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<title>bioRxiv Channel: University of California, San Francisco</title>
<link>https://biorxiv.org</link>
<description>
This feed contains articles for bioRxiv Channel "University of California, San Francisco"
</description>

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<prism:publicationName>bioRxiv</prism:publicationName>
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<title>bioRxiv</title>
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<link>https://biorxiv.org</link>
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<item rdf:about="https://biorxiv.org/content/10.1101/102145v1?rss=1">
<title>
<![CDATA[
Experimental Zika Virus Infection in a New World Monkey Model Reproduces Key Features of the Human Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/102145v1?rss=1</link>
<description><![CDATA[
Human infections by Zika virus (ZIKV), a mosquito-borne flavivirus, are associated with a current widespread outbreak in the Americas, and have been associated with neurological complications and adverse fetal outcomes such as microcephaly in pregnant women. A suitable non-human primate model is urgently needed. To evaluate ZIKV infectivity, pathogenesis, and persistence, we inoculated 4 marmosets with ZIKV and followed them by clinical monitoring and serial sampling of body fluids for up to 11 weeks. We found that marmosets experimentally infected with ZIKV reproduced key features of the human disease, including (1) asymptomatic infection, (2) brief period of detectable virus in serum (<1 week), (3) detection in other body fluids (urine, saliva, semen, and stool) for at least 2 weeks following acute infection, and (4) persistence in lymph nodes, but not other tissues, at 1 month post-infection. ZIKV-positive saliva and serum samples, but not urine, were found to be infectious in cell culture. By day 6 post-inoculation, most marmosets exhibited detectable neutralizing antibody responses concurrent with activation of NK cell and B cell subsets and an increase in circulating cytokines associated with type II interferon signaling, Transcriptome profiling revealed enrichment of immune responses to active viral infection, with up-regulation of both type I and II interferon signaling pathways, anduncovered potential host biomarkers. These results suggest that a New World monkey model of acute ZIKV infection mimics the human disease, and is likely to be useful for testing of drug and vaccine candidates.
]]></description>
<dc:creator>Chiu, C. Y.</dc:creator>
<dc:creator>Bouquet, J.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Yagi, S.</dc:creator>
<dc:creator>Sanchez San Martin, C.</dc:creator>
<dc:creator>Tamhankar, M.</dc:creator>
<dc:creator>Hodara, V. L.</dc:creator>
<dc:creator>Parodi, L. L.</dc:creator>
<dc:creator>Somasekar, S.</dc:creator>
<dc:creator>Yu, G.</dc:creator>
<dc:creator>Giavedoni, L. D.</dc:creator>
<dc:creator>Tardif, S.</dc:creator>
<dc:creator>Patterson, J.</dc:creator>
<dc:date>2017-01-21</dc:date>
<dc:identifier>doi:10.1101/102145</dc:identifier>
<dc:title><![CDATA[Experimental Zika Virus Infection in a New World Monkey Model Reproduces Key Features of the Human Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.06.475289v1?rss=1">
<title>
<![CDATA[
Autoantibody and hormone activation of the thyrotropin G protein-coupled receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.06.475289v1?rss=1</link>
<description><![CDATA[
Thyroid hormones are vital to growth and metabolism. Thyroid hormone synthesis is controlled by thyrotropin (TSH), which acts at the thyrotropin receptor (TSHR). Autoantibodies that activate the TSHR pathologically increase thyroid hormones in Graves disease. How autoantibodies mimic TSH function remains unclear. We determined cryogenic-electron microscopy structures of active and inactive TSHR. In inactive TSHR, the extracellular domain lies close to the membrane bilayer. TSH selects an upright conformation of the extracellular domain due to steric clashes between a conserved hormone glycan and the membrane bilayer. An activating autoantibody selects a similar upright conformation of the extracellular domain. Conformational changes in the extracellular domain are transduced to the seven transmembrane domain via a conserved hinge domain, a tethered peptide agonist, and a phospholipid that binds within the seven transmembrane domain. Rotation of the TSHR ECD relative to the membrane bilayer is sufficient for receptor activation, revealing a shared mechanism for other glycoprotein hormone receptors that may also extend to G protein-coupled receptors with large extracellular domains.
]]></description>
<dc:creator>Faust, B.</dc:creator>
<dc:creator>Singh, I.</dc:creator>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Hoppe, N.</dc:creator>
<dc:creator>Pinto, A. F. M.</dc:creator>
<dc:creator>Muftuoglu, Y.</dc:creator>
<dc:creator>Billesboelle, C. B.</dc:creator>
<dc:creator>Saghatelian, A.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:date>2022-01-07</dc:date>
<dc:identifier>doi:10.1101/2022.01.06.475289</dc:identifier>
<dc:title><![CDATA[Autoantibody and hormone activation of the thyrotropin G protein-coupled receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/004259v1?rss=1">
<title>
<![CDATA[
Disentangling Multidimensional Spatio-Temporal Data into their Common and Aberrant Responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/004259v1?rss=1</link>
<description><![CDATA[
With the advent of high-throughput measurement techniques, scientists and engineers are starting to grapple with massive data sets and encountering challenges with how to organize, process and extract information into meaningful structures. Multidimensional spatio-temporal biological data sets such as time series gene expression with various perturbations over different cell lines, or neural spike trains across many experimental trials, have the potential to acquire insight across multiple dimensions. For this potential to be realized, we need a suitable representation to understand the data. Since a wide range of experiments and the unknown complexity of the underlying system contribute to the heterogeneity of biological data, we propose a method based on Robust Principal Component Analysis (RPCA), which is well suited for extracting principal components when there are corrupted observations. The proposed method provides us a new representation of these data sets in terms of a common and aberrant response. This representation might help users to acquire a new insight from data.nnAuthor SummaryOne of the most exciting trends and important themes in science and engineering involves the use of high-throughput measurement data. With different dimensions, for example, various perturbations, different doses of drug or cell lines characteristics, such multidimensional data sets enable us to understand commonalities and differences across multiple dimensions. A general question is how to organize the observed data into meaningful structures and how to find an appropriate similarity measure. A natural way of viewing these complex high dimensional data sets is to examine and analyze the large-scale features and then to focus on the interesting details. With this notion, we propose an RPCA-based method which models common variations as approximately the low-rank component and anomalies as the sparse component. We show that the proposed method is able to find distinct subtypes and classify data sets in a robust way without any prior knowledge by separating these common responses and abnormal responses.
]]></description>
<dc:creator>Young Hwan Chang</dc:creator>
<dc:creator>Jim Korkola</dc:creator>
<dc:creator>Dhara N. Amin</dc:creator>
<dc:creator>Mark M. Moasser</dc:creator>
<dc:creator>Jose M. Carmena</dc:creator>
<dc:creator>Joe W. Gray</dc:creator>
<dc:creator>Claire J. Tomlin</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-04-23</dc:date>
<dc:identifier>doi:10.1101/004259</dc:identifier>
<dc:title><![CDATA[Disentangling Multidimensional Spatio-Temporal Data into their Common and Aberrant Responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/009100v1?rss=1">
<title>
<![CDATA[
Conservation and structural analysis of the Xenopus laevis phospho-proteome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/009100v1?rss=1</link>
<description><![CDATA[
The African clawed frog Xenopus laevis is an important model organism for studies in developmental and cell biology, including cell-signaling. However, our knowledge of X. laevis protein post-translational modifications remains scarce. Here, we used a mass spectrometry-based approach to survey the phosphoproteome of this species, compiling a list of 3225 phosphosites. We used this resource to study the conservation between the phosphoproteomes of X. laevis and 13 other species. We found that the degree of conservation of phosphorylation across species is predictive of sites with known molecular function, kinase interactions and functionally relevant phospho-regulatory interactions. In addition, using comparative protein structure models, we find that phosphosites within structured domains tend to be located at positions with high conformational flexibility. A fraction of sites appear to occur in inaccessible positions and have the potential to regulate protein conformation.
]]></description>
<dc:creator>Jeffrey R Johnson</dc:creator>
<dc:creator>Silvia D Santos</dc:creator>
<dc:creator>Tasha Johnson</dc:creator>
<dc:creator>Ursula Pieper</dc:creator>
<dc:creator>Andrej Sali</dc:creator>
<dc:creator>Nevan J Krogan</dc:creator>
<dc:creator>Pedro Beltrao</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-09-14</dc:date>
<dc:identifier>doi:10.1101/009100</dc:identifier>
<dc:title><![CDATA[Conservation and structural analysis of the Xenopus laevis phospho-proteome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/011569v1?rss=1">
<title>
<![CDATA[
Heterogeneous Network Edge Prediction: A Data Integration Approach to Prioritize Disease-Associated Genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/011569v1?rss=1</link>
<description><![CDATA[
The first decade of Genome Wide Association Studies (GWAS) has uncovered a wealth of disease-associated variants. Two important derivations will be the translation of this information into a multiscale understanding of pathogenic variants, and leveraging existing data to increase the power of existing and future studies through prioritization. We explore edge prediction on heterogeneous networks--graphs with multiple node and edge types--for accomplishing both tasks. First we constructed a network with 18 node types--genes, diseases, tissues, pathophysiologies, and 14 MSigDB (molecular signatures database) collections--and 19 edge types from high-throughput publicly-available resources. From this network composed of 40,343 nodes and 1,608,168 edges, we extracted features that describe the topology between specific genes and diseases. Next, we trained a model from GWAS associations and predicted the probability of association between each protein-coding gene and each of 29 well-studied complex diseases. The model, which achieved 132-fold enrichment in precision at 10% recall, outperformed any individual domain, highlighting the benefit of integrative approaches. We identified pleiotropy, transcriptional signatures of perturbations, pathways, and protein interactions as fundamental mechanisms explaining pathogenesis. Our method successfully predicted the results (with AUROC = 0.79) from a withheld multiple sclerosis (MS) GWAS despite starting with only 13 previously associated genes. Finally, we combined our network predictions with statistical evidence of association to propose four novel MS genes, three of which (JAK2, REL, RUNX3) validated on the masked GWAS. Furthermore, our predictions provide biological support highlighting REL as the causal gene within its gene-rich locus. Users can browse all predictions online (http://het.io). Heterogeneous network edge prediction effectively prioritized genetic associations and provides a powerful new approach for data integration across multiple domains.nnAuthor SummaryFor complex human diseases, identifying the genes harboring susceptibility variants has taken on medical importance. Disease-associated genes provide clues for elucidating disease etiology, predicting disease risk, and highlighting therapeutic targets. Here, we develop a method to predict whether a given gene and disease are associated. To capture the multitude of biological entities underlying pathogenesis, we constructed a heterogeneous network, containing multiple node and edge types. We built on a technique developed for social network analysis, which embraces disparate sources of data to make predictions from heterogeneous networks. Using the compendium of associations from genome-wide studies, we learned the influential mechanisms underlying pathogenesis. Our findings provide a novel perspective about the existence of pervasive pleiotropy across complex diseases. Furthermore, we suggest transcriptional signatures of perturbations are an underutilized resource amongst prioritization approaches. For multiple sclerosis, we demonstrated our ability to prioritize future studies and discover novel susceptibility genes. Researchers can use these predictions to increase the statistical power of their studies, to suggest the causal genes from a set of candidates, or to generate evidence-based experimental hypothesis.
]]></description>
<dc:creator>Daniel S Himmelstein</dc:creator>
<dc:creator>Sergio E Baranzini</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-12-11</dc:date>
<dc:identifier>doi:10.1101/011569</dc:identifier>
<dc:title><![CDATA[Heterogeneous Network Edge Prediction: A Data Integration Approach to Prioritize Disease-Associated Genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/012930v1?rss=1">
<title>
<![CDATA[
From deep TLS validation to ensembles of atomic models built from elemental motions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/012930v1?rss=1</link>
<description><![CDATA[
The widely used Translation Libration Screw (TLS) approximation describes concerted motions of atomic groups in X-ray refinement. TLS refinement often provides a better interpretation of diffraction data and the resulting rigid body motions may subsequently be assigned biochemical significance. In TLS refinement, three matrices (T, L and S) describe harmonic vibration, libration and their correlation. Because these matrices describe specific motions, they impose a number of conditions on their elements. Ignoring these conditions while refining the matrix elements may result in matrices that cannot be interpreted in terms of physically realistic motions. We describe a mathematical framework and the computational tools to analyze refined TLS matrices through their decomposition into descriptors of underlying motions. This allows for straightforward validation and identification of implausible TLS parameters. An algorithm for the generation of structural ensembles that are consistent with given TLS parameters, implemented as a part of the Phenix project, is also described.nnSynopsisProcedures for the decomposition of TLS matrices into elementary vibrations and librations indicates possible errors in the definition of these matrices and corrects them when possible. The program outputs the corresponding vibration-libration parameters and generates structural ensembles.
]]></description>
<dc:creator>Alexandre Urzhumtsev</dc:creator>
<dc:creator>Pavel Afonine</dc:creator>
<dc:creator>Andrew H Van Benschoten</dc:creator>
<dc:creator>James Fraser</dc:creator>
<dc:creator>Paul D Adams</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-01-02</dc:date>
<dc:identifier>doi:10.1101/012930</dc:identifier>
<dc:title><![CDATA[From deep TLS validation to ensembles of atomic models built from elemental motions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/012955v1?rss=1">
<title>
<![CDATA[
Predicting X-ray Diffuse Scattering from Translation Libration Screw Structural Ensembles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/012955v1?rss=1</link>
<description><![CDATA[
Identifying the intramolecular motions of proteins and nucleic acids is a major challenge in macromolecular X-ray crystallography. While Bragg diffraction describes the average positional distribution of crystalline atoms, many different models can fit this distribution equally well. Diffuse X-ray scattering can reduce this degeneracy by directly reporting on correlated atomic displacements. Although recent technological advances are increasing the potential to accurately measure diffuse scattering, computational modeling and validation tools are still needed to quantify the agreement between experimental data and different parameterizations of crystalline disorder. A new tool, phenix.diffuse, addresses this need by employing Guiniers equation to calculate diffuse scattering from Protein Data Bank (PDB)-formatted structural ensembles. As an example case, phenix.diffuse is applied to Translation-Libration-Screw (TLS) refinement, which models rigid body displacement for segments of the macromolecule. To enable calculation of diffuse scattering from TLS refined structures, phenix.tls_models builds multi-model PDB files that sample the underlying T, L and S tensors. In the glycerophosphodiesterase GpdQ, alternative TLS group partitioning and different motional correlations between groups yield markedly dissimilar diffuse scattering maps with distinct implications for molecular mechanism and allostery. These methods demonstrate how X-ray diffuse scattering can extend macromolecular structural refinement, validation, and analysis.nnSynopsisA method of simulating X-ray diffuse scattering from multi-model PDB files is presented. Despite similar agreement with Bragg data, different Translation-Libration-Screw refinement strategies produce unique diffuse intensity patterns.
]]></description>
<dc:creator>Andrew Van Benschoten</dc:creator>
<dc:creator>Pavel Afonine</dc:creator>
<dc:creator>Thomas Terwilliger</dc:creator>
<dc:creator>Michael Wall</dc:creator>
<dc:creator>Colin Jackson</dc:creator>
<dc:creator>Nicholas Sauter</dc:creator>
<dc:creator>Paul Adams</dc:creator>
<dc:creator>Alexandre Urzhumtsev</dc:creator>
<dc:creator>James Fraser</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-01-02</dc:date>
<dc:identifier>doi:10.1101/012955</dc:identifier>
<dc:title><![CDATA[Predicting X-ray Diffuse Scattering from Translation Libration Screw Structural Ensembles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/014738v1?rss=1">
<title>
<![CDATA[
Side-chain-directed model and map validation for 3D Electron Cryomicroscopy 
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</title>
<link>https://biorxiv.org/content/10.1101/014738v1?rss=1</link>
<description><![CDATA[
Advances in electron cryomicroscopy allow for the building of de novo atomic models into high resolution Coulomb potential maps. While established validation metrics independently assess map quality and model geometry, methods to assess the precise fitting of an atomic model into the map and to validate the interpretation of high resolution features are less well developed. Here, we present EMRinger, which tests model-to-map agreement using side-chain dihedral-directed map density measurements. These measurements reveal local map density peaks and show that peaks located at rotameric angles are a sensitive marker of whether the backbone is correctly positioned. The EMRinger Score can be improved by model refinement, suggesting its utility as an effective model-to-map validation metric. Additionally, EMRinger sampling identifies how radiation damage alters scattering from negatively charged amino acids during data collection. EMRinger will be useful in assessing how advances in cryo-EM increase the ability to resolve and model high-resolution features.
]]></description>
<dc:creator>Benjamin A Barad</dc:creator>
<dc:creator>Nathaniel Echols</dc:creator>
<dc:creator>Ray Yu-Ruei Wang</dc:creator>
<dc:creator>Yifan Cheng</dc:creator>
<dc:creator>Frank DiMaio</dc:creator>
<dc:creator>Paul D Adams</dc:creator>
<dc:creator>James S Fraser</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-02-03</dc:date>
<dc:identifier>doi:10.1101/014738</dc:identifier>
<dc:title><![CDATA[Side-chain-directed model and map validation for 3D Electron Cryomicroscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/014902v1?rss=1">
<title>
<![CDATA[
Coevolutionary analyses require phylogenetically deep alignments and better null models to accurately detect inter-protein contacts within and between species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/014902v1?rss=1</link>
<description><![CDATA[
When biomolecules physically interact, natural selection operates on them jointly. Contacting positions in protein and RNA structures exhibit correlated patterns of sequence evolution due to constraints imposed by the interaction, and molecular arms races can develop between interacting proteins in pathogens and their hosts. To evaluate how well methods developed to detect coevolving residues within proteins can be adapted for cross-species, inter-protein analysis, we used statistical criteria to quantify the performance of these methods in detecting inter-protein residues within 8 angstroms of each other in the co-crystal structures of 33 bacterial protein interactions. We also evaluated their performance for detecting known residues at the interface of a host-virus protein complex with a partially solved structure. Our quantitative benchmarking showed that all coevolutionary methods clearly benefit from alignments with many sequences. Methods that aim to detect direct correlations generally outperform other approaches. However, faster mutual information based methods are occasionally competitive in small alignments and with relaxed false positive rates. All commonly used null distributions are anti-conservative and have high false positive rates in some scenarios, although the empirical distribution of scores performs reasonably well with deep alignments. We conclude that coevolutionary analysis of cross-species protein interactions holds great promise but requires sequencing many more species pairs.
]]></description>
<dc:creator>Aram Avila-Herrera</dc:creator>
<dc:creator>Katherine Pollard</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-02-06</dc:date>
<dc:identifier>doi:10.1101/014902</dc:identifier>
<dc:title><![CDATA[Coevolutionary analyses require phylogenetically deep alignments and better null models to accurately detect inter-protein contacts within and between species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/015917v1?rss=1">
<title>
<![CDATA[
Selection and explosive growth may hamper the performance of rare variant association tests 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/015917v1?rss=1</link>
<description><![CDATA[
Much recent debate has focused on the role of rare variants in complex phenotypes. However, it is well known that rare alleles can only contribute a substantial proportion of the phenotypic variance when they have much larger effect sizes than common variants, which is most easily explained by natural selection constraining trait-altering alleles to low frequency. It is also plausible that demographic events will influence the genetic architecture of complex traits. Unfortunately, most rare variant association tests do not explicitly model natural selection or non-equilibrium demography. Here, we develop a novel evolutionary model of complex traits. We perform numerical calculations and simulate phenotypes under this model using inferred human demographic and selection parameters. We show that rare variants only contribute substantially to complex traits under very strong assumptions about the relationship between effect size and selection strength. We then assess the performance of state-of-the-art rare variant tests using our simulations across a broad range of model parameters. Counterintuitively, we find that statistical power is lowest when rare variants make the greatest contribution to the additive variance, and that power is substantially lower under our model than previously studied models. While many empirical studies have attempted to identify causal loci using rare variant association methods, few have reported novel associations. Some authors have interpreted this to mean that rare variants contribute little to heritability, but our results show that an alternative explanation is that rare variant tests have less power than previously estimated.
]]></description>
<dc:creator>Lawrence H. Uricchio</dc:creator>
<dc:creator>John S. Witte</dc:creator>
<dc:creator>Ryan D. Hernandez</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-03-02</dc:date>
<dc:identifier>doi:10.1101/015917</dc:identifier>
<dc:title><![CDATA[Selection and explosive growth may hamper the performance of rare variant association tests]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/016733v1?rss=1">
<title>
<![CDATA[
Mapping the Conformational Landscape of a Dynamic Enzyme by XFEL and Multitemperature Crystallography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/016733v1?rss=1</link>
<description><![CDATA[
Determining the interconverting conformations of dynamic proteins in atomic detail is a major challenge for structural biology. Conformational heterogeneity in the active site of the dynamic enzyme cyclophilin A (CypA) has been previously linked to its catalytic function, but the extent to which the different conformations of these residues are correlated is unclear. Here we compare the conformational ensembles of CypA by multitemperature synchrotron crystallography and fixed-target X-ray free electron laser (XFEL) crystallography. The "diffraction-before-destruction" nature of XFEL experiments provides a radiation-damage-free view of the functionally important alternative conformations of CypA, confirming earlier synchrotron-based results. We monitored the temperature dependences of these alternative conformations with eight synchrotron datasets spanning 100-310 K. Multiconformer models show that many alternative conformations in CypA are populated only at 240 K and above, yet others remain populated or become populated at 180 K and below. These results point to a complex evolution of conformational heterogeneity between 180-240 K that involves both thermal deactivation and solvent-driven arrest of protein motions in the crystal. The lack of a single shared conformational response to temperature within the dynamic active-site network provides evidence for a conformation shuffling model, in which exchange between rotamer states for a large aromatic ring in the middle of the network shifts the conformational ensemble for the other residues in the network. Together, our multitemperature analyses and XFEL data motivate a new generation of temperature- and time-resolved experiments to structurally characterize the dynamic underpinnings of protein function.
]]></description>
<dc:creator>Daniel A Keedy</dc:creator>
<dc:creator>Lillian R Kenner</dc:creator>
<dc:creator>Matthew Warkentin</dc:creator>
<dc:creator>Rahel Woldeyes</dc:creator>
<dc:creator>Michael C Thompson</dc:creator>
<dc:creator>Aaron S Brewster</dc:creator>
<dc:creator>Andrew H Van Benschoten</dc:creator>
<dc:creator>Elizabeth L Baxter</dc:creator>
<dc:creator>Jesse B Hopkins</dc:creator>
<dc:creator>Monarin Uervirojnangkoorn</dc:creator>
<dc:creator>Scott E McPhillps</dc:creator>
<dc:creator>Jinhu Song</dc:creator>
<dc:creator>Roberto Alonso-Mori</dc:creator>
<dc:creator>James M Holton</dc:creator>
<dc:creator>William I Weis</dc:creator>
<dc:creator>Axel T Brunger</dc:creator>
<dc:creator>S. Michael Soltis</dc:creator>
<dc:creator>Henrik Lemke</dc:creator>
<dc:creator>Ana Gonzalez</dc:creator>
<dc:creator>Nicholas K Sauter</dc:creator>
<dc:creator>Aina E Cohen</dc:creator>
<dc:creator>Henry van den Bedem</dc:creator>
<dc:creator>Robert E Thorne</dc:creator>
<dc:creator>James Fraser</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-03-19</dc:date>
<dc:identifier>doi:10.1101/016733</dc:identifier>
<dc:title><![CDATA[Mapping the Conformational Landscape of a Dynamic Enzyme by XFEL and Multitemperature Crystallography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/018879v1?rss=1">
<title>
<![CDATA[
Exposing hidden alternative backbone conformations in X-ray crystallography using qFit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/018879v1?rss=1</link>
<description><![CDATA[
Proteins must move between different conformations of their native ensemble to perform their functions. Crystal structures obtained from high-resolution X-ray diffraction data reflect this heterogeneity as a spatial and temporal conformational average. Although movement between natively populated alternative conformations can be critical for characterizing molecular mechanisms, it is challenging to identify these conformations within electron density maps. Alternative side chain conformations are generally well separated into distinct rotameric conformations, but alternative backbone conformations can overlap at several atomic positions. Our model building program qFit uses mixed integer quadratic programming (MIQP) to evaluate an extremely large number of combinations of sidechain conformers and backbone fragments to locally explain the electron density. Here, we describe two major modeling enhancements to qFit: peptide flips and alternative glycine conformations. We find that peptide flips fall into four stereotypical clusters and are enriched in glycine residues at the n+1 position. The potential for insights uncovered by new peptide flips and glycine conformations is exemplified by HIV protease, where different inhibitors are associated with peptide flips in the "flap" regions adjacent to the inhibitor binding site. Our results paint a picture of peptide flips as conformational switches, often enabled by glycine flexibility, that result in dramatic local rearrangements. Our results furthermore demonstrate the power of large-scale computational analysis to provide new insights into conformational heterogeneity. Overall, improved modeling of backbone heterogeneity with high-resolution X-ray data will connect dynamics to the structure-function relationship and help drive new design strategies for inhibitors of biomedically important systems.nnAuthor SummaryDescribing the multiple conformations of proteins is important for understanding the relationship between molecular flexibility and function. However, most methods for interpreting data from X-ray crystallography focus on building a single structure of the protein, which limits the potential for biological insights. Here we introduce an improved algorithm for using crystallographic data to model these multiple conformations that addresses two previously overlooked types of protein backbone flexibility: peptide flips and glycine movements. The method successfully models known examples of these types of multiple conformations, and also identifies new cases that were previously unrecognized but are well supported by the experimental data. For example, we discover glycine-driven peptide flips in the inhibitor-gating "flaps" of the drug target HIV protease that were not modeled in the original structures. Automatically modeling "hidden" multiple conformations of proteins using our algorithm may help drive biomedically relevant insights in structural biology pertaining to, e.g., drug discovery for HIV-1 protease and other therapeutic targets.
]]></description>
<dc:creator>Daniel A Keedy</dc:creator>
<dc:creator>James Fraser</dc:creator>
<dc:creator>Henry van den Bedem</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-05-04</dc:date>
<dc:identifier>doi:10.1101/018879</dc:identifier>
<dc:title><![CDATA[Exposing hidden alternative backbone conformations in X-ray crystallography using qFit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/020420v1?rss=1">
<title>
<![CDATA[
Rapid metagenomic identification of viral pathogens in clinical samples by real-time nanopore sequencing analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/020420v1?rss=1</link>
<description><![CDATA[
We report unbiased metagenomic detection of chikungunya virus (CHIKV), Ebola virus (EBOV), and hepatitis C virus (HCV) from four human blood samples by MinION nanopore sequencing coupled to a newly developed, web-based pipeline for real-time bioinformatics analysis on a computational server or laptop (MetaPORE). At titers ranging from 107-108 copies per milliliter, reads to EBOV from two patients with acute hemorrhagic fever and CHIKV from an asymptomatic blood donor were detected within 4 to 10 minutes of data acquisition, while lower titer HCV virus (1x105 copies per milliliter) was detected within 40 minutes. Analysis of mapped nanopore reads alone, despite an average individual error rate of 24% [range 8-49%], permitted identification of the correct viral strain in all 4 isolates, and 90% of the genome of CHIKV was recovered with >98% accuracy. Using nanopore sequencing, metagenomic detection of viral pathogens directly from clinical samples was performed within an unprecedented <6 hours sample-to-answer turnaround time and in a timeframe amenable for actionable clinical and public health diagnostics.
]]></description>
<dc:creator>Alex L Greninger</dc:creator>
<dc:creator>Samia N Naccache</dc:creator>
<dc:creator>Scot Federman</dc:creator>
<dc:creator>Guixia Yu</dc:creator>
<dc:creator>Placide Mbala</dc:creator>
<dc:creator>Vanessa Bres</dc:creator>
<dc:creator>Jerome Bouquet</dc:creator>
<dc:creator>Doug Stryke</dc:creator>
<dc:creator>Sneha Somasekar</dc:creator>
<dc:creator>Jeffrey Linnen</dc:creator>
<dc:creator>Roger Dodd</dc:creator>
<dc:creator>Prime Mulembakani</dc:creator>
<dc:creator>Bradley Schneider</dc:creator>
<dc:creator>Jean-Jacques Muyembe</dc:creator>
<dc:creator>Susan Stramer</dc:creator>
<dc:creator>Charles Y Chiu</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-06-03</dc:date>
<dc:identifier>doi:10.1101/020420</dc:identifier>
<dc:title><![CDATA[Rapid metagenomic identification of viral pathogens in clinical samples by real-time nanopore sequencing analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/024521v1?rss=1">
<title>
<![CDATA[
BRAINformat: A Data Standardization Framework for Neuroscience Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/024521v1?rss=1</link>
<description><![CDATA[
Neuroscience is entering the era of  extreme data with little experience and few plans for the associated volume, velocity, variety, and veracity challenges. This is a serious impediment for both the sharing of data across labs, as well as the utilization of modern and high-performance computing capabilities to enable data driven discovery. Here, we introduce BRAINformat, a novel file format and model for management and storage of neuroscience data. The BRAINformat library defines application-independent design concepts and modules that together create a general framework for standardization of scientific data.nnWe describe the formal specification of scientific data standards, which facilitates sharing and verification of data and formats. We introduce the concept of Managed Objects, enabling semantic components of data formats to be specified as self-contained units, supporting modular and reusable design of data format components and file storage. The BRAINformat is built off of HDF5, enabling portable, scalable, and self-describing data storage. We introduce the novel concept of Relationship Attributes for modeling and use of semantic relationships between data objects, and discuss the annotation of data using dedicated data annotation modules provided by the BRAINformat library. Based on these concepts we implement dedicated, application-oriented modules and design a data standard for neuroscience data. The BRAINformat software library is open source, easy-to-use, and provides detailed user and developer documentation and is freely available at: https://bitbucket.org/oruebel/brainformat.
]]></description>
<dc:creator>Oliver Rübel</dc:creator>
<dc:creator>Mr. Prabhat</dc:creator>
<dc:creator>Peter Denes</dc:creator>
<dc:creator>David Conant</dc:creator>
<dc:creator>Edward Chang</dc:creator>
<dc:creator>Kristofer Bouchard</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-08-13</dc:date>
<dc:identifier>doi:10.1101/024521</dc:identifier>
<dc:title><![CDATA[BRAINformat: A Data Standardization Framework for Neuroscience Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/025064v1?rss=1">
<title>
<![CDATA[
SFS_CODE: More Efficient and Flexible Forward Simulations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/025064v1?rss=1</link>
<description><![CDATA[
SUMMARYModern implementations of forward population genetic simulations are efficient and flexible, enabling the exploration of complex models that may otherwise be intractable. Here we describe an updated version of SFS_CODE, which has increased efficiency and includes many novel features. Among these features is an arbitrary model of dominance, the ability to simulate partial and soft selective sweeps, as well as track the trajectories of mutations and/or ancestries across multiple populations under complex models that are not possible under a coalescent framework. We also release sfs_coder, a Python wrapper to SFS_CODE allowing the user to easily generate command lines for common models of demography, selection, and human genome structure, as well as parse and simulate phenotypes from SFS_CODE output.nnAvailability and ImplementationOur open source software is written in C and Python, and are available under the GNU General Public License at http://sfscode.sourceforge.net.nnContactryan.hernandez@ucsf.edunnSupplementary informationDetailed usage information is available from the project website at http://sfscode.sourceforge.net.
]]></description>
<dc:creator>Ryan D. Hernandez</dc:creator>
<dc:creator>Lawrence H. Uricchio</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-08-20</dc:date>
<dc:identifier>doi:10.1101/025064</dc:identifier>
<dc:title><![CDATA[SFS_CODE: More Efficient and Flexible Forward Simulations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/025098v1?rss=1">
<title>
<![CDATA[
Statistical Inference of a Convergent Antibody Repertoire Response to Influenza Vaccine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/025098v1?rss=1</link>
<description><![CDATA[
BackgroundVaccines dramatically affect an individuals adaptive immune system, and thus provide an excellent means to study human immunity. Upon vaccination, the B cells that express antibodies (Abs) that happen to bind the vaccine are stimulated to proliferate and undergo mutagenesis at their Ab locus. This process may alter the composition of B cell lineages within an individual, which are known collectively as the antibody repertoire (AbR). Antibodies are also highly expressed in whole blood, potentially enabling unbiased RNA sequencing technologies to query this diversity. Less is known about the diversity of AbR responses across individuals to a given vaccine and if individuals tend to yield a similar response to the same antigenic stimulus.nnMethodsHere we implement a bioinformatic pipeline that extracts the AbR information from a time-series RNA-seq dataset of 5 patients who were administered a seasonal trivalent influenza vaccine (TIV). We harness the detailed time-series nature of this dataset and use methods based in functional data analysis (FDA) to identify the B cell lineages that respond to the vaccine. We then design and implement rigorous statistical tests in order to ask whether or not these patients exhibit a convergent AbR response to the same TIV.nnResultsWe find that high-resolution time-series data can be used to help identify the Ab lineages that respond to an antigenic stimulus, and that this response can exhibit a convergent nature across patients inoculated with the same vaccine. However, correlations in AbR diversity among individuals prior to inoculation can confound inference of a convergent signal unless it is taken into account.nnConclusionsWe developed a framework to identify the elements of an AbR that respond to an antigen. This information could be used to understand the diversity of different immune responses in different individuals, as well as to gauge the effectiveness of the immune response to a given stimulus within an individual. We also present a framework for testing a convergent hypothesis between AbRs; a hypothesis that is more difficult to test than previously appreciated. Our discovery of a convergent signal suggests that similar epitopes do select for antibodies with similar sequence characteristics.
]]></description>
<dc:creator>Nicolas Strauli</dc:creator>
<dc:creator>Ryan Hernandez</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-08-20</dc:date>
<dc:identifier>doi:10.1101/025098</dc:identifier>
<dc:title><![CDATA[Statistical Inference of a Convergent Antibody Repertoire Response to Influenza Vaccine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/025452v1?rss=1">
<title>
<![CDATA[
Determination of Ubiquitin Fitness Landscapes Under Different Chemical Stresses in a Classroom Setting 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/025452v1?rss=1</link>
<description><![CDATA[
Ubiquitination is an essential post-translational regulatory process that can control protein stability, localization, and activity. Ubiquitin is essential for eukaryotic life and is highly conserved, varying in only 3 amino acid positions between yeast and humans. However, recent deep sequencing studies in S. cerevisiae indicate that ubiquitin is highly tolerant to single amino acid mutations. To resolve this paradox, we hypothesized that the set of tolerated substitutions would be reduced when the cultures are not grown in rich media conditions and that chemically induced physiologic perturbations might unmask constraints on the ubiquitin sequence. To test this hypothesis, a class of first year UCSF graduate students employed a deep mutational scanning procedure to determine the fitness landscape of a library of all possible single amino acid mutations of ubiquitin in the presence of one of five small molecule perturbations: MG132, Dithiothreitol (DTT), Hydroxyurea (HU), Caffeine, and DMSO. Our data reveal that the number of tolerated substitutions is greatly reduced by DTT, HU, or Caffeine, and that these perturbations uncover "shared sensitized positions" localized to areas around the hydrophobic patch and to the C-terminus. We also show perturbation specific effects including the sensitization of His68 in HU and tolerance to mutation at Lys63 in DTT. Taken together, our data suggest that chemical stress reduces buffering effects in the ubiquitin proteasome system, revealing previously hidden fitness defects. By expanding the set of chemical perturbations assayed, potentially by other classroom-based experiences, we will be able to further address the apparent dichotomy between the extreme sequence conservation and the experimentally observed mutational tolerance of ubiquitin. Finally, this study demonstrates the realized potential of a project lab-based interdisciplinary graduate curriculum.
]]></description>
<dc:creator>David Mavor</dc:creator>
<dc:creator>Participants in UCSF PUBS Class</dc:creator>
<dc:creator>James Fraser</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-08-25</dc:date>
<dc:identifier>doi:10.1101/025452</dc:identifier>
<dc:title><![CDATA[Determination of Ubiquitin Fitness Landscapes Under Different Chemical Stresses in a Classroom Setting]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/027631v1?rss=1">
<title>
<![CDATA[
Tyrosination of α-Tubulin Controls The Initiation of Processive Dynein-Dynactin Motility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/027631v1?rss=1</link>
<description><![CDATA[
Post-translational modifications (PTMs) of /{beta}-tubulin are believed to regulate interactions with microtubule binding proteins. A well-characterized PTM involves the removal and re-ligation of the C-terminal tyrosine on -tubulin, but the purpose of this tyrosination-detyrosination cycle remains elusive. Here, we examined the processive motility of mammalian dynein complexed with dynactin and BicD2 (DDB) on tyrosinated versus detyrosinated microtubules. Motility was decreased ~4-fold on detyrosinated microtubules, constituting the largest effect of a tubulin PTM on motor function observed to date. This preference is mediated by dynactins microtubule binding p150 subunit rather than dynein itself. Interestingly, on chimeric microtubules, DDB molecules that initiated movement on tyrosinated tubulin continued moving into a region of detyrosinated tubulin. This result indicates that the -tubulin tyrosine facilitates initial motor-tubulin encounters, but is not needed for subsequent motility. Our results reveal a strong effect of the C-terminal -tubulin tyrosine on dynein-dynactin motility and suggest that the tubulin tyrosination cycle could modulate the initiation of dynein-driven motility in cells.
]]></description>
<dc:creator>Richard J McKenney</dc:creator>
<dc:creator>Walter Huynh</dc:creator>
<dc:creator>Ronald D. Vale</dc:creator>
<dc:creator>Minhaj Sirajuddin</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-09-25</dc:date>
<dc:identifier>doi:10.1101/027631</dc:identifier>
<dc:title><![CDATA[Tyrosination of α-Tubulin Controls The Initiation of Processive Dynein-Dynactin Motility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/033746v1?rss=1">
<title>
<![CDATA[
Measuring and modeling diffuse scattering in protein X-ray crystallography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/033746v1?rss=1</link>
<description><![CDATA[
X-ray diffraction has the potential to provide rich information about the structural dynamics of macromolecules. To realize this potential, both Bragg scattering, which is currently used to derive macromolecular structures, and diffuse scattering, which reports on correlations in charge density variations must be measured. Until now measurement of diffuse scattering from protein crystals has been scarce, due to the extra effort of collecting diffuse data. Here, we present three-dimensional measurements of diffuse intensity collected from crystals of the enzymes cyclophilin A and trypsin. The measurements were obtained from the same X-ray diffraction images as the Bragg data, using best practices for standard data collection. To model the underlying dynamics in a practical way that could be used during structure refinement, we tested Translation-Libration-Screw (TLS), Liquid-Like Motions (LLM), and coarse-grained Normal Modes (NM) models of protein motions. The LLM model provides a global picture of motions and were refined against the diffuse data, while the TLS and NM models provide more detailed and distinct descriptions of atom displacements, and only used information from the Bragg data. Whereas different TLS groupings yielded similar Bragg intensities, they yielded different diffuse intensities, none of which agreed well with the data. In contrast, both the LLM and NM models agreed substantially with the diffuse data. These results demonstrate a realistic path to increase the number of diffuse datasets available to the wider biosciences community and indicate that NM-based refinement can generate dynamics-inspired structural models that simultaneously agree with both Bragg and diffuse scattering.nnSignificanceThe structural details of protein motions are critical to understanding many biological processes, but they are often hidden to conventional biophysical techniques. Diffuse X-ray scattering can reveal details of the correlated movements between atoms; however, the data collection historically has required extra effort and dedicated experimental protocols. We have measured three-dimensional diffuse intensities in X-ray diffraction from CypA and trypsin crystals using standard crystallographic data collection techniques. Analysis of the resulting data is consistent with the protein motions resembling diffusion in a liquid or vibrations of a soft solid. Our results show that using diffuse scattering to model protein motions can become a component of routine crystallographic analysis through the extension of commonplace methods.
]]></description>
<dc:creator>Andrew H Van Benschoten</dc:creator>
<dc:creator>Lin Liu</dc:creator>
<dc:creator>Ana Gonzalez</dc:creator>
<dc:creator>Aaron S Brewster</dc:creator>
<dc:creator>Nicholas K Sauter</dc:creator>
<dc:creator>James S Fraser</dc:creator>
<dc:creator>Michael Wall</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-12-06</dc:date>
<dc:identifier>doi:10.1101/033746</dc:identifier>
<dc:title><![CDATA[Measuring and modeling diffuse scattering in protein X-ray crystallography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/034017v1?rss=1">
<title>
<![CDATA[
Bat Accelerated Regions Identify a Bat Forelimb Specific Enhancer in the HoxD Locus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/034017v1?rss=1</link>
<description><![CDATA[
The molecular events leading to the development of the bat wing remain largely unknown, and are thought to be caused, in part, by changes in gene expression during limb development. These expression changes could be instigated by variations in gene regulatory enhancers. Here, we used a comparative genomics approach to identify regions that evolved rapidly in the bat ancestor but are highly conserved in other vertebrates. We discovered 166 bat accelerated regions (BARs) that overlap H3K27ac and p300 ChIP-seq peaks in developing mouse limbs. Using a mouse enhancer assay, we show that five Myotis lucifugus BARs drive gene expression in the developing mouse limb, with the majority showing differential enhancer activity compared to the mouse orthologous BAR sequences. These include BAR116, which is located telomeric to the HoxD cluster and had robust forelimb expression for the M. lucifugus sequence and no activity for the mouse sequence at embryonic day 12.5. Developing limb expression analysis of Hoxd10-Hoxd13 in Miniopterus natalensis bats showed a high-forelimb weak-hindlimb expression for Hoxd10-Hoxd11, similar to the expression trend observed for M. lucifugus BAR116 in mice, suggesting that it could be involved in the regulation of the bat HoxD complex. Combined, our results highlight novel regulatory regions that could be instrumental for the morphological differences leading to the development of the bat wing.nnAuthor SummaryThe limb is a classic example of vertebrate homology and is represented by a large range of morphological structures such as fins, legs and wings. The evolution of these structures could be driven by alterations in gene regulatory elements that have critical roles during development. To identify elements that may contribute to bat wing development, we characterized sequences that are conserved between vertebrates, but changed significantly in the bat lineage. We then overlapped these sequences with predicted developing limb enhancers as determined by ChIP-seq, finding 166 bat accelerated sequences (BARs). Five BARs that were tested for enhancer activity in mice all drove expression in the limb. Testing the mouse orthologous sequence showed that three had differences in their limb enhancer activity as compared to the bat sequence. Of these, BAR116 was of particular interest as it is located near the HoxD locus, an essential gene complex required for proper spatiotemporal patterning of the developing limb. The bat BAR116 sequence drove robust forelimb expression but the mouse BAR116 sequence did not show enhancer activity. These experiments correspond to analyses of HoxD gene expressions in developing bat limbs, which had strong forelimb versus weak hindlimb expression for Hoxd10-11. Combined, our studies highlight specific genomic regions that could be important in shaping the morphological differences that led to the development of the bat wing.
]]></description>
<dc:creator>Betty M Booker</dc:creator>
<dc:creator>Tara Friedrich</dc:creator>
<dc:creator>Mandy K Mason</dc:creator>
<dc:creator>Julia E VanderMeer</dc:creator>
<dc:creator>Jingjing Zhao</dc:creator>
<dc:creator>Walter L Eckalbar</dc:creator>
<dc:creator>Malcolm Logan</dc:creator>
<dc:creator>Nicola Illing</dc:creator>
<dc:creator>Katherine S Pollard</dc:creator>
<dc:creator>Nadav Ahituv</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-12-09</dc:date>
<dc:identifier>doi:10.1101/034017</dc:identifier>
<dc:title><![CDATA[Bat Accelerated Regions Identify a Bat Forelimb Specific Enhancer in the HoxD Locus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/035253v1?rss=1">
<title>
<![CDATA[
High-accuracy HLA type inference from whole-genome sequencing data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/035253v1?rss=1</link>
<description><![CDATA[
Extensive hyperpolymorphism and sequence similarity between the HLA genes make HLA type inference from whole-genome sequencing data a challenging problem. We address these by representing sequences from over 10,000 known alleles in a reference graph structure, enabling accurate read mapping. HLA*PRG, our algorithm, outperforms existing methods by a wide margin and for the first time consistently achieves the accuracy of gold-standard reference methods with one error across 158 alleles tested.
]]></description>
<dc:creator>Alexander Dilthey</dc:creator>
<dc:creator>Pierre-Antoine Gourraud</dc:creator>
<dc:creator>Zamin Iqbal</dc:creator>
<dc:creator>Gil McVean</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-12-24</dc:date>
<dc:identifier>doi:10.1101/035253</dc:identifier>
<dc:title><![CDATA[High-accuracy HLA type inference from whole-genome sequencing data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/036822v1?rss=1">
<title>
<![CDATA[
Differential methylation between ethnic sub-groups reflects the effect of genetic ancestry and environmental exposures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/036822v1?rss=1</link>
<description><![CDATA[
In clinical practice and biomedical research populations are often divided categorically into distinct racial/ethnic groups. In reality, these categories, which are based on social rather than biological constructs, comprise diverse groups with highly heterogeneous histories, cultures, traditions, religions, social and environmental exposures and ancestral backgrounds. Their use is thus widely debated and genetic ancestry has been suggested as a complement or alternative to this categorization. However, few studies have examined the relative contributions of racial/ethnic identity, genetic ancestry, and environmental exposures on well-established and fundamental biological processes. We examined the associations between ethnicity, ancestry, and environmental exposures and DNA methylation. We typed over 450,000 CpG sites in primary whole blood of 573 individuals of diverse Hispanic descent who also had high-density genotype data. We found that both self-identified ethnicity and genetically determined ancestry were significantly associated with methylation levels at a large number of CpG sites (916 and 194, respectively). Among loci differentially methylated between ethnic groups, a median of 75.7% (IQR 45.8% to 92%) of the variance in methylation associated with ethnicity could be accounted for by shared genomic ancestry accounts. We also found significant enrichment (p = 4.2 x 10-64) of ethnicity-associated sites amongst loci previously associated with environmental and social exposures, particularly maternal smoking during pregnancy. Our study suggests that although differential methylation between ethnic groups can be partially explained by the shared genetic ancestry, a significant effect of ethnicity is likely due to environmental, social, or cultural factors, which differ between ethnic groups.nnOne Sentence SummaryIn order to better understand the role of ethnic self-identification and genetically determined ancestry in biomedical outcomes, we explore their relative contributions to variation in methylation, a fundamental biological process.nnSources of FundingThis research was supported in part by the Sandler Family Foundation, the American Asthma Foundation, National Institutes of Health (P60 MD006902, R01 HL117004, R21ES24844, U54MD009523, R01 ES015794, R01 HL088133, M01 RR000083, R01 HL078885, R01 HL104608, U19 AI077439, M01 RR00188, U01 HG009080, and R01 HL135156), ARRA grant RC2 HL101651, and TRDRP 24RT-0025; EGB was supported in part through grants from the Flight Attendant Medical Research Institute (FAMRI), and NIH (K23 HL004464); NZ was supported in part by an NIH career development award from the NHLBI (K25HL121295). JMG was supported in part by NIH Training Grant T32 (T32GM007546) and career development awards from the NHLBI (K23HL111636) and NCATS (KL2TR000143) as well as the Hewett Fellowship; N.T. was supported in part by an institutional training grant from the NIGMS (T32-GM007546) and career development awards from the NHLBI (K12-HL119997 and K23-HL125551), Parker B. Francis Fellowship Program, and the American Thoracic Society; CRG was supported in part by NIH Training Grant T32 (GM007175) and the UCSF Chancellors Research Fellowship and Dissertation Year Fellowship; RK was supported with a career development award from the NHLBI (K23HL093023); HJF was supported in part by the GCRC (RR00188); PCA was supported in part by the Ernest S. Bazley Grant; MAS was supported in part by 1R01HL128439-01. This publication was supported by various institutes within the National Institutes of Health. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH.
]]></description>
<dc:creator>Joshua M Galanter</dc:creator>
<dc:creator>Christopher R Gignoux</dc:creator>
<dc:creator>Sam S Oh</dc:creator>
<dc:creator>Dara Torgerson</dc:creator>
<dc:creator>Maria Pino-Yanes</dc:creator>
<dc:creator>Neeta Thakur</dc:creator>
<dc:creator>Celeste Eng</dc:creator>
<dc:creator>Donglei Hu</dc:creator>
<dc:creator>Scott Huntsmann</dc:creator>
<dc:creator>Harold J Farber</dc:creator>
<dc:creator>Pedro Avila</dc:creator>
<dc:creator>Emerita Brigino-Buenaventura</dc:creator>
<dc:creator>Michael LeNoir</dc:creator>
<dc:creator>Kelly Meade</dc:creator>
<dc:creator>Denise Serebrisky</dc:creator>
<dc:creator>William Rodriguez-Cintron</dc:creator>
<dc:creator>Rajessh Kumar</dc:creator>
<dc:creator>Jose R Rodriguez-Santana</dc:creator>
<dc:creator>Max Seibold</dc:creator>
<dc:creator>Luisa Borrell</dc:creator>
<dc:creator>Esteban G Burchard</dc:creator>
<dc:creator>Noah Zaitlen</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-01-15</dc:date>
<dc:identifier>doi:10.1101/036822</dc:identifier>
<dc:title><![CDATA[Differential methylation between ethnic sub-groups reflects the effect of genetic ancestry and environmental exposures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/038117v1?rss=1">
<title>
<![CDATA[
Micro-Magellan: A flexible, open source acquisition software for high throughput biological light microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/038117v1?rss=1</link>
<description><![CDATA[
We demonstrate the capabilities of Magellan: a flexible, open source microscopy software for reproducible high throughput imaging of biological samples across heterogeneous scales of space and time. Magellan provides a simple user interface for exploration and automated imaging of non-cuboidal regions. By utilizing the hardware abstraction layer of Magellan, Magellan provides a powerful and extensible platform for imaging heterogeneous biological samples on a wide range of existing microscopes.
]]></description>
<dc:creator>Henry Pinkard</dc:creator>
<dc:creator>Nico Stuurman</dc:creator>
<dc:creator>Kaitlin Corbin</dc:creator>
<dc:creator>Ronald Vale</dc:creator>
<dc:creator>Matthew F Krummel</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-01-28</dc:date>
<dc:identifier>doi:10.1101/038117</dc:identifier>
<dc:title><![CDATA[Micro-Magellan: A flexible, open source acquisition software for high throughput biological light microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/040089v1?rss=1">
<title>
<![CDATA[
Local joint testing improves power and identifies missing heritability in association studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/040089v1?rss=1</link>
<description><![CDATA[
There is mounting evidence that complex human phenotypes are highly polygenic, with many loci harboring multiple causal variants, yet most genetic association studies examine each SNP in isolation. While this has lead to the discovery of thousands of disease associations, discovered variants account for only a small fraction of disease heritability. Alternative multi-SNP methods have been proposed, but issues such as multiple testing correction, sensitivity to genotyping error, and optimization for the underlying genetic architectures remain. Here we describe a local joint testing procedure, complete with multiple testing correction, that leverages a genetic phenomenon we call linkage masking wherein linkage disequilibrium between SNPs hides their signal under standard association methods. We show that local joint testing on the original Wellcome Trust Case Control Consortium dataset leads to the discovery of 29% more associated loci that were later found in followup studies containing thousands of additional individuals. These loci double the heritability explained by genome-wide significant associations in the WTCCC dataset, implicating linkage masking as a novel source of missing heritability. Furthermore, we show that local joint testing in a cis-eQTL study of the gEUVADIS dataset increases the number of genes discovered by 10.7% over marginal analyses. Our multiple hypothesis correction and joint testing framework are available in a python software package called jester, available at github.com/brielin/Jester.
]]></description>
<dc:creator>Brielin C Brown</dc:creator>
<dc:creator>Alkes Price</dc:creator>
<dc:creator>Nikolaos Patsopoulos</dc:creator>
<dc:creator>Noah Zaitlen</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-02-18</dc:date>
<dc:identifier>doi:10.1101/040089</dc:identifier>
<dc:title><![CDATA[Local joint testing improves power and identifies missing heritability in association studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/048181v1?rss=1">
<title>
<![CDATA[
Subtle stratification confounds estimates of heritability from rare variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/048181v1?rss=1</link>
<description><![CDATA[
Genome-wide significant associations generally explain only a small proportion of the narrow-sense heritability of complex disease (h2). While considerably more heritability is explained by all genotyped SNPs (hg2), for most traits, much heritability remains missing (hg2 < h2). Rare variants, poorly tagged by genotyped SNPs, are a major potential source of the gap between hg2 and h2. Recent efforts to assess the contribution of both sequenced and imputed rare variants to phenotypes suggest that substantial heritability may lie in these variants. Here we analyze sequenced SNPs, imputed SNPs and haploSNPs-- haplotype variants constructed from within a sample, without using a reference panel-- and show that studies of heritability from these variants may be strongly confounded by subtle population stratification. For example, when meta-analyzing heritability estimates from 22 randomly ascertained case-control traits from the GERA cohort, we observe a statistically significant increase in heritability explained by imputed SNPs even after correcting for principal components (PCs) from genotyped (or imputed) SNPs. However, this increase is eliminated when correcting for stratification using PCs from a larger number of haploSNPs. We note that subtle stratification may also impact estimates of heritability from array SNPs, although we find that this is generally a less severe problem. Overall, our results suggest that estimating the heritability explained by rare variants for case-control traits requires exquisite control for population stratification, but current methods may not provide this level of control.
]]></description>
<dc:creator>Gaurav Bhatia</dc:creator>
<dc:creator>Alexander Gusev</dc:creator>
<dc:creator>Po-Ru Loh</dc:creator>
<dc:creator>Hilary Kiyo Finucane</dc:creator>
<dc:creator>Bjarni J Vilhjalmsson</dc:creator>
<dc:creator>Stephan Ripke</dc:creator>
<dc:creator>SCZ Working Group of the Psychiatric Genomics Cons</dc:creator>
<dc:creator>Shaun Purcell</dc:creator>
<dc:creator>Eli Stahl</dc:creator>
<dc:creator>Mark Daly</dc:creator>
<dc:creator>Teresa R de Candia</dc:creator>
<dc:creator>Sang Hong Lee</dc:creator>
<dc:creator>Benjamin M Neale</dc:creator>
<dc:creator>Matthew C Keller</dc:creator>
<dc:creator>Noah A Zaitlen</dc:creator>
<dc:creator>Bogdan Pasaniuc</dc:creator>
<dc:creator>Nick Patterson</dc:creator>
<dc:creator>Jian Yang</dc:creator>
<dc:creator>Alkes L Price</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-04-13</dc:date>
<dc:identifier>doi:10.1101/048181</dc:identifier>
<dc:title><![CDATA[Subtle stratification confounds estimates of heritability from rare variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/049791v1?rss=1">
<title>
<![CDATA[
Cytokine and Leukocyte Profiling Reveal Pro-Inflammatory and Autoimmune Features in Frontotemporal Dementia Patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/049791v1?rss=1</link>
<description><![CDATA[
The growing link between systemic environment and brain function opens the possibility that cellular communication and composition in blood are correlated with brain health. We tested this concept in frontotemporal dementia with novel, unbiased tools that measure hundreds of soluble signaling proteins or characterize the vast immune cell repertoire in blood. With these tools we discovered complementary abnormalities indicative of abnormal T cell populations and autoimmunity in frontotemporal dementia.
]]></description>
<dc:creator>Philipp A. Jaeger</dc:creator>
<dc:creator>Trisha M. Stan</dc:creator>
<dc:creator>Eva Czirr</dc:creator>
<dc:creator>Markus Britschgi</dc:creator>
<dc:creator>Daniela Berdnik</dc:creator>
<dc:creator>Ruo-Pan Huang</dc:creator>
<dc:creator>Bradley F. Boeve</dc:creator>
<dc:creator>Adam L. Boxer</dc:creator>
<dc:creator>NiCole Finch</dc:creator>
<dc:creator>Gabriela K. Fragiadakis</dc:creator>
<dc:creator>Neill R. Graff-Radford</dc:creator>
<dc:creator>Ruochun Huang</dc:creator>
<dc:creator>Hudson Johns</dc:creator>
<dc:creator>Anna Karydas</dc:creator>
<dc:creator>David S. Knopman</dc:creator>
<dc:creator>Michael Leipold</dc:creator>
<dc:creator>Holde T. Maecker</dc:creator>
<dc:creator>Zachary Miller</dc:creator>
<dc:creator>Ronald Petersen</dc:creator>
<dc:creator>Rosa Rademakers</dc:creator>
<dc:creator>Chung-Huan Sun</dc:creator>
<dc:creator>Steve Younkin</dc:creator>
<dc:creator>Bruce L. Miller</dc:creator>
<dc:creator>Tony Wyss-Coray</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-04-22</dc:date>
<dc:identifier>doi:10.1101/049791</dc:identifier>
<dc:title><![CDATA[Cytokine and Leukocyte Profiling Reveal Pro-Inflammatory and Autoimmune Features in Frontotemporal Dementia Patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/051821v1?rss=1">
<title>
<![CDATA[
Chytrid fungi construct actin-rich pseudopods, implicating actin regulators WASP and SCAR in an ancient mode of cell motility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/051821v1?rss=1</link>
<description><![CDATA[
Diverse eukaryotic cells crawl through complex environments using distinct modes of migration. To understand the underlying mechanisms and their evolutionary relationships, we must define each mode, and identify its phenotypic and molecular markers. Here, we focus on a widely dispersed migration mode characterized by dynamic, actin-filled pseudopods that we call "-motility." Mining genomic data reveals a clear trend: only organisms with both WASP and SCAR/WAVE--activators of branched actin assembly--make actin-filled pseudopods. While SCAR has been shown to drive pseudopod formation, WASPs role in this process is controversial. We hypothesize that these genes together represent a genetic signature of -motility, because both are used for pseudopod formation. WASP depletion from human neutrophils confirms that both proteins are involved in explosive actin polymerization, pseudopod formation, and cell migration, and colocalize to dynamic signaling structures. Moreover, retention of WASP together with SCAR correctly predicts -motility in disease-causing chytrid fungi, which we show crawl at >30 m/min with actin-filled pseudopods. By focusing on one migration mode in many eukaryotes, we identify a genetic marker of pseudopod formation, the morphological feature of -motility, providing evidence for a widely distributed mode of cell crawling with a single evolutionary origin.
]]></description>
<dc:creator>Lillian K. Fritz-Laylin</dc:creator>
<dc:creator>Samuel J. Lord</dc:creator>
<dc:creator>R. Dyche Mullins</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-05-04</dc:date>
<dc:identifier>doi:10.1101/051821</dc:identifier>
<dc:title><![CDATA[Chytrid fungi construct actin-rich pseudopods, implicating actin regulators WASP and SCAR in an ancient mode of cell motility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/053041v1?rss=1">
<title>
<![CDATA[
Dumpster diving in RNA-sequencing to find the source of every last read 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/053041v1?rss=1</link>
<description><![CDATA[
High throughput RNA sequencing technologies have provided invaluable research opportunities across distinct scientific domains by producing quantitative readouts of the transcriptional activity of both entire cellular populations and single cells. The majority of RNA-Seq analyses begin by mapping each experimentally produced sequence (i.e., read) to a set of annotated reference sequences for the organism of interest. For both biological and technical reasons, a significant fraction of reads remains unmapped. In this work, we develop Read Origin Protocol (ROP) to discover the source of all reads originating from complex RNA molecules, recombinant T and B cell receptors, and microbial communities. We applied ROP to 8,641 samples across 630 individuals from 54 tissues. A fraction of RNA-Seq data (n=86) was obtained in-house; the remaining data was obtained from the Genotype-Tissue Expression (GTEx v6) project. To generalize the reported number of accounted reads, we also performed ROP analysis on thousands of different, randomly selected, and publicly available RNA-Seq samples in the Sequence Read Archive (SRA). Our approach can account for 99.9% of 1 trillion reads of various read length across the merged dataset (n=10641). Using in-house RNA-Seq data, we show that immune profiles of asthmatic individuals are significantly different from the profiles of control individuals, with decreased average per sample T and B cell receptor diversity. We also show that immune diversity is inversely correlated with microbial load. Our results demonstrate the potential of ROP to exploit unmapped reads in order to better understand the functional mechanisms underlying connections between the immune system, microbiome, human gene expression, and disease etiology. ROP is freely available at https://github.com/smangul1/rop and currently supports human and mouse RNA-Seq reads.
]]></description>
<dc:creator>Serghei Mangul</dc:creator>
<dc:creator>Harry Taegyun Yang</dc:creator>
<dc:creator>Nicolas Strauli</dc:creator>
<dc:creator>Franziska Gruhl</dc:creator>
<dc:creator>Timothy Daley</dc:creator>
<dc:creator>Stephanie Christenson</dc:creator>
<dc:creator>Agata Wesolowska Andersen</dc:creator>
<dc:creator>Roberto Spreafico</dc:creator>
<dc:creator>Cydney Rios</dc:creator>
<dc:creator>Celeste Eng</dc:creator>
<dc:creator>Andrew D. Smith</dc:creator>
<dc:creator>Ryan D. Hernandez</dc:creator>
<dc:creator>Roel A. Ophoff</dc:creator>
<dc:creator>Jose Rodriguez Santana</dc:creator>
<dc:creator>Prescott G. Woodruff</dc:creator>
<dc:creator>Esteban Burchard</dc:creator>
<dc:creator>Max A. Seibold</dc:creator>
<dc:creator>Sagiv Shifman</dc:creator>
<dc:creator>Eleazar Eskin</dc:creator>
<dc:creator>Noah Zaitlen</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-05-13</dc:date>
<dc:identifier>doi:10.1101/053041</dc:identifier>
<dc:title><![CDATA[Dumpster diving in RNA-sequencing to find the source of every last read]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/055285v1?rss=1">
<title>
<![CDATA[
A scalable strategy for high-throughput GFP tagging of endogenous human proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/055285v1?rss=1</link>
<description><![CDATA[
A central challenge of the post-genomic era is to comprehensively characterize the cellular role of the [~]20,000 proteins encoded in the human genome. To systematically study protein function in a native cellular background, libraries of human cell lines expressing proteins tagged with a functional sequence at their endogenous loci would be very valuable. Here, using electroporation of Cas9/sgRNA ribonucleoproteins and taking advantage of a split-GFP system, we describe a scalable method for the robust, scarless and specific tagging of endogenous human genes with GFP. Our approach requires no molecular cloning and allows a large number of cell lines to be processed in parallel. We demonstrate the scalability of our method by targeting 48 human genes and show that the resulting GFP fluorescence correlates with protein expression levels. We next present how our protocols can be easily adapted for the tagging of a given target with GFP repeats, critically enabling the study of low-abundance proteins. Finally, we show that our GFP tagging approach allows the biochemical isolation of native protein complexes for proteomic studies. Together, our results pave the way for the large-scale generation of endogenously tagged human cell lines for the proteome-wide analysis of protein localization and interaction networks in a native cellular context.nnSIGNIFICANCE STATEMENTThe function of a large fraction of the human proteome still remains poorly characterized. Tagging proteins with a functional sequence is a powerful way to access function, and inserting tags at endogenous genomic loci allows the preservation of a near-native cellular background. To characterize the cellular role of human proteins in a systematic manner and in a native context, we developed a method for tagging endogenous human proteins with GFP that is both rapid and readily applicable at a genome-wide scale. Our approach allows studying both localization and interaction partners of the protein target. Our results pave the way for the large-scale generation of endogenously tagged human cell lines for a systematic functional interrogation of the human proteome.
]]></description>
<dc:creator>Manuel Leonetti</dc:creator>
<dc:creator>Sayaka Sekine</dc:creator>
<dc:creator>Daichi Kamiyama</dc:creator>
<dc:creator>Jonathan S Weissman</dc:creator>
<dc:creator>Bo Huang</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-05-24</dc:date>
<dc:identifier>doi:10.1101/055285</dc:identifier>
<dc:title><![CDATA[A scalable strategy for high-throughput GFP tagging of endogenous human proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/056168v1?rss=1">
<title>
<![CDATA[
The Effects of Migration and Assortative Mating on Admixture Linkage Disequilibrium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/056168v1?rss=1</link>
<description><![CDATA[
1Statistical models in medical and population genetics typically assume that individuals assort randomly in a population. While this simplifies model complexity, it contradicts an increasing body of evidence of non-random mating in human populations. Specifically, it has been shown that assortative mating is significantly affected by genomic ancestry. In this work we examine the effects of ancestry-assortative mating on the linkage disequilibrium between local ancestry tracks of individuals in an admixed population. To accomplish this, we develop an extension to the Wright-Fisher model that allows for ancestry based assortative mating. We show that ancestry-assortment perturbs the distribution of local ancestry linkage disequilibrium (LAD) and the variance of ancestry in a population as a function of the number of generations since admixture. This assortment effect can induce errors in demographic inference of admixed populations when methods assume random mating. We derive closed form formulae for LAD under an assortative-mating model with and without migration. We observe that LAD depends on the correlation of global ancestry of couples in each generation, the migration rate of each of the ancestral populations, the initial proportions of ancestral populations, and the number of generations since admixture. We also present the first evidence of ancestry-assortment in African Americans and examine LAD in simulated and real admixed population data of African Americans. We find that demographic inference under the assumption of random mating significantly underestimates the number of generations since admixture, and that accounting for assortative mating using the patterns of LAD results in estimates that more closely agrees with the historical narrative.
]]></description>
<dc:creator>Noah Zaitlen</dc:creator>
<dc:creator>Scott Huntsman</dc:creator>
<dc:creator>Donglei Hu</dc:creator>
<dc:creator>Melissa Spear</dc:creator>
<dc:creator>Celeste Eng</dc:creator>
<dc:creator>Sam S Oh</dc:creator>
<dc:creator>Marquitta J White</dc:creator>
<dc:creator>Angel Mak</dc:creator>
<dc:creator>Adam Davis</dc:creator>
<dc:creator>Kelly Meade</dc:creator>
<dc:creator>Emerita Brigino-Buenaventura</dc:creator>
<dc:creator>Michael A LeNoir</dc:creator>
<dc:creator>Kirsten Bibbins-Domingo</dc:creator>
<dc:creator>Esteban G Burchard</dc:creator>
<dc:creator>Eran Halperin</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-05-31</dc:date>
<dc:identifier>doi:10.1101/056168</dc:identifier>
<dc:title><![CDATA[The Effects of Migration and Assortative Mating on Admixture Linkage Disequilibrium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/056614v1?rss=1">
<title>
<![CDATA[
Proteobacteria drive significant functional variability in the human gut microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/056614v1?rss=1</link>
<description><![CDATA[
While human gut microbiomes vary significantly in taxonomic composition, biological pathway abundance is surprisingly invariable across hosts. We hypothesized that healthy microbiomes appear functionally redundant due to factors that obscure differences in gene abundance across hosts. To account for these biases, we developed a powerful test of gene variability, applicable to shotgun metagenomes from any environment. Our analysis of healthy stool metagenomes reveals thousands of genes whose abundance differs signifi-cantly between people consistently across studies, including glycolytic enzymes, lipopolysac-charide biosynthetic genes, and secretion systems. Even housekeeping pathways contain a mix of variable and invariable genes, though most deeply conserved genes are significantly invariable. Variable genes tend to be associated with Proteobacteria, as opposed to taxa used to define enterotypes or the dominant phyla Bacteroidetes and Firmicutes. These re-sults establish limits on functional redundancy and predict specific genes and taxa that may drive physiological differences between gut microbiomes.nnImpact StatementA statistical test for gene variability reveals extensive functional differences between healthy humanmicrobiomes.
]]></description>
<dc:creator>Patrick H Bradley</dc:creator>
<dc:creator>Katherine S Pollard</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-06-02</dc:date>
<dc:identifier>doi:10.1101/056614</dc:identifier>
<dc:title><![CDATA[Proteobacteria drive significant functional variability in the human gut microbiome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/060145v1?rss=1">
<title>
<![CDATA[
Age-related differences in self-reported sleep quality predict healthy ageing across multiple domains: a multi-modal cohort of 2406 adults 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/060145v1?rss=1</link>
<description><![CDATA[
ObjectivesTo examine age related differences in self-reported sleep quality and their associations with health outcomes across four domains: Physical Health, Cognitive Health, Mental Health and Neural Health.nnSettingCam-CAN is a cohort study in East Anglia/England, which collected self-reported health and lifestyle questions as well as a range of objective measures from healthy adults.nnParticipants2406 healthy adults (age 18-98) answered questions about their sleep quality (Pittsburgh Sleep Quality Index) and measures of Physical, Cognitive, Mental, and Neural Health. A subset of 641 individuals provided measures of brain structure.nnMain outcome measuresPittsburgh Sleep Quality Index scores (PSQI) of sleep, and scores across tests within the four domains of health. Latent Class Analysis (LCA) is used to identify sleep types across the lifespan. Bayesian regressions quantify the presence, and absence, of relationships between sleep quality and health measures.nnResultsBetter sleep is generally associated with better health outcomes, strongly so for mental health, moderately for cognitive and physical health, but not for sleep quality and neural health. Latent Class Analysis identified four sleep types:  Good sleepers (68.6%, most frequent in middle age),  inefficient sleepers (13.05%, most frequent in old age),  Delayed sleepers (9.76%, most frequent in young adults) and  poor sleepers (8.6%, most frequent in old age). There is little evidence for interactions between sleep quality and age on health outcomes. Finally, we observe u-shaped associations between sleep duration and mental health (depression and anxiety) as well as self-reported general health, such that both short and long sleep were associated with poorer outcomes.nnConclusionsLifespan changes in sleep quality are multifaceted and not captured well by summary measures, but instead as partially independent symptoms that vary in prevalence across the lifespan. Better self-reported sleep is associated with better health outcomes, and the strength of these associations differs across health domains. Notably, we do observed associations between self-reported sleep quality and white matter.nnFundingBiotechnology and Biological Sciences Research Council (grant number BB/H008217/1). RAK is supported by the Wellcome Trust (grant number 107392/Z/15/Z and the UK Medical Research Council (MC-A060-5PR61).nnStrengths and limitations of this studyO_LIBroad phenotypic assessment of healthy ageing across multiple health domainsnC_LIO_LIAdvanced analytic techniques (i.e. Latent Class Analysis regression) allows new insightsnC_LIO_LIA uniquely large neuroimaging sample combined with Bayesian inference allows for quantification of evidence for the null hypothesisnC_LIO_LISubjective sleep measures may have drawbacks in older samplesnC_LIO_LICross-sectional data precludes modelling of within subject changesnC_LI
]]></description>
<dc:creator>Andrew Gadie</dc:creator>
<dc:creator>Meredith Shafto</dc:creator>
<dc:creator>Yue Leng</dc:creator>
<dc:creator>Cam-CAN</dc:creator>
<dc:creator>Rogier A. Kievit</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-06-22</dc:date>
<dc:identifier>doi:10.1101/060145</dc:identifier>
<dc:title><![CDATA[Age-related differences in self-reported sleep quality predict healthy ageing across multiple domains: a multi-modal cohort of 2406 adults]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/067678v1?rss=1">
<title>
<![CDATA[
Interactive online brain shape visualization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/067678v1?rss=1</link>
<description><![CDATA[
Urbanization presents unique environmental challenges to human commensal species. The Afrotropical Anopheles gambiae complex contains a number of synanthropic mosquito species that are major vectors of malaria. To examine ongoing cryptic diversification within the complex, we performed reduced representation sequencing on 941 mosquitoes collected across four ecogeographic zones in Cameroon. We find evidence for clear subdivision within An. coluzzii and An. gambiae s.s. - the two most significant malaria vectors in the region. Importantly, in both species rural and urban populations of mosquitoes were genetically differentiated. Genome scans of cryptic subgroups reveal pervasive signatures of selection centered on genes involved in xenobiotic resistance. Notably, a selective sweep containing eight detoxification enzymes is unique to urban mosquitoes that exploit polluted breeding sites. Overall, our study reveals that anthropogenic environmental modification is driving population differentiation and local adaptation in African malaria mosquitoes with potentially significant consequences for malaria epidemiology.
]]></description>
<dc:creator>Anisha Keshavan</dc:creator>
<dc:creator>Arno Klein</dc:creator>
<dc:creator>Benjamin Cipollini</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-08-05</dc:date>
<dc:identifier>doi:10.1101/067678</dc:identifier>
<dc:title><![CDATA[Interactive online brain shape visualization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/071860v1?rss=1">
<title>
<![CDATA[
Lost dollars threaten research in public academic health centers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/071860v1?rss=1</link>
<description><![CDATA[
Decreasing federal and state support threaten long-term sustainability of research in publicly supported academic health centers. In weathering these financial threats, research at the University of California, San Francisco (UCSF) has undergone three substantial changes: (i) institutional salary support goes preferentially to senior faculty, while the young increasingly depend on grants; (ii) private and government support for research grows apace in clinical departments, but slowly declines in basic science departments; and (iii) research is judged more on its quantity (numbers of investigators and federal and private dollars) than on its goals, achievements, or scientific quality. We propose measures to alleviate these problems. Other large public academic health centers probably confront similar issues, but--except for UCSF--such centers have not been subjected to detailed public analysis.
]]></description>
<dc:creator>Henry R. Bourne</dc:creator>
<dc:creator>Eric Vermillion</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-08-27</dc:date>
<dc:identifier>doi:10.1101/071860</dc:identifier>
<dc:title><![CDATA[Lost dollars threaten research in public academic health centers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/074864v1?rss=1">
<title>
<![CDATA[
Personalized genetic assessment of age associated Alzheimers disease risk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/074864v1?rss=1</link>
<description><![CDATA[
ImportanceIdentifying individuals at risk for developing Alzheimers disease (AD) is of utmost importance. Although genetic studies have identified APOE and other AD associated single nucleotide polymorphisms (SNPs), genetic information has not been integrated into an epidemiological framework for personalized risk prediction.nnObjectiveTo develop, replicate and validate a novel polygenic hazard score for predicting age-specific risk for AD.nnSettingMulti-center, multi-cohort genetic and clinical data.nnParticipantsWe assessed genetic data from 17,008 AD patients and 37,154 controls from the International Genetics of Alzheimers Project (IGAP), and 6,409 AD patients and 9,386 older controls from Phase 1 Alzheimers Disease Genetics Consortium (ADGC). As independent replication and validation cohorts, we also evaluated genetic, neuroimaging, neuropathologic, CSF and clinical data from ADGC Phase 2, National Institute of Aging Alzheimers Disease Center (NIA ADC) and Alzheimers Disease Neuroimaging Initiative (ADNI) (total n = 20,680)nnMain Outcome(s) and Measure(s)Use the IGAP cohort to first identify AD associated SNPs (at p < 10-5). Next, integrate these AD associated SNPs into a Cox proportional hazards model using ADGC phase 1 genetic data, providing a polygenic hazard score (PHS) for each participant. Combine population based incidence rates, and genotype-derived PHS for each individual to derive estimates of instantaneous risk for developing AD, based on genotype and age. Finally, assess replication and validation of PHS in independent cohorts.nnResultsIndividuals in the highest PHS quantiles developed AD at a considerably lower age and had the highest yearly AD incidence rate. Among APOE {varepsilon}3/3 individuals, PHS modified expected age of AD onset by more than 10 years between the lowest and highest deciles. In independent cohorts, PHS strongly predicted empirical age of AD onset (p = 1.1 x 10-26), longitudinal progression from normal aging to AD (p = 1.54 x 10-10) and associated with markers of AD neurodegeneration.nnConclusionsWe developed, replicated and validated a clinically usable PHS for quantifying individual differences in age-specific risk of AD. Beyond APOE, polygenic architecture plays an important role in modifying AD risk. Precise quantification of AD genetic risk will be useful for early diagnosis and therapeutic strategies.
]]></description>
<dc:creator>Rahul S. Desikan</dc:creator>
<dc:creator>Chun Chieh Fan</dc:creator>
<dc:creator>Yunpeng Wang</dc:creator>
<dc:creator>Andrew J. Schork</dc:creator>
<dc:creator>Howard J. Cabral</dc:creator>
<dc:creator>Adrienne Cupples</dc:creator>
<dc:creator>Wesley K. Thompson</dc:creator>
<dc:creator>Lilah Besser</dc:creator>
<dc:creator>Walter A. Kukull</dc:creator>
<dc:creator>Dominic Holland</dc:creator>
<dc:creator>Chi-Hua Chen</dc:creator>
<dc:creator>James B. Brewer</dc:creator>
<dc:creator>David S. Karow</dc:creator>
<dc:creator>Karolina Kauppi</dc:creator>
<dc:creator>Aree Witoelar</dc:creator>
<dc:creator>Celeste M. Karch</dc:creator>
<dc:creator>Luke W. Bonham</dc:creator>
<dc:creator>Jennifer S. Yokoyama</dc:creator>
<dc:creator>Howard J. Rosen</dc:creator>
<dc:creator>Bruce L. Miller</dc:creator>
<dc:creator>William P. Dillon</dc:creator>
<dc:creator>David M. Wilson</dc:creator>
<dc:creator>Christopher P. Hess</dc:creator>
<dc:creator>Margaret Pericak-Vance</dc:creator>
<dc:creator>Jonathan L. Haines</dc:creator>
<dc:creator>Lindsay A. Farrer</dc:creator>
<dc:creator>Richard Mayeux</dc:creator>
<dc:creator>John Hardy</dc:creator>
<dc:creator>Alison M. Goate</dc:creator>
<dc:creator>Bradley T. Hyman</dc:creator>
<dc:creator>Gerard D. Schellenberg</dc:creator>
<dc:creator>Linda K. McEvoy</dc:creator>
<dc:creator>Ole A. Andreassen</dc:creator>
<dc:creator>Anders N. Dale</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-09-13</dc:date>
<dc:identifier>doi:10.1101/074864</dc:identifier>
<dc:title><![CDATA[Personalized genetic assessment of age associated Alzheimers disease risk]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/076265v1?rss=1">
<title>
<![CDATA[
Adipocyte JAK2 mediates hepatic insulin sensitivity and the diabetogenic action of Growth Hormone 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/076265v1?rss=1</link>
<description><![CDATA[
For nearly 100 years, Growth Hormone (GH) has been known to impact insulin sensitivity and risk of diabetes. However, the tissue governing the effects of GH signaling on insulin and glucose homeostasis remains unknown. Excess GH reduces fat mass and insulin sensitivity. Conversely, GH insensitivity (GHI) is associated with increased adiposity, augmented insulin sensitivity, and protection from diabetes. Here we induce adipocyte-specific GHI through conditional deletion of Jak2 (JAK2A), an obligate transducer of GH signaling. Similar to whole-body GHI, JAK2A mice had increased adiposity and extreme insulin sensitivity. Loss of adipocyte Jak2 augmented hepatic insulin sensitivity and conferred resistance to diet-induced metabolic stress without overt changes in circulating fatty acids. While GH injections induced hepatic insulin resistance in control mice, the diabetogenic action was absent in JAK2A mice. Adipocyte GH signaling directly impinged on both adipose and hepatic insulin signal transduction. Collectively, our results show that adipose tissue governs the effects of GH on insulin and glucose homeostasis. Further, we show that JAK2 mediates liver insulin sensitivity via an extra-hepatic, adipose tissue-dependent mechanism.
]]></description>
<dc:creator>Kevin Corbit</dc:creator>
<dc:creator>Joao Paulo G. Camporez</dc:creator>
<dc:creator>Jennifer L Tran</dc:creator>
<dc:creator>Camella G Wilson</dc:creator>
<dc:creator>Rachel J Perry</dc:creator>
<dc:creator>Gerald Schulman</dc:creator>
<dc:creator>Michael J Jurczak</dc:creator>
<dc:creator>Ethan Weiss</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-09-20</dc:date>
<dc:identifier>doi:10.1101/076265</dc:identifier>
<dc:title><![CDATA[Adipocyte JAK2 mediates hepatic insulin sensitivity and the diabetogenic action of Growth Hormone]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/077917v1?rss=1">
<title>
<![CDATA[
AN INTERNATIONAL INTER-LABORATORY DIGITAL PCR STUDY DEMONSTRATES HIGH REPRODUCIBILITY FOR THE MEASUREMENT OF A RARE SEQUENCE VARIANT 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/077917v1?rss=1</link>
<description><![CDATA[
This study tested the claim that digital PCR (dPCR) can offer highly reproducible quantitative measurements in disparate labs. Twenty-one laboratories measured four blinded samples containing different quantities of a KRAS fragment encoding G12D, an important genetic marker for guiding therapy of certain cancers. This marker is challenging to quantify reproducibly using qPCR or NGS due to the presence of competing wild type sequences and the need for calibration. Using dPCR, eighteen laboratories were able to quantify the G12D marker within 12% of each other in all samples. Three laboratories appeared to measure consistently outlying results; however, proper application of a follow-up analysis recommendation rectified their data. Our findings show that dPCR has demonstrable reproducibility across a large number of laboratories without calibration and could enable the reproducible application of molecular stratification to guide therapy, and potentially for molecular diagnostics.nnSIGNIFICANCE STATEMENTThe poor reproducibility of molecular diagnostic methods limits their application in part due to the challenges associated with calibration of what are relative measurement approaches. In this study we investigate the performance of one of the only absolute measurement methods available today, digital PCR (dPCR), and demonstrated that when compared across twenty-one laboratories, dPCR has unprecedented reproducibility. These results were achieved when measuring a challenging single nucleotide variant and without calibration to any reference samples. This opens the possibility for dPCR to offer a method to transform reproducibility in the molecular diagnostic field, both by direct use as well as in support of other currently used clinical methods.
]]></description>
<dc:creator>Alexandra S Whale</dc:creator>
<dc:creator>Alison S Devonshire</dc:creator>
<dc:creator>George Karlin-Neumann</dc:creator>
<dc:creator>Jack Regan</dc:creator>
<dc:creator>Leanne Javier</dc:creator>
<dc:creator>Simon Cowen</dc:creator>
<dc:creator>Ana Fernandez-Gonzalez</dc:creator>
<dc:creator>Gerwyn M Jones</dc:creator>
<dc:creator>Nicholas Redshaw</dc:creator>
<dc:creator>Julia Beck</dc:creator>
<dc:creator>Andreas W Berger</dc:creator>
<dc:creator>Valerie Combaret</dc:creator>
<dc:creator>Nina Dahl Kjersgaard</dc:creator>
<dc:creator>Lisa Davis</dc:creator>
<dc:creator>Frederic Fina</dc:creator>
<dc:creator>Tim Forshew</dc:creator>
<dc:creator>Rikke Fredslund Andersen</dc:creator>
<dc:creator>Silvia Galbiati</dc:creator>
<dc:creator>Alvaro Gonzalez Hernandez</dc:creator>
<dc:creator>Charles Haynes</dc:creator>
<dc:creator>Filip Janku</dc:creator>
<dc:creator>Roger Lacave</dc:creator>
<dc:creator>Justin Lee</dc:creator>
<dc:creator>Vilas Mistry</dc:creator>
<dc:creator>Alexandra Pender</dc:creator>
<dc:creator>Anne Pradines</dc:creator>
<dc:creator>Charlotte Proudhon</dc:creator>
<dc:creator>Lao Saal</dc:creator>
<dc:creator>Elliot Stieglitz</dc:creator>
<dc:creator>Bryan Ulrich</dc:creator>
<dc:creator>Carole A Foy</dc:creator>
<dc:creator>Helen Parkes</dc:creator>
<dc:creator>Svilen Tzonev</dc:creator>
<dc:creator>Jim Huggett</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-09-28</dc:date>
<dc:identifier>doi:10.1101/077917</dc:identifier>
<dc:title><![CDATA[AN INTERNATIONAL INTER-LABORATORY DIGITAL PCR STUDY DEMONSTRATES HIGH REPRODUCIBILITY FOR THE MEASUREMENT OF A RARE SEQUENCE VARIANT]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/079145v1?rss=1">
<title>
<![CDATA[
BIDS Apps: Improving ease of use, accessibility and reproducibility of neuroimaging data analysis methods 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/079145v1?rss=1</link>
<description><![CDATA[
The rate of progress in human neurosciences is limited by the inability to easily apply a wide range of analysis methods to the plethora of different datasets acquired in labs around the world. In this work, we introduce a framework for creating, testing, versioning and archiving portable applications for analyzing neuroimaging data organized and described in compliance with the Brain Imaging Data Structure (BIDS). The portability of these applications (BIDS Apps) is achieved by using container technologies that encapsulate all binary and other dependencies in one convenient package. BIDS Apps run on all three major operating systems with no need for complex setup and configuration and thanks to the comprehensiveness richness of the BIDS standard they require little manual user input. Previous containerized data processing solutions were limited to single user environments and not compatible with most multi-tenant High Performance Computing systems. BIDS Apps overcome this limitation by taking advantage of the Singularity container technology. As a proof of concept, this work is accompanied by 22 ready to use BIDS Apps, packaging a diverse set of commonly used neuroimaging algorithms.nnAuthor SummaryMagnetic Resonance Imaging (MRI) is a non-invasive way to measure human brain structure and activity that has been used for over 25 years. There are thousands MRI studies performed every year generating a substantial amount of data. At the same time, many new data analysis methods are being developed every year. The potential of using new analysis methods on the variety of existing and newly acquired data is hindered by difficulties in software deployment and lack of support for standardized input data. Here we propose to use container technology to make deployment of a wide range of data analysis techniques easy. In addition, we adapt the existing data analysis tools to interface with data organized in a standardized way. We hope that this approach will enable researchers to access a wider range of methods when analyzing their data which will lead to accelerated progress in human neuroscience.
]]></description>
<dc:creator>Krzysztof J. Gorgolewski</dc:creator>
<dc:creator>Fidel Alfaro-Almagro</dc:creator>
<dc:creator>Tibor Auer</dc:creator>
<dc:creator>Pierre Bellec</dc:creator>
<dc:creator>Mihai Capota</dc:creator>
<dc:creator>Mallar Chakravarty</dc:creator>
<dc:creator>Nathan W. Churchill</dc:creator>
<dc:creator>R. Cameron Craddock</dc:creator>
<dc:creator>Gabriel Devenyi</dc:creator>
<dc:creator>Anders Eklund</dc:creator>
<dc:creator>Oscar Esteban</dc:creator>
<dc:creator>Guillaume Flandin</dc:creator>
<dc:creator>Satrajit Ghosh</dc:creator>
<dc:creator>J. Swaroop Guntupalli</dc:creator>
<dc:creator>Mark Jenkinson</dc:creator>
<dc:creator>Anisha Keshavan</dc:creator>
<dc:creator>Gregory Kiar</dc:creator>
<dc:creator>Pradeep Reddy Raamana</dc:creator>
<dc:creator>David Raffelt</dc:creator>
<dc:creator>Christopher J. Steele</dc:creator>
<dc:creator>Pierre-Olivier Quirion</dc:creator>
<dc:creator>Robert E. Smith</dc:creator>
<dc:creator>Stephen Strother</dc:creator>
<dc:creator>Gael Varoquaux</dc:creator>
<dc:creator>Tal Yarkoni</dc:creator>
<dc:creator>Yida Wang</dc:creator>
<dc:creator>Russell Poldrack</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-10-04</dc:date>
<dc:identifier>doi:10.1101/079145</dc:identifier>
<dc:title><![CDATA[BIDS Apps: Improving ease of use, accessibility and reproducibility of neuroimaging data analysis methods]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/079236v1?rss=1">
<title>
<![CDATA[
Disruption of Hepatocyte Jak2 leads to Spontaneous NASH in Aged Mice and Uncouples Metabolic Liver Disease from Insulin Resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/079236v1?rss=1</link>
<description><![CDATA[
Growth Hormone (GH) is a master regulator of metabolic homeostasis and longevity. Whole body GH insensitivity (GHI) augments insulin sensitivity, age-related disease resistance, adiposity, and occurrence of NAFLD. Conversely, acromegalic patients are prone to diabetes and increased mortality due to constitutive high levels of circulating GH. However, which tissues control the various metabolic aspects of GH physiology are unknown. Therefore, we determined the role of GH in age-related metabolic dysfunction by inducing hepatocyte- (JAK2L) or adipocyte-specific (JAK2A) GHI individually or combinatorially (JAK2LA) via deletion of Jak2, an obligate transducer of GH signaling. Aged JAK2L mice were insulin resistant but lean and had significant NASH, hepatic inflammation, and fibrosis. In contrast, JAK2A animals had increased adiposity and were completely resistant to age-associated hepatic steatosis, NASH, and insulin resistance. Interestingly, while JAK2LA mice retained enhanced whole-body insulin sensitivity, they still developed NASH to an almost identical degree as JAK2L mice but with a substantial reduction in the degree of microvesicular steatosis. Collectively, loss of adipocyte Jak2 conferred whole body insulin sensitivity even in the face of obesity and NASH. Deletion of hepatocyte Jak2 promoted NASH in aged mice without any dietary or drugs perturbations. The effect appears to be liver autonomous and cannot be overcome by the insulin sensitizing effect of adipocyte Jak2 deletion. Here, we describe the first model of spontaneous NASH that is coupled to augmented insulin sensitivity. Further, there was an inverse correlation between insulin sensitivity and the degree of microvesicular steatosis. Therefore, GH signaling independently mediates insulin/glucose and lipid homeostasis and directly regulates the development of NASH in aged mice.nnFinancial SupportThis study was supported by National Institutes of Health (NIH) Grants 1R01DK091276 (to E.J.W.). We also acknowledge the support of the University of California, San Francisco (UCSF) Cardiovascular Research Institute, the UCSF Diabetes Center (P30 DK063720), the UCSF Liver Center (P30 DK026743, and the James Peter Read Foundation.nnAbbreviations
]]></description>
<dc:creator>Camella G. Wilson</dc:creator>
<dc:creator>Jennifer L. Tran</dc:creator>
<dc:creator>Aras N. Mattis</dc:creator>
<dc:creator>Kevin Corbit</dc:creator>
<dc:creator>Ethan J. Weiss</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-10-04</dc:date>
<dc:identifier>doi:10.1101/079236</dc:identifier>
<dc:title><![CDATA[Disruption of Hepatocyte Jak2 leads to Spontaneous NASH in Aged Mice and Uncouples Metabolic Liver Disease from Insulin Resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/081646v1?rss=1">
<title>
<![CDATA[
Flexibility and design: conformational heterogeneity along the evolutionary trajectory of a redesigned ubiquitin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/081646v1?rss=1</link>
<description><![CDATA[
SummaryAlthough protein design has been used to introduce new functions, designed variants generally only function as well as natural proteins after rounds of laboratory evolution. One possibility for this pattern is that designed mutants frequently sample nonfunctional conformations. To test this idea, we exploited advances in multiconformer modeling of room temperature X-ray data collection on redesigned ubiquitin variants selected for increasing binding affinity to the deubiquitinase USP7. Initial core mutations disrupt natural packing and lead to increased flexibility. Additional, experimentally selected mutations quenched conformational heterogeneity through new stabilizing interactions. Stabilizing interactions, such as cation-pi stacking and ordered waters, which are not included in standard protein design energy functions, can create specific interactions that have long range effects on flexibility across the protein. Our results suggest that increasing flexibility may be a useful strategy to escape local minima during initial directed evolution and protein design steps when creating new functions.
]]></description>
<dc:creator>Biel, J. T.</dc:creator>
<dc:creator>Thompson, M. C.</dc:creator>
<dc:creator>Cunningham, C. N.</dc:creator>
<dc:creator>Corn, J. E.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2016-10-17</dc:date>
<dc:identifier>doi:10.1101/081646</dc:identifier>
<dc:title><![CDATA[Flexibility and design: conformational heterogeneity along the evolutionary trajectory of a redesigned ubiquitin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/084657v1?rss=1">
<title>
<![CDATA[
16S rRNA Gene Sequencing as a Clinical Diagnostic Aid for Gastrointestinal-related Conditions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/084657v1?rss=1</link>
<description><![CDATA[
Accurate detection of the microorganisms underlying gut dysbiosis in the patient is critical to initiate the appropriate treatment. However, most clinical microbiology techniques used to detect gut bacteria were developed over a century ago and rely on culture-based approaches that are often laborious, unreliable, and subjective. Further, culturing does not scale well for multiple targets and detects only a minority of the microorganisms in the human gastrointestinal tract. Here we present a clinical test for gut microorganisms based on targeted sequencing of the prokaryotic 16S rRNA gene. We tested 46 clinical prokaryotic targets in the human gut, 28 of which can be identified by a bioinformatics pipeline that includes sequence analysis and taxonomic annotation. Using microbiome samples from a cohort of 897 healthy individuals, we established a reference range defining clinically relevant relative levels for each of the 28 targets. Our assay accurately quantified all 28 targets and correctly reflected 38/38 verification samples of real and synthetic stool material containing known pathogens. Thus, we have established a new test to interrogate microbiome composition and diversity, which will improve patient diagnosis, treatment and monitoring. More broadly, our test will facilitate epidemiological studies of the microbiome as it relates to overall human health and disease.
]]></description>
<dc:creator>Almonacid, D. E.</dc:creator>
<dc:creator>Kraal, L.</dc:creator>
<dc:creator>Ossandon, F. J.</dc:creator>
<dc:creator>Budovskaya, Y. V.</dc:creator>
<dc:creator>Cardenas, J. P.</dc:creator>
<dc:creator>Richman, J.</dc:creator>
<dc:creator>Apte, Z. S.</dc:creator>
<dc:date>2016-10-31</dc:date>
<dc:identifier>doi:10.1101/084657</dc:identifier>
<dc:title><![CDATA[16S rRNA Gene Sequencing as a Clinical Diagnostic Aid for Gastrointestinal-related Conditions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/085233v1?rss=1">
<title>
<![CDATA[
Engineering Genetically-Encoded Mineralization and Magnetism via Directed Evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/085233v1?rss=1</link>
<description><![CDATA[
Genetically encoding the synthesis of functional nanomaterials such as magnetic nanoparticles enables sensitive and non-invasive biological sensing and control. Via directed evolution of the natural iron-sequestering ferritin protein, we discovered key mutations that lead to significantly enhanced cellular magnetism, resulting in increased physical attraction of ferritin-expressing cells to magnets and increased contrast for cellular magnetic resonance imaging (MRI). The magnetic mutants further demonstrate increased iron biomineralization measured by a novel fluorescent genetic sensor for intracellular free iron. In addition, we engineered Escherichia coli cells with multiple genomic knockouts to increase cellular accumulation of various metals. Lastly to explore further protein candidates for biomagnetism, we characterized members of the DUF892 family using the iron sensor and magnetic columns, confirming their intracellular iron sequestration that results in increased cellular magnetization.
]]></description>
<dc:creator>Liu, X. L.</dc:creator>
<dc:creator>Lopez, P. A.</dc:creator>
<dc:creator>Giessen, T. W.</dc:creator>
<dc:creator>Giles, M.</dc:creator>
<dc:creator>Way, J. C.</dc:creator>
<dc:creator>Silver, P. A.</dc:creator>
<dc:date>2016-11-02</dc:date>
<dc:identifier>doi:10.1101/085233</dc:identifier>
<dc:title><![CDATA[Engineering Genetically-Encoded Mineralization and Magnetism via Directed Evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/086553v1?rss=1">
<title>
<![CDATA[
Novel computational method for predicting polytherapy switching strategies to overcome tumor heterogeneity and evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/086553v1?rss=1</link>
<description><![CDATA[
1The success of targeted cancer therapy is limited by drug resistance that can result from tumor genetic heterogeneity. The current approach to address resistance typically involves initiating a new treatment after clinical/radiographic disease progression, ultimately resulting in futility in most patients. Towards a potential alternative solution, we developed a novel computational framework that uses human cancer profiling data to systematically identify dynamic, pre-emptive, and sometimes non-intuitive treatment strategies that can better control tumors in real-time. By studying lung adenocarcinoma clinical specimens and preclinical models, our computational analyses revealed that the best anti-cancer strategies addressed existing resistant subpopulations as they emerged dynamically during treatment. In some cases, the best computed treatment strategy used unconventional therapy switching while the bulk tumor was responding, a prediction we confirmed in vitro. The new framework presented here could guide the principled implementation of dynamic molecular monitoring and treatment strategies to improve cancer control.
]]></description>
<dc:creator>Jonsson, V. D.</dc:creator>
<dc:creator>Blakely, C. M.</dc:creator>
<dc:creator>Lin, L.</dc:creator>
<dc:creator>Asthana, S.</dc:creator>
<dc:creator>Olivas, V.</dc:creator>
<dc:creator>Gubens, M. A.</dc:creator>
<dc:creator>Matni, N.</dc:creator>
<dc:creator>Bastian, B. C.</dc:creator>
<dc:creator>Taylor, B. S.</dc:creator>
<dc:creator>Doyle, J. C.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:date>2016-11-08</dc:date>
<dc:identifier>doi:10.1101/086553</dc:identifier>
<dc:title><![CDATA[Novel computational method for predicting polytherapy switching strategies to overcome tumor heterogeneity and evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/086702v1?rss=1">
<title>
<![CDATA[
DNA-polymerase guided elimination of paternal mitochondrial genomes:An escape-proof obstacle to their transmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/086702v1?rss=1</link>
<description><![CDATA[
Mitochondrial DNA is predominantly inherited from only one parent. In animals this is usually the mother. This program is not in the interest of the paternal mitochondrial genome whose potential to contribute to future generations is restricted. However, in a dramatic example of genetic conflict, nuclear programs ensure the outcome. Two large mitochondria extend the length of Drosophila sperm tails. The hundreds of nucleoids in these mitochondria vanish during spermatogenesis eliminating their potential for transmission. Our previous work showed that mutational inactivation of EndoG, a nuclear encoded mitochondrial endonuclease, slows elimination of mitochondrial genomes. Here, we show that knockdown of the nuclearly encoded mitochondrial DNA polymerase, Tamas, produces a much more complete block of mtDNA loss. Recruitment of Tamas to the nucleoid at the time of its disappearance suggests a direct contribution to the elimination, but the 3'-exonuclease function of the polymerase is not needed. While DNA elimination is a surprising function for DNA polymerase, its use to restrict paternal genomes provides a strategy that cannot easily be evaded by the mitochondrial genome without compromising its replication.
]]></description>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>O'Farrell, P. H.</dc:creator>
<dc:creator>Yakubovich, N.</dc:creator>
<dc:creator>DeLuca, S. Z.</dc:creator>
<dc:date>2016-11-09</dc:date>
<dc:identifier>doi:10.1101/086702</dc:identifier>
<dc:title><![CDATA[DNA-polymerase guided elimination of paternal mitochondrial genomes:An escape-proof obstacle to their transmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/089235v1?rss=1">
<title>
<![CDATA[
Profiling adaptive immune repertoires across multiple human tissues by RNA Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/089235v1?rss=1</link>
<description><![CDATA[
Assay-based approaches provide a detailed view of the adaptive immune system by profiling T and B cell receptor repertoires. However, these methods come at a high cost and lack the scale of standard RNA sequencing (RNA-seq). Here we report the development of ImReP, a novel computational method for rapid and accurate profiling of the adaptive immune repertoire from regular RNA-Seq data. We applied it to 8,555 samples across 544 individuals from 53 tissues from the Genotype-Tissue Expression (GTEx v6) project. ImReP is able to efficiently extract TCR- and BCR- derived reads from the RNA-Seq data and accurately assemble the complementarity determining regions 3 (CDR3s), the most variable regions of B- and T-cell receptors determining their antigen specificity. Using ImReP, we have created the systematic atlas of immunological sequences for B- and T-cell repertoires across a broad range of tissue types, most of which have not been studied for B and T cell receptor repertoires. We have also examined the compositional similarities of clonal populations between the GTEx tissues to track the flow of T- and B- clonotypes across immune-related tissues, including secondary lymphoid organs and organs encompassing mucosal, exocrine, and endocrine sites. The atlas of T- and B-cell receptor receptors, freely available at https://sergheimangul.wordpress.com/atlas-immune-repertoires/, is the largest collection of CDR3 sequences and tissue types. We anticipate this recourse will enhance future studies in areas such as immunology and advance development of therapies for human diseases. ImReP is freely available at https://sergheimangul.wordpress.com/imrep/.
]]></description>
<dc:creator>Mangul, S.</dc:creator>
<dc:creator>Mandric, I.</dc:creator>
<dc:creator>Yang, H. T.</dc:creator>
<dc:creator>Strauli, N.</dc:creator>
<dc:creator>Montoya, D.</dc:creator>
<dc:creator>Rotman, J.</dc:creator>
<dc:creator>Der Wey, W. V.</dc:creator>
<dc:creator>Ronas, J. R.</dc:creator>
<dc:creator>Statz, B.</dc:creator>
<dc:creator>Zelikovsky, A.</dc:creator>
<dc:creator>Spreafico, R.</dc:creator>
<dc:creator>Shifman, S.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:creator>Rossetti, M.</dc:creator>
<dc:creator>Ansel, K. M.</dc:creator>
<dc:creator>Eskin, E.</dc:creator>
<dc:date>2016-11-22</dc:date>
<dc:identifier>doi:10.1101/089235</dc:identifier>
<dc:title><![CDATA[Profiling adaptive immune repertoires across multiple human tissues by RNA Sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/090241v1?rss=1">
<title>
<![CDATA[
Genetic determinants of chromatin accessibility and gene regulation in T cell activation across human individuals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/090241v1?rss=1</link>
<description><![CDATA[
Over 90% of genetic variants associated with complex human traits map to non-coding regions, but little is understood about how they modulate gene regulation in health and disease. One possible mechanism is that genetic variants affect the activity of one or more cis-regulatory elements leading to gene expression variation in specific cell types. To identify such cases, we analyzed Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq) and RNA-seq profiles from activated CD4+ T cells of up to 105 healthy donors. We found that regions of accessible chromatin (ATAC-peaks) are co-accessible at kilobase and megabase resolution, in patterns consistent with the 3D organization of chromosomes measured by in situ Hi-C in T cells. 15% of genetic variants located within ATAC-peaks affected the accessibility of the corresponding peak through disrupting binding sites for transcription factors important for T cell differentiation and activation. These ATAC quantitative trait nucleotides (ATAC-QTNs) have the largest effects on co-accessible peaks, are associated with gene expression from the same aliquot of cells, are rarely affecting core binding motifs, and are enriched for autoimmune disease variants. Our results provide insights into how natural genetic variants modulate cis- regulatory elements, in isolation or in concert, to influence gene expression in primary immune cells that play a key role in many human diseases.
]]></description>
<dc:creator>Cheng, C. S.</dc:creator>
<dc:creator>Gate, R. E.</dc:creator>
<dc:creator>Aiden, A. P.</dc:creator>
<dc:creator>Siba, A.</dc:creator>
<dc:creator>Tabaka, M.</dc:creator>
<dc:creator>Lituiev, D.</dc:creator>
<dc:creator>Machol, I.</dc:creator>
<dc:creator>Subramaniam, M.</dc:creator>
<dc:creator>Shammim, M.</dc:creator>
<dc:creator>Hougen, K. L.</dc:creator>
<dc:creator>Wortman, I.</dc:creator>
<dc:creator>Huang, S.-C.</dc:creator>
<dc:creator>Durand, N. C.</dc:creator>
<dc:creator>Feng, T.</dc:creator>
<dc:creator>De Jager, P. L.</dc:creator>
<dc:creator>Chang, H. Y.</dc:creator>
<dc:creator>Lieberman Aiden, E.</dc:creator>
<dc:creator>Benoist, C.</dc:creator>
<dc:creator>Beer, M. A.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Regev, A.</dc:creator>
<dc:date>2016-12-02</dc:date>
<dc:identifier>doi:10.1101/090241</dc:identifier>
<dc:title><![CDATA[Genetic determinants of chromatin accessibility and gene regulation in T cell activation across human individuals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/090431v1?rss=1">
<title>
<![CDATA[
Mindcontrol: A Web Application for Brain Segmentation Quality Control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/090431v1?rss=1</link>
<description><![CDATA[
Tissue classification plays a crucial role in the investigation of normal neural development, brain-behavior relationships, and the disease mechanisms of many psychiatric and neurological illnesses. Ensuring the accuracy of tissue classification is important for quality research and, in particular, the translation of imaging biomarkers to clinical practice. Assessment with the human eye is vital to correct various errors inherent to all currently available segmentation algorithms. Manual quality assurance becomes methodologically difficult at a large scale - a problem of increasing importance as the number of data sets is on the rise. To make this process more efficient, we have developed Mindcontrol, an open-source web application for the collaborative quality control of neuroimaging processing outputs. The Mindcontrol platform consists of a dashboard to organize data, descriptive visualizations to explore the data, an imaging viewer, and an in-browser annotation and editing toolbox for data curation and quality control. Mindcontrol is flexible and can be configured for the outputs of any software package in any data organization structure. Example configurations for three large, open-source datasets are presented: the 1000 Functional Connectomes Project (FCP), the Consortium for Reliability and Reproducibility (CoRR), and the Autism Brain Imaging Data Exchange (ABIDE) Collection. These demo applications link descriptive quality control metrics, regional brain volumes, and thickness scalars to a 3D imaging viewer and editing module, resulting in an easy-to-implement quality control protocol that can be scaled for any size and complexity of study.
]]></description>
<dc:creator>Keshavan, A.</dc:creator>
<dc:creator>Datta, E.</dc:creator>
<dc:creator>McDonough, I.</dc:creator>
<dc:creator>Madan, C. R.</dc:creator>
<dc:creator>Jordan, K.</dc:creator>
<dc:creator>Henry, R.</dc:creator>
<dc:date>2016-11-29</dc:date>
<dc:identifier>doi:10.1101/090431</dc:identifier>
<dc:title><![CDATA[Mindcontrol: A Web Application for Brain Segmentation Quality Control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/091603v1?rss=1">
<title>
<![CDATA[
Genomic characterization of serial-passaged Ebola virus in a boa constrictor cell line 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/091603v1?rss=1</link>
<description><![CDATA[
Ebola virus disease (EVD) is a viral hemorrhagic fever with a high case-fatality rate in humans. EVD is caused by four members of the filoviral genus Ebolavirus, with Ebola virus (EBOV) being the most notorious one. Although bats are discussed as potential ebolavirus reservoirs, limited data actually support this hypothesis. Glycoprotein 2 (GP2) of reptarenaviruses, known to infect only boa constrictors and pythons, are similar in sequence and structure to ebolaviral glycoprotein 2 (GP2), suggesting that EBOV may be able to infect snake cells. We therefore serially passaged EBOV and a distantly related filovirus, Marburg virus (MARV), in the boa constrictor kidney cell line, JK, and characterized viral growth and mutational frequency by sequencing. We observed that EBOV efficiently infected and replicated in JK cells, but MARV did not. In contrast to most cell lines, EBOV infected JK cells did not result in obvious cytopathic effect (CPE). Genomic characterization of serial-passaged EBOV in JK cells revealed that genomic adaptation was not required for infection. Deep sequencing coverage (>10,000x) demonstrated the existence of only a single non-synonymous variant (EBOV glycoprotein precursor preGP T544I) of unknown significance within the viral population that exhibited a shift in frequency of at least 10% over six passages. Our data suggest that boid snake derived cells are competent for filovirus infection without appreciable genomic adaptation; that cellular filovirus infection without CPE may be more common than currently appreciated; and that there may be significant differences between the natural host spectra of ebolaviruses and marburgviruses.nnIMPORTANCEEbola virus (EBOV) causes a high case-fatality form of viral hemorrhagic fever. The natural reservoir of EBOV remains unknown. EBOV is distantly related to Marburg virus (MARV), which has been found in bats in the wild. The glycoprotein of a reptarenavirus known to infect boid snakes (pythons and boas) shows similarity in sequence and structure to these viruses, suggesting that EBOV and MARV may be able to infect and replicate in snake cells. We demonstrate that JK, a boa constrictor cell line, does not support MARV infection, but does support EBOV infection without causing overt cytopathic effect or the need for appreciable adaptation. These findings suggest different filoviruses may have a more diverse natural host spectra than previously thought.
]]></description>
<dc:creator>Fedewa, G.</dc:creator>
<dc:creator>Radoshitzky, S. R.</dc:creator>
<dc:creator>Chī, X.</dc:creator>
<dc:creator>Dŏng, L.</dc:creator>
<dc:creator>Spear, M.</dc:creator>
<dc:creator>Strauli, N.</dc:creator>
<dc:creator>Stenglein, M. D.</dc:creator>
<dc:creator>Hernandez, R.</dc:creator>
<dc:creator>Jahrling, P. B.</dc:creator>
<dc:creator>Kuhn, J. H.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:date>2016-12-07</dc:date>
<dc:identifier>doi:10.1101/091603</dc:identifier>
<dc:title><![CDATA[Genomic characterization of serial-passaged Ebola virus in a boa constrictor cell line]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/098731v1?rss=1">
<title>
<![CDATA[
MinorityReport, software for generalized analysis of causal genetic variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/098731v1?rss=1</link>
<description><![CDATA[
BackgroundThe widespread availability of next generation genome sequencing technologies has enabled a wide range of variant detection applications, especially in cancer and inborn genetic disorders. For model systems and microorganisms, the same technology may be used to discover the causative mutations for any phenotype, including those generated in response to chemical perturbation. In the case of pathogenic organisms, these approaches have allowed the determination of drug targets by means of resistance selection followed by genome sequencing.nnResultsHere, we present open source software written in python, MinorityReport, to facilitate the comparison of any two sets of genome alignments for the purpose of rapidly identifying the spectrum of nonsynonymous changes, insertions or deletions, and copy number variations in a presumed mutant relative to its parent. Specifically, MinorityReport relates mapped sequence reads in SAM format output from any alignment tool for both the mutant and parent genome, relative to a reference genome, and produces the set of variants that distinguishes the mutant from the parent, all presented in an intuitive, straightforward report format. MinorityReport features tunable parameters for evaluating evidence and a scoring system that prioritizes reported variants based on relative proportions of read counts supporting the variant in the mutant versus parent data sets. We demonstrate the utility of MinorityReport using publicly available data sets that we previously published to find the determinants of resistance for novel anti-malarial drugs.nnConclusionsMinorityReport is readily available (github: xxxxxxx) to identify the genetic mechanisms of drug resistance in plasmodium, genotype-phenotype relationships in human diads, or genomic variations between any two related organisms.
]]></description>
<dc:creator>Horst, J.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:date>2017-01-06</dc:date>
<dc:identifier>doi:10.1101/098731</dc:identifier>
<dc:title><![CDATA[MinorityReport, software for generalized analysis of causal genetic variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/099598v1?rss=1">
<title>
<![CDATA[
Local Control of Intracellular Microtubule Dynamics by End Binding Protein 1 (EB1) Photo-Dissociation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/099598v1?rss=1</link>
<description><![CDATA[
Dynamic remodelling of the microtubule cytoskeleton and local interactions with intracellular targets are central to many polarized cell biological processes, an idea first formalized as search-and-capture hypothesis three decades ago1. However, because of the rapid timescale of microtubule polymerization dynamics, it is difficult to directly ask how, when and where dynamic microtubules participate in specific biological processes. Here, we employ a blue light-sensitive interaction with the oat phototropin LOV2 domain2 to generate a photo-inactivated variant of the microtubule end-binding protein EB1, a small adaptor that is central to the interaction of functionally and structurally diverse proteins with growing microtubule ends3,4, that can replace endogenous EB1 function. Acute and reversible blue light-mediated n-EB1 photo-dissociation allows spatially and temporally precise control of intracellular microtubule polymerization dynamics. In addition to demonstrating that neither the GTP cap nor the MT polymerase CKAP5 are sufficient to sustain persistent MT polymerization at physiological growth rates, our data illustrate accurate subcellular control of a freely diffusible, cytoplasmic protein at the second and micrometer scale. This novel design may serve as a template for precise control of many other intracellular protein activities.
]]></description>
<dc:creator>van Haren, J.</dc:creator>
<dc:creator>Ettinger, A.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Hahn, K.</dc:creator>
<dc:creator>Wittmann, T.</dc:creator>
<dc:date>2017-01-10</dc:date>
<dc:identifier>doi:10.1101/099598</dc:identifier>
<dc:title><![CDATA[Local Control of Intracellular Microtubule Dynamics by End Binding Protein 1 (EB1) Photo-Dissociation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/100651v1?rss=1">
<title>
<![CDATA[
The design principles of biochemical timers: circuits that discriminate between transient and sustained stimulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/100651v1?rss=1</link>
<description><![CDATA[
Many cellular responses for which timing is critical display temporal filtering - the ability to suppress response until stimulated for longer than a given minimal time. Temporal filtering can play a key role in filtering noise, choreographing the timing of events, and mediating the interpretation of dynamically encoded signals. To define the biochemical circuits capable of kinetic filtering, we comprehensively searched the space of three-node networks. We define a metric of "temporal ultrasensitivity", a measure of the steepness of activation as a function of stimulus duration. We identified five classes of core network motifs capable of temporal filtering, each with different functional properties such as rejecting high frequency noise, committing to response (bistability), and distinguishing between long stimuli. Combinations of the two most robust motifs, double inhibition (DI) and positive feedback with AND logic (PFAND), underlie several natural timer circuits involved in processes such as cell cycle transitions, T cell activation, and departure from the pluripotent state. The biochemical network motifs described in this study form a basis for understanding the common ways in which cells make dynamic decisions.
]]></description>
<dc:creator>Gerardin, J.</dc:creator>
<dc:creator>Lim, W. A.</dc:creator>
<dc:date>2017-01-15</dc:date>
<dc:identifier>doi:10.1101/100651</dc:identifier>
<dc:title><![CDATA[The design principles of biochemical timers: circuits that discriminate between transient and sustained stimulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/101444v1?rss=1">
<title>
<![CDATA[
Tracking multiple genomic elements using correlative CRISPR imaging and sequential DNA FISH 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/101444v1?rss=1</link>
<description><![CDATA[
Live imaging of genome has offered important insights into the dynamics of the genome organization and gene expression. The demand to image simultaneously multiple genomic loci has prompted a flurry of exciting advances in multi-color CRISPR imaging, although color-based multiplexing is limited by the need for spectrally distinct fluorophores. Here we introduce an approach to achieve highly multiplexed live recording via correlative CRISPR imaging and sequential DNA fluorescence in situ hybridization (FISH). This approach first performs one-color live imaging of multiple genomic loci and then uses sequential rounds of DNA FISH to determine the loci identity. We have optimized the FISH protocol so that each round is complete in 1 min, demonstrating the identification of 7 genomic elements and the capability to sustain reversible staining and washing for up to 20 rounds. We have also developed a correlation-based algorithm to faithfully register live and FISH images. Our approach keeps the rest of the color palette open to image other cellular phenomena of interest, as demonstrated by our simultaneous live imaging of genomic loci together with a cell cycle reporter. Furthermore, the algorithm to register faithfully between live and fixed imaging is directly transferrable to other systems such as multiplex RNA imaging with RNA-FISH and multiplex protein imaging with antibody-staining.
]]></description>
<dc:creator>Guan, J.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Shi, X.</dc:creator>
<dc:creator>Feng, S.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:date>2017-01-18</dc:date>
<dc:identifier>doi:10.1101/101444</dc:identifier>
<dc:title><![CDATA[Tracking multiple genomic elements using correlative CRISPR imaging and sequential DNA FISH]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/102673v1?rss=1">
<title>
<![CDATA[
Locomotion enhances neural encoding of visual stimuli in mouse V1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/102673v1?rss=1</link>
<description><![CDATA[
Neurons in mouse primary visual cortex (V1) are selective for particular properties of visual stimuli. Locomotion causes a change in cortical state that leaves their selectivity unchanged but strengthens their responses. Both locomotion and the change in cortical state are initiated by projections from the mesencephalic locomotor region (MLR), the latter through a disinhibitory circuit in V1. The function served by this change in cortical state is unknown. By recording simultaneously from a large number of single neurons in alert mice viewing moving gratings, we investigated the relationship between locomotion and the information contained within the neural population. We found that locomotion improved encoding of visual stimuli in V1 by two mechanisms. First, locomotion-induced increases in firing rates enhanced the mutual information between visual stimuli and single neuron responses over a fixed window of time. Second, stimulus discriminability was improved, even for fixed population firing rates, because of a decrease in noise correlations across the population during locomotion. These two mechanisms contributed differently to improvements in discriminability across cortical layers, with changes in firing rates most important in the upper layers and changes in noise correlations most important in layer V. Together, these changes resulted in a three- to five-fold reduction in the time needed to precisely encode grating direction and orientation. These results support the hypothesis that cortical state shifts during locomotion to accommodate an increased load on the visual system when mice are moving.nnSignificance StatementThis paper contains three novel findings about the representation of information in neurons within the primary visual cortex of the mouse. First, we show that locomotion reduces by at least a factor of three the time needed for information to accumulate in the visual cortex that allows the distinction of different visual stimuli. Second, we show that the effect of locomotion is to increase information in cells of all layers of the visual cortex. Third we show that the means by which information is enhanced by locomotion differs between the upper layers, where the major effect is the increasing of firing rates, and in layer V, where the major effect is the reduction in noise correlations.
]]></description>
<dc:creator>Dadarlat, M. C.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:date>2017-01-24</dc:date>
<dc:identifier>doi:10.1101/102673</dc:identifier>
<dc:title><![CDATA[Locomotion enhances neural encoding of visual stimuli in mouse V1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/103572v1?rss=1">
<title>
<![CDATA[
Mapping load-bearing in the mammalian spindle reveals local kinetochore-fiber anchorage that provides mechanical isolation and redundancy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/103572v1?rss=1</link>
<description><![CDATA[
Active forces generated at kinetochores move chromosomes, and the dynamic spindle must robustly anchor kinetochore-fibers (k-fibers) to bear this load. We know that the mammalian spindle body can bear the load of chromosome movement far from poles, but do not know where and how - physically and molecularly - this load is distributed across the spindle. In part, this is because perturbing and reading out spindle mechanics in live cells is difficult. Yet, answering this question is key to understanding how the spindle generates and responds to force, and performs its diverse mechanical functions. Here, we map load-bearing across the mammalian spindle in space-time, and dissect local anchorage mechanics and mechanism. To do so, we laser ablate single k-fibers at different spindle locations, and in different molecular backgrounds, and quantify at high time resolution the immediate relaxation of chromosomes, k-fibers, and microtubule speckles. We find that load redistribution is locally confined in all directions: along the first 3-4 m from kinetochores, scaling with k-fiber length, and laterally within ~2 m of k-fiber sides, without neighboring k-fibers sharing load. A phenomenological model constrains the mechanistic underpinnings of these data: it suggests that dense, transient crosslinks to the spindle along k-fibers bear the load of chromosome movement, but that these connections do not limit the timescale of spindle reorganization. The microtubule crosslinker NuMA is needed for the local load-bearing observed, while Eg5 and PRC1 are not, suggesting specialization in mechanical function and a novel function for NuMA throughout the spindle body. Together, the data and model suggest that widespread NuMA-mediated crosslinks locally bear load, providing mechanical isolation and redundancy while allowing spindle fluidity. These features are well-suited to support robust chromosome segregation.
]]></description>
<dc:creator>Elting, M. W.</dc:creator>
<dc:creator>Prakash, M.</dc:creator>
<dc:creator>Udy, D. B.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2017-01-26</dc:date>
<dc:identifier>doi:10.1101/103572</dc:identifier>
<dc:title><![CDATA[Mapping load-bearing in the mammalian spindle reveals local kinetochore-fiber anchorage that provides mechanical isolation and redundancy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/107094v1?rss=1">
<title>
<![CDATA[
Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/107094v1?rss=1</link>
<description><![CDATA[
Hsp90 is a homodimeric ATP-dependent molecular chaperone that remodels its substrate "client" proteins, facilitating their folding and activating them for biological function. Despite decades of research, the mechanism connecting ATP hydrolysis and chaperone function remains elusive. Particularly puzzling has been the apparent lack of cooperativity in hydrolysis of the ATP in each protomer. A crystal structure of the mitochondrial Hsp90, TRAP1, revealed that the catalytically active state is closed in a highly-strained asymmetric conformation. This asymmetry, unobserved in other Hsp90 homologs, is due to buckling of one of the protomers and is most pronounced at the broadly conserved client-binding region. Here, we show that rather than being cooperative or independent, ATP hydrolysis on the two protomers is sequential and deterministic. Moreover, dimer asymmetry sets up differential hydrolysis rates for each protomer, such that the buckled conformation favors ATP hydrolysis. Remarkably, after the first hydrolysis, the dimer undergoes a flip in the asymmetry while remaining in a closed state for the second hydrolysis. From these results, we propose a model where direct coupling of ATP hydrolysis and conformational flipping rearranges client-binding sites, providing a paradigm of how energy from ATP hydrolysis can be used for client remodeling.
]]></description>
<dc:creator>Elnatan, D.</dc:creator>
<dc:creator>Betegon, M.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Ramelot, T.</dc:creator>
<dc:creator>Kennedy, M. A.</dc:creator>
<dc:creator>Agard, D.</dc:creator>
<dc:date>2017-02-09</dc:date>
<dc:identifier>doi:10.1101/107094</dc:identifier>
<dc:title><![CDATA[Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/107441v1?rss=1">
<title>
<![CDATA[
Distinct neural systems for the production of communicative vocal behaviors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/107441v1?rss=1</link>
<description><![CDATA[
Vocalizations are the production of sounds by the coordinated activity of up to eighty respiratory and laryngeal muscles. Whilst voiced acts, modified by the upper vocal tract (tongue, jaw, lip and palate) are central to the production of human speech, they are also central to the production of emotional vocalizations such as sounds of disgust, anger, laughter and crying. Evidence suggests that the speech and emotional vocalizations may comprise distinct classes of vocal movements: patients with speech production deficits are often able to produce emotional vocalizations. In addition to this, the ontogeny of the two articulations is largely distinct, with some culturally universal emotional vocalizations emerging soon after birth and human speech being a culture specific, highly learnt motor act, which must develop to some degree before a critical period in development. Here we investigate the motor pathways underlying these two classes of vocal movements for the first time by directly comparing BOLD responses during production of speech and non-speech vocal movements. Using functional magnetic resonance imaging, we report distinct patterns of activity in both subcortical and cortical regions (putamen and bilateral inferior frontal and parietal cortices) during the production of emotional vocalizations compared to speech production. In contrast we show that responses in primary sensorimotor regions do not differ during the production of speech and emotional vocalizations, suggesting partially overlapping, and partially non-overlapping neural structures for the motor control of these two classes of movement. In addition to this we report that responses in auditory cortices are distinct during the production of speech and non-speech vocalizations, suggesting that feedback control of speech and emotional vocalizations are distinct. These data provide novel evidence for the presence of dual pathways for the neural control of complex articulatory movements in humans. These findings are discussed in relation to the clinical and primate literature of vocal motor control.nnSignificance StatementThis work marks the first evidence in healthy humans for dual routes to vocal behaviors. Clinical evidence suggests that patients unable to produce speech may still be able to produce other vocal behaviors that employ the same set of effectors. Here we demonstrate that the production of different classes of vocal behavior is associated with overlapping and distinct networks of activity. Moreover, we show that auditory processing that occurs during the production of these movements may be distinct, suggesting that feedback may be used differently for these distinct classes of vocal movement.
]]></description>
<dc:creator>Agnew, Z. K.</dc:creator>
<dc:creator>Ward, L.</dc:creator>
<dc:creator>McGettigan, C.</dc:creator>
<dc:creator>Josephs, O.</dc:creator>
<dc:creator>Scott, S. K.</dc:creator>
<dc:date>2017-02-17</dc:date>
<dc:identifier>doi:10.1101/107441</dc:identifier>
<dc:title><![CDATA[Distinct neural systems for the production of communicative vocal behaviors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/107573v1?rss=1">
<title>
<![CDATA[
Duplication events downstream of IRX1 cause North Carolina macular dystrophy at the MCDR3 locus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/107573v1?rss=1</link>
<description><![CDATA[
Autosomal dominant North Carolina macular dystrophy (NCMD) is believed to represent a failure of macular development. The disorder has been assigned by linkage to two loci, MCDR1 on chromosome 6q16 and MCDR3 on chromosome 5p15-p13. Recently, noncoding variants upstream of PRDM13 and a large duplication including IRX1 have been identified. However, the underlying disease-causing mechanism remains uncertain. Through a combination of sequencing studies, we report two novel overlapping duplications at the MCDR3 locus, in a gene desert downstream of IRX1 and upstream of ADAMTS16. One duplication of 43 kb was identified in nine NCMD families (with evidence for a shared ancestral haplotype), and another one of 45 kb was found in a single family. The MCDR3 locus is thus refined to a shared region of 39 kb that contains DNAse hypersensitive sites active at a restricted time window during retinal development. Publicly available data confirmed expression of IRX1 and ADAMTS16 in human fetal retina, with IRX1 preferentially expressed in fetal macula. These findings represent a major advance in our understanding of the molecular genetics of NCMD at the MCDR3 locus and provide insights into the genetic pathways involved in human macular development.nnAbbreviations list
]]></description>
<dc:creator>Cipriani, V.</dc:creator>
<dc:creator>Silva, R. S.</dc:creator>
<dc:creator>Arno, G.</dc:creator>
<dc:creator>Pontikos, N.</dc:creator>
<dc:creator>Kalhoro, A.</dc:creator>
<dc:creator>Valeina, S.</dc:creator>
<dc:creator>Inashkina, I.</dc:creator>
<dc:creator>Audere, M.</dc:creator>
<dc:creator>Rutka, K.</dc:creator>
<dc:creator>Puech, B.</dc:creator>
<dc:creator>Michaelides, M.</dc:creator>
<dc:creator>van Heyningen, V.</dc:creator>
<dc:creator>Lace, B.</dc:creator>
<dc:creator>Webster, A. R. R.</dc:creator>
<dc:creator>Moore, A. T.</dc:creator>
<dc:date>2017-02-12</dc:date>
<dc:identifier>doi:10.1101/107573</dc:identifier>
<dc:title><![CDATA[Duplication events downstream of IRX1 cause North Carolina macular dystrophy at the MCDR3 locus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/117291v1?rss=1">
<title>
<![CDATA[
Evolution and clinical impact of genetic epistasis within EGFR-mutant lung cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/117291v1?rss=1</link>
<description><![CDATA[
Introductory paragraph Introductory paragraph Main text Methods Author Contributions Author Information References The current understanding of tumorigenesis is largely centered on a monogenic driver oncogene model. This paradigm is incompatible with the prevailing clinical experience in most solid malignancies: monotherapy with a drug directed against an individual oncogenic driver typically results in incomplete clinical responses and eventual tumor progression1-7. By profiling the somatic genetic alterations present in over 2,000 cases of lung cancer, the leading cause of cancer mortality worldwide8,9, we show that combinations of functional genetic alterations, i.e. genetic collectives dominate the landscape of ...
]]></description>
<dc:creator>Blakely, C. M.</dc:creator>
<dc:creator>Watkins, T. B. K.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Gini, B.</dc:creator>
<dc:creator>Chabon, J. J.</dc:creator>
<dc:creator>McCoach, C. E.</dc:creator>
<dc:creator>McGranahan, N.</dc:creator>
<dc:creator>Wilson, G. A.</dc:creator>
<dc:creator>Birkbak, N. J.</dc:creator>
<dc:creator>Olivas, V.</dc:creator>
<dc:creator>Rotow, J.</dc:creator>
<dc:creator>Maynard, A.</dc:creator>
<dc:creator>Wang, V.</dc:creator>
<dc:creator>Gubens, M.</dc:creator>
<dc:creator>Banks, K.</dc:creator>
<dc:creator>Lanman, R.</dc:creator>
<dc:creator>Caulin, A.</dc:creator>
<dc:creator>St. John, J.</dc:creator>
<dc:creator>Cordero, A.</dc:creator>
<dc:creator>Giannikopoulos, P.</dc:creator>
<dc:creator>Mack, P.</dc:creator>
<dc:creator>Gandara, D.</dc:creator>
<dc:creator>Husain, H.</dc:creator>
<dc:creator>Doebele, R.</dc:creator>
<dc:creator>Riess, J.</dc:creator>
<dc:creator>Diehn, M.</dc:creator>
<dc:creator>Swanton, C.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:date>2017-03-16</dc:date>
<dc:identifier>doi:10.1101/117291</dc:identifier>
<dc:title><![CDATA[Evolution and clinical impact of genetic epistasis within EGFR-mutant lung cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/118778v1?rss=1">
<title>
<![CDATA[
Multiplexing droplet-based single cell RNA-sequencing using natural genetic barcodes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/118778v1?rss=1</link>
<description><![CDATA[
Droplet-based single-cell RNA-sequencing (dscRNA-seq) has enabled rapid, massively parallel profiling of transcriptomes from tens of thousands of cells. Multiplexing samples for single cell capture and library preparation in dscRNA-seq would enable cost-effective designs of differential expression and genetic studies while avoiding technical batch effects, but its implementation remains challenging. Here, we introduce an in-silico algorithm demuxlet that harnesses natural genetic variation to discover the sample identity of each cell and identify droplets containing two cells. These capabilities enable multiplexed dscRNA-seq experiments where cells from unrelated individuals are pooled and captured at higher throughput than standard workflows. To demonstrate the performance of demuxlet, we sequenced 3 pools of peripheral blood mononuclear cells (PBMCs) from 8 lupus patients. Given genotyping data for each individual, demuxlet correctly recovered the sample identity of > 99% of singlets, and identified doublets at rates consistent with previous estimates. In PBMCs, we demonstrate the utility of multiplexed dscRNA-seq in two applications: characterizing cell type specificity and inter-individual variability of cytokine response from 8 lupus patients and mapping genetic variants associated with cell type specific gene expression from 23 donors. Demuxlet is fast, accurate, scalable and could be extended to other single cell datasets that incorporate natural or synthetic DNA barcodes.
]]></description>
<dc:creator>Kang, H. M.</dc:creator>
<dc:creator>Subramaniam, M.</dc:creator>
<dc:creator>Targ, S.</dc:creator>
<dc:creator>Nguyen, M.</dc:creator>
<dc:creator>Maliskova, L.</dc:creator>
<dc:creator>Wan, E.</dc:creator>
<dc:creator>Wong, S.</dc:creator>
<dc:creator>Byrnes, L.</dc:creator>
<dc:creator>Lanata, C.</dc:creator>
<dc:creator>Gate, R.</dc:creator>
<dc:creator>Mostafavi, S.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:creator>Criswell, L. A.</dc:creator>
<dc:creator>Ye, J.</dc:creator>
<dc:date>2017-03-20</dc:date>
<dc:identifier>doi:10.1101/118778</dc:identifier>
<dc:title><![CDATA[Multiplexing droplet-based single cell RNA-sequencing using natural genetic barcodes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/121020v1?rss=1">
<title>
<![CDATA[
Towards Human Super EEG 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/121020v1?rss=1</link>
<description><![CDATA[
We present a model-based method for inferring full-brain neural activity at millimeter-scale spatial resolutions and millisecond-scale temporal resolutions using standard human intracranial recordings. Our approach makes the simplifying assumptions that different peoples brains exhibit similar correlational structure, and that activity and correlation patterns vary smoothly over space. One can then ask, for an arbitrary individuals brain: given recordings from a limited set of locations in that individuals brain, along with the observed spatial correlations learned from other peoples recordings, how much can be inferred about ongoing activity at other locations throughout that individuals brain? We show that our approach generalizes across people and tasks, thereby providing a person- and task-general means of inferring high spatiotemporal resolution full-brain neural dynamics from standard low-density intracranial recordings.
]]></description>
<dc:creator>Owen, L. L. W.</dc:creator>
<dc:creator>Manning, J. R.</dc:creator>
<dc:date>2017-03-27</dc:date>
<dc:identifier>doi:10.1101/121020</dc:identifier>
<dc:title><![CDATA[Towards Human Super EEG]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/121202v1?rss=1">
<title>
<![CDATA[
The Human Cell Atlas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/121202v1?rss=1</link>
<description><![CDATA[
The recent advent of methods for high-throughput single-cell molecular profiling has catalyzed a growing sense in the scientific community that the time is ripe to complete the 150-year-old effort to identify all cell types in the human body, by undertaking a Human Cell Atlas Project as an international collaborative effort. The aim would be to define all human cell types in terms of distinctive molecular profiles (e.g., gene expression) and connect this information with classical cellular descriptions (e.g., location and morphology). A comprehensive reference map of the molecular state of cells in healthy human tissues would propel the systematic study of physiological states, developmental trajectories, regulatory circuitry and interactions of cells, as well as provide a framework for understanding cellular dysregulation in human disease. Here we describe the idea, its potential utility, early proofs-of-concept, and some design considerations for the Human Cell Atlas.
]]></description>
<dc:creator>Regev, A.</dc:creator>
<dc:creator>Teichmann, S.</dc:creator>
<dc:creator>Lander, E. S.</dc:creator>
<dc:creator>Amit, I.</dc:creator>
<dc:creator>Benoist, C.</dc:creator>
<dc:creator>Birney, E.</dc:creator>
<dc:creator>Bodenmiller, B.</dc:creator>
<dc:creator>Campbell, P.</dc:creator>
<dc:creator>Carninci, P.</dc:creator>
<dc:creator>Clatworthy, M.</dc:creator>
<dc:creator>Clevers, H.</dc:creator>
<dc:creator>Deplancke, B.</dc:creator>
<dc:creator>Dunham, I.</dc:creator>
<dc:creator>Eberwine, J.</dc:creator>
<dc:creator>Eils, R.</dc:creator>
<dc:creator>Enard, W.</dc:creator>
<dc:creator>Farmer, A.</dc:creator>
<dc:creator>Fugger, L.</dc:creator>
<dc:creator>Gottgens, B.</dc:creator>
<dc:creator>Hacohen, N.</dc:creator>
<dc:creator>Haniffa, M.</dc:creator>
<dc:creator>Hemberg, M.</dc:creator>
<dc:creator>Kim, S. K.</dc:creator>
<dc:creator>Klenerman, P.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:creator>Lein, E.</dc:creator>
<dc:creator>Linnarsson, S.</dc:creator>
<dc:creator>Lundeberg, J.</dc:creator>
<dc:creator>Majumder, P.</dc:creator>
<dc:creator>Marioni, J.</dc:creator>
<dc:creator>Merad, M.</dc:creator>
<dc:creator>Mhlanga, M.</dc:creator>
<dc:creator>Nawijn, M.</dc:creator>
<dc:creator>Netea, M.</dc:creator>
<dc:creator>Nolan, G.</dc:creator>
<dc:creator>Pe'er, D.</dc:creator>
<dc:creator>Philipakis, A.</dc:creator>
<dc:creator>Ponting, C. P.</dc:creator>
<dc:creator>Quake, S. R.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Rozenblatt-Rosen, O.</dc:creator>
<dc:creator>Sanes, J. R.</dc:creator>
<dc:creator>Satija, R.</dc:creator>
<dc:creator>Shumacher, T.</dc:creator>
<dc:creator>Shalek, A. K</dc:creator>
<dc:date>2017-05-08</dc:date>
<dc:identifier>doi:10.1101/121202</dc:identifier>
<dc:title><![CDATA[The Human Cell Atlas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/125419v1?rss=1">
<title>
<![CDATA[
OGT binds a conserved C-terminal domain of TET1 to regulate TET1 activity and function in development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/125419v1?rss=1</link>
<description><![CDATA[
Mammalian TET enzymes oxidize 5-methylcytosine to 5-hydroxymethylcytosine and higher oxidized derivatives. TETs are targets of the enzyme OGT, which post-translationally modifies intracellular proteins in response to cellular nutrient status. The biological implications of the OGT-TET interaction have not been thoroughly explored. Here, we show for the first time that modification of TET1 by OGT enhances its activity in vitro. We identify a previously uncharacterized domain of TET1 responsible for binding to OGT and report a point mutation that disrupts the OGT-TET1 interaction. Finally, we show that the interaction between TET1 and OGT is necessary for TET1 to rescue tet mutant zebrafish hematopoetic stem cell formation, suggesting that OGT promotes TET1s function in development. Our results demonstrate regulation of TET activity by OGT in vitro and in vivo. These results link metabolism and epigenetic control, which may be relevant to the developmental and disease processes regulated by these two enzymes.
]]></description>
<dc:creator>Hrit, J.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Martin, E. A.</dc:creator>
<dc:creator>Goll, M.</dc:creator>
<dc:creator>Panning, B.</dc:creator>
<dc:date>2017-04-07</dc:date>
<dc:identifier>doi:10.1101/125419</dc:identifier>
<dc:title><![CDATA[OGT binds a conserved C-terminal domain of TET1 to regulate TET1 activity and function in development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/127043v1?rss=1">
<title>
<![CDATA[
Limited contribution of rare, noncoding variation to autism spectrum disorder from sequencing of 2,076 genomes in quartet families 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/127043v1?rss=1</link>
<description><![CDATA[
Genomic studies to date in autism spectrum disorder (ASD) have largely focused on newly arising mutations that disrupt protein coding sequence and strongly influence risk. We evaluate the contribution of noncoding regulatory variation across the size and frequency spectrum through whole genome sequencing of 519 ASD cases, their unaffected sibling controls, and parents. Cases carry a small excess of de novo (1.02-fold) noncoding variants, which is not significant after correcting for paternal age. Assessing 51,801 regulatory classes, no category is significantly associated with ASD after correction for multiple testing. The strongest signals are observed in coding regions, including structural variation not detected by previous technologies and missense variation. While rare noncoding variation likely contributes to risk in neurodevelopmental disorders, no category of variation has impact equivalent to loss-of-function mutations. Average effect sizes are likely to be smaller than that for coding variation, requiring substantially larger samples to quantify this risk.
]]></description>
<dc:creator>Werling, D. M.</dc:creator>
<dc:creator>Brand, H.</dc:creator>
<dc:creator>An, J.-Y.</dc:creator>
<dc:creator>Stone, M. R.</dc:creator>
<dc:creator>Glessner, J. T.</dc:creator>
<dc:creator>Zhu, L.</dc:creator>
<dc:creator>Collins, R. L.</dc:creator>
<dc:creator>Dong, S.</dc:creator>
<dc:creator>Layer, R. M.</dc:creator>
<dc:creator>Markenscoff-Papadimitriou, E.-C.</dc:creator>
<dc:creator>Farrell, A.</dc:creator>
<dc:creator>Schwartz, G. B.</dc:creator>
<dc:creator>Currall, B. B.</dc:creator>
<dc:creator>Dea, J.</dc:creator>
<dc:creator>Duhn, C.</dc:creator>
<dc:creator>Erdman, C.</dc:creator>
<dc:creator>Gilson, M.</dc:creator>
<dc:creator>Handsaker, R. E.</dc:creator>
<dc:creator>Kashin, S.</dc:creator>
<dc:creator>Klei, L.</dc:creator>
<dc:creator>Mandell, J. D.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Pochareddy, S.</dc:creator>
<dc:creator>Smith, L.</dc:creator>
<dc:creator>Walker, M. F.</dc:creator>
<dc:creator>Wang, H. Z.</dc:creator>
<dc:creator>Waterman, M. J.</dc:creator>
<dc:creator>He, X.</dc:creator>
<dc:creator>Kriegstein, A. R.</dc:creator>
<dc:creator>Rubenstein, J. L.</dc:creator>
<dc:creator>Sestan, N.</dc:creator>
<dc:creator>McCarroll, S. A.</dc:creator>
<dc:creator>Neale, B. M.</dc:creator>
<dc:creator>Coon, H.</dc:creator>
<dc:creator>Willsey, A. J.</dc:creator>
<dc:creator>Buxbaum, J. D.</dc:creator>
<dc:creator>Daly, M. J.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Quinlan, A.</dc:creator>
<dc:creator>Marth, G. T.</dc:creator>
<dc:creator>Roeder, K.</dc:creator>
<dc:creator>Devli</dc:creator>
<dc:date>2017-04-13</dc:date>
<dc:identifier>doi:10.1101/127043</dc:identifier>
<dc:title><![CDATA[Limited contribution of rare, noncoding variation to autism spectrum disorder from sequencing of 2,076 genomes in quartet families]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/127100v1?rss=1">
<title>
<![CDATA[
Nanopore detection of bacterial DNA base modifications 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/127100v1?rss=1</link>
<description><![CDATA[
The common bacterial base modification N6-methyladenine (m6A) is involved in many pathways related to an organisms ability to survive and interact with its environment. Recent research has shown that nanopore sequencing can detect m5C with per-read accuracy of upwards of 80% but m6A with significantly lower accuracy. Here we use a binary classifier to improve m6A classification by marking adenines as methylated or unmethylated based on differences between measured and expected current values as each adenine travels through the nanopore. We also illustrate the importance of read quality for base modification detection and compare to PacBio methylation calls. With recent demonstrations of nanopore sequencing in Antarctica and onboard the International Space Station, the ability to reliably characterize m6A presents an opportunity to further examine the role of methylation in bacterial adaptation to extreme or very remote environments.
]]></description>
<dc:creator>McIntyre, A. B. R.</dc:creator>
<dc:creator>Alexander, N.</dc:creator>
<dc:creator>Burton, A. S.</dc:creator>
<dc:creator>Castro-Wallace, S.</dc:creator>
<dc:creator>Chiu, C. Y.</dc:creator>
<dc:creator>John, K. K.</dc:creator>
<dc:creator>Stahl, S. E.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Mason, C. E.</dc:creator>
<dc:date>2017-04-13</dc:date>
<dc:identifier>doi:10.1101/127100</dc:identifier>
<dc:title><![CDATA[Nanopore detection of bacterial DNA base modifications]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/130484v1?rss=1">
<title>
<![CDATA[
Use antibiotics in cell culture with caution: genome-wide identification ofantibiotic-induced changes in gene expression and regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/130484v1?rss=1</link>
<description><![CDATA[
Standard cell culture guidelines often use media supplemented with antibiotics to prevent cell contamination. However, relatively little is known about the effect of antibiotic use in cell culture on gene expression and the extent to which this treatment could confound results. To comprehensively characterize the effect of antibiotic treatment on gene expression, we performed RNA-seq and ChIP-seq for H3K27ac on HepG2 cells, a human liver cell line commonly used for pharmacokinetic, metabolism and genomic studies, cultured in media supplemented with penicillin-streptomycin (PenStrep) or vehicle control. We identified 205 PenStrep-responsive genes, including transcription factors such as ATF3 that are likely to alter the regulation of other genes. Pathway analyses found a significant enrichment for "xenobiotic metabolism signaling" and "PXR/RXR activation" pathways. Our H3K27ac ChIP-seq identified 9,514 peaks that are PenStrep responsive. These peaks were enriched near genes that function in cell differentiation, tRNA modification, nuclease activity and protein dephosphorylation. Our results suggest that PenStrep treatment can significantly alter gene expression and regulation in a common liver cell type such as HepG2, advocating that antibiotic treatment should be taken into account when carrying out genetic, genomic or other biological assays in cultured cells.
]]></description>
<dc:creator>Ryu, A. H.</dc:creator>
<dc:creator>Eckalbar, W. L.</dc:creator>
<dc:creator>Kreimer, A.</dc:creator>
<dc:creator>Yosef, N.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2017-04-25</dc:date>
<dc:identifier>doi:10.1101/130484</dc:identifier>
<dc:title><![CDATA[Use antibiotics in cell culture with caution: genome-wide identification ofantibiotic-induced changes in gene expression and regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/131870v1?rss=1">
<title>
<![CDATA[
Topical Silver Diamine Fluoride For Dental Caries Arrest In Preschool Children: A Randomized Controlled Trial 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/131870v1?rss=1</link>
<description><![CDATA[
ObjectivesThe Stopping Cavities Trial investigated effectiveness and safety of 38% silver diamine fluoride in arresting caries lesions.nnMaterials and MethodsDouble-blind randomized placebo-controlled superiority trial with 2 parallel groups. Oregon preschools. 66 preschool children with [&ge;]1 lesion. 38% silver diamine fluoride or placebo (blue-tinted water), applied topically to the lesion. The primary endpoint was caries arrest (lesion inactivity, Nyvad criteria) 14-21 days post intervention. Dental plaque was collected from all children, and microbial composition was assessed by RNA sequencing from 2 lesions and 1 unaffected surface before treatment and at follow-up for 3 children from each group.nnResults and ConclusionMean fraction of arrested caries lesions in the silver diamine fluoride group was higher (0.72; 95% CI; 0.55, 0.84) than in the placebo group (0.05; 95% CI; 0.00, 0.16). Confirmatory analysis using generalized estimating equation log-linear regression, accounting for the number of treated surfaces and length of follow-up, indicated the fraction of arrested caries was significantly higher in the treatment group (relative risk, 17.3; 95% CI: 4.3 to 69.4). No harms were observed. RNA sequencing analysis identified no consistent changes in relative abundance of caries-associated microbes, nor emergence of antibiotic or metal resistance gene expression. Topical 38% silver diamine fluoride was effective and safe in arresting cavities in preschool children. The treatment is applicable to primary care practice and may reduce the burden of untreated tooth decay in the population.nnTrial RegistrationClinicalTrials.gov NCT02536040.nnClinical SignificanceIn this clinical trial, 72% of caries lesions were arrested by silver diamine fluoride, with no harms. Contrary to the presumed antibacterial mechanism, lesion bacterial composition changed negligibly. This simple topical treatment is applicable to primary care practice and may reduce the burden of untreated tooth decay in the population.
]]></description>
<dc:creator>Milgrom, P.</dc:creator>
<dc:creator>Horst, J. A.</dc:creator>
<dc:creator>Ludwig, S.</dc:creator>
<dc:creator>Rothen, M.</dc:creator>
<dc:creator>Chaffee, B. W.</dc:creator>
<dc:creator>Lyalina, S.</dc:creator>
<dc:creator>Pollard, K.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Mancl, L.</dc:creator>
<dc:date>2017-04-29</dc:date>
<dc:identifier>doi:10.1101/131870</dc:identifier>
<dc:title><![CDATA[Topical Silver Diamine Fluoride For Dental Caries Arrest In Preschool Children: A Randomized Controlled Trial]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/133132v1?rss=1">
<title>
<![CDATA[
Estimating Degree Of Polygenicity, Causal Effect Size Variance, And Confounding Bias In GWAS Summary Statistics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/133132v1?rss=1</link>
<description><![CDATA[
Estimating the polygenicity (proportion of causally associated single nucleotide polymorphisms (SNPs)) and discoverability (effect size variance) of causal SNPs for human traits is currently of considerable interest. SNP-heritability is proportional to the product of these quantities. We present a basic model, using detailed linkage disequilibrium structure from an extensive reference panel, to estimate these quantities from genome-wide association studies (GWAS) summary statistics. We apply the model to diverse phenotypes and validate the implementation with simulations. We find model polygenicities ranging from [~=] 2 x 10-5 to [~=] 4 x 10-3, with discoverabilities similarly ranging over two orders of magnitude. A power analysis allows us to estimate the proportions of phenotypic variance explained additively by causal SNPs reaching genome-wide significance at current sample sizes, and map out sample sizes required to explain larger portions of additive SNP heritability. The model also allows for estimating residual inflation (or deflation from over-correcting of z-scores), and assessing compatibility of replication and discovery GWAS summary statistics.

Author SummaryThere are ~10 million common variants in the genome of humans with European ancestry. For any particular phenotype a number of these variants will have some causal effect. It is of great interest to be able to quantify the number of these causal variants and the strength of their effect on the phenotype.

Genome wide association studies (GWAS) produce very noisy summary statistics for the association between subsets of common variants and phenotypes. For any phenotype, these statistics collectively are difficult to interpret, but buried within them is the true landscape of causal effects. In this work, we posit a probability distribution for the causal effects, and assess its validity using simulations. Using a detailed reference panel of ~11 million common variants - among which only a small fraction are likely to be causal, but allowing for non-causal variants to show an association with the phenotype due to correlation with causal variants - we implement an exact procedure for estimating the number of causal variants and their mean strength of association with the phenotype. We find that, across different phenotypes, both these quantities - whose product allows for lower bound estimates of heritability - vary by orders of magnitude.
]]></description>
<dc:creator>Holland, D.</dc:creator>
<dc:creator>Fan, C.-C.</dc:creator>
<dc:creator>Frei, O.</dc:creator>
<dc:creator>Shadrin, A. A.</dc:creator>
<dc:creator>Smeland, O. B.</dc:creator>
<dc:creator>Sundar, V. S.</dc:creator>
<dc:creator>ENIGMA,</dc:creator>
<dc:creator>Andreassen, O. A.</dc:creator>
<dc:creator>Dale, A. M.</dc:creator>
<dc:date>2017-05-24</dc:date>
<dc:identifier>doi:10.1101/133132</dc:identifier>
<dc:title><![CDATA[Estimating Degree Of Polygenicity, Causal Effect Size Variance, And Confounding Bias In GWAS Summary Statistics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/133967v1?rss=1">
<title>
<![CDATA[
Spike Discharge Prediction Based On Neuro-Fuzzy System 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/133967v1?rss=1</link>
<description><![CDATA[
This paper presents the development and evaluation of different versions of Neuro-Fuzzy model for prediction of spike discharge patterns. We aim to predict the spike discharge variation using first spike latency and frequency-following interval. In order to study the spike discharge dynamics, we analyzed the Cerebral Cortex data of the cat from [29]. Adaptive Neuro-Fuzzy Inference Systems (ANFIS), Wang and Mendel (WM), Dynamic evolving neural-fuzzy inference system (DENFIS), Hybrid neural Fuzzy Inference System (HyFIS), genetic for lateral tuning and rule selection of linguistic fuzzy system (GFS.LT.RS) and subtractive clustering and fuzzy c-means (SBC) algorithms are applied for data. Among these algorithms, ANFIS and GFS.LT.RS models have better performance. On the other hand, ANFIS and GFS.LT.RS algorithms can be used to predict the spike discharge dynamics as a function of first spike latency and frequency with a higher accuracy compared to other algorithms.
]]></description>
<dc:creator>Zarei, M.</dc:creator>
<dc:date>2017-05-03</dc:date>
<dc:identifier>doi:10.1101/133967</dc:identifier>
<dc:title><![CDATA[Spike Discharge Prediction Based On Neuro-Fuzzy System]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/136226v1?rss=1">
<title>
<![CDATA[
Ketogenic diet or BHB improves epileptiform spikes, memory, survival in Alzheimer’s model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/136226v1?rss=1</link>
<description><![CDATA[
Links between epilepsy and Alzheimers disease (AD) are seen in both human patients and mouse models. Human patients with AD may commonly have subclinical epileptiform spikes (EP spikes)1, and overt epilepsy is associated with more rapid cognitive decline2. Mechanistic studies in mouse models of Alzheimers disease (AD) have shown that altered network activity and epileptiform spikes stem from dysfunctional inhibitory interneurons3, which are key elements of cortical circuits underlying cognition4. Treatments that reduce epileptiform spikes improve cognition in these models5,6. Thus, targeting subclinical epileptiform activity may be a promising new therapeutic approach to AD7. Ketogenic diet (KD) has long been used to treat forms of epilepsy8, including Dravet syndrome, a childhood epilepsy caused by mutations in a gene that is critical for inhibitory interneuron function in mouse AD models5,9. However, the concurrent effects of a ketogenic diet on brain electrical activity, cognitive decline, and survival have not been tested, and the translational rationale and feasibility of such an intervention remain uncertain. Here we show that a ketogenic diet reduces epileptiform spikes in the hAPPJ20 mouse model of AD. Similar reduction of EP spikes is observed using a {beta}-hydroxybutyrate (BHB) ester in both AD and Dravet mice. A ketogenic diet improves context-dependent and visuo-spatial learning in hAPPJ20 mice. It also reduces the high seizure-related mortality observed in male mice of this model. Therapies derived from {beta}-hydroxybutyrate may have potential application in ameliorating cognitive dysfunction in AD through reducing subclinical epileptiform activity.
]]></description>
<dc:creator>Newman, J. C.</dc:creator>
<dc:creator>Kroll, F.</dc:creator>
<dc:creator>Ulrich, S.</dc:creator>
<dc:creator>Palop, J. J.</dc:creator>
<dc:creator>Verdin, E.</dc:creator>
<dc:date>2017-05-09</dc:date>
<dc:identifier>doi:10.1101/136226</dc:identifier>
<dc:title><![CDATA[Ketogenic diet or BHB improves epileptiform spikes, memory, survival in Alzheimer’s model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/136713v1?rss=1">
<title>
<![CDATA[
A Novel Reporter Allele For Monitoring Dll4 Expression Within The Embryonic And Adult Mouse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/136713v1?rss=1</link>
<description><![CDATA[
Canonical Notch signaling requires the presence of a membrane bound ligand and a corresponding transmembrane Notch receptor. Receptor engagement induces multiple proteolytic cleavage events culminating in the nuclear accumulation of the Notch intracellular domain and its binding to a transcriptional co-factor to mediate gene expression. Notch signaling networks are essential regulators of vascular patterning and angiogenesis, as well as myriad other biological processes. Delta-like 4 (Dll4) encodes the earliest Notch ligand detected in arterial cells, and is enriched in sprouting endothelial tip cells. Dll4 expression has often been inferred by proxy using a lacZ knockin reporter allele. This is problematic, as a single copy of Dll4 is haploinsufficient. Additionally, Notch activity regulates Dll4 transcription, making it unclear whether these reporter lines accurately reflect Dll4 expression. Accordingly, accurately defining Dll4 expression is essential for determining its role in development and disease. To address these limitations, we generated a novel BAC transgenic allele with a nuclear-localized {beta}-galactosidase reporter (Dll4-BAC-nlacZ). Through a comparative analysis, we show the BAC line overcomes previous issues of haploinsufficiency, it recapitulates Dll4 expression in vivo, and allows superior visualization and imaging. As such, this novel Dll4 reporter is an important addition to the growing Notch toolkit.nnSummary Statement:nnWe have developed a novel reporter line, free from complicating factors associated with previous alleles, for monitoring Dll4 expression, at a cellular resolution, in the developing and adult mouse.
]]></description>
<dc:creator>Herman, A. M.</dc:creator>
<dc:creator>Rhyner, A. M.</dc:creator>
<dc:creator>Devine, W. P.</dc:creator>
<dc:creator>Marrelli, S. P.</dc:creator>
<dc:creator>Bruneau, B. G.</dc:creator>
<dc:creator>Wythe, J. D.</dc:creator>
<dc:date>2017-05-11</dc:date>
<dc:identifier>doi:10.1101/136713</dc:identifier>
<dc:title><![CDATA[A Novel Reporter Allele For Monitoring Dll4 Expression Within The Embryonic And Adult Mouse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/139352v1?rss=1">
<title>
<![CDATA[
Extending Chemical Perturbations Of The Ubiquitin Fitness Landscape In A Classroom Setting 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/139352v1?rss=1</link>
<description><![CDATA[
Although the primary protein sequence of ubiquitin (Ub) is extremely stable over evolutionary time, it is highly tolerant to mutation during selection experiments performed in the laboratory. We have proposed that this discrepancy results from the difference between fitness under laboratory culture conditions and the selective pressures in changing environments over evolutionary time scales. Building on our previous work (Mavor et al 2016), we used deep mutational scanning to determine how twelve new chemicals (3-Amino-1,2,4-triazole, 5-fluorocytosine, Amphotericin B, CaCl2, Cerulenin, Cobalt Acetate, Menadione, Nickel Chloride, p-fluorophenylalanine, Rapamycin, Tamoxifen, and Tunicamycin) reveal novel mutational sensitivities of ubiquitin residues. We found sensitization of Lys63 in eight new conditions. In total, our experiments have uncovered a sensitizing condition for every position in Ub except Ser57 and Gln62. By determining the Ubiquitin fitness landscape under different chemical constraints, our work helps to resolve the inconsistencies between deep mutational scanning experiments and sequence conservation over evolutionary timescales.nnBuilds onMavor D, Barlow KA, Thompson S, Barad BA, Bonny AR, Cario CL, Gaskins G, Liu Z, Deming L, Axen SD, Caceres E, Chen W, Cuesta A, Gate R, Green EM, Hulce KR, Ji W, Kenner LR, Mensa B, Morinishi LS, Moss SM, Mravic M, Muir RK, Niekamp S, Nnadi CI, Palovcak E, Poss EM, Ross TD, Salcedo E, See S, Subramaniam M, Wong AW, Li J, Thorn KS, Conchuir SO, Roscoe BP, Chow ED, DeRisi JL, Kortemme T, Bolon DN, Fraser JS. Determination of Ubiquitin Fitness Landscapes Under Different Chemical Stresses in a Classroom Setting. eLife. 2016.nnImpact StatementWe organized a project-based course that used deep mutational scanning in multiple chemical conditions to resolve the inconsistencies between tolerance to mutations in laboratory conditions and sequence conservation over evolutionary timescales.
]]></description>
<dc:creator>Mavor, D.</dc:creator>
<dc:creator>Barlow, K.</dc:creator>
<dc:creator>Asarnow, D.</dc:creator>
<dc:creator>Birman, Y.</dc:creator>
<dc:creator>Britain, D.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Green, E. M.</dc:creator>
<dc:creator>Kenner, L. R.</dc:creator>
<dc:creator>Mensa, B.</dc:creator>
<dc:creator>Morinishi, L. S.</dc:creator>
<dc:creator>Nelson, C. A.</dc:creator>
<dc:creator>Poss, E. M.</dc:creator>
<dc:creator>Suresh, P.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Arhar, T.</dc:creator>
<dc:creator>Ary, B. E.</dc:creator>
<dc:creator>Bauer, D. P.</dc:creator>
<dc:creator>Bergman, I. D.</dc:creator>
<dc:creator>Brunetti, R. M.</dc:creator>
<dc:creator>Chio, C. M.</dc:creator>
<dc:creator>Dai, S. A.</dc:creator>
<dc:creator>Dickinson, M. S.</dc:creator>
<dc:creator>Elledge, S.</dc:creator>
<dc:creator>Hendel, N. L.</dc:creator>
<dc:creator>Helsell, C. V. M.</dc:creator>
<dc:creator>Kang, E.</dc:creator>
<dc:creator>Kern, N.</dc:creator>
<dc:creator>Khoroshkin, M. S.</dc:creator>
<dc:creator>Kirkemo, L. L.</dc:creator>
<dc:creator>Lewis, G. R.</dc:creator>
<dc:creator>Lou, K.</dc:creator>
<dc:creator>Marin, W. M.</dc:creator>
<dc:creator>Maxwell, A. M.</dc:creator>
<dc:creator>McTigue, P. F.</dc:creator>
<dc:creator>Meyers-Turnbull, D.</dc:creator>
<dc:creator>Nagy, T. L.</dc:creator>
<dc:creator>Natale, A. M.</dc:creator>
<dc:creator>Oltion, K.</dc:creator>
<dc:creator>Pourmal, S.</dc:creator>
<dc:creator>Reder, G. K.</dc:creator>
<dc:creator>Rettko, N. J.</dc:creator>
<dc:creator>Rohweder, P. J.</dc:creator>
<dc:creator>Schwarz, D. M. C.</dc:creator>
<dc:date>2017-05-17</dc:date>
<dc:identifier>doi:10.1101/139352</dc:identifier>
<dc:title><![CDATA[Extending Chemical Perturbations Of The Ubiquitin Fitness Landscape In A Classroom Setting]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/140426v1?rss=1">
<title>
<![CDATA[
Promoter Or Enhancer Activation By CRISPRa Rescues Haploinsufficiency Caused Obesity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/140426v1?rss=1</link>
<description><![CDATA[
Haploinsufficiency, having only one functional copy of a gene, leads to a wide range of human disease and has been associated with over 300 genes. Here, we tested whether CRISPR activation (CRISPRa) could rescue a haploinsufficient disease in vivo. Haploinsufficiency of Sim1, a transcription factor involved in the leptin pathway, results in severe obesity in humans and mice. CRISPRa targeting of either the Sim1 promoter or its ~270kb distant hypothalamic enhancer using transgenic mice, rescued the obesity phenotype in Sim1 heterozygous mice. Interestingly, despite using a ubiquitous promoter for CRISPRa, Sim1 was upregulated only in tissues where the promoter or enhancer are active, suggesting that cis-regulatory elements can determine CRISPRa tissue-specificity. To further relate this to therapy, we injected CRISPRa adeno associated virus into the hypothalamus, leading to reversal of the obesity phenotype. This therapeutic strategy could be used to rescue numerous diseases resulting from altered gene dosage.
]]></description>
<dc:creator>Matharu, N.</dc:creator>
<dc:creator>Rattanasopha, S.</dc:creator>
<dc:creator>Maliskova, L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Hardin, A.</dc:creator>
<dc:creator>Vaisse, C.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2017-05-22</dc:date>
<dc:identifier>doi:10.1101/140426</dc:identifier>
<dc:title><![CDATA[Promoter Or Enhancer Activation By CRISPRa Rescues Haploinsufficiency Caused Obesity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/143362v1?rss=1">
<title>
<![CDATA[
Senescence As A Defense Strategy Against Parasites 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/143362v1?rss=1</link>
<description><![CDATA[
The teleology of aging has been one of the more vexing and controversial question in biology. One potential evolutionary driver of programmed aging is selection pressure from parasites and other infectious organisms. While selection pressure from parasites and other infectious organisms have long been considered by many biologists to have led to the evolution of sexual reproduction, it has only rarely been considered as a potential driver for evolution of aging, a biological process that likely evolved contemporaneously with sexual reproduction. Here I describe stochastic simulations of host and parasite populations with senescence as an independent variable. The results show that populations with more rapid senescence bear lower parasite loads and oscillate more quickly through alternate phenotypes with differential resistance against parasites. I conclude that programmed aging and death may promote host evasion of parasites in a co-evolutionary competition against parasites.
]]></description>
<dc:creator>Chin, R. Y.</dc:creator>
<dc:date>2017-05-29</dc:date>
<dc:identifier>doi:10.1101/143362</dc:identifier>
<dc:title><![CDATA[Senescence As A Defense Strategy Against Parasites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/143719v1?rss=1">
<title>
<![CDATA[
Interphase-Arrested Drosophila Embryos Initiate Mid-Blastula Transition At A Low Nuclear-Cytoplasmic Ratio 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/143719v1?rss=1</link>
<description><![CDATA[
Externally deposited eggs begin development with an immense cytoplasm and a single overwhelmed nucleus. Rapid mitotic cycles restore normality as the ratio of nuclei to cytoplasm (N/C) increases. At the 14th cell cycle in Drosophila embryos, the cell cycle slows, transcription increases, and morphogenesis begins at the Mid-Blastula Transition (MBT). To explore the role of N/C in MBT timing, we blocked N/C-increase by downregulating cyclin/Cdk1 to arrest early cell cycles. Embryos arrested in cell cycle 12 cellularized, initiated gastrulation movements and activated transcription of genes previously described as N/C dependent. Thus, occurrence of these events is not directly coupled to N/C-increase. However, N/C might act indirectly. Increasing N/C promotes cyclin/Cdk1 downregulation which otherwise inhibits many MBT events. By experimentally inducing downregulation of cyclin/Cdk1, we bypassed this input of N/C-increase. We describe a regulatory cascade wherein the increasing N/C downregulates cyclin/Cdk1 to promote increasing transcription and the MBT.nnImpact statementBy showing that cell-cycle arrest allows early Drosophila embryos to progress to later stages, this work eliminates numerous models for embryonic timing and shows the dominating influence of cell-cycle slowing.
]]></description>
<dc:creator>Strong, I.</dc:creator>
<dc:creator>Yuan, K.</dc:creator>
<dc:creator>O'Farrell, P. H.</dc:creator>
<dc:date>2017-06-01</dc:date>
<dc:identifier>doi:10.1101/143719</dc:identifier>
<dc:title><![CDATA[Interphase-Arrested Drosophila Embryos Initiate Mid-Blastula Transition At A Low Nuclear-Cytoplasmic Ratio]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/144113v1?rss=1">
<title>
<![CDATA[
Characterization Of Vif Domains That Mediate Feline Immunodeficiency Virus Antagonism Of APOBEC3-H And APOBEC3-CH Restriction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/144113v1?rss=1</link>
<description><![CDATA[
Feline immunodeficiency virus (FIV) Vif mediates degradation of two anti-lentiviral feline APOBEC3 (fA3) proteins, fA3Z3 and fA3Z2bZ3. HIV-1 Vif targets the restriction factor human APOBEC3G (A3G, hA3Z2g-Z1c) for proteasome degradation to mediate viral evasion. Despite this similarity, FIV and HIV-1 Vif share limited homology. Vif binds hA3Z2g-Z1c through its N-terminal region, while its C-terminal region binds to an E3-ligase complex containing Cullin5 and Elongin B/C. Further, HIV-1 Vif contains critical domains in its C-terminus, including an adjacent BC box, the only shared domain between FIV and HIV-1 Vif, and a non-classical zinc finger (HCCH) domain. Felid lentivirus Vif, however, contains a highly conserved KCCC motif. While both Vifs have evolved to counteract select A3 antiretroviral proteins, the FIV Vif domains necessary to target fA3s for degradation are incompletely understood. To identify these domains, we used the well-characterized HIV-1 Vif domains to show that distinct mutations within the BC box of FIV Vif prevent fA3Z3 and fA3Z2bZ3 degradation and reduce virion infectivity. We also found that mutating any single residue in the KCCC motif blocked fA3 targeting and impaired FIV infectivity and replication. These mutations also failed to disrupt the FIV Vif and Cullin5 interaction. Further, we showed that, in contrast to the HCCH domain in HIV-1 Vif, the KCCC domain of FIV Vif does not bind zinc. However, unlike HIV-1 Vif, FIV Vif (C36 isolate) reduces intracellular levels of co-expressed Cullin5 proteins, a novel finding. Our results reveal important C-terminal residues in FIV Vif and show that the BC box and KCCC regions are critical for fA3 degradation, infectivity, and spreading replication.
]]></description>
<dc:creator>Sims, O. L.</dc:creator>
<dc:creator>Maynard, E.</dc:creator>
<dc:creator>Poeschla, E. M.</dc:creator>
<dc:date>2017-05-30</dc:date>
<dc:identifier>doi:10.1101/144113</dc:identifier>
<dc:title><![CDATA[Characterization Of Vif Domains That Mediate Feline Immunodeficiency Virus Antagonism Of APOBEC3-H And APOBEC3-CH Restriction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/146811v1?rss=1">
<title>
<![CDATA[
An integrated genomic analysis of anaplastic meningioma identifies prognostic molecular signatures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/146811v1?rss=1</link>
<description><![CDATA[
Anaplastic meningioma is a rare and aggressive brain tumor characterised by intractable recurrences and dismal outcomes. Here, we present an integrated analysis of the whole genome, transcriptome and methylation profiles of primary and recurrent anaplastic meningioma. A key finding was the delineation of two distinct molecular subgroups that were associated with diametrically opposed survival outcomes. Relative to lower grade meningiomas, anaplastic tumors harbored frequent driver mutations in SWI/SNF complex genes, which were confined to the poor prognosis subgroup. Our analyses discern two biologically distinct variants of anaplastic meningioma with potential prognostic and therapeutic significance.
]]></description>
<dc:creator>Collord, G.</dc:creator>
<dc:creator>Tarpey, P.</dc:creator>
<dc:creator>Kurbatova, N.</dc:creator>
<dc:creator>Martincorena, I.</dc:creator>
<dc:creator>Moran, S.</dc:creator>
<dc:creator>Castro, M.</dc:creator>
<dc:creator>Nagy, T.</dc:creator>
<dc:creator>Bignell, G.</dc:creator>
<dc:creator>Maura, F.</dc:creator>
<dc:creator>Berna, J.</dc:creator>
<dc:creator>Tubio, J. M.</dc:creator>
<dc:creator>McMurran, C. E.</dc:creator>
<dc:creator>Young, A. M. H.</dc:creator>
<dc:creator>Young, M. D.</dc:creator>
<dc:creator>Noorani, I.</dc:creator>
<dc:creator>Price, S. J.</dc:creator>
<dc:creator>Watts, C.</dc:creator>
<dc:creator>Leipnitz, E.</dc:creator>
<dc:creator>Kirsch, M.</dc:creator>
<dc:creator>Schackert, G.</dc:creator>
<dc:creator>Pearson, D.</dc:creator>
<dc:creator>Devadass, A.</dc:creator>
<dc:creator>Ram, Z.</dc:creator>
<dc:creator>Collins, V. P.</dc:creator>
<dc:creator>Allinson, K.</dc:creator>
<dc:creator>Jenkinson, M.</dc:creator>
<dc:creator>Zakaria, R.</dc:creator>
<dc:creator>Syed, K.</dc:creator>
<dc:creator>Hanemann, C. O.</dc:creator>
<dc:creator>Dunn, J.</dc:creator>
<dc:creator>McDermott, M.</dc:creator>
<dc:creator>Kirollos, R.</dc:creator>
<dc:creator>Vassiliou, G. S.</dc:creator>
<dc:creator>Esteller, M.</dc:creator>
<dc:creator>Behjati, S.</dc:creator>
<dc:creator>Brazma, A.</dc:creator>
<dc:creator>Santarius, T.</dc:creator>
<dc:creator>McDermott, U.</dc:creator>
<dc:date>2017-06-06</dc:date>
<dc:identifier>doi:10.1101/146811</dc:identifier>
<dc:title><![CDATA[An integrated genomic analysis of anaplastic meningioma identifies prognostic molecular signatures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/147207v1?rss=1">
<title>
<![CDATA[
Placental gene expression mediates the interaction between obstetrical history and genetic risk for schizophrenia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/147207v1?rss=1</link>
<description><![CDATA[
Defining the environmental context in which genes enhance susceptibility can provide insight into the pathogenesis of complex disorders, like schizophrenia. Here we show that the intrauterine and perinatal environment modulates the association of schizophrenia with genomic risk, as measured with polygenic risk scores (PRS) based primarily on GWAS significant variants. Genomic risk interacts with intrauterine and perinatal complications (Early Life Complications, ELCs) in each of three independent samples from USA, Italy and Germany (overall n= 1693, p= 6e-05). In each sample, the liability of schizophrenia explained by PRS is nominally more than five times greater in the presence of a history of ELCs compared with its absence. In each sample, patients with positive ELC histories have higher PRS than patients without ELCs, which is further confirmed in two additional patient samples from Germany and Japan (overall n=2038, p= 1e-04). The gene set based on the schizophrenia loci interacting with ELCs is highly expressed in multiple placental compartments and dynamically regulated in placenta from complicated in comparison with normal pregnancies. The same genes are differentially up-regulated in placentae from male compared with female offspring. The interaction between genomic risk and ELCs is mainly driven by GWAS significant loci enriched for genes highly expressed in the various placenta samples. Molecular pathway analyses based on the genes not driving this interaction reflect previous analyses about schizophrenia risk-genes, while genes highly and differentially expressed in placentae implicate an orthogonal biology involving cellular stress. These results suggest that the most significant genetic variants detected by current schizophrenia GWAS contribute to risk in part by converging on a developmental trajectory sensitive to events affecting placentation, which may underlie the male preponderance of schizophrenia and offer new insights into primary prevention.
]]></description>
<dc:creator>Ursini, G.</dc:creator>
<dc:creator>Punzi, G.</dc:creator>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>Marenco, S.</dc:creator>
<dc:creator>Robinson, J.</dc:creator>
<dc:creator>Porcelli, A.</dc:creator>
<dc:creator>Hamilton, E.</dc:creator>
<dc:creator>Mitjans, M.</dc:creator>
<dc:creator>Maddalena, G.</dc:creator>
<dc:creator>Begemann, M.</dc:creator>
<dc:creator>Seidel, J.</dc:creator>
<dc:creator>Yanamori, H.</dc:creator>
<dc:creator>Jaffe, A. E.</dc:creator>
<dc:creator>Berman, K. F.</dc:creator>
<dc:creator>Egan, M. F.</dc:creator>
<dc:creator>Straub, R. E.</dc:creator>
<dc:creator>Colantuoni, C.</dc:creator>
<dc:creator>Blasi, G.</dc:creator>
<dc:creator>Hashimoto, R.</dc:creator>
<dc:creator>Rujescu, D.</dc:creator>
<dc:creator>Ehrenreich, H.</dc:creator>
<dc:creator>Bertolino, A.</dc:creator>
<dc:creator>Weinberger, D. R.</dc:creator>
<dc:date>2017-06-07</dc:date>
<dc:identifier>doi:10.1101/147207</dc:identifier>
<dc:title><![CDATA[Placental gene expression mediates the interaction between obstetrical history and genetic risk for schizophrenia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/149385v1?rss=1">
<title>
<![CDATA[
Epigenetic maintenance of DNA methylation after evolutionary loss of the de novo methyltransferase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/149385v1?rss=1</link>
<description><![CDATA[
Cytosine methylation of DNA is a widespread modification of DNA that plays numerous critical roles, yet has been lost many times in diverse eukaryotic lineages. In the yeast Cryptococcus neoformans, CG methylation occurs in transposon-rich repeats and requires the DNA methyltransferase, Dnmt5. We show that Dnmt5 displays exquisite maintenance-type specificity in vitro and in vivo and utilizes similar in vivo cofactors as the metazoan maintenance methylase Dnmt1. Remarkably, phylogenetic and functional analysis revealed that the ancestral species lost the gene for a de novo methylase, DnmtX, between 50-150 MYA. We examined how methylation has persisted since the ancient loss of DnmtX. Experimental and comparative studies reveal efficient replication of methylation patterns in C. neoformans, rare stochastic methylation loss and gain events, and the action of natural selection. We propose that an epigenome has been propagated for >50 MY through a process analogous to Darwinian evolution of the genome.
]]></description>
<dc:creator>Catania, S.</dc:creator>
<dc:creator>Dumesic, P. A.</dc:creator>
<dc:creator>Stoddard, C.</dc:creator>
<dc:creator>Cooke, S.</dc:creator>
<dc:creator>Burke, J.</dc:creator>
<dc:creator>Cuomo, C. A.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:date>2017-06-13</dc:date>
<dc:identifier>doi:10.1101/149385</dc:identifier>
<dc:title><![CDATA[Epigenetic maintenance of DNA methylation after evolutionary loss of the de novo methyltransferase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/156760v1?rss=1">
<title>
<![CDATA[
Diffusion as a ruler: modeling kinesin diffusion as a length sensor for intraflagellar transport 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/156760v1?rss=1</link>
<description><![CDATA[
An important question in cell biology is whether cells are able to measure size, either whole cell size or organelle size. Perhaps cells have an internal chemical representation of size that can be used to precisely regulate growth, or perhaps size is just an accident that emerges due to constraint of nutrients. The eukaryotic flagellum is an ideal model for studying size sensing and control because its linear geometry makes it essentially one-dimensional, greatly simplifying mathematical modeling. The assembly of flagella is regulated by intraflagellar transport (IFT), in which kinesin motors carry cargo adaptors for flagellar proteins along the flagellum and then deposit them at the tip, lengthening the flagellum. The rate at which IFT motors are recruited to begin transport into the flagellum is anticorrelated with the flagellar length, implying some kind of communication between the base and the tip and possibly indicating that cells contain some mechanism for measuring flagellar length. Although it is possible to imagine many complex scenarios in which additional signaling molecules sense length and carry feedback signals to the cell body to control IFT, might the already-known components of the IFT system be sufficient to allow length dependence of IFT? Here, we investigate a model in which the anterograde kinesin motors unbind after cargo delivery, diffuse back to the base, and are subsequently reused to power entry of new IFT trains into the flagellum. By modeling such a system at three different levels of abstraction we are able to show that the diffusion time of the motors can in principle be sufficient to serve as a proxy for length measurement. In all three implementations, we found that the diffusion model can not only achieve a stable steady-state length without the addition of any other signaling molecules or pathways, but also is able to produce the anticorrelation between length and IFT recruitment rate that has been observed in quantitative imaging studies.
]]></description>
<dc:creator>Hendel, N. L.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2017-06-27</dc:date>
<dc:identifier>doi:10.1101/156760</dc:identifier>
<dc:title><![CDATA[Diffusion as a ruler: modeling kinesin diffusion as a length sensor for intraflagellar transport]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/156893v1?rss=1">
<title>
<![CDATA[
A Bulky Glycocalyx Fosters Metastasis Formation by Promoting G1 Cell Cycle Progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/156893v1?rss=1</link>
<description><![CDATA[
Metastasis depends upon cancer cell growth and survival within the metastatic niche. Tumors which remodel their glycocalyces, by overexpressing bulky glycoproteins like mucins, exhibit a higher predisposition to metastasize, but the role of mucins in oncogenesis remains poorly understood. Here we report that a bulky glycocalyx promotes the expansion of disseminated tumor cells in vivo by fostering integrin adhesion assembly to permit G1 cell cycle progression. We engineered tumor cells to display glycocalyces of various thicknesses by coating them with synthetic mucin-mimetic glycopolymers. Cells adorned with longer glycopolymers showed increased metastatic potential, enhanced cell cycle progression, and greater levels of integrin-FAK mechanosignaling and Akt signaling in a syngeneic mouse model of metastasis. These effects were mirrored by expression of the ectodomain of cancer-associated mucin MUC1. These findings functionally link mucinous proteins with tumor aggression, and offer a new view of the cancer glycocalyx as a major driver of disease progression.
]]></description>
<dc:creator>Woods, E. C.</dc:creator>
<dc:creator>Barnes, J. M.</dc:creator>
<dc:creator>Pickup, M. W.</dc:creator>
<dc:creator>Pedram, K.</dc:creator>
<dc:creator>Kai, F.</dc:creator>
<dc:creator>Hollander, M. J.</dc:creator>
<dc:creator>Weaver, V. M.</dc:creator>
<dc:creator>Bertozzi, C. R.</dc:creator>
<dc:date>2017-06-28</dc:date>
<dc:identifier>doi:10.1101/156893</dc:identifier>
<dc:title><![CDATA[A Bulky Glycocalyx Fosters Metastasis Formation by Promoting G1 Cell Cycle Progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/157198v1?rss=1">
<title>
<![CDATA[
A Genome-wide Association and Admixture Mapping Study of Bronchodilator Drug Response in African Americans with Asthma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/157198v1?rss=1</link>
<description><![CDATA[
BackgroundShort-acting B2-adrenergic receptor agonists (SABAs) are the most commonly prescribed asthma medications worldwide. Response to SABAs is measured as bronchodilator drug response (BDR), which varies among racial/ethnic groups in the U.S 1, 2. However, the genetic variation that contributes to BDR is largely undefined in African Americans with asthma3nnObjectiveTo identify genetic variants that may contribute to differences in BDR in African Americans with asthma.nnMethodsWe performed a genome-wide association study of BDR in 949 African American children with asthma, genotyped with the Axiom World Array 4 (Affymetrix, Santa Clara, CA) followed by imputation using 1000 Genomes phase 3 genotypes. We used linear regression models adjusting for age, sex, body mass index and genetic ancestry to test for an association between BDR and genotype at single nucleotide polymorphisms (SNPs). To increase power and distinguish between shared vs. population-specific associations with BDR in children with asthma, we performed a meta-analysis across 949 African Americans and 1,830 Latinos (Total=2,779). Lastly, we performed genome-wide admixture mapping to identify regions whereby local African or European ancestry is associated with BDR in African Americans. Two additional populations of 416 Latinos and 1,325 African Americans were used to replicate significant associations.nnResultsWe identified a population-specific association with an intergenic SNP on chromosome 9q21 that was significantly associated with BDR (rs73650726, p=7.69 x 10-9). A trans-ethnic meta-analysis across African Americans and Latinos identified three additional SNPs within the intron of PRKG1 that were significantly associated with BDR (rs7903366, rs7070958, and rs7081864, p[&le;]5 x 10-8).nnConclusionsOur findings indicate that both population specific and shared genetic variation contributes to differences in BDR in minority children with asthma, and that the genetic underpinnings of BDR may differ between racial/ethnic groups.nnKey messagesO_LIA GWAS for BDR in African American children with asthma identified an intergenic population specific variant at 9q21 to be associated with increased bronchodilator drug response (BDR).nC_LIO_LIA meta-analysis of GWAS across African Americans and Latinos identified shared genetic variants at 10q21 in the intron of PRKG1 to be associated with differences in BDR.nC_LIO_LIFurther genetic studies need to be performed in diverse populations to identify the full set of genetic variants that contribute to BDR.nC_LI
]]></description>
<dc:creator>Spear, M. L.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Pino-Yanes, M.</dc:creator>
<dc:creator>Huntsman, S.</dc:creator>
<dc:creator>Eng, C.</dc:creator>
<dc:creator>Levin, A. M.</dc:creator>
<dc:creator>White, M. J.</dc:creator>
<dc:creator>McGarry, M. E.</dc:creator>
<dc:creator>Thakur, N.</dc:creator>
<dc:creator>Galanter, J. M.</dc:creator>
<dc:creator>Mak, A. C. Y.</dc:creator>
<dc:creator>Oh, S. S.</dc:creator>
<dc:creator>Davis, A.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Farber, H. J.</dc:creator>
<dc:creator>Meade, K.</dc:creator>
<dc:creator>Avila, P. C.</dc:creator>
<dc:creator>Serebrisky, D.</dc:creator>
<dc:creator>Lenoir, M. A.</dc:creator>
<dc:creator>Brigino-Buenaventura, E. A.</dc:creator>
<dc:creator>Rodriquez Cintron, W.</dc:creator>
<dc:creator>Thyne, S. M.</dc:creator>
<dc:creator>Rodriguez-Santana, J. R.</dc:creator>
<dc:creator>Ford, J. G.</dc:creator>
<dc:creator>Chapela, R.</dc:creator>
<dc:creator>Moreno Estrada, A.</dc:creator>
<dc:creator>Sandoval, K.</dc:creator>
<dc:creator>Seibold, M. A.</dc:creator>
<dc:creator>Williams, L. K.</dc:creator>
<dc:creator>Winkler, C. A.</dc:creator>
<dc:creator>Torgerson, D. G.</dc:creator>
<dc:creator>Hernandez, R. D.</dc:creator>
<dc:creator>Burchard, E. G.</dc:creator>
<dc:date>2017-06-28</dc:date>
<dc:identifier>doi:10.1101/157198</dc:identifier>
<dc:title><![CDATA[A Genome-wide Association and Admixture Mapping Study of Bronchodilator Drug Response in African Americans with Asthma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/160440v1?rss=1">
<title>
<![CDATA[
Toward leveraging big data in human functional connectomics: Generalization of brain graphs across scanners, sessions, and paradigms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/160440v1?rss=1</link>
<description><![CDATA[
While graph theoretical modeling has dramatically advanced our understanding of complex brain systems, the feasibility of aggregating brain graphic data in large imaging consortia remains unclear. Here, using a battery of cognitive, emotional and resting fMRI paradigms, we investigated the reproducibility of functional connectomic measures across multiple sites and sessions. Our results revealed overall fair to excellent reliability for a majority of measures during both rest and tasks, in particular for those quantifying connectivity strength, network segregation and network integration. Higher reliabilities were detected for cognitive tasks (vs rest) and for weighted networks (vs binary networks). While network diagnostics for several primary functional systems were consistently reliable independently of paradigm, those for cognitive-emotional systems were reliable predominantly when challenged by task. Different data aggregation approaches yielded significantly different reliability. In addition, we showed that after accounting for observed reliability, satisfactory statistical power can be achieved in the multisite context with a total sample size of approximately 250 when the effect size is at least moderate. Our findings provide direct evidence for the generalizability of brain graphs for both resting and task paradigms in large consortia and encourage the use of multisite, multisession scans to enhance power for human functional connectomic studies.
]]></description>
<dc:creator>Cao, H.</dc:creator>
<dc:creator>McEwen, S.</dc:creator>
<dc:creator>Bearden, C.</dc:creator>
<dc:creator>Addington, J.</dc:creator>
<dc:creator>Goodyear, B.</dc:creator>
<dc:creator>Cadenhead, K.</dc:creator>
<dc:creator>Mirzakhanian, H.</dc:creator>
<dc:creator>Cornblatt, B.</dc:creator>
<dc:creator>Olvet, D.</dc:creator>
<dc:creator>Mathalon, D.</dc:creator>
<dc:creator>McGlashan, T.</dc:creator>
<dc:creator>Perkins, D.</dc:creator>
<dc:creator>Belger, A.</dc:creator>
<dc:creator>Seidman, L.</dc:creator>
<dc:creator>Thermenos, H.</dc:creator>
<dc:creator>Tsuang, M.</dc:creator>
<dc:creator>van Erp, T.</dc:creator>
<dc:creator>Walker, E.</dc:creator>
<dc:creator>Hamann, S.</dc:creator>
<dc:creator>Woods, S.</dc:creator>
<dc:creator>Cannon, T.</dc:creator>
<dc:date>2017-07-07</dc:date>
<dc:identifier>doi:10.1101/160440</dc:identifier>
<dc:title><![CDATA[Toward leveraging big data in human functional connectomics: Generalization of brain graphs across scanners, sessions, and paradigms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/160499v1?rss=1">
<title>
<![CDATA[
Whole Genome Sequencing in Psychiatric Disorders: the WGSPD Consortium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/160499v1?rss=1</link>
<description><![CDATA[
As technology advances, whole genome sequencing (WGS) is likely to supersede other genotyping technologies. The rate of this change depends on its relative cost and utility. Variants identified uniquely through WGS may reveal novel biological pathways underlying complex disorders and provide high-resolution insight into when, where, and in which cell type these pathways are affected. Alternatively, cheaper and less computationally intensive approaches may yield equivalent insights. Understanding the role of rare variants in the noncoding gene-regulating genome, through pilot WGS projects, will be critical to determine which of these two extremes best represents reality. With large cohorts, well-defined risk loci, and a compelling need to understand the underlying biology, psychiatric disorders have a role to play in this preliminary WGS assessment. The WGSPD consortium will integrate data for 18,000 individuals with psychiatric disorders, beginning with autism spectrum disorder, schizophrenia, bipolar disorder, and major depressive disorder, along with over 150,000 controls.
]]></description>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Neale, B. M.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Werling, D. M.</dc:creator>
<dc:creator>An, J.-Y.</dc:creator>
<dc:creator>Dong, S.</dc:creator>
<dc:creator>- Whole Genome Sequencing for Psychiatric Disorders,</dc:creator>
<dc:creator>Abecasis, G.</dc:creator>
<dc:creator>Arguello, P. A.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Boehnke, M.</dc:creator>
<dc:creator>Daly, M.</dc:creator>
<dc:creator>Eggan, K.</dc:creator>
<dc:creator>Geschwind, D. H.</dc:creator>
<dc:creator>Glahn, D.</dc:creator>
<dc:creator>Goldstein, D. B.</dc:creator>
<dc:creator>Gur, R. E.</dc:creator>
<dc:creator>Handsaker, R. E.</dc:creator>
<dc:creator>McCarroll, S. A.</dc:creator>
<dc:creator>Ophoff, R. A.</dc:creator>
<dc:creator>Palotie, A.</dc:creator>
<dc:creator>Pato, C.</dc:creator>
<dc:creator>Sabatti, C.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Willsey, A. J.</dc:creator>
<dc:creator>Hyman, S. E.</dc:creator>
<dc:creator>Addington, A.</dc:creator>
<dc:creator>Lehner, T.</dc:creator>
<dc:creator>Freimer, N. B.</dc:creator>
<dc:date>2017-07-07</dc:date>
<dc:identifier>doi:10.1101/160499</dc:identifier>
<dc:title><![CDATA[Whole Genome Sequencing in Psychiatric Disorders: the WGSPD Consortium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/162297v1?rss=1">
<title>
<![CDATA[
GPS2 regulates mitochondria biogenesis via mitochondrial retrograde signaling and chromatin remodeling of nuclear-encoded mitochondrial genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/162297v1?rss=1</link>
<description><![CDATA[
As most of the mitochondrial proteome is encoded in the nucleus, mitochondrial functions critically depend on nuclear gene expression and bidirectional mito-nuclear communication. However, mitochondria-to-nucleus communication pathways are incompletely understood. Here, we identify G-Protein Pathway Suppressor 2 (GPS2) as a mediator of mitochondrial retrograde signaling and a key transcriptional activator of nuclear-encoded mitochondrial genes in mammals. GPS2 regulated translocation from mitochondria to nucleus is essential for the transcriptional activation of the nuclear stress response to mitochondrial depolarization and for supporting basal mitochondrial biogenesis in differentiating adipocytes and in brown adipose tissue from mice. In the nucleus, GPS2 recruitment to target gene promoters regulates histone H3K9 demethylation and RNA Polymerase II (POL2) activation through inhibition of Ubc13-mediated ubiquitination. Together, these findings reveal an unexpected layer of regulation of mitochondrial gene transcription as they uncover a novel mitochondria-nuclear communication pathway.
]]></description>
<dc:creator>Cardamone, M. D.</dc:creator>
<dc:creator>Tanasa, B.</dc:creator>
<dc:creator>Cederquist, C.</dc:creator>
<dc:creator>Huang, J.</dc:creator>
<dc:creator>Mahdaviani, K.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Rosenfeld, M. G.</dc:creator>
<dc:creator>Liesa, M.</dc:creator>
<dc:creator>Perissi, V.</dc:creator>
<dc:date>2017-07-11</dc:date>
<dc:identifier>doi:10.1101/162297</dc:identifier>
<dc:title><![CDATA[GPS2 regulates mitochondria biogenesis via mitochondrial retrograde signaling and chromatin remodeling of nuclear-encoded mitochondrial genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/165316v1?rss=1">
<title>
<![CDATA[
Abnormal Speech Motor Control in Individuals with 16p11.2 Deletions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/165316v1?rss=1</link>
<description><![CDATA[
Speech and motor deficits are highly prevalent (>70%) in individuals with the 600 kb BP4-BP5 16p11.2 deletion; however, the mechanisms that drive these deficits are unclear, limiting our ability to target interventions and advance treatment. This study examined fundamental aspects of speech motor control in participants with the 16p11.2 deletion. To assess capacity for control of voice, we examined how accurately and quickly subjects changed the pitch of their voice within a trial to correct for a transient perturbation of the pitch of their auditory feedback. When compared to sibling controls, 16p11.2 deletion carriers show an over-exaggerated pitch compensation response to unpredictable mid-vocalization pitch perturbations. We also examined sensorimotor adaptation of speech by assessing how subjects learned to adapt their sustained productions of formants (speech spectral peak frequencies important for vowel identity), in response to consistent changes in their auditory feedback during vowel production. Deletion carriers show reduced sensorimotor adaptation to sustained vowel identity changes in auditory feedback. These results together suggest that 16p11.2 deletion carriers have fundamental impairments in the basic mechanisms of speech motor control and these impairments may partially explain the deficits in speech and language in these individuals.
]]></description>
<dc:creator>Demopoulos, C.</dc:creator>
<dc:creator>Kothare, H.</dc:creator>
<dc:creator>Mizuiri, D.</dc:creator>
<dc:creator>Henderson-Sabes, J.</dc:creator>
<dc:creator>Fregeau, B.</dc:creator>
<dc:creator>Tjernagel, J.</dc:creator>
<dc:creator>Houde, J.</dc:creator>
<dc:creator>Sherr, E.</dc:creator>
<dc:creator>Nagarajan, S. S.</dc:creator>
<dc:date>2017-07-18</dc:date>
<dc:identifier>doi:10.1101/165316</dc:identifier>
<dc:title><![CDATA[Abnormal Speech Motor Control in Individuals with 16p11.2 Deletions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/166124v1?rss=1">
<title>
<![CDATA[
An Automated Approach to the Quantitation of Vocalizations and Vocal Learning in the Songbird 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/166124v1?rss=1</link>
<description><![CDATA[
Studies of learning mechanisms critically depend on the ability to accurately assess learning outcomes. This assessment can be impeded by the often complex, multidimensional nature of behavior. We present a novel, automated approach to evaluating imitative learning that is founded in information theory. Conceptually, our approach estimates the amount of information present in a reference behavior that is absent from the learned behavior. We validate our approach through examination of songbird vocalizations, complex learned behaviors the study of which has provided many insights into sensory-motor learning in general and vocal learning in particular. Historically, learning has been holistically assessed by human inspection or through comparison of specific song features selected by experimenters (e.g. fundamental frequency, spectral entropy). In contrast, our approach relies on statistical models that broadly capture the structure of each song, and then uses these models to estimate the amount of information in the reference song but absent from the learned song. We show that our information theoretic measure of song learning (contrast entropy) is well correlated with human evaluation of song learning. We then expand the analysis beyond song learning and show that contrast entropy also detects the typical song deterioration that occurs following deafening. More broadly, this approach potentially provides a framework for assessing learning across a broad range of similarly structured behaviors.nnAuthor SummaryMeasuring learning outcomes is a critical objective of research into the neural, molecular, and behavioral mechanisms that support learning. Demonstration that a given manipulation results in better or worse learning outcomes requires an accurate and consistent measurement of learning quality. However, many behaviors (e.g. speech, walking, and reading) are complex and multidimensional, confounding the assessment of learning. One behavior subject to such confounds, vocal learning in Estrildid finches, has emerged as a vital model for sensory motor learning broadly and human speech learning in particular. Here, we demonstrate a new approach, founded in information theory, to the assessment of learning for complex high dimensional behaviors. Conceptually, we determine the amount of information (across many dimensions) present in a reference behavior and then assess how much of that information is present in the resultant learned behavior. We show that this measure provides an accurate, holistic, and automated assessment of vocal learning in Estrildid finches. Potentially, this same approach could be deployed to assess shared content in any multidimensional data, behavioral or otherwise.
]]></description>
<dc:creator>Mets, D. G.</dc:creator>
<dc:creator>Brainard, M. S.</dc:creator>
<dc:date>2017-07-20</dc:date>
<dc:identifier>doi:10.1101/166124</dc:identifier>
<dc:title><![CDATA[An Automated Approach to the Quantitation of Vocalizations and Vocal Learning in the Songbird]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/166983v1?rss=1">
<title>
<![CDATA[
BRG1/BRM-associated factor complex subunit diversity promotes temporally distinct gene expression programs in cardiogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/166983v1?rss=1</link>
<description><![CDATA[
Chromatin remodeling complexes instruct cellular differentiation and lineage specific transcription. The BRG1/BRM associated factor (BAF) complexes are important for several aspects of differentiation. We show that the catalytic subunit Brg1 has a specific role in cardiac precursors (CPs) to initiate cardiac gene expression programs and repress non-cardiac expression. Using immunoprecipitation with mass spectrometry (IP-MS), we determined the dynamic composition of BAF complexes during mammalian cardiac differentiation, and identified BAF60c (SMARCD3) and BAF170 (SMARCC2) as subunits enriched in CPs and cardiomyocytes (CM). Baf60c and Baf170 co-regulate gene expression with Brg1 in CPs, but in CMs control different gene expression programs, although still promoting a cardiac-specific gene set. BRG1, BAF60, and BAF170 all modulate chromatin accessibility, to either promote accessibility at activated genes, while closing up chromatin at repressed genes. BAF60c and BAF170 are required for proper BAF complex composition and stoichiometry, and promote BRG1 occupancy in CM. Additionally, BAF170 facilitates expulsion of BRG1-containing complexes in the transition from CP to CM. Thus, dynamic interdependent BAF complex subunit assembly modulates chromatin states and thereby directs temporal gene expression programs in cardiogenesis.nnSignificance statementBRG1/BRM associated factors (BAF) form multi-subunit protein complexes that reorganize chromatin and regulate transcription. Specific BAF complex subunits have important roles during cell differentiation and development. We systematically identify BAF subunit composition and find temporal enrichment of subunits during cardiomyocyte differentiation. We find the catalytic subunit BRG1 has important contributions in initiating gene expression programs in cardiac progenitors along with cardiac-enriched subunits BAF60c and BAF170. Both these proteins regulated BAF subunit composition and chromatin accessibility and prevent expression of non-cardiac developmental genes during precursor to cardiomyocyte differentiation. Mechanistically, we find BAF170 destabilizes the BRG1 complex and expels BRG1 from cardiomyocyte-specific genes. Thus, our data shows synergies between diverse BAF subunits in facilitating temporal gene expression programs during cardiogenesis.
]]></description>
<dc:creator>Hota, S.</dc:creator>
<dc:creator>Johnson, J.</dc:creator>
<dc:creator>Verschueren, E.</dc:creator>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Pennacchio, L.</dc:creator>
<dc:creator>Rossant, J.</dc:creator>
<dc:creator>Krogan, N.</dc:creator>
<dc:creator>Bruneau, B.</dc:creator>
<dc:date>2017-07-21</dc:date>
<dc:identifier>doi:10.1101/166983</dc:identifier>
<dc:title><![CDATA[BRG1/BRM-associated factor complex subunit diversity promotes temporally distinct gene expression programs in cardiogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/167239v1?rss=1">
<title>
<![CDATA[
The transcriptionally permissive chromatin state of ES cells is acutely tuned totranslational output 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/167239v1?rss=1</link>
<description><![CDATA[
A permissive chromatin environment coupled to hypertranscription is critical to drive the rapid proliferation of Embryonic Stem (ES) cells and peri-implantation embryos. We carried out a genome-wide screen to systematically dissect the regulation of the euchromatic state of ES cells. The results reveal that activity of cellular growth pathways, prominently protein synthesis, perpetuates the euchromatic state and hypertranscription of ES cells. Acute, mild inhibition of translation results in rapid depletion of euchromatic marks in ES cells and blastocysts, concurrent with delocalization of RNA polymerase II and reduction in nascent transcription. Remarkably, reduced translational output leads to rewiring of open chromatin within 3 hours, including decreased accessibility at a subset of active developmental enhancers and increased accessibility at histone genes and transposable elements. Using a proteome-scale analysis, we show that several euchromatin regulators are unstable proteins and thus continuously depend on a high translational output. We propose that this mechanistic interdependence of euchromatin, transcription and translation sets the pace of proliferation at peri-implantation and may be employed generally by stem/progenitor cells.
]]></description>
<dc:creator>Bulut-Karslioglu, A.</dc:creator>
<dc:creator>Macrae, T. A.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>Covarrubias, S.</dc:creator>
<dc:creator>Percharde, M.</dc:creator>
<dc:creator>Ku, G.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:creator>McManus, M. T.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Ramalho-santos, M.</dc:creator>
<dc:date>2017-07-22</dc:date>
<dc:identifier>doi:10.1101/167239</dc:identifier>
<dc:title><![CDATA[The transcriptionally permissive chromatin state of ES cells is acutely tuned totranslational output]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/167874v1?rss=1">
<title>
<![CDATA[
Tau Internalization is Regulated by 6-O Sulfation on Heparan Sulfate Proteoglycans (HSPGs) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/167874v1?rss=1</link>
<description><![CDATA[
The misfolding and accumulation of tau protein into intracellular aggregates known as neurofibrillary tangles is a pathological hallmark of neurodegenerative diseases such as Alzheimers disease. However, while tau propagation is a known marker for disease progression, exactly how tau propagates from one cell to another and what mechanisms govern this spread are still unclear. Here, we report that cellular internalization of tau is regulated by quaternary structure and have developed a cellular assay to screen for genetic modulators of tau uptake. Using CRISPRi technology we have tested 3200 genes for their ability to regulate tau entry and identified enzymes in the heparan sulfate proteoglycan biosynthetic pathway as key regulators. We show that 6-O-sulfation is critical for tauheparan sulfate interactions and that this modification regulates uptake in human central nervous system cell lines, iPS-derived neurons, and mouse organotypic brain slice culture. Together, these results suggest novel strategies to halt tau transmission.
]]></description>
<dc:creator>Rauch, J.</dc:creator>
<dc:creator>Chen, J. J.</dc:creator>
<dc:creator>Sorum, A. W.</dc:creator>
<dc:creator>Miller, G. M.</dc:creator>
<dc:creator>Sharf, T.</dc:creator>
<dc:creator>See, S. K.</dc:creator>
<dc:creator>Hsieh-Wilson, L. C.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Kosik, K. S.</dc:creator>
<dc:date>2017-07-24</dc:date>
<dc:identifier>doi:10.1101/167874</dc:identifier>
<dc:title><![CDATA[Tau Internalization is Regulated by 6-O Sulfation on Heparan Sulfate Proteoglycans (HSPGs)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/168187v1?rss=1">
<title>
<![CDATA[
A large kinome in a large cell: Stentor coeruleus possesses highly expanded kinase families and novel domain architectures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/168187v1?rss=1</link>
<description><![CDATA[
BackgroundStentor coeruleus is a large ciliated protist, about 1mm in length, with the extraordinary ability to fully regenerate each fragment after being cut into pieces, perfectly restoring cell polarity and morphology. Single-cell regeneration in Stentor remains one of the greatest long-standing mysteries of biology, but the recently published Stentor genome now enables studies on this organism at the molecular and genetic levels. Here we characterize the complete complement of kinases, or kinome, of Stentor, in order to begin to understand the signaling capacities that underlie Stentors unique biology.nnResultsThe genome of S. coeruleus contains over 2000 kinases, representing 6% of the predicted proteome. Classification of the kinase genes reveals large expansions in several kinase families, particularly in the CDPKs, the DYRKs, and in several mitotic kinase families including the PLKs, NEKs, and Auroras. The large expansion of the CDPK and DYRK kinase families is an unusual feature of the Stentor kinome compared to other ciliates with sequenced genomes. The DYRK family in Stentor, notably, contains only a single pseudokinase which may suggest an important role in Stentor growth and survival, while the smaller PEK family contains a novel pseudokinase subfamily. The Stentor kinome also has examples of new domain architectures that have not been previously observed in other organisms.nnConclusionOur analysis provides the first gene-level view into the signaling capabilities of Stentor and will lay the foundation for unraveling how this organism can coordinate processes as complex as regeneration throughout a giant cell.
]]></description>
<dc:creator>Reiff, S. B.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2017-07-25</dc:date>
<dc:identifier>doi:10.1101/168187</dc:identifier>
<dc:title><![CDATA[A large kinome in a large cell: Stentor coeruleus possesses highly expanded kinase families and novel domain architectures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/173815v1?rss=1">
<title>
<![CDATA[
Evidence for evolutionary shifts in the fitness landscape of human complex traits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/173815v1?rss=1</link>
<description><![CDATA[
Selection and mutation shape genetic variation underlying human traits, but the specific evolutionary mechanisms driving complex trait variation are largely unknown. We developed a statistical method that uses polarized GWAS summary statistics from a single population to detect signals of mutational bias and selection. We found evidence for non-neutral signals on variation underlying several traits (BMI, schizophrenia, Crohns disease, educational attainment, and height). We then used simulations that incorporate simultaneous negative and positive selection to show that these signals are consistent with mutational bias and shifts in the fitness-phenotype relationship, but not stabilizing selection or mutational bias alone. We additionally replicate two of our top three signals (BMI and educational attainment) in an external cohort, and show that population stratification may have confounded GWAS summary statistics for height in the GIANT cohort. Our results provide a flexible and powerful framework for evolutionary analysis of complex phenotypes in humans and other species, and offer insights into the evolutionary mechanisms driving variation in human polygenic traits.nnImpact summaryMany traits are variable within human populations and are likely to have a substantial and complex genetic component. This implies that mutations that have a functional impact on complex human traits have arisen throughout our species evolutionary history. However, it remains unclear how processes such as natural selection may have acted to shape trait variation at the genetic and phenotypic level. Better understanding of the mechanisms driving trait variation could provide insights into our evolutionary past and help clarify why it has been so difficult to map the preponderance of causal variation for common heritable diseases.nnIn this study, we developed and applied methods for detecting signatures of mutation bias (i.e., the propensity of a new variant to be either trait-increasing or trait-decreasing) and natural selection acting on trait variation. We applied our approach to several heritable traits, and found evidence for both natural selection and mutation bias, including selection for decreased BMI and decreased risk for Crohns disease and schizophrenia.nnWhile our results are consistent with plausible evolutionary scenarios shaping a range of traits, it should be noted that the field of polygenic selection detection is still new, and current methods (including ours) rely on data from genome-wide association studies (GWAS). The data produced by these studies may be vulnerable to certain cryptic biases, especially population stratification, which could induce false selection signals. We therefore repeated our analyses for the top three hits in a cohort that should be less susceptible to this problem - we found that two of our top three signals replicated (BMI and educational attainment), while height did not. Our results highlight both the promise and pitfalls of polygenic selection detection approaches, and suggest a need for further work disentangling stratification from selection.
]]></description>
<dc:creator>Uricchio, L. H.</dc:creator>
<dc:creator>Kitano, H. C.</dc:creator>
<dc:creator>Gusev, A.</dc:creator>
<dc:creator>Zaitlen, N. A.</dc:creator>
<dc:date>2017-08-08</dc:date>
<dc:identifier>doi:10.1101/173815</dc:identifier>
<dc:title><![CDATA[Evidence for evolutionary shifts in the fitness landscape of human complex traits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/179242v1?rss=1">
<title>
<![CDATA[
Progressive reconfiguration of resting-state brain networks as psychosis develops: Preliminary results from the North American Prodrome Longitudinal Study (NAPLS) consortium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/179242v1?rss=1</link>
<description><![CDATA[
Mounting evidence has shown disrupted brain network architecture across the psychosis spectrum. However, whether these changes relate to the development of psychosis is unclear. Here, we used graph theoretical analysis to investigate longitudinal changes in resting-state brain networks in samples of 72 subjects at clinical high risk (including 8 cases who converted to full psychosis) and 48 healthy controls drawn from the North American Prodrome Longitudinal Study (NAPLS) consortium. We observed progressive reduction in global efficiency (P = 0.006) and increase in network diversity (P = 0.001) in converters compared with non-converters and controls. More refined analysis separating nodes into nine key brain networks demonstrated that these alterations were primarily driven by progressively diminished local efficiency in the default-mode network (P = 0.004) and progressively enhanced node diversity across all networks (P < 0.05). The change rates of network efficiency and network diversity were significantly correlated (P = 0.003), suggesting these changes may reflect shared underlying neural mechanisms. In addition, change rates of global efficiency and node diversity were significantly correlated with change rate of cortical thinning in the prefrontal cortex in converters (P < 0.03) and could be predicted by visuospatial memory scores at baseline (P < 0.04). These results provide preliminary evidence for longitudinal reconfiguration of resting-state brain networks during psychosis development and suggest that decreased network efficiency, reflecting an increase in path length between nodes, and increased network diversity, reflecting a decrease in the consistency of functional network organization, are implicated in the progression to full psychosis.
]]></description>
<dc:creator>Cao, H.</dc:creator>
<dc:creator>Chung, Y.</dc:creator>
<dc:creator>McEwen, S.</dc:creator>
<dc:creator>Bearden, C.</dc:creator>
<dc:creator>Addington, J.</dc:creator>
<dc:creator>Goodyear, B.</dc:creator>
<dc:creator>Cadenhead, K.</dc:creator>
<dc:creator>Mirzakhanian, H.</dc:creator>
<dc:creator>Cornblatt, B.</dc:creator>
<dc:creator>Olvet, D.</dc:creator>
<dc:creator>Mathalon, D.</dc:creator>
<dc:creator>McGlashan, T.</dc:creator>
<dc:creator>Perkins, D.</dc:creator>
<dc:creator>Belger, A.</dc:creator>
<dc:creator>Seidman, L.</dc:creator>
<dc:creator>Thermenos, H.</dc:creator>
<dc:creator>Tsuang, M.</dc:creator>
<dc:creator>van Erp, T.</dc:creator>
<dc:creator>Walker, E.</dc:creator>
<dc:creator>Hamann, S.</dc:creator>
<dc:creator>Anticevic, A.</dc:creator>
<dc:creator>Woods, S.</dc:creator>
<dc:creator>Cannon, T.</dc:creator>
<dc:date>2017-08-21</dc:date>
<dc:identifier>doi:10.1101/179242</dc:identifier>
<dc:title><![CDATA[Progressive reconfiguration of resting-state brain networks as psychosis develops: Preliminary results from the North American Prodrome Longitudinal Study (NAPLS) consortium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/180620v1?rss=1">
<title>
<![CDATA[
BBSome trains remove activated GPCRs from cilia by enabling passage through the transition zone 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/180620v1?rss=1</link>
<description><![CDATA[
A diffusion barrier at the transition zone enables the compartmentalization of signaling molecules by cilia. The BBSome and the small GTPase Arl6, which triggers BBSome coat polymerization, are required for the exit of activated signaling receptors from cilia, but how diffusion barriers are crossed when membrane proteins exit cilia remains to be determined. Here we found that activation of the ciliary GPCRs Smoothened and SSTR3 drove the Arl6-dependent assembly of large, highly processive and cargo-laden retrograde BBSome trains. Single-molecule imaging revealed that the assembly of BBSome trains enables the lateral transport of ciliary GPCRs across the transition zone. Yet, the removal of activated GPCRs from cilia was inefficient because a second, periciliary diffusion barrier was infrequently crossed. We conclude that exit from cilia is a two-step process in which BBSome/Arl6 trains first moves activated GPCRs through the transition zone before a periciliary barrier can be crossed.nnSummaryUpon activation, GPCRs must exit cilia for appropriate signal transduction. Using bulk imaging of BBSome and single molecule imaging of GPCRs, Ye et al. demonstrate that retrograde BBSome trains assemble on-demand upon GPCR activation and ferry GPCRs across the transition zone. Yet, ciliary exit often fails because of a second diffusion barrier.
]]></description>
<dc:creator>Ye, F.</dc:creator>
<dc:creator>Nager, A. R.</dc:creator>
<dc:creator>Nachury, M. V.</dc:creator>
<dc:date>2017-08-26</dc:date>
<dc:identifier>doi:10.1101/180620</dc:identifier>
<dc:title><![CDATA[BBSome trains remove activated GPCRs from cilia by enabling passage through the transition zone]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/181248v1?rss=1">
<title>
<![CDATA[
Genomic and proteomic analysis of Human herpesvirus 6 reveals distinct clustering of acute versus inherited forms and reannotation of reference strain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/181248v1?rss=1</link>
<description><![CDATA[
Human herpesvirus-6A and -6B (HHV-6) are betaherpesviruses that reach >90% seroprevalence in the adult population. Unique among human herpesviruses, HHV-6 can integrate into the subtelomeric regions of human chromosomes; when this occurs in germ line cells it causes a condition called inherited chromosomally integrated HHV-6 (iciHHV-6). To date, only two complete genomes are available for HHV-6B. Using a custom capture panel for HHV-6B, we report near-complete genomes from 61 isolates of HHV-6B from active infections (20 from Japan, 35 from New York state, and 6 from Uganda), and 64 strains of iciHHV-6B (mostly from North America). We also report partial genome sequences from 10 strains of iciHHV-6A. Although the overall sequence diversity of HHV-6 is limited relative to other human herpesviruses, our sequencing identified geographical clustering of HHV-6B sequences from active infections, as well as evidence of recombination among HHV-6B strains. One strain of active HHV-6B was more divergent than any other HHV-6B previously sequenced. In contrast to the active infections, sequences from iciHHV-6 cases showed reduced sequence diversity. Strikingly, multiple iciHHV-6B sequences from unrelated individuals were found to be completely identical, consistent with a founder effect. However, several iciHHV-6B strains intermingled with strains from active pediatric infection, consistent with the hypothesis that intermittent de novo integration into host germline cells can occur during active infection Comparative genomic analysis of the newly sequenced strains revealed numerous instances where conflicting annotations between the two existing reference genomes could be resolved. Combining these findings with transcriptome sequencing and shotgun proteomics, we reannotated the HHV-6B genome and found multiple instances of novel splicing and genes that hitherto had gone unannotated. The results presented here constitute a significant genomic resource for future studies on the detection, diversity, and control of HHV-6.nnAuthor SummaryHHV-6 is a ubiquitous large DNA virus that is the most common cause of febrile seizures and reactivates in allogeneic stem cell patients. It also has the unique ability among human herpesviruses to be integrated into the genome of every cell via integration in the germ line, a condition called inherited chromosomally integrated (ici)HHV-6, which affects approximately 1% of the population. To date, very little is known about the comparative genomics of HHV-6. We sequenced 61 isolates of HHV-6B from active infections, 64 strains of iciHHV-6B, and 10 strains of iciHHV-6A. We found geographic clustering of HHV-6B strains from active infections. In contrast, iciHHV-6B had reduced sequence diversity, with many identical sequences of iciHHV-6 found in individuals not known to share recent common ancestry, consistent with a founder effect from a remote common ancestor with iciHHV-6. We also combined our genomic analysis with transcriptome sequencing and shotgun proteomics to correct previous misannotations of the HHV-6 genome.
]]></description>
<dc:creator>Greninger, A.</dc:creator>
<dc:creator>Knudsen, G.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Hanson, D.</dc:creator>
<dc:creator>Hall Sedlak, R. A.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Guan, J.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Peddu, V.</dc:creator>
<dc:creator>Boeckh, M.</dc:creator>
<dc:creator>Huang, M.-L.</dc:creator>
<dc:creator>Cook, L.</dc:creator>
<dc:creator>Depledge, D.</dc:creator>
<dc:creator>Zerr, D.</dc:creator>
<dc:creator>Koelle, D.</dc:creator>
<dc:creator>Gantt, S.</dc:creator>
<dc:creator>Yoshikawa, T.</dc:creator>
<dc:creator>Caserta, M.</dc:creator>
<dc:creator>Hill, J.</dc:creator>
<dc:creator>Jerome, K.</dc:creator>
<dc:date>2017-08-27</dc:date>
<dc:identifier>doi:10.1101/181248</dc:identifier>
<dc:title><![CDATA[Genomic and proteomic analysis of Human herpesvirus 6 reveals distinct clustering of acute versus inherited forms and reannotation of reference strain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/181859v1?rss=1">
<title>
<![CDATA[
Human demographic history has amplified the effects background selection across the genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/181859v1?rss=1</link>
<description><![CDATA[
Natural populations often grow, shrink, and migrate over time. Demographic processes such as these can impact genome-wide levels of genetic diversity. In addition, genetic variation in functional regions of the genome can be altered by natural selection, which drives adaptive mutations to higher frequencies or purges deleterious ones. Such selective processes impact not only the sites directly under selection but also nearby neutral variation through genetic linkage through processes referred to as genetic hitch-hiking in the context of positive selection and background selection (BGS) in the context of purifying selection. While there is extensive literature examining the impact of selection at linked sites at demographic equilibrium, less is known about how non-equilibrium demographic processes impact the effects of hitchhiking and BGS. Utilizing a global sample of human whole-genome sequences from the Thousand Genomes Project and extensive simulations, we investigate how non-equilibrium demographic processes magnify and dampen the consequences of selection at linked sites across the human genome. When binning the genome by inferred strength of BGS, we observe that, compared to Africans, non-African populations have experienced larger proportional decreases in neutral genetic diversity in such regions. We replicate these findings in admixed populations by showing that non-African ancestral components of the genome have also been impacted more severely in these regions. We attribute these differences to the strong, sustained/recurrent population bottlenecks that non-Africans experienced as they migrated out of Africa and throughout the globe. Furthermore, we observe a strong correlation between FST and inferred strength of BGS, suggesting a stronger rate of genetic drift. Forward simulations of human demographic history with a model of BGS support these observations. Our results show that non-equilibrium demography significantly alters the consequences selection at linked sites and support the need for more work investigating the dynamic process of multiple evolutionary forces operating in concert.nnAuthor summaryPatterns of genetic diversity within a species are affected at broad and fine scales by population size changes ("demography") and natural selection. From both population genetics theory and observation of genomic sequence data, it is known that demography can alter genome-wide average neutral genetic diversity. Additionally, natural selection can affect neutral genetic diversity regionally across the genome via selection at linked sites. During this process, natural selection acting on adaptive or deleterious variants in the genome will also impact diversity at nearby neutral sites due to genetic linkage. However, less is well known about the dynamic changes to diversity that occur in regions impacted by selection at linked sites when a population undergoes a size change. We characterize these dynamic changes using thousands of human genomes and find that the population size changes experienced by humans have shaped the consequences of linked selection across the genome. In particular, population contractions, such as those experienced by non-Africans, have disproportionately decreased neutral diversity in regions of the genome inferred to be under strong background selection (i.e., selection at linked sites that is caused by natural selection acting on deleterious variants), resulting in large differences between African and non-African populations.
]]></description>
<dc:creator>Torres, R.</dc:creator>
<dc:creator>Szpiech, Z. A.</dc:creator>
<dc:creator>Hernandez, R. D.</dc:creator>
<dc:date>2017-08-29</dc:date>
<dc:identifier>doi:10.1101/181859</dc:identifier>
<dc:title><![CDATA[Human demographic history has amplified the effects background selection across the genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/183418v1?rss=1">
<title>
<![CDATA[
Correction of autoimmune IL2RA mutations in primary human T cells using non-viral genome targeting 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/183418v1?rss=1</link>
<description><![CDATA[
Human T cells are central to physiological immune homeostasis, which protects us from pathogens without collateral autoimmune inflammation. They are also the main effectors in most current cancer immunotherapy strategies1. Several decades of work have aimed to genetically reprogram T cells for therapeutic purposes2-5, but as human T cells are resistant to most standard methods of large DNA insertion these approaches have relied on recombinant viral vectors, which do not target transgenes to specific genomic sites6, 7. In addition, the need for viral vectors has slowed down research and clinical use as their manufacturing and testing is lengthy and expensive. Genome editing brought the promise of specific and efficient insertion of large transgenes into target cells through homology-directed repair (HDR), but to date in human T cells this still requires viral transduction8, 9. Here, we developed a non-viral, CRISPR-Cas9 genome targeting system that permits the rapid and efficient insertion of individual or multiplexed large (>1 kilobase) DNA sequences at specific sites in the genomes of primary human T cells while preserving cell viability and function. We successfully tested the potential therapeutic use of this approach in two settings. First, we corrected a pathogenic IL2RA mutation in primary T cells from multiple family members with monogenic autoimmune disease and demonstrated enhanced signalling function. Second, we replaced the endogenous T cell receptor (TCR) locus with a new TCR redirecting T cells to a cancer antigen. The resulting TCR-engineered T cells specifically recognized the tumour antigen, with concomitant cytokine release and tumour cell killing. Taken together, these studies provide preclinical evidence that non-viral genome targeting will enable rapid and flexible experimental manipulation and therapeutic engineering of primary human immune cells.
]]></description>
<dc:creator>Roth, T. L.</dc:creator>
<dc:creator>Yu, R.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Hiatt, J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Schumann, K.</dc:creator>
<dc:creator>Tobin, V.</dc:creator>
<dc:creator>Ferris, A. M.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Schickel, J.-N.</dc:creator>
<dc:creator>Pellerin, L.</dc:creator>
<dc:creator>Carmody, D.</dc:creator>
<dc:creator>Alkorta-Aranburu, G.</dc:creator>
<dc:creator>Del Gaudio, D.</dc:creator>
<dc:creator>Cho, M.</dc:creator>
<dc:creator>Matsumoto, H.</dc:creator>
<dc:creator>Morell, M.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Nguyen, D.</dc:creator>
<dc:creator>Quadros, R.</dc:creator>
<dc:creator>Gurumurthy, C.</dc:creator>
<dc:creator>Smith, B.</dc:creator>
<dc:creator>Haugwitz, M.</dc:creator>
<dc:creator>Hughes, S. H.</dc:creator>
<dc:creator>Weissman, J.</dc:creator>
<dc:creator>May, A. P.</dc:creator>
<dc:creator>Kupfer, G.</dc:creator>
<dc:creator>Greeley, S.</dc:creator>
<dc:creator>Bacchetta, R.</dc:creator>
<dc:creator>Meffre, E.</dc:creator>
<dc:creator>Roncarolo, M. G.</dc:creator>
<dc:creator>Romberg, N.</dc:creator>
<dc:creator>Herold, K. C.</dc:creator>
<dc:creator>Leonetti, M. D.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2017-08-31</dc:date>
<dc:identifier>doi:10.1101/183418</dc:identifier>
<dc:title><![CDATA[Correction of autoimmune IL2RA mutations in primary human T cells using non-viral genome targeting]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/185009v1?rss=1">
<title>
<![CDATA[
Rescue of Conformational Dynamics in Enzyme Catalysis by Directed Evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/185009v1?rss=1</link>
<description><![CDATA[
Rational design and directed evolution have proved to be successful approaches to increase catalytic efficiencies of both natural and artificial enzymes1-3. However, a comprehensive understanding of how evolution shapes the energy landscape of catalysis remains a fundamental challenge. Protein dynamics is widely recognized as important, but due to the inherent flexibility of biological macromolecules it is often difficult to distinguish which conformational changes are directly related to function. Here, we used directed evolution on an impaired mutant of the human proline isomerase cyclophilin A (CypA) and identify two second-shell mutations that partially restore its catalytic activity. We show both kinetically, using NMR spectroscopy, and structurally, by room-temperature X-ray crystallography, how local perturbations propagate through a large allosteric network to facilitate conformational dynamics. The increased catalysis selected for in the evolutionary screen could be rationalized entirely by accelerated interconversion between the two catalytically essential conformational sub-states, which are both captured in the high-resolution X-ray ensembles at room temperature. Our data provide a glimpse of the evolutionary trajectory of an enzymes energy landscape and shows how subtle changes can fine-tune its function.
]]></description>
<dc:creator>Otten, R.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Kenner, L. R.</dc:creator>
<dc:creator>Clarkson, M. W.</dc:creator>
<dc:creator>Mavor, D.</dc:creator>
<dc:creator>Tawfik, D. S.</dc:creator>
<dc:creator>Kern, D.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2017-09-05</dc:date>
<dc:identifier>doi:10.1101/185009</dc:identifier>
<dc:title><![CDATA[Rescue of Conformational Dynamics in Enzyme Catalysis by Directed Evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/189795v1?rss=1">
<title>
<![CDATA[
Phylogeny-corrected identification of microbial gene families relevant to human gut colonization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/189795v1?rss=1</link>
<description><![CDATA[
The mechanisms by which different microbes colonize the healthy human gut versus other body sites, the gut in disease states, or other environments remain largely unknown. Identifying microbial genes influencing fitness in the gut could lead to new ways to engineer probiotics or disrupt pathogenesis. We approach this problem by measuring the statistical association between having a species having a gene and the probability that the species is present in the gut microbiome. The challenge is that closely related species tend to be jointly present or absent in the microbiome and also share many genes, only a subset of which are involved in gut adaptation. We show that this phylogenetic correlation indeed leads to many false discoveries and propose phylogenetic linear regression as a powerful solution. To apply this method across the bacterial tree of life, where most species have not been experimentally phenotyped, we use metagenomes from hundreds of people to quantify each species prevalence in and specificity for the gut microbiome. This analysis reveals thousands of genes potentially involved in adaptation to the gut across species, including many novel candidates as well as processes known to contribute to fitness of gut bacteria, such as acid tolerance in Bacteroidetes and sporulation in Firmicutes. We also find microbial genes associated with a preference for the gut over other body sites, which are significantly enriched for genes linked to fitness in an in vivo competition experiment. Finally, we identify gene families associated with higher prevalence in patients with Crohns disease, including Proteobacterial genes involved in conjugation and fimbria regulation, processes previously linked to inflammation. These gene targets may represent new avenues for modulating host colonization and disease. Our strategy of combining metagenomics with phylogenetic modeling is general and can be used to identify genes associated with adaptation to any environment.nnAuthor SummaryWhy do certain microbes and not others colonize our gut, and why do they differ between healthy and sick people? One explanation is the genes in their genomes. If we can find microbial genes involved in gut adaptation, we may be able to keep out pathogens and encourage the growth of beneficial microbes. One could look for genes that were present more often in prevalent microbes, and less often in rare ones.nnHowever, this ignores that related species are more likely to share an environment and also share many unrelated phenotypes simply because of common ancestry. To solve this problem, we used a method from ecology that accounts for phylogenetic relatedness. We first calculated gut prevalence for thousands of species using a compendium of shotgun sequencing data, then tested for genes associated with prevalence, adjusting for phylogenetic relationships. We found genes that are associated with overall gut prevalence, with a preference for the gut over other body sites, and with the gut in Crohns disease versus health. Many of these findings have biological plausibility based on existing literature. We also showed agreement with the results of a previously published high-throughput screen of bacterial gene knockouts in mice. These results, and this type of analysis, may eventually lead to new strategies for maintaining gut health.
]]></description>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:creator>Nayfach, S.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2017-09-16</dc:date>
<dc:identifier>doi:10.1101/189795</dc:identifier>
<dc:title><![CDATA[Phylogeny-corrected identification of microbial gene families relevant to human gut colonization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/200212v1?rss=1">
<title>
<![CDATA[
The geometry of the distance-decay of similarity in ecological communities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/200212v1?rss=1</link>
<description><![CDATA[
Understanding beta-diversity has strong implications for evaluating the extent of biodiversity and formulating effective conservation policy. Here, we show that the distance-decay relationship, an important measure of beta-diversity, follows a universal form which we call the piecewise quadratic model. To derive the piecewise quadratic model, we develop a new conceptual framework which is based on geometric probability and several key insights about the roles of study design (e.g., plot dimensions and spatial distributions). We fit the piecewise quadratic model to six empirical distance-decay relationships, spanning a range of taxa and spatial scales, including surveys of tropical vegetation, mammals, and amphibians. We find that the model predicts the functional form of the relationships extremely well, with coefficients of determination in excess of 0.95. Moreover, the model predicts a phase transition at distance scales where sample plots are overlapping, which we confirm empirically. Our framework and model provide a fundamental, quantitative link between distance-decay relationships and the shapes of ranges of taxa.
]]></description>
<dc:creator>Ladau, J.</dc:creator>
<dc:creator>Green, J. L.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2017-10-09</dc:date>
<dc:identifier>doi:10.1101/200212</dc:identifier>
<dc:title><![CDATA[The geometry of the distance-decay of similarity in ecological communities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.08.868828v1?rss=1">
<title>
<![CDATA[
Heterogeneity and targeted therapy-induced adaptations in lung cancer revealed by longitudinal single-cell RNA sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.08.868828v1?rss=1</link>
<description><![CDATA[
Lung cancer, the leading cause of cancer mortality, exhibits heterogeneity that enables adaptability, limits therapeutic success, and remains incompletely understood. Single-cell RNA sequencing (scRNAseq) of metastatic lung cancer was performed using 44 tumor biopsies obtained longitudinally from 27 patients before and during targeted therapy. Over 20,000 cancer and tumor microenvironment (TME) single-cell profiles exposed a rich and dynamic tumor ecosystem. scRNAseq of cancer cells illuminated targetable oncogenes beyond those detected clinically. Cancer cells surviving therapy as residual disease (RD) expressed an alveolar-regenerative cell signature suggesting a therapy-induced primitive cell state transition, whereas those present at on-therapy progressive disease (PD) upregulated kynurenine, plasminogen, and gap junction pathways. Active T-lymphocytes and decreased macrophages were present at RD and immunosuppressive cell states characterized PD. Biological features revealed by scRNAseq were biomarkers of clinical outcomes in independent cohorts. This study highlights how therapy-induced adaptation of the multi-cellular ecosystem of metastatic cancer shapes clinical outcomes.
]]></description>
<dc:creator>Maynard, A.</dc:creator>
<dc:creator>McCoach, C. E.</dc:creator>
<dc:creator>Rotow, J. K.</dc:creator>
<dc:creator>Harris, L.</dc:creator>
<dc:creator>Haderk, F.</dc:creator>
<dc:creator>Kerr, L. D.</dc:creator>
<dc:creator>Yu, E. A.</dc:creator>
<dc:creator>Schenk, E. L.</dc:creator>
<dc:creator>Tan, W.</dc:creator>
<dc:creator>Zee, A.</dc:creator>
<dc:creator>Tan, M.</dc:creator>
<dc:creator>Gui, P.</dc:creator>
<dc:creator>Lea, T.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Urisman, A.</dc:creator>
<dc:creator>Jones, K.</dc:creator>
<dc:creator>Sit, R.</dc:creator>
<dc:creator>Kolli, P. K.</dc:creator>
<dc:creator>Seeley, E.</dc:creator>
<dc:creator>Gesthalter, Y.</dc:creator>
<dc:creator>Le, D. D.</dc:creator>
<dc:creator>Yamauchi, K. A.</dc:creator>
<dc:creator>Naeger, D. M.</dc:creator>
<dc:creator>Thomas, N. J.</dc:creator>
<dc:creator>Gupta, A.</dc:creator>
<dc:creator>Gonzalez, M.</dc:creator>
<dc:creator>Do, H.</dc:creator>
<dc:creator>Tan, L.</dc:creator>
<dc:creator>Gomez-Sjoberg, R.</dc:creator>
<dc:creator>Gubens, M.</dc:creator>
<dc:creator>Jahan, T.</dc:creator>
<dc:creator>Kratz, J. R.</dc:creator>
<dc:creator>Jablons, D.</dc:creator>
<dc:creator>Neff, N.</dc:creator>
<dc:creator>Doebele, R. C.</dc:creator>
<dc:creator>Weissman, J.</dc:creator>
<dc:creator>Blakely, C. M.</dc:creator>
<dc:creator>Darmanis, S.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:date>2019-12-13</dc:date>
<dc:identifier>doi:10.1101/2019.12.08.868828</dc:identifier>
<dc:title><![CDATA[Heterogeneity and targeted therapy-induced adaptations in lung cancer revealed by longitudinal single-cell RNA sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.13.875112v1?rss=1">
<title>
<![CDATA[
Systems analysis of blood transcriptomes in dementia patients reveals an innate immune response shared across disorders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.13.875112v1?rss=1</link>
<description><![CDATA[
The role of peripheral inflammation in dementia is an important but complex topic. We present here the largest cohort of peripheral blood gene expression data ever assembled from patients with dementia and matching controls. Importantly, this cohort includes individuals from a diverse set of dementia disorders, including Alzheimers Disease (AD), mild cognitive impairment (MCI), and multiple disorders within the frontotemporal dementia (FTD) spectrum. We found strong transcriptional evidence of an innate immune inflammatory response, mediated by monocytes and neutrophils, in AD, MCI, and two FTD subtypes, PSP and nfvPPA. This transcriptional inflammatory response is enriched for genetic risk for AD, in part because it is also enriched for microglial genes, which have previously been implicated in AD risk. Finally, we show that this transcriptional response is strongly enriched for binding of the transcription factors PU.1 and RELA, which have previously been linked to AD risk and progression.
]]></description>
<dc:creator>Nachun, D.</dc:creator>
<dc:creator>Ramos, E. M.</dc:creator>
<dc:creator>Karydas, A.</dc:creator>
<dc:creator>Dokuru, D.</dc:creator>
<dc:creator>Gao, F.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Van Berlo, V.</dc:creator>
<dc:creator>Sears, R. L.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Boxer, A. L.</dc:creator>
<dc:creator>Rosen, H. H.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Coppola, G.</dc:creator>
<dc:date>2019-12-13</dc:date>
<dc:identifier>doi:10.1101/2019.12.13.875112</dc:identifier>
<dc:title><![CDATA[Systems analysis of blood transcriptomes in dementia patients reveals an innate immune response shared across disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.18.881680v1?rss=1">
<title>
<![CDATA[
Differential Correlation of Serum BDNF and microRNA Content in Rats with Rapid or Late Onset of Heavy Alcohol Use 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.18.881680v1?rss=1</link>
<description><![CDATA[
Heavy alcohol use reduces the levels of the brain-derived neurotrophic factor (BDNF) in the prefrontal cortex of rodents through the upregulation of microRNAs targeting BDNF mRNA. In humans, an inverse correlation exists between circulating blood levels of BDNF and the severity of psychiatric disorders including alcohol abuse. Here, we set out to determine whether a history of heavy alcohol use produces comparable alterations in the blood of rats. We used an intermittent access to 20% alcohol using the 2-bottle choice paradigm (IA20%2BC), and measured circulating levels of BDNF protein and microRNAs in the serum of Long-Evans rats before and after 8-weeks of excessive alcohol intake. We observed that the drinking profile of heavy alcohol users is not unified; Whereas 70% of the rats gradually escalate their alcohol intake (Late Onset), 30% of alcohol users exhibit a very Rapid Onset of drinking (Rapid Onset). We found that serum BDNF levels are negatively correlated with alcohol intake in both Rapid Onset and Late Onset rats. In contrast, increased expression of the microRNAs (miRs) targeting BDNF, miR30a-5p, miR-195-5p, miR191-5p and miR206-3p, was detected only in the Rapid Onset rats. Finally, we report that the alcohol-dependent molecular changes are not due to alterations in platelet number. Our data suggest that rats exhibit both Late and Rapid Onset of alcohol intake. We further show that heavy alcohol use produces comparable changes in BDNF protein levels in both groups. However, circulating microRNAs are responsive to alcohol only in the Rapid Onset rats.
]]></description>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>phamluong, K.</dc:creator>
<dc:creator>Darevesky, D.</dc:creator>
<dc:creator>Welman, M.</dc:creator>
<dc:creator>Moffat, J.</dc:creator>
<dc:creator>Sakhai, S.</dc:creator>
<dc:creator>Whiteley, E.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:creator>Laguesse, S.</dc:creator>
<dc:creator>Farokhnia, M.</dc:creator>
<dc:creator>Leggio, L.</dc:creator>
<dc:creator>Lordkipanidze, M.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2019-12-19</dc:date>
<dc:identifier>doi:10.1101/2019.12.18.881680</dc:identifier>
<dc:title><![CDATA[Differential Correlation of Serum BDNF and microRNA Content in Rats with Rapid or Late Onset of Heavy Alcohol Use]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.23.878207v1?rss=1">
<title>
<![CDATA[
Lactoferrin Reverses Methotrexate Driven Epithelial Barrier Defect by Inhibiting TGF-β Mediated Epithelial to Mesenchymal Transition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.23.878207v1?rss=1</link>
<description><![CDATA[
BACKGROUND AND AIMSMethotrexate is an important tool in the arsenal of oncologists, gastroenterologists, and rheumatologists. At low doses it induces intestinal barrier dysfunction that may induce side effects such as gastrointestinal discomfort and liver injury. Previous studies suggest that lactoferrin can improve barrier function in a variety of contexts. This study set out to determine the mechanism of methotrexate induced barrier dysfunction and assess the effect of lactoferrin and other components of human breast milk on this dysfunction.

METHODSUsing a murine enteroid model and Caco2 spheroids, we measured flux of basolateral-administered fluorescent dextran into the lumen. Barrier dysfunction was induced using methotrexate (220 nM) or lipopolysaccharide (20 nM). Human lactoferrin was added at 0.8 mg/ml (10 {micro}M). RNAseq was performed on exposed samples.

RESULTSLactoferrin blocks methotrexate-induced barrier dysfunction in murine enteroids. Similar results were observed when barrier dysfunction was induced in Caco2 spheroids with methotrexate and LPS, but not ML7. RNAseq revealed activation of TGF-{beta} response genes and epithelial-mesenchymal transition (EMT) by methotrexate, which normalized in the presence of lactoferrin. TGF-{beta} receptor inhibition (RepSox) blocked methotrexate induced barrier dysfunction in Caco2 spheroids. 20 nM TGF-{beta} induced barrier dysfunction in Caco2 spheroids which was also inhibited by lactoferrin.

CONCLUSIONSMethotrexate induces barrier dysfunction by activation of an EMT program promoted by TGF-{beta} signaling and inhibited by lactoferrin. Lactoferrin is also protective of barrier function in an LPS-induced model. The likely mechanism of this effect is blockade of EMT programs induced by TGF-{beta}.
]]></description>
<dc:creator>Wallach, T.</dc:creator>
<dc:creator>Srivastava, V.</dc:creator>
<dc:creator>Reyes, E.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:date>2019-12-26</dc:date>
<dc:identifier>doi:10.1101/2019.12.23.878207</dc:identifier>
<dc:title><![CDATA[Lactoferrin Reverses Methotrexate Driven Epithelial Barrier Defect by Inhibiting TGF-β Mediated Epithelial to Mesenchymal Transition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.25.888586v1?rss=1">
<title>
<![CDATA[
LegoBody: facile generation of bispecific and multi-specific antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.25.888586v1?rss=1</link>
<description><![CDATA[
Bispecific and multi-specific antibodies are capable of recognizing multiple ligands simultaneously or synergistically, creating complex biological interactions not achievable by monoclonal antibodies, thus expanding opportunities for novel therapy development. With the large number of monoclonal antibodies either approved or under clinical development, there are numerous opportunities to combine their specificities to further improve therapeutic potential. Although simple in concept, clinical development of bi- and multi-specific antibodies face several challenges, chief of which is how to efficiently and reliably produce bispecific and multi-specific antibodies with expected specificity and desired biophysical properties. In this study, we developed a modular approach that uses temporary linkers to enforce proper chain pairing and proteases such as thrombin to remove those linkers from the final product. Combined with the  knob-into-hole design, we can generate IgG-like, bi- or multi-specific antibodies from any pre-existing monoclonal antibodies. The approach is highly versatile and applicable to any monoclonal antibody pair or panel, expediting evaluation and therapeutic development of bi- and multi-specific antibodies.
]]></description>
<dc:creator>Lang, S.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Bidlingmaier, S.</dc:creator>
<dc:creator>Lee, N.-K.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:date>2019-12-27</dc:date>
<dc:identifier>doi:10.1101/2019.12.25.888586</dc:identifier>
<dc:title><![CDATA[LegoBody: facile generation of bispecific and multi-specific antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.26.888933v1?rss=1">
<title>
<![CDATA[
Selective and invariant features of neural response surfaces measured with principal curvature 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.26.888933v1?rss=1</link>
<description><![CDATA[
The responses of most visual cortical neurons are highly nonlinear functions of image stimuli. With the sparse coding network, a recurrent model of V1 computation, we apply techniques from differential geometry to these nonlinear responses and classify them as forms of selectivity or invariance. The selectivity and invariance of responses of individual neurons are quantified by measuring the principal curvatures of neural response surfaces in high-dimensional image space. An extended two-layer version of the network model that captures some properties of higher visual cortical areas is also characterized using this approach. We argue that this geometric view allows for the quantification of feature selectivity and invariance in network models in a way that provides insight into the computations necessary for object recognition.
]]></description>
<dc:creator>Golden, J. R.</dc:creator>
<dc:creator>Vilankar, K. P.</dc:creator>
<dc:creator>Field, D. J.</dc:creator>
<dc:date>2019-12-28</dc:date>
<dc:identifier>doi:10.1101/2019.12.26.888933</dc:identifier>
<dc:title><![CDATA[Selective and invariant features of neural response surfaces measured with principal curvature]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.30.891549v1?rss=1">
<title>
<![CDATA[
Single cell epigenomic atlas of the developing human brain and organoids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.30.891549v1?rss=1</link>
<description><![CDATA[
Dynamic changes in chromatin accessibility coincide with important aspects of neuronal differentiation, such as fate specification and arealization and confer cell type-specific associations to neurodevelopmental disorders. However, studies of the epigenomic landscape of the developing human brain have yet to be performed at single-cell resolution. Here, we profiled chromatin accessibility of >75,000 cells from eight distinct areas of developing human forebrain using single cell ATAC-seq (scATACseq). We identified thousands of loci that undergo extensive cell type-specific changes in accessibility during corticogenesis. Chromatin state profiling also reveals novel distinctions between neural progenitor cells from different cortical areas not seen in transcriptomic profiles and suggests a role for retinoic acid signaling in cortical arealization. Comparison of the cell type-specific chromatin landscape of cerebral organoids to primary developing cortex found that organoids establish broad cell type-specific enhancer accessibility patterns similar to the developing cortex, but lack many putative regulatory elements identified in homologous primary cell types. Together, our results reveal the important contribution of chromatin state to the emerging patterns of cell type diversity and cell fate specification and provide a blueprint for evaluating the fidelity and robustness of cerebral organoids as a model for cortical development.
]]></description>
<dc:creator>Ziffra, R. S.</dc:creator>
<dc:creator>Kim, C. N.</dc:creator>
<dc:creator>Wilfert, A.</dc:creator>
<dc:creator>Haeussler, M.</dc:creator>
<dc:creator>Casella, A. M.</dc:creator>
<dc:creator>Przytycki, P. F.</dc:creator>
<dc:creator>Kreimer, A.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Ament, S. A.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:date>2019-12-31</dc:date>
<dc:identifier>doi:10.1101/2019.12.30.891549</dc:identifier>
<dc:title><![CDATA[Single cell epigenomic atlas of the developing human brain and organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.01.891432v1?rss=1">
<title>
<![CDATA[
Simultaneous classification of neuroactive compounds in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.01.891432v1?rss=1</link>
<description><![CDATA[
Neuroactive compounds are crucial tools in drug discovery and neuroscience, but it remains difficult to discover neuroactive compounds with new mechanisms of action. To address this need, researchers have developed mid-throughput phenotype-first approaches using zebrafish. This study introduces an open, non-commercial, and extensible hardware/software platform that captures and analyzes drugmodulated phenotypic responses larval zebrafish. We provide full specifications, computer-aided design (CAD) documents, and source code. Accompanying this study, we are also publicly depositing phenotypic data on 3.9 million animals and 34,000 compounds. The data include a high-replicate benchmark set on 14 compounds, a wellcontrolled reference set of 648 known neuroactive compounds, 20 specialized reference sets, a library of 1,520 FDA-approved drugs, 3 screening libraries. This open data resource is curated, structured, tied to extensive metadata, and available under a Creative Commons CC-BY license.
]]></description>
<dc:creator>Myers-Turnbull, D.</dc:creator>
<dc:creator>Taylor, J. C.</dc:creator>
<dc:creator>Helsell, C.</dc:creator>
<dc:creator>Tummino, T. A.</dc:creator>
<dc:creator>Mccarroll, M. N.</dc:creator>
<dc:creator>Alexander, R.</dc:creator>
<dc:creator>Ki, C. S.</dc:creator>
<dc:creator>Gendelev, L.</dc:creator>
<dc:creator>Kokel, D.</dc:creator>
<dc:date>2020-01-02</dc:date>
<dc:identifier>doi:10.1101/2020.01.01.891432</dc:identifier>
<dc:title><![CDATA[Simultaneous classification of neuroactive compounds in zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.01.892281v1?rss=1">
<title>
<![CDATA[
Granular Transcriptomic Signatures Derived from Independent Component Analysis of Bulk Nervous Tissue for Studying Labile Brain Physiologies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.01.892281v1?rss=1</link>
<description><![CDATA[
Multicellular organisms employ concurrent gene regulatory programs to control development and physiology of cells and tissues. The Drosophila melanogaster model system has a remarkable history of revealing the genes and mechanisms underlying fundamental biology yet much remains unclear. In particular, brain xenobiotic protection and endobiotic regulatory systems that require transcriptional coordination across different cell types, operating in parallel with the primary nervous system and metabolic functions of each cell type, are still poorly understood. Here we use the unsupervised machine learning method independent component analysis (ICA) on majority fresh-frozen, bulk tissue microarrays to define biologically pertinent gene expression signatures which are sparse, i.e. each involving only a fraction of all fly genes. We optimize the gene expression signature definitions partly through repeated application of a stochastic ICA algorithm to a compendium of 3,346 microarrays from 221 experiments provided by the Drosophila research community. Our optimized ICA model of pan fly gene expression consists of 850 modules of co-regulated genes that map to tissue developmental stages, disease states, cell-autonomous pathways and presumably novel processes. Importantly, we show biologically relevant gene modules expressed at varying amplitudes in whole brain and isolated adult blood-brain barrier cell levels. Thus, whole tissue derived ICA transcriptional signatures that transcend single cell type boundaries provide a window into the transcriptional states of difficult to isolate cell ensembles maintaining delicate brain physiologies. We believe the fly ICA gene expression signatures set, by virtue of the success of ICA at inferring robust often low amplitude patterns across large datasets and the quality of the input samples, to be an important asset for analyzing compendium and newly generated microarray or RNA-seq expression datasets.
]]></description>
<dc:creator>Rusan, Z. M.</dc:creator>
<dc:creator>Cary, M. P.</dc:creator>
<dc:creator>Bainton, R. J.</dc:creator>
<dc:date>2020-01-02</dc:date>
<dc:identifier>doi:10.1101/2020.01.01.892281</dc:identifier>
<dc:title><![CDATA[Granular Transcriptomic Signatures Derived from Independent Component Analysis of Bulk Nervous Tissue for Studying Labile Brain Physiologies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.05.895227v1?rss=1">
<title>
<![CDATA[
ATP hydrolysis by the SNF2 domain of the ultraspecific maintenance methylase Dnmt5 drives recognition and modification of hemimethylated DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.05.895227v1?rss=1</link>
<description><![CDATA[
C. neoformans Dnmt5 is an ultraspecific maintenance-type CpG methyltransferase (DNMT) that mediates long-term epigenome evolution. It harbors a DNMT domain and SNF2 ATPase domain. We find that the SNF2 domain couples substrate specificity to an ATPase step that is essential for DNA methylation. Such coupling occurs independently of nucleosomes. Hemimethylated DNA preferentially stimulates ATPase activity, and mutating the Dnmt5 ATP binding pocket disproportionately reduces ATPase stimulation by hemimethylated versus unmethylated substrates. Engineered DNA substrates that stabilize a reaction intermediate by mimicking a  flipped-out conformation of the target cytosine bypass the SNF2 domains requirement for hemimethylation. This result implies that ATP hydrolysis by the SNF2 domain is coupled to base-flipping by the DNMT domain. These findings establish a new role for a SNF2 ATPase domain: controlling substrate recognition and catalysis by an adjoined enzymatic domain. This coupling may contribute to the exquisite specificity of Dnmt5 via mechanisms related to kinetic proofreading.
]]></description>
<dc:creator>Dumesic, P. A.</dc:creator>
<dc:creator>Stoddard, C. I.</dc:creator>
<dc:creator>Catania, S.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:creator>Madhani, H.</dc:creator>
<dc:date>2020-01-06</dc:date>
<dc:identifier>doi:10.1101/2020.01.05.895227</dc:identifier>
<dc:title><![CDATA[ATP hydrolysis by the SNF2 domain of the ultraspecific maintenance methylase Dnmt5 drives recognition and modification of hemimethylated DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.07.898023v1?rss=1">
<title>
<![CDATA[
cAMP-Fyn signaling in the dorsomedial striatum direct pathway drives excessive alcohol use 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.07.898023v1?rss=1</link>
<description><![CDATA[
Fyn kinase in the dorsomedial striatum (DMS) of rodents plays a central role in mechanisms underlying excessive alcohol intake. The DMS is comprised of medium spiny neurons (MSNs) that project directly (dMSNs) or indirectly (iMSNs) to the substantia nigra. Here, we examined the cell-type specificity of Fyns actions on alcohol use. First, we knocked down Fyn selectively in DMS dMSNs or iMSNs of mice and measured the level of alcohol consumption. We found that downregulation of Fyn in dMSNs, but not in iMSNs, reduces excessive alcohol but not saccharin intake. D1Rs are coupled to Gs/olf, which activate cAMP signaling. To examine whether Fyns actions are mediated through cAMP signaling, DMS dMSNs were infected with GsDREADD, and the activation of Fyn signaling was measured following CNO treatment. We found that remote stimulation of cAMP signaling in DMS dMSNs activates Fyn and promotes the phosphorylation of the Fyn substrate, GluN2B. In contract, remote activation of GsDREADD in DLS dMSNs did not alter Fyn signaling. We then tested whether activation of GsDREADD in DMS dMSNs or iMSNs alters alcohol intake and observed that CNO-dependent activation of GsDREADD in DMS dMSNs but not iMSNs increases alcohol but not saccharin intake. Finally, we examined the contribution of Fyn to GsDREADD-dependent increase in alcohol intake, and found that systemic administration of the Fyn inhibitor, AZD0503 blocks GsDREADD-dependent increase in alcohol consumption. Our results suggest that the cAMP-Fyn axis in the DMS dMSNs is a molecular transducer of mechanisms underlying the development of excessive alcohol consumption.
]]></description>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>Morisot, N.</dc:creator>
<dc:creator>Sakhai, S.</dc:creator>
<dc:creator>Phamluong, K.</dc:creator>
<dc:creator>Adrover, M.</dc:creator>
<dc:creator>Alvarez, V.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2020-01-08</dc:date>
<dc:identifier>doi:10.1101/2020.01.07.898023</dc:identifier>
<dc:title><![CDATA[cAMP-Fyn signaling in the dorsomedial striatum direct pathway drives excessive alcohol use]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.07.898122v1?rss=1">
<title>
<![CDATA[
ALPPL2 is a highly specific and targetable tumor cell surface antigen 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.07.898122v1?rss=1</link>
<description><![CDATA[
It has been challenging to identify tumor-specific cell surface antigens as the vast majority of tumor-associated antigens are also expressed by some normal tissues. In the course of our study on mesothelioma, we identified a highly specific tumor cell surface antigen that can be targeted for therapy development. Mesothelioma is caused by malignant transformation of the mesothelium, incurable and categorized into three histological subtypes, epithelioid, biphasic and sarcomatoid. To identity novel mesothelioma cell surface antigens with broad subtype coverage and high tissue specificity, we have previously selected phage antibody display libraries on live mesothelioma cells and tissues following counter-selection on normal cells, and identified a panel of human antibodies that bind all subtypes of mesothelioma but not normal mesothelium. One of the antibodies, M25, showed high specificity, and we hereby report the identification of the M25 antigen as ALPPL2. We performed immunohistochemistry on normal human tissues and found that ALPPL2 is expressed only on placental trophoblasts but not any other normal tissues. This exquisite tissue specificity and broad tumor type coverage suggests that ALPPL2 could be an excellent cell surface target for therapeutic development against mesothelioma. To evaluate therapeutic potential of ALPPL2 targeting, we developed an ALPPL2-targeted antibody-drug conjugate and demonstrated potent and specific tumor killing in vitro and in vivo against both epithelioid and sarcomatoid mesothelioma. Thus ALPPL2 belongs to a rare class of cell surface antigens that can be said as being truly tumor specific and is well suited for therapy development against ALPPL2 expressing tumors.
]]></description>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Bidlingmaier, S.</dc:creator>
<dc:creator>Behrens, C. R.</dc:creator>
<dc:creator>Lee, N.-K.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:date>2020-01-08</dc:date>
<dc:identifier>doi:10.1101/2020.01.07.898122</dc:identifier>
<dc:title><![CDATA[ALPPL2 is a highly specific and targetable tumor cell surface antigen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.12.891176v1?rss=1">
<title>
<![CDATA[
Designing and implementing programmable depletion in sequencing libraries with DASHit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.12.891176v1?rss=1</link>
<description><![CDATA[
Since Next-Generation Sequencing produces reads uniformly subsampled from an input library, highly abundant sequences may mask interesting low abundance sequences. The DASH (Depleting Abundant Sequences by Hybridization) technique takes advantage of the programmability of CRISPR/Cas9 to deplete unwanted high-abundance sequences. Because desired depletion targets vary by sample type, here we describe DASHit, software that outputs an optimal DASH target set given a sequencing dataset, an updated DASH protocol, and show depletion results with DASHit-designed targets for three different species.
]]></description>
<dc:creator>Dynerman, D.</dc:creator>
<dc:creator>Lyden, A.</dc:creator>
<dc:creator>Quan, J.</dc:creator>
<dc:creator>Caldera, S.</dc:creator>
<dc:creator>McGeever, A.</dc:creator>
<dc:creator>Dimitrov, B.</dc:creator>
<dc:creator>King, R.</dc:creator>
<dc:creator>Cirola, G.</dc:creator>
<dc:creator>Tan, M.</dc:creator>
<dc:creator>Sit, R.</dc:creator>
<dc:creator>van den Berge, M.</dc:creator>
<dc:creator>Kerstjens, H. A. M.</dc:creator>
<dc:creator>Faiz, A.</dc:creator>
<dc:creator>Christenson, S.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:creator>Crawford, E. D.</dc:creator>
<dc:date>2020-01-13</dc:date>
<dc:identifier>doi:10.1101/2020.01.12.891176</dc:identifier>
<dc:title><![CDATA[Designing and implementing programmable depletion in sequencing libraries with DASHit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.13.905141v1?rss=1">
<title>
<![CDATA[
Recent fluctuations in Mexican American genomes have altered the genetic architecture of biomedical traits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.13.905141v1?rss=1</link>
<description><![CDATA[
People in the Americas represent a diverse group of populations with varying degrees of admixture among African, European, and Amerindigenous ancestries. In the United States, many populations with non-European ancestry remain understudied, and thus little is known about the genetic architecture of phenotypic variation in these populations. Using genome-wide genotype data from the Hispanic Community Health Study/Study of Latinos, we find that Amerindigenous ancestry has increased over time across Hispanic/Latino populations, particularly in Mexican Americans where Amerindigenous ancestry increased by an average of [~]20% over the 50-year period spanning 1940s-1990s. We find similar patterns across American cities, and replicate our observations in an independent sample of Mexican Americans. These dynamic ancestry patterns are a result of a complex interaction of several population and cultural factors, including strong ancestry-related assortative mating and subtle shifts in migration with differences in subcontinental Amerindigenous ancestry over time. These factors have shaped patterns of genetic variation, including an increase in runs of homozygosity in Amerindigenous ancestral tracts, and also influenced the genetic architecture of complex traits within the Mexican American population. We show for height, a trait correlated with ancestry, polygenic risk scores based on summary statistics from a European-based genome-wide association study perform poorly in Mexican Americans. Our findings reveal temporal changes in population structure within Hispanics/Latinos that may influence biomedical traits, demonstrating a crucial need to improve our understanding of the genetic diversity of admixed populations.
]]></description>
<dc:creator>Spear, M. L.</dc:creator>
<dc:creator>Diaz-Papkovich, A.</dc:creator>
<dc:creator>Ziv, E.</dc:creator>
<dc:creator>Gravel, S.</dc:creator>
<dc:creator>Torgerson, D. G.</dc:creator>
<dc:creator>Hernandez, R.</dc:creator>
<dc:date>2020-01-14</dc:date>
<dc:identifier>doi:10.1101/2020.01.13.905141</dc:identifier>
<dc:title><![CDATA[Recent fluctuations in Mexican American genomes have altered the genetic architecture of biomedical traits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.14.906941v1?rss=1">
<title>
<![CDATA[
Clustered γ-Protocadherins Regulate Cortical Interneuron Programmed Cell Death. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.14.906941v1?rss=1</link>
<description><![CDATA[
Cortical function critically depends on inhibitory/excitatory balance. GABAergic cortical inhibitory interneurons (cINs) are born in the ventral forebrain. After completing their migration into cortex, their final numbers are adjusted-during a period of postnatal development - by programmed cell death (PCD). The mechanisms that regulate cIN elimination remain controversial. Here we show that genes in the protocadherin (Pcdh)-{gamma} gene cluster, but not in the Pcdh- or Pcdh-{beta} clusters, are required for survival of cINs through a BAX-dependent mechanism. Surprisingly, the physiological and morphological properties of Pcdh-{gamma} deficient and wild type cINs during PCD were indistinguishable. Co-transplantation of wild type and Pcdh-{gamma} deficient interneuron precursor cells demonstrate that: 1) the number of mutant cINs eliminated was much higher than that of wild type cells, but the proportion of mutant or WT cells undergoing cell death was not affected by their density; 2) the presence of mutant cINs increases cell death among wild-type counterparts, and 3) cIN survival is dependent on the expression of Pcdh-{gamma} C3, C4, and C5. We conclude that Pcdh-{gamma}, and specifically {gamma}C3, {gamma}C4, and {gamma}C5, play a critical role in regulating cIN survival during the endogenous period of PCD.

SignificanceGABAergic cortical inhibitory interneurons (cINs) in the cerebral cortex originate from the ventral embryonic forebrain. After a long migration, they come together with local excitatory neurons to form cortical circuits. These circuits are responsible for higher brain functions, and the improper balance of excitation/inhibition in the cortex can result in mental diseases. Therefore, an understanding of how the final number of cINs is determined is both biologically and, likely, therapeutically significant. Here we show that cell surface homophilic binding proteins belonging to the clustered protocadherin gene family, specifically three isoforms in the Pcdh-{gamma} cluster, play a key role in the regulation cIN programmed cell death. Co-transplantation of mutant and wild-type cINs shows that Pcdh-{gamma} genes have cell-autonomous and non-cell autonomous roles in the regulation of cIN cell death. This work will help identify the molecular mechanisms and cell-cell interactions that determine how the proper ratio of excitatory to inhibitory neurons is determined in the cerebral cortex.
]]></description>
<dc:creator>Mancia Leon, W. R.</dc:creator>
<dc:creator>Spatazza, J.</dc:creator>
<dc:creator>Rakela, B.</dc:creator>
<dc:creator>Chatterjee, A.</dc:creator>
<dc:creator>Pande, V.</dc:creator>
<dc:creator>Maniatis, T.</dc:creator>
<dc:creator>Hasenstaub, A. R.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:date>2020-01-15</dc:date>
<dc:identifier>doi:10.1101/2020.01.14.906941</dc:identifier>
<dc:title><![CDATA[Clustered γ-Protocadherins Regulate Cortical Interneuron Programmed Cell Death.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.15.908152v1?rss=1">
<title>
<![CDATA[
Characterization of Prenylated C-terminal Peptides Using a Novel Capture Technique Coupled with LCMS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.15.908152v1?rss=1</link>
<description><![CDATA[
Post-translational modifications play a critical and diverse role in regulating cellular activities. Despite their fundamentally important role in cellular function, there has been no report to date of an effective generalized approach to the targeting, extraction and characterization of the critical c-terminal regions of natively prenylated proteins. Various chemical modification and metabolic labelling strategies in cell culture have been reported. However, their applicability is limited to cell culture systems and does not allow for analysis of tissue samples. The chemical characteristics (hydrophobicity, low abundance, highly basic charge) of many of the c-terminal regions of prenylated proteins have impaired the use of standard proteomic workflows. In this context, we sought a direct approach to the problem in order to examine these proteins in tissue without the use of labelling. Here we demonstrate that prenylated proteins can be captured on chromatographic resins functionalized with mixed disulfide functions. Protease treatment of resin-bound proteins using chymotryptic digestion revealed peptides from many known prenylated proteins. Exposure of the protease-treated resin to reducing agents and hydro organic mixtures released c-terminal peptides with intact prenyl groups along with other enzymatic modifications expected in this protein family. Database and search parameters were selected to allow for c-terminal modifications unique to these molecules such as CAAX box processing and c-terminal methylation. In summary, we present a direct approach to enrich and obtain information at a molecular level of detail about prenylation of proteins from tissue and cell extracts using high performance LCMS without the need for metabolic labeling and derivatization.



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]]></description>
<dc:creator>Wilkins, J. A.</dc:creator>
<dc:creator>Kaasik, K.</dc:creator>
<dc:creator>Chalkley, R. J.</dc:creator>
<dc:creator>Burlingame, A.</dc:creator>
<dc:date>2020-01-15</dc:date>
<dc:identifier>doi:10.1101/2020.01.15.908152</dc:identifier>
<dc:title><![CDATA[Characterization of Prenylated C-terminal Peptides Using a Novel Capture Technique Coupled with LCMS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.20.911834v1?rss=1">
<title>
<![CDATA[
Loss of Tet2 affects proliferation and drug sensitivity through altered dynamics of cell-state transitions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.20.911834v1?rss=1</link>
<description><![CDATA[
A persistent puzzle in cancer biology is how mutations, which neither alter canonical growth signaling pathways nor directly interfere with drug mechanism, can still recur and persist in tumors. One notable example is the loss-of-function mutation of the DNA demethylase Tet2 in acute myeloid leukemias (AMLs) that frequently persists from diagnosis through remission and relapse (Rothenberg-Thurley et al., 2018; Corces-Zimmerman et al., 2014; Nibourel et al., 2010), but whose fitness advantage in the setting of anti-leukemic chemotherapy is unclear. Here we use paired isogenic human AML cell lines to show that Tet2 loss-of-function alters the dynamics of transitions between differentiated and stem-like states. Mathematical modeling and experimental validation reveal that these altered cell-state dynamics can benefit the cell population by slowing population decay during drug treatment and lowering the number of survivor cells needed to re-establish the initial population. These studies shed light on the functional and phenotypic effects of a Tet2 loss-of-function in AML, illustrate how a single gene mutation can alter a cells phenotypic plasticity, and open up new avenues in the development of strategies to combat AML relapse.
]]></description>
<dc:creator>Morinishi, L. S.</dc:creator>
<dc:creator>Kochanowski, K.</dc:creator>
<dc:creator>Levine, R.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Altschuler, S.</dc:creator>
<dc:date>2020-01-20</dc:date>
<dc:identifier>doi:10.1101/2020.01.20.911834</dc:identifier>
<dc:title><![CDATA[Loss of Tet2 affects proliferation and drug sensitivity through altered dynamics of cell-state transitions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.20.913160v1?rss=1">
<title>
<![CDATA[
Head Movements Control the Activity of Primary Visual Cortex in a Luminance Dependent Manner 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.20.913160v1?rss=1</link>
<description><![CDATA[
The vestibular system broadcasts head-movement related signals to sensory areas throughout the brain, including visual cortex. These signals are crucial for the brains ability to assess whether motion of the visual scene results from the animals head-movements. How head-movements impact visual cortical circuits remains, however, poorly understood. Here, we discover that ambient luminance profoundly transforms how mouse primary visual cortex (V1) processes head-movements. While in darkness, head movements result in an overall suppression of neuronal activity, in ambient light the same head movements trigger excitation across all cortical layers. This light-dependent switch in how V1 processes head-movements is controlled by somatostatin expressing (SOM) inhibitory neurons, which are excited by head movements in dark but not in light. This study thus reveals a light-dependent switch in the response of V1 to head-movements and identifies a circuit in which SOM cells are key integrators of vestibular and luminance signals.
]]></description>
<dc:creator>Bouvier, G.</dc:creator>
<dc:creator>Senzai, Y.</dc:creator>
<dc:creator>Scanziani, M.</dc:creator>
<dc:date>2020-01-21</dc:date>
<dc:identifier>doi:10.1101/2020.01.20.913160</dc:identifier>
<dc:title><![CDATA[Head Movements Control the Activity of Primary Visual Cortex in a Luminance Dependent Manner]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.21.906073v1?rss=1">
<title>
<![CDATA[
Overcoming the inhibitory microenvironment surrounding oligodendrocyte progenitor cells following demyelination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.21.906073v1?rss=1</link>
<description><![CDATA[
Chronic demyelination in the human CNS is characterized by an inhibitory microenvironment that impairs recruitment and differentiation of oligodendrocyte progenitor cells (OPCs) leading to failed remyelination and axonal atrophy. By network-based transcriptomics, we identified sulfatase 2 (Sulf2) mRNA in activated human primary OPCs. Sulf2, an extracellular endosulfatase, modulates the signaling microenvironment by editing the pattern of sulfation on heparan sulfate proteoglycans. We found that Sulf2 was increased in demyelinating lesions in multiple sclerosis and was actively secreted by human OPCs. In experimental demyelination, elevated OPC Sulf1/2 expression directly impaired progenitor recruitment and subsequent generation of oligodendrocytes thereby limiting remyelination. Sulf1/2 potentiates the inhibitory microenvironment by promoting BMP and WNT signaling in OPCs. Importantly, pharmacological sulfatase inhibition using PI-88 accelerated oligodendrocyte recruitment and remyelination by blocking OPC-expressed sulfatases. Our findings define an important inhibitory role of Sulf1/2 and highlight the potential for modulation of the heparanome in the treatment of chronic demyelinating disease.
]]></description>
<dc:creator>Saraswat, D.</dc:creator>
<dc:creator>Polanco, J. J.</dc:creator>
<dc:creator>Shayya, H. J.</dc:creator>
<dc:creator>Tripathi, A.</dc:creator>
<dc:creator>Welliver, R.</dc:creator>
<dc:creator>Pol, S. U.</dc:creator>
<dc:creator>Broome, J. E.</dc:creator>
<dc:creator>O'Bara, M. A.</dc:creator>
<dc:creator>van Kuppervelt, T. H.</dc:creator>
<dc:creator>Phillips, J. J.</dc:creator>
<dc:creator>Dutta, R.</dc:creator>
<dc:creator>Sim, F. J.</dc:creator>
<dc:date>2020-01-21</dc:date>
<dc:identifier>doi:10.1101/2020.01.21.906073</dc:identifier>
<dc:title><![CDATA[Overcoming the inhibitory microenvironment surrounding oligodendrocyte progenitor cells following demyelination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.22.915736v1?rss=1">
<title>
<![CDATA[
Fusion of Drosophila oocytes with specified germline sister cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.22.915736v1?rss=1</link>
<description><![CDATA[
In many animals, oocytes develop together with sister germline cells that pass products to the developing oocyte. In Drosophila, fifteen sister germline (nurse) cells in each egg chamber are known to apoptose by stage 12-13, but we discovered that two specific nurse cells that are juxtaposed to the oocyte are eliminated precociously at stage 10B. These nurse cells fuse with the oocyte and their nuclei extrude through an opening that forms in the oocyte. These nuclei extinguish in the ooplasm, and at stage 11, egg chambers have thirteen nucleated nurse cells and the plasma membrane of the oocyte is mostly restored. In infrequent egg chambers in which nurse cells are not eliminated, oocytes do not develop normally and are not fertilized. Precocious elimination is common to other Drosophila species. We conclude that nurse cells are distinguished by position and identity, and that nurse cell dissolution proceeds in two stages.
]]></description>
<dc:creator>Ali-Murthy, Z.</dc:creator>
<dc:creator>Fetter, R. D.</dc:creator>
<dc:creator>Kornberg, T. B.</dc:creator>
<dc:date>2020-01-23</dc:date>
<dc:identifier>doi:10.1101/2020.01.22.915736</dc:identifier>
<dc:title><![CDATA[Fusion of Drosophila oocytes with specified germline sister cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.22.915926v1?rss=1">
<title>
<![CDATA[
Single cell determination of cardiac microtissue structure and function using light sheet microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.22.915926v1?rss=1</link>
<description><![CDATA[
Native cardiac tissue is comprised of heterogeneous cell populations that work cooperatively for proper tissue function; thus, engineered tissue models have moved toward incorporating multiple cardiac cell types in an effort to recapitulate native multicellular composition and organization. Cardiac tissue models comprised of stem cell-derived cardiomyocytes require inclusion of non-myocytes to promote stable tissue formation, yet the specific contributions of the supporting non-myocyte population on the parenchymal cardiomyocytes and cardiac microtissues have yet to be fully dissected. This gap can be partly attributed to limitations in technologies able to accurately study the individual cellular structure and function that comprise intact 3D tissues. The ability to interrogate the cell-cell interactions in 3D tissue constructs has been restricted by conventional optical imaging techniques that fail to adequately penetrate multicellular microtissues with sufficient spatial resolution. Light sheet fluorescence microscopy overcomes these constraints to enable single cell-resolution structural and functional imaging of intact cardiac microtissues. Multicellular spatial distribution analysis of heterotypic cardiac cell populations revealed that cardiomyocytes and cardiac fibroblasts were randomly distributed throughout 3D microtissues. Furthermore, calcium imaging of live cardiac microtissues enabled single-cell detection of cardiomyocyte calcium activity, which showed that functional heterogeneity correlated with spatial location within the tissues. This study demonstrates that light sheet fluorescence microscopy can be utilized to determine single-cell spatial and functional interactions of multiple cell types within intact 3D engineered microtissues, thereby facilitating the determination of structure-function relationships at both tissue-level and single-cell resolution.

Impact StatementThe ability to achieve single-cell resolution by advanced 3D light imaging techniques enables exquisite new investigation of multicellular analyses in native and engineered tissues. In this study, light sheet fluorescence microscopy was used to define structure-function relationships of distinct cell types in engineered cardiac microtissues by determining heterotypic cell distributions and interactions throughout the tissues as well as by assessing regional differences in calcium handing functional properties at the individual cardiomyocyte level.
]]></description>
<dc:creator>Turaga, D.</dc:creator>
<dc:creator>Matthys, O. B.</dc:creator>
<dc:creator>Hookway, T. A.</dc:creator>
<dc:creator>Joy, D. A.</dc:creator>
<dc:creator>Calvert, M.</dc:creator>
<dc:creator>McDevitt, T. C.</dc:creator>
<dc:date>2020-01-23</dc:date>
<dc:identifier>doi:10.1101/2020.01.22.915926</dc:identifier>
<dc:title><![CDATA[Single cell determination of cardiac microtissue structure and function using light sheet microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.22.916205v1?rss=1">
<title>
<![CDATA[
RBMS1 suppresses colon cancer metastasis through targeted stabilization of its mRNA regulon 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.22.916205v1?rss=1</link>
<description><![CDATA[
Broad dysregulation of gene expression control is a hallmark of cancer progression. Identifying the underlying master regulators that drive pathological gene expression is a key challenge in precision oncology. Here, we have developed a network analytical framework, named PRADA, that identifies oncogenic RNA-binding proteins through the systematic detection of coordinated changes in their target regulons. Application of this approach to data collected from clinical samples, patient-derived xenografts, and cell line models of colon cancer metastasis revealed the RNA-binding protein RBMS1 as a suppressor of colon cancer progression. We observed that silencing RBMS1 results in increased metastatic capacity in xenograft mouse models, and that restoring its expression blunts metastatic liver colonization. We have found that RBMS1 functions as a post-transcriptional regulator of RNA stability by directly binding and stabilizing ~80 target mRNAs. Measurements in more than 180 clinical samples as well as survival analyses in publicly available datasets, have shown that RBMS1 silencing and the subsequent downregulation of its targets are strongly associated with disease progression and poor survival in colon cancer patients. Together, our findings establish a role for RBMS1 as a previously unknown regulator of RNA stability and as a suppressor of colon cancer metastasis with clinical utility for risk stratification of patients.

SignificanceBy applying a new analytical approach to transcriptomic data from clinical samples and models of colon cancer progression, we have uncovered RBMS1 as a suppressor of metastasis and as a post-transcriptional regulator of RNA stability. Notably, RBMS1 silencing and downregulation of its targets are negatively associated with patient survival.
]]></description>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Culbertson, B.</dc:creator>
<dc:creator>Asgharian, H.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Fish, L.</dc:creator>
<dc:creator>Olegario, J. P.</dc:creator>
<dc:creator>Hanisch, B.</dc:creator>
<dc:creator>Weinberg, E.</dc:creator>
<dc:creator>Dienstmann, R.</dc:creator>
<dc:creator>Warren, R.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2020-01-23</dc:date>
<dc:identifier>doi:10.1101/2020.01.22.916205</dc:identifier>
<dc:title><![CDATA[RBMS1 suppresses colon cancer metastasis through targeted stabilization of its mRNA regulon]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.24.919142v1?rss=1">
<title>
<![CDATA[
Neurons in Visual Cortex are Driven by Feedback Projections when their Feedforward Sensory Input is Missing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.24.919142v1?rss=1</link>
<description><![CDATA[
We sense our environment through pathways linking sensory organs to the brain. In the visual system, these feedforward pathways define the classical feedforward receptive field (ffRF), the area in space where visual stimuli excite a neuron1. The visual system also uses visual context, the visual scene surrounding a stimulus, to predict the content of the stimulus2, and accordingly, neurons have been found that are excited by stimuli outside their ffRF3-8. The mechanisms generating excitation to stimuli outside the ffRF are, however, unclear. Here we show that feedback projections onto excitatory neurons in mouse primary visual cortex (V1) generate a second receptive field driven by stimuli outside the ffRF. Stimulating this feedback receptive field (fbRF) elicits slow and delayed responses compared to ffRF stimulation. These responses are preferentially reduced by anesthesia and, importantly, by silencing higher visual areas (HVAs). Feedback inputs from HVAs have scattered receptive fields relative to their putative V1 targets enabling the generation of the fbRF. Neurons with fbRFs are located in cortical layers receiving strong feedback projections and are absent in the main input layer, consistent with a laminar processing hierarchy. The fbRF and the ffRF are mutually antagonistic since large, uniform stimuli, covering both, suppress responses. While somatostatin-expressing inhibitory neurons are driven by these large stimuli, parvalbumin and vasoactive-intestinal-peptide-expressing inhibitory neurons have antagonistic fbRF and ffRF, similar to excitatory neurons. Therefore, feedback projections may enable neurons to use context to predict information missing from the ffRF and to report differences in stimulus features across visual space, regardless if excitation occurs inside or outside the ffRF. We have identified a fbRF which, by complementing the ffRF, may contribute to predictive processing.
]]></description>
<dc:creator>Keller, A. J.</dc:creator>
<dc:creator>Roth, M. M.</dc:creator>
<dc:creator>Scanziani, M.</dc:creator>
<dc:date>2020-01-25</dc:date>
<dc:identifier>doi:10.1101/2020.01.24.919142</dc:identifier>
<dc:title><![CDATA[Neurons in Visual Cortex are Driven by Feedback Projections when their Feedforward Sensory Input is Missing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.24.919191v1?rss=1">
<title>
<![CDATA[
MHC*IMP - Imputation of Alleles for Genes in the Major Histocompatibility Complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.24.919191v1?rss=1</link>
<description><![CDATA[
We report the development of MHC*IMP, a method for imputing non-classical HLA and other genes in the human Major Histocompatibility Complex (MHC). We created a reference panel for 25 genes in the MHC using allele calls from Whole Genome Sequencing data, combined with SNP data for the same individuals. We used this to construct an allele imputation model, MHC*IMP, for each gene. Cross-validation showed that MHC*IMP performs very well, with allele prediction accuracy 93% or greater for all but two of the genes, and greater than 95% for all but four.
]]></description>
<dc:creator>Squire, D. M.</dc:creator>
<dc:creator>Motyer, A.</dc:creator>
<dc:creator>Ahn, R. S.</dc:creator>
<dc:creator>Nitiham, J.</dc:creator>
<dc:creator>Huang, Z.-M.</dc:creator>
<dc:creator>Oksenberg, J. R.</dc:creator>
<dc:creator>Liao, W.</dc:creator>
<dc:creator>Leslie, S.</dc:creator>
<dc:date>2020-01-25</dc:date>
<dc:identifier>doi:10.1101/2020.01.24.919191</dc:identifier>
<dc:title><![CDATA[MHC*IMP - Imputation of Alleles for Genes in the Major Histocompatibility Complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.28.922385v1?rss=1">
<title>
<![CDATA[
Epithelial morphogenesis in the perinatal mouse uterus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.28.922385v1?rss=1</link>
<description><![CDATA[
BackgroundThe uterus is the location where multiple events occur that are required for the start of new life in mammals. The adult uterus contains endometrial or uterine glands that are essential for female fertility. In the mouse, uterine glands are located in the lateral and anti-mesometrial regions of the uterine horn. Previous 3D-imaging of the adult uterus, its glands, and implanting embryos has been performed by multiple groups, using fluorescent microscopy. Adenogenesis, the formation of uterine glands, initiates after birth. Recently, we created a 3D-staging system of mouse uterine gland development at postnatal time points, using light sheet fluorescent microscopy. Here, using a similar approach, we examine the morphological changes in the epithelium of the perinatal mouse uterus.

ResultsThe uterine epithelium exhibits mesometrial-antimesometrial (dorsoventral) patterning as early as three days after birth (P3), marked by the presence of the mesometrially-positioned developing uterine rail. Uterine gland buds are present beginning at P4. Novel morphological epithelial structures, including a ventral ridge and uterine segments were identified.

ConclusionsThe perinatal mouse uterine luminal epithelium develops mesometrial-antimesometrial (dorsal-ventral) morphologies at 3-4 days post-partum. Between 5-6 days post-partum uterine epithelial folds form, defining alternating left-right segments.

Bullet pointsO_LIMorphological patterning events in the perinatal uterine epithelium are not well described.
C_LIO_LILight sheet microscopy was used to generate volumetric reconstructions of the perinatal mouse uterine epithelium.
C_LIO_LIAt postnatal day 3 (P3), the uterine epithelium shows the first signs of dorsoventral pattern, with the presence of the forming mesometrially-positioned uterine rail.
C_LIO_LIThe first morphological indication of uterine adenogenesis begins at P4.
C_LIO_LINovel morphological structures were identified from volumetric reconstructions, including the presence of a ventral ridge (another sign of dorsoventral pattern) and uterine segmentation.
C_LI
]]></description>
<dc:creator>Vue, Z.</dc:creator>
<dc:creator>Behringer, R.</dc:creator>
<dc:date>2020-01-28</dc:date>
<dc:identifier>doi:10.1101/2020.01.28.922385</dc:identifier>
<dc:title><![CDATA[Epithelial morphogenesis in the perinatal mouse uterus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.29.925610v1?rss=1">
<title>
<![CDATA[
Near-atomic structures of the BBSome reveal a novel mechanism for transition zone crossing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.29.925610v1?rss=1</link>
<description><![CDATA[
The BBSome is a complex of eight Bardet-Biedl Syndrome (BBS) proteins that removes signaling receptors from cilia. The GTPase ARL6/BBS3 recruits the BBSome to the ciliary membrane where the BBSome-ARL6GTP complex ferries G protein-coupled receptors (GPCRs) across the transition zone, a diffusion barrier at the base of cilia. Here, we find that the BBSome undergoes a conformational change upon recruitment to membranes by ARL6GTP. Modeling the binding of the BBSome to membranes and to the GPCR Smoothened (SMO) reveals that the amphipathic helix 8 of SMO must be released from the membrane for SMO to be recognized by the BBSome. Underscoring the functional importance of amphipathic helix extraction in TZ crossing, we find that exchanging the amphipathic helix of ARL6 for one that embeds deeper into the membrane blocks BBSome-mediated exit of GPCRs from cilia. We propose that the rigid curvature and dense lipid packing of the transition zone reject asymmetric insertions in the inner leaflet and that the BBSome licenses transition zone crossing by extracting bulky amphipathic helices from the inner leaflet.
]]></description>
<dc:creator>Bahl, K.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Chou, H.-T.</dc:creator>
<dc:creator>Woodsmith, J.</dc:creator>
<dc:creator>Stelzl, U.</dc:creator>
<dc:creator>Walz, T.</dc:creator>
<dc:creator>Nachury, M. V.</dc:creator>
<dc:date>2020-01-30</dc:date>
<dc:identifier>doi:10.1101/2020.01.29.925610</dc:identifier>
<dc:title><![CDATA[Near-atomic structures of the BBSome reveal a novel mechanism for transition zone crossing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.31.929018v1?rss=1">
<title>
<![CDATA[
Cytoprotection by a naturally occurring variant of ATP5G1 in Arctic ground squirrels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.31.929018v1?rss=1</link>
<description><![CDATA[
Many organisms, from anaerobic bacteria to hibernating ground squirrels, have evolved mechanisms to tolerate severe hypoxia or ischemia. In particular, the arctic ground squirrel (AGS) has been shown to be highly resilient to ischemic and reperfusion injuries, demonstrating an ability to withstand metabolic stress under hibernation conditions. Although physiological adaptations are critical to ischemic tolerance in AGS, little is known about cellular mechanisms underlying intrinsic AGS cell tolerance to metabolic stressors. Through cell survival-based cDNA expression screens and comparative genomics, we have discovered that in AGS, a cytoprotective variant of ATP5G1 helps confer improved mitochondrial metabolism and cell resilience to metabolic stress. ATP5G1 encodes a proton-transporting subunit of the mitochondrial ATP synthase complex. Ectopic expression in mouse cells and CRISPR/Cas9 base editing of the endogenous AGS locus revealed causal roles of one AGS-specific amino acid substitution (leucine-32) in mediating the cytoprotective effects of AGS ATP5G1. We provide evidence that AGS ATP5G1 promotes cell resilience to stress by modulating mitochondrial morphological change and metabolic functions. Thus, our results identify a naturally occurring variant of ATP5G1 from a mammalian hibernator that causally contributes to intrinsic cytoprotection against metabolic stresses.
]]></description>
<dc:creator>Singhal, N. S.</dc:creator>
<dc:creator>Bai, M.</dc:creator>
<dc:creator>Lee, E. M.</dc:creator>
<dc:creator>Luo, S.</dc:creator>
<dc:creator>Cook, K. R.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2020-01-31</dc:date>
<dc:identifier>doi:10.1101/2020.01.31.929018</dc:identifier>
<dc:title><![CDATA[Cytoprotection by a naturally occurring variant of ATP5G1 in Arctic ground squirrels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.31.929166v1?rss=1">
<title>
<![CDATA[
A Disinhibitory Circuit for Contextual Modulation in Primary Visual Cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.31.929166v1?rss=1</link>
<description><![CDATA[
Context guides perception by influencing the saliency of sensory stimuli. Accordingly, in visual cortex, responses to a stimulus are modulated by context, the visual scene surrounding the stimulus. Responses are suppressed when stimulus and surround are similar but not when they differ. The mechanisms that remove suppression when stimulus and surround differ remain unclear. Here we use optical recordings, manipulations, and computational modelling to show that a disinhibitory circuit consisting of vasoactive-intestinal-peptide-expressing (VIP) and somatostatin-expressing (SOM) inhibitory neurons modulates responses in mouse visual cortex depending on the similarity between stimulus and surround. When the stimulus and the surround are similar, VIP neurons are inactive and SOM neurons suppress excitatory neurons. However, when the stimulus and the surround differ, VIP neurons are active, thereby inhibiting SOM neurons and relieving excitatory neurons from suppression. We have identified a canonical cortical disinhibitory circuit which contributes to contextual modulation and may regulate perceptual saliency.
]]></description>
<dc:creator>Keller, A. J.</dc:creator>
<dc:creator>Roth, M. M.</dc:creator>
<dc:creator>Caudill, M.</dc:creator>
<dc:creator>Dipoppa, M.</dc:creator>
<dc:creator>Miller, K. D.</dc:creator>
<dc:creator>Scanziani, M.</dc:creator>
<dc:date>2020-01-31</dc:date>
<dc:identifier>doi:10.1101/2020.01.31.929166</dc:identifier>
<dc:title><![CDATA[A Disinhibitory Circuit for Contextual Modulation in Primary Visual Cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.03.927442v1?rss=1">
<title>
<![CDATA[
Poecivirus is present in individuals with beak deformities in seven species of North American birds 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.03.927442v1?rss=1</link>
<description><![CDATA[
Avian keratin disorder (AKD), a disease characterized by debilitating beak overgrowth but with unknown etiology, has increasingly affected wild bird populations since the 1990s. We previously showed that a novel picornavirus, Poecivirus, is closely correlated with disease status in Black-capped Chickadees (Poecile atricapillus) in Alaska. However, our knowledge of the relationship between Poecivirus and beak deformities in other species and other geographic areas remains limited. The growing geographic scope and number of species affected by AKD-like beak deformities require a better understanding of the causative agent to evaluate the population-level impacts of this epizootic. Here, we tested eight individuals from six avian species with AKD-consistent deformities for the presence of Poecivirus: Mew Gull (Larus canus), Hairy Woodpecker (Picoides villosus), Black-billed Magpie (Pica hudsonia), Northwestern Crow (Corvus caurinus), Red-breasted Nuthatch (Sitta canadensis), and Blackpoll Warbler (Setophaga striata); individuals were sampled in Alaska and Maine (1999-2016). We used targeted PCR followed by Sanger sequencing to test for the presence of Poecivirus in each specimen, and to obtain viral genome sequence from virus-positive host individuals. We detected Poecivirus in all individuals tested, but not in negative controls. Furthermore, we used unbiased metagenomic sequencing to test for the presence of other pathogens in six of these specimens (Hairy Woodpecker, two Northwestern Crows, two Red-breasted Nuthatches, Blackpoll Warbler). This analysis yielded additional viral sequence from several specimens, including the complete coding region of Poecivirus from one Red-breasted Nuthatch, which we confirmed via targeted PCR followed by Sanger sequencing. This study demonstrates that Poecivirus is present in individuals with AKD-consistent deformities from six avian species other than Black-capped Chickadee. While further investigation will be required to explore whether there exists a causal link between this virus and AKD, this study demonstrates that Poecivirus is not geographically restricted to Alaska, but rather occurs elsewhere in North America.
]]></description>
<dc:creator>Zylberberg, M.</dc:creator>
<dc:creator>Van Hemert, C.</dc:creator>
<dc:creator>Handel, C. M.</dc:creator>
<dc:creator>Liu, R.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:date>2020-02-04</dc:date>
<dc:identifier>doi:10.1101/2020.02.03.927442</dc:identifier>
<dc:title><![CDATA[Poecivirus is present in individuals with beak deformities in seven species of North American birds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.04.932988v1?rss=1">
<title>
<![CDATA[
ZipSeq : Barcoding for Real-time Mapping of Single Cell Transcriptomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.04.932988v1?rss=1</link>
<description><![CDATA[
Spatial transcriptomics seeks to integrate single-cell transcriptomic data within the 3-dimensional space of multicellular biology. Current methods use glass substrates pre-seeded with matrices of barcodes or fluorescence hybridization of a limited number of probes. We developed an alternative approach, called  ZipSeq, that uses patterned illumination and photocaged oligonucleotides to serially print barcodes (Zipcodes) onto live cells within intact tissues, in real-time and with on-the-fly selection of patterns. Using ZipSeq, we mapped gene expression in three settings: in-vitro wound healing, live lymph node sections and in a live tumor microenvironment (TME). In all cases, we discovered new gene expression patterns associated with histological structures. In the TME, this demonstrated a trajectory of myeloid and T cell differentiation, from periphery inward. A variation of ZipSeq efficiently scales to the level of single cells, providing a pathway for complete mapping of live tissues, subsequent to real-time imaging or perturbation.
]]></description>
<dc:creator>Hu, K. H.</dc:creator>
<dc:creator>Eichorst, J. P.</dc:creator>
<dc:creator>McGinnis, C. S.</dc:creator>
<dc:creator>Patterson, D. M.</dc:creator>
<dc:creator>Chow, E. D.</dc:creator>
<dc:creator>Kersten, K.</dc:creator>
<dc:creator>Jameson, S. C.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:creator>Rao, A. A.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2020-02-04</dc:date>
<dc:identifier>doi:10.1101/2020.02.04.932988</dc:identifier>
<dc:title><![CDATA[ZipSeq : Barcoding for Real-time Mapping of Single Cell Transcriptomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.05.936195v1?rss=1">
<title>
<![CDATA[
Principles of dengue virus evolvability derived from genotype-fitness maps in human and mosquito cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.05.936195v1?rss=1</link>
<description><![CDATA[
Dengue virus (DENV), an arbovirus infecting over 100 million annually, cycles between human and mosquito hosts1. Examining how DENV adapts to such different host environments could uncover principles of arbovirus transmission and emergence. Here we combine sequential passaging and ultra-deep sequencing to examine the sequence dynamics and fitness changes of DENV populations adapting to human and mosquito cells, identifying the contributions of beneficial and deleterious mutations in shaping the fitness landscape driving host-specific paths of viral adaptation. We find DENV phenotypic adaptation is best described by the collective fitness contributions of all the alleles present in the population. Accordingly, while increased fitness during adaptation to each host is driven by host-specific beneficial mutations, it is reduced by the consistently replenished genetic load of deleterious mutations. Of note, host-specific beneficial mutations are in discrete regions across the genome, revealing molecular mechanisms of adaptation. Some of these clusters comprise phenotypically redundant mutations that may provide evolutionary robustness to transmission bottlenecks. Our results also suggest DENV adaptation is facilitated through variation in intrinsically disordered protein regions while transmembrane and structured domains evolve under stronger biophysical constraints. Importantly, the adaptation strategies uncovered in our simple system mirror macro-evolutionary changes observed across DENV serotypes and Zika virus and may suggest general principles of evolvability in arbovirus evolution.
]]></description>
<dc:creator>Dolan, P. T.</dc:creator>
<dc:creator>Taguwa, S.</dc:creator>
<dc:creator>Aguilar Rangel, M.</dc:creator>
<dc:creator>Acevedo, A.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:date>2020-02-07</dc:date>
<dc:identifier>doi:10.1101/2020.02.05.936195</dc:identifier>
<dc:title><![CDATA[Principles of dengue virus evolvability derived from genotype-fitness maps in human and mosquito cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.06.936070v1?rss=1">
<title>
<![CDATA[
MicroRNA-dependent inhibition of PFN2 orchestrates ERK activation and pluripotent state transitions by regulating endocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.06.936070v1?rss=1</link>
<description><![CDATA[
Profilin2 (PFN2) is a target of the embryonic stem cell (ESC) enriched miR-290 family of miRNAs and an actin/dynamin binding protein implicated in endocytosis. Here, we show that the miR-290-PFN2 pathway regulates many aspects of ESC biology. In the absence of microRNAs, PFN2 is upregulated in ESCs, with a resulting decrease in endocytosis. Reintroduction of miR-290, knockout of PFN2, or disruption of the PFN2 dynamin interacting domain in miRNA deficient cells reverses the endocytosis defect. The loss of miRNA suppression of PFN2 and associated reduction in endocytosis impairs ERK signaling, which in turn inhibits ESC cell cycle progression and differentiation from a naive to formative state. Mutagenesis of the single canonical conserved 3UTR miR-290 binding site of PFN2 in otherwise wild-type cells recapitulates these phenotypes. Together, these findings define an axis of post-transcriptional control, endocytosis, and signal transduction that is essential for ESC self-renewal and differentiation.
]]></description>
<dc:creator>Sangokoya, C.</dc:creator>
<dc:creator>Blelloch, R.</dc:creator>
<dc:date>2020-02-06</dc:date>
<dc:identifier>doi:10.1101/2020.02.06.936070</dc:identifier>
<dc:title><![CDATA[MicroRNA-dependent inhibition of PFN2 orchestrates ERK activation and pluripotent state transitions by regulating endocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.09.940882v1?rss=1">
<title>
<![CDATA[
BET Bromodomain Proteins Regulate Transcriptional Reprogramming in Genetic Dilated Cardiomyopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.09.940882v1?rss=1</link>
<description><![CDATA[
The bromodomain and extraterminal (BET) family of epigenetic reader proteins are key regulators of pathologic gene expression in the heart. Using mice carrying a human mutation in phospholamban (PLNR9C) that develop progressive dilated cardiomyopathy (DCM), we previously identified the activation of inflammatory gene networks as a key early driver of DCM. We reasoned that BETs control this inflammatory process, representing a key node in the progression of genetic DCM. Using a chemical genetic strategy, PLNR9C or age-matched wild type mice were treated longitudinally with the BET inhibitor JQ1 or vehicle. JQ1 abrogated DCM, reduced cardiac fibrosis, and prolonged survival in PLNR9C mice by inhibiting inflammatory gene network expression at all disease stages. Cardiac fibroblast proliferation was also substantially reduced by JQ1. Interestingly, JQ1 had profound effects on pathologic gene network expression in cardiac fibroblasts, while having little effect on transcription in cardiomyocytes. Using co-immunoprecipitation, we identified BRD4 as a direct and essential regulator of NF{kappa}B-mediated inflammatory gene transcription in cardiac fibroblasts. In this this model of chronic, heritable DCM, BETs activate inflammatory gene networks in cardiac fibroblasts via an NF{kappa}B-dependent mechanism, marking them as critical effectors of pathologic gene expression.
]]></description>
<dc:creator>Antolic, A.</dc:creator>
<dc:creator>Wakimoto, H.</dc:creator>
<dc:creator>Jiao, Z.</dc:creator>
<dc:creator>Gorham, J. M.</dc:creator>
<dc:creator>DePalma, S. R.</dc:creator>
<dc:creator>Conner, D. A.</dc:creator>
<dc:creator>Lee, D. Y.</dc:creator>
<dc:creator>Qi, J.</dc:creator>
<dc:creator>Seidman, J. G.</dc:creator>
<dc:creator>Bradner, J. E.</dc:creator>
<dc:creator>Brown, J. D.</dc:creator>
<dc:creator>Haldar, S. M.</dc:creator>
<dc:creator>Seidman, C. E.</dc:creator>
<dc:creator>Burke, M. A.</dc:creator>
<dc:date>2020-02-10</dc:date>
<dc:identifier>doi:10.1101/2020.02.09.940882</dc:identifier>
<dc:title><![CDATA[BET Bromodomain Proteins Regulate Transcriptional Reprogramming in Genetic Dilated Cardiomyopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.12.945063v1?rss=1">
<title>
<![CDATA[
Massively Parallel Analysis of Human 3' UTRs Reveals that AU-Rich Element Length and Registration Predict mRNA Destabilization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.12.945063v1?rss=1</link>
<description><![CDATA[
AU-rich elements (AREs) are 3' UTR cis-regulatory elements that regulate the stability of mRNAs. Consensus ARE motifs have been determined, but little is known about how differences in 3' UTR sequences that conform to these motifs affect their function. Here we use functional annotation of sequences from 3' UTRs (fast-UTR), a massively parallel reporter assay (MPRA), to investigate the effects of 41,288 3' UTR sequence fragments from 4,653 transcripts on gene expression and mRNA stability. The library included 9,142 AREs, and incorporated a set of fragments bearing mutations in each ARE. Our analyses demonstrate that the length of an ARE and its registration (the first and last nucleotides of the repeating ARE motif) have significant effects on gene expression and stability. Based on this finding, we propose improved ARE classification and concomitant methods to categorize and predict the effect of AREs on gene expression and stability. Our new approach explains 64{+/-}13% of the contribution of AREs to the stability of human 3' UTRs in Jurkat cells and predicts ARE activity in an unrelated cell type. Finally, to investigate the advantages of our general experimental design for annotating 3' UTR elements we examine other motifs including constitutive decay elements (CDEs), where we show that the length of the CDE stem-loop has a significant impact on steady-state expression and mRNA stability. We conclude that fast-UTR, in conjunction with our analytical approach, can produce improved yet simple sequence-based rules for predicting the activity of human 3' UTRs containing functional motifs.
]]></description>
<dc:creator>Siegel, D. A.</dc:creator>
<dc:creator>Le Tonqueze, O.</dc:creator>
<dc:creator>Biton, A.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:date>2020-02-12</dc:date>
<dc:identifier>doi:10.1101/2020.02.12.945063</dc:identifier>
<dc:title><![CDATA[Massively Parallel Analysis of Human 3' UTRs Reveals that AU-Rich Element Length and Registration Predict mRNA Destabilization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.16.951772v1?rss=1">
<title>
<![CDATA[
High-temperature live-cell imaging of cytokinesis, cell motility and cell-cell adhesion in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.16.951772v1?rss=1</link>
<description><![CDATA[
Significant technical challenges have limited the study of extremophile cell biology. For example, the absence of methods for performing high-resolution, live-cell imaging at high temperatures (>50{degrees}C) has impeded the study of cell motility and cell division in thermophilic archaea such as model organisms from the genus Sulfolobus. Here we describe a system for imaging samples at 75{degrees}C using high numerical aperture, oil-immersion lenses. With this system we observed and quantified the dynamics of cell division in the model thermoacidophilic crenarchaeon Sulfolobus acidocaldarius. In addition, we observed previously undescribed dynamic cell shape changes, cell motility, and cell-cell interactions, shedding significant new light on the high-temperature lifestyle of this organism.
]]></description>
<dc:creator>Charles-Orszag, A.</dc:creator>
<dc:creator>Lord, S. J.</dc:creator>
<dc:creator>Mullins, D.</dc:creator>
<dc:date>2020-02-17</dc:date>
<dc:identifier>doi:10.1101/2020.02.16.951772</dc:identifier>
<dc:title><![CDATA[High-temperature live-cell imaging of cytokinesis, cell motility and cell-cell adhesion in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.17.952093v1?rss=1">
<title>
<![CDATA[
Oncogenic mutant RAS activity is rescaled by the ERK/MAPK pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.17.952093v1?rss=1</link>
<description><![CDATA[
Activating mutations in RAS are present in [~]30% of human tumors, and the resulting aberrations in ERK/MAPK signaling play a central role in oncogenesis. However, the form of these signaling changes is uncertain, with activating RAS mutants linked to both increased and decreased ERK activation in vivo. Rationally targeting the kinase activity of this pathway requires clarification of the quantitative effects of RAS mutations. Here, we use live-cell imaging in cell lines expressing only one RAS isoform to quantify ERK activity with a new level of accuracy. We find that despite large differences in their biochemical activity, mutant KRAS isoforms within cells have similar ranges of ERK output. We identify roles for pathway-level effects, including variation in feedback strength and feedforward modulation of phosphatase activity, that act to rescale pathway sensitivity independent of expression level, ultimately resisting changes in the dynamic range of ERK activity while preserving responsiveness to growth factor stimuli. Our results reconcile seemingly inconsistent reports within the literature and imply that the initial signaling changes induced by RAS mutations in oncogenesis are subtle.
]]></description>
<dc:creator>Gillies, T. E.</dc:creator>
<dc:creator>Pargett, M.</dc:creator>
<dc:creator>Silva, J. M.</dc:creator>
<dc:creator>Teragawa, C.</dc:creator>
<dc:creator>McCormick, F.</dc:creator>
<dc:creator>Albeck, J. G.</dc:creator>
<dc:date>2020-02-17</dc:date>
<dc:identifier>doi:10.1101/2020.02.17.952093</dc:identifier>
<dc:title><![CDATA[Oncogenic mutant RAS activity is rescaled by the ERK/MAPK pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.24.962688v1?rss=1">
<title>
<![CDATA[
IFN-{lambda}4 increases the risk of gastrointestinal infections and malaria in Malian children 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.24.962688v1?rss=1</link>
<description><![CDATA[
Genetic polymorphisms within the IFNL3/IFNL4 genomic region, which encodes type III interferons, have been strongly associated with impaired clearance of hepatitis C virus (HCV) infection. We hypothesized that type III interferons might be important for the immune response to other pathogens as well. In a cohort of 914 Malian children, we analyzed episodes of malaria, gastrointestinal and respiratory infections using information for 30,626 clinic visits from birth through up to 5 years of follow-up. Genetic polymorphisms IFNL4-rs368234815 and IFNL3-rs4803217 that functionally affect type III interferons were genotyped with TaqMan assays. For both genetic variants and each infection, we evaluated time-to-first episode and calculated odds ratios (ORs) for the risk of an infection episode during follow-up, controlling for relevant covariates. Compared to children with the rs368234815-TT/TT genotype (IFN-{lambda}4-Null), each copy of the rs368234815-dG allele was associated with an earlier first episode of a gastrointestinal infection (p=0.003) and respiratory infection (p=0.045). The risk of experiencing an infection episode during the follow-up was also significantly increased with each copy of the rs368234815-dG allele - for gastrointestinal infections (OR=1.53, 95%CI (1.13-2.07), p=0.005) and malaria (OR=1.30, 95%CI (1.02-1.65), p=0.033). IFNL4-rs368234815 and IFNL3-rs4803217 were in moderate linkage disequilibrium in this population (r2=0.78), and all the associations for rs4803217 were weaker and lost significance after adjusting for rs368234815, implicating IFN-{lambda}4 and not IFN-{lambda}3 as the primary cause of these associations. We conclude that the ability to produce IFN-{lambda}4 may have broad health-related implications by negatively affecting the immune response and clinical outcomes of several common infections.
]]></description>
<dc:creator>Prokunina-Olsson, L.</dc:creator>
<dc:creator>Morrison, R. L.</dc:creator>
<dc:creator>Obajemu, A.</dc:creator>
<dc:creator>Mahamar, A.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Attaher, O.</dc:creator>
<dc:creator>Florez-Vargas, O.</dc:creator>
<dc:creator>Sidibe, Y.</dc:creator>
<dc:creator>Onabajo, O. O.</dc:creator>
<dc:creator>Hutchinson, A. A.</dc:creator>
<dc:creator>Manning, M.</dc:creator>
<dc:creator>Kwan, J.</dc:creator>
<dc:creator>Brand, N.</dc:creator>
<dc:creator>Dicko, A.</dc:creator>
<dc:creator>Fried, M.</dc:creator>
<dc:creator>Albert, P. S.</dc:creator>
<dc:creator>Mbulaiteye, S. M.</dc:creator>
<dc:creator>Duffy, P. E.</dc:creator>
<dc:date>2020-02-25</dc:date>
<dc:identifier>doi:10.1101/2020.02.24.962688</dc:identifier>
<dc:title><![CDATA[IFN-{lambda}4 increases the risk of gastrointestinal infections and malaria in Malian children]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.26.966911v1?rss=1">
<title>
<![CDATA[
CRISPR Screen in Regulatory T Cells Reveals Ubiquitination Modulators of Foxp3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.26.966911v1?rss=1</link>
<description><![CDATA[
Regulatory T cells (Tregs) are required to control immune responses and maintain homeostasis but are a significant barrier to anti-tumor immunity1. Conversely, Treg instability, characterized by loss of the master transcription factor Foxp3 and acquisition of pro-inflammatory properties2, can promote autoimmunity and/or facilitate more effective tumor immunity3,4. A comprehensive understanding of the pathways that regulate Foxp3 could lead to more effective Treg therapies for autoimmune disease and cancer. Despite improved functional genetic tools that now allow for systematic interrogation, dissection of the gene regulatory programs that modulate Foxp3 expression has not yet been reported. In this study, we developed a CRISPR-based pooled screening platform for phenotypes in primary mouse Tregs and applied this technology to perform a targeted loss-of-function screen of [~]490 nuclear factors to identify gene regulatory programs that promote or disrupt Foxp3 expression. We discovered several novel modulators including ubiquitin-specific peptidase 22 (Usp22), Ataxin 7 like 3 (Atxn7l3) and ring finger protein 20 (Rnf20). Members of the deubiquitination module of the SAGA chromatin modifying complex, Usp22 and Atxn7l3, were discovered to be positive regulators that stabilized Foxp3 expression; whereas the screen suggested Rnf20, an E3 ubiquitin ligase, is a negative regulator of Foxp3. Treg-specific ablation of Usp22 in mice reduced Foxp3 protein and created defects in their suppressive function that led to spontaneous autoimmunity but protected against tumor growth in multiple cancer models. Foxp3 destabilization in Usp22-deficient Tregs could be rescued by ablation of Rnf20, revealing a reciprocal ubiquitin switch in Tregs. These results reveal novel modulators of Foxp3 and demonstrate a screening method that can be broadly applied to discover new targets for Treg immunotherapies for cancer and autoimmune disease.
]]></description>
<dc:creator>Cortez, J. T.</dc:creator>
<dc:creator>Montauti, E.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Shaked, O.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Roth, T.</dc:creator>
<dc:creator>Simeonov, D.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Vogel, I. A.</dc:creator>
<dc:creator>Prator, G. Y.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Lee, Y.</dc:creator>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Ifergan, I.</dc:creator>
<dc:creator>Van Gool, F.</dc:creator>
<dc:creator>Bluestone, J. A.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Fang, D.</dc:creator>
<dc:date>2020-02-27</dc:date>
<dc:identifier>doi:10.1101/2020.02.26.966911</dc:identifier>
<dc:title><![CDATA[CRISPR Screen in Regulatory T Cells Reveals Ubiquitination Modulators of Foxp3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.27.968883v1?rss=1">
<title>
<![CDATA[
Molecular goniometers for single-particle cryo-EM of DNA-binding proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.27.968883v1?rss=1</link>
<description><![CDATA[
Correct reconstruction of macromolecular structure by cryo-electron microscopy relies on accurate determination of the orientation of single-particle images. For small (<100 kDa) DNA-binding proteins, obtaining particle images with sufficiently asymmetric features to correctly guide alignment is challenging. DNA nanotechnology was conceived as a potential tool for building host nanostructures to prescribe the locations and orientations of docked proteins. We used DNA origami to construct molecular goniometers--instruments to precisely orient objects--to dock a DNA-binding protein on a double-helix stage that has user-programmable tilt and rotation angles. Each protein orientation maps to a distinct barcode pattern specifying particle classification and angle assignment. We used goniometers to obtain a 6.5 [A] structure of BurrH, an 82-kDa DNA-binding protein whose helical pseudosymmetry prevents accurate image orientation using classical cryo-EM. Our approach should be adaptable for other DNA-binding proteins, and a wide variety of other small proteins, by fusing DNA binding domains to them.
]]></description>
<dc:creator>Aksel, T.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Douglas, S. M.</dc:creator>
<dc:date>2020-02-28</dc:date>
<dc:identifier>doi:10.1101/2020.02.27.968883</dc:identifier>
<dc:title><![CDATA[Molecular goniometers for single-particle cryo-EM of DNA-binding proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.29.970558v1?rss=1">
<title>
<![CDATA[
An integrated transcriptomic and epigenomic atlas of mouse primary motor cortex cell types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.29.970558v1?rss=1</link>
<description><![CDATA[
Single cell transcriptomics has transformed the characterization of brain cell identity by providing quantitative molecular signatures for large, unbiased samples of brain cell populations. With the proliferation of taxonomies based on individual datasets, a major challenge is to integrate and validate results toward defining biologically meaningful cell types. We used a battery of single-cell transcriptome and epigenome measurements generated by the BRAIN Initiative Cell Census Network (BICCN) to comprehensively assess the molecular signatures of cell types in the mouse primary motor cortex (MOp). We further developed computational and statistical methods to integrate these multimodal data and quantitatively validate the reproducibility of the cell types. The reference atlas, based on more than 600,000 high quality single-cell or -nucleus samples assayed by six molecular modalities, is a comprehensive molecular account of the diverse neuronal and non-neuronal cell types in MOp. Collectively, our study indicates that the mouse primary motor cortex contains over 55 neuronal cell types that are highly replicable across analysis methods, sequencing technologies, and modalities. We find many concordant multimodal markers for each cell type, as well as thousands of genes and gene regulatory elements with discrepant transcriptomic and epigenomic signatures. These data highlight the complex molecular regulation of brain cell types and will directly enable design of reagents to target specific MOp cell types for functional analysis.
]]></description>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Xie, F.</dc:creator>
<dc:creator>Fischer, S.</dc:creator>
<dc:creator>Adkins, R. S.</dc:creator>
<dc:creator>Aldrige, A. I.</dc:creator>
<dc:creator>Ament, S. A.</dc:creator>
<dc:creator>Bartlett, A.</dc:creator>
<dc:creator>Behrens, M. M.</dc:creator>
<dc:creator>Van den Berge, K.</dc:creator>
<dc:creator>Bertagnolli, D.</dc:creator>
<dc:creator>Biancalani, T.</dc:creator>
<dc:creator>Booeshaghi, A. S.</dc:creator>
<dc:creator>Corrada Bravo, H.</dc:creator>
<dc:creator>Casper, T.</dc:creator>
<dc:creator>Colantuoni, C.</dc:creator>
<dc:creator>Crabtree, J.</dc:creator>
<dc:creator>Creasy, H.</dc:creator>
<dc:creator>Crichton, K.</dc:creator>
<dc:creator>Crow, M.</dc:creator>
<dc:creator>Dee, N.</dc:creator>
<dc:creator>Dougherty, E. L.</dc:creator>
<dc:creator>Doyle, W. I.</dc:creator>
<dc:creator>Dudoit, S.</dc:creator>
<dc:creator>Fang, R.</dc:creator>
<dc:creator>Felix, V.</dc:creator>
<dc:creator>Fong, O.</dc:creator>
<dc:creator>Giglio, M.</dc:creator>
<dc:creator>Goldy, J.</dc:creator>
<dc:creator>Hawrylycz, M.</dc:creator>
<dc:creator>Bezieux, H. R. d.</dc:creator>
<dc:creator>Herb, B. R.</dc:creator>
<dc:creator>Hertzano, R.</dc:creator>
<dc:creator>Hou, X.</dc:creator>
<dc:creator>Hu, Q.</dc:creator>
<dc:creator>Huang, Z. J.</dc:creator>
<dc:creator>Kancherla, J.</dc:creator>
<dc:creator>Kroll, M.</dc:creator>
<dc:creator>Lathia, K.</dc:creator>
<dc:creator>Li, Y. E.</dc:creator>
<dc:creator>Lucero, J. D.</dc:creator>
<dc:creator>Luo, C.</dc:creator>
<dc:creator>Mahurkar, A.</dc:creator>
<dc:creator>McMillen, D.</dc:creator>
<dc:creator>Nadaf, N. M.</dc:creator>
<dc:creator>Ner</dc:creator>
<dc:date>2020-03-02</dc:date>
<dc:identifier>doi:10.1101/2020.02.29.970558</dc:identifier>
<dc:title><![CDATA[An integrated transcriptomic and epigenomic atlas of mouse primary motor cortex cell types]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.02.972745v1?rss=1">
<title>
<![CDATA[
Fusion pore expansion and contraction during catecholamine release from endocrine cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.02.972745v1?rss=1</link>
<description><![CDATA[
Amperometry recording reveals the exocytosis of catecholamine from individual vesicles as a sequential process, typically beginning slowly with a pre-spike foot, accelerating sharply to initiate a spike, reaching a peak, and then decaying. This complex sequence reflects the interplay between diffusion, flux through a fusion pore, and possibly dissociation from a vesicles densecore. In an effort to evaluate the impacts of these factors, a model was developed that combines diffusion with flux through a static pore. This model recapitulated the rapid phases of a spike, but generated relations between spike shape parameters that differed from experimental results. To explore the possibility of fusion pore dynamics, a transformation of amperometry current was introduced that yields fusion pore permeability divided by vesicle volume (g/V). Applying this transform to individual fusion events yielded a highly characteristic time course. g/V initially tracks the pre-spike foot and the start of the spike, increasing [~]15-fold to the peak. However, after the spike peaks, g/V unexpectedly declines and settles to a constant value that indicates the presence of a stable post-spike pore. g/V of the post-spike pore varies greatly between events, and has an average that is [~]3.5-fold below the peak value and [~]4.5-fold above the pre-spike value. The post-spike pore persists and g/V remains flat for tens of milliseconds, as long as catecholamine flux can be detected. Applying the g/V transform to rare events with two peaks revealed a stepwise increase in g/V during the second peak. The g/V transform offers an interpretation of amperometric current in terms of fusion pore dynamics and provides a new framework for analyzing the actions of proteins that alter spike shape. The stable post-spike pore conforms with predictions from lipid bilayer elasticity, and offers an explanation for previous reports of prolonged hormone retention within fusing vesicles.

STATEMENT OF SIGNIFICANCEAmperometry recordings of catecholamine release from single vesicles reveal a complex waveform with distinct phases. The role of the fusion pore in this waveform is poorly understood. A model based on a static fusion pore fails to recapitulate important aspects of the waveform. A new transform of amperometric current introduced here renders fusion pore permeability in real time. This transform reveals rich dynamic behavior of the fusion pore as catecholamine leaves a vesicle. This analysis shows that fusion pore permeability rapidly increases and then decreases before settling into a stable post-spike configuration.
]]></description>
<dc:creator>Jackson, M. B.</dc:creator>
<dc:creator>Hsiao, Y.-T.</dc:creator>
<dc:creator>Chang, C.-W.</dc:creator>
<dc:date>2020-03-02</dc:date>
<dc:identifier>doi:10.1101/2020.03.02.972745</dc:identifier>
<dc:title><![CDATA[Fusion pore expansion and contraction during catecholamine release from endocrine cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.04.977090v1?rss=1">
<title>
<![CDATA[
Lysine63-linked ubiquitin chains earmark GPCRs for BBSome-mediated removal from cilia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.04.977090v1?rss=1</link>
<description><![CDATA[
Regulated trafficking of G-protein coupled receptors (GPCRs) controls cilium-based signaling pathways. {beta}-arrestin, a molecular sensor of activated GPCRs, and the BBSome, a complex of Bardet-Biedl Syndrome (BBS) proteins, are required for the signal-dependent exit of ciliary GPCRs but the functional interplay between {beta}-arrestin and the BBSome remains elusive. Here we find that, upon activation, ciliary GPCRs become tagged with K63-linked ubiquitin (K63Ub) chains in a {beta}-arrestin-dependent manner prior to BBSome-mediated exit. Removal of ubiquitin acceptor residues from the somatostatin receptor 3 (SSTR3) and from the orphan GPCR GPR161 demonstrates that ubiquitination of ciliary GPCRs is required for their regulated exit from cilia. Furthermore, targeting a K63Ub-specific deubiquitinase to cilia blocks the exit of GPR161, SSTR3 and Smoothened (SMO) from cilia. Finally, ubiquitinated proteins accumulate in cilia of mammalian photoreceptors and Chlamydomonas cells when BBSome function is compromised. We conclude that K63Ub chains mark GPCRs and other unwanted ciliary proteins for recognition by the ciliary exit machinery.
]]></description>
<dc:creator>Shinde, S. R.</dc:creator>
<dc:creator>Nager, A. R.</dc:creator>
<dc:creator>Nachury, M. V.</dc:creator>
<dc:date>2020-03-05</dc:date>
<dc:identifier>doi:10.1101/2020.03.04.977090</dc:identifier>
<dc:title><![CDATA[Lysine63-linked ubiquitin chains earmark GPCRs for BBSome-mediated removal from cilia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.05.979799v1?rss=1">
<title>
<![CDATA[
Hedgehog produced by the Drosophila wing imaginal disc induces distinct expression responses in three target tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.05.979799v1?rss=1</link>
<description><![CDATA[
Hedgehog (Hh) is an evolutionarily conserved signaling protein that has essential roles in animal development and homeostasis. We investigated Hh signaling in the region of the Drosophila wing imaginal disc that produces Hh and is near the tracheal air sac primordium (ASP) and myoblasts. Hh distributes in concentration gradients in the wing disc anterior compartment, ASP, and myoblasts and activates different sets of genes in each tissue. Some transcriptional targets of Hh signal transduction are common to the disc, ASP, and myoblasts, whereas others are tissue-specific. Signaling in the three tissues is cytoneme-mediated and cytoneme-dependent. We conclude that a single source of Hh in the wing disc regulates cell type-specific responses in three discreet target tissues.

SummaryHedgehog produced by the wing imaginal disc signals to wing disc, myoblast and tracheal cells
]]></description>
<dc:creator>Hatori, R.</dc:creator>
<dc:creator>Kornberg, T. B.</dc:creator>
<dc:date>2020-03-06</dc:date>
<dc:identifier>doi:10.1101/2020.03.05.979799</dc:identifier>
<dc:title><![CDATA[Hedgehog produced by the Drosophila wing imaginal disc induces distinct expression responses in three target tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.06.979583v1?rss=1">
<title>
<![CDATA[
Cytoskeletal tension forces mitohormesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.06.979583v1?rss=1</link>
<description><![CDATA[
Mitochondria control eukaryotic cell fate by producing the energy needed to support life and the signals required to execute programmed cell death. The biochemical milieu is known to affect mitochondrial function and contribute to the dysfunctional mitochondrial phenotypes implicated in cancer and the morbidities of ageing. However, the physical characteristics of the extracellular matrix are also altered in cancer and in aging tissues. We demonstrate that cells sense the physical properties of the extracellular matrix and activate a mitochondrial stress response that adaptively tunes mitochondrial function via SLC9A1-dependent ion exchange and HSF1-dependent transcription. Overall, our data indicate that adhesion-mediated mechanosignaling may play an unappreciated role in the altered mitochondrial functions observed in aging and cancer.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=154 SRC="FIGDIR/small/979583v3_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@1ecdaceorg.highwire.dtl.DTLVardef@1112c32org.highwire.dtl.DTLVardef@6c99a1org.highwire.dtl.DTLVardef@1da1e5f_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Tharp, K. M.</dc:creator>
<dc:creator>Higuchi-Sanabria, R.</dc:creator>
<dc:creator>Timblin, G.</dc:creator>
<dc:creator>Garzon-Coral, C.</dc:creator>
<dc:creator>Ford, B.</dc:creator>
<dc:creator>Schneider, C.</dc:creator>
<dc:creator>Muncie, J. M.</dc:creator>
<dc:creator>Stashko, C.</dc:creator>
<dc:creator>Daniele, J.</dc:creator>
<dc:creator>Frankino, P.</dc:creator>
<dc:creator>Manoli, S.</dc:creator>
<dc:creator>Shao, H.</dc:creator>
<dc:creator>Gestwicki, J.</dc:creator>
<dc:creator>Hellerstein, M.</dc:creator>
<dc:creator>Nomura, D. K.</dc:creator>
<dc:creator>Saijo, K.</dc:creator>
<dc:creator>Dunn, A.</dc:creator>
<dc:creator>Dillin, A.</dc:creator>
<dc:creator>Weaver, V. M.</dc:creator>
<dc:date>2020-03-07</dc:date>
<dc:identifier>doi:10.1101/2020.03.06.979583</dc:identifier>
<dc:title><![CDATA[Cytoskeletal tension forces mitohormesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.09.984773v1?rss=1">
<title>
<![CDATA[
An emergent population code in primary auditory cortex supports selective attention to spectral and temporal sound features 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.09.984773v1?rss=1</link>
<description><![CDATA[
Textbook descriptions of primary sensory cortex (PSC) revolve around single neurons representation of low-dimensional sensory features, such as visual object orientation in V1, location of somatic touch in S1, and sound frequency in A1. Typically, studies of PSC measure neurons responses along few (1 or 2) stimulus and/or behavioral dimensions. However, real-world stimuli usually vary along many feature dimensions and behavioral demands change constantly. In order to illuminate how A1 supports flexible perception in rich acoustic environments, we recorded from A1 neurons while rhesus macaques performed a feature-selective attention task. We presented sounds that varied along spectral and temporal feature dimensions (carrier bandwidth and temporal envelope, respectively). Within a block, subjects attended to one feature of the sound in a selective change detection task. We find that single neurons tend to be high-dimensional, in that they exhibit substantial mixed selectivity for both sound features, as well as task context. Contrary to common findings in many previous experiments, attention does not enhance the single-neuron representation of attended features in our data. However, a population-level analysis reveals that ensembles of neurons exhibit enhanced encoding of attended sound features, and this population code tracks subjects performance. Importantly, surrogate neural populations with intact single-neuron tuning but shuffled higher-order correlations among neurons failed to yield attention-related effects observed in the intact data. These results suggest that an emergent population code not measurable at the single-neuron level might constitute the functional unit of sensory representation in PSC.

SIGNIFICANCE STATEMENTThe ability to adapt to a dynamic sensory environment promotes a range of important natural behaviors. We recorded from single neurons in monkey primary auditory cortex while subjects attended to either the spectral or temporal features of complex sounds. Surprisingly, we find no average increase in responsiveness to, or encoding of, the attended feature across single neurons. However, when we pool the activity of the sampled neurons via targeted dimensionality reduction, we find enhanced population-level representation of the attended feature and suppression of the distractor feature. This dissociation of the effects of attention at the level of single neurons vs. the population highlights the synergistic nature of cortical sound encoding and enriches our understanding of sensory cortical function.
]]></description>
<dc:creator>Downer, J. D.</dc:creator>
<dc:creator>Verhein, J. R.</dc:creator>
<dc:creator>Rapone, B. C.</dc:creator>
<dc:creator>O'Connor, K. N.</dc:creator>
<dc:creator>Sutter, M. L.</dc:creator>
<dc:date>2020-03-10</dc:date>
<dc:identifier>doi:10.1101/2020.03.09.984773</dc:identifier>
<dc:title><![CDATA[An emergent population code in primary auditory cortex supports selective attention to spectral and temporal sound features]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.10.985796v1?rss=1">
<title>
<![CDATA[
ZENITH: A flow cytometry based method for functional profiling energy metabolism with single cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.10.985796v1?rss=1</link>
<description><![CDATA[
Energetic metabolism reprogramming is critical for cancer and immune responses. Current methods to functionally profile the global metabolic capacities and dependencies of cells are performed in bulk. We designed a simple method for complex metabolic profiling called SCENITH, for Single Cell ENergetIc metabolism by profilIng Translation inHibition. SCENITH allows for the study of metabolic responses in multiple cell types in parallel by flow cytometry. SCENITH is designed to perform metabolic studies ex vivo, particularly for rare cells in whole blood samples, avoiding metabolic biases introduced by culture media. We analyzed myeloid cells in solid tumors from patients and identified variable metabolic profiles, in ways that are not linked to their lineage nor their activation phenotype. SCENITH ability to reveal global metabolic functions and determine complex and linked immune-phenotypes in rare cell subpopulations will contribute to the information needed for evaluating therapeutic responses or patient stratification.
]]></description>
<dc:creator>ARGUELLO, R. J.</dc:creator>
<dc:creator>COMBES, A. J.</dc:creator>
<dc:creator>CHAR, R.</dc:creator>
<dc:creator>BOUSIQUOT, E.</dc:creator>
<dc:creator>GIGAN, J. P.</dc:creator>
<dc:creator>CAMOSSETO, V.</dc:creator>
<dc:creator>SAMAD, B.</dc:creator>
<dc:creator>TSUI, J.</dc:creator>
<dc:creator>YAN, P.</dc:creator>
<dc:creator>BOISSONNEAU, S.</dc:creator>
<dc:creator>FIGARELLA-BRANGER, D.</dc:creator>
<dc:creator>TABOURET, E.</dc:creator>
<dc:creator>GATTI, E.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>PIERRE, P.</dc:creator>
<dc:date>2020-03-11</dc:date>
<dc:identifier>doi:10.1101/2020.03.10.985796</dc:identifier>
<dc:title><![CDATA[ZENITH: A flow cytometry based method for functional profiling energy metabolism with single cell resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.10.986349v1?rss=1">
<title>
<![CDATA[
Hierarchical effects of choice related activity and neural encoding during feature selective attention 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.10.986349v1?rss=1</link>
<description><![CDATA[
Selective attention is necessary to sift through, form a coherent percept of, and make behavioral decisions on the vast amount of information present in most sensory environments. How and where selective attention is employed in cortex and how this perceptual information then informs the relevant behavioral decisions is still not well understood. Studies probing selective attention and decision making in visual cortex have been enlightening as to how sensory attention might work in that modality; whether or not similar mechanisms are employed in auditory attention is not yet clear. Therefore, we trained rhesus macaques on a feature selective attention task, where they switched between reporting changes in temporal (amplitude modulation, AM) and spectral (carrier bandwidth) features of a broadband noise stimulus. We investigated how the encoding of these features by single neurons in primary (A1) and secondary (lateral belt, ML) auditory cortex were affected by the different attention conditions. We found that neurons in A1 and ML showed mixed-selectivity to the sound and task features. We found no difference in AM encoding between the attention conditions. We found that choice-related activity in both A1 and ML neurons shifts between attentional conditions. This finding suggests that choice-related activity in auditory cortex does not simply reflect motor preparation or action, and supports the relationship between reported choice-related activity and the decision and perceptual process.

New & NoteworthyWe recorded from primary and secondary auditory cortex while monkeys performed a non-spatial feature attention task. Both areas exhibited rate-based choice-related activity. The manifestation of choice-related activity was attention-dependent, suggesting that choice-related activity in auditory cortex does not simply reflect arousal or motor influences, but relates to the specific perceptual choice. The lack of temporal-based choice activity is consistent with growing evidence that subcortical, but not cortical, single neurons inform decisions through temporal envelope following.
]]></description>
<dc:creator>Mohn, J. L.</dc:creator>
<dc:creator>Downer, J.</dc:creator>
<dc:creator>O'Connor, K. N.</dc:creator>
<dc:creator>Johnson, J. S.</dc:creator>
<dc:creator>Sutter, M. L.</dc:creator>
<dc:date>2020-03-11</dc:date>
<dc:identifier>doi:10.1101/2020.03.10.986349</dc:identifier>
<dc:title><![CDATA[Hierarchical effects of choice related activity and neural encoding during feature selective attention]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.12.989871v1?rss=1">
<title>
<![CDATA[
Scalable method for micro-CT analysis enables large scale quantitative characterization of brain lesions and implants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.12.989871v1?rss=1</link>
<description><![CDATA[
Anatomic evaluation is an important aspect of many studies in neuroscience; however, it often lacks information about the three-dimensional structure of the brain. Micro-CT imaging provides an excellent, nondestructive, method for the evaluation of brain structure, but current applications to neurophysiological or lesions studies require removal of the skull and hazardous chemicals, dehydration or embedding, limiting their scalability and utility. Here we present a protocol using eosin in combination with bone decalcification to enhance contrast in the tissue and then employ monochromatic and propagation phase-contrast micro-CT imaging to enable the imaging of brain structure with the preservation of the surrounding skull. Instead of relying on descriptive, time-consuming, or subjective methods, we develop simple quantitative analyses to map the locations of recording electrodes and to characterize the presence and extent of brain lesions.
]]></description>
<dc:creator>Kastner, D. B.</dc:creator>
<dc:creator>Kharazia, V.</dc:creator>
<dc:creator>Nevers, R.</dc:creator>
<dc:creator>Astudillo Maya, D.</dc:creator>
<dc:creator>Williams, G. M.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Smyth, C.</dc:creator>
<dc:creator>Della Santina, L.</dc:creator>
<dc:creator>Parkinson, D. Y.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:date>2020-03-14</dc:date>
<dc:identifier>doi:10.1101/2020.03.12.989871</dc:identifier>
<dc:title><![CDATA[Scalable method for micro-CT analysis enables large scale quantitative characterization of brain lesions and implants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.14.992073v1?rss=1">
<title>
<![CDATA[
Histone dynamics within the nucleosome octamer play a critical role in SNF2h-mediated nucleosome sliding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.14.992073v1?rss=1</link>
<description><![CDATA[
Elucidating the mechanisms by which ATP-dependent chromatin remodeling enzymes disrupt nucleosome structure is essential to understanding how chromatin states are established and maintained. A key finding informing remodeler mechanism is the observation that the dynamics of protein residues buried within the histone core of the nucleosome are used by specific remodelers to mobilize the nucleosome1. Recently, a study obtaining cryo-electron microscopy (cryo-EM) structures of ISWI-family remodelernucleosome complexes failed to observe stable conformational rearrangements in the histone octamer2. The authors of this study also failed to replicate the earlier finding that site-specifically restraining histone dynamics inhibits nucleosome sliding by ISWI-family remodelers1,2. In contrast, a recent cryo-EM structure detected asymmetric histone dynamics in an ISWI-nucleosome complex3. Here, using two different protocols, we replicate the original finding in Sinha et al.1 that dynamics within the histone core are important for nucleosome sliding by the human ISWI remodeler, SNF2h. These results firmly establish histone dynamics as an essential feature of ISWI-mediated nucleosome sliding and highlight the care required in designing and performing biochemical experiments investigating nucleosome dynamics using disulfide linkages.
]]></description>
<dc:creator>Gamarra, N.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:date>2020-03-16</dc:date>
<dc:identifier>doi:10.1101/2020.03.14.992073</dc:identifier>
<dc:title><![CDATA[Histone dynamics within the nucleosome octamer play a critical role in SNF2h-mediated nucleosome sliding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.19.999235v1?rss=1">
<title>
<![CDATA[
Evolution of an enzyme conformational ensemble guides design of an efficient biocatalyst 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.19.999235v1?rss=1</link>
<description><![CDATA[
The creation of artificial enzymes is a key objective of computational protein design. Although de novo enzymes have been successfully designed, these exhibit low catalytic efficiencies, requiring directed evolution to improve activity. Here, we used room-temperature X-ray crystallography to study changes in the conformational ensemble during evolution of the designed Kemp eliminase HG3 (kcat/KM 160 M-1s-1). We observed that catalytic residues were increasingly rigidified, the active site became better pre-organized, and its entrance was widened. Based on these observations, we engineered HG4, an efficient biocatalyst (kcat/KM 120,000 M-1s-1) containing active-site mutations found during evolution but not distal ones. HG4 structures revealed that its active site was pre-organized and rigidified for efficient catalysis. Our results show how directed evolution circumvents challenges inherent to enzyme design by shifting conformational ensembles to favor catalytically-productive sub-states, and suggest improvements to the design methodology that incorporate ensemble modeling of crystallographic data.
]]></description>
<dc:creator>Broom, A.</dc:creator>
<dc:creator>Rakotoharisoa, R. V.</dc:creator>
<dc:creator>Thompson, M. C.</dc:creator>
<dc:creator>Zarifi, N.</dc:creator>
<dc:creator>Nguyen, E.</dc:creator>
<dc:creator>Mukhametzhanov, N.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Chica, R. A.</dc:creator>
<dc:date>2020-03-20</dc:date>
<dc:identifier>doi:10.1101/2020.03.19.999235</dc:identifier>
<dc:title><![CDATA[Evolution of an enzyme conformational ensemble guides design of an efficient biocatalyst]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.20.000893v1?rss=1">
<title>
<![CDATA[
K2P channel C-type gating involves asymmetric selectivity filter order-disorder transitions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.20.000893v1?rss=1</link>
<description><![CDATA[
K2P channels regulate nervous, cardiovascular, and immune system functions1,2 through the action of their selectivity filter (C-type) gate3-6. Although structural studies show K2P conformations that impact activity7-13, no selectivity filter conformational changes have been observed. Here, combining K2P2.1 (TREK-1) X-ray crystallography in different potassium concentrations, potassium anomalous scattering, molecular dynamics, and functional studies, we uncover the unprecedented, asymmetric, potassium-dependent conformational changes underlying K2P C-type gating. Low potassium concentrations evoke conformational changes in selectivity filter strand 1 (SF1), selectivity filter strand 2 (SF2), and the SF2-transmembrane helix 4 loop (SF2-M4 loop) that destroy the S1 and S2 ion binding sites and are suppressed by C-type gate activator ML335. Shortening the uniquely long SF2-M4 loop to match the canonical length found in other potassium channels or disrupting the conserved Glu234 hydrogen bond network supporting this loop blunts C-type gate response to various physical and chemical stimuli. Glu234 network destabilization also compromises ion selectivity, but can be reversed by channel activation, indicating that the ion binding site loss reduces selectivity similar to other channels14. Together, our data establish that C-type gating occurs through potassium-dependent order-disorder transitions in the selectivity filter and adjacent loops that respond to gating cues relayed through the SF2-M4 loop. These findings underscore the potential for targeting the SF2-M4 loop for the development of new, selective K2P channel modulators.
]]></description>
<dc:creator>Lolicato, M.</dc:creator>
<dc:creator>Natale, A.</dc:creator>
<dc:creator>Aberemane-Ali, F.</dc:creator>
<dc:creator>Crottes, D.</dc:creator>
<dc:creator>Capponi, S.</dc:creator>
<dc:creator>Duman, R.</dc:creator>
<dc:creator>Wagner, A.</dc:creator>
<dc:creator>Rosenberg, J. M.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>Minor, D. L.</dc:creator>
<dc:date>2020-03-21</dc:date>
<dc:identifier>doi:10.1101/2020.03.20.000893</dc:identifier>
<dc:title><![CDATA[K2P channel C-type gating involves asymmetric selectivity filter order-disorder transitions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.22.002386v1?rss=1">
<title>
<![CDATA[
A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug Targets and Potential Drug-Repurposing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.22.002386v1?rss=1</link>
<description><![CDATA[
An outbreak of the novel coronavirus SARS-CoV-2, the causative agent of COVID-19 respiratory disease, has infected over 290,000 people since the end of 2019, killed over 12,000, and caused worldwide social and economic disruption1,2. There are currently no antiviral drugs with proven efficacy nor are there vaccines for its prevention. Unfortunately, the scientific community has little knowledge of the molecular details of SARS-CoV-2 infection. To illuminate this, we cloned, tagged and expressed 26 of the 29 viral proteins in human cells and identified the human proteins physically associated with each using affinity-purification mass spectrometry (AP-MS), which identified 332 high confidence SARS-CoV-2-human protein-protein interactions (PPIs). Among these, we identify 66 druggable human proteins or host factors targeted by 69 existing FDA-approved drugs, drugs in clinical trials and/or preclinical compounds, that we are currently evaluating for efficacy in live SARS-CoV-2 infection assays. The identification of host dependency factors mediating virus infection may provide key insights into effective molecular targets for developing broadly acting antiviral therapeutics against SARS-CoV-2 and other deadly coronavirus strains.
]]></description>
<dc:creator>Gordon, D. E.</dc:creator>
<dc:creator>Jang, G. M.</dc:creator>
<dc:creator>Bouhaddou, M.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Obernier, K.</dc:creator>
<dc:creator>O'Meara, M. J.</dc:creator>
<dc:creator>Guo, J. Z.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Tummino, T. A.</dc:creator>
<dc:creator>Huttenhain, R.</dc:creator>
<dc:creator>Kaake, R.</dc:creator>
<dc:creator>Richards, A. L.</dc:creator>
<dc:creator>Tutuncuoglu, B.</dc:creator>
<dc:creator>Foussard, H.</dc:creator>
<dc:creator>Batra, J.</dc:creator>
<dc:creator>Haas, K.</dc:creator>
<dc:creator>Modak, M.</dc:creator>
<dc:creator>Kim, M.</dc:creator>
<dc:creator>Haas, P.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Braberg, H.</dc:creator>
<dc:creator>Fabius, J. M.</dc:creator>
<dc:creator>Eckhardt, M.</dc:creator>
<dc:creator>Soucheray, M.</dc:creator>
<dc:creator>Brewer, M.</dc:creator>
<dc:creator>Cakir, M.</dc:creator>
<dc:creator>McGregor, M. J.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Naing, Z. Z. C.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Peng, S.</dc:creator>
<dc:creator>Kirby, I. T.</dc:creator>
<dc:creator>Melnyk, J. E.</dc:creator>
<dc:creator>Chorba, J. S.</dc:creator>
<dc:creator>Lou, K.</dc:creator>
<dc:creator>Dai, S. A.</dc:creator>
<dc:creator>Shen, W.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Barrio-Hernandez, I.</dc:creator>
<dc:creator>Memon, D.</dc:creator>
<dc:creator>Hernandez-Armenta, C.</dc:creator>
<dc:creator>Mathy, C. J. P.</dc:creator>
<dc:creator>Perica, T.</dc:creator>
<dc:creator>Pilla, K. B.</dc:creator>
<dc:date>2020-03-22</dc:date>
<dc:identifier>doi:10.1101/2020.03.22.002386</dc:identifier>
<dc:title><![CDATA[A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug Targets and Potential Drug-Repurposing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.30.016329v1?rss=1">
<title>
<![CDATA[
Coordinate genomic association of transcription factors controlled by an imported quorum sensing peptide in Cryptococcus neoformans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.30.016329v1?rss=1</link>
<description><![CDATA[
Qsp1 is a secreted quorum sensing peptide required for virulence of the fungal meningitis pathogen Cryptococcus neoformans. Qsp1 functions to control cell wall integrity in vegetatively growing cells and also functions in mating. Rather than acting on a cell surface receptor, Qsp1 is imported to act intracellularly via the predicted oligopeptide transporter Opt1. Here, we identify a transcription factor network as a target of Qsp1. Using whole-genome chromatin immunoprecipitation, we find Qsp1 controls the genomic associations of three transcription factors to genes whose outputs are regulated by Qsp1. One of these transcription factors, Cqs2, is also required for the action of Qsp1 during mating, indicating that it might be a shared proximal target of Qsp1. Consistent with this hypothesis, deletion of CQS2 impacts the binding of other network transcription factors specifically to Qsp1-regulated genes. These genetic and genomic studies illuminate mechanisms by which an imported peptide acts to modulate eukaryotic gene expression.

AUTHOR SUMMARYFor many fungal pathogens, the ability to adapt to changing and diverse environments forms the basis for their ability to infect and survive inside macrophages and other niches in the human body, and these changes are accomplished by transcription factors. Many pathogenic microbes coordinate their gene expression as a function of cell density in a process known as quorum sensing. Here, in the human fungal meningitis pathogen Cryptococcus neoformans, we find that an imported eukaryotic quorum sensing peptide that is important for virulence, Qsp1, controls the binding of three different transcription factors to promoters, thereby modulating the expression of Qsp1-regulated genes. This discovery reveals the mechanism for how an imported peptide affects gene expression.
]]></description>
<dc:creator>Summers, D. K.</dc:creator>
<dc:creator>Perry, D. S.</dc:creator>
<dc:creator>Rao, B.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:date>2020-03-30</dc:date>
<dc:identifier>doi:10.1101/2020.03.30.016329</dc:identifier>
<dc:title><![CDATA[Coordinate genomic association of transcription factors controlled by an imported quorum sensing peptide in Cryptococcus neoformans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.31.017434v1?rss=1">
<title>
<![CDATA[
A negative feedback loop mediated by the NR4A family of nuclear hormone receptors restrains expansion of B cells that receive signal one in the absence of signal two 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.31.017434v1?rss=1</link>
<description><![CDATA[
Ag stimulation (signal 1) triggers B cell activation and proliferation, and primes B cells to recruit, engage, and respond to T cell help (signal 2). However, failure to receive signal 2 within a defined window of time results in an abortive round of proliferation, followed by anergy or apoptosis. Although the molecular basis of T cell help has been extensively dissected, the mechanisms that restrain Ag-stimulated B cells, and enforce dependence upon co-stimulation, are incompletely understood. Nr4a1-3 encode a small family of orphan nuclear receptors that are rapidly induced by B cell receptor (BCR) stimulation, yet little is known about their function in humoral immune responses. Here we use germline and conditional loss-of-function mouse models to show that Nr4a1 and Nr4a3 play partially redundant roles to restrain both the survival and proliferation of B cells that receive signal 1 in the absence of co-stimulatory signals, and do so in part by repressing expression of BATF and consequently c-MYC. Correspondingly, Ab responses to TI-2 immunogens are enhanced in the absence of Nr4a1, but are unaltered in response to immunogens that incorporate co-stimulatory signals. Unexpectedly, we also identify a role for the NR4A family in restraining B cell access to T cell help by repressing expression of the T cell chemokines CCL3/4, as well as CD86 and ICAM1, and show that this is relevant under conditions of competition for limiting T cell help. Our studies collectively reveal a novel negative feedback loop mediated by the NR4A family that increases B cell dependence upon T cell help and restrains strongly Ag-activated B cell clones from monopolizing limiting amounts of T cell help. We speculate that this imposes B cell tolerance and dampens immunodominance to facilitate preservation of clonal diversity during an immune response.
]]></description>
<dc:creator>Tan, C.</dc:creator>
<dc:creator>Hiwa, R.</dc:creator>
<dc:creator>Mueller, J. L.</dc:creator>
<dc:creator>Vykunta, V.</dc:creator>
<dc:creator>Hibiya, K.</dc:creator>
<dc:creator>Noviski, M.</dc:creator>
<dc:creator>Huizar, J.</dc:creator>
<dc:creator>Brooks, J. F.</dc:creator>
<dc:creator>Garcia, J.</dc:creator>
<dc:creator>Heyn, C.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Zikherman, J.</dc:creator>
<dc:date>2020-04-01</dc:date>
<dc:identifier>doi:10.1101/2020.03.31.017434</dc:identifier>
<dc:title><![CDATA[A negative feedback loop mediated by the NR4A family of nuclear hormone receptors restrains expansion of B cells that receive signal one in the absence of signal two]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.12.038356v1?rss=1">
<title>
<![CDATA[
Lateralized role of prefrontal cortex in guiding orienting behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.12.038356v1?rss=1</link>
<description><![CDATA[
Orienting movements are essential to sensory-guided reward-seeking behaviors. Prefrontal cortex (PFC) is believed to exert top-down control over a range of goal-directed behaviors and is hypothesized to bias sensory-guided movements. However, the nature of PFC involvement in controlling sensory-guided orienting behaviors has remained largely unknown. Here, we trained rats on a delayed two-alternative forced-choice task requiring them to hold an orienting decision in working memory before execution is cued. Medial PFC (mPFC) Inactivation using either Muscimol or optogenetics impaired choice behavior. However, optogenetic impairment depended on the specific trial epoch during which inactivation took place. In particular, we found a lateralized role for mPFC during the presentation of instruction cues but this role became bilateral when inactivation occurred later in the delay period. Electrophysiological recording of multiple single-unit activity further provided evidence that this lateralized selectivity is cell-type specific. Our results suggest a previously unknown role of mPFC in mediating sensory-guided representation of orienting behavior and a potentially distinct cell-type specific role in shaping such representation across time.
]]></description>
<dc:creator>Mohebi, A.</dc:creator>
<dc:creator>Oweiss, K. G.</dc:creator>
<dc:date>2020-04-13</dc:date>
<dc:identifier>doi:10.1101/2020.04.12.038356</dc:identifier>
<dc:title><![CDATA[Lateralized role of prefrontal cortex in guiding orienting behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.13.039297v1?rss=1">
<title>
<![CDATA[
Rapid, precise quantification of large DNA excisions and inversions by ddPCR 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.13.039297v1?rss=1</link>
<description><![CDATA[
The excision of genomic sequences using paired CRISPR-Cas nucleases is a powerful tool to study gene function, create disease models and holds promise for therapeutic gene editing. However, our understanding of the factors that favor efficient excision is limited by the lack of a rapid, accurate measurement of DNA excision outcomes that is free of amplification bias. Here, we introduce ddXR (droplet digital PCR eXcision Reporter), a method that enables the accurate and sensitive detection of excisions and inversions independent of length. The method can be completed in a few hours without the need for next-generation sequencing. The ddXR method uncovered unexpectedly high rates of large (>20 kb) excisions and inversions, while also revealing a surprisingly low dependence on linear distance, up to 170 kb. We further modified the method to measure precise repair of excision junctions and allele-specific excision, with important implications for disease modeling and therapeutic gene editing.
]]></description>
<dc:creator>Watry, H. L.</dc:creator>
<dc:creator>Feliciano, C. M.</dc:creator>
<dc:creator>Gjoni, K.</dc:creator>
<dc:creator>Takahashi, G.</dc:creator>
<dc:creator>Miyaoka, Y.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:creator>Judge, L. M.</dc:creator>
<dc:date>2020-04-14</dc:date>
<dc:identifier>doi:10.1101/2020.04.13.039297</dc:identifier>
<dc:title><![CDATA[Rapid, precise quantification of large DNA excisions and inversions by ddPCR]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.13.039677v1?rss=1">
<title>
<![CDATA[
Small molecule cognitive enhancer reverses age-related memory decline in mice. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.13.039677v1?rss=1</link>
<description><![CDATA[
With increased life expectancy age-associated cognitive decline becomes a growing concern, even in the absence of recognizable neurodegenerative disease. The integrated stress response (ISR) is activated during aging and contributes to age-related brain phenotypes. We demonstrate that treatment with the drug-like small-molecule ISR inhibitor ISRIB reverses ISR activation in the brain, as indicated by decreased levels of activating transcription factor 4 (ATF4) and phosphorylated eukaryotic translation initiation factor eIF2. Furthermore, ISRIB treatment reverses spatial memory deficits and ameliorates working memory in old mice. At the cellular level in the hippocampus, ISR inhibition i) rescues intrinsic neuronal electrophysiological properties, ii) restores spine density and iii) reduces immune profiles, specifically interferon and T cell-mediated responses. Thus, pharmacological interference with the ISR emerges as a promising intervention strategy for combating age-related cognitive decline in otherwise healthy individuals.

ONE SENTENCE SUMMARYInhibition of the integrated stress response restores neuronal and immune dysfunction and alleviates memory deficits in aged mice.
]]></description>
<dc:creator>Krukowski, K.</dc:creator>
<dc:creator>Nolan, A.</dc:creator>
<dc:creator>Frias, E. S.</dc:creator>
<dc:creator>Boone, M.</dc:creator>
<dc:creator>Ureta, G.</dc:creator>
<dc:creator>Grue, K.</dc:creator>
<dc:creator>Paladini, M.-S.</dc:creator>
<dc:creator>Elizarraras, E.</dc:creator>
<dc:creator>Delgado, L.</dc:creator>
<dc:creator>Bernales, S.</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:creator>Rosi, S.</dc:creator>
<dc:date>2020-04-15</dc:date>
<dc:identifier>doi:10.1101/2020.04.13.039677</dc:identifier>
<dc:title><![CDATA[Small molecule cognitive enhancer reverses age-related memory decline in mice.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.15.037325v1?rss=1">
<title>
<![CDATA[
Blood RNA Profiles are Diagnostic for Severity in Human Acute Spinal Cord Injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.15.037325v1?rss=1</link>
<description><![CDATA[
Biomarkers of spinal cord injury (SCI) could help determine the severity of the injury and facilitate early critical care decision making. We analyzed global gene expression in peripheral white blood cells during the acute injury phase and identified 197 genes whose expression changed after SCI compared to healthy and trauma controls and in direct relation to SCI severity. Unsupervised co-expression network analysis identified several gene modules that predicted injury severity (AIS grades) with an overall accuracy of 72.7% and included signatures of immune cell subtypes. Our findings indicate that global transcriptomic changes in peripheral blood cells have diagnostic and potentially prognostic value for SCI severity.
]]></description>
<dc:creator>Kyritsis, N.</dc:creator>
<dc:creator>Torres-Espin, A.</dc:creator>
<dc:creator>Schupp, P. G.</dc:creator>
<dc:creator>Huie, J. R.</dc:creator>
<dc:creator>Chou, A.</dc:creator>
<dc:creator>Duong Fernandez, X.</dc:creator>
<dc:creator>Thomas, L. H.</dc:creator>
<dc:creator>Tsolinas, R. E.</dc:creator>
<dc:creator>Hemmerle, D.</dc:creator>
<dc:creator>Pascual, L. U.</dc:creator>
<dc:creator>Singh, V.</dc:creator>
<dc:creator>Pan, J. Z.</dc:creator>
<dc:creator>Talbott, J. F.</dc:creator>
<dc:creator>Whetstone, W. D.</dc:creator>
<dc:creator>Burke, J. F.</dc:creator>
<dc:creator>DiGiorgio, A. M.</dc:creator>
<dc:creator>Weinstein, P. R.</dc:creator>
<dc:creator>Manley, G. T.</dc:creator>
<dc:creator>Dhall, S. S.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Oldham, M. C.</dc:creator>
<dc:creator>Bresnahan, J. C.</dc:creator>
<dc:creator>Beattie, M. S.</dc:creator>
<dc:date>2020-04-17</dc:date>
<dc:identifier>doi:10.1101/2020.04.15.037325</dc:identifier>
<dc:title><![CDATA[Blood RNA Profiles are Diagnostic for Severity in Human Acute Spinal Cord Injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.16.045245v1?rss=1">
<title>
<![CDATA[
Continuous lineage recording reveals rapid, multidirectional metastasis in a lung cancer xenograft model in mouse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.16.045245v1?rss=1</link>
<description><![CDATA[
Cancer progression is characterized by rare, transient events which are nonetheless highly consequential to disease etiology and mortality. Detailed cell phylogenies can recount the history and chronology of these critical events - including metastatic seeding. Here, we applied our Cas9-based lineage tracer to study the subclonal dynamics of metastasis in a lung cancer xenograft mouse model, revealing the underlying rates, routes, and drivers of metastasis. We report deeply resolved phylogenies for tens of thousands of metastatically disseminated cancer cells. We observe surprisingly diverse metastatic phenotypes, ranging from metastasis-incompetent to aggressive populations. These phenotypic distinctions result from pre-existing, heritable, and characteristic differences in gene expression, and we demonstrate that these differentially expressed genes can drive invasiveness. Furthermore, metastases transit via diverse, multidirectional tissue routes and seeding topologies. Our work demonstrates the power of tracing cancer progression at unprecedented resolution and scale.

One Sentence SummarySingle-cell lineage tracing and RNA-seq capture diverse metastatic behaviors and drivers in lung cancer xenografts in mice.
]]></description>
<dc:creator>Quinn, J. J.</dc:creator>
<dc:creator>Jones, M. G.</dc:creator>
<dc:creator>Okimoto, R. A.</dc:creator>
<dc:creator>Yosef, N.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:date>2020-04-17</dc:date>
<dc:identifier>doi:10.1101/2020.04.16.045245</dc:identifier>
<dc:title><![CDATA[Continuous lineage recording reveals rapid, multidirectional metastasis in a lung cancer xenograft model in mouse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.18.043927v1?rss=1">
<title>
<![CDATA[
SMARCA4 supports the oncogenic landscape of KRAS-driven lung tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.18.043927v1?rss=1</link>
<description><![CDATA[
Cancer resequencing studies identify recurrent mutations in the switch/sucrose non-fermentable (SWI/SNF) complex at an unexpectedly high frequency across many cancer types. Some SWI/SNF mutations appear to be loss-of-function events, implying that the intact SWI/SNF complex is tumor suppressive. We examined the distribution and function of SMARCA4 mutations, the most frequently mutated SWI/SNF complex gene in lung adenocarcinoma, using human cancers, cell lines and mouse model systems. We found that lung adenocarcinomas harboring activated oncogenes have fewer deleterious mutations in SMARCA4 and express higher levels of the mRNA than cancers without activated oncogenes, indicating distinct dependencies on SMARCA4 in these two settings. Surprisingly, intact Smarca4 promoted the growth and tumorgenicity of KrasG12D-driven mouse lung tumors and human cells. Mechanistically, we found that Smarca4 supports the oncogenic transcriptional/signaling landscape of KrasG12D-driven mouse lung cancer. This dependency on the chromatin maintenance machinery in established cancer cells support treatments directed towards pathogenic SWI/SNF complexes in lung adenocarcinoma and other malignancies.
]]></description>
<dc:creator>Collisson, E.</dc:creator>
<dc:date>2020-04-20</dc:date>
<dc:identifier>doi:10.1101/2020.04.18.043927</dc:identifier>
<dc:title><![CDATA[SMARCA4 supports the oncogenic landscape of KRAS-driven lung tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.20.051888v1?rss=1">
<title>
<![CDATA[
Role of c-Met/β1 integrin complex in the metastatic cascade 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.20.051888v1?rss=1</link>
<description><![CDATA[
Metastases cause 90% of human cancer deaths. The metastatic cascade involves local invasion, intravasation, extravasation, metastatic site colonization, and proliferation. While individual mediators of these processes have been investigated, interactions between these mediators remain less well defined. We previously identified a structural complex between receptor tyrosine kinase c-Met and {beta}1 integrin in metastases. Using novel cell culture and in vivo assays, we found that c-Met/{beta}1 complex induction promotes breast cancer intravasation and adhesion to the vessel wall, but does not increase extravasation. These effects may be driven by the ability of the c-Met/{beta}1 complex to increase mesenchymal and stem cell characteristics. Multiplex transcriptomic analysis revealed upregulated Wnt and hedgehog pathways after c-Met/{beta}1 complex induction. A {beta}1 integrin point mutation that prevented binding to c-Met reduced intravasation. OS2966, a therapeutic antibody disrupting c-Met/{beta}1 binding, decreased invasion and mesenchymal gene expression and morphology of breast cancer cells. Bone-seeking breast cancer cells exhibited higher c-Met/{beta}1 complex levels than parental controls and preferentially adhere to tissue-specific matrix. Patient bone metastases demonstrated higher c-Met/{beta}1 levels than brain metastases. Thus, the c-Met/{beta}1 complex drives breast cancer cell intravasation and preferential affinity for bone tissue-specific matrix. Pharmacological targeting of the complex may prevent metastases, particularly osseous metastases.
]]></description>
<dc:creator>Lau, D.</dc:creator>
<dc:creator>Wadhwa, H.</dc:creator>
<dc:creator>Sudhir, S.</dc:creator>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Chandra, A.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Spatz, J.</dc:creator>
<dc:creator>Shah, S. S.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Safaee, M.</dc:creator>
<dc:creator>Yagnik, G.</dc:creator>
<dc:creator>Jahangiri, A.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:date>2020-04-22</dc:date>
<dc:identifier>doi:10.1101/2020.04.20.051888</dc:identifier>
<dc:title><![CDATA[Role of c-Met/β1 integrin complex in the metastatic cascade]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.23.057026v1?rss=1">
<title>
<![CDATA[
G protein-regulated endocytic trafficking of adenylyl cyclase type 9 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.23.057026v1?rss=1</link>
<description><![CDATA[
GPCRs are increasingly recognized to initiate signaling via heterotrimeric G proteins as they move through the endocytic network, but little is known about how relevant G protein effectors are localized. Here we report dynamic trafficking of adenylyl cyclase type 9 (AC9) from the plasma membrane to endosomes, while adenylyl cyclase type 1 (AC1) remains in the plasma membrane, and stimulation of AC9 trafficking by ligand-induced activation of Gs-coupled GPCRs or Gs. AC9 transits a similar dynamin-dependent early endocytic pathway as activated GPCRs but, in contrast to GPCR trafficking which is regulated by {beta}-arrestin but not Gs, AC9 trafficking is regulated by Gs but not {beta}-arrestin. We also show that AC9, but not AC1, contributes to cAMP production from endosomes. These results reveal dynamic and isoform-specific trafficking of adenylyl cyclase in the endocytic network, and a discrete role of a heterotrimeric G protein in controlling subcellular location of a relevant effector.
]]></description>
<dc:creator>Lazar, A. M.</dc:creator>
<dc:creator>Irannejad, R.</dc:creator>
<dc:creator>Baldwin, T. A.</dc:creator>
<dc:creator>Sundaram, A. B.</dc:creator>
<dc:creator>Gutkind, J. S.</dc:creator>
<dc:creator>Inoue, A.</dc:creator>
<dc:creator>Dessauer, C. W.</dc:creator>
<dc:creator>Von Zastrow, M.</dc:creator>
<dc:date>2020-04-25</dc:date>
<dc:identifier>doi:10.1101/2020.04.23.057026</dc:identifier>
<dc:title><![CDATA[G protein-regulated endocytic trafficking of adenylyl cyclase type 9]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.24.059840v1?rss=1">
<title>
<![CDATA[
A splicing isoform of GPR56 mediates microglial synaptic refinement via phosphatidylserine binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.24.059840v1?rss=1</link>
<description><![CDATA[
Developmental synaptic remodeling is important for the formation of precise neural circuitry and its disruption has been linked to neurodevelopmental disorders such as autism and schizophrenia. Microglia prune synapses, but integration of this synapse pruning with overlapping and concurrent neurodevelopmental processes remains elusive. Adhesion G protein-coupled receptor ADGRG1/GPR56 controls multiple aspects of brain development in a cell type-specific manner: in neural progenitor cells, GPR56 regulates cortical lamination, whereas in oligodendrocyte progenitor cells, GPR56 controls developmental myelination and myelin repair. Here, we show that microglial GPR56 maintains appropriate synaptic numbers in several brain regions in a time- and circuit-dependent fashion. Phosphatidylserine (PS) on pre-synaptic elements binds GPR56 in a domain-specific manner, and microglia-specific deletion of Gpr56 leads to increased synapses as a result of reduced microglial engulfment of PS+ pre-synaptic inputs. Remarkably, a particular alternatively spliced isoform of GPR56 is selectively required for microglia-mediated synaptic pruning. Our present data provide a ligand- and isoform-specific mechanism underlying microglial GPR56-mediated synapse pruning in the context of complex neurodevelopmental processes.
]]></description>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Chiou, B.</dc:creator>
<dc:creator>Gilman, C. K.</dc:creator>
<dc:creator>Luo, R.</dc:creator>
<dc:creator>Koshi, T.</dc:creator>
<dc:creator>Yu, D.</dc:creator>
<dc:creator>Oak, H. C.</dc:creator>
<dc:creator>Giera, S.</dc:creator>
<dc:creator>Johnson-Venkatesh, E.</dc:creator>
<dc:creator>Muthukumar, A. K.</dc:creator>
<dc:creator>Stevens, B.</dc:creator>
<dc:creator>Umemori, H.</dc:creator>
<dc:creator>Piao, X.</dc:creator>
<dc:date>2020-04-25</dc:date>
<dc:identifier>doi:10.1101/2020.04.24.059840</dc:identifier>
<dc:title><![CDATA[A splicing isoform of GPR56 mediates microglial synaptic refinement via phosphatidylserine binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.24.060558v1?rss=1">
<title>
<![CDATA[
Sex-Specific Cross Tissue Meta-Analysis Identifies Immune Dysregulation in Women with Alzheimer's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.24.060558v1?rss=1</link>
<description><![CDATA[
In spite of evidence of females having a greater lifetime risk of developing Alzheimers Disease (AD) and greater apolipoprotein E4-related (apoE4) AD risk compared to males, molecular signatures underlying these findings remain elusive. We took a meta-analysis approach to study gene expression in the brains of 1,084 AD patients and age-matched controls and whole blood from 645 AD patients and age-matched controls. Gene-expression, network-based analysis and cell type deconvolution approaches revealed a consistent immune signature in the brain and blood of female AD patients that was absent in males. Machine learning-based classification of AD using gene expression from whole blood in addition to clinical features revealed an improvement in classification accuracy upon stratifying by sex, achieving an AUROC of 0.91 for females and 0.80 for males. These results help identify sex and apoE4 genotype-specific transcriptomic signatures of AD and underscore the importance of considering sex in the development of biomarkers and therapeutic strategies for AD.
]]></description>
<dc:creator>Paranjpe, M. D.</dc:creator>
<dc:creator>Belonwu, S.</dc:creator>
<dc:creator>Wang, J. K.</dc:creator>
<dc:creator>Oskotsky, T.</dc:creator>
<dc:creator>Gupta, A.</dc:creator>
<dc:creator>Taubes, A.</dc:creator>
<dc:creator>Zalocusky, K.</dc:creator>
<dc:creator>Paranjpe, I.</dc:creator>
<dc:creator>Glicksberg, B. S.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2020-04-25</dc:date>
<dc:identifier>doi:10.1101/2020.04.24.060558</dc:identifier>
<dc:title><![CDATA[Sex-Specific Cross Tissue Meta-Analysis Identifies Immune Dysregulation in Women with Alzheimer's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.02.070680v1?rss=1">
<title>
<![CDATA[
The mitotic protein NuMA plays a spindle-independent role in nuclear formation and mechanics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.02.070680v1?rss=1</link>
<description><![CDATA[
Eukaryotic cells typically form a single, round nucleus after mitosis, and failures to do so can compromise genomic integrity. How mammalian cells form such a nucleus remains incompletely understood. NuMA is a spindle protein whose disruption results in nuclear fragmentation. What role NuMA plays in nuclear integrity, or whether its perceived role stems from its spindle function, is unclear. Here, we use live imaging to demonstrate that NuMA plays a spindle-independent role in forming a single, round nucleus. NuMA keeps the decondensing chromosome mass compact at mitotic exit, and promotes a mechanically robust nucleus. NuMAs C-terminus binds DNA in vitro and chromosomes in interphase, while its coiled-coil acts as a regulatory and structural hub: it prevents NuMA from binding chromosomes at mitosis, regulates its nuclear mobility and is essential for nuclear formation. Thus, NuMA plays a long-range structural role in building and maintaining an intact nucleus, as it does for the spindle, playing a protective role over the cell cycle.
]]></description>
<dc:creator>Serra-Marques, A.</dc:creator>
<dc:creator>Houtekamer, R.</dc:creator>
<dc:creator>Hintzen, D.</dc:creator>
<dc:creator>Canty, J. T.</dc:creator>
<dc:creator>Yildiz, A.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2020-05-03</dc:date>
<dc:identifier>doi:10.1101/2020.05.02.070680</dc:identifier>
<dc:title><![CDATA[The mitotic protein NuMA plays a spindle-independent role in nuclear formation and mechanics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.05.079418v1?rss=1">
<title>
<![CDATA[
Immunodietica: interrogating the role of diet in autoimmune disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.05.079418v1?rss=1</link>
<description><![CDATA[
Diet is an environmental factor in autoimmune disorders, where the immune system erroneously destroys ones own tissues. Yet, interactions between diet and autoimmunity remain largely unexplored, particularly the impact of immunogenetics, ones human leukocyte antigen (HLA) allele make-up, in this interplay. Here, we interrogated animals and plants for the presence of epitopes implicated in human autoimmune diseases. We mapped autoimmune epitope distribution across organisms and determined their tissue expression pattern. Interestingly, diet-derived epitopes implicated in a disease were more likely to bind to HLA alleles associated with that disease than to protective alleles, with visible differences between organisms with similar autoimmune epitope content. We then analyzed an individuals HLA haplotype, generating a personalized heatmap of potential dietary autoimmune triggers. Our work uncovered differences in autoimmunogenic potential across food sources and revealed differential binding of diet-derived epitopes to autoimmune disease-associated HLA alleles, shedding light on the impact of diet on autoimmunity.
]]></description>
<dc:creator>Gershteyn, I. M.</dc:creator>
<dc:creator>Burov, A. A.</dc:creator>
<dc:creator>Miao, B. Y.</dc:creator>
<dc:creator>Morais, V. H.</dc:creator>
<dc:creator>Ferreira, L. M. R.</dc:creator>
<dc:date>2020-05-07</dc:date>
<dc:identifier>doi:10.1101/2020.05.05.079418</dc:identifier>
<dc:title><![CDATA[Immunodietica: interrogating the role of diet in autoimmune disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.09.086264v1?rss=1">
<title>
<![CDATA[
Deep profiling of protease substrate specificity enabled by dual random and scanned human proteome substrate phage libraries 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.09.086264v1?rss=1</link>
<description><![CDATA[
Proteolysis is a major post-translational regulator of biology both inside and outside of cells. Broad identification of optimal cleavage sites and natural substrates of proteases is critical for drug discovery and to understand protease biology. Here we present a method that employs two genetically encoded substrate phage display libraries coupled with next generation sequencing (SPD-NGS) that allows up to 10,000-fold deeper sequence coverage of the typical 6 to 8 residue protease cleavage sites compared to state-of-the-art synthetic peptide libraries or proteomics. We applied SPD-NGS to two classes of proteases, the intracellular caspases 2, 3, 6, 7 and 8, and the ectodomains of the membrane sheddases, ADAMs 10 and 17. The first library (Lib 10AA) was used to determine substrate cleavage motifs. Lib 10AA contains a highly diverse randomized 10-mer substrate peptide sequences (109 unique members) that was displayed mono-valently on filamentous phage and bound to magnetic beads via an N-terminal biotin. The protease was allowed to cleave the SPD beads, and the released phage subjected to up to three total rounds of positive selection followed by next generation sequencing (NGS). This allowed us to identify from 104 to 105 unique cleavage sites over a 1000-fold dynamic range of NGS counts (ranging from 3-4000), and produced consensus and optimal cleavage motifs based positional sequencing scoring matrices that closely matched synthetic peptide data. A second SPD-NGS library (Lib hP) was constructed that allowed us to identify candidate human proteome sequences. Lib hP displayed virtually the entire human proteome tiled in contiguous 49AA sequences with 25AA overlaps (nearly 1 million members). After three rounds of positive selection we identified up to 104 natural linear cut sites depending on the protease and captured most of the examples previously identified by proteomics (ranging from 30 to 1500) and predicted 10 to 100-fold more. Structural bioinformatics was used to facilitate the identification of candidate natural protein substrates. SPD-NGS is rapid, reproducible, simple to perform and analyze, inexpensive, renewable, with unprecedented depth of coverage for substrate sequences. SPD-NGS is an important tool for protease biologists interested protease specificity for specific assays and inhibitors and to facilitate identification of natural protein substrates.
]]></description>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>ODonovan, B. K.</dc:creator>
<dc:creator>Zou, J.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2020-05-10</dc:date>
<dc:identifier>doi:10.1101/2020.05.09.086264</dc:identifier>
<dc:title><![CDATA[Deep profiling of protease substrate specificity enabled by dual random and scanned human proteome substrate phage libraries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.12.088971v1?rss=1">
<title>
<![CDATA[
Exploiting KRAS-driven Ferroaddiction in Cancer Through Ferrous Iron-Activatable Drug Conjugates (FeADC) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.12.088971v1?rss=1</link>
<description><![CDATA[
KRAS mutations cause a quarter of cancer mortality and most are undruggable. Several inhibitors of the MAPK pathway are FDA approved but poorly tolerated at dosages required to adequately extinguish RAS/RAF/MAPK signaling. We found that oncogenic KRAS signaling induces ferrous iron (Fe2+) accumulation early in and throughout KRAS-mediated transformation. We used an FDA-approved MEK inhibitor to produce a prototypical Ferrous Iron-Activatable Drug Conjugate (FeADC) which achieved potent MAPK blockade in tumor cells while sparing normal tissues. This innovation allowed sustainable, effective treatment of tumor bearing animals, with tumor-selective drug activation producing superior systemic tolerability. Ferrous iron accumulation is an exploitable feature of KRAS transformation and FeADCs hold promise for improving treatment of KRAS-driven solid tumors.
]]></description>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Muir, R. K.</dc:creator>
<dc:creator>Gonciarz, R. L.</dc:creator>
<dc:creator>Olshen, A. B.</dc:creator>
<dc:creator>Yeh, I.</dc:creator>
<dc:creator>Hann, B. C.</dc:creator>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Wang, Y.-h.</dc:creator>
<dc:creator>Korkola, J.</dc:creator>
<dc:creator>Behr, S. C.</dc:creator>
<dc:creator>Evans, M. J.</dc:creator>
<dc:creator>collisson, e.</dc:creator>
<dc:creator>Renslo, A.</dc:creator>
<dc:date>2020-05-14</dc:date>
<dc:identifier>doi:10.1101/2020.05.12.088971</dc:identifier>
<dc:title><![CDATA[Exploiting KRAS-driven Ferroaddiction in Cancer Through Ferrous Iron-Activatable Drug Conjugates (FeADC)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.12.091082v1?rss=1">
<title>
<![CDATA[
Androgen regulates SARS-CoV-2 receptor levels and is associated with severe COVID-19 symptoms in men 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.12.091082v1?rss=1</link>
<description><![CDATA[
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has led to a global health crisis, and yet our understanding of the disease pathophysiology and potential treatment options remains limited. SARS-CoV-2 infection occurs through binding and internalization of the viral spike protein to angiotensin converting enzyme 2 (ACE2) on the host cell membrane. Lethal complications are caused by damage and failure of vital organs that express high levels of ACE2, including the lungs, the heart and the kidneys. Here, we established a high-throughput drug screening strategy to identify therapeutic candidates that reduce ACE2 levels in human embryonic stem cell (hESC) derived cardiac cells. Drug target analysis of validated hit compounds, including 5 alpha reductase inhibitors, revealed androgen signaling as a key modulator of ACE2 levels. Treatment with the 5 alpha reductase inhibitor dutasteride reduced ACE2 levels and internalization of recombinant spike receptor binding domain (Spike-RBD) in hESC-derived cardiac cells and human alveolar epithelial cells. Finally, clinical data on coronavirus disease 2019 (COVID-19) patients demonstrated that abnormal androgen states are significantly associated with severe disease complications and cardiac injury as measured by blood troponin T levels. These findings provide important insights on the mechanism of increased disease susceptibility in male COVID-19 patients and identify androgen receptor inhibition as a potential therapeutic strategy.
]]></description>
<dc:creator>Ghazizadeh, Z.</dc:creator>
<dc:creator>Majd, H.</dc:creator>
<dc:creator>Richter, M.</dc:creator>
<dc:creator>Samuel, R.</dc:creator>
<dc:creator>Zekavat, S. M.</dc:creator>
<dc:creator>Asgharian, H.</dc:creator>
<dc:creator>Farahvashi, S.</dc:creator>
<dc:creator>Kalantari, A.</dc:creator>
<dc:creator>Ramirez, J.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Natarajan, P.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:date>2020-05-12</dc:date>
<dc:identifier>doi:10.1101/2020.05.12.091082</dc:identifier>
<dc:title><![CDATA[Androgen regulates SARS-CoV-2 receptor levels and is associated with severe COVID-19 symptoms in men]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.14.096560v1?rss=1">
<title>
<![CDATA[
A Molecular Mechanism for Probabilistic Bet-hedging and its Role in Viral Latency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.14.096560v1?rss=1</link>
<description><![CDATA[
Probabilistic bet hedging, a strategy to maximize fitness in unpredictable environments by matching phenotypic variability to environmental variability, is theorized to account for the evolution of various fate-specification decisions, including viral latency. However, the molecular mechanisms underlying bet-hedging remain unclear. Here, we report that large variability in protein abundance within individual herpesvirus virion particles enables probabilistic bet hedging between viral replication and latency. Super-resolution imaging of individual virions of the human herpesvirus cytomegalovirus (CMV) showed that virion-to-virion levels of pp71 tegument protein--the major viral transactivator protein--exhibit extreme variability. This super-Poissonian tegument variability promoted alternate replicative strategies: high virion pp71 levels enhance viral replicative fitness but, strikingly, impede silencing, whereas low virion pp71 levels reduce fitness but promote silencing. Overall, the results indicate that stochastic tegument packaging provides a mechanism enabling probabilistic bet hedging between viral replication and latency.

SIGNIFICANCEProbabilistic bet hedging is a generalized diversification strategy to maximize fitness in unpredictable environments, and has been proposed as an evolutionary basis for herpesvirus latency. However, the molecular mechanisms enabling probabilistic bet hedging have remained elusive. Here, we find that the human herpesvirus cytomegalovirus--a major cause of birth defects and transplant failures--utilizes stochastic variability in the abundance of a protein packaged into individual viral particles to enable probabilistic bet hedging between alternate viral states.
]]></description>
<dc:creator>Chaturvedi, S.</dc:creator>
<dc:creator>Klein, J.</dc:creator>
<dc:creator>Vardi, N.</dc:creator>
<dc:creator>Bolovan-Fritts, C.</dc:creator>
<dc:creator>Wolf, M.</dc:creator>
<dc:creator>Du, K.</dc:creator>
<dc:creator>Mlera, L.</dc:creator>
<dc:creator>Calvert, M.</dc:creator>
<dc:creator>Moorman, N. J.</dc:creator>
<dc:creator>Goodrum, F.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Weinberger, L. S.</dc:creator>
<dc:date>2020-05-15</dc:date>
<dc:identifier>doi:10.1101/2020.05.14.096560</dc:identifier>
<dc:title><![CDATA[A Molecular Mechanism for Probabilistic Bet-hedging and its Role in Viral Latency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.15.098012v1?rss=1">
<title>
<![CDATA[
Polarized endosome dynamics engage cytosolic Par-3 and dynein during asymmetric division 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.15.098012v1?rss=1</link>
<description><![CDATA[
Asymmetric cell division (ACD), which produces two daughters with different fates, is fundamental for generating cellular diversity. In the developing embryos of both invertebrates and vertebrates, asymmetrically dividing progenitors generate daughter cells with differential activity of Notch signaling1-7, a key regulator of cell fate decisions8,9. The cell polarity regulator Par-3 is critical for establishing this Notch asymmetry1,4,6, but the underlying mechanisms are not understood. Here, employing in vivo time-lapse imaging in the developing zebrafish forebrain during the mitotic cycle of radial glia, the principal vertebrate neural stem cells10,11, we show that during ACD, endosomes containing the Notch ligand Delta D (Dld) undergo convergent movement toward the cleavage plane, followed by preferential segregation into the posterior (and subsequently basal) Notchhi daughter. This asymmetric segregation requires the activity of Par-3 and the dynein motor complex. Employing label-retention expansion microscopy, we further detect Par-3 in the cytosol in association with the dynein light intermediate chain 1 (DLIC1) on Dld endosomes, suggesting a direct involvement of Par-3 in dynein-mediated polarized transport of Notch signaling endosomes. Our data reveal an unanticipated mechanism by which Par-3 regulates cell fate decision by directly polarizing Notch signaling components during ACD.
]]></description>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Tong, K.</dc:creator>
<dc:creator>Chen, x.</dc:creator>
<dc:creator>Yang, B.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Dai, Z.</dc:creator>
<dc:creator>Shi, X.</dc:creator>
<dc:creator>Seiple, I.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:date>2020-05-16</dc:date>
<dc:identifier>doi:10.1101/2020.05.15.098012</dc:identifier>
<dc:title><![CDATA[Polarized endosome dynamics engage cytosolic Par-3 and dynein during asymmetric division]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.15.098699v1?rss=1">
<title>
<![CDATA[
Hsp70 chaperone blocks α-synuclein oligomer formation via a novel engagement mechanism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.15.098699v1?rss=1</link>
<description><![CDATA[
Over-expression and aggregation of -synuclein (ASyn) are linked to the onset and pathology of Parkinsons disease and related synucleinopathies. Elevated levels of the stress induced chaperone, Hsp70, protects against ASyn misfolding and ASyn-driven neurodegeneration in cell and animal models, yet there is minimal mechanistic understanding of this important protective pathway. It is generally assumed that Hsp70 binds to ASyn using its canonical and promiscuous substrate-binding cleft to limit aggregation. Here we report that this activity is due to a novel and unexpected mode of Hsp70 action, involving neither ATP nor the typical substrate-binding cleft. We use novel ASyn oligomerization assays to show that Hsp70 directly blocks ASyn oligomerization, an early event in ASyn misfolding. Using truncations, mutations and inhibitors, we confirmed that Hsp70 interacts with ASyn via an as yet unidentified, non-canonical interaction site in the C-terminal domain. Finally, a biological role for a non-canonical interaction was observed in H4 neuroglioma cells. Together, these findings suggest that new chemical approaches will be required to target Hsp70-ASyn interaction in synucleinopathies. Such approaches are likely to be more specific than targeting Hsp70 canonical actions. Additionally, these results raise the question of whether other misfolded proteins might also engage via the same non-canonical mechanism.
]]></description>
<dc:creator>Tao, J.</dc:creator>
<dc:creator>Berthet, A.</dc:creator>
<dc:creator>Citron, R.</dc:creator>
<dc:creator>Stanley, R.</dc:creator>
<dc:creator>Gestwicki, J.</dc:creator>
<dc:creator>Agard, D. A.</dc:creator>
<dc:creator>McConlogue, L.</dc:creator>
<dc:date>2020-05-16</dc:date>
<dc:identifier>doi:10.1101/2020.05.15.098699</dc:identifier>
<dc:title><![CDATA[Hsp70 chaperone blocks α-synuclein oligomer formation via a novel engagement mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.20.105379v1?rss=1">
<title>
<![CDATA[
Massively multiplex single-molecule oligonucleosome footprinting 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.20.105379v1?rss=1</link>
<description><![CDATA[
Our understanding of the beads-on-a-string arrangement of nucleosomes has been built largely on high-resolution sequence-agnostic imaging methods and sequence-resolved bulk biochemical techniques. To bridge the divide between these approaches, we present the single-molecule adenine methylated oligonucleosome sequencing assay (SAMOSA). SAMOSA is a high-throughput single-molecule sequencing method that combines adenine methyltransferase footprinting and single-molecule real-time DNA sequencing to natively and nondestructively measure nucleosome positions on individual chromatin fibres. SAMOSA data allows unbiased classification of single-molecular  states of nucleosome occupancy on individual chromatin fibres. We leverage this to estimate nucleosome regularity and spacing on single chromatin fibres genome-wide, at predicted transcription factor binding motifs, and across both active and silent human epigenomic domains. Our analyses suggest that chromatin is comprised of a diverse array of both regular and irregular single-molecular oligonucleosome patterns that differ subtly in their relative abundance across epigenomic domains. This irregularity is particularly striking in constitutive heterochromatin, which has typically been viewed as a conformationally static entity. Our proof-of-concept study provides a powerful new methodology for studying nucleosome organization at a previously intractable resolution, and offers up new avenues for modeling and visualizing higher-order chromatin structure.

1-sentence summaryHigh-throughput single-molecule real-time footprinting of chromatin arrays reveals heterogeneous patterns of oligonucleosome occupancy.
]]></description>
<dc:creator>Abdulhay, N. J.</dc:creator>
<dc:creator>McNally, C. P.</dc:creator>
<dc:creator>Hsieh, L. J.</dc:creator>
<dc:creator>Kasinathan, S.</dc:creator>
<dc:creator>Keith, A.</dc:creator>
<dc:creator>Estes, L. S.</dc:creator>
<dc:creator>Karimzadeh, M.</dc:creator>
<dc:creator>Underwood, J. G.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:date>2020-05-22</dc:date>
<dc:identifier>doi:10.1101/2020.05.20.105379</dc:identifier>
<dc:title><![CDATA[Massively multiplex single-molecule oligonucleosome footprinting]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.21.109470v1?rss=1">
<title>
<![CDATA[
Structure and reconstitution of a hydrolase complex that releases peptidoglycan from the membrane after polymerization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.21.109470v1?rss=1</link>
<description><![CDATA[
Bacteria are surrounded by a peptidoglycan cell wall that is essential for their survival1. During cell wall assembly, a lipid-linked disaccharide-peptide precursor called Lipid II is polymerized and crosslinked to produce mature peptidoglycan. As Lipid II is polymerized, nascent polymers remain membrane-anchored at one end and the other end becomes crosslinked to the matrix2-4. A longstanding question is how bacteria release newly synthesized peptidoglycan strands from the membrane to complete the synthesis of mature peptidoglycan. Here we show that a Staphylococcus aureus cell wall hydrolase and a membrane protein containing eight transmembrane helices form a complex that acts as a peptidoglycan release factor. The complex cleaves nascent peptidoglycan internally to produce free oligomers as well as lipid-linked oligomers that can undergo further elongation. The polytopic membrane protein, which is similar to a eukaryotic CAAX protease, controls the length of these products. A 2.6 [A] resolution structure of the complex shows that the membrane protein scaffolds the hydrolase to orient its active site for cleavage of the glycan strand. We propose that this complex serves to detach newly-synthesized peptidoglycan polymer from the cell membrane to complete integration into the cell wall matrix.
]]></description>
<dc:creator>Schaefer, K.</dc:creator>
<dc:creator>Owens, T. W.</dc:creator>
<dc:creator>Page, J. E.</dc:creator>
<dc:creator>Santiago, M.</dc:creator>
<dc:creator>Kahne, D.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:date>2020-05-22</dc:date>
<dc:identifier>doi:10.1101/2020.05.21.109470</dc:identifier>
<dc:title><![CDATA[Structure and reconstitution of a hydrolase complex that releases peptidoglycan from the membrane after polymerization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.22.110304v1?rss=1">
<title>
<![CDATA[
Discovery of multiple anti-CRISPRs uncovers anti-defense gene clustering in mobile genetic elements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.22.110304v1?rss=1</link>
<description><![CDATA[
Many prokaryotes employ CRISPR-Cas systems to combat invading mobile genetic elements (MGEs). In response, some MGEs have evolved Anti-CRISPR (Acr) proteins to bypass this immunity, yet the diversity, distribution and spectrum of activity of this immune evasion strategy remain largely unknown. Here, we uncover 11 new type I anti-CRISPR genes encoded on numerous chromosomal and extrachromosomal mobile genetic elements within Enterobacteriaceae and Pseudomonas. Candidate genes were identified adjacent to anti-CRISPR associated gene 5 (aca5) and assayed against a panel of six type I systems: I-F (Pseudomonas, Pectobacterium, and Serratia), I-E (Pseudomonas and Serratia), and I-C (Pseudomonas), revealing the type I-F and/or I-E acr genes and a new aca (aca9). We find that acr genes not only associate with other acr genes, but also with inhibitors of distinct bacterial defense systems. These genomic regions appear to be "anti-defense islands", reminiscent of the clustered arrangement of "defense islands" in prokaryotic genomes. Our findings expand on the diversity of CRISPR-Cas inhibitors and reveal the potential exploitation of acr loci neighborhoods for identifying new anti-defense systems.
]]></description>
<dc:creator>Pinilla-Redondo, R.</dc:creator>
<dc:creator>Shehreen, S.</dc:creator>
<dc:creator>Marino, N. D.</dc:creator>
<dc:creator>Fagerlund, R. D.</dc:creator>
<dc:creator>Brown, C. M.</dc:creator>
<dc:creator>Soerensen, S. J.</dc:creator>
<dc:creator>Fineran, P. C.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2020-05-22</dc:date>
<dc:identifier>doi:10.1101/2020.05.22.110304</dc:identifier>
<dc:title><![CDATA[Discovery of multiple anti-CRISPRs uncovers anti-defense gene clustering in mobile genetic elements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.25.115287v1?rss=1">
<title>
<![CDATA[
Single cell analysis of human site-specific melanocyte differentiation and the decoding of developmental programs in melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.25.115287v1?rss=1</link>
<description><![CDATA[
In humans, epidermal melanocytes are responsible for skin pigmentation, defense against ultraviolet radiation, and the deadliest common skin cancer, melanoma. While there is substantial overlap in melanocyte development pathways between different model organisms, species dependent differences are frequent and the conservation of these processes in human skin remains unresolved1-3. Thus, the biology of developing and adult human melanocytes remains largely uncharacterized. Here, we used a single-cell enrichment and RNA-sequencing pipeline to study human epidermal melanocytes derived directly from skin, capturing transcriptomes across different anatomic sites, developmental age, sexes, and multiple skin tones. Using donor-matched skin from distinct volar and non-volar anatomic locations, we uncovered subpopulations of melanocytes exhibiting site-specific enrichment that occurs during gestation and persists through adulthood. In addition, we identified human melanocyte differentiation transcriptional programs that are distinct from gene signatures generated from model systems. Finally, we use these programs to define patterns of dedifferentiation that are predictive of melanoma prognosis. Overall, the characterization of human melanocytes fresh from skin revealed new subpopulations, human-specific transcriptional programs, and valuable insights into melanoma dedifferentiation.
]]></description>
<dc:creator>Belote, R. L.</dc:creator>
<dc:creator>Le, D.</dc:creator>
<dc:creator>Maynard, A.</dc:creator>
<dc:creator>Lang, U. E.</dc:creator>
<dc:creator>Sinclair, A.</dc:creator>
<dc:creator>Planells-Palop, V.</dc:creator>
<dc:creator>Baskin, L. S.</dc:creator>
<dc:creator>Tward, A. D.</dc:creator>
<dc:creator>Darmanis, S.</dc:creator>
<dc:creator>Judson-Torres, R. L.</dc:creator>
<dc:date>2020-05-28</dc:date>
<dc:identifier>doi:10.1101/2020.05.25.115287</dc:identifier>
<dc:title><![CDATA[Single cell analysis of human site-specific melanocyte differentiation and the decoding of developmental programs in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.26.117572v1?rss=1">
<title>
<![CDATA[
Cellular-scale silicon probes for high-density, precisely-localized neurophysiology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.26.117572v1?rss=1</link>
<description><![CDATA[
Neural implants with large numbers of electrodes have become an important tool for examining brain functions. However, these devices typically displace a large intracranial volume compared to the neurons they record. This large size limits the density of implants, provokes tissue reactions that degrade chronic performance, and impedes the ability to accurately visualize recording sites within intact circuits. Here we report next-generation silicon-based neural probes at cellular-scale (5x10{micro}m cross-section), with ultra-high-density packing (as little as 66{micro}m between shanks) and 64 or 256 closely-spaced recording sites per probe. We show that these probes can be inserted into superficial or deep brain structures and maintain high-quality spike recordings in freely behaving rats for many weeks. Finally, we demonstrate a slice-in-place approach for the precise registration of recording sites relative to nearby neurons and anatomical features, including striatal {micro}-opioid receptor patches. This scalable technology provides a valuable tool for examining information processing within neural circuits, and potentially for human brain-machine interfaces.
]]></description>
<dc:creator>Egert, D.</dc:creator>
<dc:creator>Pettibone, J. R.</dc:creator>
<dc:creator>Lemke, S.</dc:creator>
<dc:creator>Patel, P. R.</dc:creator>
<dc:creator>Caldwell, C. M.</dc:creator>
<dc:creator>Cai, D.</dc:creator>
<dc:creator>Ganguly, K.</dc:creator>
<dc:creator>Chestek, C. A.</dc:creator>
<dc:creator>Berke, J.</dc:creator>
<dc:date>2020-05-29</dc:date>
<dc:identifier>doi:10.1101/2020.05.26.117572</dc:identifier>
<dc:title><![CDATA[Cellular-scale silicon probes for high-density, precisely-localized neurophysiology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.29.120220v1?rss=1">
<title>
<![CDATA[
Complement factor C1q mediates chronic neuron loss and inflammation post-brain injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.29.120220v1?rss=1</link>
<description><![CDATA[
While traumatic brain injury (TBI) acutely disrupts the cortex, most TBI-related disabilities reflect secondary injuries that accrue over time. The thalamus is a likely site of secondary damage because of its reciprocal connections with the cortex. Using a mouse model of cortical injury that does not directly damage subcortical structures, we found a chronic increase in C1q expression specifically in the corticothalamic circuit. Increased C1q expression co-localized with neuron loss and chronic inflammation, and correlated with altered cortical rhythms. Blocking C1q counteracted most of these outcomes, suggesting that C1q is a disease modifier in TBI. Since the corticothalamic circuit is important for sensory processing, attention, cognition, and sleep, all of which can be impaired by TBI, this circuit could be a new target for treating TBI-related disabilities.
]]></description>
<dc:creator>Holden, S. S.</dc:creator>
<dc:creator>Aboubakr, O.</dc:creator>
<dc:creator>Higashikubo, B.</dc:creator>
<dc:creator>Cho, F. S.</dc:creator>
<dc:creator>Chang, A. H.</dc:creator>
<dc:creator>Morningstar, A.</dc:creator>
<dc:creator>Mathur, V.</dc:creator>
<dc:creator>Kuhn, L. J.</dc:creator>
<dc:creator>Suri, P.</dc:creator>
<dc:creator>Sankaranarayanan, S.</dc:creator>
<dc:creator>Andrews-Zwilling, Y.</dc:creator>
<dc:creator>Aronica, E.</dc:creator>
<dc:creator>Yednock, T.</dc:creator>
<dc:creator>Paz, J. T.</dc:creator>
<dc:date>2020-05-31</dc:date>
<dc:identifier>doi:10.1101/2020.05.29.120220</dc:identifier>
<dc:title><![CDATA[Complement factor C1q mediates chronic neuron loss and inflammation post-brain injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.31.126573v1?rss=1">
<title>
<![CDATA[
Scalp EEG interictal high frequency oscillations as an objective EEG biomarker of infantile spasms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.31.126573v1?rss=1</link>
<description><![CDATA[
ObjectiveTo investigate the diagnostic utility of high frequency oscillations (HFOs) via scalp electroencephalogram (EEG) in infantile spasms.

MethodsWe retrospectively analyzed interictal slow-wave sleep EEGs sampled at 2,000 Hz recorded from 30 consecutive patients who were suspected of having infantile spasms. We measured the rate of HFOs (80-500 Hz) and the strength of the cross-frequency coupling between HFOs and slow-wave activity (SWA) at 3-4 Hz and 0.5-1 Hz as quantified with modulation indices (MIs).

ResultsTwenty-three patients (77%) exhibited active spasms during the overnight EEG recording. Although the HFOs were detected in all children, increased HFO rate and MIs correlated with the presence of active spasms (p < 0.001 by HFO rate; p < 0.01 by MIs at 3-4 Hz; p = 0.02 by MIs at 0.5-1 Hz). The presence of active spasms was predicted by the logistic regression models incorporating HFO-related metrics (AUC: 0.80-0.98) better than that incorporating hypsarrhythmia (AUC: 0.61). The predictive performance of the best model remained favorable (87.5% accuracy) after a cross-validation procedure.

ConclusionsIncreased rate of HFOs and coupling between HFOs and SWA are associated with active epileptic spasms.

SignificanceScalp-recorded HFOs may serve as an objective EEG biomarker for active epileptic spasms.

HighlightsO_LIObjective analyses of scalp high frequency oscillations and its coupling with slow-wave activity in infantile spasms were feasible.
C_LIO_LIIncreased rate of high frequency oscillations and its coupling with slow-wave activity correlated with active epileptic spasms.
C_LIO_LIThe scalp high frequency oscillations were also detected in neurologically normal children (although at the low rate).
C_LI
]]></description>
<dc:creator>Nariai, H.</dc:creator>
<dc:creator>Hussain, S. A.</dc:creator>
<dc:creator>Bernardo, D.</dc:creator>
<dc:creator>Motoi, H.</dc:creator>
<dc:creator>Sonoda, M.</dc:creator>
<dc:creator>Kuroda, N.</dc:creator>
<dc:creator>Asano, E.</dc:creator>
<dc:creator>Nguyen, J.</dc:creator>
<dc:creator>Elashoff, D.</dc:creator>
<dc:creator>Sankar, R.</dc:creator>
<dc:creator>Bragin, A.</dc:creator>
<dc:creator>Staba, R. J.</dc:creator>
<dc:creator>Wu, J. Y.</dc:creator>
<dc:date>2020-06-01</dc:date>
<dc:identifier>doi:10.1101/2020.05.31.126573</dc:identifier>
<dc:title><![CDATA[Scalp EEG interictal high frequency oscillations as an objective EEG biomarker of infantile spasms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.03.130559v1?rss=1">
<title>
<![CDATA[
NMDA receptors control cortical axonal projections via EPHRIN-B/EPHB signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.03.130559v1?rss=1</link>
<description><![CDATA[
Callosal projections from primary somatosensory cortex (S1) are key for processing somatosensory inputs and integrating sensory-motor information. How the callosal innervation pattern in S1 is formed during early postnatal development is not clear. We found that the normal termination pattern of these callosal projections is disrupted in cortex specific NMDAR mutants. Rather than projecting selectively to the primary/secondary somatosensory cortex (S1/S2) border, axons were uniformly distributed throughout S1. In addition, the density of this projection increased over postnatal life until the mice died by P30. By combining genetic and antibody-mediated loss of function, we demonstrated that it is NR2B-containing NMDA receptors in target S1 that mediate this guidance phenotype, thus playing a central role in interhemispheric connectivity. Furthermore, we found that this function of NMDA receptors in callosal circuit formation is independent of ion channel function and instead depends on interactions with EPHRIN-B/EPHB. Thus, NMDAR cooperates with EPHRIN-B/EPHB in target S1 to control the formation callosal circuits by acting as an accessory axon guidance receptor modulating EPH-dependent repulsion.
]]></description>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Huynh, T.</dc:creator>
<dc:creator>Noguchi, H.</dc:creator>
<dc:creator>Bush, J.</dc:creator>
<dc:creator>Pleasure, S.</dc:creator>
<dc:date>2020-06-04</dc:date>
<dc:identifier>doi:10.1101/2020.06.03.130559</dc:identifier>
<dc:title><![CDATA[NMDA receptors control cortical axonal projections via EPHRIN-B/EPHB signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.03.132720v1?rss=1">
<title>
<![CDATA[
pHLARE: A Genetically Encoded Ratiometric Lysosome pH Biosensor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.03.132720v1?rss=1</link>
<description><![CDATA[
Many lysosome functions are determined by a lumenal pH of ~5.0, including the activity of resident acid-activated hydrolases. Lysosome pH (pHlys) is increased in neurodegenerative disorders and predicted to be decreased in cancers, making it a potential target for therapeutics to limit the progression of these diseases. Accurately measuring pHlys, however, is limited by currently used dyes that accumulate in multiple intracellular compartments and cannot be propagated in clonal cells for longitudinal studies or in vivo determinations. To resolve this limitation, we developed a genetically encoded ratiometric pHlys biosensor, pHLARE (pH Lysosomal Activity REporter), which localizes predominantly in lysosomes, has a dynamic range of pH 4.0 to 6.5, and can be stably expressed in cells. Using pHLARE we show decreased pHlys with inhibiting activity of the mammalian target of rapamycin complex 1 (mTORC1), in breast and pancreatic cancer cells compared with tissue-matched untransformed cells, and with the activated oncogene H-RasV12. pHLARE is a new tool to accurately measure pHlys, for improved understanding of lysosome dynamics that could be a promising therapeutic target.

Summary StatementMost lysosome functions require a low lumenal pH, which is dysregulated in many human diseases. We report a new genetically biosensor to accurately measure lysosome pH dynamics, which we use to show decreased lysosome pH in cancer cell lines.
]]></description>
<dc:creator>Webb, B. A.</dc:creator>
<dc:creator>Cook, J.</dc:creator>
<dc:creator>Wittmann, T.</dc:creator>
<dc:creator>Barber, D. L.</dc:creator>
<dc:date>2020-06-04</dc:date>
<dc:identifier>doi:10.1101/2020.06.03.132720</dc:identifier>
<dc:title><![CDATA[pHLARE: A Genetically Encoded Ratiometric Lysosome pH Biosensor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.04.134262v1?rss=1">
<title>
<![CDATA[
Human Induced Pluripotent Stem Cell Derived Sensory Neurons are Sensitive to the Neurotoxic Effects of Paclitaxel 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.04.134262v1?rss=1</link>
<description><![CDATA[
Chemotherapy-induced peripheral neuropathy (CIPN) is a dose-limiting adverse event associated with treatment with paclitaxel and other chemotherapeutic agents. The prevention and treatment of CIPN are limited by a lack of understanding of the molecular mechanisms underlying this toxicity. In the current study, a human induced pluripotent stem cell–derived sensory neuron (iPSC-SN) model was developed for the study of chemotherapy-induced neurotoxicity. The iPSC-SNs express proteins characteristic of nociceptor, mechanoreceptor and proprioceptor sensory neurons and show Ca2+ influx in response to capsaicin, α,β-meATP and glutamate. iPSC-SNs are relatively resistant to the cytotoxic effects of paclitaxel, with IC50 values of 38.1 μM (95% CI: 22.9 – 70.9 μM) for 48 hr exposure and 9.3 μM (95% CI: 5.7 – 16.5 μM) for 72 hr treatment. Paclitaxel causes dose- and time-dependent changes in neurite network complexity detected by βIII-tubulin staining and high content imaging. The IC50 for paclitaxel reduction of neurite area was 1.4 μM (95% CI: 0.3 - 16.9 μM) for 48 hr exposure and 0.6 μM (95% CI: 0.09 - 9.9 μM) for 72 hr exposure. Decreased mitochondrial membrane potential, slower movement of mitochondria down the neurites and changes in glutamate-induced neuronal excitability were also observed with paclitaxel exposure. The iPSC-SNs were also sensitive to docetaxel, vincristine and bortezomib. Collectively, these data support the use of iPSC-SNs for detailed mechanistic investigations of genes and pathways implicated in chemotherapy-induced neurotoxicity and the identification of novel therapeutic approaches for its prevention and treatment.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Xiong, C.</dc:creator>
<dc:creator>Chua, K. C.</dc:creator>
<dc:creator>Stage, T. B.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Altman-Merino, A.</dc:creator>
<dc:creator>Chan, D.</dc:creator>
<dc:creator>Saraf, K.</dc:creator>
<dc:creator>Ferracini, A. C.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:creator>Kroetz, D. L.</dc:creator>
<dc:date>2020-06-05</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.134262</dc:identifier>
<dc:title><![CDATA[Human Induced Pluripotent Stem Cell Derived Sensory Neurons are Sensitive to the Neurotoxic Effects of Paclitaxel]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.04.134692v1?rss=1">
<title>
<![CDATA[
Distinct genetic pathways define pre-leukemic and compensatory clonal hematopoiesis in Shwachman-Diamond syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.04.134692v1?rss=1</link>
<description><![CDATA[
Shwachman-Diamond syndrome (SDS) is an inherited bone marrow failure syndrome with predisposition to developing leukemia. We found that multiple independent somatic hematopoietic clones arise early in life, most commonly harboring heterozygous mutations in EIF6 or TP53. EIF6 mutations cause functional compensation for the germline deficiency by alleviating the SDS ribosome joining defect, improving translation, and reducing p53 activation. TP53 mutations decrease checkpoint activation without affecting ribosome assembly. We link development of leukemia with acquisition of biallelic TP53 alterations. Our results define distinct pathways of clonal selection driven by germline fitness constraint and provide a mechanistic framework for clinical surveillance.
]]></description>
<dc:creator>Kennedy, A. L.</dc:creator>
<dc:creator>Myers, K. C.</dc:creator>
<dc:creator>Bowman, J.</dc:creator>
<dc:creator>Gibson, C. J.</dc:creator>
<dc:creator>Camarda, N. D.</dc:creator>
<dc:creator>Furutani, E. M.</dc:creator>
<dc:creator>Muscato, G. M.</dc:creator>
<dc:creator>Klein, R. H.</dc:creator>
<dc:creator>Ballotti, K.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Harris, C. E.</dc:creator>
<dc:creator>Galvin, A.</dc:creator>
<dc:creator>Malsch, M.</dc:creator>
<dc:creator>Dale, D.</dc:creator>
<dc:creator>Gansner, J. M.</dc:creator>
<dc:creator>Nakano, T. A.</dc:creator>
<dc:creator>Bertuch, A.</dc:creator>
<dc:creator>Vlachos, A.</dc:creator>
<dc:creator>Lipton, J. M.</dc:creator>
<dc:creator>Castillo, P.</dc:creator>
<dc:creator>Connelly, J.</dc:creator>
<dc:creator>Churpek, J.</dc:creator>
<dc:creator>Edwards, J. R.</dc:creator>
<dc:creator>Hijiya, N.</dc:creator>
<dc:creator>Ho, R. H.</dc:creator>
<dc:creator>Hofmann, I.</dc:creator>
<dc:creator>Huang, J. N.</dc:creator>
<dc:creator>Keel, S.</dc:creator>
<dc:creator>Lamble, A.</dc:creator>
<dc:creator>Lau, B. W.</dc:creator>
<dc:creator>Norkin, M.</dc:creator>
<dc:creator>Stieglitz, E.</dc:creator>
<dc:creator>Stock, W.</dc:creator>
<dc:creator>Walkovich, K.</dc:creator>
<dc:creator>Boettcher, S.</dc:creator>
<dc:creator>Brendel, C.</dc:creator>
<dc:creator>Fleming, M. D.</dc:creator>
<dc:creator>Davies, S. M.</dc:creator>
<dc:creator>Weller, E. A.</dc:creator>
<dc:creator>Bahl, C.</dc:creator>
<dc:creator>Carter, S. L.</dc:creator>
<dc:creator>Shimamura, A.</dc:creator>
<dc:creator>Lindsley, R. C.</dc:creator>
<dc:date>2020-06-05</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.134692</dc:identifier>
<dc:title><![CDATA[Distinct genetic pathways define pre-leukemic and compensatory clonal hematopoiesis in Shwachman-Diamond syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.04.134965v1?rss=1">
<title>
<![CDATA[
Donor-derived mitochondrial DNA variant peptides elicit allo-specific immune response in transplant patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.04.134965v1?rss=1</link>
<description><![CDATA[
In stem cell transplant, mitochondrial DNA (mtDNA) nonsynonymous single nucleotide variants (SNVs) between donor and recipient (D-R) trigger alloimmune responses and transplant rejection. Whether similar alloimmune responses occurs in solid-organ transplantation remains unknown, particularly with the presence of human leukocyte antigen mismatches. This study characterized mtDNA SNVs between D-R of 163 human lung transplant pairs, and then, post-transplantation, assessed alloimmune responses against donor-derived mitochondrial peptides using ELISpot to measure interferon gamma (IFN{gamma}) release from recipients monocytes. We identified a median of 6 nonsynonymous mtDNA SNVs (Interquartile Range = 4 - 9) per D-R pair. SNVs were predominantly located at MT-CYB, MT-ATP6, and MT-ND3 genes. The number of SNVs was higher in D-R race non-concordant pairs than in race-concordant pairs. Donor-derived mitochondrial peptides triggered a 19.8-fold higher IFN{gamma} release compared to recipient-derived peptide. These findings were validated in heart transplantation and show that donor-derived mitochondrial peptides trigger allo-specific immune responses after transplantation.
]]></description>
<dc:creator>Marishta, A.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Jang, M. K.</dc:creator>
<dc:creator>Cuttin, K.</dc:creator>
<dc:creator>Jackson, A. M.</dc:creator>
<dc:creator>Luikart, H.</dc:creator>
<dc:creator>Deuse, T.</dc:creator>
<dc:creator>Khush, K. K.</dc:creator>
<dc:creator>Schrepfer, S.</dc:creator>
<dc:creator>Agbor-Enoh, S.</dc:creator>
<dc:creator>Valantine, H.</dc:creator>
<dc:date>2020-06-05</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.134965</dc:identifier>
<dc:title><![CDATA[Donor-derived mitochondrial DNA variant peptides elicit allo-specific immune response in transplant patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.08.139642v1?rss=1">
<title>
<![CDATA[
Satellite repeat transcripts modulate heterochromatin condensates and safeguard chromosome stability in mouse embryonic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.08.139642v1?rss=1</link>
<description><![CDATA[
Heterochromatin maintains genome integrity and function, and is organised into distinct nuclear domains. Some of these domains are proposed to form by phase separation through the accumulation of HP1. Mammalian heterochromatin contains noncoding major satellite repeats (MSR), which are highly transcribed in mouse embryonic stem cells (ESCs). Here, we report that MSR transcripts can drive the formation of HP1 droplets in vitro, and scaffold heterochromatin into dynamic condensates in ESCs, leading to the formation of large nuclear domains that are characteristic of pluripotent cells. Depleting MSR transcripts causes heterochromatin to transition into a more compact and static state. Unexpectedly, changing heterochromatins biophysical properties has severe consequences for ESCs, including chromosome instability and mitotic defects. These findings uncover an essential role for MSR transcripts in modulating the organisation and properties of heterochromatin to preserve genome stability. They also provide new insights into the processes that could regulate phase separation and the functional consequences of disrupting the properties of heterochromatin condensates.
]]></description>
<dc:creator>Novo, C. L.</dc:creator>
<dc:creator>Wong, E.</dc:creator>
<dc:creator>Hockings, C.</dc:creator>
<dc:creator>Poudel, C.</dc:creator>
<dc:creator>Sheekey, E.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Kaminski Schierle, G. S.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:date>2020-06-08</dc:date>
<dc:identifier>doi:10.1101/2020.06.08.139642</dc:identifier>
<dc:title><![CDATA[Satellite repeat transcripts modulate heterochromatin condensates and safeguard chromosome stability in mouse embryonic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.10.111203v1?rss=1">
<title>
<![CDATA[
Identification of compounds producing non-visual photosensation via TrpA1 in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.10.111203v1?rss=1</link>
<description><![CDATA[
ABSTRACTTRPA1 receptors sense chemical irritants, but they do not normally respond to light. Previous studies have identified compounds that confer photosensitivity onto vertebrate TRPA1. However, the pharmacology of TRPA1-mediated non-visual photosensation remains poorly understood. To identify novel compounds that affect this process, we screened a large chemical library for compounds that increased light-elicited motor activity in larval zebrafish. We found structurally diverse hit compounds that were photoreactive and produced specific behavioral phenotypes. A subset of these compounds required functional TRPA1 to produce behavioral phenotypes in vivo. These findings provide novel prototype compounds for controlling TRPA1 with light and improve our understanding of non-visual TRPA1-mediated photosensation.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Cheng, D.</dc:creator>
<dc:creator>McCarroll, M.</dc:creator>
<dc:creator>Taylor, J.</dc:creator>
<dc:creator>Wu, T. S.</dc:creator>
<dc:creator>KOKEL, D.</dc:creator>
<dc:date>2020-06-10</dc:date>
<dc:identifier>doi:10.1101/2020.06.10.111203</dc:identifier>
<dc:title><![CDATA[Identification of compounds producing non-visual photosensation via TrpA1 in zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.10.142893v1?rss=1">
<title>
<![CDATA[
Sentinel p16INK4a+ cells in the basement membrane form a reparative niche in the lung 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.10.142893v1?rss=1</link>
<description><![CDATA[
Senescent cells are recognized drivers of aging-related decline in organ function, but deciphering the biology of senescence in vivo has been hindered by the paucity of tools to track and isolate senescent cells in tissues1-4. Deleting senescent cells from transgenic murine models have demonstrated therapeutic benefits in numerous age-related diseases5-11, but the identity, behavior, and function of the senescent cells deleted in vivo remain elusive. We engineered an ultra-sensitive reporter of p16INK4a, a biomarker of senescence12, to isolate and track p16INK4a+ cells in vivo. Surprisingly, p16INK4a+ mesenchymal cells appear in the basement membrane adjacent to epithelial progenitors in the lung shortly after birth, and these cells demonstrate senescent characteristics in vivo and ex vivo. Transcriptomic analysis of p16INK4a+ mesenchymal cells from non-aged lungs demonstrates a transition to a secretory phenotype upon airway epithelial injury. Heterotypic 3D organoid assays show that injured p16INK4a+ mesenchymal cells enhance epithelial progenitor proliferation, and we identified EREG as a novel airway progenitor mitogen produced by the secretory p16INK4a+ mesenchymal cells. Mesenchymal-specific deletion of the p16INK4a gene abrogates features of senescence in vivo, but also attenuates normal epithelial repair. Thus, p16INK4a+ mesenchymal cells can act as sentinels for the airway epithelial stem cell niche, poised to transition to a senescence-associated secretory phenotype to support barrier repair. Our data identify possible cellular targets in vivo for a rapidly growing list of senolytic therapies, but also raises important questions about the hidden cost of targeting senescent cells present in normal organs.
]]></description>
<dc:creator>Reyes de Mochel, N.</dc:creator>
<dc:creator>Cheong, K. N.</dc:creator>
<dc:creator>Cassandras, M.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Krasilnikov, M.</dc:creator>
<dc:creator>Matatia, P.</dc:creator>
<dc:creator>Molofsky, A.</dc:creator>
<dc:creator>Campisi, J.</dc:creator>
<dc:creator>PENG, T.</dc:creator>
<dc:date>2020-06-10</dc:date>
<dc:identifier>doi:10.1101/2020.06.10.142893</dc:identifier>
<dc:title><![CDATA[Sentinel p16INK4a+ cells in the basement membrane form a reparative niche in the lung]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.15.151498v1?rss=1">
<title>
<![CDATA[
Mobile element warfare via CRISPR and anti-CRISPR in Pseudomonas aeruginosa 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.15.151498v1?rss=1</link>
<description><![CDATA[
Bacteria deploy multiple defense mechanisms to prevent the invasion of mobile genetic elements (MGEs). CRISPR-Cas systems use RNA-guided nucleases to target MGEs, which in turn produce anti-CRISPR (Acr) proteins that inactivate Cas protein effectors. The minimal component Type I-C CRISPR-Cas subtype is highly prevalent in bacteria, and yet a lack of a tractable in vivo model system has slowed its study, the identification of cognate Acr proteins, and thus our understanding of its true role in nature. Here, we describe MGE-MGE conflict between a mobile Pseudomonas aeruginosa Type I-C CRISPR-Cas system always encoded on pKLC102-like conjugative elements, which are large mobile islands, and seven new Type I-C anti-CRISPRs (AcrIF2*, AcrIC3-IC8) encoded by phages, other mobile islands, and transposons. The P. aeruginosa Type I-C system possesses a total of 300 non-redundant spacers (from 980 spacers total) across the 42 genomes analyzed, predominantly targeting P. aeruginosa phages. Of the seven new Type I-C anti-CRISPRs, all but one are highly acidic, and four have surprisingly broad inhibition activity, blocking multiple distantly related P. aeruginosa Type I CRISPR system subtypes (e.g. I-C and I-F, or I-C and I-E), including AcrIF2 (now, AcrIF2*), a previously described DNA mimic. Anti-type I-C activity of AcrIF2* was far more sensitive to mutagenesis of acidic residues in AcrIF2* than anti-type I-F activity, suggesting distinct binding mechanisms for this highly negatively charged protein. Five of the seven Acr proteins block DNA-binding, while the other two act downstream of DNA-binding, likely by preventing Cas3 recruitment or activity. For one such Cas3 inhibitor (AcrIC3), we identify a novel anti-CRISPR evasion strategy: a cas3-cas8 gene fusion, which also occurs in nature. Collectively, the Type I-C CRISPR spacer diversity and corresponding anti-CRISPR response, all occurring on Pseudomonas MGEs, demonstrates an active co-evolutionary battle between parasitic elements.
]]></description>
<dc:creator>Leon, L. M.</dc:creator>
<dc:creator>Park, A. E.</dc:creator>
<dc:creator>Borges, A. L.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2020-06-15</dc:date>
<dc:identifier>doi:10.1101/2020.06.15.151498</dc:identifier>
<dc:title><![CDATA[Mobile element warfare via CRISPR and anti-CRISPR in Pseudomonas aeruginosa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.17.156091v1?rss=1">
<title>
<![CDATA[
Pain and itch processing by subpopulations of molecularly diverse spinal and trigeminal projection neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.17.156091v1?rss=1</link>
<description><![CDATA[
A remarkable molecular and functional heterogeneity of the primary sensory neurons and dorsal horn interneurons transmits pain- and or itch-relevant information, but the molecular signature of the projection neurons that convey the messages to the brain is unclear. Here, using retro-TRAP (translating ribosome affinity purification) and RNA-seq we reveal extensive molecular diversity of spino- and trigeminoparabrachial projection neurons, which to date are almost exclusively defined by their expression of the neurokinin 1 receptor (NK1R). Among the many genes identified, we highlight distinct subsets of Cck+, Nptx2+, Nmb+, and Crh+ expressing projection neurons. By combining in situ hybridization of retrogradely labeled neurons with Fos-based assays we also demonstrate significant functional heterogeneity, including both convergence and segregation of pain- and itch-provoking inputs onto molecularly diverse subsets of NK1R- and non-NK1R-expressing projection neurons. The current study provides the first comprehensive investigation into the molecular profiles and functional properties of projection neuron subtypes.
]]></description>
<dc:creator>Wercberger, R.</dc:creator>
<dc:creator>Braz, J. M.</dc:creator>
<dc:creator>Weinrich, J. A.</dc:creator>
<dc:creator>Basbaum, A. I.</dc:creator>
<dc:date>2020-06-19</dc:date>
<dc:identifier>doi:10.1101/2020.06.17.156091</dc:identifier>
<dc:title><![CDATA[Pain and itch processing by subpopulations of molecularly diverse spinal and trigeminal projection neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.18.158469v1?rss=1">
<title>
<![CDATA[
Reward processing electrophysiology in schizophrenia: Effects of age and illness phase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.18.158469v1?rss=1</link>
<description><![CDATA[
BackgroundReward processing abnormalities may underlie characteristic pleasure and motivational impairments in schizophrenia. Some neural measures of reward processing show strong age-related modulation, highlighting the importance of considering age effects on reward sensitivity. We compared event-related potentials (ERPs) reflecting reward anticipation (stimulus-preceding negativity, SPN) and evaluation (reward positivity, RewP; late-positive potential, LPP) across individuals with schizophrenia (SZ) and healthy controls (HC), with an emphasis on examining effects of chronological age, brain age (i.e., predicted age based on neurobiological measures), and illness phase.

MethodsSubjects underwent EEG while completing a slot-machine task for which rewards were not dependent on performance accuracy, speed, or other preparatory demands. Slot-machine task EEG responses were compared between 54 SZ and 54 HC individuals, ages 19 to 65. Reward-related ERPs were analyzed with respect to chronological age, categorically-defined illness phase (early; ESZ versus chronic schizophrenia; CSZ), and were used to model brain age relative to chronological age.

ResultsIllness phase-focused analyses indicated there were no group differences in average SPN or RewP amplitudes. However, a group x reward outcome interaction revealed that ESZ differed from HC in later outcome processing, reflected by greater LPP responses following loss versus reward (a reversal of the HC pattern). While brain age estimates did not differ among groups, depressive symptoms in SZ were associated with older brain age estimates while controlling for negative symptoms.

ConclusionsESZ and CSZ did not differ from HC in reward anticipation or early outcome processing during a cognitively undemanding reward task, highlighting areas of preserved functioning. However, ESZ showed altered later reward outcome evaluation, pointing to selective reward deficits during the early illness phase of schizophrenia. Further, an association between ERP-derived brain age and depressive symptoms in SZ extends prior findings linking depression with reward-related ERP blunting. Taken together, both illness phase and age may impact reward processing in SZ, and brain aging may offer a promising, novel marker of reward dysfunction that warrants further study.
]]></description>
<dc:creator>Abram, S. V.</dc:creator>
<dc:creator>Roach, B.</dc:creator>
<dc:creator>Holroyd, C. B.</dc:creator>
<dc:creator>Paulus, M.</dc:creator>
<dc:creator>Ford, J.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>Fryer, S. L.</dc:creator>
<dc:date>2020-06-20</dc:date>
<dc:identifier>doi:10.1101/2020.06.18.158469</dc:identifier>
<dc:title><![CDATA[Reward processing electrophysiology in schizophrenia: Effects of age and illness phase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.19.160770v1?rss=1">
<title>
<![CDATA[
Human Prostate Cancer-Associated Macrophage Subtypes with Prognostic Potential Revealed by Single-cell Transcriptomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.19.160770v1?rss=1</link>
<description><![CDATA[
Macrophages in the tumor microenvironment are causally linked with prostate cancer development and progression, yet little is known about their composition in neoplastic human tissue. By performing single cell transcriptomic analysis of human prostate cancer resident macrophages, three distinct populations were identified in the diseased prostate. Unexpectedly, macrophages isolated from the tumor-adjacent site of the prostatectomy specimen were identical to those from the tumorous site. Markers associated with canonical M1 and M2 macrophage phenotypes were identifiable, however these were not the main factors defining unique subtypes. The genes selectively associated with each macrophage cluster were used to develop a gene signature which was highly associated with both recurrence-free and metastasis-free survival. These results highlight the relevance of tissue-specific macrophage subtypes in the tumour microenvironment for prostate cancer progression and demonstrates the utility of profiling single-cell transcriptomics in human tumor samples as a strategy to design gene classifiers for patient prognostication.
]]></description>
<dc:creator>Siefert, J. C.</dc:creator>
<dc:creator>Cioni, B.</dc:creator>
<dc:creator>Muraro, M. J.</dc:creator>
<dc:creator>Alshalalfa, M.</dc:creator>
<dc:creator>Vivie, J.</dc:creator>
<dc:creator>van der Poel, H.</dc:creator>
<dc:creator>Feng, F. Y.</dc:creator>
<dc:creator>Wessels, L.</dc:creator>
<dc:creator>Zwart, W.</dc:creator>
<dc:creator>Bergman, A. M.</dc:creator>
<dc:date>2020-06-20</dc:date>
<dc:identifier>doi:10.1101/2020.06.19.160770</dc:identifier>
<dc:title><![CDATA[Human Prostate Cancer-Associated Macrophage Subtypes with Prognostic Potential Revealed by Single-cell Transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.20.163097v1?rss=1">
<title>
<![CDATA[
Female reproductive tract has low concentration of SARS-CoV2 receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.20.163097v1?rss=1</link>
<description><![CDATA[
There has been significant concern regarding fertility and reproductive outcomes during the SARS-CoV2 pandemic. Recent data suggests a high concentration of SARS-Cov2 receptors, ACE2 or TMPRSS2, in nasal epithelium and cornea, which explains person-to-person transmission. We investigated the prevalence of SARS-CoV2 receptors among reproductive tissues by exploring the single-cell sequencing datasets from uterus, myometrium, ovary, fallopian tube, and breast epithelium. We did not detect significant expression of either ACE2 or TMPRSS2 in the normal human myometrium, uterus, ovaries, fallopian tube, or breast. Furthermore, none of the cell types in the female reproductive organs we investigated, showed the co-expression of ACE2 with proteases, TMPRSS2, Cathepsin B (CTSB), and Cathepsin L (CTSL) known to facilitate the entry of SARS2-CoV2 into the host cell. These results suggest that myometrium, uterus, ovaries, fallopian tube, and breast are unlikely to be susceptible to infection by SARS-CoV2. Our findings suggest that COVID-19 is unlikely to contribute to pregnancy-related adverse outcomes such as preterm birth, transmission of COVID-19 through breast milk, oogenesis and female fertility.
]]></description>
<dc:creator>Goad, J.</dc:creator>
<dc:creator>Rudolph, J.</dc:creator>
<dc:creator>Rajkovic, A.</dc:creator>
<dc:date>2020-06-22</dc:date>
<dc:identifier>doi:10.1101/2020.06.20.163097</dc:identifier>
<dc:title><![CDATA[Female reproductive tract has low concentration of SARS-CoV2 receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.24.169631v1?rss=1">
<title>
<![CDATA[
Increased baseline RASGRP1 signals Enhance Stem Cell Fitness during Native Hematopoiesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.24.169631v1?rss=1</link>
<description><![CDATA[
Oncogenic mutations in RAS genes, like KRASG12D or NRASG12D, trap Ras in the active state and cause myeloproliferative disorder and T cell leukemia (T-ALL) when induced in the bone marrow via Mx1CRE. The RAS exchange factor RASGRP1 is frequently overexpressed in T-ALL patients. In T-ALL cell lines overexpression of RASGRP1 increases flux through the RASGTP/RasGDP cycle. Here we expanded RASGRP1 expression surveys in pediatric T-ALL and generated a RoLoRiG mouse model crossed to Mx1CRE to determine the consequences of induced RASGRP1 overexpression in primary hematopoietic cells. RASGRP1-overexpressing, GFP-positive cells outcompeted wild type cells and dominated the peripheral blood compartment over time. RASGRP1 overexpression bestows gain-of-function colony formation properties to bone marrow progenitors in medium containing limited growth factors. RASGRP1 overexpression enhances baseline mTOR-S6 signaling in the bone marrow, but not in vitro cytokine-induced signals. In agreement with these mechanistic findings, hRASGRP1-ires-EGFP enhances fitness of stem- and progenitor- cells, but only in the context of native hematopoiesis. RASGRP1 overexpression is distinct from KRASG12D or NRASG12D, does not cause acute leukemia on its own, and leukemia virus insertion frequencies predict that RASGRP1 overexpression can effectively cooperate with lesions in many other genes to cause acute T cell leukemia.Competing Interest StatementConflict of interest disclosure:
Jeroen Roose is a co-founder and scientific advisor of Seal Biosciences, Inc. and on the scientific advisory committee for the Mark Foundation for Cancer Research. C.G.M. receives research funding from Loxo Oncology, Abbvie, and Pfizer, and speaking fees from Amgen.
View Full Text
]]></description>
<dc:creator>Karra, L.</dc:creator>
<dc:creator>Romero-Moya, D.</dc:creator>
<dc:creator>Ksionda, O.</dc:creator>
<dc:creator>Krush, M.</dc:creator>
<dc:creator>Gu, Z.</dc:creator>
<dc:creator>Mues, M.</dc:creator>
<dc:creator>Depeille, P.</dc:creator>
<dc:creator>Mullighan, C.</dc:creator>
<dc:creator>Roose, J.</dc:creator>
<dc:date>2020-06-25</dc:date>
<dc:identifier>doi:10.1101/2020.06.24.169631</dc:identifier>
<dc:title><![CDATA[Increased baseline RASGRP1 signals Enhance Stem Cell Fitness during Native Hematopoiesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.24.169920v1?rss=1">
<title>
<![CDATA[
Recording of DNA binding events during gut commensalism reveals the action of a repurposed Candida albicans regulatory network 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.24.169920v1?rss=1</link>
<description><![CDATA[
Candida albicans is a central fungal component of the human gut microbiota and an opportunistic pathogen. Two C. albicans transcription factors, Wor1 and Efg1, control its ability to colonize the mammalian gut. They are also master regulators of an epigenetic switch required for mating. Here, we show that six additional mating regulators influence gut commensalism. Using an adapted Calling Card-Seq protocol to record Candida transcription factor DNA binding events in the host, we validated these relationships during murine gut colonization. Finally, by comparing the in-host transcriptomes of regulatory mutants with enhanced vs. diminished commensal fitness, we identified a set of candidate commensalism effectors. These include Cht2, a GPI-linked chitinase whose gene is bound by Wor1, Czf1, and Efg1 in vivo and that we show to promote commensalism. We conclude that the network required for a C. albicans sexual switch is biochemically active in the host digestive tract, where it is repurposed to direct commensalism.
]]></description>
<dc:creator>Witchley, J. N.</dc:creator>
<dc:creator>Brimacombe, C. A.</dc:creator>
<dc:creator>Abon, N. A.</dc:creator>
<dc:creator>Noble, S.</dc:creator>
<dc:date>2020-06-24</dc:date>
<dc:identifier>doi:10.1101/2020.06.24.169920</dc:identifier>
<dc:title><![CDATA[Recording of DNA binding events during gut commensalism reveals the action of a repurposed Candida albicans regulatory network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.26.174185v1?rss=1">
<title>
<![CDATA[
Non-canonical activation of the ER stress sensor ATF6 by Legionella pneumophila effectors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.26.174185v1?rss=1</link>
<description><![CDATA[
The intracellular bacterial pathogen Legionella pneumophila (L.p.) secretes ~330 effector proteins into the host cell to sculpt an Endoplasmic Reticulum (ER)-derived replicative niche. We previously reported five L.p. effectors that inhibit IRE1, a key sensor of the homeostatic unfolded protein response (UPR) pathway. In this study, we discovered a subset of L.p. toxins that selectively activate the UPR sensor ATF6, resulting in its cleavage, nuclear translocation and target gene transcription without affecting other UPR sensors such as PERK. In a deviation from the conventional model, this L.p. dependent activation of ATF6 does not require its transport to the Golgi or its cleavage by the S1P/S2P proteases. We believe that our findings highlight the unique regulatory control that L.p. exerts upon the three UPR sensors and expand the repertoire of bacterial proteins that selectively perturb host homeostatic pathways.
]]></description>
<dc:creator>Ibe, N. U.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:date>2020-06-27</dc:date>
<dc:identifier>doi:10.1101/2020.06.26.174185</dc:identifier>
<dc:title><![CDATA[Non-canonical activation of the ER stress sensor ATF6 by Legionella pneumophila effectors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.28.176248v1?rss=1">
<title>
<![CDATA[
Phosphorylation modulates liquid-liquid phase separation of the SARS-CoV-2 N protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.28.176248v1?rss=1</link>
<description><![CDATA[
The nucleocapsid (N) protein of coronaviruses serves two major functions: compaction of the RNA genome in the virion and regulation of viral gene transcription in the infected cell1–3. The N protein contains two globular RNA-binding domains surrounded by regions of intrinsic disorder4. Phosphorylation of the central disordered region is required for normal viral genome transcription5,6, which occurs in a cytoplasmic structure called the replication transcription complex (RTC)7–11. It is not known how phosphorylation controls N protein function. Here we show that the N protein of SARS-CoV-2, together with viral RNA, forms biomolecular condensates12–15. Unmodified N protein forms partially ordered gel-like structures that depend on multivalent RNA-protein and protein-protein interactions. Phosphorylation reduces a subset of these interactions, generating a more liquid-like droplet. We speculate that distinct oligomeric states support the two functions of the N protein: unmodified protein forms a structured oligomer that is suited for nucleocapsid assembly, and phosphorylated protein forms a liquid-like compartment for viral genome processing. Inhibitors of N protein phosphorylation could therefore serve as antiviral therapy.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Carlson, C. R.</dc:creator>
<dc:creator>Asfaha, J. B.</dc:creator>
<dc:creator>Ghent, C. M.</dc:creator>
<dc:creator>Howard, C. J.</dc:creator>
<dc:creator>Hartooni, N.</dc:creator>
<dc:creator>Morgan, D. O.</dc:creator>
<dc:date>2020-06-29</dc:date>
<dc:identifier>doi:10.1101/2020.06.28.176248</dc:identifier>
<dc:title><![CDATA[Phosphorylation modulates liquid-liquid phase separation of the SARS-CoV-2 N protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.28.176412v1?rss=1">
<title>
<![CDATA[
The deubiquitinase Usp9x regulates PRC2-mediated chromatin reprogramming during mouse development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.28.176412v1?rss=1</link>
<description><![CDATA[
ABSTRACTPluripotent cells of the mammalian embryo undergo extensive chromatin rewiring to prepare for lineage commitment after implantation. Repressive H3K27me3, deposited by Polycomb Repressive Complex 2 (PRC2), is reallocated from large gene-distal blankets in pre-implantation embryos to mark promoters of developmental genes. The factors that mediate this global redistribution of H3K27me3 are unknown. Here we report a post-translational mechanism that destabilizes PRC2 to constrict H3K27me3 during lineage commitment. Using an auxin-inducible degron system, we show that the deubiquitinase Usp9x is required for mouse embryonic stem (ES) cell self-renewal. Usp9x-high ES cells have high PRC2 levels and bear a chromatin and transcriptional signature of the pre-implantation embryo, whereas Usp9x-low ES cells resemble the post-implantation, gastrulating epiblast. We show that Usp9x interacts with, deubiquitinates and stabilizes PRC2. Deletion of Usp9x in post-implantation embryos results in the derepression of genes that normally gain H3K27me3 after gastrulation, followed by the appearance of morphological abnormalities at E9.5, pointing to a recurrent link between Usp9x and PRC2 during development. Usp9x is a marker of “stemness” and is mutated in various neurological disorders and cancers. Our results unveil a Usp9x-PRC2 regulatory axis that is critical at peri-implantation and may be redeployed in other stem cell fate transitions and disease states.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Macrae, T. A.</dc:creator>
<dc:creator>Ramalho-Santos, M.</dc:creator>
<dc:date>2020-06-29</dc:date>
<dc:identifier>doi:10.1101/2020.06.28.176412</dc:identifier>
<dc:title><![CDATA[The deubiquitinase Usp9x regulates PRC2-mediated chromatin reprogramming during mouse development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.29.128439v1?rss=1">
<title>
<![CDATA[
Discovery of a Cellular Mechanism Regulating Transcriptional Noise 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.29.128439v1?rss=1</link>
<description><![CDATA[
Stochastic fluctuations in gene expression (‘noise’) are often considered detrimental but, in other fields, fluctuations are harnessed for benefit (e.g., ‘dither’ or amplification of thermal fluctuations to accelerate chemical reactions). Here, we find that DNA base-excision repair amplifies transcriptional noise, generating increased cellular plasticity and facilitating reprogramming. The DNA-repair protein Apex1 recognizes modified nucleoside substrates to amplify expression noise—while homeostatically maintaining mean levels of expression— for virtually all genes across the transcriptome. This noise amplification occurs for both naturally occurring base modifications and unnatural base analogs. Single-molecule imaging shows amplified noise originates from shorter, but more intense, transcriptional bursts that occur via increased DNA supercoiling which first impedes and then accelerates transcription, thereby maintaining mean levels. Strikingly, homeostatic noise amplification potentiates fate-conversion signals during cellular reprogramming. These data suggest a functional role for the observed occurrence of modified bases within DNA in embryonic development and disease.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Desai, R.</dc:creator>
<dc:creator>Hansen, M. M. K.</dc:creator>
<dc:creator>Martin, B.</dc:creator>
<dc:creator>Yu, C.</dc:creator>
<dc:creator>Ding, S.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:creator>Weinberger, L. S.</dc:creator>
<dc:date>2020-06-30</dc:date>
<dc:identifier>doi:10.1101/2020.06.29.128439</dc:identifier>
<dc:title><![CDATA[Discovery of a Cellular Mechanism Regulating Transcriptional Noise]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.30.179648v1?rss=1">
<title>
<![CDATA[
High-content Imaging-based Pooled CRISPR Screens in Mammalian Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.30.179648v1?rss=1</link>
<description><![CDATA[
CRISPR (clustered regularly interspaced short palindromic repeats) -based gene inactivation provides a powerful means of linking genes to particular cellular phenotypes. CRISPR-based screening has mostly relied upon using large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited to phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed a microscopy-based approach, which we name optical enrichment, to computationally select cells displaying a particular CRISPR-induced phenotype, mark them by photo-conversion of an expressed photo-activatable fluorescent protein, and then isolate the fluorescent cells using fluorescence-activated cell sorting (FACS). A plugin was developed for the open source software Manager to automate the phenotypic identification and photo-conversion of cells, allowing ~1.5 million individual cells to be screened in 8 hr. We used this approach to screen 6092 sgRNAs targeting 544 genes for their effects on nuclear size regulation and identified 14 bona fide hits. These results present a highly scalable approach to facilitate imaging-based pooled CRISPR screens.
]]></description>
<dc:creator>Yan, X.</dc:creator>
<dc:creator>Stuurman, N.</dc:creator>
<dc:creator>Ribeiro, S. A.</dc:creator>
<dc:creator>Tanenbaum, M. E.</dc:creator>
<dc:creator>Horlbeck, M. A.</dc:creator>
<dc:creator>Liem, C. R.</dc:creator>
<dc:creator>Jost, M.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:creator>Vale, R.</dc:creator>
<dc:date>2020-07-01</dc:date>
<dc:identifier>doi:10.1101/2020.06.30.179648</dc:identifier>
<dc:title><![CDATA[High-content Imaging-based Pooled CRISPR Screens in Mammalian Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.09.195057v1?rss=1">
<title>
<![CDATA[
Biomolecular condensates amplify mRNA decapping by coupling protein interactions with conformational changes in Dcp1/Dcp2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.09.195057v1?rss=1</link>
<description><![CDATA[
Cells organize biochemical processes into biological condensates. P-bodies are cytoplasmic condensates enriched in factors important for mRNA degradation. P-bodies have been identified as sites of both mRNA storage and decay, but how these opposing outcomes may be achieved in condensates is unresolved. A critical step in mRNA degradation is removal of the 5-7-methylguanosine cap by Dcp1/Dcp2, which is highly enriched in P-bodies. Dcp1/Dcp2 activity is repressed in condensates in vitro and requires the activator Edc3. Activation of decapping is amplified in condensates relative to the surrounding solution due to stabilization of an autoinhibited state in Dcp1/Dcp2. Edc3 couples a conformational change in the Dcp1/Dcp2 active site with alteration of the protein-protein interactions driving phase separation to activate decapping in condensates. The composition-dependent regulation of enzyme activity in condensates occurs over length scales ranging from microns to [A]ngstroms and may control the functional state of P-bodies and related phase-separated compartments.

HIGHLIGHTSO_LImRNA decapping in droplets is repressed
C_LIO_LICatalytically inert droplets are activated by a change in condensate composition
C_LIO_LIA switch in enzymatic activity requires a conformational change in condensates
C_LIO_LICondensates amplify enzyme activation compared to surrounding solution
C_LI
]]></description>
<dc:creator>Ryan W Tibble</dc:creator>
<dc:creator>Anaïs Depaix</dc:creator>
<dc:creator>Joanna Kowalska</dc:creator>
<dc:creator>Jacek Jemielity</dc:creator>
<dc:creator>John D Gross</dc:creator>
<dc:date>2020-07-09</dc:date>
<dc:identifier>doi:10.1101/2020.07.09.195057</dc:identifier>
<dc:title><![CDATA[Biomolecular condensates amplify mRNA decapping by coupling protein interactions with conformational changes in Dcp1/Dcp2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.21.214874v1?rss=1">
<title>
<![CDATA[
A Transcriptional Switch Governing Fibroblast Plasticity Underlies Reversibility of Chronic Heart Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.21.214874v1?rss=1</link>
<description><![CDATA[
In diseased organs, stress-activated signaling cascades alter chromatin, triggering broad shifts in transcription and cell state that exacerbate pathology. Fibroblast activation is a common stress response that worsens lung, liver, kidney and heart disease, yet its mechanistic basis remains poorly understood1,2. Pharmacologic inhibition of the BET family of transcriptional coactivators alleviates cardiac dysfunction and associated fibrosis, providing a tool to mechanistically interrogate maladaptive fibroblast states and modulate their plasticity as a potential therapeutic approach3-8. Here, we leverage dynamic single cell transcriptomic and epigenomic interrogation of heart tissue with and without BET inhibition to reveal a reversible transcriptional switch underlying stress-induced fibroblast activation. Transcriptomes of resident cardiac fibroblasts demonstrated robust and rapid toggling between the quiescent fibroblast and activated myofibroblast state in a manner that directly correlated with BET inhibitor exposure and cardiac function. Correlation of single cell chromatin accessibility with cardiac function revealed a novel set of reversibly accessible DNA elements that correlated with disease severity. Among the most dynamic elements was an enhancer regulating the transcription factor MEOX1, which was specifically expressed in activated myofibroblasts, occupied putative regulatory elements of a broad fibrotic gene program, and was required for TGF{beta}-induced myofibroblast activation. CRISPR interference of the most dynamic cis-element within the enhancer, marked by nascent transcription, prevented TGF{beta}-induced activation of Meox1. These findings identify MEOX1 as a central regulator of stress-induced myofibroblast activation associated with cardiac dysfunction. The plasticity and specificity of the BET-dependent regulation of MEOX1 in endogenous tissue fibroblasts provides new trans- and cis- targets for treating fibrotic disease.
]]></description>
<dc:creator>Alexanian, M.</dc:creator>
<dc:creator>Przytycki, P. F.</dc:creator>
<dc:creator>Micheletti, R.</dc:creator>
<dc:creator>Padmanabhan, A.</dc:creator>
<dc:creator>Ye, L.</dc:creator>
<dc:creator>Travers, J. G.</dc:creator>
<dc:creator>Gonzalez Teran, B.</dc:creator>
<dc:creator>Duan, Q.</dc:creator>
<dc:creator>Ranade, S. S.</dc:creator>
<dc:creator>Felix, F.</dc:creator>
<dc:creator>Linares-Saldana, R.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Andreoletti, G.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Ivey, K. N.</dc:creator>
<dc:creator>Jain, R.</dc:creator>
<dc:creator>McKinsey, T. A.</dc:creator>
<dc:creator>Rosenfeld, M. G.</dc:creator>
<dc:creator>Gifford, C.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Haldar, S. M.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:date>2020-07-22</dc:date>
<dc:identifier>doi:10.1101/2020.07.21.214874</dc:identifier>
<dc:title><![CDATA[A Transcriptional Switch Governing Fibroblast Plasticity Underlies Reversibility of Chronic Heart Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.22.216093v1?rss=1">
<title>
<![CDATA[
Ventral Tegmental Area GABA, glutamate, and glutamate-GABA neurons are heterogenous in their electrophysiological and pharmacological properties 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.22.216093v1?rss=1</link>
<description><![CDATA[
The ventral tegmental area (VTA) contains dopamine neurons intermixed with GABA-releasing (expressing vesicular GABA transporter, VGaT), glutamate-releasing (expressing vesicular glutamate transporter, VGluT2), and co-releasing (co-expressing VGaT and VGluT2) neurons. By delivering INTRSECT viral vectors into VTA of double vglut2-Cre/vgat-Flp transgenic mice, we targeted specific VTA cell populations for ex vivo recordings. We found that VGluT2+ VGaT- and VGluT2+ VGaT+ neurons on average had relatively hyperpolarized resting membrane voltage, greater rheobase, and lower spontaneous firing frequency compared to VGluT2- VGaT+ neurons, suggesting that VTA glutamate-releasing and glutamate-GABA co-releasing neurons require stronger excitatory drive to fire than GABA-releasing neurons. In addition, we detected expression of Oprm1mRNA (encoding  opioid receptors, MOR) in VGluT2+ VGaT- and VGluT2- VGaT+ neurons, and their hyperpolarization by the MOR agonist DAMGO. Collectively, we demonstrate the utility of the double transgenic mouse to access VTA glutamate, glutamate-GABA and GABA neurons, and show some electrophysiological heterogeneity among them.

Impact StatementSome physiological properties of VTA glutamate-releasing and glutamate-GABA co-releasing neurons are distinct from those of VTA GABA-releasing neurons. -opioid receptor activation hyperpolarizes some VTA glutamate-releasing and some GABA-releasing neurons.
]]></description>
<dc:creator>Miranda-Barrientos, J. A.</dc:creator>
<dc:creator>Chambers, I.</dc:creator>
<dc:creator>Mongia, S.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:creator>Wang, H.-L.</dc:creator>
<dc:creator>Mateo-Semidey, G. E.</dc:creator>
<dc:creator>Margolis, E. B.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Morales, M.</dc:creator>
<dc:date>2020-07-23</dc:date>
<dc:identifier>doi:10.1101/2020.07.22.216093</dc:identifier>
<dc:title><![CDATA[Ventral Tegmental Area GABA, glutamate, and glutamate-GABA neurons are heterogenous in their electrophysiological and pharmacological properties]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.22.216564v1?rss=1">
<title>
<![CDATA[
T cell subset-selective IL2RA enhancers shape autoimmune diabetes risk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.22.216564v1?rss=1</link>
<description><![CDATA[
The majority of genetic variants associated with complex human autoimmune diseases reside in enhancers1-3, non-coding regulatory elements that control gene expression. In contrast with variants that directly alter protein-coding sequences, enhancer variants are predicted to tune gene expression modestly and function in specific cellular contexts4, suggesting that small alterations in the functions of key immune cell populations are sufficient to shape disease risk. Here we tested this concept by experimentally perturbing distinct enhancers governing the high affinity IL-2 receptor alpha chain (IL2RA; also known as CD25). IL2RA is an immune regulator that promotes the pro- and anti-inflammatory functions of conventional T cells (Tconvs) and regulatory T cells (Tregs), respectively, and non-coding genetic variants in IL2RA have been linked to multiple autoimmune disorders4. We previously tiled across the IL2RA locus using CRISPR-activation and identified a stimulation-responsive element (CaRE4) with an enhancer that modestly affects the kinetics of IL2RA expression in Tconvs5. This enhancer is conserved across species and harbors a common human SNP associated with protection from Type 1 Diabetes (T1D)5,6. We now identified an additional conserved enhancer, termed CaRE3 enhancer, which modestly affected steady state IL2RA expression in regulatory T cells (Tregs). Despite their seemingly subtle impact on gene expression, the CaRE3 and CaRE4 enhancers had pronounced yet divergent effects on the incidence of diabetes in autoimmune prone animals. Deletion of the conserved CaRE4 enhancer completely protected against autoimmune diabetes even in animals treated with an immunostimulating anti-PD1 checkpoint inhibitor, whereas deletion of the CaRE3 enhancer accelerated spontaneous disease progression. Quantitative multiplexed imaging of the pancreatic lymph nodes (panLNs) revealed that each enhancer deletion preferentially affected the protein expression levels of IL2RA in activated Tconvs or Tregs, reciprocally tuning local competition for IL-2 input signals. In animals lacking the CaRE4 enhancer, skewed IL-2 signaling favored Tregs, increasing their local density around activated Tconvs to strongly suppress emergence of autoimmune effectors. By contrast, in animals lacking the CaRE3 enhancer, IL-2 signals were skewed towards activated Tconvs, promoting their escape from Treg control. Collectively, this work illustrates how subtle changes in gene regulation due to non-coding variation can significantly alter disease progression and how distinct enhancers controlling the same gene can have opposing effects on disease outcomes through cell type-selective activity.
]]></description>
<dc:creator>Simeonov, D. R.</dc:creator>
<dc:creator>Wong, H. S.</dc:creator>
<dc:creator>Cortez, J. T.</dc:creator>
<dc:creator>Young, A.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Nguyen, V.</dc:creator>
<dc:creator>Park, K.</dc:creator>
<dc:creator>Umhoefer, J.</dc:creator>
<dc:creator>Indart, A. C.</dc:creator>
<dc:creator>Woo, J. M.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>Germain, R. N.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2020-07-22</dc:date>
<dc:identifier>doi:10.1101/2020.07.22.216564</dc:identifier>
<dc:title><![CDATA[T cell subset-selective IL2RA enhancers shape autoimmune diabetes risk]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.22.216572v1?rss=1">
<title>
<![CDATA[
Machine intelligence identifies soluble TNFa as a therapeutic target for spinal cord injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.22.216572v1?rss=1</link>
<description><![CDATA[
Traumatic spinal cord injury (SCI) produces a complex syndrome that is expressed across multiple endpoints ranging from molecular and cellular changes to functional behavioral deficits. Effective therapeutic strategies for CNS injury are therefore likely to manifest multi-factorial effects across a broad range of biological and functional outcome measures. Thus, multivariate analytic approaches are needed to capture the linkage between biological and neurobehavioral outcomes. Injury-induced neuroinflammation (NI) presents a particularly challenging therapeutic target, since NI is involved in both degeneration and repair1,2. Here, we used big-data integration and large-scale analytics to examine a large dataset of preclinical efficacy tests combining 5 different blinded, fully counter-balanced treatment trials for different acute anti-inflammatory treatments for cervical spinal cord injury in rats. Multi-dimensional discovery, using topological data analysis3 (TDA) and principal components analysis (PCA) revealed that only one showed consistent multidimensional syndromic benefit: intrathecal application of recombinant soluble TNF receptor 1 (sTNFR1), which showed an inverse-U dose response efficacy. Using the optimal acute dose, we showed that clinically-relevant 90 min delayed treatment profoundly affected multiple biological indices of NI in the first 48 hrs after injury, including reduction in pro-inflammatory cytokines and gene expression of a coherent complex of acute inflammatory mediators and receptors. Further, a 90 min delayed bolus dose of sTNFR1 reduced the expression of NI markers in the chronic perilesional spinal cord, and consistently improved neurological function over 6 weeks post SCI. These results provide validation of a novel strategy for precision preclinical drug discovery that is likely to improve translation in the difficult landscape of CNS trauma, and confirm the importance of TNF signaling as a therapeutic target.
]]></description>
<dc:creator>Huie, J. R.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Kyritsis, N.</dc:creator>
<dc:creator>Pan, J.</dc:creator>
<dc:creator>Irvine, K.-A.</dc:creator>
<dc:creator>Nielson, J. L.</dc:creator>
<dc:creator>Schupp, P.</dc:creator>
<dc:creator>Oldham, M. C.</dc:creator>
<dc:creator>Gensel, J. C.</dc:creator>
<dc:creator>Lin, A.</dc:creator>
<dc:creator>Segal, M. R.</dc:creator>
<dc:creator>Ratan, R.</dc:creator>
<dc:creator>Bresnahan, J. C.</dc:creator>
<dc:creator>Beattie, M.</dc:creator>
<dc:date>2020-07-24</dc:date>
<dc:identifier>doi:10.1101/2020.07.22.216572</dc:identifier>
<dc:title><![CDATA[Machine intelligence identifies soluble TNFa as a therapeutic target for spinal cord injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.23.218768v1?rss=1">
<title>
<![CDATA[
An atlas of lamina-associated chromatin across thirteen human cell types reveals cell-type-specific and multiple subtypes of peripheral heterochromatin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.23.218768v1?rss=1</link>
<description><![CDATA[
Association of chromatin with lamin proteins at the nuclear periphery has emerged as a potential mechanism to coordinate cell type-specific gene expression and maintain cellular identity via gene silencing. Unlike many histone modifications and chromatin-associated proteins, lamin-associated domains (LADs) have yet to be mapped genome-wide in a diverse panel of human cell types, which has limited our understanding of the role peripheral chromatin plays in development and disease. To address this gap, we mapped LAMIN B1 (LB1) across twelve human cell types encompassing pluripotent stem cells, intermediate progenitors, and differentiated cells from all three germ layers. Integrative analyses of this atlas of peripheral chromatin with publicly available genomic data, as well as gene expression and repressive histone maps generated for this study, revealed that in all twelve cellular contexts lamin-associated chromatin is organized into at least two subtypes defined by differences in LB1 occupancy, gene expression, chromatin accessibility, transposable elements, replication timing, and radial positioning. Most genes gain or lose LB1 occupancy consistent with their cell type along developmental trajectories; however, we also identified examples where the enhancer, but not the gene body and promoter, change LAD state. Imaging of fluorescently labeled DNA in single cells validated these transitions and showed intermediate radial positioning of LADs that are gene dense, relatively accessible, and dynamic across development. This atlas represents the largest resource to date for peripheral chromatin organization studies.
]]></description>
<dc:creator>Keough, K. C.</dc:creator>
<dc:creator>Shah, P. P.</dc:creator>
<dc:creator>Wickramasinghe, N. M.</dc:creator>
<dc:creator>Dundes, C. E.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>Salomon, R. E. A.</dc:creator>
<dc:creator>Whalen, S.</dc:creator>
<dc:creator>Loh, K. M.</dc:creator>
<dc:creator>Dubois, N.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Jain, R.</dc:creator>
<dc:date>2020-07-24</dc:date>
<dc:identifier>doi:10.1101/2020.07.23.218768</dc:identifier>
<dc:title><![CDATA[An atlas of lamina-associated chromatin across thirteen human cell types reveals cell-type-specific and multiple subtypes of peripheral heterochromatin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.26.215020v1?rss=1">
<title>
<![CDATA[
Cell-free DNA as a biomarker for prostate cancer: elevated concentration and decreased fragment size 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.26.215020v1?rss=1</link>
<description><![CDATA[
PurposeProstate cancer is the most commonly diagnosed neoplasm in American men. Although existing biomarkers may detect localized prostate cancer, additional strategies are necessary for improving detection and identifying aggressive disease that may require further intervention. One promising, minimally invasive biomarker is cell-free DNA (cfDNA), which consist of short DNA fragments released into circulation by dying or lysed cells that may reflect underlying cancer. Here we investigated whether differences in cfDNA concentration and cfDNA fragment size could improve the sensitivity for detecting more advanced and aggressive prostate cancer.

Materials and MethodsThis study included 268 individuals: 34 healthy controls, 112 men with localized prostate cancer who underwent radical prostatectomy (RP), and 122 men with metastatic castration-resistant prostate cancer (mCRPC). Plasma cfDNA concentration and fragment size were quantified with the Qubit 3.0 and the 2100 Bioanalyzer. The potential relationship between cfDNA concentration or fragment size and localized or mCRPC prostate cancer was evaluated with descriptive statistics, logistic regression, and area under the curve analysis with cross-validation.

ResultsPlasma cfDNA concentrations were elevated in mCRPC patients in comparison to localized disease (OR5 ng/mL = 1.34, P = 0.027) or to being a control (OR5 ng/mL = 1.69, P = 0.034). Decreased average fragment size was associated with an increased risk of localized disease compared to controls (OR5bp = 0.77, P = 0.0008).

ConclusionThis study suggests that cfDNA concentration and average cfDNA fragment size may provide a quick, cost-effective approach to help determine which patients will benefit most from further screening and/or disease monitoring to help improve prostate cancer outcomes.
]]></description>
<dc:creator>Chen, E.</dc:creator>
<dc:creator>Cario, C. L.</dc:creator>
<dc:creator>Leong, L.</dc:creator>
<dc:creator>Lopez, K.</dc:creator>
<dc:creator>Marquez, C.</dc:creator>
<dc:creator>Chu, C.</dc:creator>
<dc:creator>Li, P. S.</dc:creator>
<dc:creator>Oropeza, E.</dc:creator>
<dc:creator>Tenggara, I.</dc:creator>
<dc:creator>Cowan, J.</dc:creator>
<dc:creator>Simko, J. P.</dc:creator>
<dc:creator>Chan, J. M.</dc:creator>
<dc:creator>Friedlander, T.</dc:creator>
<dc:creator>Wyatt, A. W.</dc:creator>
<dc:creator>Aggarwal, R.</dc:creator>
<dc:creator>Paris, P. L.</dc:creator>
<dc:creator>Carroll, P. R.</dc:creator>
<dc:creator>Feng, F.</dc:creator>
<dc:creator>Witte, J. S.</dc:creator>
<dc:date>2020-07-26</dc:date>
<dc:identifier>doi:10.1101/2020.07.26.215020</dc:identifier>
<dc:title><![CDATA[Cell-free DNA as a biomarker for prostate cancer: elevated concentration and decreased fragment size]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.29.222430v1?rss=1">
<title>
<![CDATA[
Automatic segmentation of dentate nuclei for microstructure assessment: example of application to temporal lobe epilepsy patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.29.222430v1?rss=1</link>
<description><![CDATA[
Dentate nuclei (DNs) segmentation is helpful for assessing their potential involvement in neurological diseases. Once DNs have been segmented, it becomes possible to investigate whether DNs they are microstructurally affected, through analysis of quantitative MRI parameters, such as the ones derived from diffusion weighted imaging (DWI). This study, therefore, aimed to develop a fully automated segmentation method using the non-DWI (b0) images from a DWI dataset to obtain DN masks inherently registered with parameter maps.

Three different automatic methods were applied to healthy subjects in order to segment the DNs: registration to SUIT (a spatially unbiased atlas template of the cerebellum and brainstem), OPAL (Optimized Patch Match for Label fusion) and CNN (Convolutional Neural Network). DNs manual segmentation was considered the gold standard. Results show that the segmentation obtained with SUIT has an average Dice Similarity Coefficient (DSC) of 0.4907{+/-}0.0793 between the automatic SUIT masks and the gold standard. A comparison with manual masks was also performed for OPAL (DSC = 0.7624 {+/-} 0.1786) and CNN (DSC = 0.8658 {+/-} 0.0255), showing a better performance when using CNN.

OPAL and CNN were optimised on heathy subjects data with high spatial resolution from the Human Connectome Project. The three methods were further used to segment the DNs of a subset of subjects affected by Temporal Lobe Epilepsy (TLE). This subset was derived from a 3T MRI research study which included DWI data acquired with a coarser resolution. In TLE dataset, SUIT performed similarly to using the HCP dataset, with a DSC = 0.4145 {+/-} 0.1023. Using TLE data, OPAL performed worse than using HCP data: after changing the probability threshold the DSC was 0.4522 {+/-} 0.1178.

CNN was able to extract the DNs using the TLE data without need for retraining and with a good DSC = 0.7368 {+/-} 0.0799. Statistical comparison of quantitative parameters derived from DWI analysis, as well as volumes of each DN, revealed altered and lateralised changes in TLE patients compared to healthy controls.

The proposed CNN is therefore a viable option for accurate extraction of DNs from b0 images of DWI data with different resolutions and acquired at different sites.
]]></description>
<dc:creator>Gaviraghi, M.</dc:creator>
<dc:creator>Giovanni Savini, G.</dc:creator>
<dc:creator>Castellazzi, G.</dc:creator>
<dc:creator>Palesi, F.</dc:creator>
<dc:creator>Rolandi, N.</dc:creator>
<dc:creator>Sacco, S.</dc:creator>
<dc:creator>Pichiecchio, A.</dc:creator>
<dc:creator>Mariani, V.</dc:creator>
<dc:creator>Tartara, E.</dc:creator>
<dc:creator>Tassi, L.</dc:creator>
<dc:creator>Vitali, P.</dc:creator>
<dc:creator>D'Angelo, E.</dc:creator>
<dc:creator>Gandini Wheeler-Kingshott, C. A. M.</dc:creator>
<dc:date>2020-07-30</dc:date>
<dc:identifier>doi:10.1101/2020.07.29.222430</dc:identifier>
<dc:title><![CDATA[Automatic segmentation of dentate nuclei for microstructure assessment: example of application to temporal lobe epilepsy patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.31.224600v1?rss=1">
<title>
<![CDATA[
α-Synuclein plasma membrane localization correlates with cellular phosphatidylinositol polyphosphate levels. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.31.224600v1?rss=1</link>
<description><![CDATA[
The Parkinsons disease protein -synuclein (Syn) promotes membrane fusion and fission by interacting with various negatively charged phospholipids. Despite postulated roles in endocytosis and exocytosis, plasma membrane (PM) interactions of Syn are poorly understood. Here, we show that phosphatidylinositol 4,5-bisphosphate (PIP2) and phosphatidylinositol 3,4,5-trisphosphate (PIP3), two highly acidic components of inner PM leaflets, mediate plasma membrane localization of endogenous pools of Syn in A2780, HeLa, SH-SY5Y and SK-MEL-2 cells. We demonstrate that Syn binds reconstituted PIP2-membranes in a helical conformation in vitro and that PIP2 synthesizing kinases and hydrolyzing phosphatases reversibly redistribute Syn in cells. We further delineate that Syn-PM targeting follows phosphoinositide-3 kinase (PI3K)-dependent changes of cellular PIP2 and PIP3 levels, which collectively suggests that phosphatidylinositol polyphosphates contribute to Syns cellular function(s) at the plasma membrane.
]]></description>
<dc:creator>Eichmann, C.</dc:creator>
<dc:creator>Jacob, R. S.</dc:creator>
<dc:creator>Dema, A.</dc:creator>
<dc:creator>Mercadante, D.</dc:creator>
<dc:creator>Selenko, P.</dc:creator>
<dc:date>2020-07-31</dc:date>
<dc:identifier>doi:10.1101/2020.07.31.224600</dc:identifier>
<dc:title><![CDATA[α-Synuclein plasma membrane localization correlates with cellular phosphatidylinositol polyphosphate levels.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.31.231746v1?rss=1">
<title>
<![CDATA[
Engineered ACE2 receptor traps potently neutralize SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.31.231746v1?rss=1</link>
<description><![CDATA[
An essential mechanism for SARS-CoV-1 and -2 infection begins with the viral spike protein binding to the human receptor protein angiotensin-converting enzyme II (ACE2). Here we describe a stepwise engineering approach to generate a set of affinity optimized, enzymatically inactivated ACE2 variants that potently block SARS-CoV-2 infection of cells. These optimized receptor traps tightly bind the receptor binding domain (RBD) of the viral spike protein and prevent entry into host cells. We first computationally designed the ACE2-RBD interface using a two-stage flexible protein backbone design process that improved affinity for the RBD by up to 12-fold. These designed receptor variants were affinity matured an additional 14-fold by random mutagenesis and selection using yeast surface display. The highest affinity variant contained seven amino acid changes and bound to the RBD 170-fold more tightly than wild-type ACE2. With the addition of the natural ACE2 collectrin domain and fusion to a human Fc domain for increased stabilization and avidity, the most optimal ACE2 receptor traps neutralized SARS-CoV-2 pseudotyped lentivirus and authentic SARS-CoV-2 virus with half-maximal inhibitory concentrations (IC50) in the 10-100 ng/ml range. Engineered ACE2 receptor traps offer a promising route to fighting infections by SARS-CoV-2 and other ACE2-utilizing coronaviruses, with the key advantage that viral resistance would also likely impair viral entry. Moreover, such traps can be pre-designed for viruses with known entry receptors for faster therapeutic response without the need for neutralizing antibodies isolated or generated from convalescent patients.
]]></description>
<dc:creator>Glasgow, A.</dc:creator>
<dc:creator>Glasgow, J. E.</dc:creator>
<dc:creator>Limonta, D.</dc:creator>
<dc:creator>Solomon, P.</dc:creator>
<dc:creator>Lui, I.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Nix, M. A.</dc:creator>
<dc:creator>Rettko, N. J.</dc:creator>
<dc:creator>Lim, S. A.</dc:creator>
<dc:creator>Zha, S.</dc:creator>
<dc:creator>Yamin, R.</dc:creator>
<dc:creator>Kao, K.</dc:creator>
<dc:creator>Rosenberg, O. S.</dc:creator>
<dc:creator>Ravetch, J. V.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Zhou, X. X.</dc:creator>
<dc:creator>Hobman, T. C.</dc:creator>
<dc:creator>Kortemme, T. K.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2020-08-01</dc:date>
<dc:identifier>doi:10.1101/2020.07.31.231746</dc:identifier>
<dc:title><![CDATA[Engineered ACE2 receptor traps potently neutralize SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.03.235622v1?rss=1">
<title>
<![CDATA[
Developing Cyclic Peptomers as Broad-Spectrum Gram negative Bacterial Type III Secretion System Inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.03.235622v1?rss=1</link>
<description><![CDATA[
Antibiotic resistant bacteria are an emerging global health threat. New antimicrobials are urgently needed. The injectisome type III secretion system (T3SS), required by dozens of Gram-negative bacteria for virulence but largely absent from non-pathogenic bacteria, is an attractive antimicrobial target. We previously identified synthetic cyclic peptomers, inspired by the natural product phepropeptin D, that inhibit protein secretion through the Yersinia Ysc and Pseudomonas aeruginosa Psc T3SSs, but do not inhibit bacterial growth. Here we describe identification of an isomer, 4EpDN, that is two-fold more potent (IC50 4 M) than its parental compound. Furthermore, 4EpDN inhibited the Yersinia Ysa and the Salmonella SPI-1 T3SSs, suggesting that this cyclic peptomer has broad efficacy against evolutionarily distant injectisome T3SSs. Indeed, 4EpDN strongly inhibited intracellular growth of Chlamydia trachomatis in HeLa cells, which requires the T3SS. 4EpDN did not inhibit the unrelated Twin arginine translocation (Tat) system, nor did it impact T3SS gene transcription. Moreover, although the injectisome and flagellar T3SSs are evolutionarily and structurally related, the 4EpDN cyclic peptomer did not inhibit secretion of substrates through the Salmonella flagellar T3SS, indicating that cyclic peptomers broadly but specifically target the injestisome T3SS. 4EpDN reduced the number of T3SS basal bodies detected on the surface of Y. enterocolitica, as visualized using a fluorescent derivative of YscD, an inner membrane ring with low homology to flagellar protein FliG. Collectively, these data suggest that cyclic peptomers specifically inhibit the injectisome T3SS from a variety of Gram-negative bacteria, possibly by preventing complete T3SS assembly.

IMPORTANCETraditional antibiotics target both pathogenic and commensal bacteria, resulting in a disruption of the microbiota, which in turn is tied to a number of acute and chronic diseases. The bacterial type III secretion system (T3SS) is an appendage used by many bacterial pathogens to establish infection, but is largely absent from commensal members of the microbiota. In this study, we identify a new derivative of the cyclic peptomer class of T3SS inhibitors. These compounds inhibit the T3SS of the nosocomial ESKAPE pathogen Pseudomonas aeruginosa and enteropathogenic Yersinia and Salmonella. The impact of cyclic peptomers is specific to the T3SS, as other bacterial secretory systems are unaffected. Importantly, cyclic peptomers completely block replication of Chlamydia trachomatis, the causative agent of genital, eye, and lung infections, in human cells, a process that requires the T3SS. Therefore, cyclic peptomers represent promising virulence blockers that can specifically disarm a broad spectrum of Gram-negative pathogens.
]]></description>
<dc:creator>Lam, H.</dc:creator>
<dc:creator>Lau, T.</dc:creator>
<dc:creator>Lentz, A.</dc:creator>
<dc:creator>Sherry, J.</dc:creator>
<dc:creator>Cabrera-Cortez, A.</dc:creator>
<dc:creator>Hug, K.</dc:creator>
<dc:creator>Engel, J.</dc:creator>
<dc:creator>Lokey, S.</dc:creator>
<dc:creator>Auerbach Stone, V.</dc:creator>
<dc:date>2020-08-04</dc:date>
<dc:identifier>doi:10.1101/2020.08.03.235622</dc:identifier>
<dc:title><![CDATA[Developing Cyclic Peptomers as Broad-Spectrum Gram negative Bacterial Type III Secretion System Inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.06.238915v1?rss=1">
<title>
<![CDATA[
Peptide Antidotes to SARS-CoV-2 (COVID-19) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.06.238915v1?rss=1</link>
<description><![CDATA[
The design of an immunogenic scaffold that serves a role in treating a pathogen, and can be rapidly and predictively modeled, has remained an elusive feat. Here, we demonstrate that SARS-BLOCK synthetic peptide scaffolds act as antidotes to SARS-CoV-2 spike protein-mediated infection of human ACE2-expressing cells. Critically, SARS-BLOCK peptides are able to potently and competitively inhibit SARS-CoV-2 S1 spike protein receptor binding domain (RBD) binding to ACE2, the main cellular entry pathway for SARS-CoV-2, while also binding to neutralizing antibodies against SARS-CoV-2. In order to create this potential therapeutic antidote-vaccine, we designed, simulated, synthesized, modeled epitopes, predicted peptide folding, and characterized behavior of a novel set of synthetic peptides. The biomimetic technology is modeled off the receptor binding motif of the SARS-CoV-2 coronavirus, and modified to provide enhanced stability and folding versus the truncated wildtype sequence. These novel peptides attain single-micromolar binding affinities for ACE2 and a neutralizing antibody against the SARS-CoV-2 receptor binding domain (RBD), and demonstrate significant reduction of infection in nanomolar doses. We also demonstrate that soluble ACE2 abrogates binding of RBD to neutralizing antibodies, which we posit is an essential immune-evasive mechanism of the virus. SARS-BLOCK is designed to "uncloak" the viral ACE2 coating mechanism, while also binding to neutralizing antibodies with the intention of stimulating a specific neutralizing antibody response. Our peptide scaffolds demonstrate promise for future studies evaluating specificity and sensitivity of immune responses to our antidote-vaccine. In summary, SARS-BLOCK peptides are a promising COVID-19 antidote designed to combine the benefits of a therapeutic and vaccine, effectively creating a new generation of prophylactic and reactive antiviral therapeutics whereby immune responses can be enhanced rather than blunted.
]]></description>
<dc:creator>Watson, A.</dc:creator>
<dc:creator>Ferreira, L. M. R.</dc:creator>
<dc:creator>Hwang, P.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Stroud, R.</dc:creator>
<dc:date>2020-08-06</dc:date>
<dc:identifier>doi:10.1101/2020.08.06.238915</dc:identifier>
<dc:title><![CDATA[Peptide Antidotes to SARS-CoV-2 (COVID-19)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.07.241943v1?rss=1">
<title>
<![CDATA[
Robust Estimation of Noise for Electromagnetic Brain Imaging with the Champagne Algorithm 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.07.241943v1?rss=1</link>
<description><![CDATA[
Robust estimation of the number, location, and activity of multiple correlated brain sources has long been a challenging task in electromagnetic brain imaging from M/EEG data, one that is significantly impacted by interference from spontaneous brain activity, sensor noise, and other sources of artifacts. Recently, we introduced the Champagne algorithm, a novel Bayesian inference algorithm that has shown tremendous success in M/EEG source reconstruction. Inherent to Champagne and most other related Bayesian reconstruction algorithms is the assumption that the noise covariance in sensor data can be estimated from "baseline" or "control" measurements. However, in many scenarios, such baseline data is not available, or is unreliable, and it is unclear how best to estimate the noise covariance. In this technical note, we propose several robust methods to estimate the contributions to sensors from noise arising from outside the brain without the need for additional baseline measurements. The incorporation of these methods for noise covariance estimation improves the robust reconstruction of complex brain source activity under high levels of noise and interference, while maintaining the performance features of Champagne. Specifically, we show that the resulting algorithm, Champagne with noise learning, is quite robust to initialization and is computationally efficient. In simulations, performance of the proposed noise learning algorithm is consistently superior to Champagne without noise learning. We also demonstrate that, even without the use of any baseline data, Champagne with noise learning is able to reconstruct complex brain activity with just a few trials or even a single trial, demonstrating significant improvements in source reconstruction for electromagnetic brain imaging.
]]></description>
<dc:creator>Cai, C.</dc:creator>
<dc:creator>Diwakar, M.</dc:creator>
<dc:creator>Hashemi, A.</dc:creator>
<dc:creator>Haufe, S.</dc:creator>
<dc:creator>Sekihara, K.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2020-08-07</dc:date>
<dc:identifier>doi:10.1101/2020.08.07.241943</dc:identifier>
<dc:title><![CDATA[Robust Estimation of Noise for Electromagnetic Brain Imaging with the Champagne Algorithm]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.08.238469v1?rss=1">
<title>
<![CDATA[
An ultra-high affinity synthetic nanobody blocks SARS-CoV-2 infection by locking Spike into an inactive conformation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.08.238469v1?rss=1</link>
<description><![CDATA[
Without an effective prophylactic solution, infections from SARS-CoV-2 continue to rise worldwide with devastating health and economic costs. SARS-CoV-2 gains entry into host cells via an interaction between its Spike protein and the host cell receptor angiotensin converting enzyme 2 (ACE2). Disruption of this interaction confers potent neutralization of viral entry, providing an avenue for vaccine design and for therapeutic antibodies. Here, we develop single-domain antibodies (nanobodies) that potently disrupt the interaction between the SARS-CoV-2 Spike and ACE2. By screening a yeast surface-displayed library of synthetic nanobody sequences, we identified a panel of nanobodies that bind to multiple epitopes on Spike and block ACE2 interaction via two distinct mechanisms. Cryogenic electron microscopy (cryo-EM) revealed that one exceptionally stable nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains (RBDs) locked into their inaccessible down-state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for SARS-CoV-2 Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains stability and function after aerosolization, lyophilization, and heat treatment. These properties may enable aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia, promising to yield a widely deployable, patient-friendly prophylactic and/or early infection therapeutic agent to stem the worst pandemic in a century.
]]></description>
<dc:creator>Schoof, M.</dc:creator>
<dc:creator>Faust, B.</dc:creator>
<dc:creator>Saunders, R. A.</dc:creator>
<dc:creator>Sangwan, S.</dc:creator>
<dc:creator>Rezelj, V. V.</dc:creator>
<dc:creator>Hoppe, N.</dc:creator>
<dc:creator>Boone, M.</dc:creator>
<dc:creator>Billesboelle, C.</dc:creator>
<dc:creator>Zimanyi, M.</dc:creator>
<dc:creator>Deshpande, I.</dc:creator>
<dc:creator>Liang, J.</dc:creator>
<dc:creator>Anand, A. A.</dc:creator>
<dc:creator>Dobzinski, N.</dc:creator>
<dc:creator>Zha, B. S.</dc:creator>
<dc:creator>Barsi-Rhyne, B.</dc:creator>
<dc:creator>Bleyy, V.</dc:creator>
<dc:creator>Barile-Hill, A. W.</dc:creator>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Simoneau, C. R.</dc:creator>
<dc:creator>Leon, K.</dc:creator>
<dc:creator>White, K. M.</dc:creator>
<dc:creator>Nock, S.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Ralston, C. Y.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Garcia-Sastre, A.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Vignuzzi, M.</dc:creator>
<dc:creator>Quantitative Biosciences Institute (QBI) Coronavirus Research Group Structural Biology Consortium,</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:date>2020-08-10</dc:date>
<dc:identifier>doi:10.1101/2020.08.08.238469</dc:identifier>
<dc:title><![CDATA[An ultra-high affinity synthetic nanobody blocks SARS-CoV-2 infection by locking Spike into an inactive conformation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.10.243329v1?rss=1">
<title>
<![CDATA[
Durable Control of HIV-1 Using a Staphylococcus aureus Cas9-Expressing Lentivirus Co-Targeting Viral Latency and Host Susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.10.243329v1?rss=1</link>
<description><![CDATA[
CRISPR/Cas9 gene editing has the potential to revolutionize the clinical management of HIV-1 infection, and may eliminate the need for antiretroviral therapy (ART). Current gene therapies attempt to either excise HIV-1 provirus or target HIV-1 entry receptors to prevent infection of new cells. Using a viral dynamic model, we determined that combining these two interventions, in the presence or absence of ART, significantly lowers the gene editing efficacy thresholds required to achieve an HIV-1 cure. To implement this dual-targeting approach, we engineered a single lentiviral vector that simultaneously targets multiple highly-conserved regions of the provirus and the host CXCR4 coreceptor, and developed a novel coculture system enabling real-time monitoring of latent infection, viral reactivation, and infection of new target cells. Simultaneous dual-targeting depleted HIV-1-infected cells with significantly greater potency than vectors targeting either virus or host independently, highlighting its potential as an HIV-1 cure strategy.
]]></description>
<dc:creator>Chavez, L. R.</dc:creator>
<dc:creator>Reddy, N. R.</dc:creator>
<dc:creator>Raymond, K. A.</dc:creator>
<dc:creator>Bouzidi, M. S.</dc:creator>
<dc:creator>Desai, S.</dc:creator>
<dc:creator>Dossani, Z. Y.</dc:creator>
<dc:creator>Sperber, H. S.</dc:creator>
<dc:creator>Theiler, J.</dc:creator>
<dc:creator>Korber, B.</dc:creator>
<dc:creator>Pillai, S. K.</dc:creator>
<dc:date>2020-08-10</dc:date>
<dc:identifier>doi:10.1101/2020.08.10.243329</dc:identifier>
<dc:title><![CDATA[Durable Control of HIV-1 Using a Staphylococcus aureus Cas9-Expressing Lentivirus Co-Targeting Viral Latency and Host Susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.12.247759v1?rss=1">
<title>
<![CDATA[
Regulated delivery controls Drosophila Hedgehog, Wingless and Decapentaplegic signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.12.247759v1?rss=1</link>
<description><![CDATA[
Morphogen signaling proteins disperse across tissues to activate signal transduction in target cells. We investigated dispersion of Hedgehog (Hh), Wingless (Wg), and Bone morphogenic protein homolog Decapentaplegic (Dpp) in the Drosophila wing imaginal disc, and found that delivery to targets is regulated. Cells take up <5% Hh produced, and neither amounts taken up nor extent of signaling changes under conditions of Hh production from 50-200% normal amounts. Similarly, cells take up <25% Wg produced, and variation in Wg production from 50-700% normal has no effect on amounts taken up or signaling. Similar properties were observed for Dpp. Wing disc-produced Hh signals to disc-associated tracheal and myoblast as well as an approximately equal number of disc cells, but the extent of signaling in the disc is unaffected by the presence or absence of the tracheal cells and myoblasts. These findings show that target cells do not take up signaling proteins from a common pool and that both the amount and destination of delivered morphogens are regulated..

SummaryThe extent of Hh, Wg, and Dpp signaling is independent of the amount of signal produced or the number of recipient cells.
]]></description>
<dc:creator>Kornberg, T. B.</dc:creator>
<dc:creator>Hatori, R.</dc:creator>
<dc:date>2020-08-13</dc:date>
<dc:identifier>doi:10.1101/2020.08.12.247759</dc:identifier>
<dc:title><![CDATA[Regulated delivery controls Drosophila Hedgehog, Wingless and Decapentaplegic signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.12.248526v1?rss=1">
<title>
<![CDATA[
Systematic evaluation of genome sequencing as a first-tier diagnostic test for prenatal and pediatric disorders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.12.248526v1?rss=1</link>
<description><![CDATA[
Current clinical guidelines recommend three genetic tests for the assessment of fetal structural anomalies: karyotype to detect microscopically-visible balanced and unbalanced chromosomal rearrangements, chromosomal microarray (CMA) to detect sub-microscopic copy number variants (CNVs), and exome sequencing (ES) to identify individual nucleotide changes in coding sequence. Advances in genome sequencing (GS) analysis suggest that it is poised to displace the sequential application of all three conventional tests to become a single diagnostic approach for the assessment of fetal structural anomalies. However, systematic benchmarking is required to assure that GS can capture the full mutational spectrum associated with fetal structural anomalies and to accurately quantify the added diagnostic yield of GS. We applied a novel GS analytic framework that included the discovery, filtration, and interpretation of nine classes of genomic variation to 7,195 individuals. We assessed the sensitivity of GS to detect diagnostic variants (pathogenic or likely pathogenic) from three standard-of-care tests using 1,612 autism spectrum disorder quartet families (ASD; n=6,448) with matched GS, ES, and CMA data, and validated these findings in 46 fetuses with a clinically reportable variant originally identified by karyotype, CMA, or ES. We then assessed the added diagnostic yield of GS in 249 trios (n=747) comprising a fetus with a structural anomaly detected by ultrasound and two unaffected parents that were pre-screened with a combination of all three standard-of-care tests. Across both cohorts, our GS analytic framework identified 98.2% of all diagnostic variants detected by standard-of-care tests, including 100% of those originally detected by CMA (n=88) and ES (n=61), as well as 78.6% (n=11/14) of the chromosomal rearrangements identified by karyotype. The diagnostic yield from GS was 7.8% across all 1,612 ASD probands, almost two-fold more than CMA (4.4%) and three-fold more than ES (3.0%). We also demonstrated that the yield of ES can approach that of GS when CNVs are captured with high sensitivity from exome data (7.4% vs. 7.8%, respectively). In 249 pre-screened fetuses with structural anomalies, GS provided an additional diagnostic yield of 0.4% beyond the combination of all three tests (karyotype, CMA, and ES). Applying our benchmarking results to existing data indicates that GS can achieve an overall diagnostic yield of 46.1% in unselected fetuses with fetal structural anomalies, providing an estimated 17.2% increase in diagnostic yield over karyotype, 14.1% over CMA, and 36.1% over ES when sequence variants are assessed, and 4.1% when CNVs are also identified from exome data. In this study we demonstrate that GS is sensitive to the detection of almost all pathogenic variation captured by karyotype, CMA, and ES, provides a superior diagnostic yield than any individual test by a wide margin, and contributes a modest increase in diagnostic yield beyond the combination of all three tests. We also outline several strategies to aid the interpretation of GS variants that are cryptic to conventional technologies, which we anticipate will be increasingly encountered as comprehensive variant identification from GS is performed. Taken together, these data suggest GS warrants consideration as a first-tier diagnostic approach for fetal structural anomalies.
]]></description>
<dc:creator>Lowther, C.</dc:creator>
<dc:creator>Valkanas, E.</dc:creator>
<dc:creator>Giordano, J. L.</dc:creator>
<dc:creator>Wang, H. Z.</dc:creator>
<dc:creator>Currall, B. B.</dc:creator>
<dc:creator>O'Keefe, K.</dc:creator>
<dc:creator>Collins, R. L.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Austin-Tse, C. A.</dc:creator>
<dc:creator>Evangelista, E.</dc:creator>
<dc:creator>Aggarwal, V.</dc:creator>
<dc:creator>Lucente, D.</dc:creator>
<dc:creator>Gauthier, L. D.</dc:creator>
<dc:creator>Tolonen, C.</dc:creator>
<dc:creator>Sahakian, N.</dc:creator>
<dc:creator>An, J.-Y.</dc:creator>
<dc:creator>Dong, S.</dc:creator>
<dc:creator>Norton, M. E.</dc:creator>
<dc:creator>MacKenzie, T.</dc:creator>
<dc:creator>Devlin, B.</dc:creator>
<dc:creator>Gilmore, K.</dc:creator>
<dc:creator>Powell, B.</dc:creator>
<dc:creator>Brandt, A.</dc:creator>
<dc:creator>Vetrini, F.</dc:creator>
<dc:creator>DiVito, M.</dc:creator>
<dc:creator>Goldstein, D. B.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>MacArthur, D. G.</dc:creator>
<dc:creator>Hodge, J. C.</dc:creator>
<dc:creator>O'Donnell-Luria, A.</dc:creator>
<dc:creator>Rehm, H.</dc:creator>
<dc:creator>Vora, N.</dc:creator>
<dc:creator>Levy, B.</dc:creator>
<dc:creator>Brand, H.</dc:creator>
<dc:creator>Wapner, R.</dc:creator>
<dc:creator>Talkowski, M. E.</dc:creator>
<dc:date>2020-08-13</dc:date>
<dc:identifier>doi:10.1101/2020.08.12.248526</dc:identifier>
<dc:title><![CDATA[Systematic evaluation of genome sequencing as a first-tier diagnostic test for prenatal and pediatric disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.17.253450v1?rss=1">
<title>
<![CDATA[
The conserved ER-transmembrane protein TMEM39 coordinates with vesicular COPII to promote collagen secretion and prevent ER stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.17.253450v1?rss=1</link>
<description><![CDATA[
Dysregulation of collagen production and secretion contributes to aging and tissue fibrosis of major organs. How premature collagen proteins in the endoplasmic reticulum (ER) route as specialized cargos for secretion remains to be fully elucidated. Here, we report that TMEM39, an ER-localized transmembrane protein, regulates production and secretory cargo trafficking of procollagen. We identify the C. elegans ortholog TMEM-39 from an unbiased RNAi screen and show that deficiency of tmem-39 leads to striking defects in cuticle collagen production and constitutively high ER stress response. RNAi knockdown of the tmem-39 ortholog in Drosophila causes similar defects in collagen secretion from fat body cells. The cytosolic domain of human TMEM39A binds to Sec23A, a vesicle coat protein that drives collagen secretion and vesicular trafficking. TMEM-39 regulation of collagen secretion is independent of ER stress response and autophagy. We propose that roles of TMEM-39 in collagen secretion and preventing ER stress are likely evolutionarily conserved.
]]></description>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Luo, S.</dc:creator>
<dc:creator>oliveirabarbosa, g.</dc:creator>
<dc:creator>Bai, M.</dc:creator>
<dc:creator>Kornberg, T.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2020-08-17</dc:date>
<dc:identifier>doi:10.1101/2020.08.17.253450</dc:identifier>
<dc:title><![CDATA[The conserved ER-transmembrane protein TMEM39 coordinates with vesicular COPII to promote collagen secretion and prevent ER stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.20.256487v1?rss=1">
<title>
<![CDATA[
4 β-hydroxycholesterol is a pro-lipogenic factor that promotes SREBP1c expression and activity through Liver X-receptor. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.20.256487v1?rss=1</link>
<description><![CDATA[
Oxysterols are oxidized derivatives of cholesterol that play signaling roles in lipid biosynthesis and homeostasis. Here we show that 4{beta}-hydroxycholesterol (4{beta}-HC), a liver and serum abundant oxysterol of poorly defined function, is a potent and selective inducer of the master lipogenic transcription factor, Sterol Regulatory Element Binding Protein 1c (SREBP1c), but not the related steroidogenic transcription factor SREBP2. Mechanistically, 4{beta}-HC acts as a putative agonist for Liver X receptor (LXR), a sterol sensor and transcriptional regulator previously linked to SREBP1c activation. Unique among the oxysterol agonists of LXR, 4{beta}-HC induced expression of the lipogenic program downstream of SREBP1c, and triggered de novo lipogenesis both in primary hepatocytes and in mouse liver. 4{beta}-HC-acted in parallel to insulin-PI3K-dependent signaling to stimulate triglyceride synthesis and lipid droplet accumulation. Thus, 4{beta}-HC is an endogenous regulator of de novo lipogenesis through the LXR-SREBP1c axis.
]]></description>
<dc:creator>Moldavski, O.</dc:creator>
<dc:creator>Zushin, P.-J. H.</dc:creator>
<dc:creator>Berdan, C. A.</dc:creator>
<dc:creator>van Eijkeren, R. J.</dc:creator>
<dc:creator>Jiang, X.</dc:creator>
<dc:creator>Qian, M.</dc:creator>
<dc:creator>Ory, D. S.</dc:creator>
<dc:creator>Covey, D. F.</dc:creator>
<dc:creator>Nomura, D.</dc:creator>
<dc:creator>Stahl, A.</dc:creator>
<dc:creator>Weiss, E.</dc:creator>
<dc:creator>Zoncu, R.</dc:creator>
<dc:date>2020-08-21</dc:date>
<dc:identifier>doi:10.1101/2020.08.20.256487</dc:identifier>
<dc:title><![CDATA[4 β-hydroxycholesterol is a pro-lipogenic factor that promotes SREBP1c expression and activity through Liver X-receptor.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.25.265561v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 infection of human iPSC-derived cardiac cells predicts novel cytopathic features in hearts of COVID-19 patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.25.265561v1?rss=1</link>
<description><![CDATA[
Although COVID-19 causes cardiac dysfunction in up to 25% of patients, its pathogenesis remains unclear. Exposure of human iPSC-derived heart cells to SARS-CoV-2 revealed productive infection and robust transcriptomic and morphological signatures of damage, particularly in cardiomyocytes. Transcriptomic disruption of structural proteins corroborated adverse morphologic features, which included a distinct pattern of myofibrillar fragmentation and numerous iPSC-cardiomyocytes lacking nuclear DNA. Human autopsy specimens from COVID-19 patients displayed similar sarcomeric disruption, as well as cardiomyocytes without DNA staining. These striking cytopathic features provide new insights into SARS-CoV-2 induced cardiac damage, offer a platform for discovery of potential therapeutics, and raise serious concerns about the long-term consequences of COVID-19.
]]></description>
<dc:creator>Perez-Bermejo, J. A.</dc:creator>
<dc:creator>Kang, S. S.</dc:creator>
<dc:creator>Rockwood, S. J.</dc:creator>
<dc:creator>Simoneau, C. R.</dc:creator>
<dc:creator>Joy, D. A.</dc:creator>
<dc:creator>Ramadoss, G. N.</dc:creator>
<dc:creator>Silva, A. C.</dc:creator>
<dc:creator>Flanigan, W. R.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Whitman, J. D.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:creator>McDevitt, T. C.</dc:creator>
<dc:date>2020-08-25</dc:date>
<dc:identifier>doi:10.1101/2020.08.25.265561</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 infection of human iPSC-derived cardiac cells predicts novel cytopathic features in hearts of COVID-19 patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.26.251611v1?rss=1">
<title>
<![CDATA[
Optimization and scaling of patient-derived brain organoids uncovers deep phenotypes of disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.26.251611v1?rss=1</link>
<description><![CDATA[
Cerebral organoids provide unparalleled access to human brain development in vitro. However, variability induced by current culture methodologies precludes using organoids as robust disease models. To address this, we developed an automated Organoid Culture and Assay (ORCA) system to support longitudinal unbiased phenotyping of organoids at scale across multiple patient lines. We then characterized organoid variability using novel machine learning methods and found that the contribution of donor, clone, and batch is significant and remarkably consistent over gene expression, morphology, and cell-type composition. Next, we performed multi-factorial protocol optimization, producing a directed forebrain protocol compatible with 96-well culture that exhibits low variability while preserving tissue complexity. Finally, we used ORCA to study tuberous sclerosis, a disease with known genetics but poorly representative animal models. For the first time, we report highly reproducible early morphological and molecular signatures of disease in heterozygous TSC+/- forebrain organoids, demonstrating the benefit of a scaled organoid system for phenotype discovery in human disease models.
]]></description>
<dc:creator>Shah, K.</dc:creator>
<dc:creator>Bedi, R.</dc:creator>
<dc:creator>Rogozhnikov, A.</dc:creator>
<dc:creator>Ramkumar, P.</dc:creator>
<dc:creator>Tong, Z.</dc:creator>
<dc:creator>Rash, B.</dc:creator>
<dc:creator>Stanton, M.</dc:creator>
<dc:creator>Sorokin, J.</dc:creator>
<dc:creator>Apaydin, C.</dc:creator>
<dc:creator>Batarse, A.</dc:creator>
<dc:creator>Bergamaschi, J.</dc:creator>
<dc:creator>Blattner, R.</dc:creator>
<dc:creator>Brown, S.</dc:creator>
<dc:creator>Bosshardt, A.</dc:creator>
<dc:creator>Castrillo, C.</dc:creator>
<dc:creator>Dang, B.</dc:creator>
<dc:creator>Drusinsky, S.</dc:creator>
<dc:creator>Enriquez, L.</dc:creator>
<dc:creator>Grayson, D.</dc:creator>
<dc:creator>Hilliard, J.</dc:creator>
<dc:creator>Hsu, P.-K.</dc:creator>
<dc:creator>Johnson, C.</dc:creator>
<dc:creator>Jones, R.</dc:creator>
<dc:creator>Lash, A.</dc:creator>
<dc:creator>Lee, E.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>McKay, A.</dc:creator>
<dc:creator>Mount, E.</dc:creator>
<dc:creator>Nicola, J.</dc:creator>
<dc:creator>Oumzil, I.</dc:creator>
<dc:creator>Paek, J.</dc:creator>
<dc:creator>Pascoe, D.</dc:creator>
<dc:creator>Piepho, A.</dc:creator>
<dc:creator>Poust, S.</dc:creator>
<dc:creator>Quang, D.</dc:creator>
<dc:creator>Schultz, M.</dc:creator>
<dc:creator>Sims, J.</dc:creator>
<dc:creator>Taylor, P.</dc:creator>
<dc:creator>Treiman, G.</dc:creator>
<dc:creator>Wueseke, O.</dc:creator>
<dc:creator>Young, N.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:creator>Flanzer, D.</dc:creator>
<dc:creator>Chao, D.</dc:creator>
<dc:creator>Skibinski, G.</dc:creator>
<dc:creator>Kato, S.</dc:creator>
<dc:creator>Escola, S.</dc:creator>
<dc:date>2020-08-27</dc:date>
<dc:identifier>doi:10.1101/2020.08.26.251611</dc:identifier>
<dc:title><![CDATA[Optimization and scaling of patient-derived brain organoids uncovers deep phenotypes of disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.31.275511v1?rss=1">
<title>
<![CDATA[
DYRK2 is a ciliary kinase involved 1 in vertebrate Hedgehog signal transduction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.31.275511v1?rss=1</link>
<description><![CDATA[
Primary cilia are organelles specialized for signaling. We previously defined the proteomes of sea urchin and sea anemone cilia to identify ciliary proteins that predate the origin of bilateria. This evolutionary perspective on cilia identified DYRK2, a kinase not been previously implicated in ciliary biology. We found that DYRK2 localizes to cilia and that loss of DYRK2 disrupts ciliary morphology. We also found that DYRK2 participates in ciliary Hh signal transduction, communicating between SMO and GLI transcription factors. Mutation of mouse Dyrk2 resulted in skeletal defects reminiscent of those caused by loss of Indian hedgehog (Ihh). Like Dyrk2 mutations, pharmacological inhibition of DYRK2 dysregulates ciliary length control and attenuates Hedgehog signaling. Thus, DYRK2 is required for ciliary morphology, for Hedgehog signaling in vitro, and for skeletal development. We propose that DYRK2 is part of the mechanism that transduces SMO to activate GLI transcription factors within cilia.
]]></description>
<dc:creator>Morante, N.</dc:creator>
<dc:creator>Sigg, M. A.</dc:creator>
<dc:creator>Strauskulage, L.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:date>2020-08-31</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.275511</dc:identifier>
<dc:title><![CDATA[DYRK2 is a ciliary kinase involved 1 in vertebrate Hedgehog signal transduction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.31.276295v1?rss=1">
<title>
<![CDATA[
Ciliary signaling-patterned smooth muscle drives tubular elongation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.31.276295v1?rss=1</link>
<description><![CDATA[
During development, many tubular organs undergo extensive longitudinal growth to reach their defined length, essential for their function, but how they lengthen is poorly understood. Here, we found that primary cilia are critical for the elongation of the small intestine and esophagus during murine embryonic development. More specifically, HH ligands produced by the epithelium signaled via cilia in the surrounding mesenchyme to pattern the smooth muscle. Like attenuated ciliary HH signaling, partial ablation of the smooth muscle reduced elongation, revealing an essential role for smooth muscle in longitudinal growth. Disruption of cilia, HH signaling or the smooth muscle reduced residual stress within the gut wall, indicating that smooth muscle contributes to the mechanical properties of the developing gut. Reducing residual stress decreased nuclear YAP, an effector of the mechanotransductive Hippo pathway. Removing YAP in the mesenchyme did not affect smooth muscle formation, but attenuated proliferation and elongation, demonstrating that YAP interprets smooth muscle-generated force to promote proliferation. Together, our results reveal that ciliary signaling directs the formation of the smooth muscle layer which, in turn, generates mechanical forces that activate YAP-mediated proliferation. As this interplay of biochemical and mechanical signals drives elongation of both the esophagus and small intestine, we propose that this mechanism may underlie tubular organ elongation generally.

HighlightsO_LIPrimary cilia are essential for the elongation of the small intestine and esophagus during embryonic development
C_LIO_LICiliary signaling patterns the smooth muscle in the developing intestine and esophagus
C_LIO_LIThe smooth muscle contributes to tissue mechanics
C_LIO_LISmooth muscle-generated strain activates YAP to drive longitudinal growth of the tubular organs
C_LI
]]></description>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Paivinen, P.</dc:creator>
<dc:creator>Xia, C.</dc:creator>
<dc:creator>Krup, A. L.</dc:creator>
<dc:creator>Makela, T. P.</dc:creator>
<dc:creator>Mostov, K. E.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:date>2020-09-01</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.276295</dc:identifier>
<dc:title><![CDATA[Ciliary signaling-patterned smooth muscle drives tubular elongation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.31.276428v1?rss=1">
<title>
<![CDATA[
Biological Noise in an Organelle Size Control System 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.31.276428v1?rss=1</link>
<description><![CDATA[
Analysis of fluctuation in organelle size provides a new way to probe the mechanisms of organelle size control systems. By analyzing cell-to-cell variation and within-cell fluctuations of flagellar length in Chlamydomonas, we show that the flagellar length control system exhibits both types of variation. Cell to cell variation is dominated by cell size, while within-cell variation results from dynamic fluctuations that are subject to a constraint, providing evidence for a homeostatic size control system. We analyzed a series of candidate genes affecting flagella and found that flagellar length variation is increased in mutations which increase the average flagellar length, an effect that we show is consistent with a theoretical model for flagellar length regulation based on length-dependent intraflagellar transport balanced by length-independent disassembly. Comparing the magnitude and time-scale of length fluctuations with simple models suggests that tubulin assembly is not directly coupled with IFT-mediated arrival and that observed fluctuations involve tubulin assembly and disassembly events involving large numbers of tubulin dimers. Cells with greater differences in their flagellar lengths show impaired swimming but improved gliding motility, raising the possibility that cells have evolved mechanisms to tune intrinsic noise in length. Taken together our results show that biological noise exists at the level of subcellular structures, with a corresponding effect on cell function, and can provide new insights into the mechanisms of organelle size control.
]]></description>
<dc:creator>Bauer, D.</dc:creator>
<dc:creator>Ishikawa, H.</dc:creator>
<dc:creator>Wemmer, K.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2020-09-01</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.276428</dc:identifier>
<dc:title><![CDATA[Biological Noise in an Organelle Size Control System]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.02.276550v1?rss=1">
<title>
<![CDATA[
miR-200 deficiency promotes lung cancer metastasis by activating cancer-associated fibroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.02.276550v1?rss=1</link>
<description><![CDATA[
Lung adenocarcinoma, the most prevalent lung cancer subtype, is characterized by its high propensity to metastasize. Despite the importance of metastasis in lung cancer mortality, its underlying cellular and molecular mechanisms remain largely elusive. Here, we identified miR-200 miRNAs as potent suppressors for lung adenocarcinoma metastasis. miR-200 expression is specifically repressed in mouse metastatic lung adenocarcinomas, and miR-200 decrease strongly correlates with poor patient survival. Consistently, deletion of mir-200c/141 in the KrasLSL-G12D/+; Trp53flox/flox lung adenocarcinoma mouse model significantly promoted metastasis, generating a desmoplastic tumor stroma highly reminiscent of metastatic human lung cancer. miR-200 deficiency in lung cancer cells promotes the proliferation and activation of adjacent cancer-associated fibroblasts (CAFs), which in turn elevates the metastatic potential of cancer cells. miR-200 regulates the functional interaction between cancer cells and CAFs, at least in part, by targeting Notch ligand Jagged1 and Jagged2 in cancer cells and inducing Notch activation in adjacent CAFs. Hence, the interaction between cancer cells and CAFs constitutes an essential mechanism to promote metastatic potential.
]]></description>
<dc:creator>Xue, B.</dc:creator>
<dc:creator>Chuang, C.-H.</dc:creator>
<dc:creator>Prosser, H. M.</dc:creator>
<dc:creator>Fuziwara, C. S.</dc:creator>
<dc:creator>Chan, C.</dc:creator>
<dc:creator>Sahasrabudhe, N.</dc:creator>
<dc:creator>Kühn, M.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Biton, A.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Wilkinson, J. E.</dc:creator>
<dc:creator>McManus, M. T.</dc:creator>
<dc:creator>Bradley, A.</dc:creator>
<dc:creator>Winslow, M. M.</dc:creator>
<dc:creator>Su, B.</dc:creator>
<dc:creator>He, L.</dc:creator>
<dc:date>2020-09-03</dc:date>
<dc:identifier>doi:10.1101/2020.09.02.276550</dc:identifier>
<dc:title><![CDATA[miR-200 deficiency promotes lung cancer metastasis by activating cancer-associated fibroblasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.05.284539v1?rss=1">
<title>
<![CDATA[
Characterizing tissue composition through combined analysis of single-cell morphologies and transcriptional states 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.05.284539v1?rss=1</link>
<description><![CDATA[
Advances in spatial transcriptomics technologies enable optical profiling of morphological and transcriptional modalities from the same cells within tissues. Here, we present multi-modal structured embedding (MUSE), an approach to deeply characterize tissue heterogeneity through analysis of combined image and transcriptional single-cell measurements. We demonstrate that MUSE can discover cellular subpopulations missed by either modality as well as compensate for modality-specific noise. MUSE identified biologically meaningful cellular subpopulations and stereotyped spatial patterning within heterogeneous mouse cortex brain tissues, profiled by seqFISH+ or STARmap technologies. MUSE provides a framework for combining multi-modal single-cell data to reveal deeper insights into the states, functions and organization of cells in complex biological tissues.
]]></description>
<dc:creator>Bao, F.</dc:creator>
<dc:creator>Deng, Y.</dc:creator>
<dc:creator>Wan, S.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Dai, Q.</dc:creator>
<dc:creator>Altschuler, S.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:date>2020-09-05</dc:date>
<dc:identifier>doi:10.1101/2020.09.05.284539</dc:identifier>
<dc:title><![CDATA[Characterizing tissue composition through combined analysis of single-cell morphologies and transcriptional states]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.06.283481v1?rss=1">
<title>
<![CDATA[
Multivariate pattern analysis of brain structure predicts functional outcome after auditory-based cognitive training interventions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.06.283481v1?rss=1</link>
<description><![CDATA[
BackgroundCognitive gains following cognitive training interventions (CT) are associated with improved functioning in people with schizophrenia (SCZ). However, considerable inter-individual variability is observed. Here, we evaluate the sensitivity of brain structural features to predict functional response to auditory-based cognitive training (ABCT) at a single subject level.

MethodsWe employed whole-brain multivariate pattern analysis (MVPA) with support vector machine (SVM) modeling to identify grey matter (GM) patterns that predicted  higher vs.  lower functioning after 40 hours of ABCT at the single subject level in SCZ patients. The generalization capacity of the SVM model was evaluated by applying the original model through an Out-Of-Sample Cross Validation analysis (OOCV) to unseen SCZ patients from an independent sample that underwent 50 hours of ABCT.

ResultsThe whole-brain GM volume-based pattern classification predicted  higher vs.  lower functioning at follow-up with a balanced accuracy (BAC) of 69.4% (sensitivity 72.2%, specificity 66.7%) as determined by nested cross-validation. The neuroanatomical model was generalizable to an independent cohort with a BAC of 62.1% (sensitivity 90.9%, specificity 33.3%).

ConclusionsIn particular, greater baseline GM volume in regions within superior temporal gyrus, thalamus, anterior cingulate and cerebellum -- predicted improved functioning at the single-subject level following ABCT in SCZ participants.
]]></description>
<dc:creator>Kambeitz-Ilankovic, L.</dc:creator>
<dc:creator>Vinogradov, S.</dc:creator>
<dc:creator>Wenzel, J.</dc:creator>
<dc:creator>Fisher, M.</dc:creator>
<dc:creator>Haas, S.</dc:creator>
<dc:creator>Betz, L.</dc:creator>
<dc:creator>Penzel, N.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Koutsouleris, N.</dc:creator>
<dc:creator>Subramaniam, K.</dc:creator>
<dc:date>2020-09-08</dc:date>
<dc:identifier>doi:10.1101/2020.09.06.283481</dc:identifier>
<dc:title><![CDATA[Multivariate pattern analysis of brain structure predicts functional outcome after auditory-based cognitive training interventions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.08.285346v1?rss=1">
<title>
<![CDATA[
Label-free imaging and classification of live P. falciparum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.08.285346v1?rss=1</link>
<description><![CDATA[
Manual microscopic inspection of fixed and stained blood smears has remained the gold standard for Plasmodium parasitemia analysis for over a century. Unfortunately, smear preparation consumes time and reagents, while manual microscopy is skill-dependent and labor-intensive. Here, we demonstrate that label-free microscopy combined with deep learning enables both life stage classification and accurate parasitemia quantification. Using a custom-built microscope, we find that deep-ultraviolet light enhances image contrast and resolution, achieving four-category classification of Plasmodium falciparum blood stages at an overall accuracy greater than 99%. To increase accessibility, we extended our method to a commercial brightfield microscope using near-ultraviolet and visible light. Both systems were tested extrinsically by parasitemia titration, revealing superior performance over manually-scored Giemsa-stained smears, and a limit of detection below 0.1%. Our results suggest that label-free microscopy combined with deep learning could eliminate the need for conventional blood smear analysis.
]]></description>
<dc:creator>Lebel, P. M.</dc:creator>
<dc:creator>Dial, R.</dc:creator>
<dc:creator>Vemuri, V. N. P.</dc:creator>
<dc:creator>Garcia, V. E.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:creator>Gomez-Sjoberg, R.</dc:creator>
<dc:date>2020-09-09</dc:date>
<dc:identifier>doi:10.1101/2020.09.08.285346</dc:identifier>
<dc:title><![CDATA[Label-free imaging and classification of live P. falciparum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.10.291872v1?rss=1">
<title>
<![CDATA[
Fine-tuning of Epithelial EGFR signals Supports Coordinated Mammary Gland Development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.10.291872v1?rss=1</link>
<description><![CDATA[
During puberty, robust morphogenesis occurs in the mammary gland; stem- and progenitor-cells develop into mature basal- and luminal-cells to form the ductal tree. The receptor signals that govern this process in mammary epithelial cells (MECs) are incompletely understood. The EGFR has been implicated and here we focused on EGFRs downstream pathway component Rasgrp1. We find that Rasgrp1 dampens EGF-triggered signals in MECs. Biochemically and in vitro, Rasgrp1 perturbation results in increased EGFR-Ras-PI3K-AKT and mTORC1-S6 kinase signals, increased EGF-induced proliferation, and aberrant branching-capacity in 3D cultures. However, in vivo, Rasgrp1 perturbation results in delayed ductal tree maturation with shortened branches and reduced cellularity. Rasgrp1-deficient MEC organoids revealed lower frequencies of basal cells, the compartment that incorporates stem cells. Molecularly, EGF effectively counteracts Wnt signal-driven stem cell gene signature in organoids. Collectively, these studies demonstrate the need for fine-tuning of EGFR signals to properly instruct mammary epithelium during puberty.
]]></description>
<dc:creator>Samocha, A.</dc:creator>
<dc:creator>Doh, H.</dc:creator>
<dc:creator>Sitarama, V.</dc:creator>
<dc:creator>Nguyen, Q.</dc:creator>
<dc:creator>Gbenedio, O.</dc:creator>
<dc:creator>Rudolf, J.</dc:creator>
<dc:creator>Eckalbar, W.</dc:creator>
<dc:creator>Barczak, A.</dc:creator>
<dc:creator>Miao, Y.</dc:creator>
<dc:creator>Garcia, C.</dc:creator>
<dc:creator>Lawson, D.</dc:creator>
<dc:creator>Werb, Z.</dc:creator>
<dc:creator>Kessenbrock, K.</dc:creator>
<dc:creator>Depeille, P.</dc:creator>
<dc:creator>Roose, J.</dc:creator>
<dc:date>2020-09-10</dc:date>
<dc:identifier>doi:10.1101/2020.09.10.291872</dc:identifier>
<dc:title><![CDATA[Fine-tuning of Epithelial EGFR signals Supports Coordinated Mammary Gland Development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.11.281782v1?rss=1">
<title>
<![CDATA[
Single-cell RNA sequencing of Tocilizumab-treated peripheral blood mononuclear cells as an in vitro model of inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.11.281782v1?rss=1</link>
<description><![CDATA[
COVID-19 has posed a significant threat to global health. Early data has revealed that IL-6, a key regulatory cytokine, plays an important role in the cytokine storm of COVID-19. Multiple trials are therefore looking at the effects of Tocilizumab, an IL-6 receptor antibody that inhibits IL-6 activity, on treatment of COVID-19, with promising findings. As part of a clinical trial looking at the effects of Tocilizumab treatment on kidney transplant recipients with subclinical rejection, we performed single-cell RNA sequencing of comparing stimulated PBMCs before and after Tocilizumab treatment. We leveraged this data to create an in vitro cytokine storm model, to better understand the effects of Tocilizumab in the presence of inflammation. Tocilizumab-treated cells had reduced expression of inflammatory-mediated genes and biologic pathways, particularly amongst monocytes. These results support the hypothesis that Tocilizumab may hinder the cytokine storm of COVID-19, through a demonstration of biologic impact at the single-cell level.
]]></description>
<dc:creator>Zarinsefat, A.</dc:creator>
<dc:creator>Hartoularos, G.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Sarwal, M. M.</dc:creator>
<dc:date>2020-09-11</dc:date>
<dc:identifier>doi:10.1101/2020.09.11.281782</dc:identifier>
<dc:title><![CDATA[Single-cell RNA sequencing of Tocilizumab-treated peripheral blood mononuclear cells as an in vitro model of inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.15.298083v1?rss=1">
<title>
<![CDATA[
Zika virus alters centrosome organization to suppress the innate immune response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.15.298083v1?rss=1</link>
<description><![CDATA[
Zika virus (ZIKV) is a flavivirus transmitted via mosquitoes and sex to cause congenital neurodevelopmental defects, including microcephaly. Inherited forms of microcephaly (MCPH) are associated with disrupted centrosome organization. Similarly, we found that ZIKV infection disrupted centrosome organization. ZIKV infection disrupted the organization of centrosomal proteins including CEP63, a MCPH-associated protein. The ZIKV nonstructural protein NS3 bound CEP63, and expression of NS3 was sufficient to alter centrosome architecture and CEP63 localization. Loss of CEP63 suppressed ZIKV-induced centrosome disorganization, indicating that ZIKV requires CEP63 to disrupt centrosome organization. ZIKV infection or loss of CEP63 decreased the centrosomal localization and stability of TANK-binding kinase 1 (TBK1), a regulator of the innate immune response. ZIKV infection or loss of CEP63 also increased the centrosomal accumulation of the CEP63 interactors, Mindbomb1 (MIB1) and DTX4, ubiquitin ligases that respectively activate and degrade TBK1. Therefore, we propose that ZIKV NS3 binds CEP63 to increase centrosomal DTX4 localization and destabilization of TBK1, thereby tempering the innate immune response. In addition to identifying a mechanism by which CEP63 controls the innate immune responses in ZIKV infection, we propose that the altered centrosomal organization caused by altered CEP63 function may contribute to ZIKV-associated microcephaly.
]]></description>
<dc:creator>Kodani, A.</dc:creator>
<dc:creator>Knopp, K.</dc:creator>
<dc:creator>Di Lullo, E.</dc:creator>
<dc:creator>Retallack, H.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:creator>Reiter, J.</dc:creator>
<dc:date>2020-09-15</dc:date>
<dc:identifier>doi:10.1101/2020.09.15.298083</dc:identifier>
<dc:title><![CDATA[Zika virus alters centrosome organization to suppress the innate immune response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.16.297945v1?rss=1">
<title>
<![CDATA[
Characterisation of protease activity during SARS-CoV-2 infection identifies novel viral cleavage sites and cellular targets for drug repurposing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.16.297945v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 is the causative agent behind the COVID-19 pandemic, and responsible for over 170 million infections, and over 3.7 million deaths worldwide. Efforts to test, treat and vaccinate against this pathogen all benefit from an improved understanding of the basic biology of SARS-CoV-2. Both viral and cellular proteases play a crucial role in SARS-CoV-2 replication, and inhibitors targeting proteases have already shown success at inhibiting SARS-CoV-2 in cell culture models. Here, we study proteolytic cleavage of viral and cellular proteins in two cell line models of SARS-CoV-2 replication using mass spectrometry to identify protein neo-N-termini generated through protease activity. We identify previously unknown cleavage sites in multiple viral proteins, including major antigenic proteins S and N, which are the main targets for vaccine and antibody testing efforts. We discovered significant increases in cellular cleavage events consistent with cleavage by SARS-CoV-2 main protease, and identify 14 potential high-confidence substrates of the main and papain-like proteases, validating a subset with in vitro assays. We showed that siRNA depletion of these cellular proteins inhibits SARS-CoV-2 replication, and that drugs targeting two of these proteins: the tyrosine kinase SRC and Ser/Thr kinase MYLK, showed a dose-dependent reduction in SARS-CoV-2 titres. Overall, our study provides a powerful resource to understand proteolysis in the context of viral infection, and to inform the development of targeted strategies to inhibit SARS-CoV-2 and treat COVID-19.
]]></description>
<dc:creator>Meyer, B.</dc:creator>
<dc:creator>Chiaravalli, J.</dc:creator>
<dc:creator>Brownridge, P.</dc:creator>
<dc:creator>Bryne, D. P.</dc:creator>
<dc:creator>Daly, L. A.</dc:creator>
<dc:creator>Agou, F.</dc:creator>
<dc:creator>Eyers, C. E.</dc:creator>
<dc:creator>Eyers, P. A.</dc:creator>
<dc:creator>Vignuzzi, M.</dc:creator>
<dc:creator>Emmott, E.</dc:creator>
<dc:date>2020-09-16</dc:date>
<dc:identifier>doi:10.1101/2020.09.16.297945</dc:identifier>
<dc:title><![CDATA[Characterisation of protease activity during SARS-CoV-2 infection identifies novel viral cleavage sites and cellular targets for drug repurposing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.18.296913v1?rss=1">
<title>
<![CDATA[
The CD28-transmembrane domain mediates chimeric antigen receptor heterodimerization with CD28 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.18.296913v1?rss=1</link>
<description><![CDATA[
Anti-CD19 chimeric antigen receptor (CD19-CAR)-engineered T cells are approved therapeutics for malignancies. The impact of the hinge (HD) and transmembrane (TMD) domains between the extracellular antigen-targeting and the intracellular signaling modalities of CARs has not been systemically studied. Here, a series of CD19-CARs differing only by their HD (CD8/CD28/IgG4) and TMD (CD8/CD28) was generated. CARs containing a CD28-TMD, but not a CD8-TMD, formed heterodimers with the endogenous CD28 in human T cells, as shown by co-immunoprecipitation and CAR-dependent proliferation to anti-CD28 stimulation. This dimerization depended on polar amino-acids in the CD28-TMD. CD28-CAR heterodimerization was more efficient in CARs containing a CD8-HD or CD28-HD as compared to an IgG4-HD. CD28-CAR heterodimers did not respond to CD80 and CD86 stimulation but led to a significant reduction of CD28 cell-surface expression. These data unveil a new property of the CD28-TMD and suggest that TMDs can modulate CAR T-cell activities by engaging endogenous partners.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=118 SRC="FIGDIR/small/296913v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Muller, Y.</dc:creator>
<dc:creator>Nguyen, D.</dc:creator>
<dc:creator>Ferreira, L. M. R.</dc:creator>
<dc:creator>Ho, P.</dc:creator>
<dc:creator>Raffin, C.</dc:creator>
<dc:creator>Beltran Valencia, R.</dc:creator>
<dc:creator>Congrave-Wilson, Z.</dc:creator>
<dc:creator>Roth, T.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Van Gool, F.</dc:creator>
<dc:creator>Marson, A. M. R.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:creator>Bluestone, J. A.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:date>2020-09-19</dc:date>
<dc:identifier>doi:10.1101/2020.09.18.296913</dc:identifier>
<dc:title><![CDATA[The CD28-transmembrane domain mediates chimeric antigen receptor heterodimerization with CD28]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.26.314997v1?rss=1">
<title>
<![CDATA[
Dscam1 promotes blood cell survival in Drosophila melanogaster through a dual role in blood cells and neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.26.314997v1?rss=1</link>
<description><![CDATA[
Down Syndrome Cell Adhesion Molecule 1 (Dscam1) is a receptor-like cell adhesion molecule that is conserved across the animal kingdom, but its roles in hematopoiesis remain unknown. Dscam1 related genes in vertebrates and invertebrates are key regulators of neuron morphogenesis and neuronal tiling. In Drosophila, Dscam1 in addition has roles in blood cells (hemocytes) in innate immunity and phagocytosis of pathogens. Given the anatomical and functional role of peripheral sensory neurons as microenvironments for resident hematopoietic sites in the Drosophila larva, we sought to investigate the role of Dscam1 in this context. Interestingly, we find that Dscam1 fills the role of a previously anticipated factor in neuron-hemocyte communication that supports trophic survival: tissue specific silencing of Dscam1 by in vivo RNAi in sensory neurons leads to neuron reduction, which in turn results in reduced hemocyte numbers due to apoptosis. Dscam1 silencing in hemocytes also results in a reduction of hemocytes and increased apoptosis. This cell-autonomous effect of Dscam1 silencing can be mimicked by RNAi silencing of dreadlocks (dock), suggesting that intracellular Dscam1 signaling relies on the adapter protein Dock in this system. Our findings reveal a dual role for Dscam1 in Drosophila hematopoiesis, by promoting survival of the sensory neuron microenvironments that in turn support hemocyte survival, and by promoting survival of hemocytes cell-autonomously. It will be interesting to explore possible functions of vertebrate Dscam1 related genes such as DSCAML1 in blood cells and their trophic survival.
]]></description>
<dc:creator>Ouyang, D.</dc:creator>
<dc:creator>Xiao, X.</dc:creator>
<dc:creator>Mase, A.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Corcoran, S.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Brueckner, K.</dc:creator>
<dc:date>2020-09-26</dc:date>
<dc:identifier>doi:10.1101/2020.09.26.314997</dc:identifier>
<dc:title><![CDATA[Dscam1 promotes blood cell survival in Drosophila melanogaster through a dual role in blood cells and neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.29.318501v1?rss=1">
<title>
<![CDATA[
Direct observation of genome surveillance by CRISPR-Cas in bacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.29.318501v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas systems are prokaryotic adaptive immune systems that have been well characterized biochemically, but in vivo spatiotemporal regulation and cell biology remains largely unaddressed. Here, we used fluorescent fusion proteins to study the localization of the Type I-F CRISPR-Cas system native to Pseudomonas aeruginosa. When targeted to an integrated prophage, the crRNA-guided (Csy) complex and a majority of Cas3 molecules in the cell are recruited to a single focus. When lacking a target in the cell, however, the Csy complex is broadly nucleoid bound, while Cas3 is diffuse in the cytoplasm. Nucleoid association for the Csy proteins is crRNA-dependent, and inhibited by expression of anti-CRISPR AcrIF2, which blocks PAM binding. The Cas9 nuclease is also nucleoid localized, only when gRNA-bound, which is abolished by PAM mimic, AcrIIA4. Our findings reveal PAM-dependent nucleoid surveillance and spatiotemporal regulation in Type I CRISPR-Cas that separates the nuclease-helicase Cas3 from the crRNA-guided surveillance complex.
]]></description>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Borges, A.</dc:creator>
<dc:creator>Govindarajan, S.</dc:creator>
<dc:date>2020-09-29</dc:date>
<dc:identifier>doi:10.1101/2020.09.29.318501</dc:identifier>
<dc:title><![CDATA[Direct observation of genome surveillance by CRISPR-Cas in bacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.30.321190v1?rss=1">
<title>
<![CDATA[
Early growth response 2 (EGR2) is a novel regulator of the senescence program 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.30.321190v1?rss=1</link>
<description><![CDATA[
Senescence, a state of stable growth arrest, plays an important role in ageing and age-related diseases in vivo. Although the INK4/ARF locus is known to be essential for senescence programs, the key regulators driving p16 and ARF transcription remain largely underexplored. Using siRNA screening for modulators of the p16/pRB and ARF/p53/p21 pathways in deeply senescent human mammary epithelial cells (DS HMECs) and fibroblasts (DS HMFs), we identified EGR2 as a novel regulator of senescence. EGR2 expression is up-regulated during senescence and its ablation by siRNA in DS HMECs and HMFs transiently reverses the senescent phenotype. We demonstrate that EGR2 activates the ARF and p16 promoters and directly binds to the ARF promoter. Loss of EGR2 downregulates p16 levels and increases the pool of p16- p21-  reversed cells in the population. Moreover, EGR2 overexpression is sufficient to induce senescence. Our data suggest that EGR2 is a regulator of the p16/pRB and direct transcriptional activator of the ARF/p53/p21 pathways in senescence and a novel marker of senescence.
]]></description>
<dc:creator>Tyler, E. J.</dc:creator>
<dc:creator>Gutierrez Del Arroyo, A.</dc:creator>
<dc:creator>Wallis, R.</dc:creator>
<dc:creator>Hughes, B. K.</dc:creator>
<dc:creator>Garbe, J. C.</dc:creator>
<dc:creator>Stampfer, M. R.</dc:creator>
<dc:creator>Koh, J.</dc:creator>
<dc:creator>Lowe, R.</dc:creator>
<dc:creator>Philpott, M. P.</dc:creator>
<dc:creator>Bishop, C. L.</dc:creator>
<dc:date>2020-10-01</dc:date>
<dc:identifier>doi:10.1101/2020.09.30.321190</dc:identifier>
<dc:title><![CDATA[Early growth response 2 (EGR2) is a novel regulator of the senescence program]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.02.323238v1?rss=1">
<title>
<![CDATA[
South Asian patient population genetics reveal strong founder effects and high rates of homozygosity - new resources for precision medicine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.02.323238v1?rss=1</link>
<description><![CDATA[
Population-scale genetic studies can identify drug targets and allow disease risk to be predicted with resulting benefit for management of individual health risks and system-wide allocation of health care delivery resources. Although population-scale projects are underway in many parts of the world, genetic variation between population groups means that additional projects are warranted. South Asia has a population whose genetics is the least characterized of any of the worlds major populations. Here we describe GenomeAsia studies that characterize population structure in South Asia and that create tools for economical and accurate genotyping at population-scale. Prior work on population structure characterized isolated population groups, the relevance of which to large-scale studies of disease genetics is unclear. For our studies we used whole genome sequence information from 4,807 individuals recruited in the health care delivery systems of Pakistan, India and Bangladesh to ensure relevance to population-scale studies of disease genetics. We combined this with WGS data from 927 individuals from isolated South Asian population groups, and developed a custom SNP array (called SARGAM) that is optimized for future human genetic studies in South Asia. We find evidence for high rates of reproductive isolation, endogamy and consanguinity that vary across the subcontinent and that lead to levels of homozygosity that approach 100 times that seen in outbred populations. We describe founder effects that increase the power to associate functional variants with disease processes and that make South Asia a uniquely powerful place for population-scale genetic studies.
]]></description>
<dc:creator>Wall, J.</dc:creator>
<dc:creator>Sathirapongsasuti, J. F.</dc:creator>
<dc:creator>Gupta, R.</dc:creator>
<dc:creator>Barik, A.</dc:creator>
<dc:creator>Rai, R. K.</dc:creator>
<dc:creator>Rasheed, A.</dc:creator>
<dc:creator>Radha, V.</dc:creator>
<dc:creator>Belsare, S.</dc:creator>
<dc:creator>Menon, R.</dc:creator>
<dc:creator>Phalke, S.</dc:creator>
<dc:creator>Mittal, A.</dc:creator>
<dc:creator>Fang, J.</dc:creator>
<dc:creator>Tanneeru, D.</dc:creator>
<dc:creator>Robinson, J.</dc:creator>
<dc:creator>Chaudhary, R.</dc:creator>
<dc:creator>Fuchsberger, C.</dc:creator>
<dc:creator>Forer, L.</dc:creator>
<dc:creator>Schoenherr, S.</dc:creator>
<dc:creator>Bei, Q.</dc:creator>
<dc:creator>Bhangale, T.</dc:creator>
<dc:creator>Tom, J.</dc:creator>
<dc:creator>Gadde, S. G. K.</dc:creator>
<dc:creator>V, P. B.</dc:creator>
<dc:creator>Naik, N. K.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Kwok, P.-Y.</dc:creator>
<dc:creator>Khera, A. V.</dc:creator>
<dc:creator>Lakshmi, B. R.</dc:creator>
<dc:creator>Butterworth, A.</dc:creator>
<dc:creator>Danesh, J.</dc:creator>
<dc:creator>Seshagiri, S.</dc:creator>
<dc:creator>Kathiresan, S.</dc:creator>
<dc:creator>Ghosh, A.</dc:creator>
<dc:creator>Mohan, V.</dc:creator>
<dc:creator>Chowdhury, A.</dc:creator>
<dc:creator>Saleheen, D.</dc:creator>
<dc:creator>Stawiski, E.</dc:creator>
<dc:creator>Peterson, A. S.</dc:creator>
<dc:date>2020-10-02</dc:date>
<dc:identifier>doi:10.1101/2020.10.02.323238</dc:identifier>
<dc:title><![CDATA[South Asian patient population genetics reveal strong founder effects and high rates of homozygosity - new resources for precision medicine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.03.319137v1?rss=1">
<title>
<![CDATA[
Role for the Na+/K+ ATPase α 3 subunit in folding and lamination of the human neocortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.03.319137v1?rss=1</link>
<description><![CDATA[
Osmotic equilibrium and membrane potential in animal cells depend on concentration gradients of sodium (Na+) and potassium (K+) ions across the plasma membrane, a function that is catalyzed by the Na,K-ATPase alpha subunit. In vertebrates, four paralogous genes, ATP1A1-4, encode distinct alpha subunit isoforms (a1-a4), three of which (a1, a2, a3) are expressed in the brain, and two (a1, a3) predominantly in neurons. The a3 isoform, encoded by ATP1A3, is critical to neuronal physiology, and a growing spectrum of neurological diseases are associated with ATP1A3 pathogenic variants, with ages of onset ranging from early childhood to adulthood. Here, we describe ATP1A3 variants encoding dysfunctional a3 subunits in children affected by polymicrogyria, a developmental malformation of the cerebral cortex characterized by abnormal folding and laminar organization. To gain cell-biological insights into the spatiotemporal dynamics of prenatal ATP1A3 expression, we established a transcriptional atlas of ATP1A3 expression during cortical development using mRNA in situ hybridization and transcriptomic profiling of ~125,000 individual cells with single-cell RNA sequencing (Drop-Seq) from various areas of the midgestational human neocortex. We find that fetal expression of ATP1A3 is restricted to a subset of excitatory neurons carrying transcriptional signatures of neuronal activity and maturation characteristic of the developing subplate. Furthermore, by performing Drop-Seq on ~52,000 nuclei from four different areas of an infant human neocortex, we show that ATP1A3 expression persists throughout early postnatal development, not only within excitatory neurons across all cortical layers, but also and more predominantly in inhibitory neurons, with specific enrichment in fast-spiking basket cells. In addition, we show that ATP1A3 expression, both in fetal and postnatal neurons, tends to be higher in frontal cortical areas than in occipital areas, in a pattern consistent with the rostro-caudal maturation gradient of the human neocortex. Finally, we discover distinct co-expression patterns linking catalytic  subunit isoforms (ATP1A1,2,3) and auxiliary isoforms (ATP1B1,2,3), suggesting the ATPase pump may form non-redundant, cell-type specific -{beta} combinations. Together, the importance of the developmental phenotypes and dynamic expression patterns of ATP1A3 point to a key role for a3 in the development and function of human cortex.
]]></description>
<dc:creator>Smith, R. S.</dc:creator>
<dc:creator>Florio, M.</dc:creator>
<dc:creator>Akula, S.</dc:creator>
<dc:creator>Neil, J.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Hill, R. S.</dc:creator>
<dc:creator>Goldman, M.</dc:creator>
<dc:creator>Mullally, C.</dc:creator>
<dc:creator>Reed, N.</dc:creator>
<dc:creator>Bello-Espinosa, L.</dc:creator>
<dc:creator>Sarnat, L. F.</dc:creator>
<dc:creator>Monteiro, F.</dc:creator>
<dc:creator>Erasmo, C.</dc:creator>
<dc:creator>Pinto e Vairo, F.</dc:creator>
<dc:creator>Morava, E.</dc:creator>
<dc:creator>Barkovich, A. J.</dc:creator>
<dc:creator>Gonzalez-Heydrich, J.</dc:creator>
<dc:creator>Brownstein, C.</dc:creator>
<dc:creator>McCarroll, S.</dc:creator>
<dc:creator>Walsh, C. A.</dc:creator>
<dc:date>2020-10-04</dc:date>
<dc:identifier>doi:10.1101/2020.10.03.319137</dc:identifier>
<dc:title><![CDATA[Role for the Na+/K+ ATPase α 3 subunit in folding and lamination of the human neocortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.06.329011v1?rss=1">
<title>
<![CDATA[
De novo structural mutation rates and gamete-of-origin biases revealed through genome sequencing of 2,396 families 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.06.329011v1?rss=1</link>
<description><![CDATA[
Each human genome includes de novo mutations that arose during gametogenesis. While these germline mutations represent a fundamental source of new genetic diversity, they can also create deleterious alleles that impact fitness. The germline mutation rate for single nucleotide variants and factors that significantly influence this rate, such as parental age, are now well established. However, far less is known about the frequency, distribution, and features that impact de novo structural mutations. We report a large, family-based study of germline mutations, excluding aneuploidy, that affect genome structure among 572 genomes from 33 families in a multigenerational CEPH-Utah cohort and 2,363 cases of non-familial autism spectrum disorder (ASD), 1,938 unaffected siblings, and both parents (9,599 genomes in total). We find that de novo structural mutations detected by alignment-based, short-read WGS occurred at an overall rate of at least 0.160 events per genome in unaffected individuals and was significantly higher (0.206 per genome) in ASD cases. In both probands and unaffected samples, nearly 73% of de novo structural mutations arose in paternal gametes, and predict most de novo structural mutations to be caused by mutational mechanisms that do not require sequence homology. After multiple testing correction we did not observe a statistically significant correlation between parental age and the rate of de novo structural variation in offspring. These results highlight that a spectrum of mutational mechanisms contribute to germline structural mutations, and that these mechanisms likely have markedly different rates and selective pressures than those leading to point mutations.
]]></description>
<dc:creator>Belyeu, J. R.</dc:creator>
<dc:creator>Brand, H.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Pedersen, B. S.</dc:creator>
<dc:creator>Feusier, J.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Nicholas, T. J.</dc:creator>
<dc:creator>Baird, L.</dc:creator>
<dc:creator>Devlin, B.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Jorde, L. B.</dc:creator>
<dc:creator>Talkowski, M. E.</dc:creator>
<dc:creator>Quinlan, A. R.</dc:creator>
<dc:date>2020-10-08</dc:date>
<dc:identifier>doi:10.1101/2020.10.06.329011</dc:identifier>
<dc:title><![CDATA[De novo structural mutation rates and gamete-of-origin biases revealed through genome sequencing of 2,396 families]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.07.321083v1?rss=1">
<title>
<![CDATA[
Tractography dissection variability: what happens when 42 groups dissect 14 white matter bundles on the same dataset? 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.07.321083v1?rss=1</link>
<description><![CDATA[
White matter bundle segmentation using diffusion MRI fiber tractography has become the method of choice to identify white matter fiber pathways in vivo in human brains. However, like other analyses of complex data, there is considerable variability in segmentation protocols and techniques. This can result in different reconstructions of the same intended white matter pathways, which directly affects tractography results, quantification, and interpretation. In this study, we aim to evaluate and quantify the variability that arises from different protocols for bundle segmentation. Through an open call to users of fiber tractography, including anatomists, clinicians, and algorithm developers, 42 independent teams were given processed sets of human whole-brain streamlines and asked to segment 14 white matter fascicles on six subjects. In total, we received 57 different bundle segmentation protocols, which enabled detailed volume-based and streamline-based analyses of agreement and disagreement among protocols for each fiber pathway. Results show that even when given the exact same sets of underlying streamlines, the variability across protocols for bundle segmentation is greater than all other sources of variability in the virtual dissection process, including variability within protocols and variability across subjects. In order to foster the use of tractography bundle dissection in routine clinical settings, and as a fundamental analytical tool, future endeavors must aim to resolve and reduce this heterogeneity. Although external validation is needed to verify the anatomical accuracy of bundle dissections, reducing heterogeneity is a step towards reproducible research and may be achieved through the use of standard nomenclature and definitions of white matter bundles and well-chosen constraints and decisions in the dissection process.
]]></description>
<dc:creator>Schilling, K. G.</dc:creator>
<dc:creator>Rheault, F.</dc:creator>
<dc:creator>Petit, L.</dc:creator>
<dc:creator>Hansen, C. B.</dc:creator>
<dc:creator>Nath, V.</dc:creator>
<dc:creator>Yeh, F.-C.</dc:creator>
<dc:creator>Girard, G.</dc:creator>
<dc:creator>Barakovic, M.</dc:creator>
<dc:creator>Rafael-Patino, J.</dc:creator>
<dc:creator>Yu, T.</dc:creator>
<dc:creator>Fischi-Gomez, E.</dc:creator>
<dc:creator>Pizzolato, M.</dc:creator>
<dc:creator>Ocampo-Pineda, M.</dc:creator>
<dc:creator>Schiavi, S.</dc:creator>
<dc:creator>Canales-Rodriguez, E. J.</dc:creator>
<dc:creator>Daducci, A.</dc:creator>
<dc:creator>Granziera, C.</dc:creator>
<dc:creator>Innocenti, G.</dc:creator>
<dc:creator>Thiran, J.-P.</dc:creator>
<dc:creator>Mancini, L.</dc:creator>
<dc:creator>Wastling, S.</dc:creator>
<dc:creator>Cocozza, S.</dc:creator>
<dc:creator>Petracca, M.</dc:creator>
<dc:creator>Pontillo, G.</dc:creator>
<dc:creator>Mancini, M.</dc:creator>
<dc:creator>Vos, S. B.</dc:creator>
<dc:creator>Vakharia, V. N.</dc:creator>
<dc:creator>Duncan, J. S.</dc:creator>
<dc:creator>Melero, H.</dc:creator>
<dc:creator>Manzanedo, L.</dc:creator>
<dc:creator>Sanz-Morales, E.</dc:creator>
<dc:creator>Pena-Melian, A.</dc:creator>
<dc:creator>Calamante, F.</dc:creator>
<dc:creator>Attye, A.</dc:creator>
<dc:creator>Cabeen, R. P.</dc:creator>
<dc:creator>Korobova, L.</dc:creator>
<dc:creator>Toga, A. W.</dc:creator>
<dc:creator>Ambili Vijayakumari, A.</dc:creator>
<dc:creator>Parker, D.</dc:creator>
<dc:creator>Verma, R.</dc:creator>
<dc:creator>Radwan, A.</dc:creator>
<dc:creator>Sun</dc:creator>
<dc:date>2020-10-08</dc:date>
<dc:identifier>doi:10.1101/2020.10.07.321083</dc:identifier>
<dc:title><![CDATA[Tractography dissection variability: what happens when 42 groups dissect 14 white matter bundles on the same dataset?]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.07.329698v1?rss=1">
<title>
<![CDATA[
Neural dynamics of semantic categorization in semantic variant of Primary Progressive Aphasia. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.07.329698v1?rss=1</link>
<description><![CDATA[
Semantic representations are processed along a posterior-to-anterior gradient reflecting a shift from perceptual (e.g., it has eight legs) to conceptual (e.g., venomous spiders are rare) information. One critical region is the anterior temporal lobe (ATL): patients with semantic variant primary progressive aphasia (svPPA), a clinical syndrome associated with ATL neurodegeneration, manifest a deep loss of semantic knowledge. We test the hypothesis that svPPA patients perform semantic tasks by over-recruiting areas implicated in perceptual processing. We compared MEG recordings of svPPA patients and healthy controls during a categorization task. While behavioral performance did not differ, svPPA patients showed indications of greater activation over bilateral occipital cortices and superior temporal gyrus, and inconsistent engagement of frontal regions. These findings suggest a pervasive reorganization of brain networks in response to ATL neurodegeneration: the loss of this critical hub leads to a dysregulated (semantic) control system, and defective semantic representations are seemingly compensated via enhanced perceptual processing.

Impact StatementFollowing anterior temporal lobe neurodegeneration, defective semantic representations are compensated via enhanced perceptual processing and associated with a dysregulation of the semantic control system.
]]></description>
<dc:creator>Borghesani, V.</dc:creator>
<dc:creator>Dale, C. L.</dc:creator>
<dc:creator>Lukic, S.</dc:creator>
<dc:creator>Hinkley, L. B. N.</dc:creator>
<dc:creator>Lauricella, M.</dc:creator>
<dc:creator>Shwe, W.</dc:creator>
<dc:creator>Miziuri, D.</dc:creator>
<dc:creator>Honma, S.</dc:creator>
<dc:creator>Miller, Z.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Houde, J.</dc:creator>
<dc:creator>Gorno-Tempini, M. L.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2020-10-09</dc:date>
<dc:identifier>doi:10.1101/2020.10.07.329698</dc:identifier>
<dc:title><![CDATA[Neural dynamics of semantic categorization in semantic variant of Primary Progressive Aphasia.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.09.332676v1?rss=1">
<title>
<![CDATA[
Cytonemes with complex geometries and composition extend into invaginations of target cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.09.332676v1?rss=1</link>
<description><![CDATA[
Cytonemes are specialized filopodia that mediate paracrine signaling in Drosophila and other animals. Studies using fluorescence confocal microscopy (CM) established their general paths, cell targets, and essential roles in signaling. To investigate details unresolvable by CM, we used high pressure freezing and electron microscopy to visualize cytoneme structures, paths, contents, and contacts. We observed cytonemes previously seen by CM in the Drosophila wing imaginal disc system, including in disc, tracheal air sac primordium (ASP), and myoblast cytonemes, and identified cytonemes extending into invaginations of target cells and cytonemes connecting ASP cells and connecting myoblasts. Diameters of cytoneme shafts vary between wide (206 {+/-} 51.8 nm) and thin (55.9 {+/-}16.2 nm) segments at regular intervals. Actin, ribosomes, and membranous compartments are present throughout; rough endoplasmic reticulum and mitochondria are in wider proximal sections. These results reveal novel structural features of filopodia and provide a basis for understanding cytoneme cell biology and function.

SummaryCytoneme signaling filopodia of Drosophila cells have regions defined by oscillating diameters, and contents that include ER, mitochondria and ribosomes, and form contacts along invaginations of target cell membranes.
]]></description>
<dc:creator>Kornberg, T. B.</dc:creator>
<dc:creator>Wood, B.</dc:creator>
<dc:creator>Baena, V.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Terasaki, M.</dc:creator>
<dc:date>2020-10-09</dc:date>
<dc:identifier>doi:10.1101/2020.10.09.332676</dc:identifier>
<dc:title><![CDATA[Cytonemes with complex geometries and composition extend into invaginations of target cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.09.334078v1?rss=1">
<title>
<![CDATA[
CoRa -A general approach for quantifying biological feedback control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.09.334078v1?rss=1</link>
<description><![CDATA[
Feedback control is a fundamental underpinning of life, underlying homeostasis of biological processes at every scale of organization, from cells to ecosystems. The ability to evaluate the contribution and limitations of feedback control mechanisms operating in cells is a critical step for understanding and ultimately designing feedback control systems with biological molecules. Here, we introduce CoRa -or Control Ratio-, a general framework that quantifies the contribution of a biological feedback control mechanism to adaptation using a mathematically controlled comparison to an identical system that does not contain the feedback. CoRa provides a simple and intuitive metric with broad applicability to biological feedback systems.
]]></description>
<dc:creator>Gomez-Schiavon, M.</dc:creator>
<dc:creator>El-Samad, H.</dc:creator>
<dc:date>2020-10-10</dc:date>
<dc:identifier>doi:10.1101/2020.10.09.334078</dc:identifier>
<dc:title><![CDATA[CoRa -A general approach for quantifying biological feedback control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.12.336784v1?rss=1">
<title>
<![CDATA[
Single cell transcriptome analysis defines novel heterogeneity within the pancreatic ductal tree 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.12.336784v1?rss=1</link>
<description><![CDATA[
Lineage tracing using genetically engineered mouse models is an essential tool for investigating cell-fate decisions of progenitor cells and biology of mature cell types, with relevance to physiology and disease progression. To study disease development, an inventory of an organs cell types and understanding of physiologic function is paramount. Here, we performed single-cell RNA sequencing to examine heterogeneity of murine pancreatic duct cells, pancreatobiliary cells, and intrapancreatic bile duct cells. We describe an epithelial-mesenchymal transitory axis in our three pancreatic duct subpopulations and identify SPP1 as a regulator of this fate decision as well as human duct cell de-differentiation. Our results further identify functional heterogeneity within pancreatic duct subpopulations by elucidating a role for Geminin in accumulation of DNA damage in the setting of chronic pancreatitis. Our findings implicate diverse functional roles for subpopulations of pancreatic duct cells in maintenance of duct cell identity and disease progression and establish a comprehensive road map of murine pancreatic duct cell, pancreatobiliary cell, and intrapancreatic bile duct cell homeostasis.

SIGNIFICANCEMurine models are extensively used for pancreatic lineage tracing experiments and investigation of pancreatic disease progression. Here, we describe the transcriptome of murine pancreatic duct cells, intrapancreatic bile duct cells, and pancreatobiliary cells at single cell resolution. Our analysis defines novel heterogeneity within the pancreatic ductal tree and supports the paradigm that more than one population of pancreatic duct cells harbors progenitor capacity. We identify and validate unique functional properties of subpopulations of pancreatic duct cells including an epithelial-mesenchymal transcriptomic axis and roles in chronic pancreatic inflammation.
]]></description>
<dc:creator>Hendley, A. M.</dc:creator>
<dc:creator>Rao, A. A.</dc:creator>
<dc:creator>Leonhardt, L.</dc:creator>
<dc:creator>Ashe, S.</dc:creator>
<dc:creator>Smith, J. A.</dc:creator>
<dc:creator>Giacometti, S.</dc:creator>
<dc:creator>Peng, X. L.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Berrios, D.</dc:creator>
<dc:creator>Pawlak, M.</dc:creator>
<dc:creator>Li, L. Y.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Collisson, E. A.</dc:creator>
<dc:creator>Anderson, M.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Yeh, J. J.</dc:creator>
<dc:creator>Chun, J. Y.</dc:creator>
<dc:creator>Kim, G. E.</dc:creator>
<dc:creator>Weaver, V. M.</dc:creator>
<dc:creator>Hebrok, M.</dc:creator>
<dc:date>2020-10-12</dc:date>
<dc:identifier>doi:10.1101/2020.10.12.336784</dc:identifier>
<dc:title><![CDATA[Single cell transcriptome analysis defines novel heterogeneity within the pancreatic ductal tree]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.13.337865v1?rss=1">
<title>
<![CDATA[
On the role of photoreceptor identity in controlling accurate wiring of the Drosophila visual circuit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.13.337865v1?rss=1</link>
<description><![CDATA[
During development, neurons extend in search of synaptic partners. Precise control of axon extension velocity can therefore be crucial to ensuring proper circuit formation. How velocity is regulated - particularly by the extending axons themselves - remains poorly understood. Here, we investigate this question in the Drosophila visual system, where photoreceptors make precise connections with a specific set of synaptic partners that together create a circuit underpinning neural superposition (NSP). We used a combination of genetic perturbations and quantitative image analysis to investigate the influence of cell identity on growth cone velocity and subsequent spatial-temporal coincidence of presynaptic and postsynaptic neurons. Our study provides a case study of how cell autonomous properties of presynaptic axons play a pivotal role in controlling the dynamics of growing axons and determining the formation of a precise neuronal circuit.
]]></description>
<dc:creator>Altschuler, S. J.</dc:creator>
<dc:creator>Ji, W.</dc:creator>
<dc:creator>Wu, L. F.</dc:creator>
<dc:date>2020-10-13</dc:date>
<dc:identifier>doi:10.1101/2020.10.13.337865</dc:identifier>
<dc:title><![CDATA[On the role of photoreceptor identity in controlling accurate wiring of the Drosophila visual circuit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.22.348631v1?rss=1">
<title>
<![CDATA[
PTBP1 Regulates Injury Responses and Sensory Pathways in Adult Peripheral Neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.22.348631v1?rss=1</link>
<description><![CDATA[
Polypyrimidine Tract Binding Protein 1 (PTBP1) is expressed only at embryonic stages in central neurons. Its downregulation triggers neuronal differentiation in precursor and non-neuronal cells, an approach recently used to generate neurons de novo for amelioration of neurodegenerative disorders. Moreover, PTBP1 is replaced by its paralog PTBP2 in mature central neurons. Surprisingly, we found both proteins co-expressed in adult sensory and motor neurons, with PTBP2 restricted mainly to the nucleus, while PTBP1 shows strong axonal localization. Levels of axonal PTBP1 increased markedly after peripheral nerve injury, and its cargos include mRNAs involved in axonal growth and regeneration, such as importin {beta}1 and RhoA. Perturbation of PTBP1 affects neuronal injury responses, axon outgrowth and sensation in vivo. Thus, PTBP1 has roles in sensory function and regenerative capacity of adult sensory neurons. These findings suggest that caution may be required before considering targeting PTBP1 for therapeutic purposes.
]]></description>
<dc:creator>Alber, S.</dc:creator>
<dc:creator>Di-Matteo, P.</dc:creator>
<dc:creator>Zdradzinski, M. D.</dc:creator>
<dc:creator>Marvaldi, L.</dc:creator>
<dc:creator>Kawaguchi, R.</dc:creator>
<dc:creator>Medzihradszky, K. F.</dc:creator>
<dc:creator>Doron-Mandel, E.</dc:creator>
<dc:creator>Okladnikov, N.</dc:creator>
<dc:creator>Rishal, I.</dc:creator>
<dc:creator>Nevo, R.</dc:creator>
<dc:creator>Lee, S. J.</dc:creator>
<dc:creator>Sahoo, P. K.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Coppola, G.</dc:creator>
<dc:creator>Twiss, J. L.</dc:creator>
<dc:creator>Fainzilber, M.</dc:creator>
<dc:date>2020-10-22</dc:date>
<dc:identifier>doi:10.1101/2020.10.22.348631</dc:identifier>
<dc:title><![CDATA[PTBP1 Regulates Injury Responses and Sensory Pathways in Adult Peripheral Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.23.352666v1?rss=1">
<title>
<![CDATA[
Transcriptomics-based drug repositioning pipeline identifies therapeutic candidates for COVID-19 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.23.352666v1?rss=1</link>
<description><![CDATA[
The novel SARS-CoV-2 virus emerged in December 2019 and has few effective treatments. We applied a computational drug repositioning pipeline to SARS-CoV-2 differential gene expression signatures derived from publicly available data. We utilized three independent published studies to acquire or generate lists of differentially expressed genes between control and SARS-CoV-2-infected samples. Using a rank-based pattern matching strategy based on the Kolmogorov-Smirnov Statistic, the signatures were queried against drug profiles from Connectivity Map (CMap). We validated sixteen of our top predicted hits in live SARS-CoV-2 antiviral assays in either Calu-3 or 293T-ACE2 cells. Validation experiments in human cell lines showed that 11 of the 16 compounds tested to date (including clofazimine, haloperidol and others) had measurable antiviral activity against SARS-CoV-2. These initial results are encouraging as we continue to work towards a further analysis of these predicted drugs as potential therapeutics for the treatment of COVID-19.
]]></description>
<dc:creator>Le, B. L.</dc:creator>
<dc:creator>Andreoletti, G.</dc:creator>
<dc:creator>Oskotsky, T.</dc:creator>
<dc:creator>Vallejo-Gracia, A.</dc:creator>
<dc:creator>Rosales, R.</dc:creator>
<dc:creator>Yu, K. Z.</dc:creator>
<dc:creator>Kosti, I.</dc:creator>
<dc:creator>Leon, K. E.</dc:creator>
<dc:creator>Bunis, D. G.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Kumar, G. R.</dc:creator>
<dc:creator>White, K. M.</dc:creator>
<dc:creator>Garcia-Sastre, A. E.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2020-10-23</dc:date>
<dc:identifier>doi:10.1101/2020.10.23.352666</dc:identifier>
<dc:title><![CDATA[Transcriptomics-based drug repositioning pipeline identifies therapeutic candidates for COVID-19]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.27.352716v1?rss=1">
<title>
<![CDATA[
Cytoplasm Biophysical Properties Madolute Microtubule Dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.27.352716v1?rss=1</link>
<description><![CDATA[
The cytoplasm is a crowded, visco-elastic environment whose physical properties change according to physiological or developmental states. How the physical properties of the cytoplasm impact cellular functions in vivo remain poorly understood. Here, we probed the effects of cytoplasmic concentration on microtubules by applying osmotic shifts to fission yeast, moss, and mammalian cells. We show that both the rates of microtubule polymerization and depolymerization scale linearly and inversely with cytoplasmic concentration; an increase in cytoplasmic concentration decreases the rates of microtubule polymerization and depolymerization proportionally, while a decrease in cytoplasmic concentration leads to the opposite. Numerous lines of evidence indicate that these effects are due to changes in cytoplasmic viscosity rather than cellular stress responses or macromolecular crowding per se. We reconstituted these effects on microtubules in vitro by tuning viscosity. Our findings indicate that, even in normal conditions, the viscosity of cytoplasm modulates the reactions underlying microtubule dynamic behaviors.
]]></description>
<dc:creator>Molines, A. T.</dc:creator>
<dc:creator>Lemiere, J.</dc:creator>
<dc:creator>Edrington, C. H.</dc:creator>
<dc:creator>Hsu, C.-T.</dc:creator>
<dc:creator>Steinmark, I. E.</dc:creator>
<dc:creator>Suhling, K.</dc:creator>
<dc:creator>Goshima, G.</dc:creator>
<dc:creator>Holt, L. J.</dc:creator>
<dc:creator>Brouhard, G.</dc:creator>
<dc:creator>Chang, F. T.</dc:creator>
<dc:date>2020-10-28</dc:date>
<dc:identifier>doi:10.1101/2020.10.27.352716</dc:identifier>
<dc:title><![CDATA[Cytoplasm Biophysical Properties Madolute Microtubule Dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.28.359042v1?rss=1">
<title>
<![CDATA[
Ethacridine inhibits SARS-CoV-2 by inactivating viral particles in cellular models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.28.359042v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 is the coronavirus that causes the respiratory disease COVID-19, which is now the third-leading cause of death in the United States. The FDA has recently approved remdesivir, an inhibitor of SARS-CoV-2 replication, to treat COVID-19, though recent data from the WHO shows little to no benefit with use of this anti-viral agent. Here we report the discovery of ethacridine, a safe antiseptic use in humans, as a potent drug for use against SARS-CoV-2 (EC50 ~ 0.08 M). Ethacridine was identified via high-throughput screening of an FDA-approved drug library in living cells using a fluorescent assay. Interestingly, the main mode of action of ethacridine is through inactivation of viral particles, preventing their binding to the host cells. Indeed, ethacridine is effective in various cell types, including primary human nasal epithelial cells. Taken together, these data identify a promising, potent, and new use of the old drug possessing a distinct mode of action for inhibiting SARS-CoV-2.
]]></description>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Lidsky, P.</dc:creator>
<dc:creator>Xiao, Y.</dc:creator>
<dc:creator>Wu, C.-T.</dc:creator>
<dc:creator>Garcia-Knight, M.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Nakayama, T.</dc:creator>
<dc:creator>Nayak, J. V.</dc:creator>
<dc:creator>Jackson, P. K.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:creator>Shu, X.</dc:creator>
<dc:date>2020-10-28</dc:date>
<dc:identifier>doi:10.1101/2020.10.28.359042</dc:identifier>
<dc:title><![CDATA[Ethacridine inhibits SARS-CoV-2 by inactivating viral particles in cellular models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.28.359935v1?rss=1">
<title>
<![CDATA[
Global Absence and Targeting of Protective Immune States in Severe COVID-19. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.28.359935v1?rss=1</link>
<description><![CDATA[
While SARS-CoV-2 infection has pleiotropic and systemic effects in some patients, many others experience milder symptoms. We sought a holistic understanding of the severe/mild distinction in COVID-19 pathology, and its origins. We performed a whole-blood preserving single-cell analysis protocol to integrate contributions from all major cell types including neutrophils, monocytes, platelets, lymphocytes and the contents of serum. Patients with mild COVID-19 disease display a coordinated pattern of interferon-stimulated gene (ISG) expression across every cell population and these cells are systemically absent in patients with severe disease. Severe COVID-19 patients also paradoxically produce very high anti-SARS-CoV-2 antibody titers and have lower viral load as compared to mild disease. Examination of the serum from severe patients demonstrates that they uniquely produce antibodies with multiple patterns of specificity against interferon-stimulated cells and that those antibodies functionally block the production of the mild disease-associated ISG-expressing cells. Overzealous and auto-directed antibody responses pit the immune system against itself in many COVID-19 patients and this defines targets for immunotherapies to allow immune systems to provide viral defense.

One Sentence SummaryIn severe COVID-19 patients, the immune system fails to generate cells that define mild disease; antibodies in their serum actively prevents the successful production of those cells.
]]></description>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Courau, T.</dc:creator>
<dc:creator>Kuhn, N. F.</dc:creator>
<dc:creator>Hu, K. H.</dc:creator>
<dc:creator>Ray, A.</dc:creator>
<dc:creator>Chen, W. S.</dc:creator>
<dc:creator>Clearly, S. J.</dc:creator>
<dc:creator>Chew, N. W.</dc:creator>
<dc:creator>Kushnoor, D.</dc:creator>
<dc:creator>Reeder, G. C.</dc:creator>
<dc:creator>Shen, A.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Hiam-Galvez, K. J.</dc:creator>
<dc:creator>Munoz-Sandoval, P.</dc:creator>
<dc:creator>Zhu, W. S.</dc:creator>
<dc:creator>Lee, D. S.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>You, R.</dc:creator>
<dc:creator>Magnen, M.</dc:creator>
<dc:creator>Rodriguez, L.</dc:creator>
<dc:creator>Leligdowicz, A.</dc:creator>
<dc:creator>Zamecnik, C. R.</dc:creator>
<dc:creator>Loudermilk, R. P.</dc:creator>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Looney, M. R.</dc:creator>
<dc:creator>Chan, V.</dc:creator>
<dc:creator>Ward, A.</dc:creator>
<dc:creator>Carrillo, S.</dc:creator>
<dc:creator>Matthay, M.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:creator>Woodruff, P. G.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>Kangelaris, K.</dc:creator>
<dc:creator>Hendrickson, C. M.</dc:creator>
<dc:creator>Calfee, C.</dc:creator>
<dc:creator>Rao, A. A.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2020-10-29</dc:date>
<dc:identifier>doi:10.1101/2020.10.28.359935</dc:identifier>
<dc:title><![CDATA[Global Absence and Targeting of Protective Immune States in Severe COVID-19.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.30.362715v1?rss=1">
<title>
<![CDATA[
Ethnicity-specific transcriptomic variation in immune cells and correlation with disease activity in systemic lupus erythematosus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.30.362715v1?rss=1</link>
<description><![CDATA[
Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease in which outcomes vary among different racial groups. The aim of this study is to leverage large-scale transcriptomic data from diverse populations to better sub-classify SLE patients into more clinically actionable groups. We leverage cell sorted RNA-seq data (CD14+ monocytes, B cells, CD4+T cells, and NK cells) from 120 SLE patients (63 Asian and 57 White individuals) and apply a four tier analytical approach to identify SLE subgroups within this multiethnic cohort: unsupervised clustering, differential expression analyses, gene co-expression analyses, and machine learning. K-means clustering on the individual cell type data resulted in three clusters for CD4 and CD14, and two clusters for B cells and NK cells. Correlation analysis revealed significant positive associations between the transcriptomic clusters of each immune cell and clinical parameters including disease activity and ethnicity. We then explored differentially expressed genes between Asian and White groups for each cell-type. The shared differentially expressed genes across the four cell types were involved in SLE or other autoimmune related pathways. Co-expression analysis identified similarly regulated genes across samples and grouped these genes into modules. Samples were grouped into White-high, Asians-high (high disease activity defined by SLEDAI score >=6) and White-low, Asians-low (SLEDAI < 6). Random forest classification of disease activity in the White and Asian cohorts showed the best classification in CD4+ T cells in White. The results from these analyses will help stratify patients based on their gene expression signatures to enable precision medicine for SLE.
]]></description>
<dc:creator>Andreoletti, G.</dc:creator>
<dc:creator>Lanata, C. M.</dc:creator>
<dc:creator>Paranjpe, I.</dc:creator>
<dc:creator>Jain, T. S.</dc:creator>
<dc:creator>Nititham, J.</dc:creator>
<dc:creator>Taylor, K. E.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Maliskova, L.</dc:creator>
<dc:creator>Jimmie Ye, C.</dc:creator>
<dc:creator>Katz, P.</dc:creator>
<dc:creator>Dall Era, M.</dc:creator>
<dc:creator>Yazdany, J.</dc:creator>
<dc:creator>Criswell, L. A.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2020-11-01</dc:date>
<dc:identifier>doi:10.1101/2020.10.30.362715</dc:identifier>
<dc:title><![CDATA[Ethnicity-specific transcriptomic variation in immune cells and correlation with disease activity in systemic lupus erythematosus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.03.366914v1?rss=1">
<title>
<![CDATA[
Transcriptional Network Orchestrating Regional Patterning of Cortical Progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.03.366914v1?rss=1</link>
<description><![CDATA[
We uncovered a transcription factor (TF) network that regulates cortical regional patterning. Screening the expression of hundreds of TFs in the developing mouse cortex identified 38 TFs that are expressed in gradients in the ventricular zone (VZ). We tested whether their cortical expression was altered in mutant mice with known patterning defects (Emx2, Nr2f1 and Pax6), which enabled us to define a cortical regionalization TF network (CRTFN). To identify genomic programming underlying this network, we performed TF ChIP-seq and chromatin-looping conformation to identify enhancer-gene interactions. To map enhancers involved in regional patterning of cortical progenitors, we performed assays for epigenomic marks and DNA accessibility in VZ cells purified from wild-type and patterning mutant mice. This integrated approach has identified a CRTFN and VZ enhancers involved in cortical regional patterning.
]]></description>
<dc:creator>Ypsilanti, A. R.</dc:creator>
<dc:creator>Pattabiraman, K.</dc:creator>
<dc:creator>Catta-Preta, R.</dc:creator>
<dc:creator>Golonzhka, O.</dc:creator>
<dc:creator>Lindtner, S.</dc:creator>
<dc:creator>Tang, K.</dc:creator>
<dc:creator>Jones, I.</dc:creator>
<dc:creator>Abnousi, A.</dc:creator>
<dc:creator>Juric, I.</dc:creator>
<dc:creator>Hu, M.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Dickel, D. E.</dc:creator>
<dc:creator>Visel, A.</dc:creator>
<dc:creator>Pennachio, L. A.</dc:creator>
<dc:creator>Hawrylycz, M.</dc:creator>
<dc:creator>Thompson, C.</dc:creator>
<dc:creator>Zeng, H.</dc:creator>
<dc:creator>Barozzi, I.</dc:creator>
<dc:creator>Nord, A. S.</dc:creator>
<dc:creator>Rubenstein, J. L. R.</dc:creator>
<dc:date>2020-11-04</dc:date>
<dc:identifier>doi:10.1101/2020.11.03.366914</dc:identifier>
<dc:title><![CDATA[Transcriptional Network Orchestrating Regional Patterning of Cortical Progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.05.370569v1?rss=1">
<title>
<![CDATA[
Activating Immune Recognition in Pancreatic Ductal Adenocarcinoma Using Autophagy Inhibition, MEK blockade and CD40 Agonism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.05.370569v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDA) patients have not yet benefitted from the revolution in cancer immunotherapy due in large part to the dominantly immunosuppressive tumor microenvironment (TME). MEK inhibition combined with autophagy inhibition leads to transient tumor responses in some PDA patients. We find that co-inhibition of MEK (using cobimetinib, COBI) and autophagy (using mefloquine, MFQ), but not either treatment alone, activates the Type I Interferon/STING pathway in tumor cells which in turn reprogram tumor associated macrophages (TAMs) in paracrine to foster an immunogenic switch. This effect is augmented by a CD40 agonist (aCD40). Triple therapy (COBI+MFQ+aCD40) achieved cytotoxic T cell activation in an immunologically "cold" mouse PDA model, leading to enhanced anti-tumor immunity. Collectively, MEK and autophagy co-inhibition coupled with CD40 agonism invokes immuno-reprograming and is an attractive therapeutic approach for PDA immunotherapy development.
]]></description>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Courau, T.</dc:creator>
<dc:creator>Lupin-Jimenez, L.</dc:creator>
<dc:creator>Borison, J.</dc:creator>
<dc:creator>Ritchie, A. J.</dc:creator>
<dc:creator>Mayer, A. T.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Collisson, E. A.</dc:creator>
<dc:date>2020-11-06</dc:date>
<dc:identifier>doi:10.1101/2020.11.05.370569</dc:identifier>
<dc:title><![CDATA[Activating Immune Recognition in Pancreatic Ductal Adenocarcinoma Using Autophagy Inhibition, MEK blockade and CD40 Agonism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.08.371922v1?rss=1">
<title>
<![CDATA[
TSC1 loss-of-function increases risk for tauopathy by inducing tau acetylation and preventing autophagy-mediated tau clearance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.08.371922v1?rss=1</link>
<description><![CDATA[
Age-associated neurodegenerative disorders demonstrating tau-laden intracellular inclusions, including Alzheimers disease (AD), frontotemporal lobar degeneration (FTLD) and progressive supranuclear palsy (PSP), are collectively known as tauopathies. The vast majority of human tauopathies accumulate non-mutant tau rather than mutant forms of the protein, yet cell and animal models for non-mutant tauopathies are lacking. We previously linked a monoallelic mutation in the TSC1 gene to tau accumulation and FTLD. Now, we have identified new variants in TSC1 that predisposed to other tauopathies such as AD and PSP. These new TSC1 risk variants significantly decreased the half-life of TSC1/hamartin in vitro. Cellular and murine models of TSC1 haploinsufficiency (TSC1+/-) accumulated tau protein that exhibited aberrant acetylation on six lysine residues. Tau acetylation hindered its lysosomal degradation via chaperone-mediated autophagy leading to neuronal tau accumulation. Enhanced tau acetylation in TSC1+/- models was achieved through both an increase in p300 acetyltransferase activity and a decrease in SIRT1 deacetylase levels. Pharmacological modulation of either enzyme restored tau levels. Together, these studies substantiate TSC1 as a novel tauopathy risk gene and advance TSC1 haploinsufficiency as a new genetic model for tauopathy. In addition, these results promote acetylated tau as a rational target for diagnostic and therapeutic modalities in multiple tauopathies.
]]></description>
<dc:creator>Alquezar, C.</dc:creator>
<dc:creator>Schoch, K. M.</dc:creator>
<dc:creator>Geier, E. G.</dc:creator>
<dc:creator>Ramos, E. M.</dc:creator>
<dc:creator>Scrivo, A.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Argouarch, A. R.</dc:creator>
<dc:creator>Mlynarski, E. E.</dc:creator>
<dc:creator>Dombroski, B.</dc:creator>
<dc:creator>Yokoyama, J. S.</dc:creator>
<dc:creator>Cuervo, A. M.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Schellenberg, G. D.</dc:creator>
<dc:creator>Miller, T. M.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Kao, A. W.</dc:creator>
<dc:date>2020-11-08</dc:date>
<dc:identifier>doi:10.1101/2020.11.08.371922</dc:identifier>
<dc:title><![CDATA[TSC1 loss-of-function increases risk for tauopathy by inducing tau acetylation and preventing autophagy-mediated tau clearance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.13.382234v1?rss=1">
<title>
<![CDATA[
The primary cilium is required for MC4R control of food intake and body weight 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.13.382234v1?rss=1</link>
<description><![CDATA[
The Melanocortin-4 Receptor (MC4R) plays a critical role in the long-term regulation of energy homeostasis and mutations in MC4R are the most common cause of monogenic obesity. However, the precise molecular and cellular mechanisms underlying the maintenance of energy balance within MC4R expressing neurons are unknown. We recently reported that MC4R localizes to primary cilia, a cellular organelle that allows for partitioning of incoming cellular signals, raising the question of whether MC4R functions there. Here, using mouse genetic approaches, we found that cilia are required specifically on MC4R-expressing neurons to restrain feeding behavior. Moreover, these cilia were critical for pharmacological activators of MC4R to exert an anorexigenic effect. MC4R is expressed in multiple brain regions. Using targeted deletion of primary cilia, we found that cilia in the paraventricular nucleus (PVN) of the hypothalamus are essential to restrict food intake. MC4R activation increases adenylyl cyclase activity. Like removing cilia, inhibiting adenylyl cyclase activity in the cilia of MC4R-expressing neurons of the PVN caused hyperphagia and obesity. Thus, MC4R signals via cilia of PVN neurons to control food intake and body weight. We propose that defects in ciliary localization of MC4R cause obesity in human inherited obesity syndromes and ciliopathies.
]]></description>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Bernard, A. A.</dc:creator>
<dc:creator>Comblain, F.</dc:creator>
<dc:creator>Yue, X.</dc:creator>
<dc:creator>Paillart, C.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Reiter, J.</dc:creator>
<dc:creator>Vaisse, C.</dc:creator>
<dc:date>2020-11-15</dc:date>
<dc:identifier>doi:10.1101/2020.11.13.382234</dc:identifier>
<dc:title><![CDATA[The primary cilium is required for MC4R control of food intake and body weight]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.13.382325v1?rss=1">
<title>
<![CDATA[
The single pass membrane protein MRAP2 regulates energy homeostasis by promoting primary cilia localization of the G protein-coupled receptor MC4R 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.13.382325v1?rss=1</link>
<description><![CDATA[
The G protein-coupled receptor MC4R (Melanocortin-4 Receptor) and its associated protein MRAP2 (Melanocortin Receptor-Associated Protein 2) are both essential for the regulation of food intake and body weight in humans and mice. MC4R localizes and functions at the neuronal primary cilium, a microtubule-based organelle that senses and relays extracellular signals. Here, we demonstrate that MRAP2 is critical for the ciliary localization and weight-regulating function of MC4R. Our data reveal that GPCR localization to primary cilia can require specific accessory proteins that may not be present in heterologous cell systems. Our findings also demonstrate the essential role of neuronal primary cilia localization of MC4R for adequate control of energy homeostasis and the obesity-promoting effect of genetic disruption of this pathway.
]]></description>
<dc:creator>Bernard, A. A.</dc:creator>
<dc:creator>Ojeda Naharros, I.</dc:creator>
<dc:creator>Bourgain-Guglielmetti, F.</dc:creator>
<dc:creator>Ciprin, J.</dc:creator>
<dc:creator>Yue, X.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>McDaid, E.</dc:creator>
<dc:creator>Nachury, M. V.</dc:creator>
<dc:creator>Reiter, J.</dc:creator>
<dc:creator>Vaisse, C.</dc:creator>
<dc:date>2020-11-15</dc:date>
<dc:identifier>doi:10.1101/2020.11.13.382325</dc:identifier>
<dc:title><![CDATA[The single pass membrane protein MRAP2 regulates energy homeostasis by promoting primary cilia localization of the G protein-coupled receptor MC4R]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.13.382374v1?rss=1">
<title>
<![CDATA[
Functional microstructure of CaV-mediated calcium signaling in the axon initial segment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.13.382374v1?rss=1</link>
<description><![CDATA[
The axon initial segment (AIS) is a specialized neuronal compartment in which synaptic input is converted into action potential output. This process is supported by a diverse complement of sodium, potassium, and calcium channels (CaV). Different classes of sodium and potassium channels are scaffolded at specific sites within the AIS, conferring unique functions, but how calcium channels are functionally distributed within the AIS is unclear. Here, we utilize conventional 2-photon laser scanning and diffraction-limited, high-speed spot 2-photon imaging to resolve action potential-evoked calcium dynamics in the AIS with high spatiotemporal resolution. In mouse layer 5 prefrontal pyramidal neurons, calcium influx was mediated by a mix of CaV2 and CaV3 channels that differentially localized to discrete regions. CaV3 functionally localized to produce nanodomain hotspots of calcium influx that coupled to ryanodine-dependent stores, whereas CaV2 localized to non-hotspot regions. Thus, different pools of CaVs appear to play distinct roles in AIS function.
]]></description>
<dc:creator>Lipkin, A. M.</dc:creator>
<dc:creator>Cunniff, M. M.</dc:creator>
<dc:creator>Spratt, P. W.</dc:creator>
<dc:creator>Lemke, S. M.</dc:creator>
<dc:creator>Bender, K. J.</dc:creator>
<dc:date>2020-11-13</dc:date>
<dc:identifier>doi:10.1101/2020.11.13.382374</dc:identifier>
<dc:title><![CDATA[Functional microstructure of CaV-mediated calcium signaling in the axon initial segment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.14.381749v1?rss=1">
<title>
<![CDATA[
GABAergic signaling promotes early-life seizures in epileptic SYNGAP1+/- mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.14.381749v1?rss=1</link>
<description><![CDATA[
ObjectiveSYNGAP1 encephalopathy is a developmental and epileptic encephalopathy caused by pathogenic loss of function variants. Syngap1-heterozygous (Het+/-) mice demonstrate progressive epilepsy with multiple seizure phenotypes in adulthood. Here, we investigate early-life seizures in Het+/- pups and explore of Syngap1 encephalopathy during development.

MethodsPost-natal day 7 (P7) and P12 mice were investigated by tethered video-electroencephalographic (vEEG). The effects of GABAergic drugs phenobarbital (PB) and pentylenetetrazol (PTZ) were investigated at P7 and P12, respectively. 24h tethered vEEG was performed at P24, and telemetric 24h vEEG with 6h sleep deprivation was performed at P35. The effect of perampanel (PMP), an AMPA receptor antagonist, was investigated at P24.

ResultsHet+/- mice have spontaneous early-life seizures that lack an overt behavioral phenotype. These subclinical seizures are refractory to PB, but the GABAA receptor (GABAAR) antagonist PTZ significantly reduced seizure frequency suggesting that GABAergic signaling may promote seizure generation in Het+/- pups. At juvenile ages, Het+/- pups recapitulated the early emergence of high gamma (35-50Hz) during NREM and disruption of behavioral-state gamma homeostasis. This biomarker was significantly exacerbated in Het+/- pups after increasing sleep pressure with sleep deprivation.

SignificanceSeizures during development have adverse effects on cognitive function. Therefore, an improved understanding of the SYNGAP1 epilepsy during developmental ages is necessary to delineate the deleterious interactions between aberrant synaptic function and recurrent seizures. The development of evidence-based therapies for early-life intervention will benefit from these insights.
]]></description>
<dc:creator>Sullivan, B. J.</dc:creator>
<dc:creator>Kipnis, P. A.</dc:creator>
<dc:creator>Ammanuel, S. G.</dc:creator>
<dc:creator>Kadam, S. D.</dc:creator>
<dc:date>2020-11-15</dc:date>
<dc:identifier>doi:10.1101/2020.11.14.381749</dc:identifier>
<dc:title><![CDATA[GABAergic signaling promotes early-life seizures in epileptic SYNGAP1+/- mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.17.387662v1?rss=1">
<title>
<![CDATA[
ATP hydrolysis coordinates the activities of two motors in a dimeric chromatin remodeling enzyme 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.17.387662v1?rss=1</link>
<description><![CDATA[
ATP-dependent chromatin remodelers are essential enzymes that restructure eukaryotic genomes to enable all DNA-based processes. The diversity and complexity of these processes are matched by the complexity of the enzymes that carry them out, making remodelers a challenging class of molecular motors to study by conventional methods. Here we use a single molecule biophysical assay to overcome some of these challenges, enabling a detailed mechanistic dissection of a paradigmatic remodeler reaction, that of sliding a nucleosome towards the longer DNA linker. We focus on how two motors of a dimeric remodeler coordinate to accomplish such directional sliding. We find that ATP hydrolysis by both motors promotes coordination, suggesting a role for ATP in resolving the competition for directional commitment. Furthermore, we show an artificially constitutive dimer is no more or less coordinated, but is more processive, suggesting a cell could modulate a remodelers oligomeric state to modulate local chromatin dynamics.
]]></description>
<dc:creator>Johnson, S. L.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:date>2020-11-18</dc:date>
<dc:identifier>doi:10.1101/2020.11.17.387662</dc:identifier>
<dc:title><![CDATA[ATP hydrolysis coordinates the activities of two motors in a dimeric chromatin remodeling enzyme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.23.394445v1?rss=1">
<title>
<![CDATA[
Regulation of Dopamine D1 Receptor Signaling at the Golgi Apparatus by a Low Affinity Dopamine Transporter 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.23.394445v1?rss=1</link>
<description><![CDATA[
Dopamine is the main catecholamine in the brain and the kidney, where it controls a number of physiological functions such as locomotion, cognition, emotion, endocrine regulation and renal function. As a membrane impermeant hormone and neurotransmitter, dopamine is thought to signal by binding and activating dopamine receptors, members of the G protein couple receptor (GPCR) family, only on the plasma membrane. Here, using novel nanobody-based biosensors, we demonstrate for the first time that the dopamine D1 receptor (D1DR), the primary mediator of dopaminergic signaling in the brain and kidney, not only functions on the plasma membrane but becomes activated at the Golgi apparatus in the presence of its ligand. We present evidence that activation of the Golgi pool of D1DR is dependent on Organic Cation Transporter 2 (OCT2), a dopamine transporter, providing an explanation for how the membrane impermeant dopamine accesses subcellular pools of D1DR. We further demonstrate that dopamine activates Golgi-D1DR in the striatal medium spiny neurons (MSN) and this activity depends on OCT2 function. We also introduce a new approach to selectively interrogate compartmentalized D1DR signaling by inhibiting Gs coupling, using a nanobody-based chemical recruitment system. Using this strategy, we show that Golgi-localized D1DRs regulate cAMP production and mediate local protein kinase A activation. Together, our data suggest that spatially compartmentalized signaling hubs are previously unappreciated regulatory aspects of D1DR signaling. Our data also provide further evidence for the role of transporters in regulating subcellular GPCR activity.
]]></description>
<dc:creator>Puri, N. M.</dc:creator>
<dc:creator>Romano, G. R.</dc:creator>
<dc:creator>Mai, Q. N.</dc:creator>
<dc:creator>Irannejad, R.</dc:creator>
<dc:date>2020-11-23</dc:date>
<dc:identifier>doi:10.1101/2020.11.23.394445</dc:identifier>
<dc:title><![CDATA[Regulation of Dopamine D1 Receptor Signaling at the Golgi Apparatus by a Low Affinity Dopamine Transporter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.24.395228v1?rss=1">
<title>
<![CDATA[
Sleep is required for odor exposure to consolidate memory and remodel olfactory synapses. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.24.395228v1?rss=1</link>
<description><![CDATA[
Animals with complex nervous systems demand sleep for memory consolidation and synaptic remodeling. Here we show that though the Caenorhabditis elegans nervous system has a limited number of neurons, sleep is necessary for both processes. In addition, it is unclear in any system if sleep collaborates with experience to alter synapses between specific neurons and whether this ultimately affects behavior. C. elegans neurons have defined connections and well-described contributions to behavior. We show that spaced odor-training and post-training sleep induce long-term memory. Memory consolidation, but not acquisition, requires a pair of interneurons, the AIYs, which play a role in odor-seeking behavior. In worms that consolidate memory, both sleep and odor conditioning are required to diminish inhibitory synaptic connections between the AWC chemosensory neurons and the AIYs. Thus, we demonstrate in a living organism that sleep is required for events immediately after training that drive memory consolidation and alter synaptic structures.
]]></description>
<dc:creator>Munoz-Lobato, F.</dc:creator>
<dc:creator>Benedetti, K. L.</dc:creator>
<dc:creator>Farah, F.</dc:creator>
<dc:creator>Chandra, R.</dc:creator>
<dc:creator>Bokka, A.</dc:creator>
<dc:creator>Brueggemann, C.</dc:creator>
<dc:creator>Saifuddin, F. M.</dc:creator>
<dc:creator>Nordquist, S. K.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Chang, E.</dc:creator>
<dc:creator>Varshney, A.</dc:creator>
<dc:creator>Jiminez, V.</dc:creator>
<dc:creator>Andersen, K. E.</dc:creator>
<dc:creator>Baradwaj, A.</dc:creator>
<dc:creator>Miller, J. M.</dc:creator>
<dc:creator>Dunn, R. L.</dc:creator>
<dc:creator>Tsujimoto, B.</dc:creator>
<dc:creator>Duong, A.</dc:creator>
<dc:creator>Tran, A.</dc:creator>
<dc:creator>Churgin, M.</dc:creator>
<dc:creator>Fang-Yen, C.</dc:creator>
<dc:creator>Bremer, M.</dc:creator>
<dc:creator>Kato, S.</dc:creator>
<dc:creator>VanHoven, M.</dc:creator>
<dc:creator>L'Etoile, N.</dc:creator>
<dc:date>2020-11-25</dc:date>
<dc:identifier>doi:10.1101/2020.11.24.395228</dc:identifier>
<dc:title><![CDATA[Sleep is required for odor exposure to consolidate memory and remodel olfactory synapses.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.27.400747v1?rss=1">
<title>
<![CDATA[
THE TRPV CHANNEL OSM-9 MEDIATES OLFACTORY LONG- TERM MEMORY CONSOLIDATION INDEPENDENT OF SLEEP 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.27.400747v1?rss=1</link>
<description><![CDATA[
Memory, defined as an alteration in behavior towards a stimulus that follows as a consequence of experience, arises when a sensory stimulus is encountered at the same time that the animal experiences a negative or positive internal state. How this coincident detection of external and internal stimuli stably alters responses to the external stimulus is still not fully understood, especially in the context of an intact animal. One barrier to understanding how an intact biological circuit changes is knowing what molecular processes are required to establish and maintain the memory. The optically accessible and compact nervous system of C. elegans provides a unique opportunity to examine these processes. C. elegans can remember an odor such as butanone when it is paired with a single negative experience and the transient receptor potential (TRP) OSM-9/TRPV5/TRPV6 channel is known to be required for this memory. The multiple gating mechanisms of TRPV channels give them the potential to be the coincidence detectors required to integrate internal state and external stimuli. Here, we report that this TRPV channel is also required for acquisition and possibly consolidation of sleep-dependent, long-term memory of butanone. We find that in the anterior ganglion, endogenous GFP-tagged OSM-9 is expressed in the paired AWA olfactory neurons, the ASH nociceptive neuron pair, the mechanosensory OLQ tetrad, and the paired ADF and ADL sensory neurons. In these cells, OSM-9 protein is concentrated in the sensory endings, dendrites, and cell bodies, but excluded from the neurites in the nerve ring. In the tail, OSM-9 is expressed in the nociceptive phasmid neurons PHA and PHB, possibly PQR as well as PVP. In the midbody, it is possibly expressed in the mechanosensitive PVD neuron. It is notably absent from the AWC pair that are required for butanone attraction. Chronic loss of OSM-9 in a subset of ciliated neurons that do not include AWA interferes with consolidation but not learning. Because OSM-9 is expressed and required in sensory neurons that are not needed for butanone chemosensory behavior, two interpretations are possible. The first, is that OSM-9 loss leads to gain of function or neomorphic behavior of these cells that are extrinsic to the primary sensory circuit and their new activity interferes with acquisition and consolidation of memory. The second is that loss of OSM-9 leads to a loss of function phenotype in which the wild type function of these cells is diminished and this function is required for memory consolidation.

Author summaryHow organisms learn from their environment and keep these memories for the long term ensures their survival. There is much known about the regions of the brain and the various proteins that are essential for memory, yet the exact molecular mechanisms and dynamics required are not known. We aimed to understand the genetics that underlie memory formation. We tested a gene that encodes a transient potential receptor channel vanilloid channel, which is similar to the channels we have that sense spicy foods and other harmful cues. Our studies have shown that this gene is required for the animal to be able to acquire and perhaps consolidate olfactory memory. This protein is not expressed in the sensory neurons that respond to the odor that is memorized or in other downstream interneurons in the odor-sensation circuit, but it is expressed in a distinct set of sensory neurons. This indicates that long-term memory involves wild type behavior of a wider array of sensory neurons than is required for the primary sensation. These channels are also implicated in neurological disorders where memory is affected, including Alzheimers disease. Understanding how memory formation is affected by cells outside the memory circuit might provide testable hypothesis about what goes awry in Alzheimers disease.
]]></description>
<dc:creator>Benedetti, K. L.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>L'Etoile, N.</dc:creator>
<dc:date>2020-11-29</dc:date>
<dc:identifier>doi:10.1101/2020.11.27.400747</dc:identifier>
<dc:title><![CDATA[THE TRPV CHANNEL OSM-9 MEDIATES OLFACTORY LONG- TERM MEMORY CONSOLIDATION INDEPENDENT OF SLEEP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.30.401885v1?rss=1">
<title>
<![CDATA[
Lyophilized yeast powder for adjuvant free thermostable vaccine delivery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.30.401885v1?rss=1</link>
<description><![CDATA[
Thermolabile nature of commercially available vaccines necessitates their storage, transportation and dissemination under refrigerated condition. Maintenance of continuous cold chain at every step increases the final cost of vaccines. Any breach in the cold chain, even for a short duration results in the need to discard the vaccine. As a result, there is a pressing need for the development of thermostable vaccines. In this proof of concept study, we showed that E. coli curli-GFP fusion protein remains stable in freeze-dried yeast powder for more than a 13 and 6 months when stored at 30 {degrees}C and 37 {degrees}C respectively. Stability of the heterologous protein remains unaffected during the process of heat-inactivation and lyophilization. The mass of lyophilized yeast powder remains almost unchanged during the entire period of storage. Expressed protein remains intact even after two cycles of freeze and thaws. The protease deficient strain appears ideal for the development of whole recombinant yeast-based vaccines. The cellular abundance of expressed antigen in dry powder after a year was comparable to freshly lyophilized cells. SEM microscopy showed the intact nature of cells in powdered form even after a year of storage at 30 {degrees}C. Observation made in this study showed that freeze-dry yeast powder can play a vital role in the development of thermostable vaccines.
]]></description>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Kharbikar, B. N.</dc:creator>
<dc:creator>Desai, T. A.</dc:creator>
<dc:date>2020-12-01</dc:date>
<dc:identifier>doi:10.1101/2020.11.30.401885</dc:identifier>
<dc:title><![CDATA[Lyophilized yeast powder for adjuvant free thermostable vaccine delivery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.03.410837v1?rss=1">
<title>
<![CDATA[
Zika Virus Infection Prevents Host mRNA Nuclear Export by Disrupting UPF1 Function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.03.410837v1?rss=1</link>
<description><![CDATA[
Zika virus (ZIKV) is a mosquito-borne RNA virus that can infect fetuses in utero causing characteristic neurodevelopmental disorders including microcephaly. We previously showed that ZIKV infection downregulates expression of up-frameshift protein 1 (UPF1), a helicase/ATPase and central regulator of the nonsense-mediated mRNA decay pathway. Here, we identify a novel function of nuclear UPF1 in mRNA export. Using crosslinking immunoprecipitation of UPF1 followed by sequencing of associated transcripts as well as fluorescence in situ hybridization experiments, we find widespread mRNA accumulation in the nucleus of human neural progenitor cells (NPCs) upon ZIKV infection or UPF1 knockdown. Knockdown of FREM2, a top UPF1 target transcript encoding an extra-cellular matrix protein critical in fetal development, decreased expression of pluripotency markers and increased expressed neuronal differentiation in NPCs, consistent with the model that trapping FREM2 mRNA in the nucleus perturbs proper NPC function. Collectively, our data uncover a new posttranscriptional mechanism by which ZIKV "shuts off" host mRNA export via UPF1. As we find UPF1 linked to many neurodevelopment pathways, we propose that the lack of host mRNA export contributes to the neurodevelopmental defects associated with ZIKV infection.
]]></description>
<dc:creator>Leon, K. E.</dc:creator>
<dc:creator>Flynn, R. E.</dc:creator>
<dc:creator>Khalid, M. M.</dc:creator>
<dc:creator>Fontaine, K. A.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Simoneau, C. R.</dc:creator>
<dc:creator>Tomar, S.</dc:creator>
<dc:creator>Jimenez-Morales, D.</dc:creator>
<dc:creator>Dunlap, M.</dc:creator>
<dc:creator>Kaye, J. A.</dc:creator>
<dc:creator>Shah, P. S.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Bertozzi, C.</dc:creator>
<dc:creator>Carette, J. E.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:date>2020-12-04</dc:date>
<dc:identifier>doi:10.1101/2020.12.03.410837</dc:identifier>
<dc:title><![CDATA[Zika Virus Infection Prevents Host mRNA Nuclear Export by Disrupting UPF1 Function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.06.413849v1?rss=1">
<title>
<![CDATA[
Starvation induces shrinkage of the bacterial cytoplasm 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.06.413849v1?rss=1</link>
<description><![CDATA[
Environmental fluctuations are a common challenge for single-celled organisms; enteric bacteria such as Escherichia coli experience dramatic changes in nutrient availability, pH, and temperature during their journey into and out of the host. While the effects of altered nutrient availability on gene expression and protein synthesis are well known, their impacts on cytoplasmic dynamics and cell morphology have been largely overlooked. Here, we discover that depletion of utilizable nutrients results in shrinkage of E. colis inner membrane from the cell wall. Shrinkage was accompanied by a [~]17% reduction in cytoplasmic volume and a concurrent increase in periplasmic volume. Inner membrane retraction occurred almost exclusively at the new cell pole. This phenomenon was distinct from turgor-mediated plasmolysis and independent of new transcription, translation, or canonical starvation-sensing pathways. Cytoplasmic dry-mass density increased during shrinkage, suggesting that it is driven primarily by loss of water. Shrinkage was reversible: upon a shift to nutrient-rich medium, expansion started almost immediately at a rate dependent on carbon-source quality. Robust recovery from starvation required the Tol-Pal system, highlighting the importance of envelope coupling during recovery. Klebsiella pneumoniae also exhibited shrinkage when shifted to carbon-free conditions, suggesting a conserved phenomenon. These findings demonstrate that even when Gram-negative bacterial growth is arrested, cell morphology and physiology are still dynamic.

Significance statementBacterial cells constantly face nutrient fluctuations in their natural environments. While previous studies have identified gene expression changes upon nutrient depletion, it is much less well known how cellular morphology and cytoplasmic properties respond to shifts in nutrient availability. Here, we discovered that switching fast-growing Escherichia coli cells to nutrient-free conditions results in substantial shrinkage of the inner membrane away from the cell wall, especially at the new pole. Shrinkage was primarily driven by loss of cytoplasmic water contents. Shrinkage was also exhibited by cells naturally entering stationary phase, highlighting its biological relevance across starvation conditions. The membrane-spanning Tol-Pal system was critical for robust entry into and recovery from shrinkage, indicating the importance of cell-envelope homeostasis in surviving nutrient starvation.
]]></description>
<dc:creator>Shi, H.</dc:creator>
<dc:creator>Westfall, C. S.</dc:creator>
<dc:creator>Kao, J.</dc:creator>
<dc:creator>Odermatt, P. D.</dc:creator>
<dc:creator>Cesar, S.</dc:creator>
<dc:creator>Sievert, M.</dc:creator>
<dc:creator>Moore, J.</dc:creator>
<dc:creator>Gonzalez, C. G.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Elias, J. E.</dc:creator>
<dc:creator>Chang, F.</dc:creator>
<dc:creator>Huang, K. C.</dc:creator>
<dc:creator>Levin, P. A.</dc:creator>
<dc:date>2020-12-07</dc:date>
<dc:identifier>doi:10.1101/2020.12.06.413849</dc:identifier>
<dc:title><![CDATA[Starvation induces shrinkage of the bacterial cytoplasm]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.13.422538v1?rss=1">
<title>
<![CDATA[
Deep Linear Modeling of Hierarchical Functional Connectivity in the Human Brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.13.422538v1?rss=1</link>
<description><![CDATA[
The human brain exhibits hierarchical modular organization, which is not depicted by conventional fMRI functional connectivity reconstruction methods such as independent component analysis (ICA). To map hierarchical brain connectivity networks (BCNs), we propose a novel class of deep (multilayer) linear models that are constructed such that each successive layer decomposes the features of the preceding layer. Three of these are multilayer variants of Sparse Dictionary Learning (SDL), Non-Negative Matrix Factorization (NMF) and Fast ICA (FICA). We present a fourth deep linear model, Deep Matrix Fitting (MF), which incorporates both rank reduction for data-driven hyperparameter determination as well as a distributed optimization function. We also introduce a novel framework for theoretical comparison of these deep linear models based on their combination of mathematical operators, the predictions of which are tested using simulated resting state fMRI data with known ground truth BCNs. Consistent with the theoretical predictions, Deep MF and Deep SDL performed best for connectivity estimation of 1st layer networks, whereas Deep FICA and Deep NMF were modestly better for spatial mapping. Deep MF provided the best overall performance, including computational speed. These deep linear models can efficiently map hierarchical BCNs without requiring the manual hyperparameter tuning, extensive fMRI training data or high-performance computing infrastructure needed by deep nonlinear models, such as convolutional neural networks (CNNs) or deep belief networks (DBNs), and their results are also more explainable from their mathematical structure. These benefits gain in importance as continual improvements in the spatial and temporal resolution of fMRI reveal more of the hierarchy of spatiotemporal brain architecture. These new models of hierarchical BCNs may also advance the development of fMRI diagnostic and prognostic biomarkers, given the recent recognition of disparities between low-level vs high-level network connectivity across a wide range of neurological and psychiatric disorders.
]]></description>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Palacios, E. M.</dc:creator>
<dc:creator>Mukherjee, P.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.13.422538</dc:identifier>
<dc:title><![CDATA[Deep Linear Modeling of Hierarchical Functional Connectivity in the Human Brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.13.422611v1?rss=1">
<title>
<![CDATA[
How inhibitory neurons increase information transmission under threshold modulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.13.422611v1?rss=1</link>
<description><![CDATA[
Modulation of neuronal thresholds is ubiquitous in the brain. Phenomena such as figure-ground segmentation, motion detection, stimulus anticipation and shifts in attention all involve changes in a neurons threshold based on signals from larger scales than its primary inputs. However, this modulation reduces the accuracy with which neurons can represent their primary inputs, creating a mystery as to why threshold modulation is so widespread in the brain. We find that modulation is less detrimental than other forms of neuronal variability and that its negative effects can be nearly completely eliminated if modulation is applied selectively to sparsely responding neurons in a circuit by inhibitory neurons. We verify these predictions in the retina where we find that inhibitory amacrine cells selectively deliver modulation signals to sparsely responding ganglion cell types. Our findings elucidate the central role that inhibitory neurons play in maximizing information transmission under modulation.
]]></description>
<dc:creator>Hsu, W.-M. M.</dc:creator>
<dc:creator>Kastner, D. B.</dc:creator>
<dc:creator>Baccus, S. A.</dc:creator>
<dc:creator>Sharpee, T. O.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.13.422611</dc:identifier>
<dc:title><![CDATA[How inhibitory neurons increase information transmission under threshold modulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.14.422686v1?rss=1">
<title>
<![CDATA[
Juvenile exposure to acute traumatic stress leads to long-lasting alterations in grey matter myelination in adult female but not male rats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.14.422686v1?rss=1</link>
<description><![CDATA[
Stress early in life can have a major impact on brain development, and there is increasing evidence that childhood stress confers vulnerability for later developing psychiatric disorders. In particular, during peri-adolescence, brain regions crucial for emotional regulation, such as the prefrontal cortex (PFC), amygdala (AMY) and hippocampus (HPC), are still developing and are highly sensitive to stress. Changes in myelin levels have been implicated in mental illnesses and stress effects on myelin and oligodendrocytes (OLs) are beginning to be explored as a novel and underappreciated mechanism underlying psychopathologies. Yet there is little research on the effects of acute stress on myelin during peri-adolescence, and even less work exploring sex-differences. Here, we used a rodent model to test the hypothesis that exposure to acute traumatic stress as a juvenile would induce changes in OLs and myelin content across limbic brain regions. Male and female juvenile rats underwent three hours of restraint stress with exposure to a predator odor on postnatal day (p) 28. Acute stress induced a physiological response, increasing corticosterone release and reducing weight gain in stress-exposed animals. Brain sections containing the PFC, AMY and HPC were taken either in adolescence (p40), or in adulthood (p95) and stained for markers of OLs and myelin. We found that acute stress induced sex-specific changes in grey matter (GM) myelination and OLs in both the short- and long-term. Exposure to a single stressor as a juvenile increased GM myelin content in the AMY and HPC in p40 males, compared to the respective control group. At p40, corticosterone release during stress exposure was also positively correlated with GM myelin content in the AMY of male rats. Single exposure to juvenile stress also led to long-term effects exclusively in female rats. Compared to controls, stress-exposed females showed reduced GM myelin content in all three brain regions. Acute stress exposure decreased PFC and HPC OL density in p40 females, perhaps contributing towards this observed long-term decrease in myelin content. Overall, our findings suggest that the juvenile brain is vulnerable to exposure to a brief severe stressor. Exposure to a single short traumatic event during peri-adolescence produces long-lasting changes in GM myelin content in the adult brain of female, but not male, rats. These findings highlight myelin plasticity as a potential contributor to sex-specific sensitivity to perturbation during a critical window of development.
]]></description>
<dc:creator>Breton, J. M.</dc:creator>
<dc:creator>Barraza, M.</dc:creator>
<dc:creator>Hu, K. Y.</dc:creator>
<dc:creator>Frias, S. J.</dc:creator>
<dc:creator>Long, K. L. P.</dc:creator>
<dc:creator>Kaufer, D.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.14.422686</dc:identifier>
<dc:title><![CDATA[Juvenile exposure to acute traumatic stress leads to long-lasting alterations in grey matter myelination in adult female but not male rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.15.412809v1?rss=1">
<title>
<![CDATA[
Cyclic gallium-68 labeled peptides for specific detection of human angiotensin-converting enzyme 2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.15.412809v1?rss=1</link>
<description><![CDATA[
In this study, we developed ACE2-specific, peptide-derived 68Ga-labeled radiotracers, motivated by the hypotheses that (1) ACE2 is an important determinant of SARS-CoV-2 susceptibility, and (2) that modulation of ACE2 in COVID-19 drives severe organ injury.

MethodsA series of NOTA-conjugated peptides derived from the known ACE2 inhibitor DX600 were synthesized, with variable linker identity. Since DX600 bears two cystine residues, both linear and cyclic peptides were studied. An ACE2 inhibition assay was used to identify lead compounds, which were labeled with 68Ga to generate peptide radiotracers ([68Ga]NOTA-PEP). The aminocaproate-derived radiotracer [68Ga]NOTA-PEP4 was subsequently studied in a humanized ACE2 (hACE2) transgenic model.

ResultsCyclic DX-600 derived peptides had markedly lower IC50s than their linear counterparts. The three cyclic peptides with triglycine, aminocaproate, and polyethylene glycol linkers had calculated IC50s similar to, or lower than the parent DX600 molecule. Peptides were readily labeled with 68Ga, and the biodistribution of [68Ga]NOTA-PEP4 was determined in a hACE2 transgenic murine cohort. Pharmacologic concentrations of co-administered NOTA-PEP ("blocking") showed significant reduction of [68Ga]NOTA-PEP4 signals in the in the heart, liver, lungs, and small intestine. Ex vivo hACE2 activity in these organs was confirmed as a correlate to in vivo results.

ConclusionsNOTA-conjugated, cyclic peptides derived from the known ACE2 inhibitor DX600 retain their activity when N-conjugated for 68Ga chelation. In vivo studies in a transgenic hACE2 murine model using the lead tracer [68Ga]NOTA-PEP4 showed specific binding in the heart, liver, lungs and intestine - organs known to be affected in SARS-CoV-2 infection. These results suggest that [68Ga]NOTA-PEP4 could be used to detect organ-specific suppression of ACE2 in SARS-CoV-2 infected murine models and COVID-19 patients.

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]]></description>
<dc:creator>Parker, M.</dc:creator>
<dc:creator>Blecha, J.</dc:creator>
<dc:creator>Rosenberg, O.</dc:creator>
<dc:creator>Ohliger, M.</dc:creator>
<dc:creator>Flavell, R.</dc:creator>
<dc:creator>Wilson, D. M.</dc:creator>
<dc:date>2020-12-16</dc:date>
<dc:identifier>doi:10.1101/2020.12.15.412809</dc:identifier>
<dc:title><![CDATA[Cyclic gallium-68 labeled peptides for specific detection of human angiotensin-converting enzyme 2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.15.422949v1?rss=1">
<title>
<![CDATA[
Microbiome and Metabolome driven differentiation of TGF-β producing Tregs leads to Senescence and HIV latency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.15.422949v1?rss=1</link>
<description><![CDATA[
Current therapeutic interventions to eradicate latent HIV ("reservoir") and restore immune function in ART-treated HIV infection have yet to show efficacy. To explore mechanisms of HIV persistence, we apply an integrated systems biology approach and identify a distinct group of individuals with poor CD4 T-cell reconstitution (Immunologic non-responders, "INRs") and high frequencies of cells with inducible HIV. Contrary to the prevailing notion that immune activation drives HIV persistence and immune dysfunction, peripheral blood leukocytes from these subjects have enhanced expression of a network of genes regulated by cellular senescence driving transcription factors (TFs) FOXO3, SMAD2 and IRF3. In these subjects, increased frequencies of regulatory T cells and expression of the TGF-{beta} signaling cascade are complimented by the downregulation of cell cycle, metabolic and pro-inflammatory pathways. Lactobacillaceae family and metabolites (members of the butyrate family - i.e. -ketobutyrate) were correlated with Treg frequencies in "Senescent-INRs" ex vivo, triggered the differentiation of TGF-{beta} producing Tregs and promoted HIV latency establishment in vitro. These cascades, downstream of PD-1/TGF-{beta}, prevent memory T cell differentiation and are associated with an increase in frequencies of cells with inducible HIV ex vivo. Our findings identify cellular senescence responses that can be targeted by PD-1 or TGF-{beta} specific interventions that have shown safety and efficacy in cancer, and may prove to be crucial for HIV eradication.
]]></description>
<dc:creator>Sekaly, R.-P.</dc:creator>
<dc:creator>Ghneim, K.</dc:creator>
<dc:creator>Sharma, A. A.</dc:creator>
<dc:creator>Ribeiro, S. P.</dc:creator>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Ahlers, J.</dc:creator>
<dc:creator>Kulpa, D.</dc:creator>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Brehm, J.</dc:creator>
<dc:creator>Talla, A.</dc:creator>
<dc:creator>Arumugam, S.</dc:creator>
<dc:creator>Darko, S.</dc:creator>
<dc:creator>Rodriguez, B.</dc:creator>
<dc:creator>Shive, C.</dc:creator>
<dc:creator>Cristescu, R.</dc:creator>
<dc:creator>Loboda, A.</dc:creator>
<dc:creator>Balderas, R.</dc:creator>
<dc:creator>Wang, I.-m.</dc:creator>
<dc:creator>Hunt, P.</dc:creator>
<dc:creator>Lamarre, D.</dc:creator>
<dc:creator>Douek, D.</dc:creator>
<dc:creator>Hazuda, D.</dc:creator>
<dc:creator>Lederman, M.</dc:creator>
<dc:creator>Deeks, S.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.15.422949</dc:identifier>
<dc:title><![CDATA[Microbiome and Metabolome driven differentiation of TGF-β producing Tregs leads to Senescence and HIV latency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.17.423129v1?rss=1">
<title>
<![CDATA[
Broad transcriptomic dysregulation across the cerebral cortex in ASD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.17.423129v1?rss=1</link>
<description><![CDATA[
Classically, psychiatric disorders have been considered to lack defining pathology, but recent work has demonstrated consistent disruption at the molecular level, characterized by transcriptomic and epigenetic alterations.1-3 In ASD, upregulation of microglial, astrocyte, and immune signaling genes, downregulation of specific synaptic genes, and attenuation of regional gene expression differences are observed.1,2,4-6 However, whether these changes are limited to the cortical association areas profiled is unknown. Here, we perform RNA-sequencing (RNA-seq) on 725 brain samples spanning 11 distinct cortical areas in 112 ASD cases and neurotypical controls. We identify substantially more genes and isoforms that differentiate ASD from controls than previously observed. These alterations are pervasive and cortex-wide, but vary in magnitude across regions, roughly showing an anterior to posterior gradient, with the strongest signal in visual cortex, followed by parietal cortex and the temporal lobe. We find a notable enrichment of ASD genetic risk variants among cortex-wide downregulated synaptic plasticity genes and upregulated protein folding gene isoforms. Finally, using snRNA-seq, we determine that regional variation in the magnitude of transcriptomic dysregulation reflects changes in cellular proportion and cell-type-specific gene expression, particularly impacting L3/4 excitatory neurons. These results highlight widespread, genetically-driven neuronal dysfunction as a major component of ASD pathology in the cerebral cortex, extending beyond association cortices to involve primary sensory regions.
]]></description>
<dc:creator>Haney, J. R.</dc:creator>
<dc:creator>Wamsley, B.</dc:creator>
<dc:creator>Chen, G. T.</dc:creator>
<dc:creator>Parhami, S.</dc:creator>
<dc:creator>Emani, P. S.</dc:creator>
<dc:creator>Chang, N.</dc:creator>
<dc:creator>Hoftman, G. D.</dc:creator>
<dc:creator>de Alba, D.</dc:creator>
<dc:creator>Kale, G.</dc:creator>
<dc:creator>Ramaswami, G.</dc:creator>
<dc:creator>Hartl, C. L.</dc:creator>
<dc:creator>Jin, T.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Ou, J.</dc:creator>
<dc:creator>Wu, Y. E.</dc:creator>
<dc:creator>Parikshak, N. N.</dc:creator>
<dc:creator>Swarup, V.</dc:creator>
<dc:creator>Belgard, T. G.</dc:creator>
<dc:creator>Gerstein, M.</dc:creator>
<dc:creator>Pasaniuc, B.</dc:creator>
<dc:creator>Gandal, M. J.</dc:creator>
<dc:creator>Geschwind, D. H.</dc:creator>
<dc:date>2020-12-18</dc:date>
<dc:identifier>doi:10.1101/2020.12.17.423129</dc:identifier>
<dc:title><![CDATA[Broad transcriptomic dysregulation across the cerebral cortex in ASD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.18.423526v1?rss=1">
<title>
<![CDATA[
Interneuron origins in the embryonic porcine medial ganglionic eminence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.18.423526v1?rss=1</link>
<description><![CDATA[
Interneurons contribute to the complexity of neural circuits and maintenance of normal brain function. Rodent interneurons originate in embryonic ganglionic eminences, but developmental origins in other species are less understood. Here, we show that transcription factor expression patterns in porcine embryonic subpallium are similar to rodents, delineating a distinct medial ganglionic eminence (MGE) progenitor domain. On the basis of Nkx2.1, Lhx6 and Dlx2 expression, in vitro differentiation into neurons expressing GABA and robust migratory capacity in explant assays, we propose that cortical and hippocampal interneurons originate from a porcine MGE region. Following xenotransplantation into adult male and female rat hippocampus, we further demonstrate that porcine MGE progenitors, like those from rodents, migrate and differentiate into morphologically distinct interneurons expressing GABA. Our findings reveal that basic rules for interneuron development are conserved across species, and that porcine embryonic MGE progenitors could serve as a valuable source for interneuron-based xenotransplantation therapies.

Significance StatementHere we demonstrate that porcine MGE, like rodents, exhibit a distinct transcriptional and pallial interneuron-specific antibody profile, in vitro migratory capacity and are amenable to xenotransplantation. This is the first comprehensive examination of embryonic pallial interneuron origins in the pig, and because a rich neurodevelopmental literature on embryonic mouse MGE exists (with some additional characterizations in other species like monkey and human) our work allows direct neurodevelopmental comparisons with this literature.
]]></description>
<dc:creator>Casalia, M. L.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Ramsay, H.</dc:creator>
<dc:creator>ROSS, P. J.</dc:creator>
<dc:creator>Paredes, M. F.</dc:creator>
<dc:creator>Baraban, S. C.</dc:creator>
<dc:date>2020-12-21</dc:date>
<dc:identifier>doi:10.1101/2020.12.18.423526</dc:identifier>
<dc:title><![CDATA[Interneuron origins in the embryonic porcine medial ganglionic eminence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.20.423692v1?rss=1">
<title>
<![CDATA[
CTCF is a Barrier for Totipotent-like Reprogramming 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.20.423692v1?rss=1</link>
<description><![CDATA[
Totipotent cells have the ability of generating embryonic and extra-embryonic tissues1,2. Interestingly, a rare population of cells with totipotent-like potential was identified within ESC cultures3. These cells, known as 2 cell (2C)-like cells, arise from ESC and display similar features to those found in the totipotent 2 cell embryo2-4. However, the molecular determinants of 2C-like conversion have not been completely elucidated. Here, we show that CTCF is a barrier for 2C-like reprogramming. Indeed, forced conversion to a 2C-like state by DUX expression was associated with DNA damage at a subset of CTCF binding sites. Endogenous or DUX-induced 2C-like ESC showed decreased CTCF enrichment at known binding sites, suggesting that acquisition of a totipotent-like state is associated with a highly dynamic chromatin architecture. Accordingly, depletion of CTCF in ESC efficiently promoted spontaneous and asynchronous conversion to a totipotent-like state. This phenotypic reprogramming was reversible upon restoration of CTCF levels. Furthermore, we showed that transcriptional activation of the ZSCAN4 cluster was necessary for successful 2C-like reprogramming. In summary, we revealed the intimate relation between CTCF and totipotent-like reprogramming.
]]></description>
<dc:creator>Olbrich, T.</dc:creator>
<dc:creator>Vega-Sendino, M.</dc:creator>
<dc:creator>Tillo, D.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Zolnerowich, N.</dc:creator>
<dc:creator>Tran, A. D.</dc:creator>
<dc:creator>Domingo, C. N.</dc:creator>
<dc:creator>Franco, M.</dc:creator>
<dc:creator>Markiewicz Potoczny, M.</dc:creator>
<dc:creator>Pegoraro, G.</dc:creator>
<dc:creator>Fitzgerald, P.</dc:creator>
<dc:creator>Kruhlak, M.</dc:creator>
<dc:creator>Lazzerini, E.</dc:creator>
<dc:creator>Nora, E.</dc:creator>
<dc:creator>Nussenzweig, A.</dc:creator>
<dc:creator>Ruiz Macias, S.</dc:creator>
<dc:date>2020-12-22</dc:date>
<dc:identifier>doi:10.1101/2020.12.20.423692</dc:identifier>
<dc:title><![CDATA[CTCF is a Barrier for Totipotent-like Reprogramming]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.21.423397v1?rss=1">
<title>
<![CDATA[
Multicohort Analysis of Publicly-available Monocyte Expression Data Identifies Gene Signatures to Accurately Monitor Subset-specific Changes in Human Diseases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.21.423397v1?rss=1</link>
<description><![CDATA[
Monocytes and monocyte-derived cells play important roles in the regulation of inflammation, both as precursors as well as effector cells. Monocytes are heterogeneous and characterized by three distinct subsets in humans. Classical and non-classical monocytes represent the most abundant subsets, each carrying out distinct biological functions. Consequently, altered frequencies of different subsets have been associated with inflammatory conditions, such as infections and autoimmune disorders including lupus, rheumatoid arthritis, inflammatory bowel disease, and, more recently, COVID-19. Dissecting the contribution of different monocyte subsets to disease is currently limited by samples and cohorts, often resulting in underpowered studies and, consequently, poor reproducibility. Public transcriptomes provide an alternative source of data characterized by high statistical power and real world heterogeneity. However, most transcriptome datasets profile bulk blood or tissue samples, requiring the use of in silico approaches to quantify changes in the levels of specific cell types.

Here, we integrated 853 publicly available microarray expression profiles of sorted human monocyte subsets from 45 independent studies to identify robust and parsimonious gene expression signatures, consisting of 10 genes specific to each subset. These signatures, although derived using only datasets profiling healthy individuals, maintain their accuracy independent of the disease state in an independent cohort profiled by RNA-sequencing (AUC = 1.0). Furthermore, we demonstrate that our signatures are specific to monocyte subsets compared to other immune cells such as B, T, dendritic cells (DCs) and natural killer (NK) cells (AUC = 0.87~0.88, p<2.2e-16). This increased specificity results in estimated monocyte subset levels that are strongly correlated with cytometry-based quantification of cellular subsets (r = 0.69, p = 6.7e-4). Consequently, we show that these monocyte subset-specific signatures can be used to quantify changes in monocyte subsets levels in expression profiles from patients in clinical trials. Finally, we show that proteins encoded by our signature genes can be used in cytometry-based assays to specifically sort monocyte subsets. Our results demonstrate the robustness, versatility, and utility of our computational approach and provide a framework for the discovery of new cellular markers.
]]></description>
<dc:creator>Vallania, F.</dc:creator>
<dc:creator>Zisman, L.</dc:creator>
<dc:creator>Macaubas, C.</dc:creator>
<dc:creator>Hung, S.-C.</dc:creator>
<dc:creator>Rajasekaran, N.</dc:creator>
<dc:creator>Mason, S.</dc:creator>
<dc:creator>Graf, J.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Mellins, E.</dc:creator>
<dc:creator>Khatri, P.</dc:creator>
<dc:date>2020-12-22</dc:date>
<dc:identifier>doi:10.1101/2020.12.21.423397</dc:identifier>
<dc:title><![CDATA[Multicohort Analysis of Publicly-available Monocyte Expression Data Identifies Gene Signatures to Accurately Monitor Subset-specific Changes in Human Diseases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.22.423985v1?rss=1">
<title>
<![CDATA[
CRISPR-based functional genomics in human dendritic cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.22.423985v1?rss=1</link>
<description><![CDATA[
Dendritic cells (DCs) regulate processes ranging from antitumor and antiviral immunity to host-microbe communication at mucosal surfaces. It remains difficult, however, to genetically manipulate human DCs, limiting our ability to probe how DCs elicit specific immune responses. Here, we develop a CRISPR/Cas9 genome editing method for human monocyte-derived DCs (moDCs) that mediates knockouts with a median efficiency of >93% across >300 genes. Using this method, we perform genetic screens in moDCs, identifying mechanisms by which DCs tune responses to lipopolysaccharides from the human microbiome. In addition, we reveal donor-specific responses to lipopolysaccharides, underscoring the importance of assessing immune phenotypes in donor-derived cells, and identify genes that control this specificity, highlighting the potential of our method to pinpoint determinants of inter-individual variation in immune responses. Our work sets the stage for a systematic dissection of the immune signaling at the host-microbiome interface and for targeted engineering of DCs for neoantigen vaccination.
]]></description>
<dc:creator>Jost, M.</dc:creator>
<dc:creator>Jacobson, A. N.</dc:creator>
<dc:creator>Hussmann, J. A.</dc:creator>
<dc:creator>Cirolia, G.</dc:creator>
<dc:creator>Fischbach, M.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:date>2020-12-22</dc:date>
<dc:identifier>doi:10.1101/2020.12.22.423985</dc:identifier>
<dc:title><![CDATA[CRISPR-based functional genomics in human dendritic cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.22.424036v1?rss=1">
<title>
<![CDATA[
Elucidation of the molecular interactions that enable stable assembly and structural diversity in multicomponent immune receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.22.424036v1?rss=1</link>
<description><![CDATA[
Multi-component immune receptors are essential complexes in which distinct ligand-recognition and signaling subunits are held together by interactions between acidic and basic residues of their transmembrane helices. A 2-to-1 acidic to basic motif in the transmembrane domains of the subunits is necessary and sufficient to assemble these receptor complexes. Here, we study a prototype for these receptors, a DAP12-NKG2C 2:1 heterotrimeric complex, in which the two DAP12 subunits each contribute a single transmembrane Asp residue, and the NKG2C subunit contributes a Lys to form the complex. DAP12 can also associate with 20 other subunits using a similar motif. Here we use molecular dynamics simulations to understand the basis for the high affinity and diversity of interactions in this group of receptors. Simulations of the transmembrane helices with differing protonation states of the Asp-Asp-Lys triad identified a structurally stable interaction in which a singly protonated Asp-Asp pair forms a hydrogen-bonded carboxyl-carboxylate clamp that clasps onto a charged Lys sidechain. This polar motif was also supported by density functional theory and a Protein Data Bank-wide search. In contrast, the helices are dynamic at sites distal to the stable carboxyl-carboxylate clamp motif. Such a locally stable but globally dynamic structure is well-suited to accommodate the sequence and structural variations in the transmembrane helices of multi-component receptors, which mix and match subunits to create combinatorial functional diversity from a limited number of subunits. It also supports a signaling mechanism based on multi-subunit clustering rather than propagation of rigid conformational changes through the membrane.

SIGNIFICANCEReceptors that separate ligand recognition and intracellular signaling into separate protein subunits are ubiquitous in immunity. These subunits mix and match to create combinatorial functional diversity. The transmembrane domains of these receptors assemble through the interaction between two acidic and one basic residue on different helices. Using computational methods to study the DAP12-NKG2C receptor complex we identified a polar motif in which a singly protonated Asp-Asp pair forms a carboxyl-carboxylate clamp that clasps a charged Lys sidechain in the membrane. This local interaction allows dynamic variations in other regions of the helices that tolerate sequence diversity of the interacting subunits in this class of receptors, which signal through multi-subunit clustering rather than propagation of rigid conformational changes through the membrane.
]]></description>
<dc:creator>Fong, L.-K.</dc:creator>
<dc:creator>Chalkely, M. J.</dc:creator>
<dc:creator>Tan, S. K.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>DeGrado, W.</dc:creator>
<dc:date>2020-12-22</dc:date>
<dc:identifier>doi:10.1101/2020.12.22.424036</dc:identifier>
<dc:title><![CDATA[Elucidation of the molecular interactions that enable stable assembly and structural diversity in multicomponent immune receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.23.423300v1?rss=1">
<title>
<![CDATA[
Selective Targeting of Ras Mutant Cancers via a New Small Molecule 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.23.423300v1?rss=1</link>
<description><![CDATA[
Mutations in the Ras family of oncogenes are implicated in 33% of human cancers, making Ras an intensely pursued target in drug discovery. As an alternative to direct pharmacological inhibition of Ras, we looked for sensitivities in RAS mutant cells. Using a small molecule screen in cell lines with mutations in Ras and its effector Raf, we discovered 249C as a Ras-mutant selective cytotoxic agent against a spectrum of RAS-mutant cancers. By combining CRISPR chemical-genetic screening, comparative profiling and chemoproteomics, we identified that 249C binds to a unique subunit on vacuolar (V)-ATPase with nanomolar affinity, inhibiting its biochemical activity and, unexpectedly, altering V-ATPase translocation in Ras-induced macropinocytosis. Via binding to V-ATPase, 249C prevents lysosomal acidification and inhibits autophagy and macropinocytosis pathways that several Ras-driven cancers rely on for survival. In characterizing 249Cs mechanism, we show that potency varies with the identity of the RAS driver mutation highlighting a mutant-specific dependence on autophagy and macropinocytosis. Indeed, 249C potently inhibits tumor growth without adverse side effects in a mouse xenograft model of KRAS-driven non-small cell lung cancer. These data establish proof-of-concept for targeting V-ATPase as a way to indirectly target specific Ras mutants, and provide a fundamental link between V-ATPase localization and specific Ras mutant tumor-related activity.
]]></description>
<dc:creator>Tolani, B.</dc:creator>
<dc:creator>Celli, A.</dc:creator>
<dc:creator>Yao, Y.</dc:creator>
<dc:creator>Tan, Y. Z.</dc:creator>
<dc:creator>Fetter, R.</dc:creator>
<dc:creator>Liem, C. R.</dc:creator>
<dc:creator>de Smith, A. J.</dc:creator>
<dc:creator>Vasanthakumar, T.</dc:creator>
<dc:creator>Bisignano, P.</dc:creator>
<dc:creator>Seiple, I. B.</dc:creator>
<dc:creator>Rubinstein, J. L.</dc:creator>
<dc:creator>Jost, M.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:date>2020-12-23</dc:date>
<dc:identifier>doi:10.1101/2020.12.23.423300</dc:identifier>
<dc:title><![CDATA[Selective Targeting of Ras Mutant Cancers via a New Small Molecule]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.25.424409v1?rss=1">
<title>
<![CDATA[
Negative feedback by NUR77/Nr4a1 restrains B cell clonal dominance during early T-dependent immune responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.25.424409v1?rss=1</link>
<description><![CDATA[
B cell clones compete for entry into and dominance within germinal centers (GC), where the highest affinity BCRs are selected. However, diverse and low affinity B cells can enter and reside in GCs for extended periods. To reconcile these observations, we hypothesized that a negative feedback loop may operate within B cells to preferentially restrain high affinity clones from monopolizing the early GC niche. Here we report a role for the nuclear receptor NUR77/Nr4a1 in this process. We previously showed that NUR77 expression scales with antigen stimulation and restrains B cell expansion when T cell help is limiting. Here we show that, although NUR77 is dispensable for regulating GC size when GC are elicited in a largely clonal manner, it serves to curb immunodominance under conditions where diverse clonal populations must compete for a constrained niche. Moreover, this is independent of B cell precursor frequency and reflects, at least in part, a B cell-intrinsic role for NUR77. We propose that this is important to preserve early B cell clonal diversity in order to limit holes in the post-immune repertoire and to optimize GC selection.
]]></description>
<dc:creator>Brooks, J. F.</dc:creator>
<dc:creator>Tan, C.</dc:creator>
<dc:creator>Mueller, J. L.</dc:creator>
<dc:creator>Hibiya, K.</dc:creator>
<dc:creator>Hiwa, R.</dc:creator>
<dc:creator>Zikherman, J.</dc:creator>
<dc:date>2020-12-26</dc:date>
<dc:identifier>doi:10.1101/2020.12.25.424409</dc:identifier>
<dc:title><![CDATA[Negative feedback by NUR77/Nr4a1 restrains B cell clonal dominance during early T-dependent immune responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.28.424567v1?rss=1">
<title>
<![CDATA[
eIF2B Conformation and Assembly State Regulate the Integrated Stress Response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.28.424567v1?rss=1</link>
<description><![CDATA[
The integrated stress response (ISR) is activated by phosphorylation of the translation initiation factor eIF2 in response to various stress conditions. Phosphorylated eIF2 (eIF2-P) inhibits eIF2s nucleotide exchange factor eIF2B, a two-fold symmetric heterodecamer assembled from subcomplexes. Here, we monitor and manipulate eIF2B assembly in vitro and in vivo. In the absence of eIF2Bs -subunit, the ISR is induced because unassembled eIF2B tetramer subcomplexes accumulate in cells. Upon addition of the small-molecule ISR inhibitor ISRIB, eIF2B tetramers assemble into active octamers. Surprisingly, ISRIB inhibits the ISR even in the context of fully assembled eIF2B decamers, revealing an allosteric communication between the physically distant eIF2, eIF2-P, and ISRIB binding sites. Cryo-EM structures suggest a rocking motion in eIF2B that couples these binding sites. eIF2-P binding converts eIF2B decamers into  conjoined tetramers with greatly diminished activity. Thus, ISRIBs effects in disease models could arise from eIF2B decamer stabilization, allosteric modulation, or both.
]]></description>
<dc:creator>Schoof, M.</dc:creator>
<dc:creator>Boone, M.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Lawrence, R. E.</dc:creator>
<dc:creator>Frost, A.</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:date>2020-12-29</dc:date>
<dc:identifier>doi:10.1101/2020.12.28.424567</dc:identifier>
<dc:title><![CDATA[eIF2B Conformation and Assembly State Regulate the Integrated Stress Response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.06.425642v1?rss=1">
<title>
<![CDATA[
Enhanced Solid Tumor Recognition and T cell Stemness with SynNotch CAR Circuits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.06.425642v1?rss=1</link>
<description><![CDATA[
The lack of highly tumor-specific antigens limits the development of engineered T cell therapeutics because of life-threatening "on-target/off-tumor" toxicities. Here we identify ALPPL2 as a tumor-specific antigen expressed in a spectrum of solid tumors, including mesothelioma. ALPPL2 can act as a sole target for chimeric antigen receptor (CAR) therapy or be combined with tumor-associated antigens such as MCAM or mesothelin in synthetic Notch (synNotch) CAR combinatorial antigen circuits. SynNotch CAR T cells display superior tumor control when compared to CAR T cells to the same antigens by prevention of CAR-mediated tonic signaling allowing T cells to maintain a long-lived memory and non-exhausted phenotype. Collectively, we establish ALPPL2 as a clinically viable target for multiple solid tumors and demonstrate the multi-faceted therapeutic benefits of synNotch CAR T cells.

ONE SENTENCE SUMMARYSynNotch CAR circuits targeting novel solid tumor antigens enhance specificity and improve therapeutic efficacy by regulating T cell exhaustion.
]]></description>
<dc:creator>Hyrenius-Wittsten, A.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Park, M.</dc:creator>
<dc:creator>Garcia, J. M.</dc:creator>
<dc:creator>Perry, N.</dc:creator>
<dc:creator>Montgomery, G.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:creator>Roybal, K. T.</dc:creator>
<dc:date>2021-01-07</dc:date>
<dc:identifier>doi:10.1101/2021.01.06.425642</dc:identifier>
<dc:title><![CDATA[Enhanced Solid Tumor Recognition and T cell Stemness with SynNotch CAR Circuits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.07.425754v1?rss=1">
<title>
<![CDATA[
GC-1 spg cells are most similar to Leydig cells, a testis somatic cell-type, and not germ cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.07.425754v1?rss=1</link>
<description><![CDATA[
GC-1 spg is an immortalized cell line derived from an adult mouse testis and reported to be most similar to spermatocytes, a male germ cell-type. However, immunofluorescence indicates that GC-1 spg cells express WT1, a marker of testis somatic cells, and do not express markers of germ cells. Transcriptomic profiling indicate GC-1 cells are most similar to Leydig cells. Therefore, we conclude that GC-1 spg cells are most similar to testis somatic cells.
]]></description>
<dc:creator>Norman, A. R.</dc:creator>
<dc:creator>Byrnes, L.</dc:creator>
<dc:creator>Reiter, J. R.</dc:creator>
<dc:date>2021-01-07</dc:date>
<dc:identifier>doi:10.1101/2021.01.07.425754</dc:identifier>
<dc:title><![CDATA[GC-1 spg cells are most similar to Leydig cells, a testis somatic cell-type, and not germ cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.08.425912v1?rss=1">
<title>
<![CDATA[
Caspase inhibition mitigates tau cleavage and neurotoxicity in iPSC-induced neurons with the V337M MAPT mutation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.08.425912v1?rss=1</link>
<description><![CDATA[
Tau post-translational modifications (PTMs) are associated with progressive tau accumulation and neuronal loss in tauopathies, including forms of frontotemporal lobar degeneration (FTLD) and Alzheimers disease (AD). Proteolytic cleavage of tau by active caspases, including caspase-6, represents an underexplored tau PTM implicated in tau pathology. Caspase-cleaved tau is toxic and prone to self-aggregation in experimental models. To elucidate the presence and temporal course of caspase activation, tau cleavage, and neuronal death, we generated two neoepitope monoclonal antibodies (mAbs) against caspase-6 tau proteolytic sites and cortical neurons from induced pluripotent stem cells (iPSCs) with the frontotemporal dementia (FTD)-causing V337M MAPT mutation. FTLD V337M MAPT and AD postmortem brains showed positivity for both cleaved tau mAbs as well as active caspase-6. Relative to isogenic wild-type MAPT controls, V337M MAPT neurons showed a time-dependent increase in pathogenic tau in the form of tau oligomers, caspase-cleaved tau, and p-tau. Accumulation of toxic tau species in 3-month V337M MAPT neurons also increased vulnerability to stress, which was pharmacologically rescued by caspase inhibition. We propose a model in which time-dependent accumulation of caspase-cleaved tau in V337M MAPT neurons promotes neurotoxicity that is reversed by caspase-6 inhibition. Caspase-cleaved tau may be a biomarker of tauopathy, and caspases could be viable targets for therapeutic intervention against tau pathogenesis in FTLD and other tauopathies.

SignificanceThe mechanisms leading to tau pathology in frontotemporal dementia (FTD) and Alzheimers disease (AD) remain elusive. Experimental studies in AD demonstrate that tau cleavage by active caspase-6 contributes to tau pathology since cleaved tau may be toxic and prone to self-aggregation. Yet, the role of caspase-cleaved tau in promoting toxicity and cell death is unclear. Here, we generated two neoepitope monoclonal antibodies against caspase-6 tau and evaluated tau cleavage in postmortem human brains, iPSC-induced cortical neurons with the FTD-causing V337M MAPT mutation, and isogenic wild-type MAPT controls. Our results demonstrate a time-dependent accumulation of caspase-cleaved tau and increased neurotoxicity in the mutant iNs that is reversed by caspase-6 inhibition. Caspases could be viable therapeutic targets against tau pathology in tauopathies.
]]></description>
<dc:creator>Theofilas, P.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Butler, D.</dc:creator>
<dc:creator>Morales, D. O.</dc:creator>
<dc:creator>Petersen, C.</dc:creator>
<dc:creator>Chin, B.</dc:creator>
<dc:creator>Yang, T.</dc:creator>
<dc:creator>Khan, S.</dc:creator>
<dc:creator>Ng, R.</dc:creator>
<dc:creator>Kayed, R.</dc:creator>
<dc:creator>Karch, C. M.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:creator>Temple, S.</dc:creator>
<dc:creator>Arkin, M. R.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:date>2021-01-09</dc:date>
<dc:identifier>doi:10.1101/2021.01.08.425912</dc:identifier>
<dc:title><![CDATA[Caspase inhibition mitigates tau cleavage and neurotoxicity in iPSC-induced neurons with the V337M MAPT mutation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.08.425918v1?rss=1">
<title>
<![CDATA[
A global cancer data integrator reveals principles of synthetic lethality, sex disparity and immunotherapy. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.08.425918v1?rss=1</link>
<description><![CDATA[
Advances in cancer biology are increasingly dependent on integration of heterogeneous datasets. Large scale efforts have systematically mapped many aspects of cancer cell biology; however, it remains challenging for individual scientists to effectively integrate and understand this data. We have developed a new data retrieval and indexing framework that allows us to integrate publicly available data from different sources and to combine publicly available data with new or bespoke datasets. Beyond a database search, our approach empowered testable hypotheses of new synthetic lethal gene pairs, genes associated with sex disparity, and immunotherapy targets in cancer. Our approach is straightforward to implement, well documented and is continuously updated which should enable individual users to take full advantage of efforts to map cancer cell biology.
]]></description>
<dc:creator>Yogodzinski, C. H.</dc:creator>
<dc:creator>Arab, A.</dc:creator>
<dc:creator>Pritchard, J. R.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Gilbert, L.</dc:creator>
<dc:date>2021-01-09</dc:date>
<dc:identifier>doi:10.1101/2021.01.08.425918</dc:identifier>
<dc:title><![CDATA[A global cancer data integrator reveals principles of synthetic lethality, sex disparity and immunotherapy.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.11.426198v1?rss=1">
<title>
<![CDATA[
Experimental and mathematical insights on the interactions between poliovirus and a defective interfering genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.11.426198v1?rss=1</link>
<description><![CDATA[
1During replication, RNA viruses accumulate genome alterations, such as mutations and deletions. The interactions between individual variants can determine the fitness of the virus population and, thus, the outcome of infection. To investigate the effects of defective interfering genomes (DI) on wild-type (WT) poliovirus replication, we developed an ordinary differential equation model. We experimentally determined virus and DI replication during co-infection, and use these data to infer model parameters. Our model predicts, and our experimental measurements confirm, that DI replication and encapsidation are the most important determinants for the outcome of the competition. WT replication inversely correlates with DI replication. Our model predicts that genome replication and effective DI genome encapsidation are critical to effectively inhibit WT production, but an equilibrium can be established which enables WT to replicate, albeit to reduce levels.
]]></description>
<dc:creator>Andino, R.</dc:creator>
<dc:creator>Shirogane, Y.</dc:creator>
<dc:creator>Rousseau, E.</dc:creator>
<dc:creator>Voznica, J.</dc:creator>
<dc:creator>Xiao, Y.</dc:creator>
<dc:creator>Su, W.</dc:creator>
<dc:creator>Catching, A.</dc:creator>
<dc:creator>Whitfiled, Z. J.</dc:creator>
<dc:creator>Rouzine, I. M.</dc:creator>
<dc:creator>Bianco, S.</dc:creator>
<dc:date>2021-01-11</dc:date>
<dc:identifier>doi:10.1101/2021.01.11.426198</dc:identifier>
<dc:title><![CDATA[Experimental and mathematical insights on the interactions between poliovirus and a defective interfering genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.13.426598v1?rss=1">
<title>
<![CDATA[
De novo protein fold families expand the designable ligand binding site space 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.13.426598v1?rss=1</link>
<description><![CDATA[
A major challenge in designing proteins de novo to bind user-defined ligands with high specificity and affinity is finding backbones structures that can accommodate a desired binding site geometry with high precision. Recent advances in methods to generate protein fold families de novo have expanded the space of accessible protein structures, but it is not clear to what extend de novo proteins with diverse geometries also expand the space of designable ligand binding functions. We constructed a library of 25,806 high-quality ligand binding sites and developed a fast protocol to place ("match") these binding sites into both naturally occurring and de novo protein families with two fold topologies: Rossman and NTF2. 5,896 and 7,475 binding sites could be matched to the Rossmann and NTF2 fold families, respectively. De novo designed Rossman and NTF2 protein families can support 1,791 and 678 binding sites that cannot be matched to naturally existing structures with the same topologies, respectively. While the number of protein residues in ligand binding sites is the major determinant of matching success, ligand size and primary sequence separation of binding site residues also play important roles. The number of matched binding sites are power law functions of the number of members in a fold family. Our results suggest that de novo sampling of geometric variations on diverse fold topologies can significantly expand the space of designable ligand binding sites for a wealth of possible new protein functions.

Author summaryDe novo design of proteins that can bind to novel and highly diverse user-defined small molecule ligands could have broad biomedical and synthetic biology applications. Because ligand binding site geometries need to be accommodated by protein backbone scaffolds at high accuracy, the diversity of scaffolds is a major limitation for designing new ligand binding functions. Advances in computational protein structure design methods have significantly increased the number of accessible stable scaffold structures. Understanding how many new ligand binding sites can be accommodated by the de novo scaffolds is important for designing novel ligand binding proteins. To answer this question, we constructed a large library of ligand binding sites from the Protein Data Bank (PDB). We tested the number of ligand binding sites that can be accommodated by de novo scaffolds and naturally existing scaffolds with same fold topologies. The results showed that de novo scaffolds significantly expanded the ligand binding space of their respective fold topologies. We also identified factors that affect difficulties of binding site accommodation, as well as the relationship between the number of scaffolds and the accessible ligand binding site space. We believe our findings will benefit future method development and applications of ligand binding protein design.
]]></description>
<dc:creator>Pan, X.</dc:creator>
<dc:creator>Kortemme, T.</dc:creator>
<dc:date>2021-01-15</dc:date>
<dc:identifier>doi:10.1101/2021.01.13.426598</dc:identifier>
<dc:title><![CDATA[De novo protein fold families expand the designable ligand binding site space]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.17.427024v1?rss=1">
<title>
<![CDATA[
Tropism of SARS-CoV-2 for Developing Human Cortical Astrocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.17.427024v1?rss=1</link>
<description><![CDATA[
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) readily infects a variety of cell types impacting the function of vital organ systems, with particularly severe impact on respiratory function. It proves fatal for one percent of those infected. Neurological symptoms, which range in severity, accompany a significant proportion of COVID-19 cases, indicating a potential vulnerability of neural cell types. To assess whether human cortical cells can be directly infected by SARS-CoV-2, we utilized primary human cortical tissue and stem cell-derived cortical organoids. We find significant and predominant infection in cortical astrocytes in both primary and organoid cultures, with minimal infection of other cortical populations. Infected astrocytes had a corresponding increase in reactivity characteristics, growth factor signaling, and cellular stress. Although human cortical cells, including astrocytes, have minimal ACE2 expression, we find high levels of alternative coronavirus receptors in infected astrocytes, including DPP4 and CD147. Inhibition of DPP4 reduced infection and decreased expression of the cell stress marker, ARCN1. We find tropism of SARS-CoV-2 for human astrocytes mediated by DPP4, resulting in reactive gliosis-type injury.
]]></description>
<dc:creator>Andrews, M. G.</dc:creator>
<dc:creator>Mukhtar, T.</dc:creator>
<dc:creator>Eze, U. C.</dc:creator>
<dc:creator>Simoneau, C. R.</dc:creator>
<dc:creator>Perez, Y.</dc:creator>
<dc:creator>Mostajo-Radji, M. A.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Velmeshev, D.</dc:creator>
<dc:creator>Salma, J.</dc:creator>
<dc:creator>Kumar, G. R.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:creator>Crouch, E. E.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Kriegstein, A. R.</dc:creator>
<dc:date>2021-01-18</dc:date>
<dc:identifier>doi:10.1101/2021.01.17.427024</dc:identifier>
<dc:title><![CDATA[Tropism of SARS-CoV-2 for Developing Human Cortical Astrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.18.427101v1?rss=1">
<title>
<![CDATA[
Treponema denticola dentilisin triggered TLR2/MyD88 activation upregulates a tissue destructive program involving MMPs via Sp1 in human oral cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.18.427101v1?rss=1</link>
<description><![CDATA[
Periodontal disease is driven by dysbiosis of the oral microbiome, resulting in over-representation of species that induce the release of pro-inflammatory cytokines, chemokines, and tissue-remodeling matrix metalloproteinases (MMPs) in the periodontium. These chronic tissue-destructive inflammatory responses result in gradual loss of tooth-supporting alveolar bone. The oral spirochete Treponema denticola, is consistently found at significantly elevated levels in periodontal lesions. Host-expressed Toll-Like Receptor 2 (TLR2) senses a variety of bacterial ligands, including acylated lipopolysaccharides and lipoproteins. T. denticola dentilisin, a surface-expressed protease complex comprised of three lipoproteins has been implicated as a virulence factor in periodontal disease, primarily due to its proteolytic activity. While the role of acylated bacterial components in induction of inflammation is well-studied, little attention has been given to the potential role of the acylated nature of dentilisin. The purpose of this study was to test the hypothesis that T. denticola dentilisin activates a TLR2-dependent mechanism, leading to upregulation of tissue-destructive genes in periodontal tissue. RNA-sequencing of periodontal ligament cells challenged with T. denticola bacteria revealed a significant upregulation of genes associated with extracellular matrix organization and degradation, including tissue-specific inducible MMPs that may play novel roles in modulating host immune responses yet to be characterized within the context of oral disease. The Gram-negative oral commensal, Veillonella parvula, failed to upregulate these same MMPs. Dentilisin-induced upregulation of MMPs was mediated via TLR2 and MyD88 activation, since knockdown of either TLR2 or MyD88 abrogated these effects. Challenge with purified dentilisin upregulated the same MMPs, whereas a dentilisin-deficient T. denticola mutant had no effect. Finally, T. denticola-mediated activation of TLR2/MyD88 led to the nuclear translocation of the transcription factor Sp1, which was shown to be a critical regulator of all T. denticola-dependent MMP expression. Taken together, these data support that T. denticola dentilisin stimulates tissue-destructive cellular processes in a TLR2/MyD88/Sp1-dependent fashion.

AUTHOR SUMMARYPeriodontal disease is driven by dysbiosis of the oral microbiome, which interacts with host tissues and thereby induces the release of pro-inflammatory cytokines, chemokines, and tissue-remodeling matrix metalloproteinases (MMPs), leading to destruction of the periodontal tissues. Even after clinical intervention, patients with severe periodontal disease are left with a persistent pro-inflammatory transcriptional profile throughout the periodontium. The oral spirochete, Treponema denticola, is consistently found at elevated levels in periodontal lesions and is associated with several pathophysiological effects driving periodontal disease progression. The T. denticola surface-expressed protease complex (dentilisin) has cytopathic effects consistent with periodontal disease pathogenesis. To date, few direct links have been reported between dentilisin and the cellular and tissue processes that drive periodontal tissue destruction at the transcriptional and/or epigenetic levels. Here, we utilize wild type and dentilisin-deficient T. denticola as well as purified dentilisin to characterize dentilisin-dependent activation of intracellular pathways controlling MMP expression and activity. Our results define a role for dentilisin in initiating this signal cascade. Also, our study identified tissue-specific inducible MMPs that may play novel roles in modulating as-yet uncharacterized host responses in periodontal disease. Lastly, T. denticola dentilisin stimulates tissue-destructive cellular processes in a TLR2/MyD88/Sp1-dependent fashion. Taken together, our study provides new insights into the molecular mechanisms underpinning periodontal disease progression which could lead to the development of more efficacious therapeutic treatments.
]]></description>
<dc:creator>Ganther, S.</dc:creator>
<dc:creator>Radaic, A.</dc:creator>
<dc:creator>Chang, N.-Y. N.</dc:creator>
<dc:creator>Tafolla, C.</dc:creator>
<dc:creator>Zhan, L.</dc:creator>
<dc:creator>Fenno, J. C.</dc:creator>
<dc:creator>Kapila, Y.</dc:creator>
<dc:date>2021-01-18</dc:date>
<dc:identifier>doi:10.1101/2021.01.18.427101</dc:identifier>
<dc:title><![CDATA[Treponema denticola dentilisin triggered TLR2/MyD88 activation upregulates a tissue destructive program involving MMPs via Sp1 in human oral cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.18.427195v1?rss=1">
<title>
<![CDATA[
Molecular programs of fibrotic change in aging human lung 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.18.427195v1?rss=1</link>
<description><![CDATA[
Aging is associated with both overt and subclinical lung fibrosis, which increases risk for mortality from viruses and other respiratory pathogens. The molecular programs that induce fibrosis in the aging lung are not well understood. To overcome this knowledge gap, we undertook multimodal profiling of distal lung samples from healthy human donors across the lifespan. Telomere shortening, a cause of cell senescence and fibrosis, was progressive with age in a sample of 86 lungs and was associated with foci of DNA damage. Bulk RNA sequencing confirmed activation of cellular senescence and pro-fibrotic pathways as well as genes necessary for collagen processing with increasing age. These findings were validated in independent datasets for lung and sun-exposed skin, but not other organs including heart, liver and kidney. Cell type deconvolution analysis revealed a progressive loss of lung epithelial cells and an increasing proportion of fibroblasts. Consistent with the observed pro-fibrotic transcriptional profile, second harmonic imaging demonstrated increased density of interstitial collagen in aged human lungs. Furthermore, regions of parenchymal fibrosis were associated with decreased alveolar expansion and surfactant secretion. These findings reveal the transcriptional and structural features of fibrosis and associated physiologic impairments in normal lung aging.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Islam, M. N.</dc:creator>
<dc:creator>Boostanpour, K.</dc:creator>
<dc:creator>Aran, D.</dc:creator>
<dc:creator>Christenson, S.</dc:creator>
<dc:creator>Matthay, M. A.</dc:creator>
<dc:creator>Eckalbar, W.</dc:creator>
<dc:creator>DePianto, D. J.</dc:creator>
<dc:creator>Arron, J. R.</dc:creator>
<dc:creator>Magee, L.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Matsumoto, R.</dc:creator>
<dc:creator>Kubota, M.</dc:creator>
<dc:creator>Farber, D. L.</dc:creator>
<dc:creator>Bhattacharya, J.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:creator>Bhattacharya, M.</dc:creator>
<dc:date>2021-01-19</dc:date>
<dc:identifier>doi:10.1101/2021.01.18.427195</dc:identifier>
<dc:title><![CDATA[Molecular programs of fibrotic change in aging human lung]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.26.428277v1?rss=1">
<title>
<![CDATA[
Mechanotaxis directs Pseudomonas aeruginosa twitching motility. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.26.428277v1?rss=1</link>
<description><![CDATA[
The opportunistic pathogen Pseudomonas aeruginosa explores surfaces using twitching motility powered by retractile extracellular filaments called type IV pili. Single cells twitch by successive pili extension, attachment and retraction. However, whether and how single cells control twitching migration remains unclear. We discovered that P. aeruginosa actively directs twitching in the direction of mechanical input from type IV pili, in a process we call mechanotaxis. The Chp chemotaxis-like system controls the balance of forward and reverse twitching migration of single cells in response to the mechanical signal. On surfaces, Chp senses type IV pili attachment at one pole thereby sensing a spatially-resolved signal. As a result, the Chp response regulators PilG and PilH control the polarization of the extension motor PilB. PilG stimulates polarization favoring forward migration, while PilH inhibits polarization inducing reversal. Subcellular segregation of PilG and PilH efficiently orchestrates their antagonistic functions, ultimately enabling rapid reversals upon perturbations. This distinct localization of response regulators establishes a signaling landscape known as local-excitation, global-inhibition in higher order organisms, identifying a conserved strategy to transduce spatially-resolved signals. Our discovery finally resolves the function of the Chp system and expands our view of the signals regulating motility.
]]></description>
<dc:creator>Tala, L.</dc:creator>
<dc:creator>Kuhn, M.</dc:creator>
<dc:creator>Inclan, Y.</dc:creator>
<dc:creator>Patino, R.</dc:creator>
<dc:creator>Pierrat, X.</dc:creator>
<dc:creator>Vos, I.</dc:creator>
<dc:creator>Al-Mayyah, Z.</dc:creator>
<dc:creator>MacMillan, H.</dc:creator>
<dc:creator>Negrete, J.</dc:creator>
<dc:creator>Engel, J. N.</dc:creator>
<dc:creator>Persat, A.</dc:creator>
<dc:date>2021-01-26</dc:date>
<dc:identifier>doi:10.1101/2021.01.26.428277</dc:identifier>
<dc:title><![CDATA[Mechanotaxis directs Pseudomonas aeruginosa twitching motility.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.27.428524v1?rss=1">
<title>
<![CDATA[
A Human Accelerated Region is a Leydig cell GLI2 Enhancer that Affects Male-Typical Behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.27.428524v1?rss=1</link>
<description><![CDATA[
Human accelerated regions (HARs) are sequences that have evolved at an accelerated rate in the human lineage. Some HARs are developmental enhancers. We used a massively parallel reporter assay (MPRA) to identify HARs with enhancer activity in a mammalian testis cell line. A subset of HARs exhibited differential activity between the human and chimpanzee orthologs, representing candidates for underlying unique human male reproductive biology. We further characterized one of these candidate testis enhancers, 2xHAR.238. CRISPR/Cas9-mediated deletion in a testis cell line and mice revealed that 2xHAR.238 enhances expression of Gli2, encoding a Hedgehog pathway effector, in testis Leydig cells. 4C-seq revealed that 2xHAR.238 contacts the Gli2 promoter, consistent with enhancer function. In adult male mice, deletion of 2xHAR.238 disrupted mouse male-typical behavior and male interest in female odor. Combined, our work identifies a HAR that promotes the expression of Gli2 in Leydig cells and may have contributed to the evolution of human male reproductive biology.
]]></description>
<dc:creator>Reiter, J.</dc:creator>
<dc:creator>Norman, A. R.</dc:creator>
<dc:creator>Ahituv, N. R.</dc:creator>
<dc:creator>Ryu, A. H.</dc:creator>
<dc:creator>Jamieson, K.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Thomas, S.</dc:creator>
<dc:date>2021-01-27</dc:date>
<dc:identifier>doi:10.1101/2021.01.27.428524</dc:identifier>
<dc:title><![CDATA[A Human Accelerated Region is a Leydig cell GLI2 Enhancer that Affects Male-Typical Behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.29.428822v1?rss=1">
<title>
<![CDATA[
NR2F1 is a barrier to dissemination of early-evolved mammary cancer cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.29.428822v1?rss=1</link>
<description><![CDATA[
SO_SCPLOWUMMARYC_SCPLOWCancer cells disseminate during very early and sometimes asymptomatic stages of tumor progression. Granted that biological barriers to tumorigenesis exist, there must also be limiting steps to early dissemination, all of which remain largely unknown. We report that the orphan nuclear receptor NR2F1/COUP-TF1 serves as a barrier to early dissemination. High-resolution intravital imaging revealed that loss of function of NR2F1 in HER2+ early cancer cells increased in vivo dissemination without accelerating mammary tumor formation. NR2F1 expression was positively regulated by the tumor suppressive MMK3/6-p38-MAPK pathway and downregulated by HER2 and Wnt4 oncogenic signaling. NR2F1 downregulation by HER2 in early cancer cells led to decreased E-cadherin expression and {beta}-catenin membrane localization, disorganized laminin 5 deposition, and increased expression of CK14, TWIST1, ZEB1 and PRRX1. Our findings reveal the existence of an inhibitory mechanism of dissemination regulated by NR2F1 downstream of HER2 signaling.
]]></description>
<dc:creator>Rodriguez Tirado, C. A.</dc:creator>
<dc:creator>Kale, N.</dc:creator>
<dc:creator>Carlini, M. J.</dc:creator>
<dc:creator>Shrivastava, N.</dc:creator>
<dc:creator>Khalil, B. D.</dc:creator>
<dc:creator>Bravo-Cordero, J. J.</dc:creator>
<dc:creator>Alexander, M.</dc:creator>
<dc:creator>Ji, J.</dc:creator>
<dc:creator>Sosa, M. S.</dc:creator>
<dc:date>2021-01-29</dc:date>
<dc:identifier>doi:10.1101/2021.01.29.428822</dc:identifier>
<dc:title><![CDATA[NR2F1 is a barrier to dissemination of early-evolved mammary cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.02.425931v1?rss=1">
<title>
<![CDATA[
Cofilin pathology is a new player on α-synuclein-induced spine impairment in models of hippocampal synucleinopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.02.425931v1?rss=1</link>
<description><![CDATA[
Cognitive dysfunction and dementia are presently recognized as major complications in -synucleinopathies, namely in Dementia with Lewy Bodies (DLB) and Parkinsons disease with dementia (PDD). In these disorders, -Synuclein (Syn) accumulation affects severely the hippocampus by inducing synaptic dysfunction which culminates in cognitive impairment. To characterize the mechanisms underlying Syn-induced neuronal dysfunction we analysed the effect of overexpression or extracellular administration of Syn on hippocampal neurons. We observed that Syn induces the dysregulation of the actin-binding protein cofilin and its assembly into rod structures in a mechanism mediated by the cellular prion protein (PrPC). Moreover, we unraveled cofilin pathology as mediator of Syn-induced dendritic spine impairment in hippocampal neurons. Importantly, in a synucleinopathy mouse model with cognitive impairment we validated cofilin dysregulation and synaptic dysfunction at the same age when cognitive deficits were observed. Our data supports cofilin as a novel player on hippocampal synaptic dysfunction triggered by Syn on Lewy Body dementias.
]]></description>
<dc:creator>Oliveira da Silva, M. I.</dc:creator>
<dc:creator>Santejo, M.</dc:creator>
<dc:creator>Babcock, I. W.</dc:creator>
<dc:creator>Magalhaes, A.</dc:creator>
<dc:creator>Minamide, L. S.</dc:creator>
<dc:creator>Castillo, E.</dc:creator>
<dc:creator>Gerhardt, E.</dc:creator>
<dc:creator>Fahlbusch, C.</dc:creator>
<dc:creator>Swanson, R. A.</dc:creator>
<dc:creator>Outeiro, T. F.</dc:creator>
<dc:creator>Bamburg, J. R.</dc:creator>
<dc:creator>Liz, M. A.</dc:creator>
<dc:date>2021-02-02</dc:date>
<dc:identifier>doi:10.1101/2021.02.02.425931</dc:identifier>
<dc:title><![CDATA[Cofilin pathology is a new player on α-synuclein-induced spine impairment in models of hippocampal synucleinopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.02.429276v1?rss=1">
<title>
<![CDATA[
Deep Proteome Profiling of Human Mammary Epithelia at Lineage and Age Resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.02.429276v1?rss=1</link>
<description><![CDATA[
Age is the major risk factor in most carcinomas, yet little is known about how proteomes change with age in any human epithelium. We present comprehensive proteomes comprised of >9,000 total proteins, and >15,000 phosphopeptides, from normal primary human mammary epithelia at lineage resolution from ten women ranging in age from 19 to 68. Data were quality controlled, and results were biologically validated with cell-based assays. Age-dependent protein signatures were identified using differential expression analyses and weighted protein co-expression network analyses. Up-regulation of basal markers in luminal cells, including KRT14 and AXL, were a prominent consequence of aging. PEAK1 was identified as an age-dependent signaling kinase in luminal cells, which revealed a potential age-dependent vulnerability for targeted ablation. Correlation analyses between transcriptome and proteome revealed age-associated loss of proteostasis regulation. Protein expression and phosphorylation changes in the aging breast epithelium identify potential therapeutic targets for reducing breast cancer susceptibility.
]]></description>
<dc:creator>Hinz, S.</dc:creator>
<dc:creator>Manousopoulou, A.</dc:creator>
<dc:creator>Miyano, M.</dc:creator>
<dc:creator>Sayaman, R. W.</dc:creator>
<dc:creator>Aguilera, K. Y.</dc:creator>
<dc:creator>Todhunter, M. E.</dc:creator>
<dc:creator>Lopez, J. C.</dc:creator>
<dc:creator>Sohn, L. L.</dc:creator>
<dc:creator>Wang, L. D.</dc:creator>
<dc:creator>LaBarge, M. A.</dc:creator>
<dc:date>2021-02-02</dc:date>
<dc:identifier>doi:10.1101/2021.02.02.429276</dc:identifier>
<dc:title><![CDATA[Deep Proteome Profiling of Human Mammary Epithelia at Lineage and Age Resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.02.429423v1?rss=1">
<title>
<![CDATA[
Paradoxical hyperexcitability from NaV1.2 sodium channel loss in neocortical pyramidal cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.02.429423v1?rss=1</link>
<description><![CDATA[
Loss-of-function variants in the gene SCN2A, which encodes the sodium channel NaV1.2, are strongly associated with autism spectrum disorder and intellectual disability. An estimated 20-30% of children with these variants are co-morbid for epilepsy, with altered neuronal activity originating in neocortex, a region where NaV1.2 channels are expressed predominantly in excitatory pyramidal cells. This is paradoxical, as sodium channel loss in excitatory cells would be expected to dampen neocortical activity rather than promote seizure. Here, we examined pyramidal neurons lacking NaV1.2 channels and found that they were intrinsically hyperexcitable, firing high-frequency bursts of action potentials (APs) despite decrements in AP size and speed. Compartmental modeling and dynamic clamp recordings revealed that NaV1.2 loss prevented potassium channels from properly repolarizing neurons between APs, increasing overall excitability by allowing neurons to reach threshold for subsequent APs more rapidly. This cell-intrinsic mechanism may therefore account for why SCN2A loss-of-function can paradoxically promote seizure.
]]></description>
<dc:creator>Spratt, P. W.</dc:creator>
<dc:creator>Ben-Shalom, R.</dc:creator>
<dc:creator>Sahagun, A.</dc:creator>
<dc:creator>Keeshen, C. M.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Bender, K. J.</dc:creator>
<dc:date>2021-02-02</dc:date>
<dc:identifier>doi:10.1101/2021.02.02.429423</dc:identifier>
<dc:title><![CDATA[Paradoxical hyperexcitability from NaV1.2 sodium channel loss in neocortical pyramidal cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.09.430269v1?rss=1">
<title>
<![CDATA[
Integrated plasma proteomic and single-cell immune signaling network signatures demarcate mild, moderate, and severe COVID-19 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.09.430269v1?rss=1</link>
<description><![CDATA[
The biological determinants of the wide spectrum of COVID-19 clinical manifestations are not fully understood. Here, over 1400 plasma proteins and 2600 single-cell immune features comprising cell phenotype, basal signaling activity, and signaling responses to inflammatory ligands were assessed in peripheral blood from patients with mild, moderate, and severe COVID-19, at the time of diagnosis. Using an integrated computational approach to analyze the combined plasma and single-cell proteomic data, we identified and independently validated a multivariate model classifying COVID-19 severity (multi-class AUCtraining = 0.799, p-value = 4.2e-6; multi-class AUCvalidation = 0.773, p-value = 7.7e-6). Features of this high-dimensional model recapitulated recent COVID-19 related observations of immune perturbations, and revealed novel biological signatures of severity, including the mobilization of elements of the renin-angiotensin system and primary hemostasis, as well as dysregulation of JAK/STAT, MAPK/mTOR, and NF-{kappa}B immune signaling networks. These results provide a set of early determinants of COVID-19 severity that may point to therapeutic targets for the prevention of COVID-19 progression.

SummaryFeyaerts et al. demonstrate that an integrated analysis of plasma and single-cell proteomics differentiates COVID-19 severity and reveals severity-specific biological signatures associated with the dysregulation of the JAK/STAT, MAPK/mTOR, and NF-{kappa}B immune signaling networks and the mobilization of the renin-angiotensin and hemostasis systems.
]]></description>
<dc:creator>Feyaerts, D.</dc:creator>
<dc:creator>Hedou, J.</dc:creator>
<dc:creator>Gillard, J.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Tsai, E. S.</dc:creator>
<dc:creator>Peterson, L. S.</dc:creator>
<dc:creator>Ando, K.</dc:creator>
<dc:creator>Manohar, M.</dc:creator>
<dc:creator>Do, E.</dc:creator>
<dc:creator>Dhondalay, G. K. R.</dc:creator>
<dc:creator>Fitzpatrick, J.</dc:creator>
<dc:creator>Artandi, M.</dc:creator>
<dc:creator>Chang, I.</dc:creator>
<dc:creator>Snow, T. T.</dc:creator>
<dc:creator>Chinthrajah, R. S.</dc:creator>
<dc:creator>Warren, C. M.</dc:creator>
<dc:creator>Wittman, R.</dc:creator>
<dc:creator>Meyerowitz, J. G.</dc:creator>
<dc:creator>Ganio, E. A.</dc:creator>
<dc:creator>Stelzer, I. A.</dc:creator>
<dc:creator>Han, X.</dc:creator>
<dc:creator>Verdonk, F.</dc:creator>
<dc:creator>Gaudilliere, D. K.</dc:creator>
<dc:creator>Mukherjee, N.</dc:creator>
<dc:creator>Tsai, A. S.</dc:creator>
<dc:creator>Rumer, K. K.</dc:creator>
<dc:creator>Jiang, S. J.</dc:creator>
<dc:creator>Valdes-Ferrer, S. I.</dc:creator>
<dc:creator>Kelly, J. D.</dc:creator>
<dc:creator>Furman, D.</dc:creator>
<dc:creator>Aghaeepour, N.</dc:creator>
<dc:creator>Angst, M. S.</dc:creator>
<dc:creator>Boyd, S. D.</dc:creator>
<dc:creator>Pinsky, B. A.</dc:creator>
<dc:creator>Nolan, G. P.</dc:creator>
<dc:creator>Nadeau, K. C.</dc:creator>
<dc:creator>Gaudilliere, B.</dc:creator>
<dc:creator>McIlwain, D. R.</dc:creator>
<dc:date>2021-02-12</dc:date>
<dc:identifier>doi:10.1101/2021.02.09.430269</dc:identifier>
<dc:title><![CDATA[Integrated plasma proteomic and single-cell immune signaling network signatures demarcate mild, moderate, and severe COVID-19]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.09.430368v1?rss=1">
<title>
<![CDATA[
Identifying the genetic basis and molecular mechanisms underlying phenotypic correlation between complex human traits using a gene-based approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.09.430368v1?rss=1</link>
<description><![CDATA[
Phenotypic correlations between complex human traits have long been observed based on epidemiological studies. However, the genetic basis and underlying mechanisms are largely unknown. Here we developed a gene-based approach to measure genetic overlap between a pair of traits and to delineate the shared genes/pathways, through three steps: 1) translating SNP-phenotype association profile to gene-phenotype association profile by integrating GWAS with eQTL data using a newly developed algorithm called Sherlock-II; 2) measuring the genetic overlap between a pair of traits by a normalized distance and the associated p value between the two gene-phenotype association profiles; 3) delineating genes/pathways involved. Application of this approach to a set of GWAS data covering 59 human traits detected significant overlap between many known and unexpected pairs of traits; a significant fraction of them are not detectable by SNP based genetic similarity measures. Examples include Cancer and Alzheimers Disease (AD), Rheumatoid Arthritis and Crohns disease, and Longevity and Fasting glucose. Functional analysis revealed specific genes/pathways shared by these pairs. For example, Cancer and AD are co-associated with genes involved in hypoxia response and P53/apoptosis pathways, suggesting specific mechanisms underlying the inverse correlation between them. Our approach can detect yet unknown relationships between complex traits and generate mechanistic hypotheses and has the potential to improve diagnosis and treatment by transferring knowledge from one disease to another.
]]></description>
<dc:creator>Gu, J.</dc:creator>
<dc:creator>Fuller, C. K.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2021-02-10</dc:date>
<dc:identifier>doi:10.1101/2021.02.09.430368</dc:identifier>
<dc:title><![CDATA[Identifying the genetic basis and molecular mechanisms underlying phenotypic correlation between complex human traits using a gene-based approach]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.11.430844v1?rss=1">
<title>
<![CDATA[
Phenotypic analysis of catastrophic childhood epilepsy genes: The Epilepsy Zebrafish Project 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.11.430844v1?rss=1</link>
<description><![CDATA[
Genetic engineering techniques have contributed to the now widespread use of zebrafish to investigate gene function, but zebrafish-based human disease studies, and particularly for neurological disorders, are limited. Here we used CRISPR-Cas9 to generate 40 single-gene mutant zebrafish lines representing catastrophic childhood epilepsies. We evaluated larval phenotypes using electrophysiological, behavioral, neuro-anatomical, survival and pharmacological assays. Phenotypes with unprovoked electrographic seizure activity (i.e., epilepsy) were identified in zebrafish lines for 8 genes; ARX, EEF1A, GABRB3, GRIN1, PNPO, SCN1A, STRADA and STXBP1. A unifying epilepsy classification scheme was developed based on local field potential recordings and blinded scoring from ~3300 larvae. We also created an open-source database containing sequencing information, survival curves, behavioral profiles and representative electrophysiology data. We offer all zebrafish lines as a resource to the neuroscience community and envision them as a starting point for further functional analysis and/or identification of new therapies.
]]></description>
<dc:creator>Griffin, A.</dc:creator>
<dc:creator>Carpenter, C.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Grone, B.</dc:creator>
<dc:creator>Hamling, K.</dc:creator>
<dc:creator>Moog, M.</dc:creator>
<dc:creator>Dinday, M. T.</dc:creator>
<dc:creator>Figueroa, F.</dc:creator>
<dc:creator>Anvar, M.</dc:creator>
<dc:creator>Ononuju, C.</dc:creator>
<dc:creator>Qu, T.</dc:creator>
<dc:creator>Baraban, S. C.</dc:creator>
<dc:date>2021-02-12</dc:date>
<dc:identifier>doi:10.1101/2021.02.11.430844</dc:identifier>
<dc:title><![CDATA[Phenotypic analysis of catastrophic childhood epilepsy genes: The Epilepsy Zebrafish Project]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.14.431158v1?rss=1">
<title>
<![CDATA[
Multisite phosphorylation by Cdk1 initiates delayed negative feedback to control mitotic transcription 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.14.431158v1?rss=1</link>
<description><![CDATA[
Cell-cycle progression is driven by the phosphorylation of cyclin-dependent kinase (Cdk) substrates1-3. The order of substrate phosphorylation depends in part on the general rise in Cdk activity during the cell cycle4-7, together with variations in substrate docking to sites on associated cyclin and Cks subunits3, 6, 8-10. Many substrates are modified at multiple sites to provide more complex regulation9, 11-14. Here, we describe an elegant regulatory circuit based on multisite phosphorylation of Ndd1, a transcriptional co-activator of genes required for mitotic progression15, 16. As cells enter mitosis, Ndd1 phosphorylation by Cdk1 is known to promote mitotic cyclin (CLB2) gene transcription, resulting in positive feedback17-20. Consistent with these findings, we show that low Cdk1 activity promotes CLB2 expression at mitotic entry. We also find, however, that CLB2 expression is inhibited by high levels of Cdk1 activity in a mitotic arrest. Inhibition is accompanied by Ndd1 degradation, and we present evidence that high mitotic Cdk1-Clb2 activity generates phosphodegrons on Ndd1, leading to its degradation. Complete Ndd1 phosphorylation by the Clb2-Cdk1-Cks1 complex requires the phosphothreonine-binding site of Cks1, as well as a novel phosphate-binding pocket on the cyclin Clb221. We therefore propose that initial phosphorylation by Cdk1 primes the protein for secondary phosphorylation at phosphodegrons, resulting in degradation only at high Cdk1 activity. Together, our results suggest that rising levels of mitotic Cdk1 activity act at multiple phosphorylation sites on Ndd1, first triggering rapid positive feedback and then promoting delayed negative feedback, resulting in a pulse of mitotic gene expression.
]]></description>
<dc:creator>Asfaha, J.</dc:creator>
<dc:creator>Ord, M.</dc:creator>
<dc:creator>Loog, M.</dc:creator>
<dc:creator>Morgan, D. O.</dc:creator>
<dc:date>2021-02-14</dc:date>
<dc:identifier>doi:10.1101/2021.02.14.431158</dc:identifier>
<dc:title><![CDATA[Multisite phosphorylation by Cdk1 initiates delayed negative feedback to control mitotic transcription]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.18.431915v1?rss=1">
<title>
<![CDATA[
Glioblastoma remodeling of neural circuits in the human brain decreases survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.18.431915v1?rss=1</link>
<description><![CDATA[
Gliomas synaptically integrate into neural circuits. Prior work has demonstrated bidirectional interactions between neurons and glioma cells, with neuronal activity driving glioma growth and gliomas increasing neuronal excitability. In this study we wanted to know how glioma induced neuronal changes influence neural circuits underlying cognition and whether these interactions influence patient survival. We use intracranial brain recordings during lexical retrieval language tasks in awake humans in addition to site specific tumor tissue biopsies and cell biology experiments. We find that gliomas remodel functional neural circuitry such that task-relevant neural responses activate tumor-infiltrated cortex, beyond cortical excitation normally recruited in the healthy brain. Site-directed biopsies from functionally connected regions within the tumor are enriched for a glioblastoma subpopulation that exhibits a distinct synaptogenic and neuronotrophic phenotype. Tumor cells from functionally connected regions secrete the synaptogenic factor thrombospondin-1, which contributes to the differential neuron-glioma interactions observed in functionally connected tumor regions compared to tumor regions with less functional connectivity. The degree of functional connectivity between glioblastoma and the normal brain negatively impacts both patient survival and language task performance. These data demonstrate that high-grade gliomas functionally remodel neural circuits in the human brain, which both promotes tumor proliferation and impairs cognition.
]]></description>
<dc:creator>Krishna, S.</dc:creator>
<dc:creator>Choudhury, A.</dc:creator>
<dc:creator>Seo, K.</dc:creator>
<dc:creator>Ni, L.</dc:creator>
<dc:creator>Kakaizada, S.</dc:creator>
<dc:creator>Lee, A.</dc:creator>
<dc:creator>Aabedi, A.</dc:creator>
<dc:creator>Cao, C.</dc:creator>
<dc:creator>Sudharshan, R.</dc:creator>
<dc:creator>Egladyous, A.</dc:creator>
<dc:creator>Almeida, N.</dc:creator>
<dc:creator>Venkatesh, H.</dc:creator>
<dc:creator>Findlay, A.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Raleigh, D.</dc:creator>
<dc:creator>Brang, D.</dc:creator>
<dc:creator>Monje, M.</dc:creator>
<dc:creator>Hervey-Jumper, S.</dc:creator>
<dc:date>2021-02-19</dc:date>
<dc:identifier>doi:10.1101/2021.02.18.431915</dc:identifier>
<dc:title><![CDATA[Glioblastoma remodeling of neural circuits in the human brain decreases survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.22.432272v1?rss=1">
<title>
<![CDATA[
Microglial NF-κB drives tau spreading and toxicity in a mouse model of tauopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.22.432272v1?rss=1</link>
<description><![CDATA[
Activation of microglia, the brains innate immune cells, is a prominent pathological feature in tauopathies, including Alzheimers disease. How microglia activation contributes to tau toxicity remains largely unknown. Here we show that nuclear factor kappa-light-chain-enhancer of activated B cells (NF-{kappa}B) signaling, activated by tau, drives microglial-mediated tau propagation and toxicity. Constitutive activation of microglial NF-{kappa}B exacerbated, while inactivation diminished, tau seeding and spreading in PS19 mice, consistent with the observation that NF-{kappa}B activation accelerates processing of internalized tau fibrils in primary microglia. Remarkably, inhibition of microglial NF-{kappa}B specifically also rescued tau-mediated learning and memory deficits, and restored overall transcriptomic changes while increasing tau inclusions. On a single cell level, we discovered that tau-associated disease states in microglia were diminished by NF-{kappa}B inactivation and further transformed by constitutive NF-{kappa}B activation. Our study establishes a central role for microglial NF-{kappa}B signaling in mediating tau toxicity in tauopathy.
]]></description>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Fan, L.</dc:creator>
<dc:creator>Zhan, L.</dc:creator>
<dc:creator>Kodama, L.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:creator>Chin, M. Y.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Le, D.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Mok, S.-A.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:date>2021-02-22</dc:date>
<dc:identifier>doi:10.1101/2021.02.22.432272</dc:identifier>
<dc:title><![CDATA[Microglial NF-κB drives tau spreading and toxicity in a mouse model of tauopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.25.432762v1?rss=1">
<title>
<![CDATA[
The type 2 asthma mediator IL-13 inhibits SARS-CoV-2 infection of bronchial epithelium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.25.432762v1?rss=1</link>
<description><![CDATA[
RationaleAsthma is associated with chronic changes in the airway epithelium, a key target of SARS-CoV-2. Many epithelial changes are driven by the type 2 cytokine IL-13, but the effects of IL-13 on SARS-CoV-2 infection are unknown.

ObjectivesWe sought to discover how IL-13 and other cytokines affect expression of genes encoding SARS-CoV-2-associated host proteins in human bronchial epithelial cells (HBECs) and determine whether IL-13 stimulation alters susceptibility to SARS-CoV-2 infection.

MethodsWe used bulk and single cell RNA-seq to identify cytokine-induced changes in SARS-CoV-2-associated gene expression in HBECs. We related these to gene expression changes in airway epithelium from individuals with mild-moderate asthma and chronic obstructive pulmonary disease (COPD). We analyzed effects of IL-13 on SARS-CoV-2 infection of HBECs.

Measurements and Main ResultsTranscripts encoding 332 of 342 (97%) SARS-CoV-2-associated proteins were detected in HBECs ([&ge;]1 RPM in 50% samples). 41 (12%) of these mRNAs were regulated by IL-13 (>1.5-fold change, FDR < 0.05). Many IL-13-regulated SARS-CoV-2-associated genes were also altered in type 2 high asthma and COPD. IL-13 pretreatment reduced viral RNA recovered from SARS-CoV-2 infected cells and decreased dsRNA, a marker of viral replication, to below the limit of detection in our assay. Mucus also inhibited viral infection.

ConclusionsIL-13 markedly reduces susceptibility of HBECs to SARS-CoV-2 infection through mechanisms that likely differ from those activated by type I interferons. Our findings may help explain reports of relatively low prevalence of asthma in patients diagnosed with COVID-19 and could lead to new strategies for reducing SARS-CoV-2 infection.
]]></description>
<dc:creator>Bonser, L. R.</dc:creator>
<dc:creator>Eckalbar, W. L.</dc:creator>
<dc:creator>Rodriguez, L.</dc:creator>
<dc:creator>Shen, J.</dc:creator>
<dc:creator>Koh, K. D.</dc:creator>
<dc:creator>Zlock, L. T.</dc:creator>
<dc:creator>Christenson, S.</dc:creator>
<dc:creator>Woodruff, P. G.</dc:creator>
<dc:creator>Finkbeiner, W. E.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:date>2021-02-25</dc:date>
<dc:identifier>doi:10.1101/2021.02.25.432762</dc:identifier>
<dc:title><![CDATA[The type 2 asthma mediator IL-13 inhibits SARS-CoV-2 infection of bronchial epithelium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.26.433123v1?rss=1">
<title>
<![CDATA[
A unicellular walker embodies a finite state machine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.26.433123v1?rss=1</link>
<description><![CDATA[
Cells are complex biochemical systems whose behavior emerges from interactions among myriad molecular components. Computation is often invoked as a general framework for navigating this cellular complexity. However, it is unclear how cells might embody computational processes such that theories of computation, including finite state machine models, could be productively applied. Here, we demonstrate finite state machine-like processing embodied in cells using the walking behavior of Euplotes eurystomus, a ciliate that walks across surfaces using fourteen motile appendages (cirri). We found that cellular walking entails regulated transitions between a discrete set of gait states. The set of observed transitions decomposes into a small group of high-probability, temporally irreversible transitions and a large group of low-probability time-symmetric transitions, thus revealing stereotypy in sequential patterns of state transitions. Simulations and experiments suggest that the sequential logic of the gait is functionally important. Taken together, these findings implicate a finite state machine-like process. Cirri are connected by microtubule bundles (fibers), and we found that the dynamics of cirri involved in different state transitions are associated with the structure of the fiber system. Perturbative experiments revealed that the fibers mediate gait coordination, suggesting a mechanical basis of gait control.
]]></description>
<dc:creator>Larson, B. T.</dc:creator>
<dc:creator>Garbus, J.</dc:creator>
<dc:creator>Pollack, J. B.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2021-02-27</dc:date>
<dc:identifier>doi:10.1101/2021.02.26.433123</dc:identifier>
<dc:title><![CDATA[A unicellular walker embodies a finite state machine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.28.433290v1?rss=1">
<title>
<![CDATA[
Infusion of CCR5 Gene-Edited T Cells Allows Immune Reconstitution, HIV Reservoir Decay, and Long-Term Virological Control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.28.433290v1?rss=1</link>
<description><![CDATA[
Antiretroviral therapy (ART) fails to fully restore immune function and is not curative. A single infusion of CCR5 gene-edited autologous CD4+ T cells (SB-728-T) led to sustained increases in CD4+ T cell counts, improved T cell homeostasis, and reduced the estimated size of the HIV reservoir. These outcomes were associated with the expansion and long-term persistence of a novel CCR5 gene-edited CD4+ T memory stem cell (CD45RAintROint TSCM) subset that can replenish the pool of more differentiated memory cells. We showed that novel CD45RAintROint TSCM cells are transcriptionally distinct from the previously described CD45RA+ TSCM and are minimally differentiated cells uncommitted to a specific Th-lineage. Subsequently, we showed in an independent trial that infusion of the SB-728-T cell product resulted in partial control of viral replication upon cessation of ART which was correlated with the frequencies of CCR5 gene-edited TSCM and their TEM progeny. Interestingly, one participant that remained off ART to this date demonstrated long-term maintenance of CCR5 gene-edited cells and increased frequency of polyfunctional HIV-specific CD4+ and CD8+ T cells, contributing to low levels of viral load 5 years post-infusion. Consequently, the generation of HIV protected memory CD4+ T cells by CCR5 disruption can contribute toward novel interventions aimed at achieving a sustained ART-free viral remission of HIV disease.
]]></description>
<dc:creator>Zeidan, J.</dc:creator>
<dc:creator>Sharma, A. A.</dc:creator>
<dc:creator>Lee, G.</dc:creator>
<dc:creator>Raad, A.</dc:creator>
<dc:creator>Fromentin, R.</dc:creator>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Ghneim, K.</dc:creator>
<dc:creator>Sanchez, G. P.</dc:creator>
<dc:creator>Benne, C.</dc:creator>
<dc:creator>Canderan, G.</dc:creator>
<dc:creator>Procopio, F. A.</dc:creator>
<dc:creator>Balderas, R.</dc:creator>
<dc:creator>Monette, G.</dc:creator>
<dc:creator>Lalezari, J. P.</dc:creator>
<dc:creator>Heffernan, J. M.</dc:creator>
<dc:creator>Sabbagh, L.</dc:creator>
<dc:creator>Chomont, N.</dc:creator>
<dc:creator>Ando, D.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Sekaly, R.-P.</dc:creator>
<dc:date>2021-03-01</dc:date>
<dc:identifier>doi:10.1101/2021.02.28.433290</dc:identifier>
<dc:title><![CDATA[Infusion of CCR5 Gene-Edited T Cells Allows Immune Reconstitution, HIV Reservoir Decay, and Long-Term Virological Control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.02.433644v1?rss=1">
<title>
<![CDATA[
Ribosomal RNA methylation by GidB is a capacitor for discrimination of mischarged tRNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.02.433644v1?rss=1</link>
<description><![CDATA[
Despite redundant cellular pathways to minimize translational errors, errors in protein synthesis are common. Pathways and mechanisms to minimize errors are classified as pre-ribosomal or ribosomal. Pre-ribosomal pathways are primarily concerned with the appropriate charging of tRNAs with their cognate amino acid. By contrast, the ribosomal decoding centre is considered  blind to mischarged tRNAs since these have cognate codon*anti-codon pairing. Here, we identified that in mycobacteria, deletion of the 16S ribosomal RNA methyltransferase gidB led to increased ribosomal discrimination of mischarged tRNAs. Discrimination only occurred in mycobacteria enriched from environments or genetic backgrounds with high rates of mistranslation. GidB deletion was necessary but not sufficient for reducing mistranslation due to misacylation. Analysis of new cryoEM structures of the M. smegmatis ribosomes derived from wild-type and gidB-deleted strains point to the interaction between the base methylated by GidB on the 16S RNA and an asparagine on the ribosomal S12 protein that when mistranslated to aspartate may be involved in altering translational fidelity. Our data suggest a mechanism by which mycobacterial ribosomes can discriminate mischarged tRNAs and that 16S rRNA differential methylation by GidB may act to prevent catastrophic translational error.
]]></description>
<dc:creator>Bi, Z.</dc:creator>
<dc:creator>Su, H.</dc:creator>
<dc:creator>Hong, J.</dc:creator>
<dc:creator>Javid, B.</dc:creator>
<dc:date>2021-03-03</dc:date>
<dc:identifier>doi:10.1101/2021.03.02.433644</dc:identifier>
<dc:title><![CDATA[Ribosomal RNA methylation by GidB is a capacitor for discrimination of mischarged tRNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.02.433652v1?rss=1">
<title>
<![CDATA[
Opposing motors provide mechanical and functional robustness in the human spindle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.02.433652v1?rss=1</link>
<description><![CDATA[
At each cell division, the spindle self-organizes from microtubules and motors. How the spindles diverse motors, often acting redundantly or in opposition, collectively give rise to its emergent architecture, mechanics, and function is unknown. In human spindles, the motors dynein and Eg5 generate contractile and extensile stress, respectively. Inhibiting dynein or its targeting factor NuMA leads to unfocused, turbulent spindles and inhibiting Eg5 leads to monopoles, yet bipolar spindles form when both are inhibited together. What, then, are the roles of these opposing motors? Here we generate NuMA/dynein- and Eg5-doubly inhibited spindles that not only attain a typical metaphase shape and size, but also undergo anaphase. However, these spindles have reduced microtubule dynamics and are mechanically fragile, fracturing under force. Further, they exhibit lagging chromosomes and dramatic left-handed twist at anaphase. Thus, while these opposing motor activities are not required for the spindles shape, they are essential to its mechanical and functional robustness. Together, this work suggests a design principle whereby opposing active stresses provide robustness to force-generating cellular structures.
]]></description>
<dc:creator>Neahring, L.</dc:creator>
<dc:creator>Cho, N. H.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2021-03-02</dc:date>
<dc:identifier>doi:10.1101/2021.03.02.433652</dc:identifier>
<dc:title><![CDATA[Opposing motors provide mechanical and functional robustness in the human spindle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.03.433701v1?rss=1">
<title>
<![CDATA[
CaSR modulates sodium channel-mediated Ca 2+ -dependent excitability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.03.433701v1?rss=1</link>
<description><![CDATA[
Increasing extracellular [Ca2+] ([Ca2+]o) strongly decreases intrinsic excitability in neurons but the mechanism is unclear. By one hypothesis, [Ca2+]o screens surface charge reducing voltage-dependent sodium channel (VGSC) activation and by another [Ca2+]o activates Calcium-sensing receptor (CaSR) closing the sodium-leak channel (NALCN). Here we report that action potential (AP) firing rates increased in wild-type (WT), but not CaSR null mutant (Casr-/-) neocortical neurons, following the switch from physiological to reduced Ca2+-containing Tyrode. However, after membrane potential correction, AP firing increased similarly in both genotypes inconsistent with CaSR regulation of NALCN. Activation of VGSCs was the dominant contributor to the increase in excitability after the [Ca2+]o change. VGSC conductance-voltage relationships were hyperpolarized by decreasing [Ca2+]o for Casr-/- neurons indicating CaSR contributes to [Ca2+]o-dependent excitability via VGSCs. Regulation of VGSC gating by [Ca2+]o is the key mechanism mediating [Ca2+]o-dependent changes in neocortical neuron excitability and CaSR influences neuronal excitability by its effects on VGSC gating.
]]></description>
<dc:creator>Martiszus, B. J.</dc:creator>
<dc:creator>Tsintsadze, T.</dc:creator>
<dc:creator>Smith, S. M.</dc:creator>
<dc:creator>Chang, W.</dc:creator>
<dc:date>2021-03-04</dc:date>
<dc:identifier>doi:10.1101/2021.03.03.433701</dc:identifier>
<dc:title><![CDATA[CaSR modulates sodium channel-mediated Ca 2+ -dependent excitability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.04.433985v1?rss=1">
<title>
<![CDATA[
Large-scale placenta DNA methylation mega-analysis reveals fetal sex-specific differentially methylated CpG sites and regions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.04.433985v1?rss=1</link>
<description><![CDATA[
Although male-female differences in placental structure and function have been observed, little is understood about their molecular underpinnings. Here, we present a mega-analysis of 14 publicly available placenta DNA methylation (DNAm) microarray datasets to identify individual CpGs and regions associated with fetal sex. In the discovery dataset of placentas from full term pregnancies (N = 532 samples), 5,212 CpGs met genome-wide significance (p < 1E-8) and were enriched in pathways such as keratinization (FDR p-value = 7.37E-14), chemokine activity (FDR p-value = 1.56E-2), and eosinophil migration (FDR p-value = 1.83E-2). Nine differentially methylated regions were identified (fwerArea < 0.1) including a region in the promoter of ZNF300 that showed consistent differential DNAm in samples from earlier timepoints in pregnancy and appeared to be driven predominately by effects in the trophoblast cell type. We describe the largest study of fetal sex differences in placenta DNAm performed to date, revealing genes and pathways characterizing sex-specific placenta function and health outcomes later in life.
]]></description>
<dc:creator>Andrews, S. V.</dc:creator>
<dc:creator>Yang, I. J.</dc:creator>
<dc:creator>Froehlich, K.</dc:creator>
<dc:creator>Oskotsky, T.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2021-03-05</dc:date>
<dc:identifier>doi:10.1101/2021.03.04.433985</dc:identifier>
<dc:title><![CDATA[Large-scale placenta DNA methylation mega-analysis reveals fetal sex-specific differentially methylated CpG sites and regions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.06.434199v1?rss=1">
<title>
<![CDATA[
Elevated N-glycosylation of immunoglobulin G variable regions in myasthenia gravis highlights a commonality across autoantibody-associated diseases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.06.434199v1?rss=1</link>
<description><![CDATA[
Elevated N-linked glycosylation of immunoglobulin G variable regions (IgG-VN-Glyc) is an emerging molecular phenotype associated with autoimmune disorders. To test the broader specificity of elevated IgG-VN-Glyc, we studied patients with distinct subtypes of myasthenia gravis (MG), a B cell-mediated autoimmune disease. Our experimental design included adaptive immune receptor repertoire sequencing to quantify and characterize N-glycosylation sites in the global B cell receptor repertoire, proteomics to examine glycosylation patterns of the circulating IgG, and production of human-derived recombinant autoantibodies, which were studied with mass spectrometry and antigen binding assays to confirm occupation of glycosylation sites and determine whether they alter binding. We found that the frequency of IgG-VN-Glyc motifs was increased in the B cell repertoire of MG patients when compared to healthy donors. Motifs were introduced by both biased V gene segment usage and somatic hypermutation. IgG-VN-Glyc could be observed in the circulating IgG in a subset of MG patients. Autoantigen binding, by patient-derived MG autoantigen-specific monoclonal antibodies with experimentally confirmed presence of IgG-VN-Glyc, was not altered by the glycosylation. Our findings extend prior work on patterns of variable region N-linked glycosylation in autoimmunity to MG subtypes. Although occupied IgG-VN-Glyc motifs are found on MG autoantigen-specific monoclonal antibodies, they are not required for binding to the autoantigen in this disease.
]]></description>
<dc:creator>Mandel-Brehm, C.</dc:creator>
<dc:creator>Fichtner, M. L.</dc:creator>
<dc:creator>Jiang, R.</dc:creator>
<dc:creator>Winton, V. J.</dc:creator>
<dc:creator>Vazquez, S. E.</dc:creator>
<dc:creator>Pham, M. C.</dc:creator>
<dc:creator>Hoehn, K. B.</dc:creator>
<dc:creator>Kelleher, N. L.</dc:creator>
<dc:creator>Nowak, R. J.</dc:creator>
<dc:creator>Kleinstein, S. H.</dc:creator>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>OConnor, K. C.</dc:creator>
<dc:date>2021-03-08</dc:date>
<dc:identifier>doi:10.1101/2021.03.06.434199</dc:identifier>
<dc:title><![CDATA[Elevated N-glycosylation of immunoglobulin G variable regions in myasthenia gravis highlights a commonality across autoantibody-associated diseases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.08.434485v1?rss=1">
<title>
<![CDATA[
Stromal inflammation is a targetable driver of hematopoietic aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.08.434485v1?rss=1</link>
<description><![CDATA[
Hematopoietic aging is marked by a loss of regenerative capacity and skewed differentiation from hematopoietic stem cells (HSC) leading to dysfunctional blood production. Signals from the bone marrow (BM) niche dynamically tailor hematopoiesis, but the effect of aging on the niche microenvironment and the contribution of the aging niche to blood aging still remains unclear. Here, we characterize the inflammatory milieu in the aged marrow cavity that drives both stromal and hematopoietic remodeling. We find decreased numbers and functionality of osteogenic mesenchymal stromal cells (MSC) at the endosteum and expansion of pro-inflammatory perisinusoidal MSCs with deterioration of sinusoidal endothelium in the central marrow, which together create a degraded and inflamed old niche. Molecular mapping at single cell resolution confirms disruption of cell identities and enrichment of inflammatory response genes in niche populations. Niche inflammation, in turn, drives chronic activation of emergency myelopoiesis pathways in old HSCs and multipotent progenitors (MPP), which promotes myeloid differentiation at the expense of lymphoid and erythroid commitment and hinders hematopoietic regeneration. Remarkably, niche deterioration, HSC dysfunction and defective hematopoietic regeneration, can be improved by blocking inflammatory IL-1 signaling. Our results demonstrate that targeting niche inflammation is a tractable strategy to restore blood production during aging.
]]></description>
<dc:creator>Verovskaya, E. V.</dc:creator>
<dc:creator>Mitchell, C. A.</dc:creator>
<dc:creator>Calero-Nieto, F. J.</dc:creator>
<dc:creator>Herault, A.</dc:creator>
<dc:creator>Dellorusso, P. D.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Zhang, S. Y.</dc:creator>
<dc:creator>Flohr Svendsen, A.</dc:creator>
<dc:creator>Pietras, E. M.</dc:creator>
<dc:creator>Bakker, S. T.</dc:creator>
<dc:creator>Ho, T. T.</dc:creator>
<dc:creator>Gottgens, B.</dc:creator>
<dc:creator>Passegue, E.</dc:creator>
<dc:date>2021-03-09</dc:date>
<dc:identifier>doi:10.1101/2021.03.08.434485</dc:identifier>
<dc:title><![CDATA[Stromal inflammation is a targetable driver of hematopoietic aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.09.434529v1?rss=1">
<title>
<![CDATA[
Longitudinal single-cell epitope and RNA-sequencing reveals the immunological impact of type 1 interferon autoantibodies in critical COVID-19 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.09.434529v1?rss=1</link>
<description><![CDATA[
Type I interferon (IFN-I) neutralizing autoantibodies have been found in some critical COVID-19 patients; however, their prevalence and longitudinal dynamics across the disease severity scale, and functional effects on circulating leukocytes remain unknown. Here, in 284 COVID-19 patients, we found IFN-I autoantibodies in 19% of critical, 6% of severe and none of the moderate cases. Longitudinal profiling of over 600,000 peripheral blood mononuclear cells using multiplexed single-cell epitope and transcriptome sequencing from 54 COVID-19 patients, 15 non-COVID-19 patients and 11 non-hospitalized healthy controls, revealed a lack of IFN-I stimulated gene (ISG-I) response in myeloid cells from critical cases, including those producing anti-IFN-I autoantibodies. Moreover, surface protein analysis showed an inverse correlation of the inhibitory receptor LAIR-1 with ISG-I expression response early in the disease course. This aberrant ISG-I response in critical patients with and without IFN-I autoantibodies, supports a unifying model for disease pathogenesis involving ISG-I suppression via convergent mechanisms.
]]></description>
<dc:creator>van der Wijst, M. G. P.</dc:creator>
<dc:creator>Vazquez, S. E.</dc:creator>
<dc:creator>Hartoularos, G. C.</dc:creator>
<dc:creator>Bastard, P.</dc:creator>
<dc:creator>Grant, T.</dc:creator>
<dc:creator>Bueno, R.</dc:creator>
<dc:creator>Lee, D. S.</dc:creator>
<dc:creator>Greenland, J. R.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Perez, R.</dc:creator>
<dc:creator>Ogorodnikov, A.</dc:creator>
<dc:creator>Ward, A.</dc:creator>
<dc:creator>Mann, S. A.</dc:creator>
<dc:creator>Lynch, K. L.</dc:creator>
<dc:creator>Yun, C.</dc:creator>
<dc:creator>Havlir, D. V.</dc:creator>
<dc:creator>Chamie, G.</dc:creator>
<dc:creator>Marquez, C.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:creator>Lionakis, M. S.</dc:creator>
<dc:creator>Norris, P. J.</dc:creator>
<dc:creator>Dumont, L. J.</dc:creator>
<dc:creator>Kelly, K.</dc:creator>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Gervais, A.</dc:creator>
<dc:creator>Le Voyer, T.</dc:creator>
<dc:creator>Whatley, A.</dc:creator>
<dc:creator>Si, Y.</dc:creator>
<dc:creator>Byrne, A.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Arkal, A.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>UCSF COMET consortium,</dc:creator>
<dc:creator>Kangelaris, K.</dc:creator>
<dc:creator>Calfee, C. S.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:creator>Hendrickson, C.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Woodruff, P. G.</dc:creator>
<dc:creator>Langelier, C. R.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2021-03-10</dc:date>
<dc:identifier>doi:10.1101/2021.03.09.434529</dc:identifier>
<dc:title><![CDATA[Longitudinal single-cell epitope and RNA-sequencing reveals the immunological impact of type 1 interferon autoantibodies in critical COVID-19]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.12.435197v1?rss=1">
<title>
<![CDATA[
Massively parallel reporter perturbation assay uncovers temporal regulatory architecture during neural differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.12.435197v1?rss=1</link>
<description><![CDATA[
Gene regulatory elements play a key role in orchestrating gene expression during cellular differentiation, but what determines their function over time remains largely unknown. Here, we performed perturbation-based massively parallel reporter assays at seven early time points of neural differentiation to systematically characterize how regulatory elements and motifs within them guide cellular differentiation. By perturbing over 2,000 putative DNA binding motifs in active regulatory regions, we delineated four categories of functional elements, and observed that activity direction is mostly determined by the sequence itself, while the magnitude of effect depends on the cellular environment. We also find that fine-tuning transcription rates is often achieved by a combined activity of adjacent activating and repressing elements. Our work provides a blueprint for the sequence components needed to induce different transcriptional patterns in general and specifically during neural differentiation.
]]></description>
<dc:creator>Kreimer, A.</dc:creator>
<dc:creator>Ashuach, T.</dc:creator>
<dc:creator>Inoue, F.</dc:creator>
<dc:creator>Khodaverdian, A.</dc:creator>
<dc:creator>Yosef, N.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2021-03-12</dc:date>
<dc:identifier>doi:10.1101/2021.03.12.435197</dc:identifier>
<dc:title><![CDATA[Massively parallel reporter perturbation assay uncovers temporal regulatory architecture during neural differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.14.435332v1?rss=1">
<title>
<![CDATA[
Community Evaluation of Glycoproteomics Informatics Solutions Reveals High-Performance Search Strategies of Glycopeptide Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.14.435332v1?rss=1</link>
<description><![CDATA[
Glycoproteome profiling (glycoproteomics) is a powerful yet analytically challenging research tool. The complex tandem mass spectra generated from glycopeptide mixtures require sophisticated analysis pipelines for structural determination. Diverse software aiding the process have appeared, but their relative performance remains untested. Conducted through the HUPO Human Proteome Project - Human Glycoproteomics Initiative, this community study, comprising both developers and users of glycoproteomics software, evaluates the performance of informatics solutions for system-wide glycopeptide analysis. Mass spectrometry-based glycoproteomics datasets from human serum were shared with all teams. The relative team performance for N- and O-glycopeptide data analysis was comprehensively established and validated through orthogonal performance tests. Excitingly, several high-performance glycoproteomics informatics solutions were identified. While the study illustrated that significant informatics challenges remain, as indicated by a high discordance between annotated glycopeptides, lists of high-confidence (consensus) glycopeptides were compiled from the standardised team reports. Deep analysis of the performance data revealed key performance-associated search variables and led to recommendations for improved "high coverage" and "high accuracy" glycoproteomics search strategies. This study concludes that diverse software for comprehensive glycopeptide data analysis exist, points to several high-performance search strategies, and specifies key variables that may guide future software developments and assist informatics decision-making in glycoproteomics.
]]></description>
<dc:creator>Kawahara, R.</dc:creator>
<dc:creator>Alagesan, K.</dc:creator>
<dc:creator>Bern, M.</dc:creator>
<dc:creator>Cao, W.</dc:creator>
<dc:creator>Chalkley, R. J.</dc:creator>
<dc:creator>Cheng, K.</dc:creator>
<dc:creator>Choo, M. S.</dc:creator>
<dc:creator>Edwards, N.</dc:creator>
<dc:creator>Goldman, R.</dc:creator>
<dc:creator>Hoffmann, M.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Kim, J. Y.</dc:creator>
<dc:creator>Kletter, D.</dc:creator>
<dc:creator>Liquet-Weiland, B.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Mechref, Y.</dc:creator>
<dc:creator>Meng, B.</dc:creator>
<dc:creator>Neelamegham, S.</dc:creator>
<dc:creator>Nguyen-Khuong, T.</dc:creator>
<dc:creator>Nilsson, J.</dc:creator>
<dc:creator>Pap, A.</dc:creator>
<dc:creator>Park, G. W.</dc:creator>
<dc:creator>Parker, B. L.</dc:creator>
<dc:creator>Pegg, C. L.</dc:creator>
<dc:creator>Penninger, J. M.</dc:creator>
<dc:creator>Phung, T. K.</dc:creator>
<dc:creator>Pioch, M.</dc:creator>
<dc:creator>Rapp, E.</dc:creator>
<dc:creator>Sakalli, E.</dc:creator>
<dc:creator>Sanda, M.</dc:creator>
<dc:creator>Shulz, B. L.</dc:creator>
<dc:creator>Scott, N. E.</dc:creator>
<dc:creator>Sofronov, G.</dc:creator>
<dc:creator>Stadlmann, J.</dc:creator>
<dc:creator>Vakhrushev, S. Y.</dc:creator>
<dc:creator>Woo, C. M.</dc:creator>
<dc:creator>Wu, H.-Y.</dc:creator>
<dc:creator>Yang, P.</dc:creator>
<dc:creator>Ying, W.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Zaia, J.</dc:creator>
<dc:creator>Haslam, S. M.</dc:creator>
<dc:creator>Palmisano, G.</dc:creator>
<dc:creator>Yoo, J. S</dc:creator>
<dc:date>2021-03-15</dc:date>
<dc:identifier>doi:10.1101/2021.03.14.435332</dc:identifier>
<dc:title><![CDATA[Community Evaluation of Glycoproteomics Informatics Solutions Reveals High-Performance Search Strategies of Glycopeptide Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.16.435520v1?rss=1">
<title>
<![CDATA[
Noncanonical mRNA decay by the endoplasmic-reticulum stress sensor IRE1α promotes cancer-cell survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.16.435520v1?rss=1</link>
<description><![CDATA[
Eukaryotic IRE1 mitigates endoplasmic-reticulum (ER) stress by orchestrating the unfolded-protein response (UPR). IRE1 spans the ER membrane, and signals through a cytosolic kinase-endoribonuclease module. The endoribonuclease generates the transcription factor XBP1s by intron excision between similar RNA stem-loop endomotifs, and depletes select cellular mRNAs through regulated IRE1-dependent decay (RIDD). Paradoxically, mammalian RIDD seemingly targets only mRNAs with XBP1-like endomotifs, while in flies RIDD exhibits little sequence restriction. By comparing nascent and total IRE1-controlled mRNAs in human breast cancer cells, we discovered not only canonical endomotif-containing RIDD substrates, but also many targets lacking recognizable motifs--degraded by a process we coin RIDDLE, for RIDD lacking endomotif. IRE1 displayed two basic endoribonuclease modalities: endomotif-specific cleavage, minimally requiring dimers; and endomotif-independent promiscuous processing, requiring phospho-oligomers. An oligomer-deficient mutant that did not support RIDDLE failed to rescue cancer-cell viability. These results link IRE1 oligomers, RIDDLE, and cell survival, advancing mechanistic understanding of the UPR.
]]></description>
<dc:creator>Le Thomas, A.</dc:creator>
<dc:creator>Ferri, E.</dc:creator>
<dc:creator>Marsters, S.</dc:creator>
<dc:creator>Harnoss, J. M.</dc:creator>
<dc:creator>Modrusan, Z.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Rudolph, J.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Wu, T. D.</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:creator>Ashkenazi, A.</dc:creator>
<dc:date>2021-03-16</dc:date>
<dc:identifier>doi:10.1101/2021.03.16.435520</dc:identifier>
<dc:title><![CDATA[Noncanonical mRNA decay by the endoplasmic-reticulum stress sensor IRE1α promotes cancer-cell survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.18.436011v1?rss=1">
<title>
<![CDATA[
Tight nanoscale clustering of Fc gamma-receptors using DNA origami promotes phagocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.18.436011v1?rss=1</link>
<description><![CDATA[
Macrophages destroy pathogens and diseased cells through Fc{gamma} receptor (Fc{gamma}R)-driven phagocytosis of antibody-opsonized targets. Phagocytosis requires activation of multiple Fc{gamma}Rs, but the mechanism controlling the threshold for response is unclear. We developed a DNA origami-based engulfment system that allows precise nanoscale control of the number and spacing of ligands. When the number of ligands remains constant, reducing ligand spacing from 17.5 nm to 7 nm potently enhances engulfment, primarily by increasing efficiency of the engulfment-initiation process. Tighter ligand clustering increases receptor phosphorylation, as well as proximal downstream signals. Increasing the number of signaling domains recruited to a single ligand-receptor complex was not sufficient to recapitulate this effect, indicating that clustering of multiple receptors is required. Our results suggest that macrophages use information about local ligand densities to make critical engulfment decisions, which has implications for the mechanism of antibody-mediated phagocytosis and the design of immunotherapies.
]]></description>
<dc:creator>Kern, N.</dc:creator>
<dc:creator>Dong, R.</dc:creator>
<dc:creator>Douglas, S. M.</dc:creator>
<dc:creator>Vale, R. D.</dc:creator>
<dc:creator>Morrissey, M. A.</dc:creator>
<dc:date>2021-03-18</dc:date>
<dc:identifier>doi:10.1101/2021.03.18.436011</dc:identifier>
<dc:title><![CDATA[Tight nanoscale clustering of Fc gamma-receptors using DNA origami promotes phagocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.18.436057v1?rss=1">
<title>
<![CDATA[
Ciliary Hedgehog signaling regulates cell survival to build the facial midline 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.18.436057v1?rss=1</link>
<description><![CDATA[
Craniofacial defects are among the most common phenotypes caused by ciliopathies, yet the developmental and molecular etiology of these defects is poorly understood. We investigated multiple mouse models of human ciliopathies (including Tctn2, Cc2d2a and Tmem231 mutants) and discovered that each displays hypotelorism, a narrowing of the midface. As early in development as the end of gastrulation, Tctn2 mutants displayed reduced activation of the Hedgehog (HH) pathway in the prechordal plate, the head organizer. This prechordal plate defect preceded a reduction of HH pathway activation and Shh expression in the adjacent neurectoderm. Concomitant with the reduction of HH pathway activity, Tctn2 mutants exhibited increased cell death in the neurectoderm and facial ectoderm, culminating in a collapse of the facial midline. Enhancing HH signaling by decreasing the gene dosage of a negative regulator of the pathway, Ptch1, decreased cell death and rescued the midface defect in both Tctn2 and Cc2d2a mutants. These results reveal that ciliary HH signaling mediates communication between the prechordal plate and the neurectoderm to provide cellular survival cues essential for development of the facial midline.
]]></description>
<dc:creator>Abrams, S. R.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:date>2021-03-18</dc:date>
<dc:identifier>doi:10.1101/2021.03.18.436057</dc:identifier>
<dc:title><![CDATA[Ciliary Hedgehog signaling regulates cell survival to build the facial midline]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.19.436138v1?rss=1">
<title>
<![CDATA[
Intraflagellar transport trains can turn around without the ciliary tip complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.19.436138v1?rss=1</link>
<description><![CDATA[
Cilia and flagella are microtubule doublet based organelles found across the eukaryotic tree of life. Their very high aspect ratio and crowded interior are unfavourable to diffusive transport for their assembly and maintenance. Instead, a highly dynamic system of intraflagellar transport (IFT) trains moves rapidly up and down the cilium. However, the mechanism of how these trains turn around upon reaching the ciliary tip has remained elusive. It has been hypothesized that there exists a dedicated calcium-dependent protein-based machinery at the ciliary tip to mediate this conversion. In this work, we use physical and chemical methods to manipulate IFT in the cilia of the unicellular green alga Chlamydomonas reinhardtii to show that no such stationary tip-machinery is required for IFT turnaround. Instead, we demonstrate that the conversion from anterograde to retrograde IFT trains is a calcium independent intrinsic ability of the IFT system.
]]></description>
<dc:creator>Nievergelt, A. P.</dc:creator>
<dc:creator>Zykov, I.</dc:creator>
<dc:creator>Diener, D. R.</dc:creator>
<dc:creator>Buchholz, T.-O.</dc:creator>
<dc:creator>Delling, M.</dc:creator>
<dc:creator>Diez, S.</dc:creator>
<dc:creator>Jug, F.</dc:creator>
<dc:creator>Stepanek, L.</dc:creator>
<dc:creator>Pigino, G.</dc:creator>
<dc:date>2021-03-19</dc:date>
<dc:identifier>doi:10.1101/2021.03.19.436138</dc:identifier>
<dc:title><![CDATA[Intraflagellar transport trains can turn around without the ciliary tip complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.19.436190v1?rss=1">
<title>
<![CDATA[
Coupling of spliceosome complexity to intron diversity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.19.436190v1?rss=1</link>
<description><![CDATA[
We determined that over 40 spliceosomal proteins are conserved between many fungal species and humans but were lost during the evolution of S. cerevisiae, an intron-poor yeast with unusually rigid splicing signals. We analyzed null mutations in a subset of these factors, most of which had not been investigated previously, in the intron-rich yeast Cryptococcus neoformans. We found they govern splicing efficiency of introns with divergent spacing between intron elements. Importantly, most of these factors also suppress usage of weak nearby cryptic/alternative splice sites. Among these, orthologs of GPATCH1 and the helicase DHX35 display correlated functional signatures and copurify with each other as well as components of catalytically active spliceosomes, identifying a conserved G-patch/helicase pair that promotes splicing fidelity. We propose that a significant fraction of spliceosomal proteins in humans and most eukaryotes are involved in limiting splicing errors, potentially through kinetic proofreading mechanisms, thereby enabling greater intron diversity.
]]></description>
<dc:creator>Sales-Lee, J.</dc:creator>
<dc:creator>Perry, D. S.</dc:creator>
<dc:creator>Bowser, B. A.</dc:creator>
<dc:creator>Diedrich, J. K.</dc:creator>
<dc:creator>Rao, B.</dc:creator>
<dc:creator>Beusch, I.</dc:creator>
<dc:creator>Yates, J. R.</dc:creator>
<dc:creator>Roy, S. W.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:date>2021-03-20</dc:date>
<dc:identifier>doi:10.1101/2021.03.19.436190</dc:identifier>
<dc:title><![CDATA[Coupling of spliceosome complexity to intron diversity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.19.436221v1?rss=1">
<title>
<![CDATA[
Anti-GD2 antibody disrupts GD2:Siglec-7 interactions and synergizes with CD47 blockade to mediate tumor eradication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.19.436221v1?rss=1</link>
<description><![CDATA[
The disialoganglioside GD2 is consistently overexpressed in neuroblastoma and osteosarcoma, and is variably expressed in other sarcomas, gliomas, neuroendocrine tumors, and epithelial cancers. Anti-GD2 antibodies have improved the survival rates of patients with neuroblastoma only when administered as part of intense chemotherapy-based cytotoxic regimens, which are associated with debilitating late effects including hearing loss, growth retardation, and secondary leukemias. Despite broad expression of GD2 on osteosarcoma, anti-GD2 antibody has not mediated significant antitumor activity in that disease or any other GD2+ cancers. CD47 is a checkpoint molecule overexpressed on tumor cells that inhibits macrophage activity, and CD47 blockade has demonstrated promising clinical activity in early human trials. We investigated whether anti-CD47 antibody could enhance the efficacy of anti-GD2 antibody in neuroblastoma and other GD2+ malignancies. We demonstrate substantial synergy of these two agents, resulting in the recruitment of tumor associated macrophages (TAMs) to mediate robust and durable anti-tumor responses. The responses are driven by GD2-specific factors that reorient the balance of macrophage activity towards phagocytosis of tumor cells, including disruption of a newly described GD2:Siglec-7 axis. These results demonstrate the unique synergy of combining anti-GD2 with anti-CD47, which has the potential to significantly enhance outcomes for children with neuroblastoma and osteosarcoma and will soon be investigated in a first-in-human clinical trial.
]]></description>
<dc:creator>Theruvath, J.</dc:creator>
<dc:creator>Menard, M.</dc:creator>
<dc:creator>Smith, B. A.</dc:creator>
<dc:creator>Linde, M. H.</dc:creator>
<dc:creator>Coles, G.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Kiru, L.</dc:creator>
<dc:creator>Delaidelli, A.</dc:creator>
<dc:creator>Silberstein, J. L.</dc:creator>
<dc:creator>Banuelos, A.</dc:creator>
<dc:creator>Dhingra, S.</dc:creator>
<dc:creator>Sotillo, E.</dc:creator>
<dc:creator>Heitzeneder, S.</dc:creator>
<dc:creator>Tousley, A.</dc:creator>
<dc:creator>Lattin, J.</dc:creator>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Huang, J.</dc:creator>
<dc:creator>Nasholm, N.</dc:creator>
<dc:creator>Dalton, G. N.</dc:creator>
<dc:creator>He, A.</dc:creator>
<dc:creator>Kuo, T. C.</dc:creator>
<dc:creator>Sangalang, E. R. B.</dc:creator>
<dc:creator>Pons, J.</dc:creator>
<dc:creator>Barkal, A.</dc:creator>
<dc:creator>Brewer, R.</dc:creator>
<dc:creator>Marjon, K. D.</dc:creator>
<dc:creator>Marshall, P. L.</dc:creator>
<dc:creator>Fernandes, R.</dc:creator>
<dc:creator>Cochran, J.</dc:creator>
<dc:creator>Sorensen, P.</dc:creator>
<dc:creator>Daldrup-Link, H. E.</dc:creator>
<dc:creator>Weissman, I. L.</dc:creator>
<dc:creator>Sage, J.</dc:creator>
<dc:creator>Majeti, R.</dc:creator>
<dc:creator>Bertozzi, C. R.</dc:creator>
<dc:creator>Weiss, W. A.</dc:creator>
<dc:creator>Mackall, C. L.</dc:creator>
<dc:creator>Majzner, R. G.</dc:creator>
<dc:date>2021-03-21</dc:date>
<dc:identifier>doi:10.1101/2021.03.19.436221</dc:identifier>
<dc:title><![CDATA[Anti-GD2 antibody disrupts GD2:Siglec-7 interactions and synergizes with CD47 blockade to mediate tumor eradication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.22.436511v1?rss=1">
<title>
<![CDATA[
Proteome plasticity in response to persistent environmental change 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.22.436511v1?rss=1</link>
<description><![CDATA[
Temperature is a variable component of the environment and all organisms must deal with or adapt to temperature change. Acute temperature change activates cellular stress responses resulting in the refolding or removal of damaged proteins. However, how organisms adapt to long-term temperature change remains largely unexplored. Here, we report that budding yeast responds to long-term high temperature challenge by switching from chaperone induction to the reduction of temperature sensitive proteins and re-localizing a portion of its proteome. Surprisingly, we also find many proteins adopt an alternative conformation. Using Fet3p as an example, we find that the temperature-dependent conformational difference is accompanied by distinct thermostability, subcellular localization, and importantly, cellular functions. We postulate that in addition to the known mechanisms of adaptation, conformational plasticity allows some polypeptides to acquire new biophysical properties and functions when environmental change endures.
]]></description>
<dc:creator>Domnauer, M.</dc:creator>
<dc:creator>Zheng, F.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Chang, C. E.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Conkright-Fincham, J.</dc:creator>
<dc:creator>McCroskey, S.</dc:creator>
<dc:creator>Florens, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Fong, B.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Ramanathan, A.</dc:creator>
<dc:creator>Si, K.</dc:creator>
<dc:creator>Zhou, C.</dc:creator>
<dc:date>2021-03-22</dc:date>
<dc:identifier>doi:10.1101/2021.03.22.436511</dc:identifier>
<dc:title><![CDATA[Proteome plasticity in response to persistent environmental change]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.23.436694v1?rss=1">
<title>
<![CDATA[
Condensin DC spreads linearly and bidirectionally from recruitment sites to create loop-anchored TADs in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.23.436694v1?rss=1</link>
<description><![CDATA[
Condensins are molecular motors that compact DNA via linear translocation. In C. elegans, the X-chromosome harbors a specialized condensin that participates in dosage compensation (DC). Condensin DC is recruited to and spreads from a small number of recruit elements on the X-chromosome (rex) and is required for the formation of topologically associating domains (TADs). We take advantage of autosomes that are largely devoid of condensin DC and TADs to address how rex sites and condensin DC give rise to the formation of TADs. When an autosome and X-chromosome are physically fused, despite the spreading of condensin DC into the autosome, no TAD was created. Insertion of a strong rex on the X-chromosome results in the TAD boundary formation regardless of sequence orientation. When the same rex is inserted on an autosome, despite condensin DC recruitment, there was no spreading or features of a TAD. On the other hand, when a "super rex" composed of six rex sites or three separate rex sites are inserted on an autosome, recruitment and spreading of condensin DC led to formation of TADs. Therefore, recruitment to and spreading from rex sites are necessary and sufficient for recapitulating loop-anchored TADs observed on the X-chromosome. Together our data suggest a model in which rex sites are both loading sites and bidirectional barriers for condensin DC, a one-sided loop-extruder with movable inactive anchor.
]]></description>
<dc:creator>Jimenez, D. S.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Ragipani, B.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Street, L. A.</dc:creator>
<dc:creator>Kramer, M.</dc:creator>
<dc:creator>Albritton, S. E.</dc:creator>
<dc:creator>Winterkorn, L.</dc:creator>
<dc:creator>Ercan, S.</dc:creator>
<dc:date>2021-03-24</dc:date>
<dc:identifier>doi:10.1101/2021.03.23.436694</dc:identifier>
<dc:title><![CDATA[Condensin DC spreads linearly and bidirectionally from recruitment sites to create loop-anchored TADs in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.01.438060v1?rss=1">
<title>
<![CDATA[
Time-varying Dynamic Network Model For Dynamic Resting State Functional Connectivity in fMRI and MEG imaging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.01.438060v1?rss=1</link>
<description><![CDATA[
Dynamic resting state functional connectivity (RSFC) characterizes fluctuations that occurs over time in functional brain networks. Existing methods to extract dynamic RSFCs, such as sliding-window and clustering methods, have various limitations due to their inherent non-adaptive nature and high-dimensionality including an inability to reconstruct brain signals, insufficiency of data for reliable estimation, insensitivity to rapid changes in dynamics, and a lack of generalizability across multimodal functional imaging datasets. To overcome these deficiencies, we develop a novel and unifying time-varying dynamic network (TVDN) framework for examining dynamic resting state functional connectivity. TVDN includes a generative model that describes the relation between low-dimensional dynamic RSFC and the brain signals, and an inference algorithm that automatically and adaptively learns to detect dynamic state transitions in data and a low-dimensional manifold of dynamic RSFC. TVDN is generalizable to handle multimodal functional neuroimaging data (fMRI and MEG/EEG). The resulting estimated low-dimensional dynamic RSFCs manifold directly links to the frequency content of brain signals. Hence we can evaluate TVDN performance by examining whether learnt features can reconstruct observed brain signals. We conduct comprehensive simulations to evaluate TVDN under hypothetical settings. We then demonstrate the application of TVDN with real fMRI and MEG data, and compare the results with existing benchmarks. Results demonstrate that TVDN is able to correctly capture the dynamics of brain activity and more robustly detect brain state switching both in resting state fMRI and MEG data.
]]></description>
<dc:creator>Jiang, F.</dc:creator>
<dc:creator>Jin, H.</dc:creator>
<dc:creator>Bao, Y.</dc:creator>
<dc:creator>Xie, X.</dc:creator>
<dc:creator>Cummings, J.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2021-04-02</dc:date>
<dc:identifier>doi:10.1101/2021.04.01.438060</dc:identifier>
<dc:title><![CDATA[Time-varying Dynamic Network Model For Dynamic Resting State Functional Connectivity in fMRI and MEG imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.02.438213v1?rss=1">
<title>
<![CDATA[
TDP-43 represses cryptic exon inclusion in FTD/ALS gene UNC13A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.02.438213v1?rss=1</link>
<description><![CDATA[
A hallmark pathological feature of neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) is the depletion of RNA-binding protein TDP-43 from the nucleus of neurons in the brain and spinal cord. A major function of TDP-43 is as a repressor of cryptic exon inclusion during RNA splicing. Single nucleotide polymorphisms (SNPs) in UNC13A are among the strongest genome-wide association study (GWAS) hits associated with FTD/ALS in humans, but how those variants increase risk for disease is unknown. Here we show that TDP-43 represses a cryptic exon splicing event in UNC13A. Loss of TDP-43 from the nucleus in human brain, neuronal cell lines, and iPSC-derived motor neurons resulted in the inclusion of a cryptic exon in UNC13A mRNA and reduced UNC13A protein expression. Remarkably, the top variants associated with FTD/ALS risk in humans are located in the cryptic exon harboring intron itself and we show that they increase UNC13A cryptic exon splicing in the face of TDP-43 dysfunction. Together, our data provide a direct functional link between one of the strongest genetic risk factors for FTD/ALS (UNC13A genetic variants) and loss of TDP-43 function.
]]></description>
<dc:creator>Ma, X. R.</dc:creator>
<dc:creator>Prudencio, M.</dc:creator>
<dc:creator>Koike, Y.</dc:creator>
<dc:creator>Vatsavayai, S. C.</dc:creator>
<dc:creator>Kim, G.</dc:creator>
<dc:creator>Harbinski, F.</dc:creator>
<dc:creator>Rodriguez, C. M.</dc:creator>
<dc:creator>Schmidt, H. B.</dc:creator>
<dc:creator>Cummings, B. B.</dc:creator>
<dc:creator>Wyatt, D. W.</dc:creator>
<dc:creator>Kurylo, K.</dc:creator>
<dc:creator>Miller, G.</dc:creator>
<dc:creator>Mekhoubad, S.</dc:creator>
<dc:creator>Sallee, N.</dc:creator>
<dc:creator>Jansen-West, K.</dc:creator>
<dc:creator>Cook, C.</dc:creator>
<dc:creator>Pickles, S.</dc:creator>
<dc:creator>Oskarsson, B.</dc:creator>
<dc:creator>Graff-Radford, N. R.</dc:creator>
<dc:creator>Boeve, B. F.</dc:creator>
<dc:creator>Knopman, D. S.</dc:creator>
<dc:creator>Petersen, R. C.</dc:creator>
<dc:creator>Dickson, D. W.</dc:creator>
<dc:creator>Green, E. M.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Petrucelli, L.</dc:creator>
<dc:creator>Gitler, A. D.</dc:creator>
<dc:date>2021-04-04</dc:date>
<dc:identifier>doi:10.1101/2021.04.02.438213</dc:identifier>
<dc:title><![CDATA[TDP-43 represses cryptic exon inclusion in FTD/ALS gene UNC13A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.06.438573v1?rss=1">
<title>
<![CDATA[
Comprehensive Proteomic Quantification of Bladder Stone Progression in a Cystinuric Mouse Model Using Data-Independent Acquisitions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.06.438573v1?rss=1</link>
<description><![CDATA[
Cystinuria is one of various disorders that cause biomineralization in the urinary system, including bladder stone formation in humans. It is most prevalent in children and adolescents and more aggressive in males. There is no cure, and only limited disease management techniques help to solubilize the stones. Recurrence, even after treatment, occurs frequently. Other than a buildup of cystine, little is known about factors involved in the formation, expansion, and recurrence of these stones. This study sought to define the growth of bladder stones, guided by micro-computed tomography imaging, and to profile dynamic stone proteome changes in a cystinuria mouse model. After bladder stones developed in vivo, they were harvested and separated into four developmental stages (sand, small, medium and large stone), based on their size. Data-dependent and data-independent acquisitions allowed deep profiling of stone proteomics. The proteomic signatures and pathways illustrated major changes as the stones grew. Stones initiate from a small nidus, grow outward, and show major enrichment in ribosomal proteins and factors related to coagulation and platelet degranulation, suggesting a major dysregulation in specific pathways that can be targeted for new therapeutic options.
]]></description>
<dc:creator>Rose, J.</dc:creator>
<dc:creator>Basisty, N.</dc:creator>
<dc:creator>Zee, T.</dc:creator>
<dc:creator>Wehrfritz, C.</dc:creator>
<dc:creator>Bose, N.</dc:creator>
<dc:creator>Desprez, P.-Y.</dc:creator>
<dc:creator>Kapahi, P.</dc:creator>
<dc:creator>Stoller, M.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:date>2021-04-06</dc:date>
<dc:identifier>doi:10.1101/2021.04.06.438573</dc:identifier>
<dc:title><![CDATA[Comprehensive Proteomic Quantification of Bladder Stone Progression in a Cystinuric Mouse Model Using Data-Independent Acquisitions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.06.438722v1?rss=1">
<title>
<![CDATA[
Ligand Strain Energy in Large Library Docking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.06.438722v1?rss=1</link>
<description><![CDATA[
While small molecule internal strain is crucial to molecular docking, using it in evaluating ligand scores has remained elusive. Here, we investigate a technique that calculates strain using relative torsional populations in the Cambridge Structural Database, enabling fast pre-calculation of these energies. In retrospective studies of large docking screens of the dopamine D4 receptor and of AmpC {beta}-lactamase, where close to 600 docking hits were tested experimentally, including such strain energies improved hit rates by preferentially reducing high-scoring decoy molecules that were strained. In a 40 target subset of the DUD-E benchmark, we found two thresholds that usefully distinguished between ligands and decoys: one based on the total strain energy of the small molecules, and one based on the maximum strain allowed for any given torsion within them. Using these criteria, about 75% of the benchmark targets had improved enrichment after strain filtering. Relying on pre-calculated population distributions, this approach is rapid, taking less than 0.04 second to evaluate a conformation on a standard core, making it pragmatic for pre-calculating strain in even ultra-large libraries. Since it is scoring function agnostic, it may be useful to multiple docking approaches; it is openly available at http://tldr.docking.org
]]></description>
<dc:creator>Gu, S.</dc:creator>
<dc:creator>Smith, M. S.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.06.438722</dc:identifier>
<dc:title><![CDATA[Ligand Strain Energy in Large Library Docking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.07.438852v1?rss=1">
<title>
<![CDATA[
Autism risk gene POGZ promotes chromatin accessibility and expression of clustered synaptic genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.07.438852v1?rss=1</link>
<description><![CDATA[
De novo mutations in POGZ, which encodes the chromatin regulator Pogo Transposable Element with ZNF Domain protein, are strongly associated with autism spectrum disorder (ASD). Here we find that in the developing mouse and human brain POGZ binds predominantly euchromatic loci and these are enriched for human neurodevelopmental disorder genes and transposable elements. We profile chromatin accessibility and gene expression in Pogz-/- mice and find that POGZ promotes chromatin accessibility of candidate regulatory elements (REs) and the expression of clustered synaptic genes. We further demonstrate that POGZ forms a nuclear complex and co-occupies loci with HP1{gamma} and ADNP, another high-confidence ASD risk gene. In Pogz+/- mice, Adnp expression is reduced. We postulate that reduced POGZ dosage disrupts cortical function through alterations in the POGZ-ADNP balance which modifies neuronal gene expression.
]]></description>
<dc:creator>Markenscoff, E.</dc:creator>
<dc:creator>Binyameen, F.</dc:creator>
<dc:creator>Whalen, S.</dc:creator>
<dc:creator>Price, J.</dc:creator>
<dc:creator>Lim, K.</dc:creator>
<dc:creator>Catta-Preta, R.</dc:creator>
<dc:creator>Pai, E. L.-L.</dc:creator>
<dc:creator>Mu, X.</dc:creator>
<dc:creator>Xu, D.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Nord, A.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Rubenstein, J. L. R.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438852</dc:identifier>
<dc:title><![CDATA[Autism risk gene POGZ promotes chromatin accessibility and expression of clustered synaptic genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.10.439289v1?rss=1">
<title>
<![CDATA[
An Engineered Contact Lens for Passive and Sustained Release of Lifitegrast, an Anti-Dry Eye Syndrome Drug 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.10.439289v1?rss=1</link>
<description><![CDATA[
Lifitegrast is an FDA-approved drug that inhibits T-cell mediated inflammation associated with dry eye syndrome (DES). Lifitegrast is a potent inhibitor of the interaction between LFA-1 on T-cells and ICAM-1 on endothelial cells at the ocular surface. While effective in treating DES, 5% (81.2 mM) lifitegrast has low drug utilization and elicits off-target effects. Here we engineer contact lenses to release therapeutically-relevant doses of lifitegrast to every tear film for up to 10-hours. Lifitegrast is coupled to the polymer of the soft hydrogel lens via a photolabile (caged) crosslinker. Exposures of the lens to the 400-430 nm wavelengths of indoor daylight excite the caged crosslinker molecules and trigger a bond-cleavage reaction that releases authentic lifitegrast passively to the tear film. The photoproduct of the reaction remains chemically-linked to the polymer of the single-use lens. Our studies show that passive exposures of the lens to indoor light would generate an average of 990 nM lifitegrast to every tear film in a zero-order reaction for up to 10-hours. This concentration exceeds the Kd for the interaction between ICAM-1 and LFA-1 by [~]330-fold and would sustain inhibition of inflammatory responses at the ocular surface. The amount of lifitegrast released from the lens increases during exposures to outdoor sunlight. Over a 10-hour exposure to indoor light, a single lens would release 0.44% of the lifitegrast present in two drops of commercial 5% lifitegrast. Compared to tear-drop approaches, our engineered lenses would sustain the passive delivery of therapeutically-relevant doses of lifitegrast over a longer period, and exhibit improved drug utilization at a lower cost. Our technology could easily be integrated into daily-use contact lenses in order to prevent inflammation at the ocular surface, dry-eye and contact lens-mediated discomfort.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=198 HEIGHT=200 SRC="FIGDIR/small/439289v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Mu, C.</dc:creator>
<dc:creator>Lee, V.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Han, Y.</dc:creator>
<dc:creator>marriott, g.</dc:creator>
<dc:date>2021-04-11</dc:date>
<dc:identifier>doi:10.1101/2021.04.10.439289</dc:identifier>
<dc:title><![CDATA[An Engineered Contact Lens for Passive and Sustained Release of Lifitegrast, an Anti-Dry Eye Syndrome Drug]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.26.441344v1?rss=1">
<title>
<![CDATA[
A Pan-Cancer Census of Dominant Tumor Immune Archetypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.26.441344v1?rss=1</link>
<description><![CDATA[
Cancers display significant heterogeneity with respect to tissue of origin, driver mutations and other features of the surrounding tissue. It is likely that persistent tumors differentially engage inherent patterns-here  Archetypes-of the immune system, to both benefit from a tumor immune microenvironment (TIME) and to disengage tumor-targeting. To discover dominant immune system archetypes, the Immunoprofiler Initiative (IPI) processed 364 individual tumors across 12 cancer types using standardized protocols. Computational clustering of flow cytometry and transcriptomic data obtained from cell sub compartments uncovered archetypes that exist across indications. These Immune composition-based archetypes differentiate tumors based upon unique immune and tumor gene-expression patterns. Archetypes discovered this way also tie closely to well-established classifications of tumor biology. The IPI resource provides a template for understanding cancer immunity as a collection of dominant patterns of immune infiltration and provides a rational path forward to learn how to modulate these patterns to improve therapy.

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]]></description>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Samad, B.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Chew, N. W.</dc:creator>
<dc:creator>Yan, P.</dc:creator>
<dc:creator>Reeder, G. C.</dc:creator>
<dc:creator>Kushnoor, D.</dc:creator>
<dc:creator>Shen, A.</dc:creator>
<dc:creator>Davidson, B.</dc:creator>
<dc:creator>Barczac, A. J.</dc:creator>
<dc:creator>Adkisson, M.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Naser, M.</dc:creator>
<dc:creator>Barry, K. C.</dc:creator>
<dc:creator>Courau, T.</dc:creator>
<dc:creator>Hammoudi, T.</dc:creator>
<dc:creator>Arguello, R. J.</dc:creator>
<dc:creator>Rao, A. A.</dc:creator>
<dc:creator>Olshen, A. B.</dc:creator>
<dc:creator>Immunoprofiler Consortium,</dc:creator>
<dc:creator>Cai, C.</dc:creator>
<dc:creator>Zhan, J.</dc:creator>
<dc:creator>Davis, K. C.</dc:creator>
<dc:creator>Kelley, R. K.</dc:creator>
<dc:creator>Chapman, J. S.</dc:creator>
<dc:creator>Attreya, C. E.</dc:creator>
<dc:creator>Patel, A.</dc:creator>
<dc:creator>Daud, A. I.</dc:creator>
<dc:creator>Ha, P.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:creator>Kratz, J. R.</dc:creator>
<dc:creator>Collisson, E. A.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:creator>Boissonnas, A.</dc:creator>
<dc:creator>Asthana, S.</dc:creator>
<dc:creator>Chan, V.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2021-04-27</dc:date>
<dc:identifier>doi:10.1101/2021.04.26.441344</dc:identifier>
<dc:title><![CDATA[A Pan-Cancer Census of Dominant Tumor Immune Archetypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.28.441880v1?rss=1">
<title>
<![CDATA[
Protracted yet coordinated differentiation of long-lived SARS-CoV-2-specific CD8+ T cells during COVID-19 convalescence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.28.441880v1?rss=1</link>
<description><![CDATA[
CD8+ T cells are important antiviral effectors that can potentiate long-lived immunity against COVID-19, but a detailed characterization of these cells has been hampered by technical challenges. We screened 21 well-characterized, longitudinally-sampled convalescent donors that recovered from mild COVID-19 against a collection of SARS-CoV-2 tetramers, and identified one participant with an immunodominant response against Nuc322-331, a peptide that is conserved in all the SARS-CoV-2 variants-of-concern reported to date. We conducted 38- parameter CyTOF phenotyping on tetramer-identified Nuc322-331-specific CD8+ T cells, and on CD4+ and CD8+ T cells recognizing the entire nucleocapsid and spike proteins from SARS- CoV-2, and took 32 serological measurements on longitudinal specimens from this participant. We discovered a coordination of the Nuc322-331-specific CD8+ T response with both the CD4+ T cell and antibody pillars of adaptive immunity. Nuc322-331-specific CD8+ T cells were predominantly central memory T cells, but continually evolved over a [~]6-month period of convalescence. We observed a slow and progressive decrease in the activation state and polyfunctionality of the Nuc322-331-specific CD8+ T cells, accompanied by an increase in their lymph-node homing and homeostatic proliferation potential. These results suggest that following a typical case of mild COVID-19, SARS-CoV-2-specific CD8+ T cells not only persist but continuously differentiate in a coordinated fashion well into convalescence, into a state characteristic of long-lived, self-renewing memory.
]]></description>
<dc:creator>Ma, T.</dc:creator>
<dc:creator>Ryu, H.</dc:creator>
<dc:creator>McGregor, M.</dc:creator>
<dc:creator>Babcock, B.</dc:creator>
<dc:creator>Neidleman, J.</dc:creator>
<dc:creator>Xie, G.</dc:creator>
<dc:creator>George, A. F.</dc:creator>
<dc:creator>Frouard, J.</dc:creator>
<dc:creator>Murray, V.</dc:creator>
<dc:creator>Gill, G.</dc:creator>
<dc:creator>Ghosn, E.</dc:creator>
<dc:creator>Newell, E.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Roan, N.</dc:creator>
<dc:date>2021-04-29</dc:date>
<dc:identifier>doi:10.1101/2021.04.28.441880</dc:identifier>
<dc:title><![CDATA[Protracted yet coordinated differentiation of long-lived SARS-CoV-2-specific CD8+ T cells during COVID-19 convalescence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.28.441904v1?rss=1">
<title>
<![CDATA[
Regulation of layered T cell tolerance mechanisms by the NR4A family is essential to preserve immune homeostasis and suppress autoimmunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.28.441904v1?rss=1</link>
<description><![CDATA[
The NR4A family of orphan nuclear receptors (Nr4a1-3) plays redundant roles upstream of Foxp3 to establish and maintain Treg identity; deletion of multiple family members in the thymus results in Treg deficiency and a severe inflammatory disease. Consequently, it has been challenging to isolate the functions of this family in other immune cells. Here we take advantage of a competitive bone marrow chimera strategy, coupled with conditional genetic tools, to rescue Treg homeostasis and unmask such functions. Unexpectedly, chimeras harboring Nr4a1-/- Nr4a3-/- (DKO) bone marrow develop autoantibodies and a systemic inflammatory disease despite a replete Treg compartment of largely wild-type origin. This disease differs qualitatively from that seen with Treg-deficiency and is B cell-extrinsic. Negative selection of DKO thymocytes is profoundly impaired in a cell-intrinsic manner. Consistent with escape of self-reactive T cells into the periphery, DKO T cells with functional and phenotypic features of anergy accumulate in chimeric mice. Despite this, DKO T cells exhibit enhanced IL-2 production, implying a cell-intrinsic role for the NR4A family in peripheral T cell tolerance. These studies reveal roles for the NR4A family in multiple layered T cell tolerance mechanisms and demonstrate that each is essential to preserve immune homeostasis.
]]></description>
<dc:creator>Hiwa, R.</dc:creator>
<dc:creator>Nielsen, H. V.</dc:creator>
<dc:creator>Mueller, J. L.</dc:creator>
<dc:creator>Zikherman, J.</dc:creator>
<dc:date>2021-04-29</dc:date>
<dc:identifier>doi:10.1101/2021.04.28.441904</dc:identifier>
<dc:title><![CDATA[Regulation of layered T cell tolerance mechanisms by the NR4A family is essential to preserve immune homeostasis and suppress autoimmunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.29.441889v1?rss=1">
<title>
<![CDATA[
A type I interferon response defines a conserved microglial state required for effective phagocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.29.441889v1?rss=1</link>
<description><![CDATA[
Microglia are brain resident phagocytes that can engulf synaptic components and extracellular matrix as well as whole neurons. However, whether there are unique molecular mechanisms that regulate these distinct phagocytic states is unknown. Here we define a molecularly distinct microglial subset whose function is to engulf neurons in the developing brain. We transcriptomically identified a cluster of Type I interferon (IFN-I) responsive microglia that expanded 20-fold in the postnatal day 5 somatosensory cortex after partial whisker deprivation, a stressor that accelerates neural circuit remodeling. In situ, IFN-I responsive microglia were highly phagocytic and actively engulfed whole neurons. Conditional deletion of IFN-I signaling (Ifnar1fl/fl) in microglia but not neurons resulted in dysmorphic microglia with stalled phagocytosis and an accumulation of neurons with double strand DNA breaks, a marker of cell stress. Conversely, exogenous IFN-I was sufficient to drive neuronal engulfment by microglia and restrict the accumulation of damaged neurons. IFN-I deficient mice had excess excitatory neurons in the developing somatosensory cortex as well as tactile hypersensitivity to whisker stimulation. These data define a molecular mechanism through which microglia engulf neurons during a critical window of brain development. More broadly, they reveal key homeostatic roles of a canonical antiviral signaling pathway in brain development.
]]></description>
<dc:creator>Dorman, L. C.</dc:creator>
<dc:creator>Nguyen, P. T.</dc:creator>
<dc:creator>Escoubas, C. C.</dc:creator>
<dc:creator>Vainchtein, I. D.</dc:creator>
<dc:creator>Xiao, Y.</dc:creator>
<dc:creator>Lidsky, P. V.</dc:creator>
<dc:creator>Wang, E. Y.</dc:creator>
<dc:creator>Taloma, S. E.</dc:creator>
<dc:creator>Nakao-Inoue, H.</dc:creator>
<dc:creator>Rivera, B. M.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Molofsky, A. V.</dc:creator>
<dc:date>2021-04-30</dc:date>
<dc:identifier>doi:10.1101/2021.04.29.441889</dc:identifier>
<dc:title><![CDATA[A type I interferon response defines a conserved microglial state required for effective phagocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.06.443033v1?rss=1">
<title>
<![CDATA[
Active Surveillance Characterizes Human Intratumoral T Cell Exhaustion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.06.443033v1?rss=1</link>
<description><![CDATA[
Intratumoral T cells that might otherwise control tumors are often identified in an  exhausted state, defined by specific epigenetic modifications as well as upregulation of genes such as CD38, CTLA-4 and PD-1. While the term might imply inactivity, there has been little study of this state at the phenotypic level in tumors to understand the extent of their incapacitation. Starting with the observation that T cells move more quickly through mouse tumors as residence time increases and they progress towards exhaustion, we elaborated a non-stimulatory live-biopsy method for real-time study of T cell behaviors within individual patient tumors. Using two-photon microscopy, we studied native CD8 T cells interacting with APCs and with cancer cells in different micro-niches of human tumors, finding that T cell speed was variable by region and by patient, was independent of T cell density and was inversely correlated with local tumor density. Across a range of tumor types, we found a strong relationship between CD8 T cell motility and exhausted T cell state that corresponds to observations made in mouse models where exhausted T cells move faster. While this is a small study, it demonstrates at least two types of T cell dynamic states in individual human tumors and supports the existence of an active program in  exhausted T cells that extends beyond incapacitating them.
]]></description>
<dc:creator>You, R.</dc:creator>
<dc:creator>Artichoker, J.</dc:creator>
<dc:creator>Fries, A.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Reeder, G.</dc:creator>
<dc:creator>Samad, B.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2021-05-07</dc:date>
<dc:identifier>doi:10.1101/2021.05.06.443033</dc:identifier>
<dc:title><![CDATA[Active Surveillance Characterizes Human Intratumoral T Cell Exhaustion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.07.442684v1?rss=1">
<title>
<![CDATA[
Combinatorial immunotherapies overcome MYC-driven immune evasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.07.442684v1?rss=1</link>
<description><![CDATA[
For many human cancers, including triple negative breast cancer (TNBC), a modest number of patients benefit from immune checkpoint inhibitors, and few experience cancer remission1. Expression of programed death-ligand 1 (PD-L1), tumor immune infiltration, or tumor mutation burden have been widely investigated for predicting cancer immunotherapy response1-5. Whether specific oncogenes diminish response to immunotherapy6-10 and whether these effects are reversible remains poorly understood. We predicted that MYC, an oncogene that is frequently overexpressed11,12 and is associated with worse prognosis12, may predict immunotherapy response in patients with TNBC. Here, we report that MYC-elevated TNBCs are resistant to immune checkpoint inhibitors. Using mouse models of TNBC and patient data we report that MYC signaling is associated with low tumor cell PD-L1, low overall immune cell infiltration, and low tumor cell MHC-I expression. Restoring interferon signaling in the tumor reduces MYC expression and increases MHC-I expression. By combining a TLR9 agonist and an agonistic antibody against OX40 with anti-PD-L1, most mice experience complete tumor regression and are protected from new TNBC tumor outgrowth. Our findings demonstrate that MYC-dependent immune evasion is reversible and druggable, and if strategically targeted, may improve outcomes for patients treated with immune checkpoint inhibitors.
]]></description>
<dc:creator>Lee, J. V.</dc:creator>
<dc:creator>Houseley, F.</dc:creator>
<dc:creator>Yau, C.</dc:creator>
<dc:creator>Van de Mark, D.</dc:creator>
<dc:creator>Nakagawa, R.</dc:creator>
<dc:creator>Hemmati, G.</dc:creator>
<dc:creator>Hernandez, G. A.</dc:creator>
<dc:creator>Winkler, J.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Samson, S.</dc:creator>
<dc:creator>Baas, C.</dc:creator>
<dc:creator>Esserman, L. J.</dc:creator>
<dc:creator>Van't Veer, L. J.</dc:creator>
<dc:creator>Rugo, H. S.</dc:creator>
<dc:creator>Matloubian, M.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:date>2021-05-08</dc:date>
<dc:identifier>doi:10.1101/2021.05.07.442684</dc:identifier>
<dc:title><![CDATA[Combinatorial immunotherapies overcome MYC-driven immune evasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.07.443199v1?rss=1">
<title>
<![CDATA[
ATRX promotes heterochromatin formation to protect cells from G-quadruplex DNA-mediated stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.07.443199v1?rss=1</link>
<description><![CDATA[
ATRX is a tumor suppressor that has been associated with protection from DNA replication stress, purportedly through resolution of difficult-to-replicate G-quadruplex (G4) DNA structures. While several studies demonstrate that loss of ATRX sensitizes cells to chemical stabilizers of G4 structures, the molecular function of ATRX at G4 regions during replication remains unknown. Here, we demonstrate that ATRX associates with the MCM replication complex and that loss of ATRX leads to G4 structure accumulation at newly synthesized DNA. We show that both the helicase domain of ATRX and its H3.3 chaperone function are required to protect cells from G4-induced replicative stress. Furthermore, these activities are upstream of heterochromatin formation mediated by the histone methyltransferase, ESET, which is the critical molecular event that protects cells from G4-mediated stress. In support, tumors carrying mutations in either ATRX or ESET show increased mutation burden at G4-enriched DNA sequences. Overall, our study provides new insights into mechanisms by which ATRX promotes genome stability with important implications for understanding impacts of its loss on human disease.
]]></description>
<dc:creator>Teng, Y.-C.</dc:creator>
<dc:creator>Sundaresan, A.</dc:creator>
<dc:creator>O'Hara, R.</dc:creator>
<dc:creator>Gant, V. U.</dc:creator>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Martire, S.</dc:creator>
<dc:creator>Warshaw, J. N.</dc:creator>
<dc:creator>Basu, A.</dc:creator>
<dc:creator>Banaszynski, L.</dc:creator>
<dc:date>2021-05-07</dc:date>
<dc:identifier>doi:10.1101/2021.05.07.443199</dc:identifier>
<dc:title><![CDATA[ATRX promotes heterochromatin formation to protect cells from G-quadruplex DNA-mediated stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.08.443243v1?rss=1">
<title>
<![CDATA[
Assessing chromatin relocalization in 3D using the patient rule induction method 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.08.443243v1?rss=1</link>
<description><![CDATA[
Three dimensional (3D) genome architecture is critical for numerous cellular processes, including transcription, while certain conformation-driven structural alterations are frequently oncogenic. Inferring 3D chromatin configurations has been advanced by the emergence of chromatin conformation capture assays, notably Hi-C, and attendant 3D reconstruction algorithms. These have enhanced understanding of chromatin spatial organization and afforded numerous downstream biological insights. Until recently, comparisons of 3D reconstructions between conditions and/or cell types were limited to prescribed structural features. However, multiMDS, a pioneering approach developed by Rieber and Mahony (2019) that performs joint reconstruction and alignment, enables quantification of all locus-specific differences between paired Hi-C data sets. By subsequently mapping these differences to the linear (1D) genome the identification of relocalization regions is facilitated through use of peak calling in conjunction with continuous wavelet transformation. Here, we seek to refine this approach by performing the search for significant relocalization regions in terms of the 3D structures themselves, thereby retaining the benefits of 3D reconstruction and avoiding limitations associated with the 1D perspective. The search for (extreme) relocalization regions is conducted using the patient rule induction method (PRIM). Considerations surrounding orienting structures with respect to compartmental and principal component axes are discussed, as are approaches to inference and reconstruction accuracy assessment. Illustration makes recourse to comparisons between four different cell types.
]]></description>
<dc:creator>Segal, M. R.</dc:creator>
<dc:date>2021-05-10</dc:date>
<dc:identifier>doi:10.1101/2021.05.08.443243</dc:identifier>
<dc:title><![CDATA[Assessing chromatin relocalization in 3D using the patient rule induction method]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.08.443250v1?rss=1">
<title>
<![CDATA[
Identification of Cancer-Associated Fibroblasts in Glioblastoma and Defining Their Pro-tumoral Effects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.08.443250v1?rss=1</link>
<description><![CDATA[
Despite their identification in some cancers, pro-tumoral cancer-associated fibroblasts (CAFs) were presumed absent in glioblastoma given the lack of brain fibroblasts. Serial trypsinization of primary glioblastoma cultures yielded cells with CAF morphology, CAF transcriptomic profile, and mesenchymal lineage in single-cell RNA-seq. Glioblastoma CAFs were attracted to glioblastoma stem cells (GSCs) and CAFs enriched GSCs. We created a resource of inferred crosstalk by mapping expression of receptors to their cognate ligands, identifying PDGF-{beta} and TGF-{beta} as mediators of GSC effects on CAFs, and osteopontin and hepatocyte growth factor as mediators of CAF-induced GSC enrichment. Glioblastoma CAFs also induced M2 macrophage polarization by producing the EDA fibronectin variant. Glioblastoma CAFs were enriched in the subventricular zone which houses neural stem cells that produce GSCs. Including CAFs in GSC-derived xenografts induced in vivo growth. These findings are among the first to identify glioblastoma CAFs and their GSC interactions, making them an intriguing target.
]]></description>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Rick, J.</dc:creator>
<dc:creator>Joshi, R.</dc:creator>
<dc:creator>Beniwal, A.</dc:creator>
<dc:creator>Spatz, J.</dc:creator>
<dc:creator>Chang, A. C.-C.</dc:creator>
<dc:creator>Nguyen, A. T.</dc:creator>
<dc:creator>Sudhir, S.</dc:creator>
<dc:creator>Chandra, A.</dc:creator>
<dc:creator>Haddad, A.</dc:creator>
<dc:creator>Wadhwa, H.</dc:creator>
<dc:creator>Shah, S. S.</dc:creator>
<dc:creator>Choi, S.</dc:creator>
<dc:creator>Hayes, J. L.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Yagnik, G.</dc:creator>
<dc:creator>Costello, J. F.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:date>2021-05-10</dc:date>
<dc:identifier>doi:10.1101/2021.05.08.443250</dc:identifier>
<dc:title><![CDATA[Identification of Cancer-Associated Fibroblasts in Glioblastoma and Defining Their Pro-tumoral Effects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.08.443268v1?rss=1">
<title>
<![CDATA[
In situ and transcriptomic identification of synapse-associated microglia in the developing zebrafish brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.08.443268v1?rss=1</link>
<description><![CDATA[
Microglia are brain resident macrophages that play vital roles in central nervous system (CNS) development, homeostasis, and pathology. Microglia both remodel synapses and engulf apoptotic cell corpses during development, but whether unique molecular programs regulate these distinct phagocytic functions is unknown. Here we identify a molecularly distinct synapse-associated microglial subset in the zebrafish (Danio rerio). We found that ramified microglia populated synapse-rich regions of the midbrain and hindbrain between 7 and 28 days post fertilization. In contrast, microglia in the optic tectum were ameboid and clustered around neurogenic zones. Using single-cell mRNA sequencing combined with metadata from regional bulk sequencing, we identified synapse-associated microglia (SAMs) that were highly enriched in the hindbrain, expressed known synapse modulating genes as well as novel candidates, and engulfed synaptic proteins. In contrast, neurogenic-associated microglia (NAMs) were enriched in optic tectum, had active cathepsin activity, and preferentially engulfed neuronal corpses. These data yielded a functionally annotated atlas of zebrafish microglia (https://www.annamolofskylab.org/microglia-sequencing). Furthermore, they reveal that molecularly distinct phagocytic programs mediate synaptic remodeling and cell engulfment, and establish zebrafish hindbrain as a model circuit for investigating microglial-synapse interactions.
]]></description>
<dc:creator>Silva, N. J.</dc:creator>
<dc:creator>Dorman, L. C.</dc:creator>
<dc:creator>vainchtein, i.</dc:creator>
<dc:creator>Horneck, N. C.</dc:creator>
<dc:creator>Molofsky, A. V.</dc:creator>
<dc:date>2021-05-10</dc:date>
<dc:identifier>doi:10.1101/2021.05.08.443268</dc:identifier>
<dc:title><![CDATA[In situ and transcriptomic identification of synapse-associated microglia in the developing zebrafish brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.10.443524v1?rss=1">
<title>
<![CDATA[
CryoEM and AI reveal a structure of SARS-CoV-2 Nsp2, a multifunctional protein involved in key host processes. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.10.443524v1?rss=1</link>
<description><![CDATA[
The SARS-CoV-2 protein Nsp2 has been implicated in a wide range of viral processes, but its exact functions, and the structural basis of those functions, remain unknown. Here, we report an atomic model for full-length Nsp2 obtained by combining cryo-electron microscopy with deep learning-based structure prediction from AlphaFold2. The resulting structure reveals a highly-conserved zinc ion-binding site, suggesting a role for Nsp2 in RNA binding. Mapping emerging mutations from variants of SARS-CoV-2 on the resulting structure shows potential host-Nsp2 interaction regions. Using structural analysis together with affinity tagged purification mass spectrometry experiments, we identify Nsp2 mutants that are unable to interact with the actin-nucleation-promoting WASH protein complex or with GIGYF2, an inhibitor of translation initiation and modulator of ribosome-associated quality control. Our work suggests a potential role of Nsp2 in linking viral transcription within the viral replication-transcription complexes (RTC) to the translation initiation of the viral message. Collectively, the structure reported here, combined with mutant interaction mapping, provides a foundation for functional studies of this evolutionary conserved coronavirus protein and may assist future drug design.
]]></description>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Azumaya, C. M.</dc:creator>
<dc:creator>Moritz, M.</dc:creator>
<dc:creator>Pourmal, S.</dc:creator>
<dc:creator>Diallo, A.</dc:creator>
<dc:creator>Merz, G. E.</dc:creator>
<dc:creator>Jang, G. M.</dc:creator>
<dc:creator>Bouhaddou, M.</dc:creator>
<dc:creator>Fossati, A.</dc:creator>
<dc:creator>Brilot, A. F.</dc:creator>
<dc:creator>Diwanji, D.</dc:creator>
<dc:creator>Hernandez, E.</dc:creator>
<dc:creator>Herrera, N.</dc:creator>
<dc:creator>Kratochvil, H. T.</dc:creator>
<dc:creator>Lam, V. L.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Nguyen, H. C.</dc:creator>
<dc:creator>Nowotny, C.</dc:creator>
<dc:creator>Owens, T. W.</dc:creator>
<dc:creator>Peters, J. K.</dc:creator>
<dc:creator>Rizo, A. N.</dc:creator>
<dc:creator>Schulze-Gahmen, U.</dc:creator>
<dc:creator>Smith, A. M.</dc:creator>
<dc:creator>Young, I. D.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Asarnow, D.</dc:creator>
<dc:creator>Billesbolle, C.</dc:creator>
<dc:creator>Campbell, M. G.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Chen, K.-H.</dc:creator>
<dc:creator>Chio, U. S.</dc:creator>
<dc:creator>Dickinson, M. S.</dc:creator>
<dc:creator>Doan, L.</dc:creator>
<dc:creator>Jin, M.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Li, Y.-L.</dc:creator>
<dc:creator>Linossi, E.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Lo, M.</dc:creator>
<dc:creator>Lopez, J.</dc:creator>
<dc:creator>Lopez, K. E.</dc:creator>
<dc:creator>Mancino, A.</dc:creator>
<dc:creator>Moss, F. R.</dc:creator>
<dc:creator>Paul, M. D.</dc:creator>
<dc:creator>Pawar, K. I</dc:creator>
<dc:date>2021-05-12</dc:date>
<dc:identifier>doi:10.1101/2021.05.10.443524</dc:identifier>
<dc:title><![CDATA[CryoEM and AI reveal a structure of SARS-CoV-2 Nsp2, a multifunctional protein involved in key host processes.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.11.443701v1?rss=1">
<title>
<![CDATA[
CRISPR activation and interference screens in primary human T cells decode cytokine regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.11.443701v1?rss=1</link>
<description><![CDATA[
The pathways that regulate cytokine responses in T cells are disrupted in autoimmunity, immune deficiencies, and cancer, and include immunotherapy targets. Systematic discovery of cytokine regulators requires both loss-of-function and gain-of-function studies, which have been challenging in primary human cells. We now have accomplished genome-wide pooled CRISPR activation (CRISPRa) and CRISPR interference (CRISPRi) screens in primary human T cells to map gene networks controlling Interleukin-2 and Interferon-{gamma} production. Arrayed CRISPRa confirmed key hits and enabled multiplexed T cell secretome characterization, revealing reshaped cytokine responses driven by individual regulators. CRISPRa uncovered genes not canonically expressed in T cells, including the transcription factor FOXQ1, whose overexpression promoted the expression of most cytokines, while selectively dampening T helper 2 (Th2) cytokines. Paired CRISPRa and CRISPRi screens reveal signaling components that tune critical immune cell functions, which could inform design of future immunotherapies.
]]></description>
<dc:creator>Schmidt, R.</dc:creator>
<dc:creator>Steinhart, Z.</dc:creator>
<dc:creator>Layeghi, M.</dc:creator>
<dc:creator>Freimer, J. W.</dc:creator>
<dc:creator>Nguyen, V. Q.</dc:creator>
<dc:creator>Blaeschke, F.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2021-05-12</dc:date>
<dc:identifier>doi:10.1101/2021.05.11.443701</dc:identifier>
<dc:title><![CDATA[CRISPR activation and interference screens in primary human T cells decode cytokine regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.12.443888v1?rss=1">
<title>
<![CDATA[
mRNA vaccine-induced SARS-CoV-2-specific T cells recognize B.1.1.7 and B.1.351 variants but differ in longevity and homing properties depending on prior infection status 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.12.443888v1?rss=1</link>
<description><![CDATA[
While mRNA vaccines are proving highly efficacious against SARS-CoV-2, it is important to determine how booster doses and prior infection influence the immune defense they elicit, and whether they protect against variants. Focusing on the T cell response, we conducted a longitudinal study of infection-naive and COVID-19 convalescent donors before vaccination and after their first and second vaccine doses, using a high-parameter CyTOF analysis to phenotype their SARS-CoV-2-specific T cells. Vaccine-elicited spike-specific T cells responded similarly to stimulation by spike epitopes from the ancestral, B.1.1.7 and B.1.351 variant strains, both in terms of cell numbers and phenotypes. In infection-naive individuals, the second dose boosted the quantity and altered the phenotypic properties of SARS-CoV-2-specific T cells, while in convalescents the second dose changed neither. Spike-specific T cells from convalescent vaccinees differed strikingly from those of infection-naive vaccinees, with phenotypic features suggesting superior long-term persistence and ability to home to the respiratory tract including the nasopharynx. These results provide reassurance that vaccine-elicited T cells respond robustly to emerging viral variants, confirm that convalescents may not need a second vaccine dose, and suggest that vaccinated convalescents may have more persistent nasopharynx-homing SARS-CoV-2-specific T cells compared to their infection-naive counterparts.
]]></description>
<dc:creator>Neidleman, J.</dc:creator>
<dc:creator>Luo, X.</dc:creator>
<dc:creator>McGregor, M.</dc:creator>
<dc:creator>Xie, G.</dc:creator>
<dc:creator>Murray, V.</dc:creator>
<dc:creator>Greene, W. C.</dc:creator>
<dc:creator>Lee, S. A.</dc:creator>
<dc:creator>Roan, N. R.</dc:creator>
<dc:date>2021-05-12</dc:date>
<dc:identifier>doi:10.1101/2021.05.12.443888</dc:identifier>
<dc:title><![CDATA[mRNA vaccine-induced SARS-CoV-2-specific T cells recognize B.1.1.7 and B.1.351 variants but differ in longevity and homing properties depending on prior infection status]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.13.444045v1?rss=1">
<title>
<![CDATA[
Molecular diversity and lineage commitment of human interneuron progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.13.444045v1?rss=1</link>
<description><![CDATA[
Cortical interneurons are indispensable for proper function of neocortical circuits. Changes in interneuron development and function are implicated in human disorders, such as autism spectrum disorder and epilepsy. In order to understand human-specific features of cortical development as well as the origins of neurodevelopmental disorders it is crucial to identify the molecular programs underlying human interneuron development and subtype specification. Recent studies have explored gene expression programs underlying mouse interneuron specification and maturation. We applied single-cell RNA sequencing to samples of second trimester human ganglionic eminence and developing cortex to identify molecularly defined subtypes of human interneuron progenitors and immature interneurons. In addition, we integrated this data from the developing human ganglionic eminences and neocortex with single-nucleus RNA-seq of adult cortical interneurons in order to elucidate dynamic molecular changes associated with commitment of progenitors and immature interneurons to mature interneuron subtypes. By comparing our data with published mouse single-cell genomic data, we discover a number of divergent gene expression programs that distinguish human interneuron progenitors from mouse. Moreover, we find that a number of transcription factors expressed during prenatal development become restricted to adult interneuron subtypes in the human but not the mouse, and these adult interneurons express species- and lineage-specific cell adhesion and synaptic genes. Therefore, our study highlights that despite the similarity of main principles of cortical interneuron development and lineage commitment between mouse and human, human interneuron genesis and subtype specification is guided by species-specific gene programs, contributing to human-specific features of cortical inhibitory interneurons.
]]></description>
<dc:creator>Velmeshev, D.</dc:creator>
<dc:creator>Chavali, M.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Bhade, M.</dc:creator>
<dc:creator>Mayer, S.</dc:creator>
<dc:creator>Goyal, N.</dc:creator>
<dc:creator>Alvarado, B.</dc:creator>
<dc:creator>Mancia, W.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Speir, M.</dc:creator>
<dc:creator>Haeussler, M. J.</dc:creator>
<dc:creator>Rowitch, D.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:creator>Huang, E.</dc:creator>
<dc:creator>Paredes, M.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:date>2021-05-14</dc:date>
<dc:identifier>doi:10.1101/2021.05.13.444045</dc:identifier>
<dc:title><![CDATA[Molecular diversity and lineage commitment of human interneuron progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.13.444051v1?rss=1">
<title>
<![CDATA[
When rare meets common: Treatable genetic diseases are enriched in the general psychiatric population 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.13.444051v1?rss=1</link>
<description><![CDATA[
1.1Mental illnesses are one of the biggest contributors to the global disease burden. Despite the increased recognition, diagnosis and ongoing research of mental health disorders, the etiology and underlying molecular mechanisms of these disorders are yet to be fully elucidated. Moreover, despite many treatment options available, a large subset of the psychiatric patient population is non-responsive to standard medications and therapies. There has not been a comprehensive study to date examining the burden and impact of treatable genetic disorders (TGDs) that can present with neuropsychiatric features in psychiatric patient populations. In this study, we test the hypothesis that TGDs that present with psychiatric symptoms are more prevalent within psychiatric patient populations compared to the general population by performing targeted next-generation sequencing (NGS) of 129 genes associated with 108 TGDs in a cohort of 2301 psychiatric patients. In total, 72 putative affected and 293 putative carriers for TGDs were identified, with known or likely pathogenic variants in 78 genes. Despite screening for only 108 genetic disorders, this study showed an approximately four-fold (4.13%) enrichment for genetic disorders within the psychiatric population relative to the estimated 1% cumulative prevalence of all single gene disorders globally. This strongly suggests that the prevalence of these, and most likely all, genetic diseases are greatly underestimated in psychiatric populations. Increasing awareness and ensuring accurate diagnosis of TGDs will open new avenues to targeted treatment for a subset of psychiatric patients.
]]></description>
<dc:creator>Sriretnakumar, V.</dc:creator>
<dc:creator>Harripaul, R.</dc:creator>
<dc:creator>Kennedy, J. L.</dc:creator>
<dc:creator>So, J.</dc:creator>
<dc:date>2021-05-18</dc:date>
<dc:identifier>doi:10.1101/2021.05.13.444051</dc:identifier>
<dc:title><![CDATA[When rare meets common: Treatable genetic diseases are enriched in the general psychiatric population]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.17.444528v1?rss=1">
<title>
<![CDATA[
An Atlas of Cortical Arealization Identifies Dynamic Molecular Signatures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.17.444528v1?rss=1</link>
<description><![CDATA[
The human brain is subdivided into distinct anatomical structures. The neocortex, one of these structures, enables higher-order sensory, associative, and cognitive functions, and in turn encompasses dozens of distinct specialized cortical areas. Early morphogenetic gradients are known to establish an early blueprint for the specification of brain regions and cortical areas. Furthermore, recent studies have uncovered distinct transcriptomic signatures between opposing poles of the developing neocortex1. However, how early, broad developmental patterns result in finer and more discrete spatial differences across the adult human brain remains poorly understood2. Here, we use single-cell RNA-sequencing to profile ten major brain structures and six neocortical areas during peak neurogenesis and early gliogenesis. Our data reveal that distinct cell subtypes are predominantly brain-structure specific. Within the neocortex, we find that even early in the second trimester, a large number of genes are differentially expressed across distinct cortical areas in all cell types, including radial glia, the neural progenitors of the cortex. However, the abundance of areal transcriptomic signatures increases as radial glia differentiate into intermediate progenitor cells and ultimately give rise to excitatory neurons. Using an automated, multiplexed single-molecule fluorescent in situ hybridization (smFISH) approach, we validated the expression pattern of area-specific neuronal genes and also discover that laminar gene expression patterns are highly dynamic across cortical regions. Together, our data suggest that early cortical areal patterning is defined by strong, mutually exclusive frontal and occipital gene expression signatures, with resulting gradients giving rise to the specification of areas between these two poles throughout successive developmental timepoints.
]]></description>
<dc:creator>Bhaduri, A.</dc:creator>
<dc:creator>Sandoval-Espinosa, C.</dc:creator>
<dc:creator>Otero-Garcia, M.</dc:creator>
<dc:creator>Oh, I.</dc:creator>
<dc:creator>Yin, R.</dc:creator>
<dc:creator>Eze, U. C.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Kriegstein, A. R.</dc:creator>
<dc:date>2021-05-17</dc:date>
<dc:identifier>doi:10.1101/2021.05.17.444528</dc:identifier>
<dc:title><![CDATA[An Atlas of Cortical Arealization Identifies Dynamic Molecular Signatures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.17.444554v1?rss=1">
<title>
<![CDATA[
A systematic comparison of fibroblasts derived from postmortem human dura mater versus dermal epithelium for neurodegenerative disease modeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.17.444554v1?rss=1</link>
<description><![CDATA[
Patient-derived cells hold great promise for precision medicine approaches in human health. Fibroblast cells have been a major source of human cells for reprogramming and differentiating into specific cell types for disease modeling. Such cells can be isolated at various stages during life (presymptomatic, symptomatic, and postmortem) and thus can potentially be used to model different phases of disease progression. In certain circumstances, however, tissues are not collected during life and only postmortem tissues are the only available source of fibroblasts. Fibroblasts cultured from postmortem human dura mater of individuals with neurodegenerative diseases have been suggested as a primary source of cells for in vitro modeling of neurodegenerative diseases. Although fibroblast-like cells from human and mouse dura mater have been previously described, their utility for reprogramming and direct differentiation protocols requires further characterization. In this study, cells derived from dermal biopsies performed in living subjects were compared to cells derived from postmortem dura mater. In two instances, we have isolated and compared dermal and dural cell lines from the same subject. Notably, striking differences between the dermis and dura mater-derived cell lines were found. Compared to dermal fibroblasts, postmortem dura mater-derived cells demonstrated different morphology, exhibited slower growth rates, failed to express fibroblast protein markers, and exhibited significant differences in gene expression profiles. In addition, dura mater-derived cells were found to exhibit a high rate of chromosomal abnormalities, particularly in the loss of the Y chromosome. Our study highlights potential limitations of postmortem human dura mater-derived cells for disease modeling, argues for rigorous karyotyping prior to reprograming, and brings into question the identity of dura mater-derived cells as belonging to a fibroblast lineage.
]]></description>
<dc:creator>Argouarch, A. R.</dc:creator>
<dc:creator>Cosme, C. G.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Corrales, C. I.</dc:creator>
<dc:creator>Nana, A. L.</dc:creator>
<dc:creator>Karydas, A. M.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Miller, B.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Kao, A. W.</dc:creator>
<dc:date>2021-05-18</dc:date>
<dc:identifier>doi:10.1101/2021.05.17.444554</dc:identifier>
<dc:title><![CDATA[A systematic comparison of fibroblasts derived from postmortem human dura mater versus dermal epithelium for neurodegenerative disease modeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.24.445507v1?rss=1">
<title>
<![CDATA[
The molecular mechanism of load adaptation by branched actin networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.24.445507v1?rss=1</link>
<description><![CDATA[
Branched actin networks are self-assembling molecular motors that move biological membranes and drive many important cellular processes. Load forces slow the growth and increase the density of these networks, but the molecular mechanisms governing this force response are not well understood. Here we use single-molecule imaging and AFM cantilever deflection to measure how applied forces affect each step in branched actin network assembly. Unexpectedly, force slows the rate of filament nucleation by promoting the interaction of nucleation promoting factors with actin filament ends, limiting branch formation. This inhibition is countered by an even larger force-induced drop in the rate of filament capping, resulting in a shift in the balance between nucleation and capping that increases network density. Remarkably, the force dependence of capping is identical to that of filament elongation because they require the same size gap to appear between the filament and load for insertion. These results provide direct evidence that Brownian Ratchets generate force and govern the load adaptation of branched actin networks.
]]></description>
<dc:creator>Li, T.-D.</dc:creator>
<dc:creator>Bieling, P.</dc:creator>
<dc:creator>Weichsel, J.</dc:creator>
<dc:creator>Mullins, R. D.</dc:creator>
<dc:creator>Fletcher, D. A.</dc:creator>
<dc:date>2021-05-25</dc:date>
<dc:identifier>doi:10.1101/2021.05.24.445507</dc:identifier>
<dc:title><![CDATA[The molecular mechanism of load adaptation by branched actin networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.25.445695v1?rss=1">
<title>
<![CDATA[
Assembly and levels of P-TEFb depend on reversiblephosphorylation of cyclinT1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.25.445695v1?rss=1</link>
<description><![CDATA[
The positive transcription elongation factor b (P-TEFb) is a critical co-activator for transcription of most cellular and viral genes, including those of HIV. While P-TEFb is regulated by 7SK snRNA in proliferating cells, it is absent in quiescent and terminally differentiated cells, which has remained unexplored. In these cells, we found that CycT1 not bound to CDK9 is rapidly degraded. Moreover, productive CycT1:CDK9 interactions require phosphorylation of two threonine residues (Thr143 and Thr149) in CycT1 by PKC. Conversely, PP1 dephosphorylates these sites. Thus, PKC inhibitors or removal of PKC by chronic activation results in P-TEFb disassembly and CycT1 degradation. This finding not only recapitulates P-TEFb depletion in resting CD4+ T cells but also in anergic T cells. Importantly, our studies reveal mechanisms of P-TEFb inactivation underlying T cell quiescence, anergy, and exhaustion as well as proviral latency and terminal differentiation of cells.
]]></description>
<dc:creator>Huang, F.</dc:creator>
<dc:creator>Nguyen, T. N. N.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Ramachandran, R.</dc:creator>
<dc:creator>Cary, D. C.</dc:creator>
<dc:creator>Paculova, H.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Weiss, A.</dc:creator>
<dc:creator>Peterlin, B. M.</dc:creator>
<dc:creator>Fujinaga, K.</dc:creator>
<dc:date>2021-05-25</dc:date>
<dc:identifier>doi:10.1101/2021.05.25.445695</dc:identifier>
<dc:title><![CDATA[Assembly and levels of P-TEFb depend on reversiblephosphorylation of cyclinT1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.27.446019v1?rss=1">
<title>
<![CDATA[
Robust Segmentation of Cellular Ultrastructure on Sparsely Labeled 3D Electron Microscopy Images using Deep Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.27.446019v1?rss=1</link>
<description><![CDATA[
A deeper understanding of the cellular and subcellular organization of tumor cells and their interactions with the tumor microenvironment will shed light on how cancer evolves and guide effective therapy choices. Electron microscopy (EM) images can provide detailed view of the cellular ultrastructure and are being generated at an ever-increasing rate. However, the bottleneck in their analysis is the delineation of the cellular structures to enable interpretable rendering. We have mitigated this limitation by using deep learning, specifically, the ResUNet architecture, to segment cells and subcellular ultrastructure. Our initial prototype focuses on segmenting nuclei and nucleoli in 3D FIB-SEM images of tumor biopsies obtained from patients with metastatic breast and pancreatic cancers. Trained with sparse manual labels, our method results in accurate segmentation of nuclei and nucleoli with best Dice score of 0.99 and 0.98 respectively. This method can be extended to other cellular structures, enabling deeper analysis of inter- and intracellular state and interactions.
]]></description>
<dc:creator>Machireddy, A.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Loftis, K. G.</dc:creator>
<dc:creator>Stoltz, K.</dc:creator>
<dc:creator>Bueno, C. E.</dc:creator>
<dc:creator>Smith, H. R.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:date>2021-05-28</dc:date>
<dc:identifier>doi:10.1101/2021.05.27.446019</dc:identifier>
<dc:title><![CDATA[Robust Segmentation of Cellular Ultrastructure on Sparsely Labeled 3D Electron Microscopy Images using Deep Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.27.446027v1?rss=1">
<title>
<![CDATA[
Mechanical Forces during Lymph Node Expansion Govern Fibroblastic Reticular Network Remodeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.27.446027v1?rss=1</link>
<description><![CDATA[
Emergent physical properties of tissues are not readily understood by reductionist studies of their constituent cells. Here, we show molecular signals controlling cellular physical properties, collectively determining tissue mechanics of lymph nodes, an immunologically-relevant, adult mammalian tissue. Lymph nodes paradoxically maintain robust tissue architecture in homeostasis yet are continually poised for extensive tissue expansion upon immune challenge. We find that following immune challenge, cytoskeletal mechanics of a cellular meshwork of fibroblasts determine tissue tension independently of extracellular matrix scaffolds. We determine that CLEC-2/podoplanin signalling regulates the cell surface mechanics of fibroblasts, permitting cell elongation and interdigitation through expedited access to plasma membrane reservoirs. Increased tissue tension through the stromal meshwork gates the initiation of fibroblast proliferation, restoring homeostatic cellular ratios and tissue structure through expansion.
]]></description>
<dc:creator>Horsnell, H. L.</dc:creator>
<dc:creator>Tetley, R. J.</dc:creator>
<dc:creator>de Belly, H.</dc:creator>
<dc:creator>Makris, S.</dc:creator>
<dc:creator>Benjamin, A. C.</dc:creator>
<dc:creator>Paluch, E.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Acton, S. E.</dc:creator>
<dc:date>2021-05-27</dc:date>
<dc:identifier>doi:10.1101/2021.05.27.446027</dc:identifier>
<dc:title><![CDATA[Mechanical Forces during Lymph Node Expansion Govern Fibroblastic Reticular Network Remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.01.444518v1?rss=1">
<title>
<![CDATA[
Glucocorticoid receptor collaborates with pioneer factors and AP-1 to execute genome-wide regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.01.444518v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWThe glucocorticoid receptor (GR) regulates transcription through binding to specific DNA motifs, particularly at enhancers. While the motif to which it binds is constant across cell types, GR has cell type-specific binding at genomic loci, resulting in regulation of different genes. The presence of other bound transcription factors (TFs) is hypothesized to strongly influence where GR binds. Here, we addressed the roles of other TFs in the glucocorticoid response by comparing changes in GR binding and nascent transcription at promoters and distal candidate cis-regulatory elements (CCREs) in two distinct human cancer cell types. We found that after glucocorticoid treatment, GR binds to thousands of genomic loci that are primarily outside of promoter regions and are potentially enhancers. The majority of these GR binding sites are cell-type specific, and they are associated with pioneer factor binding. A small fraction of GR occupied regions (GORs) displayed increased bidirectional nascent transcription, which is a characteristic of many active enhancers, after glucocorticoid treatment. Non-promoter GORs with increased transcription were specifically enriched for AP-1 binding prior to glucocorticoid treatment. These results support a model of transcriptional regulation in which multiple classes of TFs are required. The pioneer factors increase chromatin accessibility, facilitating the binding of GR and additional factors. AP-1 binding poises a fraction of accessible sites to be rapidly transcribed upon glucocorticoid-induced GR binding. The coordinated activity of multiple TFs then results in cell type-specific changes in gene expression. We anticipate that many models of inducible gene expression also require multiple distinct TFs that act at multiple steps of transcriptional regulation.
]]></description>
<dc:creator>Wissink, E. M.</dc:creator>
<dc:creator>Martinez, D. M.</dc:creator>
<dc:creator>Ehmsen, K. T.</dc:creator>
<dc:creator>Yamamoto, K. R.</dc:creator>
<dc:creator>Lis, J. T.</dc:creator>
<dc:date>2021-06-01</dc:date>
<dc:identifier>doi:10.1101/2021.06.01.444518</dc:identifier>
<dc:title><![CDATA[Glucocorticoid receptor collaborates with pioneer factors and AP-1 to execute genome-wide regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.01.446640v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 infection studies in lung organoids identify TSPAN8 as novel mediator 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.01.446640v1?rss=1</link>
<description><![CDATA[
SARS coronavirus-2 (SARS-CoV-2) is causing a global pandemic with large variation in COVID-19 disease spectrum. SARS-CoV-2 infection requires host receptor ACE2 on lung epithelium, but epithelial underpinnings of variation are largely unknown. We capitalized on comprehensive organoid assays to report remarkable variation in SARS-CoV-2 infection rates of lung organoids from different subjects. Tropism is highest for TUBA- and MUC5AC-positive organoid cells, but levels of TUBA-, MUC5A-, or ACE2-positive cells do not predict infection rate. We identify surface molecule Tetraspanin 8 (TSPAN8) as novel mediator of SARS-CoV-2 infection, which is not downregulated by this specific virus. TSPAN8 levels, prior to infection, strongly correlate with infection rate and TSPAN8-blocking antibodies diminish SARS-CoV-2 infection. We propose TSPAN8 as novel functional biomarker and potential therapeutic target for COVID-19.
]]></description>
<dc:creator>Hysenaj, L.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:creator>Kulhanek, K. R.</dc:creator>
<dc:creator>Gbenedio, O.</dc:creator>
<dc:creator>Rodriguez, L.</dc:creator>
<dc:creator>Shen, A.</dc:creator>
<dc:creator>Lone, J.-C.</dc:creator>
<dc:creator>Lupin-Jimenez, L. C.</dc:creator>
<dc:creator>Bonser, L.</dc:creator>
<dc:creator>Serwas, N. K.</dc:creator>
<dc:creator>Bahl, K.</dc:creator>
<dc:creator>Mick, E.</dc:creator>
<dc:creator>Li, J. Z.</dc:creator>
<dc:creator>Ding, V.</dc:creator>
<dc:creator>Matsumoto, S.</dc:creator>
<dc:creator>Maishan, M. l.</dc:creator>
<dc:creator>Fragiadakis, G.</dc:creator>
<dc:creator>Jablons, D. M.</dc:creator>
<dc:creator>Langelier, C. R.</dc:creator>
<dc:creator>Matthay, M.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:creator>Krummel, M.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Erle, D. M.</dc:creator>
<dc:creator>Kratz, J. R.</dc:creator>
<dc:creator>Roose, J. P.</dc:creator>
<dc:date>2021-06-02</dc:date>
<dc:identifier>doi:10.1101/2021.06.01.446640</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 infection studies in lung organoids identify TSPAN8 as novel mediator]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.02.446741v1?rss=1">
<title>
<![CDATA[
High activity and high functional connectivity are mutually exclusive in resting state zebrafish and human brains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.02.446741v1?rss=1</link>
<description><![CDATA[
Active neurons impact cell types with which they are functionally connected. Both activity and functional connectivity are heterogeneous across the brain, but the nature of their relationship is not known. Here we employ brain-wide calcium imaging at cellular resolution in larval zebrafish to record spontaneous activity of >12,000 neurons in the forebrain. By classifying their activity and functional connectivity into three levels (high, medium, low), we find that highly active neurons have low functional connections and highly connected neurons are of low activity. Intriguingly, deploying the same analytical methods on functional magnetic resonance imaging (fMRI) data from the resting state human brain, we uncover a similar relationship between activity and functional connectivity, that is, regions of high activity are non-overlapping with those of high connectivity. These findings reveal a previously unknown and evolutionarily conserved brain organizational principle that have implications for understanding disease states and designing artificial neuronal networks.
]]></description>
<dc:creator>Zarei, M.</dc:creator>
<dc:creator>Xie, D.</dc:creator>
<dc:creator>Jiang, F.</dc:creator>
<dc:creator>Bagirov, A.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:date>2021-06-02</dc:date>
<dc:identifier>doi:10.1101/2021.06.02.446741</dc:identifier>
<dc:title><![CDATA[High activity and high functional connectivity are mutually exclusive in resting state zebrafish and human brains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.04.447088v1?rss=1">
<title>
<![CDATA[
Single-cell RNA-sequencing reveals widespread personalized, context-specific gene expression regulation in immune cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.04.447088v1?rss=1</link>
<description><![CDATA[
Gene expression and its regulation can be context-dependent. To dissect this, using samples from 120 individuals, we single-cell RNA-sequenced 1.3M peripheral blood mononuclear cells exposed to three different pathogens at two time points or left unexposed. This revealed thousands of cell type-specific expression changes (eQTLs) and pathogen-induced expression changes (response QTLs) that are influenced by genetic variation. In monocytes, the strongest responder to pathogen stimulations, genetics also affected co-expression of 71.4% of these eQTL genes. For example, the pathogen recognition receptor CLEC12A showed many such co-expression interactions, but only in monocytes after 3h pathogen stimulation. Further analysis linked this to interferon-regulating transcription factors, a finding that we recapitulated in an independent cohort of patients with systemic lupus erythematosus, a condition characterized by increased interferon activity. Altogether, this study highlights the importance of context for gaining a better understanding of the mechanisms of gene regulation in health and disease.
]]></description>
<dc:creator>Oelen, R.</dc:creator>
<dc:creator>de Vries, D. H.</dc:creator>
<dc:creator>Brugge, H.</dc:creator>
<dc:creator>Gordon, G.</dc:creator>
<dc:creator>Vochteloo, M.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Westra, H.-J.</dc:creator>
<dc:creator>Franke, L.</dc:creator>
<dc:creator>van der Wijst, M. G. P.</dc:creator>
<dc:date>2021-06-06</dc:date>
<dc:identifier>doi:10.1101/2021.06.04.447088</dc:identifier>
<dc:title><![CDATA[Single-cell RNA-sequencing reveals widespread personalized, context-specific gene expression regulation in immune cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.06.446826v1?rss=1">
<title>
<![CDATA[
Evolution of enhanced innate immune evasion by the SARS-CoV-2 B.1.1.7 UK variant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.06.446826v1?rss=1</link>
<description><![CDATA[
Emergence of SARS-CoV-2 variants, including the globally successful B.1.1.7 lineage, suggests viral adaptations to host selective pressures resulting in more efficient transmission. Although much effort has focused on Spike adaptation for viral entry and adaptive immune escape, B.1.1.7 mutations outside Spike likely contribute to enhance transmission. Here we used unbiased abundance proteomics, phosphoproteomics, mRNA sequencing and viral replication assays to show that B.1.1.7 isolates more effectively suppress host innate immune responses in airway epithelial cells. We found that B.1.1.7 isolates have dramatically increased subgenomic RNA and protein levels of Orf9b and Orf6, both known innate immune antagonists. Expression of Orf9b alone suppressed the innate immune response through interaction with TOM70, a mitochondrial protein required for RNA sensing adaptor MAVS activation, and Orf9b binding and activity was regulated via phosphorylation. We conclude that B.1.1.7 has evolved beyond the Spike coding region to more effectively antagonise host innate immune responses through upregulation of specific subgenomic RNA synthesis and increased protein expression of key innate immune antagonists. We propose that more effective innate immune antagonism increases the likelihood of successful B.1.1.7 transmission, and may increase in vivo replication and duration of infection.
]]></description>
<dc:creator>Thorne, L. G.</dc:creator>
<dc:creator>Bouhaddou, M.</dc:creator>
<dc:creator>Reuschl, A.-K.</dc:creator>
<dc:creator>Zuliani-Alvarez, L.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Pelin, A.</dc:creator>
<dc:creator>Batra, J.</dc:creator>
<dc:creator>Whelan, M. V. X.</dc:creator>
<dc:creator>Ummadi, M.</dc:creator>
<dc:creator>Roic, A.</dc:creator>
<dc:creator>Turner, J.</dc:creator>
<dc:creator>Obernier, K.</dc:creator>
<dc:creator>Braberg, H.</dc:creator>
<dc:creator>Soucheray, M.</dc:creator>
<dc:creator>Richards, A. L.</dc:creator>
<dc:creator>Chen, K.-H.</dc:creator>
<dc:creator>Harjai, B.</dc:creator>
<dc:creator>Memon, D.</dc:creator>
<dc:creator>Hosmillo, M.</dc:creator>
<dc:creator>Hiatt, J.</dc:creator>
<dc:creator>Jahun, A. S.</dc:creator>
<dc:creator>Goodfellow, I. G.</dc:creator>
<dc:creator>Noursadeghi, M.</dc:creator>
<dc:creator>Fabius, J.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:creator>Jura, N.</dc:creator>
<dc:creator>Verba, K. A.</dc:creator>
<dc:creator>Beltrao, P.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Garcia-Sastre, A.</dc:creator>
<dc:creator>Jolly, C.</dc:creator>
<dc:creator>Towers, G. J.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:date>2021-06-07</dc:date>
<dc:identifier>doi:10.1101/2021.06.06.446826</dc:identifier>
<dc:title><![CDATA[Evolution of enhanced innate immune evasion by the SARS-CoV-2 B.1.1.7 UK variant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.07.447473v1?rss=1">
<title>
<![CDATA[
Viral Evasion of the Integrated Stress Response Through Antagonistic eIF2-P Mimicry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.07.447473v1?rss=1</link>
<description><![CDATA[
Viral infection triggers activation of the integrated stress response (ISR). In response to viral double-stranded RNA (dsRNA), RNA-activated protein kinase (PKR) phosphorylates the translation initiation factor eIF2, converting it from a translation initiator into a potent translation inhibitor and this restricts the synthesis of viral proteins. Phosphorylated eIF2 (eIF2-P) inhibits translation by binding to eIF2s dedicated, heterodecameric nucleotide exchange factor eIF2B and conformationally inactivating it. We show that the NSs protein of Sandfly Fever Sicilian virus (SFSV) allows the virus to evade the ISR. Mechanistically, NSs tightly binds to eIF2B (KD = 43 nM), blocks eIF2-P binding, and rescues eIF2B GEF activity. Cryo-EM structures demonstrate that SFSV NSs and eIF2-P directly compete, with the primary NSs contacts to eIF2B mediated by five  aromatic fingers. NSs binding preserves eIF2B activity by maintaining eIF2Bs conformation in its active A-State.
]]></description>
<dc:creator>Schoof, M.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Cogan, J. Z.</dc:creator>
<dc:creator>Lawrence, R.</dc:creator>
<dc:creator>Boone, M.</dc:creator>
<dc:creator>Wuerth, J. D.</dc:creator>
<dc:creator>Frost, A.</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:date>2021-06-08</dc:date>
<dc:identifier>doi:10.1101/2021.06.07.447473</dc:identifier>
<dc:title><![CDATA[Viral Evasion of the Integrated Stress Response Through Antagonistic eIF2-P Mimicry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.09.447796v1?rss=1">
<title>
<![CDATA[
Allele-specific gene editing rescues pathology in a human model of Charcot-Marie-Tooth disease type 2E 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.09.447796v1?rss=1</link>
<description><![CDATA[
Many neuromuscular disorders are caused by dominant missense mutations that lead to dominant-negative or gain-of-function pathology. This category of disease is challenging to address via drug treatment or gene augmentation therapy because these strategies may not eliminate the effects of the mutant protein or RNA. Thus, effective treatments are severely lacking for these dominant diseases, which often cause severe disability or death. The targeted inactivation of dominant disease alleles by gene editing is a promising approach with the potential to completely remove the cause of pathology with a single treatment. Here, we demonstrate that allele-specific CRISPR gene editing in a human model of axonal Charcot-Marie-Tooth (CMT) disease rescues pathology caused by a dominant missense mutation in the neurofilament light chain gene (NEFL, CMT type 2E). We utilized a rapid and efficient method for generating spinal motor neurons from human induced pluripotent stem cells (iPSCs) derived from a patient with CMT2E. Diseased motor neurons recapitulated known pathologic phenotypes at early time points of differentiation, including aberrant accumulation of neurofilament light chain protein in neuronal cell bodies. We selectively inactivated the disease NEFL allele in patient iPSCs using Cas9 enzymes to introduce a frameshift at the pathogenic N98S mutation. Motor neurons carrying this allele-specific frameshift demonstrated an amelioration of the disease phenotype comparable to that seen in an isogenic control with precise correction of the mutation. Our results validate allele-specific gene editing as a therapeutic approach for CMT2E and as a promising strategy to silence dominant mutations in any gene for which heterozygous loss-of-function is well tolerated. This highlights the potential for gene editing as a therapy for currently untreatable dominant neurologic diseases.
]]></description>
<dc:creator>Feliciano, C. M.</dc:creator>
<dc:creator>Wu, K.</dc:creator>
<dc:creator>Watry, H. L.</dc:creator>
<dc:creator>Marley, C. B. E.</dc:creator>
<dc:creator>Ramadoss, G. N.</dc:creator>
<dc:creator>Ghanim, H. Y.</dc:creator>
<dc:creator>Liu, A. Z.</dc:creator>
<dc:creator>Zholudeva, L. V.</dc:creator>
<dc:creator>McDevitt, T. C.</dc:creator>
<dc:creator>Saporta, M. A.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:creator>Judge, L. M.</dc:creator>
<dc:date>2021-06-09</dc:date>
<dc:identifier>doi:10.1101/2021.06.09.447796</dc:identifier>
<dc:title><![CDATA[Allele-specific gene editing rescues pathology in a human model of Charcot-Marie-Tooth disease type 2E]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.10.447947v1?rss=1">
<title>
<![CDATA[
Expansion of RNA sequence diversity and RNA editing rates throughout human cortical development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.10.447947v1?rss=1</link>
<description><![CDATA[
Post-transcriptional modifications by RNA editing are essential for neurodevelopment, yet their developmental and regulatory features remain poorly resolved. We constructed a full temporal view of base-specific RNA editing in the developing human cortex, from early progenitors through fully mature cells found in the adult brain. Developmental regulation of RNA editing is characterized by an increase in editing rates for more than 10,000 selective editing sites, shifting between mid-fetal development and infancy, and a massive expansion of RNA hyper-editing sites that amass in the cortex through postnatal development into advanced age. These sites occur disproportionally in 3UTRs of essential neurodevelopmental genes. These profiles are preserved in non-human primate and murine models, illustrating evolutionary conserved regulation of RNA editing in mammalian cortical development. RNA editing levels are commonly genetically regulated (editing quantitative trait loci, edQTLs) consistently across development or predominantly during prenatal or postnatal periods. Both consistent and temporal-predominant edQTLs co-localize with risk loci associated with neurological traits and disorders, including attention deficit hyperactivity disorder, schizophrenia, and sleep disorders. These findings expand the repertoire of highly regulated RNA editing sites in the brain and provide insights of how epitranscriptional sequence diversity by RNA editing contributes to neurodevelopment.
]]></description>
<dc:creator>Cuddleston, R.</dc:creator>
<dc:creator>Sloofman, L.</dc:creator>
<dc:creator>Liang, L.</dc:creator>
<dc:creator>Mossotto, E.</dc:creator>
<dc:creator>Fan, X.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Sestan, N.</dc:creator>
<dc:creator>Devlin, B.</dc:creator>
<dc:creator>Roeder, K.</dc:creator>
<dc:creator>Buxbaum, J. D.</dc:creator>
<dc:creator>Sanders, S.</dc:creator>
<dc:creator>Breen, M. S.</dc:creator>
<dc:date>2021-06-10</dc:date>
<dc:identifier>doi:10.1101/2021.06.10.447947</dc:identifier>
<dc:title><![CDATA[Expansion of RNA sequence diversity and RNA editing rates throughout human cortical development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.16.447601v1?rss=1">
<title>
<![CDATA[
Allosteric Inhibition of a Vesicular Glutamate Transporter by an Isoform-Specific Antibody 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.16.447601v1?rss=1</link>
<description><![CDATA[
The role of glutamate in excitatory neurotransmission depends on its transport into synaptic vesicles by the vesicular glutamate transporters (VGLUTs). The three VGLUT isoforms exhibit a complementary distribution in the nervous system and the knockout of each produces severe, pleiotropic neurological effects. However, the available pharmacology lacks sensitivity and specificity, limiting the analysis of both transport mechanism and physiological role. To develop new molecular probes for the VGLUTs, we raised six mouse monoclonal antibodies to VGLUT2. All six bind to a structured region of VGLUT2, five to the luminal face and one to the cytosolic. Two are specific to VGLUT2 whereas the other four bind to both VGLUT1 and 2; none detect VGLUT3. Antibody 8E11 recognizes an epitope spanning the three extracellular loops in the C-domain that explains the recognition of both VGLUT1 and 2 but not VGLUT3. 8E11 also inhibits both glutamate transport and the VGLUT-associated chloride conductance. Since the antibody binds outside the substrate recognition site, it acts allosterically to inhibit function presumably by restricting conformational changes. The isoform specificity also shows that allosteric inhibition provides a mechanism to distinguish between closely related transporters.
]]></description>
<dc:creator>Eriksen, J.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Stroud, R. M.</dc:creator>
<dc:creator>Edwards, R. H.</dc:creator>
<dc:date>2021-06-16</dc:date>
<dc:identifier>doi:10.1101/2021.06.16.447601</dc:identifier>
<dc:title><![CDATA[Allosteric Inhibition of a Vesicular Glutamate Transporter by an Isoform-Specific Antibody]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.17.448569v1?rss=1">
<title>
<![CDATA[
CUB domain containing protein 1 (CDPC1) is a target for radioligand therapy in castration resistant prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.17.448569v1?rss=1</link>
<description><![CDATA[
PurposeRadioligand therapy (RLT) is relatively unexplored in metastatic castration resistant prostate cancer (mCRPC), with much of the focus having been on bone seeking radionuclides and PSMA-directed RLT. Herein, we evaluated if CUB domain containing protein 1 (CDCP1) can be exploited to treat mCRPC with RLT, particularly for subsets like small cell neuroendocrine prostate cancer (SCNC) that would not be expected to respond to current options.

Experimental DesignCDCP1 mRNA levels were evaluated in the RNA-seq data from 119 recent mCRPC biopsies. Protein expression was assessed in twelve SCNC and adenocarcinoma patient derived xenografts. Saturation binding assays were performed with 4A06, a recombinant human antibody that targets the CDCP1 ectodomain. The feasibility of imaging and treating mCRPC in vivo was tested with 89Zr-4A06 and 177Lu-4A06.

ResultsCDCP1 mRNA expression was observed in over 90% of mCRPC biopsies, including SCNC and in adenocarcinoma with low FOLH1 (PSMA) levels. A modest anticorrelation was observed between CDCP1 and PTEN. Overall survival was not significantly different based on CDCP1 mRNA levels, regardless of PTEN status. Full length and/or cleaved CDCP1 was expressed in ten of twelve PDX samples. Bmax values of ~22,000 and ~6,200 fmol/mg were calculated for two human prostate cancer cell lines. Five prostate cancer models were readily detected in vivo with 89Zr-4A06. 177Lu-4A06 significantly suppressed the growth of DU145 tumors compared to control.

ConclusionsThe antitumor data and the overexpression of CDCP1 reported herein provide the first evidence promoting CDCP1 directed RLT as a treatment strategy for mCRPC.

Statement of Translational RelevanceNew targets for RLT are needed to address the subset of mCRPC that cannot be treated with bone seeking radionuclides or PSMA directed RLT. We report herein the first data credentialing CDCP1 as a target for mCRPC, in both adenocarcinoma and neuroendocrine subtypes. Combined with low expression in normal human tissues, these data provide a compelling scientific rationale for testing CDCP1 directed RLT clinically in mCRPC patients alone or in combination with other systemic therapies.
]]></description>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Trepka, k.</dc:creator>
<dc:creator>Wang, Y.-h.</dc:creator>
<dc:creator>Chopra, S.</dc:creator>
<dc:creator>Hooshdaran, N.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Zhuo, J.</dc:creator>
<dc:creator>Lim, S.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Egusa, E.</dc:creator>
<dc:creator>Zhang, L. K.</dc:creator>
<dc:creator>Foye, A.</dc:creator>
<dc:creator>Chou, J.</dc:creator>
<dc:creator>Feng, F. Y.</dc:creator>
<dc:creator>Small, E. J.</dc:creator>
<dc:creator>Evans, M. J.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:creator>Aggarwal, R.</dc:creator>
<dc:date>2021-06-17</dc:date>
<dc:identifier>doi:10.1101/2021.06.17.448569</dc:identifier>
<dc:title><![CDATA[CUB domain containing protein 1 (CDPC1) is a target for radioligand therapy in castration resistant prostate cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.23.449623v1?rss=1">
<title>
<![CDATA[
Modular, Cascade-like Transcriptional Program of Regeneration in Stentor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.23.449623v1?rss=1</link>
<description><![CDATA[
The giant ciliate Stentor coeruleus is a classical model system for studying regeneration and morphogenesis at the level of a single cell. The anterior of the cell is marked by an array of cilia, known as the oral apparatus, which can be induced to shed and regenerate in a series of reproducible morphological steps, previously shown to require transcription. If a cell is cut in half, each half will regenerate an intact cell, including a new oral apparatus in the posterior half. We used RNAseq to assay the dynamic changes in Stentors transcriptome during regeneration, after both oral apparatus shedding and bisection, allowing us to identify distinct temporal waves of gene expression including kinases, RNA binding proteins, centriole biogenesis factors, and orthologs of human ciliopathy genes implicated in Meckel and Joubert syndromes. By comparing transcriptional profiles of different regeneration events in the same species, we were able to identify distinct modules of gene expression corresponding to oral apparatus regeneration, posterior holdfast regeneration, and recovery after wounding. By measuring gene expression in cells in which translation is blocked, we show that the sequential waves of gene expression involve a cascade mechanism in which later waves of expression are triggered by translation products of early-expressed genes. Among the early-expressed genes, we identified an E2F transcription factor and the conserved RNA binding protein Pumilio as potential regulators of regeneration based on the expression pattern of their predicted target genes. RNAi mediated knockdown experiments indicate that Pumilio is required for regenerating oral structures of the correct size. We show that E2F is involved in the completion of regeneration but is dispensable for earlier steps. This work allows us to classify regeneration genes into groups based on their potential role for regeneration in distinct cell regeneration paradigms, and provides insight into how a single cell can coordinate complex morphogenetic pathways to regenerate missing structures.
]]></description>
<dc:creator>Sood, P.</dc:creator>
<dc:creator>Lin, A.</dc:creator>
<dc:creator>McGillivary, R.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2021-06-23</dc:date>
<dc:identifier>doi:10.1101/2021.06.23.449623</dc:identifier>
<dc:title><![CDATA[Modular, Cascade-like Transcriptional Program of Regeneration in Stentor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.23.449636v1?rss=1">
<title>
<![CDATA[
Aberrant FGF signaling promotes granule neuron precursor expansion in SHH subgroup infantile medulloblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.23.449636v1?rss=1</link>
<description><![CDATA[
Mutations in Sonic Hedgehog (SHH) signaling pathway genes, e.g., Suppressor of Fused (SUFU), drive granule neuron precursors (GNP) to form medulloblastomas (MBSHH). However, how different molecular lesions in the Shh pathway drive transformation is frequently unclear, and SUFU mutations in the cerebellum seem distinct. In this study, we show that fibroblast growth factor 5 (FGF5) signaling is integral for many infantile MBSHH cases and that FGF5 expression is uniquely upregulated in infantile MBSHH tumors. Similarly, mice lacking SUFU (Sufu-cKO) ectopically express Fgf5 specifically along the secondary fissure where GNPs harbor preneoplastic lesions and show that FGFR signaling is also ectopically activated in this region. Treatment with an FGFR antagonist rescues the severe GNP hyperplasia and restores cerebellar architecture. Thus, direct inhibition of FGF signaling may be a promising and novel therapeutic candidate for infantile MBSHH.
]]></description>
<dc:creator>Yabut, O. R.</dc:creator>
<dc:creator>Gomez, H.</dc:creator>
<dc:creator>Arela, J.</dc:creator>
<dc:creator>Castillo, J. G.</dc:creator>
<dc:creator>Ngo, T.</dc:creator>
<dc:creator>Pleasure, S. J.</dc:creator>
<dc:date>2021-06-24</dc:date>
<dc:identifier>doi:10.1101/2021.06.23.449636</dc:identifier>
<dc:title><![CDATA[Aberrant FGF signaling promotes granule neuron precursor expansion in SHH subgroup infantile medulloblastoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.25.449993v1?rss=1">
<title>
<![CDATA[
An acetyl-click screening platform identifies a small molecule inhibitor of Histone Acetyltransferase 1 (HAT1) with anti-tumor activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.25.449993v1?rss=1</link>
<description><![CDATA[
HAT1 is a central regulator of chromatin synthesis that acetylates nascent histone H3:H4 tetramers in the cytoplasm. It may have a role in cancer metabolism by linking cytoplasmic production of acetyl-CoA to nuclear acetyl flux. This is because the HAT1 di-acetylation mark is not propagated in chromatin and instead is de-acetylated after nascent histone insertion into chromatin. Thus, HAT1 likely provides a nuclear source of free acetate that may be recycled to acetyl-CoA for nuclear acetylation reactions. Correspondingly, suppression of HAT1 protein expression impairs tumor growth. To ascertain whether targeting HAT1 is a viable anti-cancer treatment strategy we sought to identify small molecule inhibitors of HAT1. We developed a high-throughput HAT1 acetyl-click assay to facilitate drug discovery and enzymology. Screening of small molecules computationally predicted to bind the active site led to the discovery of multiple riboflavin analogs that inhibited HAT1 enzymatic activity by competing with acetyl-CoA binding. These hits were refined by synthesis and testing over 70 analogs, which yielded structure-activity relationships. The isoalloxazine core was required for enzymatic inhibition, whereas modifications of the ribityl sidechain improved enzymatic potency and cellular growth suppression. These efforts resulted in a lead compound (JG-2016) that suppressed growth of human cancer cells lines in vitro and impaired tumor growth in vivo. This is the first report of a small molecule inhibitor of the HAT1 enzyme complex and represents a step towards targeting this pathway for cancer therapy.
]]></description>
<dc:creator>Gruber, J. J.</dc:creator>
<dc:creator>Rangarajan, A.</dc:creator>
<dc:creator>Chou, T.</dc:creator>
<dc:creator>Geller, B. S.</dc:creator>
<dc:creator>Banuelos, S.</dc:creator>
<dc:creator>Greenhouse, R.</dc:creator>
<dc:creator>Snyder, M.</dc:creator>
<dc:creator>Lipchik, A. M.</dc:creator>
<dc:date>2021-06-26</dc:date>
<dc:identifier>doi:10.1101/2021.06.25.449993</dc:identifier>
<dc:title><![CDATA[An acetyl-click screening platform identifies a small molecule inhibitor of Histone Acetyltransferase 1 (HAT1) with anti-tumor activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.27.450111v1?rss=1">
<title>
<![CDATA[
Origins and timing of emerging lesions in advanced renal cell carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.27.450111v1?rss=1</link>
<description><![CDATA[
PurposeRenal cell carcinoma (RCC) with venous tumor thrombus (VTT) arising from the primary tumor occurs in 4-10% of cases and is associated with advanced disease. RCC with VTT and distant metastasis represents a unique clinical entity, and provides opportunities to examine the origins and relative timing of tumor lesion emergence and to identify molecular correlates with disease state.

Experimental DesignWe performed genomic and evolutionary analyses on 16 RCC patients with VTT, with eight also having metastases, using multi-region exome and RNA sequencing.

ResultsNo genomic alterations were specifically associated with the VTT or metastasis lesions; each tumor had multiple hallmark driver alterations, consistent with advanced disease state. We found that 21% (3/14) of clear-cell RCC cases could be assigned a previously defined "evolutionary subtype". Somatic mutation signatures were largely consistent with previously established RCC signatures, and showed low heterogeneity across regions of each tumor. Mismatch repair and homologous recombination ("BRCA-ness") deficiency signatures consistently co-occurred across most tumors, suggesting a pervasive role for intracellular DNA damage in RCC and the potential for related treatment strategies. Phylogenetic timing analysis of metastatic cases suggested that in most tumors, metastases branched from the primary tumor prior to formation of VTT and in some cases before diversification of the primary tumor. Both VTT and the earliest metastases were predicted to emerge many years prior to diagnosis. Transcriptional landscape analysis identified key differences distinguishing each lesion type from primary tumor: VTT upregulated TNF signaling and associated inflammatory pathways, whereas metastases upregulated MTOR signaling.

ConclusionsOur results provide a map of how RCC tumors can evolve, with metastatic clones typically emerging early in RCC development and taking hold via MTOR signaling, and later formation of VTT via local inflammatory processes.

Statement of Translational RelevanceRenal cell carcinoma (RCC) is a deadly and relatively common malignancy, which often presents as or progresses to metastatic disease. We used multi-region sequencing of RCC patients with venous tumor thrombus (VTT) and metastasis to ask how and when new lesions arise from the primary tumor, and what genomic factors contribute to their spread. Phylogenetic analysis of patients with VTT and co-presenting metastases suggested that in most cases, the VTT and metastases derive from distinct tumor clones. Moreover, metastatic clones often appear many years prior to diagnosis. We found that local TNF inflammation may contribute to VTT formation, whereas MTOR signaling is associated with metastases. Our study sheds light on the relationship of VTT and metastases, suggests therapeutic and biomarker strategies for RCC, and points to the need for early detection studies in RCC to better understand when metastases emerge and to identify at-risk patients.
]]></description>
<dc:creator>Wallace, A.</dc:creator>
<dc:creator>Porten, S.</dc:creator>
<dc:creator>Lo, A. A.</dc:creator>
<dc:creator>Oreper, D.</dc:creator>
<dc:creator>Lounsbury, N.</dc:creator>
<dc:creator>Havnar, C.</dc:creator>
<dc:creator>Pechuan-Jorge, X.</dc:creator>
<dc:creator>Zill, O.</dc:creator>
<dc:creator>Meng, M.</dc:creator>
<dc:date>2021-06-28</dc:date>
<dc:identifier>doi:10.1101/2021.06.27.450111</dc:identifier>
<dc:title><![CDATA[Origins and timing of emerging lesions in advanced renal cell carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.29.450179v1?rss=1">
<title>
<![CDATA[
Innate, translation-dependent silencing of an invasive transposon in Arabidopsis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.29.450179v1?rss=1</link>
<description><![CDATA[
Co-evolution between hosts and parasites genomes shapes diverse pathways of acquired immunity based on silencing small (s)RNAs. In plants, sRNAs cause heterochromatinization, sequence-degeneration and, ultimately, loss-of-autonomy of most transposable elements (TEs). Recognition of newly-invasive plant TEs, by contrast, involves an innate antiviral-like silencing response. To investigate this responses activation, we studied the single-copy element EVADE (EVD), one of few representatives of the large Ty1/Copia family able to proliferate in Arabidopsis when epigenetically-reactivated. In Ty1/Copia-elements, a short subgenomic mRNA (shGAG) provides the necessary excess of structural GAG protein over the catalytic components encoded by the full-length genomic flGAG-POL. We show here that the predominant cytosolic distribution of shGAG strongly favors its translation over mostly-nuclear flGAG-POL, during which an unusually intense ribosomal stalling event coincides precisely with the starting-point of sRNA production exclusively on shGAG. mRNA breakage occurring at this starting-point yields unconventional 5OH RNA fragments that evade RNA-quality-control and concomitantly likely stimulate RNA-DEPENDENT-RNA-POLYMERASE-6 (RDR6) to initiate sRNA production. This hitherto-unrecognized "translation-dependent silencing" (TdS) is independent of codon-usage or GC-content and is not observed on TE remnants populating the Arabidopsis genome, consistent with their poor association, if any, with polysomes. We propose that TdS forms a primal defense against de novo invasive TEs that underlies their associated sRNA patterns.
]]></description>
<dc:creator>Oberlin, S.</dc:creator>
<dc:creator>Rajeswaran, R.</dc:creator>
<dc:creator>Trasser, M.</dc:creator>
<dc:creator>Barragan-Borrero, V.</dc:creator>
<dc:creator>Schon, M. A.</dc:creator>
<dc:creator>Plotnikova, A.</dc:creator>
<dc:creator>Loncsek, L.</dc:creator>
<dc:creator>Nodine, M. D.</dc:creator>
<dc:creator>Mari-Ordonez, A.</dc:creator>
<dc:creator>Voinnet, O.</dc:creator>
<dc:date>2021-06-29</dc:date>
<dc:identifier>doi:10.1101/2021.06.29.450179</dc:identifier>
<dc:title><![CDATA[Innate, translation-dependent silencing of an invasive transposon in Arabidopsis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.30.450566v1?rss=1">
<title>
<![CDATA[
Short homology-directed repair using optimized Cas9 in the pathogen Cryptococcus neoformans  enables rapid gene deletion and tagging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.30.450566v1?rss=1</link>
<description><![CDATA[
Cryptococcus neoformans, the most common cause of fungal meningitis, is a basidiomycete haploid budding yeast with a complete sexual cycle. Genome modification by homologous recombination is feasible using biolistic transformation and long homology arms, but the method is arduous and unreliable. Recently, multiple groups have reported the use of CRISPR-Cas9 as an alternative to biolistics, but long homology arms are still necessary, limiting the utility of this method. Since the S. pyogenes Cas9 derivatives used in prior studies were not optimized for expression in C. neoformans, we designed, synthesized, and tested a fully C. neoformans-optimized Cas9. We found that a Cas9 harboring only common C. neoformans codons and a consensus C. neoformans intron together with a TEF1 promoter and terminator and a nuclear localization signal (C. neoformans-optimized CAS9 or "CnoCAS9") reliably enabled genome editing in the widely-used KN99 C. neoformans strain. Furthermore, editing was accomplished using donors harboring short (50 bp) homology arms attached to marker DNAs produced with synthetic oligonucleotides and PCR amplification. We also demonstrated that prior stable integration of CnoCAS9 further enhances both transformation and homologous recombination efficiency; importantly, this manipulation does not impact virulence in animals. We also implemented a universal tagging module harboring a codon-optimized fluorescent protein (mNeonGreen) and a tandem Calmodulin Binding Peptide-2X FLAG Tag that allows for both localization and purification studies of proteins for which the corresponding genes are modified by short homology-directed recombination. These tools enable short-homology genome engineering in C. neoformans.
]]></description>
<dc:creator>Huang, M. Y.</dc:creator>
<dc:creator>Joshi, M. B.</dc:creator>
<dc:creator>Boucher, M. J.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Loza, L. C.</dc:creator>
<dc:creator>Gaylord, E. A.</dc:creator>
<dc:creator>Doering, T. L.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:date>2021-06-30</dc:date>
<dc:identifier>doi:10.1101/2021.06.30.450566</dc:identifier>
<dc:title><![CDATA[Short homology-directed repair using optimized Cas9 in the pathogen Cryptococcus neoformans  enables rapid gene deletion and tagging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.04.451050v1?rss=1">
<title>
<![CDATA[
Fly Cell Atlas: a single-cell transcriptomic atlas of the adult fruit fly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.04.451050v1?rss=1</link>
<description><![CDATA[
The ability to obtain single cell transcriptomes for stable cell types and dynamic cell states is ushering in a new era for biology. We created the Tabula Drosophilae, a single cell atlas of the adult fruit fly which includes 580k cells from 15 individually dissected sexed tissues as well as the entire head and body. Over 100 researchers from the fly community contributed annotations to >250 distinct cell types across all tissues. We provide an in-depth analysis of cell type-related gene signatures and transcription factor markers, as well as sexual dimorphism, across the whole animal. Analysis of common cell types that are shared between tissues, such as blood and muscle cells, allowed the discovery of rare cell types and tissue-specific subtypes. This atlas provides a valuable resource for the entire Drosophila community and serves as a comprehensive reference to study genetic perturbations and disease models at single cell resolution.
]]></description>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Janssens, J.</dc:creator>
<dc:creator>De Waegeneer, M.</dc:creator>
<dc:creator>Kolluru, S. S.</dc:creator>
<dc:creator>Davie, K.</dc:creator>
<dc:creator>Gardeux, V.</dc:creator>
<dc:creator>Sealens, W.</dc:creator>
<dc:creator>David, F.</dc:creator>
<dc:creator>Brbic, M.</dc:creator>
<dc:creator>Leskovec, J.</dc:creator>
<dc:creator>McLaughlin, C.</dc:creator>
<dc:creator>Xie, Q.</dc:creator>
<dc:creator>Jones, R.</dc:creator>
<dc:creator>Brueckner, K.</dc:creator>
<dc:creator>Shim, J.</dc:creator>
<dc:creator>Tattikota, S. G.</dc:creator>
<dc:creator>Schnorrer, F.</dc:creator>
<dc:creator>Rust, K.</dc:creator>
<dc:creator>Nystul, T.</dc:creator>
<dc:creator>Carvalho-Santos, Z.</dc:creator>
<dc:creator>Ribeiro, C.</dc:creator>
<dc:creator>Pal, S.</dc:creator>
<dc:creator>Przytycka, T. M.</dc:creator>
<dc:creator>Allen, A.</dc:creator>
<dc:creator>Goodwin, S.</dc:creator>
<dc:creator>Berry, C.</dc:creator>
<dc:creator>Fuller, M. T.</dc:creator>
<dc:creator>White-Cooper, H.</dc:creator>
<dc:creator>Matunis, E.</dc:creator>
<dc:creator>DiNardo, S.</dc:creator>
<dc:creator>Galenza, A.</dc:creator>
<dc:creator>O'Brien, L. E.</dc:creator>
<dc:creator>Dow, J. A. T.</dc:creator>
<dc:creator>FCA Consortium,</dc:creator>
<dc:creator>Jasper, H.</dc:creator>
<dc:creator>Oliver, B.</dc:creator>
<dc:creator>Perrimon, N.</dc:creator>
<dc:creator>Deplancke, B.</dc:creator>
<dc:creator>Quake, S.</dc:creator>
<dc:creator>Luo, L.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:date>2021-07-05</dc:date>
<dc:identifier>doi:10.1101/2021.07.04.451050</dc:identifier>
<dc:title><![CDATA[Fly Cell Atlas: a single-cell transcriptomic atlas of the adult fruit fly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.07.451502v1?rss=1">
<title>
<![CDATA[
Holistic Characterization of Tumor Monocyte-to-Macrophage Differentiation Integrates Distinct Immune Phenotypes in Kidney Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.07.451502v1?rss=1</link>
<description><![CDATA[
The tumor immune microenvironment (TIME) is commonly infiltrated by diverse collections of myeloid cells. Yet, the complexity of myeloid cell identity and plasticity has challenged efforts to define bona fide populations and determine their connections to T cell function and their relation to patient outcome. Here we leverage single-cell RNA-sequencing (scRNA-seq) analysis of several mouse and human tumors and find that monocyte-macrophage diversity is characterized by a combination of conserved lineage states as well as transcriptional programs accessed along the differentiation trajectory. Using mouse models, we also find that tumor monocyte-to-macrophage progression is profoundly tied to regulatory T cell (Treg) abundance. Importantly, in human kidney cancer, heterogeneity in macrophage accumulation and myeloid composition corresponded to variance in, not only Treg density, but also the quality of infiltrating CD8+ T cells. In this way, holistic analysis of monocyte-to-macrophage differentiation creates a framework for critically different immune states in kidney tumors.
]]></description>
<dc:creator>Mujal, A. M.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Rao, A. A.</dc:creator>
<dc:creator>Binnewies, M.</dc:creator>
<dc:creator>Samad, B.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Boissonnas, A.</dc:creator>
<dc:creator>Pollack, J. L.</dc:creator>
<dc:creator>Arguello, R. J.</dc:creator>
<dc:creator>Ruhland, M. K.</dc:creator>
<dc:creator>Barry, K. C.</dc:creator>
<dc:creator>Chan, V.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2021-07-08</dc:date>
<dc:identifier>doi:10.1101/2021.07.07.451502</dc:identifier>
<dc:title><![CDATA[Holistic Characterization of Tumor Monocyte-to-Macrophage Differentiation Integrates Distinct Immune Phenotypes in Kidney Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.10.451667v1?rss=1">
<title>
<![CDATA[
High-resolution estimates of crossover and noncrossover recombination from a captive baboon colony 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.10.451667v1?rss=1</link>
<description><![CDATA[
Homologous recombination has been extensively studied in humans and a handful of model organisms. Much less is known about recombination in other species, including non-human primates. Here we present a study of crossovers and non-crossover (NCO) recombination in olive baboons (Papio anubis) from two pedigrees containing a total of 20 paternal and 17 maternal meioses, and compare these results to linkage-disequlibrium (LD) based recombination estimates from 36 unrelated olive baboons. We demonstrate how crossovers, combined with LD-based recombination estimates, can be used to identify genome assembly errors. We also quantify sex-specific differences in recombination rates, including elevated male crossover and reduced female crossover rates near telomeres. Finally, we add to the increasing body of evidence suggesting that while most NCO recombination tracts in mammals are short (e.g., < 500 bp), there are a non-negligible fraction of longer (e.g., > 1 Kb) NCO tracts. We fit a mixture-of-two-geometric distributions model to the NCO tract length distribution and estimate that >99% of all NCO tracts are very short (mean 24 bp), but the remaining tracts can be quite long (mean 11 Kb). A single geometric distribution model for NCO tract lengths is incompatible with the data, suggesting that LD-based methods for estimating NCO recombination rates that make this assumption may need to be modified.

SignificanceMost homologous recombination events are noncrossovers (NCO), but little is known about NCO conversion tract lengths. Here we utilize whole-genome sequence data from large baboon pedigrees to estimate the NCO tract length distribution and to study other aspects of recombination.
]]></description>
<dc:creator>Wall, J.</dc:creator>
<dc:creator>Robinson, J. A.</dc:creator>
<dc:creator>Cox, L. A.</dc:creator>
<dc:date>2021-07-12</dc:date>
<dc:identifier>doi:10.1101/2021.07.10.451667</dc:identifier>
<dc:title><![CDATA[High-resolution estimates of crossover and noncrossover recombination from a captive baboon colony]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.11.451980v1?rss=1">
<title>
<![CDATA[
Pooled screening of CAR T cells identifies non-native signaling domains for next- generation immunotherapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.11.451980v1?rss=1</link>
<description><![CDATA[
Chimeric antigen receptors (CARs) repurpose natural signaling components to retarget T cells to refractory cancers, but have shown limited efficacy against solid tumors. Here, we introduce  CAR Pooling, a multiplexed approach to rapidly identify CAR designs with clinical potential. Forty CARs with diverse immune costimulatory domains were assessed in pooled assays for their ability to stimulate critical T cell effector functions during repetitive stimulation that mimics long-term tumor antigen exposure. Several non-native domains from the TNF receptor family exhibited enhanced proliferation (CD40) or cytotoxicity (BAFF-R and TACI) relative to clinical benchmarks, and fell into distinct states of memory, cytotoxicity, and metabolism. BAFF-R CAR T cells were enriched for a highly cytotoxic and NK-cell-like innate phenotype previously associated with positive clinical outcomes.  CAR Pooling enables efficient exploration of how CAR design affects cell activity and can be applied to optimize receptors across a range of applications and cell types.
]]></description>
<dc:creator>Goodman, D. B.</dc:creator>
<dc:creator>Azimi, C. S.</dc:creator>
<dc:creator>Kearns, K.</dc:creator>
<dc:creator>Garakani, K.</dc:creator>
<dc:creator>Garcia, J.</dc:creator>
<dc:creator>Patel, N.</dc:creator>
<dc:creator>Hwang, B.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Park, E.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Bluestone, J.</dc:creator>
<dc:creator>Roybal, K.</dc:creator>
<dc:date>2021-07-12</dc:date>
<dc:identifier>doi:10.1101/2021.07.11.451980</dc:identifier>
<dc:title><![CDATA[Pooled screening of CAR T cells identifies non-native signaling domains for next- generation immunotherapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.15.452440v1?rss=1">
<title>
<![CDATA[
Functional and Structural Segregation of Overlapping Helices in HIV-1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.15.452440v1?rss=1</link>
<description><![CDATA[
Overlapping coding regions balance selective forces between multiple genes. One possible division of nucleotide sequence is that the predominant selective force on a particular nucleotide can be attributed to just one gene. While this arrangement has been observed in regions in which one gene is structured and the other is disordered, we sought to explore how overlapping genes balance constraints when both protein products are structured over the same sequence. We use a combination of sequence analysis, functional assays and selection experiments to examine an overlapped region in HIV-1 that encodes helical regions in both Env and Rev. We find that functional segregation occurs even in this overlap, with each protein spacing its functional residues in a manner that allows a mutable non-binding face of one helix to encode important functional residues on a charged face in the other helix. Additionally, our experiments reveal novel and critical functional residues in Env and have implications for the therapeutic targeting of HIV-1.
]]></description>
<dc:creator>Safari, M.</dc:creator>
<dc:creator>Jayaraman, B.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Fernandes, J. D.</dc:creator>
<dc:creator>Frankel, A. D.</dc:creator>
<dc:date>2021-07-15</dc:date>
<dc:identifier>doi:10.1101/2021.07.15.452440</dc:identifier>
<dc:title><![CDATA[Functional and Structural Segregation of Overlapping Helices in HIV-1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.15.452565v1?rss=1">
<title>
<![CDATA[
Data-independent acquisition protease-multiplexing enables increased proteome sequence coverage across multiple fragmentation modes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.15.452565v1?rss=1</link>
<description><![CDATA[
The use of multiple proteases has been shown to increase protein sequence coverage in proteomics experiments, however due to the additional analysis time required, it has not been widely adapted in routine data-dependent acquisition (DDA) proteomic workflows. Alternatively, data-independent acquisition (DIA) has the potential to analyze multiplexed samples from different protease digests, but has been primarily optimized for fragmenting tryptic peptides. Here we evaluate a DIA multiplexing approach that combines three proteolytic digests (Trypsin, AspN, and GluC) into a single sample. We first optimize data acquisition conditions for each protease individually with both the canonical DIA fragmentation mode (beam type CID), as well as resonance excitation CID, to determine optimal consensus conditions across proteases. Next, we demonstrate that application of these conditions to a protease-multiplexed sample of human peptides results in similar protein identifications and quantitative performance as compared to trypsin alone, but enables up to a 63% increase in peptide detections, and a 27% increase non-redundant amino acid detections. Importantly, this resulted in 100% sequence coverage for numerous proteins, suggesting the utility of this approach in applications where sequence coverage is critical, such as proteoform analysis.
]]></description>
<dc:creator>Richards, A. L.</dc:creator>
<dc:creator>Chen, K.-H.</dc:creator>
<dc:creator>Wilburn, D. B.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Polacco, B.</dc:creator>
<dc:creator>Searle, B. C.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:date>2021-07-15</dc:date>
<dc:identifier>doi:10.1101/2021.07.15.452565</dc:identifier>
<dc:title><![CDATA[Data-independent acquisition protease-multiplexing enables increased proteome sequence coverage across multiple fragmentation modes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.18.452833v1?rss=1">
<title>
<![CDATA[
Deep neural language modeling enables functional protein generation across families 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.18.452833v1?rss=1</link>
<description><![CDATA[
Bypassing natures evolutionary trajectory, de novo protein generation--defined as creating artificial protein sequences from scratch--could enable breakthrough solutions for biomedical and environmental challenges. Viewing amino acid sequences as a language, we demonstrate that a deep learning-based language model can generate functional artificial protein sequences across families, akin to generating grammatically and semantically correct natural language sentences on diverse topics. Our protein language model is trained by simply learning to predict the next amino acid for over 280 million protein sequences from thousands of protein families, without biophysical or coevolutionary modeling. We experimentally evaluate model-generated artificial proteins on five distinct antibacterial lysozyme families. Artificial proteins show similar activities and catalytic efficiencies as representative natural lysozymes, including hen egg white lysozyme, while reaching as low as 44% identity to any known naturally-evolved protein. The X-ray crystal structure of an enzymatically active artificial protein recapitulates the conserved fold and positioning of active site residues found in natural proteins. We demonstrate our language models ability to be adapted to different protein families by accurately predicting the functionality of artificial chorismate mutase and malate dehydrogenase proteins. These results indicate that neural language models successfully perform de novo protein generation across protein families and may prove to be a tool to shortcut evolution.
]]></description>
<dc:creator>Madani, A.</dc:creator>
<dc:creator>Krause, B.</dc:creator>
<dc:creator>Greene, E. R.</dc:creator>
<dc:creator>Subramanian, S.</dc:creator>
<dc:creator>Mohr, B. P.</dc:creator>
<dc:creator>Holton, J. M.</dc:creator>
<dc:creator>Olmos, J. L.</dc:creator>
<dc:creator>Xiong, C.</dc:creator>
<dc:creator>Sun, Z. Z.</dc:creator>
<dc:creator>Socher, R.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Naik, N.</dc:creator>
<dc:date>2021-07-18</dc:date>
<dc:identifier>doi:10.1101/2021.07.18.452833</dc:identifier>
<dc:title><![CDATA[Deep neural language modeling enables functional protein generation across families]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.20.453090v1?rss=1">
<title>
<![CDATA[
Single-cell genomics reveals region-specific developmental trajectories underlying neuronal diversity in the prenatal human hypothalamus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.20.453090v1?rss=1</link>
<description><![CDATA[
The development and diversity of neuronal subtypes in the human hypothalamus has been insufficiently characterized. We sequenced the transcriptomes of 40,927 cells from the prenatal human hypothalamus spanning from 6 to 25 gestational weeks and 25,424 mature neurons in regions of the adult human hypothalamus, revealing a temporal trajectory from proliferative stem cell populations to mature neurons and glia. Developing hypothalamic neurons followed branching trajectories leading to 170 transcriptionally distinct neuronal subtypes in ten hypothalamic nuclei in the adult. The uniqueness of hypothalamic neuronal lineages was examined developmentally by comparing excitatory lineages present in cortex and inhibitory lineages in ganglionic eminence from the same individuals, revealing both distinct and shared drivers of neuronal maturation across the human forebrain. Cross-species comparisons to the mouse hypothalamus identified human-specific POMC populations expressing unique combinations of transcription factors and neuropeptides. These results provide the first comprehensive transcriptomic view of human hypothalamus development at cellular resolution.

One-Sentence SummaryUsing single-cell genomics, we reconstructed the developmental lineages by which precursor populations give rise to 170 distinct neuronal subtypes in the human hypothalamus.
]]></description>
<dc:creator>Herb, B.</dc:creator>
<dc:creator>Glover, H. J.</dc:creator>
<dc:creator>Bhaduri, A.</dc:creator>
<dc:creator>Casella, A. M.</dc:creator>
<dc:creator>Bale, T. L.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:creator>Doege, C.</dc:creator>
<dc:creator>Ament, S. A.</dc:creator>
<dc:date>2021-07-20</dc:date>
<dc:identifier>doi:10.1101/2021.07.20.453090</dc:identifier>
<dc:title><![CDATA[Single-cell genomics reveals region-specific developmental trajectories underlying neuronal diversity in the prenatal human hypothalamus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.29.453595v1?rss=1">
<title>
<![CDATA[
Internet of Things Architecture for High Throughput Biology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.29.453595v1?rss=1</link>
<description><![CDATA[
The Internet of Things (IoT) provides a simple framework to easily control online devices. IoT is now a commonplace tool used by technology companies, but it is rarely used in biology experiments. IoT can benefit research through alarm notifications, automation, and the real-time monitoring of experiments. We developed and implemented an IoT architecture to control biological devices used in experiments. We developed our own electrophysiology, microscopy, and microfluidic devices so that may be controlled through a unified IoT architecture. The system allows each device to be monitored and controlled through an online web tool. We present our IoT architecture so other labs may replicate it for their own experiments.
]]></description>
<dc:creator>Parks, D. F.</dc:creator>
<dc:creator>Voitiuk, K.</dc:creator>
<dc:creator>Geng, J.</dc:creator>
<dc:creator>Elliott, M. A. T.</dc:creator>
<dc:creator>Keefe, M. G.</dc:creator>
<dc:creator>Jung, E. A.</dc:creator>
<dc:creator>Robbins, A.</dc:creator>
<dc:creator>Baudin, P. V.</dc:creator>
<dc:creator>Ly, V. T.</dc:creator>
<dc:creator>Hawthorne, N.</dc:creator>
<dc:creator>Yong, D.</dc:creator>
<dc:creator>Sanso, S. E.</dc:creator>
<dc:creator>Rezaee, N.</dc:creator>
<dc:creator>Sevetson, J. L.</dc:creator>
<dc:creator>Seiler, S. T.</dc:creator>
<dc:creator>Currie, R.</dc:creator>
<dc:creator>Hengen, K. B.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Salama, S. R.</dc:creator>
<dc:creator>Teodorescu, M.</dc:creator>
<dc:creator>Haussler, D.</dc:creator>
<dc:date>2021-08-01</dc:date>
<dc:identifier>doi:10.1101/2021.07.29.453595</dc:identifier>
<dc:title><![CDATA[Internet of Things Architecture for High Throughput Biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.30.454497v1?rss=1">
<title>
<![CDATA[
Xenotransplantation of porcine progenitor cells in an epileptic California sea lion (Zalophus californianus) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.30.454497v1?rss=1</link>
<description><![CDATA[
BackgroundDomoic acid (DA) is a naturally occurring neurotoxin harmful to marine animals and humans. California sea lions exposed to DA in prey during algal blooms along the Pacific coast exhibit significant neurological symptoms, including epilepsy with hippocampal atrophy.

ObservationsHere we describe a xenotransplantation procedure to deliver interneuron progenitor cells into the damaged hippocampus of an epileptic sea lion with suspected DA toxicosis. The sea lion has had no evidence of seizures following the procedure, and clinical measures of well-being including weight and feeding habits have stabilized.

LessonsThese preliminary results suggest xenotransplantation has improved the quality-of-life (QOL) for this animal and holds tremendous therapeutic promise.
]]></description>
<dc:creator>Simeone, C. A.</dc:creator>
<dc:creator>Andrews, J. P.</dc:creator>
<dc:creator>Johnson, S. P.</dc:creator>
<dc:creator>Casalia, M.</dc:creator>
<dc:creator>Kochanski, R.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Cameron, D.</dc:creator>
<dc:creator>Dennison, S.</dc:creator>
<dc:creator>Inglis, B.</dc:creator>
<dc:creator>Scott, G.</dc:creator>
<dc:creator>Kruse-Elliott, K.</dc:creator>
<dc:creator>Okonski, F. F.</dc:creator>
<dc:creator>Calvo, E.</dc:creator>
<dc:creator>Goulet, K.</dc:creator>
<dc:creator>Robles, D.</dc:creator>
<dc:creator>Griffin-Stence, A.</dc:creator>
<dc:creator>Kuiper, E.</dc:creator>
<dc:creator>Krasovec, L.</dc:creator>
<dc:creator>Field, C. L.</dc:creator>
<dc:creator>Hoard, V. F.</dc:creator>
<dc:creator>Baraban, S. C.</dc:creator>
<dc:date>2021-07-31</dc:date>
<dc:identifier>doi:10.1101/2021.07.30.454497</dc:identifier>
<dc:title><![CDATA[Xenotransplantation of porcine progenitor cells in an epileptic California sea lion (Zalophus californianus)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.05.455250v1?rss=1">
<title>
<![CDATA[
Interleukin-33 coordinates a microglial phagocytic response and limits corticothalamic excitability and seizure susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.05.455250v1?rss=1</link>
<description><![CDATA[
Microglia are key remodelers of neuronal synapses during brain development, but the mechanisms that regulate this process and its ultimate impact on neural circuit function are not well defined. We previously identified the IL-1 family cytokine Interleukin-33 (IL-33) as a novel mediator of microglial synapse remodeling. Here we define the phagocytic program induced in microglia in response to IL-33. We find that IL-33 markedly alters the microglial enhancer landscape and exposes AP-1 transcription factor sites that promote target gene expression. We identify the scavenger receptor MARCO and the pattern recognition receptor TLR2 as downstream mediators of IL-33 dependent synapse engulfment. Conditional deletion of IL-33 in the CNS or its receptor on microglia results in increased numbers of excitatory synapses in the corticothalamic circuit and spontaneous epileptiform activity as well as increased seizure susceptibility by early adulthood. These findings define novel mechanisms through which IL-33 coordinates experience-dependent synaptic refinement to restrict hyperexcitability in the developing brain.
]]></description>
<dc:creator>Han, R. T.</dc:creator>
<dc:creator>Vainchtein, I. D.</dc:creator>
<dc:creator>Schlachetzki, J. C. M.</dc:creator>
<dc:creator>Cho, F. S.</dc:creator>
<dc:creator>Dorman, L. C.</dc:creator>
<dc:creator>Johung, T.</dc:creator>
<dc:creator>Ahn, E.</dc:creator>
<dc:creator>Barron, J. T.</dc:creator>
<dc:creator>Inoue, H. N.</dc:creator>
<dc:creator>Joshi, A.</dc:creator>
<dc:creator>Molofsky, A. B.</dc:creator>
<dc:creator>Glass, C. K.</dc:creator>
<dc:creator>Paz, J. T.</dc:creator>
<dc:creator>Molofsky, A. V.</dc:creator>
<dc:date>2021-08-06</dc:date>
<dc:identifier>doi:10.1101/2021.08.05.455250</dc:identifier>
<dc:title><![CDATA[Interleukin-33 coordinates a microglial phagocytic response and limits corticothalamic excitability and seizure susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.06.455445v1?rss=1">
<title>
<![CDATA[
Visual modulation of spectrotemporal receptive fields in mouse auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.06.455445v1?rss=1</link>
<description><![CDATA[
Recent studies have established significant anatomical and functional connections between visual areas and primary auditory cortex (A1), which may be important for perceptual processes such as communication and spatial perception. However, much remains unknown about the microcircuit structure of these interactions, including how visual context may affect different cell types across cortical layers, each with diverse responses to sound. The present study examined activity in putative excitatory and inhibitory neurons across cortical layers of A1 in awake male and female mice during auditory, visual, and audiovisual stimulation. We observed a subpopulation of A1 neurons responsive to visual stimuli alone, which were overwhelmingly found in the deep cortical layers and included both excitatory and inhibitory cells. Other neurons for which responses to sound were modulated by visual context were similarly excitatory or inhibitory but were less concentrated within the deepest cortical layers. Important distinctions in visual context sensitivity were observed among different spike rate and timing responses to sound. Spike rate responses were themselves heterogeneous, with stronger responses evoked by sound alone at stimulus onset, but greater sensitivity to visual context by sustained firing activity following transient onset responses. Minimal overlap was observed between units with visual-modulated firing rate responses and spectrotemporal receptive fields (STRFs) which are sensitive to both spike rate and timing changes. Together, our results suggest visual information in A1 is predominantly carried by deep layer inputs and influences sound encoding across cortical layers, and that these influences independently impact qualitatively distinct responses to sound.

Significance statementMultisensory integration is ubiquitous throughout the brain, including primary sensory cortices. The present study examined visual responses in primary auditory cortex, which were found in both putative excitatory and inhibitory neurons and concentrated in the deep cortical layers. Visual-modulated responses to sound were similarly observed in excitatory and inhibitory neurons but were more evenly distributed throughout cortical layers. Visual modulation moreover differed substantially across distinct sound response types. Transient stimulus onset spike rate changes were far less sensitive to visual context than sustained spike rate changes during the remainder of the stimulus. Spike timing changes were often modulated independently of spike rate changes. Audiovisual integration in auditory cortex is thus diversely expressed among cell types, cortical layers, and response types.
]]></description>
<dc:creator>Bigelow, J.</dc:creator>
<dc:creator>Morrill, R. J.</dc:creator>
<dc:creator>Olsen, T.</dc:creator>
<dc:creator>Bazarini, S. N.</dc:creator>
<dc:creator>Hasenstaub, A. R.</dc:creator>
<dc:date>2021-08-07</dc:date>
<dc:identifier>doi:10.1101/2021.08.06.455445</dc:identifier>
<dc:title><![CDATA[Visual modulation of spectrotemporal receptive fields in mouse auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.12.456045v1?rss=1">
<title>
<![CDATA[
Autophagy cargo receptors are secreted via extracellular vesicles and particles in response to endolysosomal inhibition or impaired autophagosome maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.12.456045v1?rss=1</link>
<description><![CDATA[
The endosome-lysosome (endolysosome) system plays central roles in both autophagic degradation and secretory pathways, including the exocytic release of extracellular vesicles and particles (EVPs). Although previous work has revealed important interconnections between autophagy and EVP-mediated secretion, our molecular understanding of these secretory events during endolysosome inhibition remains incomplete. Here, we delineate a secretory autophagy pathway upregulated in response to endolysosomal inhibition that mediates the EVP-associated extracellular release of autophagic cargo receptors, including p62/SQSTM1. This extracellular secretion is highly regulated and critically dependent on multiple ATGs required for the progressive steps of early autophagosome formation as well as Rab27a-dependent exocytosis. Furthermore, the disruption of autophagosome maturation, either due to genetic inhibition of the autophagosome-to-autolyosome fusion machinery or blockade via the SARS-CoV2 viral protein ORF3a, is sufficient to induce robust EVP-associated secretion of autophagy cargo receptors. Finally, we demonstrate that this ATG-dependent, EVP-mediated secretion pathway buffers against the intracellular accumulation of autophagy cargo receptors when classical autophagic degradation is impaired. Based on these results, we propose that secretory autophagy via EVPs functions as an alternate route to clear sequestered material and maintain proteostasis in response to endolysosomal dysfunction or impaired autophagosome maturation.
]]></description>
<dc:creator>Solvik, T. A.</dc:creator>
<dc:creator>Nguyen, T. A.</dc:creator>
<dc:creator>Lin, Y.-H. T.</dc:creator>
<dc:creator>Marsh, T.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:creator>Debnath, J.</dc:creator>
<dc:creator>Leidal, A. M.</dc:creator>
<dc:date>2021-08-12</dc:date>
<dc:identifier>doi:10.1101/2021.08.12.456045</dc:identifier>
<dc:title><![CDATA[Autophagy cargo receptors are secreted via extracellular vesicles and particles in response to endolysosomal inhibition or impaired autophagosome maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.15.456408v1?rss=1">
<title>
<![CDATA[
Respiratory Alkalosis Provokes Spike-Wave Discharges in Seizure- Prone Rats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.15.456408v1?rss=1</link>
<description><![CDATA[
Hyperventilation reliably provokes seizures in patients diagnosed with absence epilepsy. Despite this predictable patient response, the mechanisms that enable hyperventilation to powerfully activate absence seizure-generating circuits remain entirely unknown. Using the WAG/Rij rat, an established rodent model of absence epilepsy, we demonstrate that absence seizures are highly sensitive to arterial carbon dioxide, suggesting that seizure-generating circuits are sensitive to pH. Moreover, hyperventilation consistently activated neurons within the intralaminar nuclei of the thalamus, a structure implicated in seizure generation. We show that intralaminar thalamus also contains pH-sensitive neurons. Collectively, these observations suggest that hyperventilation activates pH-sensitive neurons of the intralaminar nuclei to provoke absence seizures.
]]></description>
<dc:creator>Salvati, K.</dc:creator>
<dc:creator>Souza, G. M. P. R.</dc:creator>
<dc:creator>Lu, A. C.</dc:creator>
<dc:creator>Ritger, M. L.</dc:creator>
<dc:creator>Guyenet, P.</dc:creator>
<dc:creator>Abbott, S. B.</dc:creator>
<dc:creator>Beenhakker, M. P.</dc:creator>
<dc:date>2021-08-15</dc:date>
<dc:identifier>doi:10.1101/2021.08.15.456408</dc:identifier>
<dc:title><![CDATA[Respiratory Alkalosis Provokes Spike-Wave Discharges in Seizure- Prone Rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.16.456540v1?rss=1">
<title>
<![CDATA[
Columnar localization and laminar origin of cortical surface electrical potentials 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.16.456540v1?rss=1</link>
<description><![CDATA[
Electrocorticography (ECoG) methodologically bridges basic neuroscience and understanding of human brains in health and disease. However, the localization of ECoG signals across the surface of the brain and the spatial distribution of their generating neuronal sources are poorly understood. To address this gap, we recorded from rat auditory cortex using customized ECoG, and simulated cortical surface electrical potentials with a full-scale, biophysically detailed cortical column model. Experimentally, ECoG-derived auditory representations were tonotopically organized and signals were anisotropically localized to [&le;]{+/-}200 m, i.e., a single cortical column. Biophysical simulations reproduce experimental findings, and indicate that neurons in cortical layers V and VI contribute [~]85% of evoked high-gamma signal recorded at the surface. Cell number and synchronicity were the primary biophysical properties determining laminar contributions to evoked ECoG signals, while distance was only a minimal factor. Thus, evoked ECoG signals primarily originate from neurons in the infragranular layers of a single cortical column.

In BriefBaratham et al., investigated the localization and origins of sensory evoked ECoG responses. They experimentally found that ECoG responses were anisotropically localized [&le;]{+/-}200 m, i.e., a single cortical column. Biophysically detailed simulations revealed that neurons in layers V &VI were the primary sources of evoked ECoG responses, in contrast to common thinking.

HighlightsEvoked ECoG signals are localized on the surface to a cortical column.

Neurons in cortical layers V and VI constitute the vast majority of the signal recorded at the surface.

Different laminar contributions to ECoG signal are driven by cell density and synchronicity.
]]></description>
<dc:creator>Baratham, V. L.</dc:creator>
<dc:creator>Dougherty, M. E.</dc:creator>
<dc:creator>Ledochowitsch, P.</dc:creator>
<dc:creator>Maharbiz, M. M.</dc:creator>
<dc:creator>Bouchard, K.</dc:creator>
<dc:date>2021-08-17</dc:date>
<dc:identifier>doi:10.1101/2021.08.16.456540</dc:identifier>
<dc:title><![CDATA[Columnar localization and laminar origin of cortical surface electrical potentials]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.16.456548v1?rss=1">
<title>
<![CDATA[
Precision engineering of an anti-HLA-A2 chimeric antigen receptor in regulatory T cells for transplant immune tolerance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.16.456548v1?rss=1</link>
<description><![CDATA[
Infusion of regulatory T cells (Tregs) engineered with a chimeric antigen receptor (CAR) targeting donor-derived human leukocyte antigen (HLA) is a promising strategy to promote transplant tolerance. Here, we describe an anti-HLA-A2 CAR (A2-CAR) generated by grafting the complementarity-determining regions (CDRs) of a human monoclonal anti-HLA-A2 antibody into the framework regions of the Herceptin 4D5 single-chain variable fragment and fusing it with a CD28-{zeta} signaling domain. The CDR-grafted A2-CAR maintained the specificity of the original antibody. We then generated HLA-A2 mono-specific human CAR Tregs either by deleting the endogenous T-cell receptor (TCR) via CRISPR/Cas9 and introducing the A2-CAR using lentiviral transduction or by directly integrating the CAR construct into the TCR alpha constant locus using homology-directed repair. These A2-CAR+TCRdeficient human Tregs maintained both Treg phenotype and function in vitro. Moreover, they selectively accumulated in HLA-A2-expressing islets transplanted from either HLA-A2 transgenic mice or deceased human donors. A2-CAR+TCRdeficient Tregs did not impair the function of these HLA-A2+ islets, whereas similarly engineered A2-CAR+TCRdeficientCD4+ conventional T cells rejected the islets in less than 2 weeks. A2-CAR+TCRdeficient Tregs delayed graft-versus-host disease only in the presence of HLA-A2, expressed either by co-transferred peripheral blood mononuclear cells or by the recipient mice. Altogether, we demonstrate that genome-engineered mono-antigen-specific A2-CAR Tregs localize to HLA-A2-expressing grafts and exhibit antigen-dependent in vivo suppression, independent of TCR expression. These approaches may be applied towards developing precision Treg cell therapies for transplant tolerance.
]]></description>
<dc:creator>Muller, Y. D.</dc:creator>
<dc:creator>Ferreira, L. M. R.</dc:creator>
<dc:creator>Ronin, E.</dc:creator>
<dc:creator>Ho, P.</dc:creator>
<dc:creator>Nguyen, V.</dc:creator>
<dc:creator>Faleo, G.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Lee, K.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Skartsis, N.</dc:creator>
<dc:creator>Kaul, A. M.</dc:creator>
<dc:creator>Mulder, A.</dc:creator>
<dc:creator>Claas, F. H. J.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:creator>Bluestone, J. A.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:date>2021-08-17</dc:date>
<dc:identifier>doi:10.1101/2021.08.16.456548</dc:identifier>
<dc:title><![CDATA[Precision engineering of an anti-HLA-A2 chimeric antigen receptor in regulatory T cells for transplant immune tolerance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.18.456686v1?rss=1">
<title>
<![CDATA[
Imaging-guided fine tuning of CAR synapse dynamics and T cell triggering 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.18.456686v1?rss=1</link>
<description><![CDATA[
T cells typically recognize their ligands using a defined cell biology - the scanning of their membrane microvilli to palpate their environment - while that same membrane scaffolds T cell receptors (TCRs) that can signal upon ligand binding. Chimeric antigen receptors (CARs) present both a therapeutic promise as well as a tractable means to study the interplay between receptor affinity, microvillar dynamics and T cell function. CARs are often built using single-chain variable fragments (scFvs) with far greater affinity than that of natural TCRs. We used high resolution lattice lightsheet (LLS) and total internal reflection fluorescence (TIRF) imaging to visualize microvillar scanning in the context of variations in CAR design. This demonstrated that conventional CARs hyper-stabilized microvillar contacts relative to TCRs. Reducing the affinity and/or avidity of binding brought synapse microvillar dynamics into natural ranges, normalized synapse resolution and improved downstream effector function. This work highlights the importance of understanding the underlying cell biology when designing receptors for optimal antigen engagement.
]]></description>
<dc:creator>Beppler, C.</dc:creator>
<dc:creator>Eichorst, J.</dc:creator>
<dc:creator>Marchuk, K.</dc:creator>
<dc:creator>Cai, E.</dc:creator>
<dc:creator>Castellanos, C. A.</dc:creator>
<dc:creator>Sriram, V.</dc:creator>
<dc:creator>Roybal, K. T.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2021-08-19</dc:date>
<dc:identifier>doi:10.1101/2021.08.18.456686</dc:identifier>
<dc:title><![CDATA[Imaging-guided fine tuning of CAR synapse dynamics and T cell triggering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.25.457626v1?rss=1">
<title>
<![CDATA[
The B.1.427/1.429 (epsilon) SARS-CoV-2 variants are more virulent than ancestral B.1 (614G) in Syrian hamsters 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.25.457626v1?rss=1</link>
<description><![CDATA[
As novel SARS-CoV-2 variants continue to emerge, it is critical that their potential to cause severe disease and evade vaccine-induced immunity is rapidly assessed in humans and studied in animal models. In early January 2021, a novel variant of concern (VOC) designated B.1.429 comprising 2 lineages, B.1.427 and B.1.429, was originally detected in California (CA) and shown to enhance infectivity in vitro and decrease antibody neutralization by plasma from convalescent patients and vaccine recipients. Here we examine the virulence, transmissibility, and susceptibility to pre-existing immunity for B 1.427 and B 1.429 in the Syrian hamster model. We find that both strains exhibit enhanced virulence as measured by increased body weight loss compared to hamsters infected with ancestral B.1 (614G), with B.1.429 causing the most body weight loss among all 3 lineages. Faster dissemination from airways to parenchyma and more severe lung pathology at both early and late stages were also observed with B.1.429 infections relative to B.1. (614G) and B.1.427 infections. In addition, subgenomic viral RNA (sgRNA) levels were highest in oral swabs of hamsters infected with B.1.429, however sgRNA levels in lungs were similar in all three strains. This demonstrates that B.1.429 replicates to higher levels than ancestral B.1 (614G) or B.1.427 in the upper respiratory tract (URT) but not in the lungs. In multi-virus in-vivo competition experiments, we found that epsilon (B.1.427/B.1.429) and gamma (P.1) dramatically outcompete alpha (B.1.1.7), beta (B.1.351) and zeta (P.2) in the lungs. In the URT gamma, and epsilon dominate, but the highly infectious alpha variant also maintains a moderate size niche. We did not observe significant differences in airborne transmission efficiency among the B.1.427, B.1.429 and ancestral B.1 (614G) variants in hamsters. These results demonstrate enhanced virulence and high relative fitness of the epsilon (B.1.427/B.1.429) variant in Syrian hamsters compared to an ancestral B.1 (614G) strain.

Author SummaryIn the last 12 months new variants of SARS-CoV-2 have arisen in the UK, South Africa, Brazil, India, and California. New SARS-CoV-2 variants will continue to emerge for the foreseeable future in the human population and the potential for these new variants to produce severe disease and evade vaccines needs to be understood. In this study, we used the hamster model to determine the epsilon (B.1.427/429) SARS-CoV-2 strains that emerged in California in late 2020 cause more severe disease and infected hamsters have higher viral loads in the upper respiratory tract compared to the prior B.1 (614G) strain. These findings are consistent with human clinical data and help explain the emergence and rapid spread of this strain in early 2021.
]]></description>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Fox, D.</dc:creator>
<dc:creator>van Doremalen, N.</dc:creator>
<dc:creator>Ball, E.</dc:creator>
<dc:creator>Morris, M. K.</dc:creator>
<dc:creator>Sotomayor-Gonzalez, A.</dc:creator>
<dc:creator>Servellita, V.</dc:creator>
<dc:creator>Rustagi, A.</dc:creator>
<dc:creator>Yinda, C. K.</dc:creator>
<dc:creator>Fritts, L.</dc:creator>
<dc:creator>Port, J. R.</dc:creator>
<dc:creator>Ma, Z.-M.</dc:creator>
<dc:creator>Holbrook, M.</dc:creator>
<dc:creator>Schulz, J.</dc:creator>
<dc:creator>Blish, C. A.</dc:creator>
<dc:creator>Hanson, C.</dc:creator>
<dc:creator>Chiu, C. Y.</dc:creator>
<dc:creator>Munster, V.</dc:creator>
<dc:creator>Stanley, S.</dc:creator>
<dc:creator>Miller, C. J.</dc:creator>
<dc:date>2021-08-25</dc:date>
<dc:identifier>doi:10.1101/2021.08.25.457626</dc:identifier>
<dc:title><![CDATA[The B.1.427/1.429 (epsilon) SARS-CoV-2 variants are more virulent than ancestral B.1 (614G) in Syrian hamsters]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.26.457842v1?rss=1">
<title>
<![CDATA[
IDENTIFICATION OF A NOVEL BDNF-SPECIFIC CORTICOSTRIATAL CIRCUITRY 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.26.457842v1?rss=1</link>
<description><![CDATA[
BDNF is released from axon terminals originating in the cerebral cortex onto striatal neurons. Here, we characterized BDNF neurons in the corticostriatal circuitry. First, we utilized BDNF-Cre and Ribotag transgenic mouse lines to label BDNF-positive neurons in the cortex and detected BDNF expression in all the subregions of the prefrontal cortex (PFC). Next, we used a retrograde viral tracing strategy, in combination with BDNF-Cre knockin mice, to map the cortical outputs of BDNF neurons in the dorsomedial and dorsolateral striatum (DMS and DLS). We found that BDNF-expressing neurons located in the medial prefrontal cortex (mPFC) project mainly to the DMS, and those located in the primary and secondary motor cortices (M1 and M2) and agranular insular cortex (AI) project mainly to the DLS. In contrast, BDNF-expressing orbitofrontal cortical (OFC) neurons differentially target the dorsal striatum (DS) depending on their mediolateral and rostrocaudal location. Specifically, the DMS is mainly innervated by the medial and ventral part of the orbitofrontal cortex (MO and VO) whereas the DLS receives projections specifically from the lateral part of the OFC (LO). Together, our study uncovers previously unknown BDNF corticostriatal circuitries. These findings could have important implications for the role of BDNF signaling in corticostriatal pathways.

Significance StatementBDNF is released in axons upon neuronal depolarization. Surprisingly, careful mapping of BDNF projecting neurons in the central nervous system (CNS) has not been conducted. Using retrograde viral strategies in combination with transgenic mice, we mapped out corticostriatal BDNF circuits. We found that, mPFC BDNF neurons project mainly to the DMS whereas the motor cortex and AI project to the DLS. BDNF neurons in the OFC are anatomically segregated. Whereas the DMS receives BDNF-positive projections from the VO, the DLS mainly receives BDNF-positive projections from the LO. Our findings could be important to the study of BDNF in corticostriatal circuitries.
]]></description>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>Soneja, D.</dc:creator>
<dc:creator>Phamluong, K.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2021-08-26</dc:date>
<dc:identifier>doi:10.1101/2021.08.26.457842</dc:identifier>
<dc:title><![CDATA[IDENTIFICATION OF A NOVEL BDNF-SPECIFIC CORTICOSTRIATAL CIRCUITRY]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.31.458413v1?rss=1">
<title>
<![CDATA[
Colloidal aggregators in biochemical SARS-CoV-2 repurposing screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.31.458413v1?rss=1</link>
<description><![CDATA[
To fight the SARS-CoV-2 pandemic, much effort has been directed toward drug repurposing, testing investigational and approved drugs against several viral or human proteins in vitro. Here we investigate the impact of colloidal aggregation, a common artifact in early drug discovery, in these repurposing screens. We selected 56 drugs reported to be active in biochemical assays and tested them for aggregation by both dynamic light scattering and by enzyme counter screening with and without detergent; seventeen of these drugs formed colloids at concentrations similar to their literature reported IC50s. To investigate the occurrence of colloidal aggregators more generally in repurposing libraries, we further selected 15 drugs that had physical properties resembling known aggregators from a common repurposing library, and found that 6 of these aggregated at micromolar concentrations. An attraction of repurposing is that drugs active on one target are considered de-risked on another. This study suggests not only that many of the drugs repurposed for SARS-CoV-2 in biochemical assays are artifacts, but that, more generally, when screened at relevant concentrations, drugs can act artifactually via colloidal aggregation. Understanding the role of aggregation, and detecting its effects rapidly, will allow the community to focus on those drugs and leads that genuinely have potential for treating COVID-19.



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org.highwire.dtl.DTLVardef@bfe5a2org.highwire.dtl.DTLVardef@7d05a1org.highwire.dtl.DTLVardef@1fcc3a5org.highwire.dtl.DTLVardef@190a153_HPS_FORMAT_FIGEXP  M_FIG Table of Contents Graphic

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]]></description>
<dc:creator>O'Donnell, H. R.</dc:creator>
<dc:creator>Tummino, T. A.</dc:creator>
<dc:creator>Bardine, C.</dc:creator>
<dc:creator>Craik, C. S.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2021-08-31</dc:date>
<dc:identifier>doi:10.1101/2021.08.31.458413</dc:identifier>
<dc:title><![CDATA[Colloidal aggregators in biochemical SARS-CoV-2 repurposing screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.02.458767v1?rss=1">
<title>
<![CDATA[
Mechanism of innate immune reprogramming by a fungal meningitis pathogen 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.02.458767v1?rss=1</link>
<description><![CDATA[
How deadly fungal pathogens overcome mammalian innate immunity is largely unknown. Cryptococcus neoformans, the most common cause of fungal meningitis, induces a pathogenic type 2 response characterized by pulmonary eosinophilia and alternatively activated macrophages. Using forward genetics, we identified a fungal secreted protein, Cpl1, necessary and sufficient to enhance alternative activation of primary macrophages in vitro. Cpl1-enhanced polarization requires Toll-like receptor 4, a known mediator of allergen-induced type 2 responses. Cpl1 is essential for virulence, drives polarization of interstitial macrophages in vivo, and requires type 2 cytokine signaling for its impact on infectivity. C. neoformans selectively associates with polarized interstitial macrophages during infection, supporting a direct hostpathogen interaction. This work identifies a secreted effector produced by a human fungal pathogen that reprograms innate immunity to enable tissue infection.

One sentence summaryIdentification of a secreted fungal effector that promotes virulence by enhancing type 2 inflammation
]]></description>
<dc:creator>Dang, E. V.</dc:creator>
<dc:creator>Lei, S.</dc:creator>
<dc:creator>Radkov, A.</dc:creator>
<dc:creator>Madhani, H.</dc:creator>
<dc:date>2021-09-02</dc:date>
<dc:identifier>doi:10.1101/2021.09.02.458767</dc:identifier>
<dc:title><![CDATA[Mechanism of innate immune reprogramming by a fungal meningitis pathogen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.02.458790v1?rss=1">
<title>
<![CDATA[
Biosynthesis of linear protein nanoarrays using the flagellar axoneme 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.02.458790v1?rss=1</link>
<description><![CDATA[
Applications in biotechnology and synthetic biology often make use of soluble proteins, but there are many potential advantages to anchoring enzymes to a stable substrate, including stability and the possibility for substrate channeling. To avoid the necessity of protein purification and chemical immobilization, there has been growing interest in bio-assembly of protein-containing nanoparticles, exploiting the self-assembly of viral capsid proteins or other proteins that form polyhedral structures. But these nanoparticle are limited in size which constrains the packaging and the accessibility of the proteins. The axoneme, the insoluble protein core of the eukaryotic flagellum or cilium, is a highly ordered protein structure that can be several microns in length, orders of magnitude larger than other types of nanoparticles. We show that when proteins of interest are fused to specific axonemal proteins and expressed in living cells, they become incorporated into linear arrays which have the advantages of high protein loading capacity, high stability, and single-step purification with retention of biomass. The arrays can be isolated as membrane enclosed vesicle or as exposed protein arrays. The approach is demonstrated for both fluorescent proteins and enzymes, and in the latter case it is found that incorporation into axoneme arrays provides increased stability for the enzyme.
]]></description>
<dc:creator>Ishikawa, H.</dc:creator>
<dc:creator>Tian, J. L.</dc:creator>
<dc:creator>Yu, J. E.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:creator>Qin, H.</dc:creator>
<dc:date>2021-09-02</dc:date>
<dc:identifier>doi:10.1101/2021.09.02.458790</dc:identifier>
<dc:title><![CDATA[Biosynthesis of linear protein nanoarrays using the flagellar axoneme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.02.458799v1?rss=1">
<title>
<![CDATA[
Hybrid ssDNA repair templates enable high yield genome engineering in primary cells for disease modeling and cell therapy manufacturing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.02.458799v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas9 offers unprecedented opportunities to modify genome sequences in primary human cells to study disease variants and reprogram cell functions for next-generation cellular therapies. CRISPR has several potential advantages over widely used retroviral vectors including: 1) site-specific transgene insertion via homology directed repair (HDR), and 2) reductions in the cost and complexity of genome modification. Despite rapid progress with ex vivo CRISPR genome engineering, many novel research and clinical applications would be enabled by methods to further improve knock-in efficiency and the absolute yield of live knock-in cells, especially with large HDR templates (HDRT). We recently reported that Cas9 target sequences (CTS) could be introduced into double-stranded DNA (dsDNA) HDRTs to improve knock-in, but yields and efficiencies were limited by toxicity at high HDRT concentrations. Here we developed a novel system that takes advantage of lower toxicity with single-stranded DNA (ssDNA). We designed hybrid ssDNA HDRTs that incorporate CTS sites and were able to boost knock-in percentages by >5-fold and live cell yields by >7-fold relative to dsDNA HDRTs with CTS. Knock-in efficiency and yield with ssCTS HDRTs were increased further with small molecule inhibitor combinations to improve HDR. We demonstrate application of these methods across a variety of target loci, knock-in constructs, and primary human cell types to reach ultra-high HDR efficiencies (>80-90%) which we use for pathogenic gene variant modeling and universal gene replacement strategies for IL2RA and CTLA4 mutations associated with mendelian immune disorders. Finally, we develop a GMP-compatible method for fully non-viral CAR-T cell manufacturing, demonstrating knock-in efficiencies of 46-62% and generating yields of >1.5 x 109 CAR+ T cells, well above current doses for adoptive cellular therapies. Taken together, we present a comprehensive non-viral approach to model disease associated mutations and re-write targeted genome sequences to program immune cell therapies at a scale compatible with future clinical application.
]]></description>
<dc:creator>Shy, B. R.</dc:creator>
<dc:creator>Vykunta, V.</dc:creator>
<dc:creator>Ha, A.</dc:creator>
<dc:creator>Roth, T. L.</dc:creator>
<dc:creator>Talbot, A.</dc:creator>
<dc:creator>Nguyen, D. N.</dc:creator>
<dc:creator>Chen, Y. Y.</dc:creator>
<dc:creator>Blaeschke, F.</dc:creator>
<dc:creator>Vedova, S.</dc:creator>
<dc:creator>Mamedov, M.</dc:creator>
<dc:creator>Chung, J.-Y.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Wolf, J.</dc:creator>
<dc:creator>Martin, T. G.</dc:creator>
<dc:creator>Ye, L.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Esensten, J.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2021-09-04</dc:date>
<dc:identifier>doi:10.1101/2021.09.02.458799</dc:identifier>
<dc:title><![CDATA[Hybrid ssDNA repair templates enable high yield genome engineering in primary cells for disease modeling and cell therapy manufacturing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.03.458835v1?rss=1">
<title>
<![CDATA[
Allosteric mechanism of signal transduction in the two-component system histidine kinase PhoQ 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.03.458835v1?rss=1</link>
<description><![CDATA[
Transmembrane signaling proteins couple extracytosolic sensors to cytosolic effectors. Here, we examine how binding of Mg2+ to the sensor domain of an E. coli two component histidine kinase (HK), PhoQ, modulates its cytoplasmic kinase domain. We use cysteine-crosslinking and reporter-gene assays to simultaneously and independently probe the signaling state of PhoQs sensor and autokinase domains in a set of over 30 mutants. Strikingly, conservative single-site mutants distant from the sensor or catalytic site strongly influence PhoQs ligand-sensitivity as well as the magnitude and direction of the signal, endowing diverse signaling characteristics without need for epistasis. Data from 35 mutants are explained by a semi-empirical 3-domain model in which the sensor, intervening HAMP, and catalytic domains can adopt kinase-promoting or inhibiting conformations, that are in allosteric communication. The catalytic and sensor domains intrinsically favor a constitutively  kinase-on conformation, while the HAMP favors the  off state; when coupled, they create a bistable system responsive to physiological [Mg2+]. Mutants alter signaling by locally modulating these intrinsic equilibrium constants and couplings. Our model suggests signals transmit via interdomain allostery rather than propagation of a single concerted conformational change, explaining the diversity of signaling structural transitions observed in individual HK domains.
]]></description>
<dc:creator>Mensa, B.</dc:creator>
<dc:creator>Polizzi, N. F.</dc:creator>
<dc:creator>Molnar, K. S.</dc:creator>
<dc:creator>Natale, A. M.</dc:creator>
<dc:creator>Lemmin, T.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:date>2021-09-05</dc:date>
<dc:identifier>doi:10.1101/2021.09.03.458835</dc:identifier>
<dc:title><![CDATA[Allosteric mechanism of signal transduction in the two-component system histidine kinase PhoQ]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.07.459343v1?rss=1">
<title>
<![CDATA[
A unique form of collective epithelial migration is crucial for tissue fusion in the secondary palate and can overcome loss of epithelial apoptosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.07.459343v1?rss=1</link>
<description><![CDATA[
Tissue fusion is an oft-employed process in morphogenesis which often requires the removal of the epithelia intervening multiple distinct primordia to form one continuous structure. In the mammalian secondary palate, a midline epithelial seam (MES) forms between two palatal shelves and must be removed to allow mesenchymal confluence. Abundant apoptosis and cell extrusion in this epithelial seam support their importance in its removal. However, by genetically disrupting the intrinsic apoptotic regulators BAX and BAK within the MES, we find a complete loss of cell death and cell extrusion, but successful removal of the MES, indicating that developmental compensation enables fusion. Novel static and live imaging approaches reveal that the MES is removed through a unique form of collective epithelial cell migration in which epithelial trails and islands stream through the mesenchyme to reach the oral and nasal epithelial surfaces. These epithelial trails and islands begin to express periderm markers while retaining expression of the basal epithelial marker {Delta}Np63, suggesting their migration to the oral and nasal surface is concomitant with their differentiation to an epithelial intermediate. Live imaging reveals anisotropic actomyosin contractility within epithelial trails that drives their peristaltic movement, and genetic loss of non-muscle myosin IIA-mediated actomyosin contractility results in dispersion of epithelial collectives and dramatic failure of normal MES migration. These findings demonstrate redundancy between cellular mechanisms of morphogenesis and reveal a crucial role for a unique form of collective epithelial migration during tissue fusion.
]]></description>
<dc:creator>Teng, T.</dc:creator>
<dc:creator>Teng, C.</dc:creator>
<dc:creator>Kaartinen, V.</dc:creator>
<dc:creator>Bush, J. O.</dc:creator>
<dc:date>2021-09-08</dc:date>
<dc:identifier>doi:10.1101/2021.09.07.459343</dc:identifier>
<dc:title><![CDATA[A unique form of collective epithelial migration is crucial for tissue fusion in the secondary palate and can overcome loss of epithelial apoptosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.08.459490v1?rss=1">
<title>
<![CDATA[
Spatio-temporal coordination at the maternal-fetal interface promotes trophoblast invasion and vascular remodeling in the first half of human pregnancy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.08.459490v1?rss=1</link>
<description><![CDATA[
Beginning in the first trimester, fetally derived extravillous trophoblasts (EVTs) invade the uterus and remodel its spiral arteries, transforming them into large, dilated blood vessels left with a thin, discontinuous smooth muscle layer and partially lined with EVTs. Several mechanisms have been proposed to explain how EVTs coordinate with the maternal decidua to promote a tissue microenvironment conducive to spiral artery remodeling (SAR). However, it remains a matter of debate which immune and stromal cell types participate in these interactions and how this process evolves with respect to gestational age. Here, we used a multiomic approach that combined the strengths of spatial proteomics and transcriptomics to construct the first spatiotemporal atlas of the human maternal-fetal interface in the first half of pregnancy. We used multiplexed ion beam imaging by time of flight (MIBI-TOF) and a 37-plex antibody panel to analyze [~]500,000 cells and 588 spiral arteries within intact decidua from 66 patients between 6-20 weeks of gestation, integrating this with coregistered transcriptomic profiles. Gestational age substantially influenced the frequency of many maternal immune and stromal cells, with tolerogenic subsets expressing CD206, CD163, TIM-3, Galectin-9, and IDO-1 increasingly enriched and colocalized at later time points. In contrast, SAR progression preferentially correlated with EVT invasion and was transcriptionally defined by 78 gene ontology pathways exhibiting unique monotonic and biphasic trends. Lastly, we developed an integrated model of SAR supporting an intravasation mechanism where invasion is accompanied by upregulation of pro-angiogenic, immunoregulatory EVT programs that promote interactions with vascular endothelium while avoiding activation of immune cells in circulating maternal blood. Taken together, these results support a coordinated model of decidualization in which increasing gestational age drives a transition in maternal decidua towards a tolerogenic niche conducive to locally regulated, EVT-dependent SAR.
]]></description>
<dc:creator>Greenbaum, S.</dc:creator>
<dc:creator>Averbukh, I.</dc:creator>
<dc:creator>Soon, E.</dc:creator>
<dc:creator>Rizzuto, G.</dc:creator>
<dc:creator>Baranski, A.</dc:creator>
<dc:creator>Greenwald, N.</dc:creator>
<dc:creator>Bosse, M.</dc:creator>
<dc:creator>Jaswa, E. G.</dc:creator>
<dc:creator>Khair, Z.</dc:creator>
<dc:creator>Kwok, S.</dc:creator>
<dc:creator>Warshawsky, S.</dc:creator>
<dc:creator>Miller, G.</dc:creator>
<dc:creator>Schwartz, M.</dc:creator>
<dc:creator>Graf, W.</dc:creator>
<dc:creator>Van Valen, D.</dc:creator>
<dc:creator>Keren, L.</dc:creator>
<dc:creator>Hollmann, T.</dc:creator>
<dc:creator>van de Rijn, M.</dc:creator>
<dc:creator>Angelo, M.</dc:creator>
<dc:date>2021-09-10</dc:date>
<dc:identifier>doi:10.1101/2021.09.08.459490</dc:identifier>
<dc:title><![CDATA[Spatio-temporal coordination at the maternal-fetal interface promotes trophoblast invasion and vascular remodeling in the first half of human pregnancy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.10.457001v1?rss=1">
<title>
<![CDATA[
Stimulation related artifacts and a multipurpose template-based offline removal solution for a novel sensing-enabled deep brain stimulation device 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.10.457001v1?rss=1</link>
<description><![CDATA[
BackgroundThe Medtronic "Percept" is the first FDA approved deep brain stimulation (DBS) device with sensing capabilities during active stimulation. Its real-world signal recording properties have yet to be fully described.

ObjectiveThis study details sources of artifact (and potential mitigations) in local field potential (LFP) signals collected by the Percept, and assesses the potential impact of artifact on the future development of adaptive DBS (aDBS) using this device.

MethodsLFP signals were collected from seven subjects in both experimental and clinical settings. The presence of artifacts and their effect on the spectral content of neural signals were evaluated in both the stimulation ON and OFF states using three distinct offline artifact removal techniques.

ResultsTemplate subtraction successfully removed multiple sources of artifact, including 1) electrocardiogram (ECG), 2) non-physiologic polyphasic artifacts, and 3) ramping related artifacts seen when changing stimulation amplitudes. ECG removal from stimulation ON (at 0 mA) signals recovered the spectral shape seen when OFF stimulation (averaged difference in normalized power in theta, alpha, and beta bands [&le;] 3.5%). ECG removal using singular value decomposition was similarly successful, though required subjective researcher input. QRS interpolation produced similar recovery of beta-band signal, but resulted in residual low-frequency artifact.

ConclusionsArtifacts present when stimulation is enabled notably affected the spectral properties of sensed signals using the Percept. Multiple discrete artifacts could be successfully removed offline using an automated template subtraction method. The presence of unrejected artifact likely influences online power estimates, with the potential to affect aDBS algorithm performance.
]]></description>
<dc:creator>Hammer, L. H.</dc:creator>
<dc:creator>Kochanski, R. B.</dc:creator>
<dc:creator>Starr, P.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:date>2021-09-11</dc:date>
<dc:identifier>doi:10.1101/2021.09.10.457001</dc:identifier>
<dc:title><![CDATA[Stimulation related artifacts and a multipurpose template-based offline removal solution for a novel sensing-enabled deep brain stimulation device]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.15.460398v1?rss=1">
<title>
<![CDATA[
Integrated gene analyses of de novo mutations from 46,612 trios with autism and developmental disorders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.15.460398v1?rss=1</link>
<description><![CDATA[
Most genetic studies consider autism spectrum disorder (ASD) and developmental disorder (DD) separately despite overwhelming comorbidity and shared genetic etiology. Here we analyzed de novo mutations (DNMs) from 15,560 ASD (6,557 are new) and 31,052 DD trios independently and combined as broader neurodevelopmental disorders (NDD) using three models. We identify 615 candidate genes (FDR 5%, 189 potentially novel) by one or more models, including 138 reaching exome-wide significance (p < 3.64e-07) in all models. We find no evidence for ASD-specific genes in contrast to 18 genes significantly enriched for DD. There are 53 genes show particular mutational-bias including enrichments for missense (n=41) or truncating DNM (n=12). We find 22 genes with evidence of sex-bias including five X chromosome genes also with significant female burden (DDX3X, MECP2, SMC1A, WDR45, and HDAC8). NDD risk genes group into five functional networks associating with different brain developmental lineages based on single-cell nuclei transcriptomic data, which provides important insights into disease subtypes and future functional studies.
]]></description>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Kim, C.</dc:creator>
<dc:creator>Bakken, T. E.</dc:creator>
<dc:creator>Gillentine, M. A.</dc:creator>
<dc:creator>Henning, B.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Gilissen, C.</dc:creator>
<dc:creator>Consortium, S.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:date>2021-09-16</dc:date>
<dc:identifier>doi:10.1101/2021.09.15.460398</dc:identifier>
<dc:title><![CDATA[Integrated gene analyses of de novo mutations from 46,612 trios with autism and developmental disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.15.460552v1?rss=1">
<title>
<![CDATA[
Divergent encoding of active avoidance behavior in corticostriatal and corticolimbic projections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.15.460552v1?rss=1</link>
<description><![CDATA[
Active avoidance behavior, in which an animal performs an action to avoid a stressor, is crucial for survival and may provide insight into avoidance behaviors seen in anxiety disorders. Active avoidance requires the dorsomedial prefrontal cortex (dmPFC), which is thought to regulate avoidance via downstream projections to the striatum and amygdala. However, the endogenous activity of projection-defined dmPFC subpopulations during active avoidance learning remains unexplored. Here we utilized fiber photometry to record from the dmPFC and its downstream projections to the dorsomedial striatum (DMS) and the basolateral amygdala (BLA) during active avoidance learning in mice. We examined neural activity during conditioned stimulus (CS) presentations, active avoidance, and cued freezing. Both prefrontal projections showed learning-related increases in activity during CS onset throughout active avoidance training. The dmPFC as a whole showed increased activity during avoidance and decreased activity during cued freezing. Finally, dmPFC-DMS and dmPFC-BLA projections showed divergent encoding of active avoidance behavior, with the dmPFC-DMS projection showing increased activity and the dmPFC-BLA showing decreased activity during active avoidance. Our results identify differential prefrontal encoding of active and passive coping behaviors in the same behavioral paradigm and demonstrate divergent encoding of active avoidance in projection-specific dmPFC subpopulations.
]]></description>
<dc:creator>Kajs, B. L.</dc:creator>
<dc:creator>Loewke, A. C.</dc:creator>
<dc:creator>Dorsch, J. M.</dc:creator>
<dc:creator>Vinson, L. T.</dc:creator>
<dc:creator>Gunaydin, L. A.</dc:creator>
<dc:date>2021-09-16</dc:date>
<dc:identifier>doi:10.1101/2021.09.15.460552</dc:identifier>
<dc:title><![CDATA[Divergent encoding of active avoidance behavior in corticostriatal and corticolimbic projections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.20.460546v1?rss=1">
<title>
<![CDATA[
Genome-wide mapping implicates R-loops in lineage-specific processes and formation of DNA break hotspots in neural stem/progenitor cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.20.460546v1?rss=1</link>
<description><![CDATA[
Recent work has revealed classes of recurrent DNA double-strand breaks (DSBs) in neural stem/progenitor cells, including transcription-associated, promoter-proximal breaks and recurrent DSB clusters in late-replicating, long neural genes. However, the mechanistic factors promoting these different classes of DSBs in neural stem/progenitor cells are not understood. Here, we elucidated the genome-wide landscape of DNA:RNA hybrid structures called "R-loops" in primary neural stem/progenitor cells in order to assess their contribution to the different classes of DNA break "hotspots". We report that R-loops in neural stem/progenitor cells are associated primarily with transcribed regions that replicate early and genes that show GC skew in their promoter region. Surprisingly, the majority of genes with recurrent DSB clusters in long, neural genes does not show substantial R-loop accumulation. We implicate R-loops in promoter-proximal DNA break formation in highly transcribed, early replicating regions and conclude that R-loops are not a driver of recurrent double-strand break cluster formation in most long, neural genes. Together, our study provides an understanding of how R-loops may contribute to DNA break hotspots and affect lineage-specific processes in neural stem/progenitor cells.
]]></description>
<dc:creator>Thongthip, S.</dc:creator>
<dc:creator>Carlson, A.</dc:creator>
<dc:creator>Crossley, M. P.</dc:creator>
<dc:creator>Schwer, B.</dc:creator>
<dc:date>2021-09-20</dc:date>
<dc:identifier>doi:10.1101/2021.09.20.460546</dc:identifier>
<dc:title><![CDATA[Genome-wide mapping implicates R-loops in lineage-specific processes and formation of DNA break hotspots in neural stem/progenitor cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.21.461269v1?rss=1">
<title>
<![CDATA[
Ligand binding remodels protein side chain conformational heterogeneity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.21.461269v1?rss=1</link>
<description><![CDATA[
While protein conformational heterogeneity plays an important role in many aspects of biological function, including ligand binding, its impact has been difficult to quantify. Macromolecular X-ray diffraction is commonly interpreted with a static structure, but it can provide information on both the anharmonic and harmonic contributions to conformational heterogeneity. Here, through multiconformer modeling of time- and space-averaged electron density, we measure conformational heterogeneity of 743 stringently matched pairs of crystallographic datasets that reflect unbound/apo and ligand-bound/holo states. When comparing the conformational heterogeneity of side chains, we observe that when binding site residues become more rigid upon ligand binding, distant residues tend to become more flexible, especially in non-solvent exposed regions. Among ligand properties, we observe increased protein flexibility as the number of hydrogen bonds decrease and relative hydrophobicity increases. Across a series of 13 inhibitor bound structures of CDK2, we find that conformational heterogeneity is correlated with inhibitor features and identify how conformational changes propagate differences in conformational heterogeneity away from the binding site. Collectively, our findings agree with models emerging from NMR studies suggesting that residual side chain entropy can modulate affinity and point to the need to integrate both static conformational changes and conformational heterogeneity in models of ligand binding.
]]></description>
<dc:creator>Wankowicz, S. A.</dc:creator>
<dc:creator>de Oliveira, S. H. P.</dc:creator>
<dc:creator>Hogan, D. W.</dc:creator>
<dc:creator>van den Bedem, H.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2021-09-21</dc:date>
<dc:identifier>doi:10.1101/2021.09.21.461269</dc:identifier>
<dc:title><![CDATA[Ligand binding remodels protein side chain conformational heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.21.461284v1?rss=1">
<title>
<![CDATA[
The nuclear transport factor CSE1 drives macronuclear volume increase and macronuclear node condensation in Stentor coeruleus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.21.461284v1?rss=1</link>
<description><![CDATA[
The giant ciliate, Stentor coeruleus, provides a unique opportunity to study nuclear shape because its macronucleus undergoes a rapid, dramatic, and developmentally regulated shape change. During a 2 hour time period within cell division and regeneration, the 400 um long moniliform macronucleus coalesces into a single mass, elongates into a vermiform shape, and then renodulates, returning to its original beads-on-a-string morphology.1 Previous work from the 1960s - 1980s demonstrated that the macronuclear shape change is a highly regulated part of cell division and regeneration 2,3, but the molecular pathways driving these changes are unknown. With the recent availability of a sequenced Stentor genome, a transcriptome during regeneration, and molecular tools like RNAi 4-6, it is now possible to investigate the molecular mechanisms that drive macronuclear shape change. We found that the volume of the macronucleus increases during coalescence, suggesting an inflation-based mechanism. When the nuclear transport factor, CSE1, is knocked down by RNAi, the shape and volume changes of the macronucleus are attenuated, and nuclear morphology is altered. CSE1 protein undergoes a dynamic relocalization correlated with nuclear shape changes, being mainly cytoplasmic prior to nuclear coalescence, and accumulating inside the macronucleus during coalescence. At the end of regeneration, CSE1 is degraded during the period when the macronucleus returns to its pre-coalescence volume. We propose a model in which nuclear transport via CSE1 increases the volume of the macronucleus, thereby decreasing the surface to volume ratio and driving coalescence of the many nodes into a single mass.
]]></description>
<dc:creator>McGillivary, R. M.</dc:creator>
<dc:creator>Sood, P.</dc:creator>
<dc:creator>Hammar, K.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2021-09-22</dc:date>
<dc:identifier>doi:10.1101/2021.09.21.461284</dc:identifier>
<dc:title><![CDATA[The nuclear transport factor CSE1 drives macronuclear volume increase and macronuclear node condensation in Stentor coeruleus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.21.461298v1?rss=1">
<title>
<![CDATA[
Perilacunar bone tissue exhibits sub-micrometer modulus gradation which depends on the recency of osteocyte bone formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.21.461298v1?rss=1</link>
<description><![CDATA[
Osteocytes are capable of resorbing and replacing bone local to the lacunar-canalicular system (LCS remodeling). However, the impacts of these processes on perilacunar bone quality are not understood. It is well established that aging is associated with reduced whole-bone fracture resistance, reduced osteocyte viability, and truncated LCS geometries, but it remains unclear if aging changes perilacunar bone quality. In this study, we employed atomic force microscopy (AFM) to quantify sub-micrometer gradations from 2D maps surrounding osteocyte lacunae in young (5 mo) and aged (22 mo) female mice. AFM-mapped lacunae were also imaged with confocal laser scanning microscopy to determine which osteocytes had recently deposited bone as determined by the presence of fluorochrome labels. These assays allowed us to quantify gradations in nanoscale mechanical properties of bone-forming/non-bone-forming osteocytes in young and aged mice. This study reports for the first time that there are sub-micrometer gradations in modulus surrounding lacunae and that these gradations are dependent upon recent osteocyte bone formation. Perilacunar bone adjacent to bone-forming osteocytes demonstrated lower peak and bulk modulus values when compared to bone near non-bone-forming osteocytes from the same mouse. Bone-forming osteocytes also showed increased perilacunar modulus variability. Age reduced lacunar size but did not significant effect modulus gradation or variability. In general, lacunar morphology was not a strong predictor of modulus gradation patterns. These findings support the idea that lacunar-canalicular remodeling activity changes the material properties of surrounding bone tissue on a sub-micrometer scale. Therefore, conditions that affect osteocyte health have the potential to impact bone quality.

GRAPHICAL ABSTRACT

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]]></description>
<dc:creator>Rux, C. J.</dc:creator>
<dc:creator>Vahidi, G.</dc:creator>
<dc:creator>Darabi, A.</dc:creator>
<dc:creator>Cox, L. M.</dc:creator>
<dc:creator>Heveran, C. M.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.21.461298</dc:identifier>
<dc:title><![CDATA[Perilacunar bone tissue exhibits sub-micrometer modulus gradation which depends on the recency of osteocyte bone formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.23.461579v1?rss=1">
<title>
<![CDATA[
Multi-context genetic modeling of transcriptional regulation resolves novel disease loci 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.23.461579v1?rss=1</link>
<description><![CDATA[
A majority of the variants identified in genome-wide association studies fall in non-coding regions of the genome, indicating their mechanism of impact is mediated via gene expression. Leveraging this hypothesis, transcriptome-wide association studies (TWAS) have assisted in both the interpretation and discovery of additional genes associated with complex traits. However, existing methods for conducting TWAS do not take full advantage of the intra-individual correlation inherently present in multi-context expression studies and do not properly adjust for multiple testing across contexts. We developed CONTENT-- a computationally efficient method with proper cross-context false discovery correction that leverages correlation structure across contexts to improve power and generate context-specific and context-shared components of expression. We applied CONTENT to bulk multi-tissue and single-cell RNA-seq data sets and show that CONTENT leads to a 42% (bulk) and 110% (single cell) increase in the number of genetically predicted genes relative to previous approaches. Interestingly, we find the context-specific component of expression comprises 30% of heritability in tissue-level bulk data and 75% in single-cell data, consistent with cell type heterogeneity in bulk tissue. In the context of TWAS, CONTENT increased the number of gene-phenotype associations discovered by over 47% relative to previous methods across 22 complex traits.
]]></description>
<dc:creator>Thompson, M.</dc:creator>
<dc:creator>Gordon, M. G.</dc:creator>
<dc:creator>Lu, A.</dc:creator>
<dc:creator>Tandon, A.</dc:creator>
<dc:creator>Halperin, E.</dc:creator>
<dc:creator>Gusev, A.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Balliu, B.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:date>2021-09-23</dc:date>
<dc:identifier>doi:10.1101/2021.09.23.461579</dc:identifier>
<dc:title><![CDATA[Multi-context genetic modeling of transcriptional regulation resolves novel disease loci]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.24.461597v1?rss=1">
<title>
<![CDATA[
Polygenic enrichment distinguishes disease associations of individual cells in single-cell RNA-seq data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.24.461597v1?rss=1</link>
<description><![CDATA[
Gene expression at the individual cell-level resolution, as quantified by single-cell RNA-sequencing (scRNA-seq), can provide unique insights into the pathology and cellular origin of diseases and complex traits. Here, we introduce single-cell Disease Relevance Score (scDRS), an approach that links scRNA-seq with polygenic risk of disease at individual cell resolution without the need for annotation of individual cells to cell types; scDRS identifies individual cells that show excess expression levels for genes in a disease-specific gene set constructed from GWAS data. We determined via simulations that scDRS is well-calibrated and powerful in identifying individual cells associated to disease. We applied scDRS to GWAS data from 74 diseases and complex traits (average N =346K) in conjunction with 16 scRNA-seq data sets spanning 1.3 million cells from 31 tissues and organs. At the cell type level, scDRS broadly recapitulated known links between classical cell types and disease, and also produced novel biologically plausible findings. At the individual cell level, scDRS identified subpopulations of disease-associated cells that are not captured by existing cell type labels, including subpopulations of CD4+ T cells associated with inflammatory bowel disease, partially characterized by their effector-like states; subpopulations of hippocampal CA1 pyramidal neurons associated with schizophrenia, partially characterized by their spatial location at the proximal part of the hippocampal CA1 region; and subpopulations of hepatocytes associated with triglyceride levels, partially characterized by their higher ploidy levels. At the gene level, we determined that genes whose expression across individual cells was correlated with the scDRS score (thus reflecting co-expression with GWAS disease genes) were strongly enriched for gold-standard drug target and Mendelian disease genes.
]]></description>
<dc:creator>Zhang, M. J.</dc:creator>
<dc:creator>Hou, K.</dc:creator>
<dc:creator>Dey, K. K.</dc:creator>
<dc:creator>Jagadeesh, K. A.</dc:creator>
<dc:creator>Weinand, K.</dc:creator>
<dc:creator>Sakaue, S.</dc:creator>
<dc:creator>Taychameekiatchai, A.</dc:creator>
<dc:creator>Rao, P.</dc:creator>
<dc:creator>Pisco, A. O.</dc:creator>
<dc:creator>Zou, J.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Gandal, M.</dc:creator>
<dc:creator>Raychaudhuri, S.</dc:creator>
<dc:creator>Pasaniuc, B.</dc:creator>
<dc:creator>Price, A. L.</dc:creator>
<dc:date>2021-09-28</dc:date>
<dc:identifier>doi:10.1101/2021.09.24.461597</dc:identifier>
<dc:title><![CDATA[Polygenic enrichment distinguishes disease associations of individual cells in single-cell RNA-seq data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.25.461797v1?rss=1">
<title>
<![CDATA[
Single-molecule analysis of specificity and multivalency in binding of short linear substrate motifs to the APC/C 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.25.461797v1?rss=1</link>
<description><![CDATA[
Robust regulatory signals in the cell often depend on interactions between short linear motifs (SLiMs) and globular proteins. Many of these interactions are poorly characterized because the binding proteins cannot be produced in the amounts needed for traditional methods. To address this problem, we developed a single-molecule off-rate (SMOR) assay based on microscopy of fluorescent ligand binding to immobilized protein partners. We used it to characterize substrate binding to the Anaphase-Promoting Complex/Cyclosome (APC/C), a ubiquitin ligase that triggers chromosome segregation. We find that SLiMs in APC/C substrates (the D box and KEN box) display distinct affinities and specificities for the substrate-binding subunits of the APC/C, and we show that multiple SLiMs in a substrate generate a high-affinity multivalent interaction. The remarkably adaptable substrate-binding mechanisms of the APC/C have the potential to govern the order of substrate destruction in mitosis.
]]></description>
<dc:creator>Hartooni, N.</dc:creator>
<dc:creator>Sung, J.</dc:creator>
<dc:creator>Jain, A.</dc:creator>
<dc:creator>Morgan, D. O.</dc:creator>
<dc:date>2021-09-26</dc:date>
<dc:identifier>doi:10.1101/2021.09.25.461797</dc:identifier>
<dc:title><![CDATA[Single-molecule analysis of specificity and multivalency in binding of short linear substrate motifs to the APC/C]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.27.462016v1?rss=1">
<title>
<![CDATA[
The short flagella 1 (SHF1) gene in Chlamydomonas encodes a Crescerin TOG-domain protein required for late stages of flagellar growth. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.27.462016v1?rss=1</link>
<description><![CDATA[
Length control of flagella represents a simple and tractable system to investigate the dynamics of organelle size. Models for flagellar length control in the model organism, Chlamydomonas reinhardtii have focused on the length-dependence of the intraflagellar transport (IFT) system which manages the delivery and removal of axonemal subunits at the tip of the flagella. One of these cargoes, tubulin, is the major axonemal subunit, and its frequency of arrival at the tip plays a central role in size control models. However, the mechanisms determining tubulin dynamics at the tip are still poorly understood. We discovered a loss-of-function mutation that leads to shortened flagella, and found that this was an allele of a previously described gene, SHF1, whose molecular identity had not previously been determined. We found that SHF1 encodes a Chlamydomonas ortholog of Crescerin, previously identified as a cilia-specific TOG-domain array protein that can bind tubulin via its TOG domains and increase tubulin polymerization rates. In this mutant, flagellar regeneration occurs with the same initial kinetics as wild-type cells, but plateaus at a shorter length. Using a computational model in which the flagellar microtubules are represented by a differential equation for flagellar length combined with a stochastic model for cytoplasmic microtubule dynamics, we found that our experimental results are best described by a model in which Crescerin/SHF1 binds tubulin dimers in the cytoplasm and transports them into the flagellum. We suggest that this TOG-domain protein is necessary to efficiently and preemptively increase intra-flagella tubulin levels to offset decreasing IFT cargo at the tip as flagellar assembly progresses.
]]></description>
<dc:creator>Perlaza, K.</dc:creator>
<dc:creator>Mirvis, M.</dc:creator>
<dc:creator>Ishikawa, H.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2021-09-27</dc:date>
<dc:identifier>doi:10.1101/2021.09.27.462016</dc:identifier>
<dc:title><![CDATA[The short flagella 1 (SHF1) gene in Chlamydomonas encodes a Crescerin TOG-domain protein required for late stages of flagellar growth.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.29.462463v1?rss=1">
<title>
<![CDATA[
Optogenetic EB1 inactivation shortens metaphase spindles by disrupting cortical force-producing interactions with astral microtubules 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.29.462463v1?rss=1</link>
<description><![CDATA[
Chromosome segregation is accomplished by the mitotic spindle, a bipolar micromachine built primarily from microtubules. Different microtubule populations contribute to spindle function: Kinetochore microtubules attach and transmit forces to chromosomes, antiparallel interpolar microtubules support spindle structure, and astral microtubules connect spindle poles to the cell cortex [1,2]. In mammalian cells, End Binding (EB) proteins associate with all growing microtubule plus ends throughout the cell cycle and serve as adaptors for a diverse group of +TIPs that control microtubule dynamics and interactions with other intracellular structures [3]. Because binding of many +TIPs to EB1 and thus microtubule-end association is switched off by mitotic phosphorylation [4-6] the mitotic function of EBs remains poorly understood. To analyze how EB1 and associated +TIPs on different spindle microtubule populations contribute to mitotic spindle dynamics, we use a light sensitive EB1 variant, {pi}-EB1, that allows local, acute and reversible inactivation of +TIP association with growing microtubule ends in live cells [7]. We find that acute {pi}-EB1 photoinactivation results in rapid and reversible metaphase spindle shortening and transient relaxation of tension across the central spindle. However, in contrast to interphase, {pi}-EB1 photoinactivation does not inhibit microtubule growth in metaphase, but instead increases astral microtubule length and number. Yet, in the absence of EB1 activity astral microtubules fail to engage the cortical dynein/dynactin machinery and spindle poles move away from regions of {pi}-EB1 photoinactivation. In conclusion, our optogenetic approach reveals mitotic EB1 functions that remain hidden in genetic experiments likely due to compensatory molecular systems regulating vertebrate spindle dynamics.
]]></description>
<dc:creator>Dema, A.</dc:creator>
<dc:creator>van Haren, J.</dc:creator>
<dc:creator>Wittmann, T.</dc:creator>
<dc:date>2021-10-01</dc:date>
<dc:identifier>doi:10.1101/2021.09.29.462463</dc:identifier>
<dc:title><![CDATA[Optogenetic EB1 inactivation shortens metaphase spindles by disrupting cortical force-producing interactions with astral microtubules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.29.462487v1?rss=1">
<title>
<![CDATA[
Endoplasmic reticulum stress activates human IRE1α through reversible assembly of inactive dimers into small oligomers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.29.462487v1?rss=1</link>
<description><![CDATA[
Protein folding homeostasis in the endoplasmic reticulum (ER) is regulated by a signaling network, termed the unfolded protein response (UPR). Inositol-requiring enzyme 1 (IRE1) is an ER membrane-resident kinase/RNase that mediates signal transmission in the most evolutionarily conserved branch of the UPR. Dimerization and/or higher-order oligomerization of IRE1 are thought to be important for its activation mechanism, yet the actual oligomeric states of inactive, active, and attenuated mammalian IRE1 complexes remained unknown. We developed an automated two-color single-molecule tracking approach to dissect the oligomerization of tagged endogenous human IRE1 in live cells. In contrast to previous models, our data indicate that IRE1 exists as a constitutive homodimer at baseline and assembles into small oligomers upon ER stress. We demonstrate that the formation of inactive dimers and stress-dependent oligomers is fully governed by IRE1s lumenal domain. Phosphorylation of IRE1s kinase domain occurs more slowly than oligomerization and is retained after oligomers disassemble back into dimers. Our findings suggest that assembly of IRE1 dimers into larger oligomers specifically enables trans- autophosphorylation, which in turn drives IRE1s RNase activity.
]]></description>
<dc:creator>Belyy, V.</dc:creator>
<dc:creator>Zuazo-Gaztelu, I.</dc:creator>
<dc:creator>Alamban, A.</dc:creator>
<dc:creator>Ashkenazi, A.</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:date>2021-09-30</dc:date>
<dc:identifier>doi:10.1101/2021.09.29.462487</dc:identifier>
<dc:title><![CDATA[Endoplasmic reticulum stress activates human IRE1α through reversible assembly of inactive dimers into small oligomers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.01.462825v1?rss=1">
<title>
<![CDATA[
Structural basis of ATP-dependent high-fidelity epigenome maintenance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.01.462825v1?rss=1</link>
<description><![CDATA[
Epigenetic evolution occurs over million-year timescales in Cryptococcus neoformans and is mediated by DNMT5, the first maintenance-type cytosine methyltransferase identified in the fungal or protist kingdoms. DNMT5 requires ATP and displays exquisite hemimethyl-DNA specificity. To understand these novel properties, we solved cryo-EM structures of CnDNMT5 in three states. These studies reveal an elaborate allosteric cascade in which hemimethylated DNA first activates the SNF2 ATPase domain by a large rigid body rotation while the target cytosine partially flips out the DNA duplex. ATP binding then triggers a striking structural reconfiguration of the methyltransferase catalytic pocket that enables cofactor binding, completion of base-flipping, and catalysis. Unmethylated DNA binding fails to open cofactor pocket and subsequent ATP binding triggers its ejection to ensure fidelity. This chaperone-like, enzyme-remodeling role of the SNF2 domain illuminates how energy can be used to enable faithful epigenetic memory.

HighlightsO_LIStructures of DNMT5 reveal mechanism of ATP-dependent DNA methylation
C_LIO_LIHemimethylated CpG recognition triggers partial base flipping of the target cytosine
C_LIO_LIHemimethylated DNA induces rigid body rotation to activate the SNF2 ATPase domain
C_LIO_LIMTase catalytic pocket is remodeled by the SNF2 ATPase to achieve specificity
C_LI
]]></description>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Catania, S.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>de la Cruz, M. J.</dc:creator>
<dc:creator>Rao, B.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:creator>Patel, D. J.</dc:creator>
<dc:date>2021-10-01</dc:date>
<dc:identifier>doi:10.1101/2021.10.01.462825</dc:identifier>
<dc:title><![CDATA[Structural basis of ATP-dependent high-fidelity epigenome maintenance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.04.463005v1?rss=1">
<title>
<![CDATA[
Robust T cell activation requires an eIF3-driven burst in T cell receptor translation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.04.463005v1?rss=1</link>
<description><![CDATA[
Activation of T cells requires a rapid surge in cellular protein synthesis. However, the role of translation initiation in the early induction of specific genes remains unclear. Here we show human translation initiation factor eIF3 interacts with select immune system related mRNAs including those encoding the T cell receptor (TCR) subunits TCRA and TCRB. Binding of eIF3 to the TCRA and TCRB mRNA 3-untranslated regions (3-UTRs) depends on CD28 coreceptor signaling and regulates a burst in TCR translation required for robust T cell activation. Use of the TCRA or TCRB 3-UTRs to control expression of an anti-CD19 chimeric antigen receptor (CAR) improves the ability of CAR-T cells to kill tumor cells in vitro. These results identify a new mechanism of eIF3-mediated translation control that can aid T cell engineering for immunotherapy applications.
]]></description>
<dc:creator>DeSilva, D.</dc:creator>
<dc:creator>Ferguson, L.</dc:creator>
<dc:creator>Chin, G.</dc:creator>
<dc:creator>Smith, B.</dc:creator>
<dc:creator>Apathy, R.</dc:creator>
<dc:creator>Roth, T.</dc:creator>
<dc:creator>Blaeschke, F.</dc:creator>
<dc:creator>Kudla, M.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Ingolia, N. T.</dc:creator>
<dc:creator>Cate, J. H.</dc:creator>
<dc:date>2021-10-04</dc:date>
<dc:identifier>doi:10.1101/2021.10.04.463005</dc:identifier>
<dc:title><![CDATA[Robust T cell activation requires an eIF3-driven burst in T cell receptor translation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.05.462047v1?rss=1">
<title>
<![CDATA[
Pan-cancer imaging of TERT expression using deuterium magnetic resonance spectroscopy-based assessment of pyruvate metabolism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.05.462047v1?rss=1</link>
<description><![CDATA[
Telomerase reverse transcriptase (TERT) expression is indispensable for tumor immortality. Non-invasive methods of imaging TERT can, therefore, report on tumor proliferation and response to therapy. Here, we show that TERT expression is associated with elevated levels of the redox metabolite NADH in multiple cancers, including glioblastoma, oligodendroglioma, melanoma, neuroblastoma, and hepatocellular carcinoma. Mechanistically, TERT acts via the metabolic regulator FOXO1 to upregulate nicotinamide phosphoribosyl transferase, which is the key enzyme for NADH biosynthesis. Importantly, deuterium magnetic resonance spectroscopy (2H-MRS), which is a novel, clinically translatable metabolic imaging modality, can be leveraged for imaging TERT-linked NADH in preclinical tumor models in vivo. Since NADH is essential for pyruvate flux to lactate, 2H-MRS following administration of 2H-labeled pyruvate non-invasively visualizes TERT expression and reports on early response to therapy. Collectively, our study provides insights into the mechanisms of TERT-linked metabolic reprogramming and, importantly, establishes 2H-MRS as a pan-cancer strategy for imaging tumor immortality.
]]></description>
<dc:creator>Batsios, G.</dc:creator>
<dc:creator>Taglang, C.</dc:creator>
<dc:creator>Tran, M.</dc:creator>
<dc:creator>Stevers, N.</dc:creator>
<dc:creator>Barger, C.</dc:creator>
<dc:creator>Gillespie, A. M.</dc:creator>
<dc:creator>Ronen, S.</dc:creator>
<dc:creator>Costello, J.</dc:creator>
<dc:creator>Viswanath, P.</dc:creator>
<dc:date>2021-10-06</dc:date>
<dc:identifier>doi:10.1101/2021.10.05.462047</dc:identifier>
<dc:title><![CDATA[Pan-cancer imaging of TERT expression using deuterium magnetic resonance spectroscopy-based assessment of pyruvate metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.06.463386v1?rss=1">
<title>
<![CDATA[
Aberrant Functional Connectivity between Reward and Inhibitory Control Networks in Pre-Adolescent Binge Eating Disorder 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.06.463386v1?rss=1</link>
<description><![CDATA[
BackgroundBehavioral features of binge eating disorder (BED) suggest abnormalities in reward and inhibitory control. Studies of adult populations suggest functional abnormalities in reward and inhibitory control networks. Despite behavioral markers often developing in children, the neurobiology of pediatric BED remains unstudied.

Methods58 pre-adolescent children (aged 9-10-years) with BED and 66 age, BMI and developmentally-matched control children were extracted from the 3.0 baseline (Year 0) release of the Adolescent Brain Cognitive Development (ABCD) Study. We investigated group differences in resting-state functional MRI (rs-fMRI) functional connectivity (FC) within and between reward and inhibitory control networks. A seed-based approach was employed to assess nodes in the reward (orbitofrontal cortex, nucleus accumbens, amygdala) and inhibitory control (dorsolateral prefrontal cortex, anterior cingulate cortex) networks via hypothesis-driven seed-to- seed analyses, and secondary seed-to-voxel analyses.

ResultsOur findings revealed reduced FC between the dlPFC and amygdala, and between the anterior cingulate cortex and orbitofrontal cortex in pre-adolescent children with BED, relative to age, gender, BMI and developmentally matched controls. These findings indicating aberrant connectivity between nodes of inhibitory control and reward networks were corroborated by the whole-brain FC analyses.

ConclusionsEarly-onset BED may be characterized by diffuse abnormalities in the functional synergy between reward and cognitive control networks, without perturbations within reward and inhibitory control networks, respectively. The decreased capacity to regulate a reward-driven pursuit of hedonic foods, which is characteristic of BED, may in part, rest on this dysconnectivity between reward and inhibitory control networks.
]]></description>
<dc:creator>Murray, S.</dc:creator>
<dc:creator>Alba, C.</dc:creator>
<dc:creator>Duval, C.</dc:creator>
<dc:creator>Nagata, J.</dc:creator>
<dc:creator>Cabeen, R.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Toga, A. W.</dc:creator>
<dc:creator>Siegel, S.</dc:creator>
<dc:creator>Jann, K.</dc:creator>
<dc:date>2021-10-08</dc:date>
<dc:identifier>doi:10.1101/2021.10.06.463386</dc:identifier>
<dc:title><![CDATA[Aberrant Functional Connectivity between Reward and Inhibitory Control Networks in Pre-Adolescent Binge Eating Disorder]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.08.463652v1?rss=1">
<title>
<![CDATA[
A sense-antisense RNA interaction promotes breast cancer metastasis via regulation of NQO1 expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.08.463652v1?rss=1</link>
<description><![CDATA[
Antisense RNAs are ubiquitous in human cells, yet the role that they play in healthy and diseased states remains largely unexplored. Here, we developed a computational framework to catalog and profile antisense RNAs and applied it to poorly and highly metastatic breast cancer cell lines. We identified one antisense RNA that plays a functional role in driving breast cancer progression by upregulating the redox enzyme NQO1, and hence named NQO1-antisense RNA or NQO1-AS. This upregulation occurs via a stabilizing interaction between NQO1-AS and its complementary region in the 3UTR of NQO1 mRNA. By increasing expression of NQO1 protein, breast cancer cells are able to tolerate higher levels of oxidative stress, enabling them to colonize the lung. During this process the cancer cells become dependent on NQO1 to protect them from ferroptosis. We have shown that this dependence can be exploited therapeutically in xenograft models of metastasis. Together, our findings establish a previously unknown role for NQO1-AS in the progression of breast cancer by serving as a post-transcriptional regulator of RNA processing and decay for its sense mRNA.
]]></description>
<dc:creator>Culbertson, B.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Markett, D.</dc:creator>
<dc:creator>Asgharian, H.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Fish, L.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Woo, B.</dc:creator>
<dc:creator>Nanda, S.</dc:creator>
<dc:creator>Rabinowitz, J.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2021-10-09</dc:date>
<dc:identifier>doi:10.1101/2021.10.08.463652</dc:identifier>
<dc:title><![CDATA[A sense-antisense RNA interaction promotes breast cancer metastasis via regulation of NQO1 expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.12.464152v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 spike protein induces abnormal inflammatory blood clots neutralized by fibrin immunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.12.464152v1?rss=1</link>
<description><![CDATA[
Blood clots are a central feature of coronavirus disease-2019 (COVID-19) and can culminate in pulmonary embolism, stroke, and sudden death. However, it is not known how abnormal blood clots form in COVID-19 or why they occur even in asymptomatic and convalescent patients. Here we report that the Spike protein from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) binds to the blood coagulation factor fibrinogen and induces structurally abnormal blood clots with heightened proinflammatory activity. SARS-CoV-2 Spike virions enhanced fibrin-mediated microglia activation and induced fibrinogen-dependent lung pathology. COVID-19 patients had fibrin autoantibodies that persisted long after acute infection. Monoclonal antibody 5B8, targeting the cryptic inflammatory fibrin epitope, inhibited thromboinflammation. Our results reveal a procoagulant role for the SARS-CoV-2 Spike and propose fibrin-targeting interventions as a treatment for thromboinflammation in COVID-19.

One-Sentence SummarySARS-CoV-2 spike induces structurally abnormal blood clots and thromboinflammation neutralized by a fibrin-targeting antibody.
]]></description>
<dc:creator>Ryu, J. K.</dc:creator>
<dc:creator>Sozmen, E. G.</dc:creator>
<dc:creator>Dixit, K.</dc:creator>
<dc:creator>Montano, M.</dc:creator>
<dc:creator>Matsui, Y.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Helmy, E.</dc:creator>
<dc:creator>Deerinck, T. J.</dc:creator>
<dc:creator>Yan, Z.</dc:creator>
<dc:creator>Schuck, R.</dc:creator>
<dc:creator>Acevedo, R. M.</dc:creator>
<dc:creator>Spencer, C. M.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Pico, A. R.</dc:creator>
<dc:creator>Zamvil, S. S.</dc:creator>
<dc:creator>Lynch, K. L.</dc:creator>
<dc:creator>Ellisman, M. H.</dc:creator>
<dc:creator>Greene, W. C.</dc:creator>
<dc:creator>Akassoglou, K.</dc:creator>
<dc:date>2021-10-13</dc:date>
<dc:identifier>doi:10.1101/2021.10.12.464152</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 spike protein induces abnormal inflammatory blood clots neutralized by fibrin immunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.15.464597v1?rss=1">
<title>
<![CDATA[
Mapping enzyme catalysis with metabolomic biosensing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.15.464597v1?rss=1</link>
<description><![CDATA[
Enzymes are represented across a vast space of protein sequences and structural forms and have activities that far exceed the best chemical catalysts; however, engineering them to have novel or enhanced activity is limited by technologies for sensing product formation. Here, we describe a general and scalable approach for characterizing enzyme activity that uses the metabolism of the host cell as a biosensor by which to infer product formation. Since different products consume different molecules in their synthesis, they perturb host metabolism in unique ways that can be measured by mass spectrometry. This provides a general way by which to sense product formation, to discover unexpected products and map the effects of mutagenesis.
]]></description>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Chang, K.-C.</dc:creator>
<dc:creator>Payne, E. M.</dc:creator>
<dc:creator>Modavi, C.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Palmer, C.</dc:creator>
<dc:creator>Tao, N.</dc:creator>
<dc:creator>Alper, H. S.</dc:creator>
<dc:creator>Kennedy, R. T.</dc:creator>
<dc:creator>Cornett, D. S.</dc:creator>
<dc:creator>Abate, A. R.</dc:creator>
<dc:date>2021-10-16</dc:date>
<dc:identifier>doi:10.1101/2021.10.15.464597</dc:identifier>
<dc:title><![CDATA[Mapping enzyme catalysis with metabolomic biosensing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.17.464699v1?rss=1">
<title>
<![CDATA[
The ESX-1 secretion system senses bacterial contact and prepares mycobacteria for environmental adaptation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.17.464699v1?rss=1</link>
<description><![CDATA[
The ESX-1 system (6-kDa early secretory antigenic target (ESAT-6) secretion system-1) is essential for Mycobacterium tuberculosis pathogenesis and conjugal transfer in Mycobacterium smegmatis, yet little is known about how its function is regulated. Live-cell fluorescence microscopy showed natively expressed ESX-1 was organized into distinct foci predominantly observed at cell-cell contacts. These foci formed when two cells touched and required a fully assembled ESX-1 system in both bacteria, suggesting the generation of an ESX-1 megacomplex across multiple membranes. The emergence of ESX-1 foci and ESX-1 secretion was environmentally dependent: foci formed in low nitrogen environments in which secretion was suppressed, yet with increasing concentrations of nitrogen, ESX-1 systems diffused along the plasma membrane and secretion was activated. Genome-wide transcriptional profiling revealed ESX-1 dependent induction of genes required for the SOS response and error prone DNA replication in high nitrogen. Based on these findings, we propose a new model of ESX-1 function where ESX-1 localization and secretion are responsive to nitrogen levels and form an integral node in the mycobacterial response to neighboring cells and environmental adaptation.
]]></description>
<dc:creator>Herrera, N.</dc:creator>
<dc:creator>Odermatt, P. D.</dc:creator>
<dc:creator>Voorhies, M.</dc:creator>
<dc:creator>Nakagawa, R.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:creator>Chang, F.</dc:creator>
<dc:creator>Rosenberg, O. S.</dc:creator>
<dc:date>2021-10-17</dc:date>
<dc:identifier>doi:10.1101/2021.10.17.464699</dc:identifier>
<dc:title><![CDATA[The ESX-1 secretion system senses bacterial contact and prepares mycobacteria for environmental adaptation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.24.465635v1?rss=1">
<title>
<![CDATA[
Multi-model Order ICA: A Data-driven Method for Evaluating Brain Functional Network Connectivity Within and Between Multiple Spatial Scales 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.24.465635v1?rss=1</link>
<description><![CDATA[
BackgroundWhile functional connectivity is widely studied, there has been little work studying functional connectivity at different spatial scales. Likewise, the relationship of functional connectivity between spatial scales is unknown.

MethodsWe proposed an independent component analysis (ICA) - based approach to capture information at multiple model orders (component numbers) and to evaluate functional network connectivity (FNC) both within and between model orders. We evaluated the approach by studying group differences in the context of a study of resting fMRI (rsfMRI) data collected from schizophrenia (SZ) individuals and healthy controls (HC). The predictive ability of FNC at multiple spatial scales was assessed using support vector machine (SVM)-based classification.

ResultsIn addition to consistent predictive patterns at both multiple-model orders and single model orders, unique predictive information was seen at multiple-model orders and in the interaction between model orders. We observed that the FNC between model order 25 and 50 maintained the highest predictive information between HC and SZ. Results highlighted the predictive ability of the somatomotor and visual domains both within and between model orders compared to other functional domains. Also, subcortical-somatomotor, temporal-somatomotor, and temporal-subcortical FNCs had relatively high weights in predicting SZ.

ConclusionsIn sum, multi-model order ICA provides a more comprehensive way to study FNC, produces meaningful and interesting results which are applicable to future studies. We shared the spatial templates from this work at different model orders to provide a reference for the community, which can be leveraged in regression-based or fully automated (spatially constrained) ICA approaches.

Impact StatementMulti-model order ICA provides a comprehensive way to study brain functional network connectivity within and between multiple spatial scales, highlighting findings that would have been ignored in single model order analysis. This work expands upon and adds to the relatively new literature on resting fMRI-based classification and prediction. Results highlighted the differentiating power of specific intrinsic connectivity networks on classifying brain disorders of schizophrenia patients and healthy participants, at different spatial scales. The spatial templates from this work provide a reference for the community, which can be leveraged in regression-based or fully automated ICA approaches.
]]></description>
<dc:creator>Meng, X.</dc:creator>
<dc:creator>Iraji, A.</dc:creator>
<dc:creator>Fu, Z.</dc:creator>
<dc:creator>Kochunov, P.</dc:creator>
<dc:creator>Belger, A.</dc:creator>
<dc:creator>Ford, J.</dc:creator>
<dc:creator>McEwen, S.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>Mueller, B. A.</dc:creator>
<dc:creator>Pearlson, G. D.</dc:creator>
<dc:creator>Potkin, S. G.</dc:creator>
<dc:creator>Preda, A.</dc:creator>
<dc:creator>Turner, J.</dc:creator>
<dc:creator>van Erp, T. G. M.</dc:creator>
<dc:creator>Sui, J.</dc:creator>
<dc:creator>Calhoun, V.</dc:creator>
<dc:date>2021-10-27</dc:date>
<dc:identifier>doi:10.1101/2021.10.24.465635</dc:identifier>
<dc:title><![CDATA[Multi-model Order ICA: A Data-driven Method for Evaluating Brain Functional Network Connectivity Within and Between Multiple Spatial Scales]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.25.461994v1?rss=1">
<title>
<![CDATA[
Aldehyde dehydrogenase 3A1 deficiency leads to mitochondrial dysfunction and impacts salivary gland stem cell self-renewal, differentiation and survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.25.461994v1?rss=1</link>
<description><![CDATA[
Adult salivary stem/progenitor cells (SSPC) have an intrinsic property to self-renew in order to maintain tissue architecture and homeostasis. Adult salivary glands have been documented to harbor SSPC, which have been shown to play a vital role in the regeneration of the glandular structures post radiation damage. We have previously demonstrated that activation of aldehyde dehydrogenase 3A1 (ALDH3A1) after radiation reduced aldehyde accumulation in SSPC, leading to less apoptosis and improved salivary function. We subsequently found that sustained pharmacological ALDH3A1 activation is critical to enhance regeneration of murine submandibular gland after radiation damage. Further investigation shows that ALDH3A1 function is crucial for SSPC self-renewal and differentiation even in the absence of radiation stress. Salivary glands from Aldh3a1-null mice have fewer acinar structures than wildtype mice. ALDH3A1 deletion or pharmacological inhibition in SSPC leads to a decrease in mitochondrial DNA copy number, lower expression of mitochondrial specific genes and proteins, structural abnormalities, lower membrane potential, and reduced cellular respiration. Loss or inhibition of ALDH3A1 also elevates ROS levels and accumulation of ALDH3A1 substrate 4-hydroxynonenal (4-HNE, a lipid peroxidation product), leading to decreased survival of murine SSPC that can be rescued by treatment with 4-HNE specific carbonyl scavengers. Our data indicate that ALDH3A1 activity protects mitochondrial function and is important for the development and regeneration activity of SSPC. This knowledge will help to guide our translational strategy of applying ALDH3A1 activators in the clinic to prevent radiation-related hyposalivation in head and neck cancer patients.
]]></description>
<dc:creator>Viswanathan, V.</dc:creator>
<dc:creator>Cao, H.</dc:creator>
<dc:creator>Saiki, J.</dc:creator>
<dc:creator>Jiang, D.</dc:creator>
<dc:creator>Mattingly, A.</dc:creator>
<dc:creator>Nambiar, D. K.</dc:creator>
<dc:creator>Bloomstein, J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Jiang, S.</dc:creator>
<dc:creator>Chamoli, M.</dc:creator>
<dc:creator>Sirjani, D.</dc:creator>
<dc:creator>Kaplan, M.</dc:creator>
<dc:creator>Holsinger, F. C.</dc:creator>
<dc:creator>Liang, R.</dc:creator>
<dc:creator>Feng, Z.</dc:creator>
<dc:creator>Eyben, R. V.</dc:creator>
<dc:creator>Lagory, E.</dc:creator>
<dc:creator>Guan, L.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Ye, J.</dc:creator>
<dc:creator>Nolan, G. P.</dc:creator>
<dc:creator>Denko, N.</dc:creator>
<dc:creator>Knox, S.</dc:creator>
<dc:creator>Rosen, D. M.</dc:creator>
<dc:creator>Le, Q. T.</dc:creator>
<dc:date>2021-10-26</dc:date>
<dc:identifier>doi:10.1101/2021.10.25.461994</dc:identifier>
<dc:title><![CDATA[Aldehyde dehydrogenase 3A1 deficiency leads to mitochondrial dysfunction and impacts salivary gland stem cell self-renewal, differentiation and survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.27.466175v1?rss=1">
<title>
<![CDATA[
Deep Phenotypic Analysis of Blood and Lymphoid T and NK Cells from HIV+ Controllers and Non-Controllers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.27.466175v1?rss=1</link>
<description><![CDATA[
T and natural killer (NK) cells are effector cells with key roles in anti-HIV immunity, including in lymphoid tissues, the major site of HIV persistence. In this study, we used 42-parameter CyTOF to conduct deep phenotyping of paired blood- and lymph node (LN)-derived T and NK cells from three groups of HIV+ aviremic individuals: elite controllers, and antiretroviral therapy (ART)-suppressed individuals who had started therapy during chronic vs. acute infection, the latter of which is associated with better outcomes. We found that acute-treated individuals are enriched for specific subsets of T and NK cells, including blood-derived CD56-CD16+ NK cells previously associated with HIV control, and LN-derived CD4+ T follicular helper cells with heightened expansion potential. An in-depth comparison of the features of the cells from blood vs. LNs of individuals from our cohort revealed that T cells from blood were more activated than those from LNs. By contrast, LNs were enriched for follicle-homing CXCR5+ CD8+ T cells, which expressed increased levels of inhibitory receptors and markers of survival and proliferation as compared to their CXCR5-counterparts. In addition, a subset of memory-like CD56brightTCF1+ NK cells was enriched in LNs relative to blood. These results together suggest unique T and NK cell features in acute-treated individuals, and highlight the importance of examining effector cells not only in blood but also the lymphoid tissue compartment, where the reservoir mostly persists, and where these cells take on distinct phenotypic features.
]]></description>
<dc:creator>George, A. F.</dc:creator>
<dc:creator>Luo, X.</dc:creator>
<dc:creator>Neidleman, J.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Vohra, P.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Shin, M.-G.</dc:creator>
<dc:creator>Lee, M. J.</dc:creator>
<dc:creator>Blish, C. A.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Greene, W. C.</dc:creator>
<dc:creator>Lee, S. A.</dc:creator>
<dc:creator>Roan, N. R.</dc:creator>
<dc:date>2021-10-28</dc:date>
<dc:identifier>doi:10.1101/2021.10.27.466175</dc:identifier>
<dc:title><![CDATA[Deep Phenotypic Analysis of Blood and Lymphoid T and NK Cells from HIV+ Controllers and Non-Controllers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.31.466660v1?rss=1">
<title>
<![CDATA[
Chlamydomonas ARMC2/PF27 is an obligate cargo adapter for IFT of radial spokes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.31.466660v1?rss=1</link>
<description><![CDATA[
Intraflagellar transport (IFT) carries proteins into flagella but how IFT trains interact with the large number of diverse proteins required to assemble flagella remains largely unknown. Here, we show that IFT of radial spokes in Chlamydomonas requires ARMC2/PF27, a conserved armadillo repeat protein associated with male infertility and reduced lung function. Chlamydomonas ARMC2 was highly enriched in growing flagella and tagged ARMC2 and the spoke protein RSP3 comigrated on anterograde trains. In contrast, a cargo and an adapter of inner and outer dynein arms moved independently of ARMC2, indicating that unrelated cargoes distribute stochastically onto the IFT trains. After concomitant unloading at the flagellar tip, RSP3 attached to the axoneme whereas ARMC2 diffused back to the cell body. In armc2/pf27 mutants, IFT of radial spokes was abolished and the presence of radial spokes was limited to the proximal region of flagella. We conclude that ARMC2 is a cargo adapter required for IFT of radial spokes to ensure their assembly along flagella. ARMC2 belongs to a growing class of cargo-specific adapters that enable flagellar transport of preassembled axonemal substructures by IFT.
]]></description>
<dc:creator>Lechtreck, K. F.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Dai, J.</dc:creator>
<dc:creator>Alkhofash, R.</dc:creator>
<dc:creator>Butler, J.</dc:creator>
<dc:creator>Alford, L.</dc:creator>
<dc:creator>Yang, P.</dc:creator>
<dc:date>2021-10-31</dc:date>
<dc:identifier>doi:10.1101/2021.10.31.466660</dc:identifier>
<dc:title><![CDATA[Chlamydomonas ARMC2/PF27 is an obligate cargo adapter for IFT of radial spokes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.31.466670v1?rss=1">
<title>
<![CDATA[
Host Variation in Interferon, MHC Class I, Glycosylation, and Viral Transcription Genes Predict HIV Persistence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.31.466670v1?rss=1</link>
<description><![CDATA[
ObjectivePrior genomewide association studies have identified variation in MHC Class I alleles and CCR5{Delta}32 as genetic predictors of viral control, especially in "elite" controllers, individuals who remain virally suppressed in the absence of therapy.

DesignCross-sectional genomewide association study.

MethodsWe analyzed custom whole exome sequencing and direct HLA typing from 202 ART-suppressed HIV+ non-controllers in relation to four measures of the peripheral CD4+ T cell reservoir: HIV intact DNA, total (t)DNA, unspliced (us)RNA, and RNA/DNA. Linear mixed models were adjusted for potential covariates including age, sex, nadir CD4+ T cell count, pre-ART HIV RNA, timing of ART initiation, and duration of ART suppression.

ResultsPreviously reported "protective" host genetic mutations related to viral setpoint (e.g., among elite controllers) were found to predict smaller HIV reservoir size. The HLA "protective" B*57:01 was associated with significantly lower HIV usRNA (q=3.3x10-3), and among the largest subgroup, European ancestry individuals, the CCR5{Delta}32 deletion was associated with smaller HIV tDNA (p=4.3x10-3) and usRNA (p=8.7x10-3). In addition, genomewide analysis identified several SNPs in MX1 (an interferon stimulated gene) that were significantly associated with HIV tDNA (q=0.02), and the direction of these associations paralleled MX1 gene eQTL expression.

ConclusionsWe observed a significant association between previously reported "protective" MHC class I alleles and CCR5{Delta}32 with the HIV reservoir size in non-controllers. We also found a novel association between MX1 and HIV total DNA (in addition to other interferon signaling relevant genes, PPP1CB, DDX3X). These findings warrant further investigation in future validation studies.
]]></description>
<dc:creator>Siegel, D. A.</dc:creator>
<dc:creator>Thanh, C.</dc:creator>
<dc:creator>Wan, E.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Hobbs, K.</dc:creator>
<dc:creator>Pan, T.</dc:creator>
<dc:creator>Gibson, E. A.</dc:creator>
<dc:creator>Kroetz, D. L.</dc:creator>
<dc:creator>Hunt, P. W.</dc:creator>
<dc:creator>Marin, J.</dc:creator>
<dc:creator>Hecht, F.</dc:creator>
<dc:creator>Pilcher, C.</dc:creator>
<dc:creator>Milush, J.</dc:creator>
<dc:creator>Martin, M.</dc:creator>
<dc:creator>Carrington, M.</dc:creator>
<dc:creator>Pillai, S.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Huang, M.-L.</dc:creator>
<dc:creator>Roychoundhury, P.</dc:creator>
<dc:creator>Haworth, K.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Kiem, H.-P.</dc:creator>
<dc:creator>Heinrich, T. J.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:date>2021-11-03</dc:date>
<dc:identifier>doi:10.1101/2021.10.31.466670</dc:identifier>
<dc:title><![CDATA[Host Variation in Interferon, MHC Class I, Glycosylation, and Viral Transcription Genes Predict HIV Persistence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.05.467513v1?rss=1">
<title>
<![CDATA[
Single-Cell Multiomics Defines Tolerogenic Extrathymic Aire-Expressing Populations with Unique Homology to Thymic Epithelium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.05.467513v1?rss=1</link>
<description><![CDATA[
The Autoimmune Regulator (Aire) gene, well defined for its role in medullary thymic epithelial cells (mTECs) and immune self-tolerance, is also expressed in extrathymic Aire-expressing cells (eTACs) in the secondary lymphoid organs. eTACs have been shown to be hematopoietic antigen presenting cells (APCs) and potent inducers of immune tolerance (1-3). However, the precise identity and function of these cells remain unclear. Here, we use high-dimensional single-cell multiomics and functional approaches to define eTACs at the transcriptional, genomic, and proteomic level. We find that eTACs consist of two similar cell types: CCR7+ Aire-expressing migratory dendritic cells (AmDCs) and a unique Aire-hi population co-expressing Aire and RAR-related orphan receptor gamma-t (ROR{gamma}t). The latter, which have significant transcriptional and genomic homology to migratory dendritic cells (migDCs) and mTECs, we term Janus cells (JCs). All eTACs, and JCs in particular, have a highly accessible chromatin structure and high levels of broad gene expression, including tissue-specific antigens, as well as remarkable transcriptional and genomic homology to thymic medullary epithelium. As in the thymus, Aire expression in eTACs is also dependent on RANK-RANK-ligand interactions. Furthermore, lineage-tracing shows that JCs are not precursors to the majority of AmDCs. Finally, self-antigen expression by eTACs is sufficient to mediate negative selection of T cells escaping thymic selection and can prevent autoimmune diabetes in non-obese diabetic mice. This transcriptional, genomic, and functional symmetry between a hematopoietic Aire-expressing population in the periphery and an epithelial Aire-expressing population in the thymus suggests that a core biological program may influence self-tolerance and self-representation across the spectrum of immune development.
]]></description>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Lareau, C. A.</dc:creator>
<dc:creator>Bautista, J. L.</dc:creator>
<dc:creator>Gupta, A. R.</dc:creator>
<dc:creator>Sandor, K.</dc:creator>
<dc:creator>Germino, J.</dc:creator>
<dc:creator>Yin, Y.</dc:creator>
<dc:creator>Arvedson, M. P.</dc:creator>
<dc:creator>Reeder, G. C.</dc:creator>
<dc:creator>Cramer, N. T.</dc:creator>
<dc:creator>Xie, F.</dc:creator>
<dc:creator>Ntranos, V.</dc:creator>
<dc:creator>Satpathy, A. T.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:date>2021-11-06</dc:date>
<dc:identifier>doi:10.1101/2021.11.05.467513</dc:identifier>
<dc:title><![CDATA[Single-Cell Multiomics Defines Tolerogenic Extrathymic Aire-Expressing Populations with Unique Homology to Thymic Epithelium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.08.467750v1?rss=1">
<title>
<![CDATA[
CRISPR-Cas9 mediated nuclear transport and genomic integration of nanostructured genes in human primary cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.08.467750v1?rss=1</link>
<description><![CDATA[
DNA nanostructures are a promising tool for delivery of a variety of molecular payloads to cells. DNA origami structures, where 1000s of bases are folded into a compact nanostructure, present an attractive approach to package genes; however, effective delivery of genetic material into cell nuclei has remained a critical challenge. Here we describe the use of DNA nanostructures encoding an intact human gene and a fluorescent-protein encoding gene as compact templates for gene integration by CRISPR-mediated homology-directed repair (HDR). Our design includes CRISPR-Cas9 ribonucleoprotein (RNP) binding sites on the DNA nanostructures to increase shuttling of structures into the nucleus. We demonstrate efficient shuttling and genomic integration of DNA nanostructures using transfection and electroporation. These nanostructured templates display lower toxicity and higher insertion efficiency compared to unstructured double-stranded DNA (dsDNA) templates in human primary cells. Furthermore, our study validates virus-like particles (VLPs) as an efficient method of DNA nanostructure delivery, opening the possibility of delivering DNA nanostructures in vivo to specific cell types. Together these results provide new approaches to gene delivery with DNA nanostructures and establish their use as large HDR templates, exploiting both their design features and their ability to encode genetic information. This work also opens a door to translate other DNA nanodevice functions, such as measuring biophysical properties, into cell nuclei.

Teaser SentenceCRISPR-Cas9 mediates nuclear transport and integration of nanostructured genes in human primary cells
]]></description>
<dc:creator>Lin Shiao, E.</dc:creator>
<dc:creator>Pfeifer, W. G.</dc:creator>
<dc:creator>Shy, B. R.</dc:creator>
<dc:creator>Doost, M. S.</dc:creator>
<dc:creator>Chen, E.</dc:creator>
<dc:creator>Vykunta, V. S.</dc:creator>
<dc:creator>Hamilton, J. R.</dc:creator>
<dc:creator>Stahl, E. C.</dc:creator>
<dc:creator>Lopez, D. M.</dc:creator>
<dc:creator>Espinoza, C. R. S.</dc:creator>
<dc:creator>Dejanov, A. E.</dc:creator>
<dc:creator>Lew, R. J.</dc:creator>
<dc:creator>Poirer, M. G.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Castro, C. E.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:date>2021-11-09</dc:date>
<dc:identifier>doi:10.1101/2021.11.08.467750</dc:identifier>
<dc:title><![CDATA[CRISPR-Cas9 mediated nuclear transport and genomic integration of nanostructured genes in human primary cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.08.467777v1?rss=1">
<title>
<![CDATA[
Prevalent mouse phenotypes in the unexplored druggable genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.08.467777v1?rss=1</link>
<description><![CDATA[
Among the estimated ~23,000 protein encoding human genes, the class of  druggable genes- defined by their ability to bind drug-like compounds- represents an enticing collection of targets for clinical intervention. Yet many if not most of these genes remain poorly understood and understudied. Here we evaluate three major classes of druggable genes (GPCRs, ion channels, and kinases) and found that a third of these remain largely ignored yet display significant mouse phenotypes upon genetic ablation. We show that both well-studied and understudied druggable genes share a similar number and spectrum of phenotypes. Moreover, many of the mouse phenotypes arising from the ablation of both well-studied and understudied druggable genes show similarities with symptoms in rare human diseases. Collectively these data diminish the notion that most poorly studied genes may not be especially  important and highlight therapeutic opportunities and potential disease models among poorly characterized druggable genes.
]]></description>
<dc:creator>Gulyaeva, O.</dc:creator>
<dc:creator>Hu, Z.</dc:creator>
<dc:creator>Oprea, T. I.</dc:creator>
<dc:creator>Lloyd, K.</dc:creator>
<dc:creator>Gomez, S. M.</dc:creator>
<dc:creator>Roth, B. L.</dc:creator>
<dc:creator>McManus, M.</dc:creator>
<dc:date>2021-11-08</dc:date>
<dc:identifier>doi:10.1101/2021.11.08.467777</dc:identifier>
<dc:title><![CDATA[Prevalent mouse phenotypes in the unexplored druggable genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.08.467808v1?rss=1">
<title>
<![CDATA[
Microbial signals and lymphotoxin drive TNF-independent death of A20 and ABIN-1 deficient epithelium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.08.467808v1?rss=1</link>
<description><![CDATA[
Anti-TNF antibodies are effective for treating patients with inflammatory bowel disease (IBD), but many patients fail to respond to anti-TNF therapy, highlighting the importance of TNF-independent disease. We previously demonstrated that acute deletion of two IBD susceptibility genes, A20 (Tnfaip3) and Abin-1 (Tnip1), in intestinal epithelial cells (IECs) sensitizes mice to both TNF-dependent and TNF-independent death. Here we show that TNF-independent IEC death after A20 and Abin-1 deletion is rescued by germ-free derivation or deletion of MyD88, while deletion of Trif provides only partial protection. Combined deletion of Ripk3 and Casp8, which inhibits both apoptotic and necroptotic death, completely protects against death after acute deletion of A20 and Abin-1 in IECs. A20 and Abin-1-deficient IECs are sensitized to TNF-independent, TNFR-1-mediated death in response to lymphotoxin alpha (LT) homotrimers. Blockade of LT in vivo reduces weight loss and improves survival when combined with partial deletion of MyD88. These data show that microbial signals, MyD88, and LT all contribute to TNF-independent intestinal injury.

SUMMARYHere we show that germ-free derivation, MyD88 deletion, combined Ripk3 and Casp8 deletion, or anti-LT, all reduce TNF-independent intestinal injury after A20 and Abin-1 deletion.
]]></description>
<dc:creator>Rusu, I.</dc:creator>
<dc:creator>Mennillo, E.</dc:creator>
<dc:creator>Bain, J. L.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Ly, K.</dc:creator>
<dc:creator>Rosli, Y. Y.</dc:creator>
<dc:creator>Naser, M.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Advincula, R.</dc:creator>
<dc:creator>Achacoso, P.</dc:creator>
<dc:creator>Shao, L.</dc:creator>
<dc:creator>Razani, B.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Turnbaugh, J.</dc:creator>
<dc:creator>Turnbaugh, P.</dc:creator>
<dc:creator>Malynn, B. A.</dc:creator>
<dc:creator>Ma, A.</dc:creator>
<dc:creator>Kattah, M. G.</dc:creator>
<dc:date>2021-11-08</dc:date>
<dc:identifier>doi:10.1101/2021.11.08.467808</dc:identifier>
<dc:title><![CDATA[Microbial signals and lymphotoxin drive TNF-independent death of A20 and ABIN-1 deficient epithelium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.11.468235v1?rss=1">
<title>
<![CDATA[
Nucleolar-based Dux repression is essential for 2-cell stage exit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.11.468235v1?rss=1</link>
<description><![CDATA[
Upon fertilisation, the mammalian embryo must switch from dependence on maternal transcripts to transcribing its own genome, and in mice involves the transient upregulation of MERVL transposons and MERVL-driven genes at the 2-cell stage. The mechanisms and requirement for MERVL and 2-cell (2C) gene upregulation are poorly understood. Moreover, this MERVL-driven transcriptional program must be rapidly shut off to allow 2C exit and developmental progression. Here, we report that robust ribosomal RNA (rRNA) synthesis and nucleolar maturation are essential for exit from the 2C state. 2C-like cells and 2C embryos show similar immature nucleoli with altered structure and reduced rRNA output. We reveal that nucleolar disruption via blocking Pol I activity or preventing nucleolar phase separation enhances conversion to a 2C-like state in embryonic stem cells (ESCs) by detachment of the MERVL activator Dux from the nucleolar surface. In embryos, nucleolar disruption prevents proper Dux silencing and leads to 2-4 cell arrest. Our findings reveal an intriguing link between rRNA synthesis, nucleolar maturation and gene repression during early development.
]]></description>
<dc:creator>Xie, S. Q.</dc:creator>
<dc:creator>Leeke, B. J.</dc:creator>
<dc:creator>Whilding, C.</dc:creator>
<dc:creator>Wagner, R. T.</dc:creator>
<dc:creator>Garcia-Llagostera, F.</dc:creator>
<dc:creator>Chammas, P.</dc:creator>
<dc:creator>Cheung, N. T.-F.</dc:creator>
<dc:creator>Dormann, D.</dc:creator>
<dc:creator>McManus, M.</dc:creator>
<dc:creator>Percharde, M.</dc:creator>
<dc:date>2021-11-12</dc:date>
<dc:identifier>doi:10.1101/2021.11.11.468235</dc:identifier>
<dc:title><![CDATA[Nucleolar-based Dux repression is essential for 2-cell stage exit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.29.469773v1?rss=1">
<title>
<![CDATA[
The Astrin/SKAP Complex Reduces Friction at the Kinetochore-Microtubule Interface 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.29.469773v1?rss=1</link>
<description><![CDATA[
The kinetochore links chromosomes to spindle microtubules to drive chromosome segregation at cell division. While we know nearly all mammalian kinetochore proteins, how these give rise to the strong yet dynamic microtubule attachments required for function remains poorly understood. Here, we focus on the Astrin-SKAP complex, which localizes to bioriented kinetochores and is essential for chromosome segregation, but whose mechanical role is unclear. Live imaging reveals that SKAP depletion dampens movement and decreases coordination of metaphase sister kinetochores, and increases tension between them. Using laser ablation to isolate kinetochores bound to polymerizing vs depolymerizing microtubules, we show that without SKAP kinetochores move slower on both polymerizing and depolymerizing microtubules, and that more force is needed to rescue microtubules to polymerize. Thus, in contrast to previously described kinetochore proteins that increase grip on microtubules under force, Astrin-SKAP reduces grip, increasing attachment dynamics and force responsiveness and reducing friction. Together, our findings suggest a model where the Astrin-SKAP complex effectively "lubricates" correct, bioriented attachments to help preserve them.
]]></description>
<dc:creator>Rosas Salvans, M.</dc:creator>
<dc:creator>Sutanto, R.</dc:creator>
<dc:creator>Suresh, P.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2021-11-30</dc:date>
<dc:identifier>doi:10.1101/2021.11.29.469773</dc:identifier>
<dc:title><![CDATA[The Astrin/SKAP Complex Reduces Friction at the Kinetochore-Microtubule Interface]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.29.470434v1?rss=1">
<title>
<![CDATA[
Adolescent rats extend help to outgroup members, highlighting a neural network for group identity categorization. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.29.470434v1?rss=1</link>
<description><![CDATA[
Prosocial behavior, in particular helping others in need, occurs preferentially in response to the perceived distress of ones own group members, or ingroup. The development of neural mechanisms underlying social selectivity towards ingroup members are not well established. Here, we used a rat helping behavior test to explore the development and neural basis of ingroup bias for prosocial behavior in adolescent rats. We previously found that adult rats selectively help others from their own social group, and that this selectivity is associated with activation in reward and motivation circuits. Surprisingly, we found that adolescent rats helped both ingroup and outgroup members, evidence suggesting that ingroup bias emerges in adulthood. Analysis of brain-wide neural activity, indexed by expression of the early-immediate gene c-Fos, revealed increased activity for ingroup members across a broad set of regions, which was congruent for adults and adolescents. However, adolescents showed reduced hippocampal and insular activity, and increased orbitofrontal cortex activity compared to adults. Adolescent rats who did not help trapped others also demonstrated increased amygdala connectivity. Together, these findings demonstrate that biases for group-dependent prosocial behavior develop with age in rats and suggest that specific brain regions contribute to this prosocial selectivity, overall pointing to possible targets for the functional modulation of ingroup bias.

One Sentence SummaryProsocial selectivity increases with age in parallel with hippocampal and insular activation, providing insight into the neural classification of group membership.
]]></description>
<dc:creator>Breton, J.</dc:creator>
<dc:creator>Eisner, J. S.</dc:creator>
<dc:creator>Gandhi, V. S.</dc:creator>
<dc:creator>Musick, N.</dc:creator>
<dc:creator>Zhang, A.</dc:creator>
<dc:creator>Long, K. L. P.</dc:creator>
<dc:creator>Perloff, O. S.</dc:creator>
<dc:creator>Hu, K. Y.</dc:creator>
<dc:creator>Pham, C. M.</dc:creator>
<dc:creator>Lalchandani, P.</dc:creator>
<dc:creator>Barraza, M. K.</dc:creator>
<dc:creator>Kantor, B.</dc:creator>
<dc:creator>Kaufer, D.</dc:creator>
<dc:creator>Ben-Ami Bartal, I.</dc:creator>
<dc:date>2021-12-01</dc:date>
<dc:identifier>doi:10.1101/2021.11.29.470434</dc:identifier>
<dc:title><![CDATA[Adolescent rats extend help to outgroup members, highlighting a neural network for group identity categorization.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.30.470624v1?rss=1">
<title>
<![CDATA[
15N metabolic labeling quantification workflow in Arabidopsis using Protein Prospector 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.30.470624v1?rss=1</link>
<description><![CDATA[
Metabolic labeling using stable isotopes is widely used for the relative quantification of proteins in proteomic studies. In plants, metabolic labeling using 15N has great potential, but the associated complexity of data analysis has limited its usage. Here, we present the 15N stable-isotope labeled protein quantification workflow utilizing open-access web-based software Protein Prospector. Further, we discuss several important features of 15N labeling required to make reliable and precise protein quantification. These features include ratio adjustment based on labeling efficiency, median and interquartile range for protein ratios, isotope cluster pattern matching to flag incorrect monoisotopic peak assignment, and caching of quantification results for fast retrieval.
]]></description>
<dc:creator>Shrestha, R.</dc:creator>
<dc:creator>Reyes, A. V.</dc:creator>
<dc:creator>Baker, P. R.</dc:creator>
<dc:creator>Wang, Z.-Y.</dc:creator>
<dc:creator>Chalkley, R. J.</dc:creator>
<dc:creator>Xu, S.-L.</dc:creator>
<dc:date>2021-12-01</dc:date>
<dc:identifier>doi:10.1101/2021.11.30.470624</dc:identifier>
<dc:title><![CDATA[15N metabolic labeling quantification workflow in Arabidopsis using Protein Prospector]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.01.470575v1?rss=1">
<title>
<![CDATA[
Input-specific control of interneuron numbers in nascent striatal networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.01.470575v1?rss=1</link>
<description><![CDATA[
The assembly of functional neuronal circuits requires appropriate numbers of distinct classes of neurons, but the mechanisms through which their relative proportions are established remain poorly defined. Investigating the mouse striatum, here we found that the two most prominent subtypes of striatal interneurons, parvalbumin-expressing (PV+) GABAergic and cholinergic (ChAT+) interneurons, undergo extensive programmed cell death between the first and second postnatal weeks. Remarkably, the survival of PV+ and ChAT+ interneurons is regulated by distinct mechanisms mediated by their specific afferent connectivity. While long-range cortical inputs control PV+ interneuron survival, ChAT+ interneuron survival is regulated by local input from the medium spiny neurons. Our results identify input-specific circuit mechanisms that operate during the period of programmed cell death to establish the final number of interneurons in nascent striatal networks.
]]></description>
<dc:creator>Sreenivasan, V.</dc:creator>
<dc:creator>Serafeimidou-Pouliou, E.</dc:creator>
<dc:creator>Exposito-Alonso, D.</dc:creator>
<dc:creator>Bercsenyi, K.</dc:creator>
<dc:creator>Bernard, C.</dc:creator>
<dc:creator>Bae, S.-E.</dc:creator>
<dc:creator>Oozeer, F.</dc:creator>
<dc:creator>Hanusz-Godoy, A.</dc:creator>
<dc:creator>Edwards, R.</dc:creator>
<dc:creator>Marin, O.</dc:creator>
<dc:date>2021-12-02</dc:date>
<dc:identifier>doi:10.1101/2021.12.01.470575</dc:identifier>
<dc:title><![CDATA[Input-specific control of interneuron numbers in nascent striatal networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.01.470846v1?rss=1">
<title>
<![CDATA[
Application of Parallel Reaction Monitoring in 15N labeled Samples for Quantification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.01.470846v1?rss=1</link>
<description><![CDATA[
Accurate relative quantification is critical in proteomic studies. The incorporation of stable isotope 15N to plant-expressed proteins in vivo is a powerful tool for accurate quantification with a major advantage of reducing preparative and analytical variabilities. However, 15N labeling quantification has several challenges. Less identifications are often observed in the heavy labeled samples because of incomplete labeling, resulting in missing values in reciprocal labeling experiments. Inaccurate quantification can happen when there is contamination from co-eluting peptides or chemical noise in the MS1 survey scan. These drawbacks in quantification can be more pronounced in less abundant but biologically interesting proteins, which often have very few identified peptides. Here we demonstrate the application of parallel reaction monitoring (PRM) to 15N labeled samples on a high resolution, high mass accuracy Orbitrap mass spectrometer to achieve reliable quantification even of low abundance proteins in samples.
]]></description>
<dc:creator>Reyes, A. V.</dc:creator>
<dc:creator>Shrestha, R.</dc:creator>
<dc:creator>Baker, P. R.</dc:creator>
<dc:creator>Chalkley, R. J.</dc:creator>
<dc:creator>Xu, S.-L.</dc:creator>
<dc:date>2021-12-03</dc:date>
<dc:identifier>doi:10.1101/2021.12.01.470846</dc:identifier>
<dc:title><![CDATA[Application of Parallel Reaction Monitoring in 15N labeled Samples for Quantification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.05.471221v1?rss=1">
<title>
<![CDATA[
Control of nuclear size by osmotic forces in Schizosaccharomyces pombe 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.05.471221v1?rss=1</link>
<description><![CDATA[
The size of the nucleus scales robustly with cell size so that the nuclear-to-cell volume ratio (N/C ratio) is maintained during cell growth in many cell types. The mechanism responsible for this scaling remains mysterious. Previous studies have established that the N/C ratio is not determined by DNA amount but is instead influenced by factors such as nuclear envelope mechanics and nuclear transport. Here, we developed a quantitative model for nuclear size control based upon colloid osmotic pressure and tested key predictions in the fission yeast Schizosaccharomyces pombe. This model posits that the N/C ratio is determined by the numbers of macromolecules in the nucleoplasm and cytoplasm. Osmotic shift experiments showed that the fission yeast nucleus behaves as an ideal osmometer whose volume is primarily dictated by osmotic forces. Inhibition of nuclear export caused accumulation of macromolecules and an increase in crowding in the nucleoplasm, leading to nuclear swelling. We further demonstrated that the N/C ratio is maintained by a homeostasis mechanism based upon synthesis of macromolecules during growth. These studies demonstrate the functions of colloid osmotic pressure in intracellular organization and size control.
]]></description>
<dc:creator>Lemiere, J.</dc:creator>
<dc:creator>Real-Calderon, P.</dc:creator>
<dc:creator>Holt, L.</dc:creator>
<dc:creator>Fai, T.</dc:creator>
<dc:creator>Chang, F.</dc:creator>
<dc:date>2021-12-07</dc:date>
<dc:identifier>doi:10.1101/2021.12.05.471221</dc:identifier>
<dc:title><![CDATA[Control of nuclear size by osmotic forces in Schizosaccharomyces pombe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.10.472156v1?rss=1">
<title>
<![CDATA[
Single-fiber nucleosome density shapes the regulatory output of a mammalian chromatin remodeling enzyme 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.10.472156v1?rss=1</link>
<description><![CDATA[
ATP-dependent chromatin remodelers regulate the DNA accessibility required of virtually all nuclear processes. Biochemical studies have provided insight into remodeler action at the nucleosome level, but how these findings translate to activity on chromatin fibers in vitro and in vivo remains poorly understood. Here, we present a massively multiplex single-molecule platform allowing high-resolution mapping of nucleosomes on fibers assembled on mammalian genomic sequences. We apply this method to distinguish between competing models for chromatin remodeling by the essential ISWI ATPase SNF2h: linker-length-dependent dynamic positioning versus fixed-linker-length static clamping. Our single-fiber data demonstrate that SNF2h operates as a density-dependent, length-sensing chromatin remodeler whose ability to decrease or increase DNA accessibility depends on single-fiber nucleosome density. In vivo, this activity manifests as different regulatory modes across epigenomic domains: at canonically-defined heterochromatin, SNF2h generates evenly-spaced nucleosome arrays of multiple nucleosome repeat lengths; at SNF2h-dependent accessible sites, SNF2h slides nucleosomes to increase accessibility of motifs for the essential transcription factor CTCF. Overall, our generalizable approach provides molecularly-precise views of the processes that shape nuclear physiology. Concurrently, our data illustrate how a mammalian chromatin remodeling enzyme can effectively sense nucleosome density to induce diametrically-opposed regulatory effects within the nucleus.
]]></description>
<dc:creator>Abdulhay, N. J.</dc:creator>
<dc:creator>Hsieh, L. J.</dc:creator>
<dc:creator>McNally, C. P.</dc:creator>
<dc:creator>Ketavarapu, M.</dc:creator>
<dc:creator>Kasinathan, S.</dc:creator>
<dc:creator>Nanda, A. S.</dc:creator>
<dc:creator>Ostrowski, M. S.</dc:creator>
<dc:creator>Wu, K.</dc:creator>
<dc:creator>Moore, C. M.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:date>2021-12-11</dc:date>
<dc:identifier>doi:10.1101/2021.12.10.472156</dc:identifier>
<dc:title><![CDATA[Single-fiber nucleosome density shapes the regulatory output of a mammalian chromatin remodeling enzyme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.16.473013v1?rss=1">
<title>
<![CDATA[
Mapping information-rich genotype-phenotype landscapes with genome-scale Perturb-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.16.473013v1?rss=1</link>
<description><![CDATA[
A central goal of genetics is to define the relationships between genotypes and phenotypes. High-content phenotypic screens such as Perturb-seq (pooled CRISPR-based screens with single-cell RNA-sequencing readouts) enable massively parallel functional genomic mapping but, to date, have been used at limited scales. Here, we perform genome-scale Perturb-seq targeting all expressed genes with CRISPR interference (CRISPRi) across >2.5 million human cells and present a framework to power biological discovery with the resulting genotype-phenotype map. We use transcriptional phenotypes to predict the function of poorly-characterized genes, uncovering new regulators of ribosome biogenesis (including CCDC86, ZNF236, and SPATA5L1), transcription (C7orf26), and mitochondrial respiration (TMEM242). In addition to assigning gene function, single-cell transcriptional phenotypes allow for in-depth dissection of complex cellular phenomena - from RNA processing to differentiation. We leverage this ability to systematically identify the genetic drivers and consequences of aneuploidy and to discover an unanticipated layer of stress-specific regulation of the mitochondrial genome. Our information-rich genotype-phenotype map reveals a multidimensional portrait of gene function and cellular behavior.
]]></description>
<dc:creator>Replogle, J. M.</dc:creator>
<dc:creator>Saunders, R. A.</dc:creator>
<dc:creator>Pogson, A. N.</dc:creator>
<dc:creator>Hussmann, J. A.</dc:creator>
<dc:creator>Lenail, A.</dc:creator>
<dc:creator>Guna, A.</dc:creator>
<dc:creator>Mascibroda, L.</dc:creator>
<dc:creator>Wagner, E. J.</dc:creator>
<dc:creator>Adelman, K.</dc:creator>
<dc:creator>Bonnar, J. L.</dc:creator>
<dc:creator>Jost, M.</dc:creator>
<dc:creator>Norman, T. M.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:date>2021-12-17</dc:date>
<dc:identifier>doi:10.1101/2021.12.16.473013</dc:identifier>
<dc:title><![CDATA[Mapping information-rich genotype-phenotype landscapes with genome-scale Perturb-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.16.473030v1?rss=1">
<title>
<![CDATA[
Modeling the dynamics of within-host viral infection and evolution predicts quasispecies distributions and phase boundaries separating distinct classes of infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.16.473030v1?rss=1</link>
<description><![CDATA[
We use computational modeling to study within-host viral infection and evolution. In our model, viruses exhibit variable binding to cells, with better infection and replication countered by a stronger immune response and a high rate of mutation. By varying host conditions (permissivity to viral entry T and immune clearance intensity A) for large numbers of cells and viruses, we study the dynamics of how viral populations evolve from initial infection to steady state and obtain a phase diagram of the range of cell and viral responses. We find three distinct replicative strategies corresponding to three physiological classes of viral infections: acute, chronic, and opportunistic. We show similarities between our findings and the behavior of real viral infections such as common flu, hepatitis, and SARS-CoV-2019. The phases associated with the three strategies are separated by a phase transition of primarily first order, in addition to a crossover region. Our simulations also reveal a wide range of physical phenomena, including metastable states, periodicity, and glassy dynamics. Lastly, our results suggest that the resolution of acute viral disease in patients whose immunity cannot be boosted can only be achieved by significant inhibition of viral infection and replication.

Author summaryVirus, in particular RNA viruses, often produce offspring with slightly altered genetic composition. This process occurs both across host populations and within a single host over time. Here, we study the interactions of viruses with cells inside a host over time. In our model, the viruses encounter host cell defenses characterized by two parameters: permissivity to viral entry T and immune response A). The viruses then mutate upon reproduction, eventually resulting in a distribution of related viral types termed a quasi-species distribution. Across varying host conditions (T, A), three distinct viral quasi-species types emerge over time, corresponding to three classes of viral infections: acute, chronic and opportunistic. We interpret these results in terms of real viral types such as common flu, hepatitis, and also SARS-CoV-2019. Analysis of viral of viral mutant populations over a wide range of permissivity and immunity, for large numbers of cells and viruses, reveals phase transitions that separate the three classes of viruses, both in the infection-cycle dynamics and at steady state. We believe that such a multiscale approach for the study of within-host viral infections, spanning individual proteins to collections of cells, can provide insight into developing more effective therapies for viral disease.
]]></description>
<dc:creator>Lewis, G. R.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:creator>Jones, B. A.</dc:creator>
<dc:date>2021-12-20</dc:date>
<dc:identifier>doi:10.1101/2021.12.16.473030</dc:identifier>
<dc:title><![CDATA[Modeling the dynamics of within-host viral infection and evolution predicts quasispecies distributions and phase boundaries separating distinct classes of infections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.16.473055v1?rss=1">
<title>
<![CDATA[
ICoRD: Iterative Correlation-Based ROI Detection Method for the Extraction of Neural Signals in Calcium Imaging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.16.473055v1?rss=1</link>
<description><![CDATA[
In vivo calcium imaging is a standard neuroimaging technique that allows selective observation of target neuronal activities. In calcium imaging, neuron activation signals provide key information for the investigation of neural circuits. For efficient extraction of the calcium signals of neurons, selective detection of the region of interest (ROI) pixels corresponding to the active subcellular region of the target neuron is essential. However, current ROI detection methods for calcium imaging data exhibit a relatively low signal extraction performance from neurons with a low signal-to-noise power ratio (SNR). This is problematic because a low SNR is unavoidable in many biological experiments. Therefore, we propose an iterative correlation-based ROI detection (ICoRD) method that robustly extracts the calcium signal of the target neuron from a calcium imaging series with severe noise. ICoRD extracts calcium signals closer to the ground-truth calcium signal than the conventional method from simulated calcium imaging data in all low SNR ranges. Additionally, this study confirmed that ICoRD robustly extracts activation signals against noise, even within in vivo environments. ICoRD showed reliable detection from neurons with a low SNR and sparse activation, which were not detected by conventional methods. ICoRD will facilitate our understanding of neural circuit activity by providing significantly improved ROI detection in noisy images.
]]></description>
<dc:creator>Kang, S.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Lim, S.-H.</dc:creator>
<dc:creator>Choi, J. H.</dc:creator>
<dc:creator>Rah, J.-C.</dc:creator>
<dc:creator>Choi, J.-W.</dc:creator>
<dc:date>2021-12-17</dc:date>
<dc:identifier>doi:10.1101/2021.12.16.473055</dc:identifier>
<dc:title><![CDATA[ICoRD: Iterative Correlation-Based ROI Detection Method for the Extraction of Neural Signals in Calcium Imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.21.473768v1?rss=1">
<title>
<![CDATA[
Combination of Mycobacterium tuberculosis RS ratio and CFU improves the ability of murine efficacy experiments to distinguish between drug treatments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.21.473768v1?rss=1</link>
<description><![CDATA[
Murine tuberculosis drug efficacy studies have historically monitored bacterial burden based on colony forming units of M. tuberculosis in lung homogenate. In an alternative approach, a recently described molecular pharmacodynamic marker called the RS ratio quantifies drug effect on a fundamental cellular process: ongoing ribosomal RNA synthesis. Here we evaluated the ability of different pharmacodynamic markers to distinguish between treatments in three BALB/c mouse experiments at two institutions. We confirmed that different pharmacodynamic markers measure distinct biological responses. We found that a combination of pharmacodynamic markers distinguishes between treatments better than any single marker. The combination of the RS ratio with colony forming units showed the greatest ability to recapitulate the rank order of regimen treatment-shortening activity, providing proof of concept that simultaneous assessment of pharmacodynamic markers measuring different properties will enhance insight gained from animal models and accelerate development of new combination regimens. These results suggest potential for a new era in which antimicrobial therapies are evaluated not only on culture-based measures of bacterial burden but also on molecular assays that indicate how drugs impact the physiological state of the pathogen.
]]></description>
<dc:creator>Dide-Agossou, C.</dc:creator>
<dc:creator>Bauman, A. A.</dc:creator>
<dc:creator>Ramey, M. E.</dc:creator>
<dc:creator>Rossmassler, K.</dc:creator>
<dc:creator>Al Mubarak, R.</dc:creator>
<dc:creator>Pauly, S.</dc:creator>
<dc:creator>Voskuil, M. I.</dc:creator>
<dc:creator>Garcia?Cremades, M.</dc:creator>
<dc:creator>Savic, R. M.</dc:creator>
<dc:creator>Nahid, P.</dc:creator>
<dc:creator>Moore, C. M.</dc:creator>
<dc:creator>Tasneen, R.</dc:creator>
<dc:creator>Nuermburger, E.</dc:creator>
<dc:creator>Robertson, G. T.</dc:creator>
<dc:creator>Walter, N. D.</dc:creator>
<dc:date>2021-12-24</dc:date>
<dc:identifier>doi:10.1101/2021.12.21.473768</dc:identifier>
<dc:title><![CDATA[Combination of Mycobacterium tuberculosis RS ratio and CFU improves the ability of murine efficacy experiments to distinguish between drug treatments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.30.474394v1?rss=1">
<title>
<![CDATA[
P-TEFb is degraded by Siah1/2 in quiescent cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.30.474394v1?rss=1</link>
<description><![CDATA[
P-TEFb, composed of CycT1 and CDK9, regulates the elongation of transcription by RNA polymerase II. In proliferating cells, it is regulated by 7SK snRNA in the 7SK snRNP complex. In resting cells, P-TEFb is absent, because CycT1 is dephosphorylated, released from CDK9 and rapidly degraded. In this study, we identified the mechanism of this degradation. We mapped the ubiquitination and degradation of free CycT1 to its N-terminal region from positions 1 to 280. This region is ubiquitinated at six lysines, where E3 ligases Siah1 and Siah2 bind and degrade these sequences. Importantly, the inhibition of Siah1/2 rescued the expression of free CycT1 in proliferating as well as resting primary cells. We conclude that Siah1/2 are the E3 ligases that bind and degrade the dissociated CycT1 in resting, terminally differentiated, anergic and/or exhausted cells.
]]></description>
<dc:creator>Huang, F.</dc:creator>
<dc:creator>Feng, Y.</dc:creator>
<dc:creator>Peterlin, B. M.</dc:creator>
<dc:creator>Fujinaga, K.</dc:creator>
<dc:date>2022-01-01</dc:date>
<dc:identifier>doi:10.1101/2021.12.30.474394</dc:identifier>
<dc:title><![CDATA[P-TEFb is degraded by Siah1/2 in quiescent cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.04.474746v1?rss=1">
<title>
<![CDATA[
hPSC-Derived Enteric Ganglioids Model Human ENS Development and Function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.04.474746v1?rss=1</link>
<description><![CDATA[
The enteric nervous system (ENS) plays a central role in gut physiology and mediating the crosstalk between the gastrointestinal (GI) tract and other organs. The human ENS has remained elusive, highlighting the need for an in vitro modeling and mapping blueprint. Here we map out the developmental and functional features of the human ENS, by establishing robust and scalable 2D ENS cultures and 3D enteric ganglioids from human pluripotent stem cells (hPSCs). These models recapitulate the remarkable neuronal and glial diversity found in primary tissue and enable comprehensive molecular analyses that uncover functional and developmental relationships within these lineages. As a salient example of the power of this system, we performed in-depth characterization of enteric nitrergic neurons (NO neurons) which are implicated in a wide range of GI motility disorders. We conducted an unbiased screen and identified drug candidates that modulate the activity of NO neurons and demonstrated their potential in promoting motility in mouse colonic tissue ex vivo. We established a high-throughput strategy to define the developmental programs involved in NO neuron specification and discovered that PDGFR inhibition boosts the induction of NO neurons in enteric ganglioids. Transplantation of these ganglioids in the colon of NO neuron-deficient mice results in extensive tissue engraftment, providing a xenograft model for the study of human ENS in vivo and the development of cell-based therapies for neurodegenerative GI disorders. These studies provide a framework for deciphering fundamental features of the human ENS and designing effective strategies to treat enteric neuropathies.
]]></description>
<dc:creator>Majd, H.</dc:creator>
<dc:creator>Samuel, R. M.</dc:creator>
<dc:creator>Ramirez, J. T.</dc:creator>
<dc:creator>Kalantari, A.</dc:creator>
<dc:creator>Barber, K.</dc:creator>
<dc:creator>Ghazizadeh, Z.</dc:creator>
<dc:creator>Chemel, A. K.</dc:creator>
<dc:creator>Cesiulis, A.</dc:creator>
<dc:creator>Richter, M. N.</dc:creator>
<dc:creator>Das, S.</dc:creator>
<dc:creator>Keefe, M. G.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Shiv, R.</dc:creator>
<dc:creator>McCann, C. J.</dc:creator>
<dc:creator>Bhat, S.</dc:creator>
<dc:creator>Khoroshkin, M.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Thapar, N.</dc:creator>
<dc:creator>Kaltschmidt, J. A.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:date>2022-01-04</dc:date>
<dc:identifier>doi:10.1101/2022.01.04.474746</dc:identifier>
<dc:title><![CDATA[hPSC-Derived Enteric Ganglioids Model Human ENS Development and Function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.04.474985v1?rss=1">
<title>
<![CDATA[
Exploring the rules of chimeric antigen receptor phenotypic output using combinatorial signaling motif libraries and machine learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.04.474985v1?rss=1</link>
<description><![CDATA[
Chimeric antigen receptor (CAR) costimulatory domains steer the phenotypic output of therapeutic T cells. In most cases these domains are derived from native immune receptors, composed of signaling motif combinations selected by evolution. To explore if non-natural combinations of signaling motifs could drive novel cell fates of interest, we constructed a library of CARs containing [~]2,300 synthetic costimulatory domains, built from combinations of 13 peptide signaling motifs. The library produced CARs driving diverse fate outputs, which were sensitive to motif combinations and configurations. Neural networks trained to decode the combinatorial grammar of CAR signaling motifs allowed extraction of key design rules. For example, the non-native combination of TRAF- and PLC{gamma}1-binding motifs was found to simultaneously enhance cytotoxicity and stemness, a clinically desirable phenotype associated with effective and durable tumor killing. The neural network accurately predicts that addition of PLC{gamma}1-binding motifs improves this phenotype when combined with TRAF-binding motifs, but not when combined with other immune signaling motifs (e.g. PI3K-or Grb2-binding motifs). This work shows how libraries built from the minimal building blocks of signaling, combined with machine learning, can efficiently guide engineering of receptors with desired phenotypes.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=60 SRC="FIGDIR/small/474985v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Daniels, K. G.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Simic, M. S.</dc:creator>
<dc:creator>Bhargava, H. K.</dc:creator>
<dc:creator>Capponi, S.</dc:creator>
<dc:creator>Tonai, Y.</dc:creator>
<dc:creator>Yu, W.</dc:creator>
<dc:creator>Bianco, S.</dc:creator>
<dc:creator>Lim, W. A.</dc:creator>
<dc:date>2022-01-04</dc:date>
<dc:identifier>doi:10.1101/2022.01.04.474985</dc:identifier>
<dc:title><![CDATA[Exploring the rules of chimeric antigen receptor phenotypic output using combinatorial signaling motif libraries and machine learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.06.475287v1?rss=1">
<title>
<![CDATA[
Proteomic Dissection of a Giant Cell 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.06.475287v1?rss=1</link>
<description><![CDATA[
Many individual proteins have been identified as having defined positions relative to cell polarity axes, raising the question of what fraction of all proteins may have polarized localizations. We took advantage of the giant ciliate Stentor coeruleus to quantify the extent of polarized localization proteome-wide. This trumpet-shaped unicellular organism shows a clear morphological anterior-posterior axis defined by a circular array of cilia known as a membranellar band at one end, and a holdfast at the other end. Because individual Stentor cells are over a millimeter in length, we were able to cut the cells into three pieces along the anterior-posterior axis, followed by proteomic analysis of proteins enriched in each piece. We find that approximately 30% of all detected proteins show a polarized location relative to the anterior-posterior cell axis. Proteins with polarized enrichment include centrin-like proteins, calcium-regulated kinases, orthologs of SFI1 and GAS2, and proteases. At the organelle level, nuclear and mitochondrial proteins are enriched in the anterior half of the cell body, but not in the membranellar band itself, while ribosome related proteins are apparently uniformly distributed. RNAi of signaling proteins enriched in the membranellar band, which is the anterior-most structure in the cell, revealed a protein phosphatase 2 subunit b ortholog required for closure of the membranellar band into the ring shape characteristic of Stentor. These results suggest that a large fraction of the Stentor proteome has a polarized localization, and provide a protein-level framework for future analysis of pattern formation and regeneration in Stentor as well as defining a general strategy for subcellular spatial proteomics based on physical dissection of cells.
]]></description>
<dc:creator>Lin, A.</dc:creator>
<dc:creator>Piehowski, P.</dc:creator>
<dc:creator>Tsai, C.-F.</dc:creator>
<dc:creator>Makushok, T.</dc:creator>
<dc:creator>Yi, L.</dc:creator>
<dc:creator>Diaz, U.</dc:creator>
<dc:creator>Yan, C.</dc:creator>
<dc:creator>Summers, D.</dc:creator>
<dc:creator>Sood, P.</dc:creator>
<dc:creator>Smith, R.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2022-01-06</dc:date>
<dc:identifier>doi:10.1101/2022.01.06.475287</dc:identifier>
<dc:title><![CDATA[Proteomic Dissection of a Giant Cell]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.13.476250v1?rss=1">
<title>
<![CDATA[
Naive arthritogenic SKG T cells have a defect in anergy and a repertoire pruned by superantigen 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.13.476250v1?rss=1</link>
<description><![CDATA[
How autoreactive CD4 T cells develop to cause rheumatoid arthritis remains unknown. We used a reporter for antigen-receptor signaling in the SKG autoimmune arthritis model to profile a T cell subpopulation enriched for arthritogenic naive CD4 T cells before arthritis onset by bulk and single cell RNA and T cell antigen-receptor (TCR) sequencing. Our analyses reveal that despite their impaired proximal TCR signaling, a subset of SKG naive CD4 T cells that have recently encountered endogenous antigen upregulate gene programs associated with positive regulation of T cell activation and cytokine signaling at higher levels than wild type cells in the pre-disease state. These arthritogenic cells also induce genes associated with negative regulation of T cell activation but do so less efficiently than wild type cells. Furthermore, their TCR sequences exhibit a previously unrecognized biased peripheral TCR V{beta} repertoire likely driven by endogenous viral superantigens. These particular V{beta}s, known to recognize endogenous mouse mammary tumor virus (MMTV) superantigen, are further expanded in arthritic joints. Our results demonstrate that autoreactive naive CD4 T cells which recognize endogenous viral superantigens are poised to cause disease by their altered transcriptome.

Summary blurbSelf-reactive SKG T cells that escaped negative selection harbor an independent defect in anergy that, together with chronic antigen stimulation, sets the stage for disease. Moreover, we propose a novel role for endogenous mouse mammary tumor virus (MMTV) superantigen in promoting arthritogenic T cell responses.
]]></description>
<dc:creator>Ashouri, J. F.</dc:creator>
<dc:creator>McCarthy, E.</dc:creator>
<dc:creator>Yu, S.</dc:creator>
<dc:creator>Perlmutter, N.</dc:creator>
<dc:creator>Lin, C.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Weiss, A.</dc:creator>
<dc:date>2022-01-16</dc:date>
<dc:identifier>doi:10.1101/2022.01.13.476250</dc:identifier>
<dc:title><![CDATA[Naive arthritogenic SKG T cells have a defect in anergy and a repertoire pruned by superantigen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.19.476699v1?rss=1">
<title>
<![CDATA[
STAT3 regulates cytokine production downstream of TNFR1 in part by inducing expression of TNFAIP3/A20 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.19.476699v1?rss=1</link>
<description><![CDATA[
BackgroundPrevious work demonstrated that the Signal Transducer and Activator of Transcription 3 (STAT3) is activated downstream of the Type 1 TNF Receptor. However, whether and how STAT3 regulates gene expression downstream of TNFR1 has not been elucidated.

MethodsGlobal transcriptome analysis by RNA sequencing was performed in wild type and STAT3 knockout mouse embryonic fibroblasts (MEFs) stimulated with TNF. The fold changes in gene expression were assessed bioinformatically. Results of the RNA sequencing were validated at the protein level by using multiplex cytokine assays and immunoblotting.

ResultsStimulation of MEFs with TNF or an agonist antibody to TNFR1 activated STAT3, and this was inhibited by pharmacological inhibition of Jak2 and cSrc. At 4 hours after TNF stimulation, STAT3 knockout MEFs had a greater level than WT MEFs of induction of the chemokines Ccl2, Cxcl1 and Cxcl10 at the RNA and protein levels. Mechanistically, this was due STAT3 promoting the expression of Tnfaip3/A20, a ubiquitin modifying enzyme that inhibits inflammation, in wild-type MEFs at early timepoints after TNFR1 stimulation. In STAT3 knockout MEFs TNF failed to induce the expression of Tnfaip3/A20 or GM-CSF when acting through TNFR1. Expression of A20 into STAT3 knockout MEFs suppressed cytokine expression.

ConclusionSTAT3 limits the induction of Ccl2, Cxcl1 and Cxcl10 in response to TNFR1 activation by promoting the expression of Tnfaip3/A20. On the other hand, STAT3 promotes the expression of GM-CSF in response to TNFR1 stimulation. These results show that STAT3 modulates inflammatory signaling by TNF in normal cells.
]]></description>
<dc:creator>Antonia, R. J.</dc:creator>
<dc:creator>Karelehto, E.</dc:creator>
<dc:creator>Toriguchi, K.</dc:creator>
<dc:creator>Matli, M.</dc:creator>
<dc:creator>Warren, R. S.</dc:creator>
<dc:creator>Pfeffer, L. M.</dc:creator>
<dc:creator>Donner, D. B.</dc:creator>
<dc:date>2022-01-20</dc:date>
<dc:identifier>doi:10.1101/2022.01.19.476699</dc:identifier>
<dc:title><![CDATA[STAT3 regulates cytokine production downstream of TNFR1 in part by inducing expression of TNFAIP3/A20]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.19.476935v1?rss=1">
<title>
<![CDATA[
Therapeutic Interruption of T Cell Development Generates High-Affinity T Cells That Escape Exhaustion and Improve Cancer Immunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.19.476935v1?rss=1</link>
<description><![CDATA[
Availability of effective anti-tumor T cells is limited by cancer immunoediting, which depletes neoantigens, and central tolerance, which eliminates developing T cells with high-affinity T cell receptors (TCRs) against tumor self-antigens. Remaining tumor-reactive T cells are often exhausted after immune checkpoint blockade (ICB). Whether endogenous T cells with high- affinity TCRs against tumor self-antigens can be generated to circumvent exhaustion and reject neoantigen-poor tumors is unclear. We show that transiently interrupting central tolerance through RANKL blockade unleashes T cells possessing TCRs with self-reactive features that enable ICB to reject poorly immunogenic tumors. Upon recognition of tumor self-antigens, these T cells exhibit enhanced TCR signaling, enrichment in NFAT/AP-1 genes, and lymph node priming. Consequently, memory-precursor T cells against tumor self-antigens are generated, avoid exhaustion, and become effector-memory cells with transcriptional features associated with clinical ICB response. Thus, interrupting central tolerance provides T cells with tumor- directed autoreactivity that avoid exhaustion and improve immunotherapy.
]]></description>
<dc:creator>Dhuey, E.</dc:creator>
<dc:creator>Oldridge, O.</dc:creator>
<dc:creator>Ravishankar, R.</dc:creator>
<dc:creator>Dada, H.</dc:creator>
<dc:creator>Yu, Y.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>Minn, A. J.</dc:creator>
<dc:date>2022-01-21</dc:date>
<dc:identifier>doi:10.1101/2022.01.19.476935</dc:identifier>
<dc:title><![CDATA[Therapeutic Interruption of T Cell Development Generates High-Affinity T Cells That Escape Exhaustion and Improve Cancer Immunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.28.478269v1?rss=1">
<title>
<![CDATA[
Synthetic glycans that control gut microbiome structure mitigate colitis in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.28.478269v1?rss=1</link>
<description><![CDATA[
Relative abundances of bacterial species in the gut microbiome have been linked to many diseases. Species of gut bacteria are ecologically differentiated by their abilities to metabolize different glycans, making glycan delivery a powerful way to alter the microbiome to promote health. We describe the properties and therapeutic potential of chemically diverse synthetic glycans (SGs). Fermentation of SGs by gut microbiome cultures resulted in compound-specific shifts in taxonomic and metabolite profiles not observed with reference glycans, including prebiotics. Model enteric pathogens grow poorly on most SGs, potentially increasing their safety for at-risk populations. SGs increased survival, reduced weight loss, and improved clinical scores in mouse models of colitis. Synthetic glycans are thus a promising modality to improve health through selective changes to the gut microbiome.
]]></description>
<dc:creator>Tolonen, A. C.</dc:creator>
<dc:creator>Beauchemin, N.</dc:creator>
<dc:creator>Bayne, C.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Tan, J.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Meehan, B.</dc:creator>
<dc:creator>Meisner, J.</dc:creator>
<dc:creator>Millet, Y.</dc:creator>
<dc:creator>Rapp, E.</dc:creator>
<dc:creator>Kottler, R.</dc:creator>
<dc:creator>Murphy, C.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:creator>von Maltzahn, G.</dc:creator>
<dc:creator>Liu, C. M.</dc:creator>
<dc:creator>van Hylckama Vlieg, J.</dc:creator>
<dc:date>2022-01-29</dc:date>
<dc:identifier>doi:10.1101/2022.01.28.478269</dc:identifier>
<dc:title><![CDATA[Synthetic glycans that control gut microbiome structure mitigate colitis in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.30.478390v1?rss=1">
<title>
<![CDATA[
Clemizole and Trazodone are Effective Antiseizure Treatments in a Zebrafish Model of STXBP1 Disorder 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.30.478390v1?rss=1</link>
<description><![CDATA[
ObjectiveCRISPR-Cas9-generated zebrafish carrying a 12 base-pair deletion in stxbpb1b, a paralog sharing 79% amino acid sequence identity with human, exhibit spontaneous electrographic seizures during larval stages of development. Zebrafish stxbp1b mutants provide an efficient preclinical platform to test antiseizure therapeutics. The present study was designed to test prototype antiepileptic drugs approved for clinical use and two recently identified repurposed drugs with antiseizure activity.

MethodsLarval homozygous stxbp1b zebrafish (4 days post-fertilization) were agarose-embedded and monitored for electrographic seizure activity using a local field recording electrode placed in midbrain. Frequency of ictal-like events was evaluated at baseline and following 45 min of continuous drug exposure (1 mM, bath application). Analysis was performed on coded files by an experimenter blinded to drug treatment and genotype.

ResultsPhenytoin, valproate, ethosuximide, levetiracetam, and diazepam had no effect on ictal-like event frequency in stxbp1b mutant zebrafish. Clemizole and trazodone decreased ictal-like event frequency in stxbp1b mutant zebrafish by 80% and 83%, respectively. These results suggest that repurposed drugs with serotonin receptor binding affinities could be effective antiseizure treatments.

SignificanceClemizole and trazodone were identified in a larval zebrafish model for Dravet syndrome. Based primarily on these preclinical zebrafish studies, compassionate-use and double-blind clinical trials with both drugs have progressed. The present study extends this approach to a preclinical zebrafish model representing STXBP1-related disorders, and suggests that future clinical studies may be warranted.
]]></description>
<dc:creator>Moog, M.</dc:creator>
<dc:creator>Baraban, S. C.</dc:creator>
<dc:date>2022-01-30</dc:date>
<dc:identifier>doi:10.1101/2022.01.30.478390</dc:identifier>
<dc:title><![CDATA[Clemizole and Trazodone are Effective Antiseizure Treatments in a Zebrafish Model of STXBP1 Disorder]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.07.479384v1?rss=1">
<title>
<![CDATA[
Influence of earlier human activity on gut microbiota and immune responses of Darwin's finches in the Galapagos Islands 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.07.479384v1?rss=1</link>
<description><![CDATA[
Urbanization can influence animal traits, including immunity and gut microbiota. Over the past several decades, the Galapagos Islands have seen rapid resident human population growth and tourist activity, leading to varying levels of human activity across Islands. Consequently, diet, gut microbiota, and immunity of endemic animals, such as Darwins finches, may have changed. The goal of this study was to determine the effect of land use on the immune response, gut microbiota, and body measurements of Darwins finches in 2008, at a time of rapidly increasing human activity in the Islands. Specifically, we compared proxies of immunity (lysozyme activity, and haptoglobin, complement antibody, and natural antibody levels), gut microbiota (bacterial diversity, community structure and membership, and relative abundance of bacterial taxa), and body measurements (body mass, tarsus length, and scaled mass index) across undeveloped, agricultural, and urban areas for medium ground finches (Geospiza fortis) and small ground finches (G. fuliginosa). We found that lysozyme activity was lower and observed bacterial species richness was higher in urban areas compared to non-urban areas across both finch species. For small ground finches, relative abundances of three bacterial genera (Pseudoxanthomonas, Cloacibacterium, and Dietzia spp.) were higher in urban areas compared to non-urban areas, but this pattern was not observed in medium ground finches. Medium ground finches were smaller in undeveloped areas compared to the other two areas, but body measurements of small ground finches did not differ across areas. Our results suggest that human activity can impact immune measures and gut microbiota of Darwins finches.
]]></description>
<dc:creator>Bygrave, J.</dc:creator>
<dc:creator>Zylberberg, M. A.</dc:creator>
<dc:creator>Addesso, A.</dc:creator>
<dc:creator>Knutie, S. A.</dc:creator>
<dc:date>2022-02-09</dc:date>
<dc:identifier>doi:10.1101/2022.02.07.479384</dc:identifier>
<dc:title><![CDATA[Influence of earlier human activity on gut microbiota and immune responses of Darwin's finches in the Galapagos Islands]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.07.479457v1?rss=1">
<title>
<![CDATA[
Deep tissue infection by an invasive human fungal pathogen requires novel lipid-based suppression of the IL-17 response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.07.479457v1?rss=1</link>
<description><![CDATA[
Candida albicans is the most common cause of human fungal infection, but the mechanisms of invasive pathogenesis remain poorly defined. Here we identify an unexpected mechanism: lipid-mediated immunosuppression. Through forward genetics, we found that C. albicans secretes a lipase, Lip2, that is critical for invasive disease. Murine infection with C. albicans strains that lack Lip2 display an exaggerated host IL-17 response that leads to fungal clearance from solid organs and host survival. IL-17 signaling is required for Lip2 action. The lipase activity of Lip2 inhibits IL-17 production indirectly through suppression of IL-23 production by tissue resident dendritic cells. We conclude that C. albicans suppresses antifungal IL-17 defense in solid organs by altering the tissue lipid milieu.
]]></description>
<dc:creator>Basso, P.</dc:creator>
<dc:creator>Dang, E. V.</dc:creator>
<dc:creator>Urisman, A.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:creator>Cowen, L. E.</dc:creator>
<dc:creator>Noble, S. M.</dc:creator>
<dc:date>2022-02-08</dc:date>
<dc:identifier>doi:10.1101/2022.02.07.479457</dc:identifier>
<dc:title><![CDATA[Deep tissue infection by an invasive human fungal pathogen requires novel lipid-based suppression of the IL-17 response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.10.479992v1?rss=1">
<title>
<![CDATA[
Temporal and sequential transcriptional dynamics define lineage shifts in corticogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.10.479992v1?rss=1</link>
<description><![CDATA[
The cerebral cortex contains billions of neurons, and their disorganization or misspecification leads to neurodevelopmental disorders. Understanding how the plethora of projection neuron subtypes are generated by cortical neural stem cells (NSCs) is a major challenge. Here, we focused on elucidating the transcriptional landscape of murine embryonic NSCs, basal progenitors (BPs) and newborn neurons (NBNs) throughout cortical development. We uncover dynamic shifts in transcriptional space over time, and heterogeneity within each progenitor population. We identified signature hallmarks of NSC, BP and NBN clusters, and predict active transcriptional nodes and networks that contribute to neural fate specification. We find that the expression of receptors, ligands and downstream pathway components is highly dynamic over time and throughout the lineage implying differential responsiveness to signals. Thus, we provide an expansive compendium of gene expression during cortical development that will be an invaluable resource for studying neural developmental processes and neurodevelopmental disorders.
]]></description>
<dc:creator>Mukhtar, T.</dc:creator>
<dc:creator>Breda, J.</dc:creator>
<dc:creator>Boareto, M.</dc:creator>
<dc:creator>Grobecker, P.</dc:creator>
<dc:creator>Karimaddini, Z.</dc:creator>
<dc:creator>Grison, A.</dc:creator>
<dc:creator>Eschbach, K.</dc:creator>
<dc:creator>Chandrasekhar, R.</dc:creator>
<dc:creator>Vermeul, S.</dc:creator>
<dc:creator>Okoniewski, M.</dc:creator>
<dc:creator>Pachkov, M.</dc:creator>
<dc:creator>Atanasoski, S.</dc:creator>
<dc:creator>Beisel, C.</dc:creator>
<dc:creator>Iber, D.</dc:creator>
<dc:creator>Nimwegen, E. v.</dc:creator>
<dc:creator>Taylor, V.</dc:creator>
<dc:date>2022-02-11</dc:date>
<dc:identifier>doi:10.1101/2022.02.10.479992</dc:identifier>
<dc:title><![CDATA[Temporal and sequential transcriptional dynamics define lineage shifts in corticogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.15.480601v1?rss=1">
<title>
<![CDATA[
Highly efficient generation of isogenic pluripotent stem cell models using prime editing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.15.480601v1?rss=1</link>
<description><![CDATA[
The recent development of prime editing (PE) genome engineering technologies has the potential to significantly simplify the generation of human pluripotent stem cell (hPSC)-based disease models. PE is a multi-component editing system that uses a Cas9-nickase fused to a reverse transcriptase (nCas9-RT) and an extended PE guide RNA (pegRNA). Once reverse transcribed, the pegRNA extension functions as a repair template to introduce precise designer mutations at the target site. Here, we systematically compared the editing efficiencies of PE to conventional gene editing methods in hPSCs. This analysis revealed that PE is overall more efficient and precise than homology-directed repair (HDR) of site-specific nuclease-induced double-strand breaks (DSBs). Specifically, PE is more effective in generating heterozygous editing events to create autosomal dominant disease-associated mutations. By stably integrating the nCas9-RT into hPSCs we achieved editing efficiencies equal to those reported for cancer cells, suggesting that the expression of the PE components, rather than cell-intrinsic features, limit PE in hPSCs. To improve the efficiency of PE in hPSCs, we optimized the delivery modalities for the PE components. Delivery of the nCas9-RT as mRNA combined with synthetically generated chemically-modified pegRNAs and nicking guide RNAs (ngRNAs) improved editing efficiencies up to 13-fold compared to transfecting the prime editing components as plasmids or ribonucleoprotein particles (RNPs). Finally, we demonstrated that this mRNA-based delivery approach can be used repeatedly to yield editing efficiencies exceeding 60% and to correct or introduce familial mutations causing Parkinsons disease in hPSCs.
]]></description>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Busquets, O.</dc:creator>
<dc:creator>Verma, Y.</dc:creator>
<dc:creator>Syed, K. M.</dc:creator>
<dc:creator>Kutnowski, N.</dc:creator>
<dc:creator>Pangilinan, G. R.</dc:creator>
<dc:creator>Gilbert, L.</dc:creator>
<dc:creator>Bateup, H.</dc:creator>
<dc:creator>Rio, D. C.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:creator>Soldner, F.</dc:creator>
<dc:date>2022-02-15</dc:date>
<dc:identifier>doi:10.1101/2022.02.15.480601</dc:identifier>
<dc:title><![CDATA[Highly efficient generation of isogenic pluripotent stem cell models using prime editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.16.480703v1?rss=1">
<title>
<![CDATA[
Tensor decomposition reveals coordinated multicellular patterns of transcriptional variation that distinguish and stratify disease individuals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.16.480703v1?rss=1</link>
<description><![CDATA[
Tissue- and organism-level biological processes often involve coordinated action of multiple distinct cell types. Current computational methods for the analysis of single-cell RNA-sequencing (scRNA-seq) data, however, are not designed to capture co-variation of cell states across samples, in part due to the low number of biological samples in most scRNA-seq datasets. Recent advances in sample multiplexing have enabled population-scale scRNA-seq measurements of tens to hundreds of samples. To take advantage of such datasets, here we introduce a computational approach called single-cell Interpretable Tensor Decomposition (scITD). This method extracts "multicellular gene expression patterns" that capture how sample-specific expression states of a cell type are correlated with the expression states of other cell types. Such multicellular patterns can reveal molecular mechanisms underlying coordinated changes of different cell types within the tissue, and can be used to stratify individuals in a clinically-relevant and reproducible manner. We first validated the performance of scITD using in vitro experimental data and simulations. We then applied scITD to scRNA-seq data on peripheral blood mononuclear cells (PBMCs) from 115 patients with systemic lupus erythematosus and 56 healthy controls. We recapitulated a well-established pan-cell-type signature of interferon-signaling that was associated with the presence of anti-dsDNA autoantibodies and a disease activity index. We further identified a novel multicellular pattern linked to nephritis, which was characterized by an expansion of activated memory B cells along with helper T cell activation. Our approach also sheds light on ligand-receptor interactions potentially mediating these multicellular patterns. As validation, we demonstrated that these expression patterns also stratified donors from a pediatric SLE dataset by the same phenotypic attributes. Lastly, we found the interferon multicellular pattern and others to be conserved in a COVID-19 dataset, pointing to the presence of both general and disease-specific patterns of inter-individual immune variation. Overall, scITD is a flexible method for exploring co-variation of cell states in multi-sample single-cell datasets, which can yield new insights into complex non-cell-autonomous dependencies that define and stratify disease.
]]></description>
<dc:creator>Mitchel, J.</dc:creator>
<dc:creator>Gordon, M. G.</dc:creator>
<dc:creator>Perez, R. K.</dc:creator>
<dc:creator>Biederstedt, E.</dc:creator>
<dc:creator>Bueno, R.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Kharchenko, P.</dc:creator>
<dc:date>2022-02-18</dc:date>
<dc:identifier>doi:10.1101/2022.02.16.480703</dc:identifier>
<dc:title><![CDATA[Tensor decomposition reveals coordinated multicellular patterns of transcriptional variation that distinguish and stratify disease individuals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.16.480729v1?rss=1">
<title>
<![CDATA[
Antigen-dependent inducible T cell reporter system for PET imaging of breast cancer and glioblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.16.480729v1?rss=1</link>
<description><![CDATA[
For the past several decades, chimeric antigen receptor T cell (CAR T) therapies have shown promise in the treatment of cancers. These treatments would greatly benefit from companion imaging biomarkers to follow the trafficking of T cells in vivo. Using synthetic biology, we engineered T cells with a chimeric receptor SyNthetic Intramembrane Proteolysis Receptor (SNIPR) that induces overexpression of an exogenous reporter gene cassette upon recognition of specific tumor markers. We then applied a SNIPR-based positron emission tomography (PET) reporter system to two cancer-relevant antigens, human epidermal growth factor receptor 2 (HER2) and epidermal growth factor receptor variant III (EGFRvIII), commonly expressed in breast and glial tumors respectively. Antigen-specific reporter induction of the SNIPR-PET T cells was confirmed in vitro using GFP fluorescence, luciferase luminescence, and the HSV-TK PET reporter with [18F]FHBG. T cells associated with their target antigens were successfully imaged using PET in dual xenograft HER2+/HER2- and EGFRvIII+/EGFRvIII-animal models, with > 10-fold higher [18F]FHBG signals seen in antigen-expressing tumors versus the corresponding controls. The main innovation described is therefore PET detection of T cells via specific antigen-induced signals, in contrast to reporter systems relying on constitutive gene expression.
]]></description>
<dc:creator>Shin, J.</dc:creator>
<dc:creator>Parker, M.</dc:creator>
<dc:creator>Zhu, I.</dc:creator>
<dc:creator>Alanizi, A.</dc:creator>
<dc:creator>Rodriguez, C.</dc:creator>
<dc:creator>Liu, R.</dc:creator>
<dc:creator>Watchmaker, P.</dc:creator>
<dc:creator>Kalita, M.</dc:creator>
<dc:creator>Blecha, J.</dc:creator>
<dc:creator>Luu, J.</dc:creator>
<dc:creator>Wright, B.</dc:creator>
<dc:creator>Lapi, S.</dc:creator>
<dc:creator>Flavell, R.</dc:creator>
<dc:creator>Okada, H.</dc:creator>
<dc:creator>Tlsty, T.</dc:creator>
<dc:creator>Wilson, D.</dc:creator>
<dc:creator>Roybal, K.</dc:creator>
<dc:date>2022-02-16</dc:date>
<dc:identifier>doi:10.1101/2022.02.16.480729</dc:identifier>
<dc:title><![CDATA[Antigen-dependent inducible T cell reporter system for PET imaging of breast cancer and glioblastoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.16.480779v1?rss=1">
<title>
<![CDATA[
Inhibition of the tuft cell/ILC2 axis reduces gastric tumor development in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.16.480779v1?rss=1</link>
<description><![CDATA[
Although gastric cancer is a leading cause of cancer-related deaths, systemic treatment strategies remain scarce. Here we explore a metabolite-triggered circuit between epithelial tuft cells and innate lymphoid type 2 cells (ILC2) that is evolutionarily optimized for intestinal remodeling in response to helminth infection. We demonstrate that tuft cell-derived interleukin 25 (IL25) acts as an alarmin on ILC2s to induce the release of IL13 as a growth factor for tuft cells, and propose that this model drives early metaplastic remodeling and gastric tumor formation. Genetic ablation of tuft cells, ILC2s or antibody-mediated neutralization of IL13 or IL25 reduces the growth of established tumors. Thus, the tuft cell/ILC2 axis provides an opportunity to therapeutically inhibit preneoplastic lesions and early-stage gastric cancer through repurposing of antibody-mediated therapies.

One-Sentence SummaryTuft cells and type 2 innate lymphoid cells offer a new therapeutic target in gastric disease.
]]></description>
<dc:creator>O'Keefe, R. N.</dc:creator>
<dc:creator>Carli, A.</dc:creator>
<dc:creator>Baloyan, D.</dc:creator>
<dc:creator>Sterle, S.</dc:creator>
<dc:creator>Eissmann, M. F.</dc:creator>
<dc:creator>Poh, A. R.</dc:creator>
<dc:creator>Seillet, C.</dc:creator>
<dc:creator>Locksley, R. M.</dc:creator>
<dc:creator>Ernst, M. M.</dc:creator>
<dc:creator>Buchert, M.</dc:creator>
<dc:date>2022-02-18</dc:date>
<dc:identifier>doi:10.1101/2022.02.16.480779</dc:identifier>
<dc:title><![CDATA[Inhibition of the tuft cell/ILC2 axis reduces gastric tumor development in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.17.480942v1?rss=1">
<title>
<![CDATA[
Single-Cell Multi-Omic Roadmap of Human Fetal Pancreatic Development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.17.480942v1?rss=1</link>
<description><![CDATA[
The critical cellular transitions that govern human pancreas development are largely unknown. We performed large-scale single-cell RNA-sequencing (scRNA-Seq) to interrogate human fetal pancreas development from 8-20 weeks post conception. We identified 103 distinct cell types, including four novel endocrine progenitor subtypes displaying unique transcriptional features and differentiation potency. Integration with single-nucleus Assay for Transposase Accessible Chromatin Sequencing (snATAC-Seq) identified candidate regulators of human endocrine cell fate and revealed development-specific regulatory annotation at diabetes risk loci. Comparison of in vitro stem cell-derived and endogenous endocrine cells predicted aberrant genetic programs leading to the generation of off-target cells. Finally, knock-out studies revealed that the gene FEV regulates human endocrine differentiation. This work establishes a roadmap of human pancreatic development, highlights previously unappreciated cellular diversity and lineage dynamics, and provides a blueprint for understanding pancreatic disease and physiology, as well as generating human stem cell-derived islet cells in vitro for regenerative medicine purposes.
]]></description>
<dc:creator>de la O, S.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Sun, H.</dc:creator>
<dc:creator>Yu, S. K.</dc:creator>
<dc:creator>Wong, D. M.</dc:creator>
<dc:creator>Chu, E.</dc:creator>
<dc:creator>Rao, S. A.</dc:creator>
<dc:creator>Eng, N.</dc:creator>
<dc:creator>Peixoto, G.</dc:creator>
<dc:creator>Bouza, J.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Knox, S. M.</dc:creator>
<dc:creator>Tward, A. D.</dc:creator>
<dc:creator>Gloyn, A. L.</dc:creator>
<dc:creator>Sneddon, J. B.</dc:creator>
<dc:date>2022-02-18</dc:date>
<dc:identifier>doi:10.1101/2022.02.17.480942</dc:identifier>
<dc:title><![CDATA[Single-Cell Multi-Omic Roadmap of Human Fetal Pancreatic Development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.19.481149v1?rss=1">
<title>
<![CDATA[
Statistical curve models for inferring 3D chromatin architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.19.481149v1?rss=1</link>
<description><![CDATA[
Reconstructing three dimensional (3D) chromatin structure from conformation capture assays (such as Hi-C) is a critical task in computational biology, since chromatin spatial architecture plays a vital role in numerous cellular processes and direct imaging is challenging. We previously introduced Poisson metric scaling (PoisMS), a technique that models chromatin by a smooth curve, which yielded promising results. In this paper, we advance several ways for improving PoisMS. In particular, we address initialization issues by using a smoothing spline basis. The resulting SPoisMS method produces a sequence of reconstructions re-using previous solutions as warm starts. Importantly, this approach permits smoothing degree to be determined via cross-validation which was problematic using our prior B-spline basis. In addition, motivated by the sparsity of Hi-C contact data, especially when obtained from single-cell assays, we appreciably extend the class of distributions used to model contact counts. We build a general distribution-based metric scaling (DBMS) framework, from which we develop zero-inflated and Hurdle Poisson models as well as negative binomial applications. Illustrative applications make recourse to bulk Hi-C data from IMR90 cells and single-cell Hi-C data from mouse embryonic stem cells.
]]></description>
<dc:creator>Tuzhilina, E.</dc:creator>
<dc:creator>Hastie, T.</dc:creator>
<dc:creator>Segal, M.</dc:creator>
<dc:date>2022-02-20</dc:date>
<dc:identifier>doi:10.1101/2022.02.19.481149</dc:identifier>
<dc:title><![CDATA[Statistical curve models for inferring 3D chromatin architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.21.481343v1?rss=1">
<title>
<![CDATA[
Aberrant cortical development is driven by impaired cell cycle and translational control in a DDX3X syndrome model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.21.481343v1?rss=1</link>
<description><![CDATA[
Mutations in the RNA helicase, DDX3X, are a leading cause of Intellectual Disability and present as DDX3X syndrome, a neurodevelopmental disorder associated with cortical malformations and autism. Yet the cellular and molecular mechanisms by which DDX3X controls cortical development are largely unknown. Here, using a mouse model of Ddx3x loss-of-function we demonstrate that DDX3X directs translational and cell cycle control of neural progenitors, which underlies precise corticogenesis. First, we show brain development is highly sensitive to Ddx3x dosage; Complete Ddx3x loss from neural progenitors causes microcephaly in females, whereas hemizygous males and heterozygous females show reduced neurogenesis without marked microcephaly. In addition, Ddx3x loss is sexually dimorphic, as its paralog, Ddx3y, compensates for Ddx3x in the developing male neocortex. Using live imaging of progenitors, we show that DDX3X promotes neuronal generation by regulating both cell cycle duration and neurogenic divisions. Finally, we use ribosome profiling in vivo to discover the repertoire of translated transcripts in neural progenitors, including those which are DDX3X-dependent and essential for neurogenesis. Our study reveals invaluable new insights into the etiology of DDX3X syndrome, implicating dysregulated progenitor cell cycle dynamics and translation as pathogenic mechanisms.
]]></description>
<dc:creator>Hoye, M. L.</dc:creator>
<dc:creator>Calviello, L.</dc:creator>
<dc:creator>Poff, A. J.</dc:creator>
<dc:creator>Ejimogu, N.-E.</dc:creator>
<dc:creator>Newman, C. R.</dc:creator>
<dc:creator>ou, j.</dc:creator>
<dc:creator>Floor, S. N.</dc:creator>
<dc:creator>Silver, D. L.</dc:creator>
<dc:date>2022-02-21</dc:date>
<dc:identifier>doi:10.1101/2022.02.21.481343</dc:identifier>
<dc:title><![CDATA[Aberrant cortical development is driven by impaired cell cycle and translational control in a DDX3X syndrome model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.23.481517v1?rss=1">
<title>
<![CDATA[
T cells use distinct topological and membrane receptor scanning strategies that individually coalesce during receptor recognition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.23.481517v1?rss=1</link>
<description><![CDATA[
During immune surveillance, CD8 T cells scan the surface of antigen presenting cells using dynamic microvillar palpation and movements as well as by having their receptors pre-concentrated into patches. Here, we use real-time lattice light sheet microscopy to demonstrate the independence of microvillar and membrane receptor patch scanning. While T cell receptor (TCR) patches can distribute to microvilli, they do so stochastically and not preferentially as for other receptors such as CD62L. The distinctness of TCR patch movement from microvillar movement extends to many other receptors that form patches that also scan independently of the TCR. An exception to this is the CD8 co-receptor which largely co-migrates in patches that overlap with or are closely adjacent to those containing TCRs. Microvilli that assemble into a synapse contain various arrays of the engaged patches, notably of TCRs and the inhibitory receptor PD-1, creating a pastiche of occupancies that vary from microvillar contact to contact. In summary, this work demonstrates that localization of receptor patches within the membrane and on microvillar projections is stochastic prior to antigen detection and that such stochastic variation may play into the generation of many individually-composed receptor patch compositions at a single synapse.

Significance statementMotile T cell microvilli palpate surfaces to facilitate surface scanning in a pattern that is independent of the movement of pre-formed patches of transmembrane antigen-receptors across those microvilli; once T cell receptors engage, the microvilli act to scaffold multiple receptors within a microvillar close-contact.
]]></description>
<dc:creator>Cai, E.</dc:creator>
<dc:creator>Beppler, C.</dc:creator>
<dc:creator>Eichorst, J.</dc:creator>
<dc:creator>Marchuk, K.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2022-02-25</dc:date>
<dc:identifier>doi:10.1101/2022.02.23.481517</dc:identifier>
<dc:title><![CDATA[T cells use distinct topological and membrane receptor scanning strategies that individually coalesce during receptor recognition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.28.482366v1?rss=1">
<title>
<![CDATA[
"Transposon Mutagenesis Reveals RBMS3 as a Promoter of Malignant Progression of BRAFV600E-Driven Lung Tumorigenesis." 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.28.482366v1?rss=1</link>
<description><![CDATA[
Mutationally-activated BRAFV600E is detected in ~2% of all human non-small cell lung cancers (NSCLC), and serves as a predictive biomarker for treatment of patients with FDA-approved pathway-targeted therapies that inhibit signaling by the BRAFV600E oncoprotein kinase. In genetically engineered mouse (GEM) models, expression of BRAFV600E in alveolar type 2 (AT2) pneumocytes initiates the development of benign lung tumors that, without additional genetic alterations, rarely progress to malignant lung adenocarcinomas. To identify genes that might cooperate with BRAFV600E for malignant lung cancer progression we employed Sleeping Beauty (SB)-mediated transposon mutagenesis, which dramatically accelerated the onset of lethal lung adenocarcinomas. Amongst the diverse group of genes identified by this in vivo screen was Rbms3 (RNA binding motif single-stranded interacting protein 3), an RNA-binding protein implicated as a possible tumor suppressor. Using CRISPR/CAS9 gene editing we confirmed that RBMS3 silencing cooperated with BRAFV600E to promote progression of malignant lung cancer with a distinct micropapillary architecture. Moreover, RBMS3 silencing also cooperated with BRAFV600E to promote the growth of lung organoids in vitro. BRAFV600E/RBMS3Null lung tumors displayed elevated expression of b-catenin (CTNNB1), suggesting that RBMS3 silencing may result in elevated signaling through the WNT>CTNNB1>c-MYC pathway. Finally, analyses of patient samples in The Cancer Genome Atlas (TCGA) revealed that the region of chromosome 3 encompassing RBMS3 is frequently lost in NSCLC and correlates with poor patient prognosis. Collectively, SB-mediated transposon mutagenesis has revealed the ability of a novel tumor suppressor, RBMS3, to cooperate with BRAFV600E to promote lung carcinogenesis, and suggests that RBMS3 silencing may contribute to malignant progression of numerous human lung cancers.

SIGNIFICANCEThe BRAFV600E oncoprotein kinase is a potent initiator of benign lung tumorigenesis, but is insufficient to elicit malignant lung adenocarcinoma without additional cooperating alterations. Sleeping Beauty-mediated transposon mutagenesis has revealed a number of genes that cooperate with BRAFV600E to promote lung cancer progression, in particular Rbms3, which encodes an RNA binding protein. Hence, this genetic screen provides a deeper understanding of the molecular mechanisms underlying BRAFV600E-driven lung carcinogenesis, and is an important step improving our ability to successfully target this disease.
]]></description>
<dc:creator>Vaishnavi, A.</dc:creator>
<dc:creator>Juan, J.</dc:creator>
<dc:creator>Scherzer, M. T.</dc:creator>
<dc:creator>VanVeen, J. E.</dc:creator>
<dc:creator>Stehn, C.</dc:creator>
<dc:creator>Hackett, C. S.</dc:creator>
<dc:creator>Dupuy, A.</dc:creator>
<dc:creator>Chmura, S. A.</dc:creator>
<dc:creator>van der Weyden, L.</dc:creator>
<dc:creator>Newberg, J. Y.</dc:creator>
<dc:creator>Mann, K. M.</dc:creator>
<dc:creator>Liu, A.</dc:creator>
<dc:creator>Rust, A. G.</dc:creator>
<dc:creator>Weiss, W. A.</dc:creator>
<dc:creator>Adams, D.</dc:creator>
<dc:creator>Grossman, A.</dc:creator>
<dc:creator>Mann, M. B.</dc:creator>
<dc:creator>McMahon, M.</dc:creator>
<dc:date>2022-02-28</dc:date>
<dc:identifier>doi:10.1101/2022.02.28.482366</dc:identifier>
<dc:title><![CDATA["Transposon Mutagenesis Reveals RBMS3 as a Promoter of Malignant Progression of BRAFV600E-Driven Lung Tumorigenesis."]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.01.482549v1?rss=1">
<title>
<![CDATA[
Sub-harmonic Entrainment of Cortical Gamma Oscillations to Deep Brain Stimulation in Parkinson's Disease: Predictions and Validation of a Patient-Specific Non-Linear Model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.01.482549v1?rss=1</link>
<description><![CDATA[
ObjectivesThe exact mechanisms of deep brain stimulation (DBS) are still an active area of investigation, in spite of its clinical successes. This is due in part to the lack of understanding of the effects of stimulation on neuronal rhythms. Entrainment of brain oscillations has been hypothesised as a potential mechanism of neuromodulation. A better understanding of entrainment might further inform existing methods of continuous DBS, and help refine algorithms for adaptive methods. The purpose of this study is to develop and test a theoretical framework to predict entrainment of cortical rhythms to DBS across a wide range of stimulation parameters.

Materials and MethodsWe fit a model of interacting neural populations to selected features characterising PD patients off-stimulation finely-tuned gamma rhythm recorded through electrocorticography. Using the fitted models, we predict basal ganglia DBS parameters that would result in 1:2 entrainment, a special case of sub-harmonic entrainment observed in patients and predicted by theory.

ResultsWe show that the neural circuit models fitted to patient data exhibit 1:2 entrainment when stimulation is provided across a range of stimulation parameters. Furthermore, we verify key features of the region of 1:2 entrainment in the stimulation frequency/amplitude space with follow-up recordings from the same patients, such as the loss of 1:2 entrainment above certain stimulation amplitudes.

ConclusionOur results reveal that continuous, constant frequency DBS in patients may lead to nonlinear patterns of neuronal entrainment across stimulation parameters, and that these responses can be predicted by modelling. Should entrainment prove to be an important mechanism of therapeutic stimulation, our modelling framework may reduce the parameter space that clinicians must consider when programming devices for optimal benefit.
]]></description>
<dc:creator>Sermon, J. J.</dc:creator>
<dc:creator>Olaru, M.</dc:creator>
<dc:creator>Anso, J.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:creator>Bogacz, R.</dc:creator>
<dc:creator>Starr, P. A.</dc:creator>
<dc:creator>Denison, T.</dc:creator>
<dc:creator>Duchet, B.</dc:creator>
<dc:date>2022-03-04</dc:date>
<dc:identifier>doi:10.1101/2022.03.01.482549</dc:identifier>
<dc:title><![CDATA[Sub-harmonic Entrainment of Cortical Gamma Oscillations to Deep Brain Stimulation in Parkinson's Disease: Predictions and Validation of a Patient-Specific Non-Linear Model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.06.483172v1?rss=1">
<title>
<![CDATA[
Low expression of EXOSC2 protects against clinical COVID-19 and impedes SARS-CoV-2 replication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.06.483172v1?rss=1</link>
<description><![CDATA[
New therapeutic targets are a valuable resource in the struggle to reduce the morbidity and mortality associated with the COVID-19 pandemic, caused by the SARS-CoV-2 virus. Genome-wide association studies (GWAS) have identified risk loci, but some loci are associated with co-morbidities and are not specific to host-virus interactions. Here, we identify and experimentally validate a link between reduced expression of EXOSC2 and reduced SARS-CoV-2 replication. EXOSC2 was one of 332 host proteins examined, all of which interact directly with SARS-CoV-2 proteins; EXOSC2 interacts with Nsp8 which forms part of the viral RNA polymerase. Lung-specific eQTLs were identified from GTEx (v7) for each of the 332 host proteins. Aggregating COVID-19 GWAS statistics for gene-specific eQTLs revealed an association between increased expression of EXOSC2 and higher risk of clinical COVID-19 which survived stringent multiple testing correction. EXOSC2 is a component of the RNA exosome and indeed, LC-MS/MS analysis of protein pulldowns demonstrated an interaction between the SARS-CoV-2 RNA polymerase and the majority of human RNA exosome components. CRISPR/Cas9 introduction of nonsense mutations within EXOSC2 in Calu-3 cells reduced EXOSC2 protein expression, impeded SARS-CoV-2 replication and upregulated oligoadenylate synthase (OAS) genes, which have been linked to a successful immune response against SARS-CoV-2. Reduced EXOSC2 expression did not reduce cellular viability. OAS gene expression changes occurred independent of infection and in the absence of significant upregulation of other interferon-stimulated genes (ISGs). Targeted depletion or functional inhibition of EXOSC2 may be a safe and effective strategy to protect at-risk individuals against clinical COVID-19.
]]></description>
<dc:creator>Moll, T.</dc:creator>
<dc:creator>Odon, V.</dc:creator>
<dc:creator>Harvey, C.</dc:creator>
<dc:creator>Collins, M.</dc:creator>
<dc:creator>Peden, A.</dc:creator>
<dc:creator>Franklin, J.</dc:creator>
<dc:creator>Graves, E.</dc:creator>
<dc:creator>Marshall, J.</dc:creator>
<dc:creator>dos Santos Souza, C.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Azzouz, M.</dc:creator>
<dc:creator>Gordon, D.</dc:creator>
<dc:creator>Krogan, N.</dc:creator>
<dc:creator>Ferraiuolo, L.</dc:creator>
<dc:creator>Snyder, M.</dc:creator>
<dc:creator>Shaw, P.</dc:creator>
<dc:creator>Rehwinkel, J.</dc:creator>
<dc:creator>Cooper-Knock, J.</dc:creator>
<dc:date>2022-03-07</dc:date>
<dc:identifier>doi:10.1101/2022.03.06.483172</dc:identifier>
<dc:title><![CDATA[Low expression of EXOSC2 protects against clinical COVID-19 and impedes SARS-CoV-2 replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.07.483367v1?rss=1">
<title>
<![CDATA[
Demuxafy: Improvement in droplet assignment by integrating multiple single-cell demultiplexing and doublet detection methods 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.07.483367v1?rss=1</link>
<description><![CDATA[
Recent innovations in droplet-based single-cell RNA-sequencing (scRNA-seq) have provided the technology necessary to investigate biological questions at cellular resolution. With the ability to assay thousands of cells in a single capture, pooling cells from multiple individuals has become a common strategy. Droplets can subsequently be assigned to a specific individual by leveraging their inherent genetic differences, and numerous computational methods have been developed to address this problem. However, another challenge implicit with droplet-based scRNA-seq is the occurrence of doublets - droplets containing two or more cells. The inaccurate assignment of cells to individuals or failure to remove doublets contribute unwanted noise to the data and result in erroneous scientific conclusions. Therefore, it is essential to assign cells to individuals and remove doublets accurately. We present a new framework to improve individual singlet classification and doublet removal through a multi-method intersectional approach.

We developed a framework to evaluate the enhancement in donor assignment and doublet removal through the consensus intersection of multiple demultiplexing and doublet detecting methods. The accuracy was assessed using scRNA-seq data of [~]1.4 million peripheral blood mononucleated cells from 1,034 unrelated individuals and [~]90,000 fibroblast cells from 81 unrelated individuals. We show that our approach significantly improves droplet assignment by separating singlets from doublets and classifying the correct individual compared to any single method. We show that the best combination of techniques varies under different biological and experimental conditions, and we present a framework to optimise cell assignment for a given experiment. We offer Demuxafy (https://demultiplexing-doublet-detecting-docs.readthedocs.io/en/latest/index.html) - a framework built-in Singularity to provide clear, consistent documentation of each method and additional tools to simplify and improve demultiplexing and doublet removal. Our results indicate that leveraging multiple demultiplexing and doublet detecting methods improves accuracy and, consequently, downstream analyses in multiplexed scRNA-seq experiments.
]]></description>
<dc:creator>Neavin, D.</dc:creator>
<dc:creator>Senabouth, A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Ripoll, A.</dc:creator>
<dc:creator>Franke, L.</dc:creator>
<dc:creator>Prabhakar, S.</dc:creator>
<dc:creator>Ye, C.</dc:creator>
<dc:creator>McCarthy, D.</dc:creator>
<dc:creator>Mele, M.</dc:creator>
<dc:creator>Hemberg, M.</dc:creator>
<dc:creator>Powell, J.</dc:creator>
<dc:date>2022-03-08</dc:date>
<dc:identifier>doi:10.1101/2022.03.07.483367</dc:identifier>
<dc:title><![CDATA[Demuxafy: Improvement in droplet assignment by integrating multiple single-cell demultiplexing and doublet detection methods]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.09.483562v1?rss=1">
<title>
<![CDATA[
Whole brain network analysis of neural synchrony and information flow during transition from wakefulness to light non-rapid eye movement sleep 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.09.483562v1?rss=1</link>
<description><![CDATA[
Sleep is a highly stereotyped phenomenon, requiring robust spatial and temporal coordination of neural activity. How the brain coordinates neural activity with sleep onset can provide insight into the physiological functions subserved by sleep and pathologic phenomena associated with sleep onset. We quantified whole-brain network changes in synchrony and information flow during the transition from wake to non-rapid eye movement (NREM) sleep using magnetoencephalography imaging in healthy subjects. In addition, we performed computational modeling to infer excitatory and inhibitory properties of local neural activity. The sleep transition was identified to be encoded in spatially and temporally specific patterns of local and long-range neural synchrony. Patterns of information flow revealed that mesial frontal regions receive hierarchically organized inputs from broad cortical regions upon sleep onset. Finally, biophysical neural mass modeling demonstrated spatially heterogeneous properties of cortical excitation-to-inhibition from wake to NREM. Together, these findings reveal whole-brain corticocortical structure in the sleep-wake transition and demonstrate the orchestration of local and long-range, frequency-specific cortical interactions that are fundamental to sleep onset.
]]></description>
<dc:creator>Fan, J. M.</dc:creator>
<dc:creator>Kudo, K.</dc:creator>
<dc:creator>Verma, P.</dc:creator>
<dc:creator>Ranasinghe, K. G.</dc:creator>
<dc:creator>Morise, H.</dc:creator>
<dc:creator>Findlay, A. M.</dc:creator>
<dc:creator>Kirsch, H. E.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Krystal, A. D.</dc:creator>
<dc:creator>Nagarajan, S. S.</dc:creator>
<dc:date>2022-03-11</dc:date>
<dc:identifier>doi:10.1101/2022.03.09.483562</dc:identifier>
<dc:title><![CDATA[Whole brain network analysis of neural synchrony and information flow during transition from wakefulness to light non-rapid eye movement sleep]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.09.483674v1?rss=1">
<title>
<![CDATA[
Cohesin erases genomic-proximity biases to drive stochastic Protocadherin expression for proper neural wiring 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.09.483674v1?rss=1</link>
<description><![CDATA[
Clustered Protocadherin (Pcdh) proteins act as cell-surface recognition barcodes for neural circuit formation. Neurites expressing the same barcode repel each other, but this mechanism is deployed in two different ways. For instance, convergence of olfactory sensory neuron (OSN) projections requires stochastic expression of distinct Pcdh isoforms in individual cells, while tiling of neural arbors of serotonergic neurons (5-HTs) requires expression of the same isoform, Pcdhc2. Despite their essential role, however, the molecular mechanisms of cell-type specific Pcdh barcoding remain a mystery. Here, we uncover a new role of cohesin: that of regulating distance-independent enhancer-promoter interactions to enable random Pcdh isoform choice via DNA loop extrusion in OSNs. Remarkably, this step mediates DNA demethylation of Pcdh promoters and their CTCF binding sites, thus directing CTCF to the chosen promoter. In contrast, the uniform pattern of Pcdh expression in 5-HTs is achieved through conventional cohesin-independent, distance-dependent enhancer/promoter interactions, that favor choice of the nearest isoform. Thus, cell-type specific cohesin deployment converts a distance-dependent and deterministic regulatory logic into a distance-independent and stochastic one. We propose that this mechanism provides an elegant strategy to achieve distinct patterns of Pcdh expression that generate wiring instructions to meet the connectivity requirements of different neural classes.
]]></description>
<dc:creator>Kiefer, L.</dc:creator>
<dc:creator>Servito, G. I. F.</dc:creator>
<dc:creator>Rajkumar, S. M.</dc:creator>
<dc:creator>Langen, J.</dc:creator>
<dc:creator>Chiosso, A.</dc:creator>
<dc:creator>Buckley, A.</dc:creator>
<dc:creator>Cha, E. S.</dc:creator>
<dc:creator>Horta, A.</dc:creator>
<dc:creator>Mui, M. H.</dc:creator>
<dc:creator>Canzio, D.</dc:creator>
<dc:date>2022-03-11</dc:date>
<dc:identifier>doi:10.1101/2022.03.09.483674</dc:identifier>
<dc:title><![CDATA[Cohesin erases genomic-proximity biases to drive stochastic Protocadherin expression for proper neural wiring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.09.483691v1?rss=1">
<title>
<![CDATA[
The maternal X chromosome impairs cognition and accelerates brain aging through epigenetic modulation in female mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.09.483691v1?rss=1</link>
<description><![CDATA[
Female mammalian cells harbor two X chromosomes, one of maternal and one of paternal origin. During development, one X randomly inactivates1-4. This renders either the maternal or paternal X active, causing X mosaicism that varies among individual females, with some showing considerable or complete skew in the general female population5-7. Parent-of-X-origin can modify epigenetics via DNA methylation8, 9 and possibly gene expression; thus, mosaicism could buffer dysregulated processes in aging and disease. However, whether X skewing - or its mosaicism - alters functions in females is largely unknown. Here we tested whether skew toward the maternal X (Xm) influences key functions of the body. Among cardiac, bone, metabolic, and brain functions, Xm selectively impaired cognition in female mice throughout the lifespan. Cognitive deficits were accompanied by Xm-mediated acceleration of biologic or epigenetic aging of the female hippocampus, a key center for learning and memory. Xm showed epigenetic imprinting of several genes within hippocampal neurons, suggesting silenced cognitive loci. Thus, the maternal X chromosome impaired cognition, accelerated brain aging, and silenced genes. Understanding how the maternal X impairs brain function could lead to new understanding of female heterogeneity in cognitive heath and to X chromosome-derived pathways against cognitive deficits and brain aging.
]]></description>
<dc:creator>Abdulai-Saiku, S.</dc:creator>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Moreno, A.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Panning, B.</dc:creator>
<dc:creator>Dubal, D.</dc:creator>
<dc:date>2022-03-12</dc:date>
<dc:identifier>doi:10.1101/2022.03.09.483691</dc:identifier>
<dc:title><![CDATA[The maternal X chromosome impairs cognition and accelerates brain aging through epigenetic modulation in female mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.10.483861v1?rss=1">
<title>
<![CDATA[
Thermodynamic stabilization of the von Willebrand Factor A1 domain due to loss-of-function disease-related mutations. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.10.483861v1?rss=1</link>
<description><![CDATA[
The von Willebrand disease (vWD) is the most common hereditary bleeding disorder, caused by defects of the von Willebrand Factor (vWF), a large extracellular protein in charge of adhering platelets at sites of vascular lesion. vWF carries out this essential homeostatic task, via the specific protein-protein interaction between the vWF A1 domain and the platelet receptor, the glycoprotein Ib alpha (GPIB). Upon the vWF activation triggered by the shear of the flowing blood. The two naturally occurring mutations G1324A and G1324S at the A1 domain, near the GPIB binding site, result in a dramatic decrease of platelets adhesion, a bleeding disorder classified as type 2M vWD. However, it remained unclear how these two supposedly minor modifications lead to this drastic phenotypic response. We addressed this question using a combination of equilibrium-molecular dynamics (MD) and non-equilibrium MD-based free energy simulations. Our data confirm that both mutations maintain the highly stable Rossmann fold of the vWF A1 domain. These mutations locally diminished the flexibility of the binding site to GPIB and induced a conformational change that affected the nearby secondary structure elements. Furthermore, we observed two significant changes in the vWF A1 domain upon mutation, the global redistribution of the internal mechanical stress and the increased thermodynamic stability of the A1 domain. These observations are consistent with previously-reported mutation-augmented melting temperatures. Overall, our results support the idea of thermodynamic conformational restriction of A1-- before the binding to GPIB--as a crucial factor determining the loss-of-function of the G1324A(S) vWD mutants.
]]></description>
<dc:creator>Sandoval-Perez, A.</dc:creator>
<dc:creator>Mejia-Restrepo, V.</dc:creator>
<dc:creator>Aponte-Santamaria, C.</dc:creator>
<dc:date>2022-03-11</dc:date>
<dc:identifier>doi:10.1101/2022.03.10.483861</dc:identifier>
<dc:title><![CDATA[Thermodynamic stabilization of the von Willebrand Factor A1 domain due to loss-of-function disease-related mutations.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.15.484467v1?rss=1">
<title>
<![CDATA[
A conserved immune trajectory of recovery in hospitalized COVID-19 patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.15.484467v1?rss=1</link>
<description><![CDATA[
Many studies have provided insights into the immune response to COVID-19; however, little is known about the immunological changes and immune signaling occurring during COVID-19 resolution. Individual heterogeneity and variable disease resolution timelines obscure unifying immune characteristics. Here, we collected and profiled >200 longitudinal peripheral blood samples from patients hospitalized with COVID-19, with other respiratory infections, and healthy individuals, using mass cytometry to measure immune cells and signaling states at single cell resolution. COVID-19 patients showed a unique immune composition and an early, coordinated and elevated immune cell signaling profile, which correlated with early hospital discharge. Intra-patient time course analysis tied to clinically relevant events of recovery revealed a conserved set of immunological processes that accompany, and are unique to, disease resolution and discharge. This immunological process, together with additional changes in CD4 regulatory T cells and basophils, accompanies recovery from respiratory failure and is associated with better clinical outcomes at the time of admission. Our work elucidates the biological timeline of immune recovery from COVID-19 and provides insights into the fundamental processes of COVID-19 resolution in hospitalized patients.
]]></description>
<dc:creator>Okholm, T. L. H.</dc:creator>
<dc:creator>Burnett, C. E.</dc:creator>
<dc:creator>Tenvooren, I.</dc:creator>
<dc:creator>Marquez, D. M.</dc:creator>
<dc:creator>Tamaki, S.</dc:creator>
<dc:creator>Sandoval, P. M.</dc:creator>
<dc:creator>The UCSF COMET Consortium,</dc:creator>
<dc:creator>Calfee, C. S.</dc:creator>
<dc:creator>Hendrickson, C. M.</dc:creator>
<dc:creator>Kangelaris, K. N.</dc:creator>
<dc:creator>Langelier, C. R.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Woodruff, P. G.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:creator>Ansel, K. M.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:date>2022-03-16</dc:date>
<dc:identifier>doi:10.1101/2022.03.15.484467</dc:identifier>
<dc:title><![CDATA[A conserved immune trajectory of recovery in hospitalized COVID-19 patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.21.485195v1?rss=1">
<title>
<![CDATA[
Regionally distinct trophoblast regulate barrier function and invasion in the human placenta 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.21.485195v1?rss=1</link>
<description><![CDATA[
The human placenta contains two specialized regions: the villous chorion where gases and nutrients are exchanged between maternal and fetal blood, and the smooth chorion which surrounds more than 70% of the developing fetus but whose cellular composition and function is poorly understood. Here, we use single cell RNA sequencing to compare the cell types and molecular programs between these two regions in the second trimester human placenta. Each region consists of progenitor cytotrophoblasts (CTBs) and extravillous trophoblasts (EVTs) with similar gene expression programs. While CTBs in the villous chorion differentiate into syncytiotrophoblasts, they take an alternative trajectory in the smooth chorion producing a previously unknown CTB population which we term smooth-chorion-specific CTBs (SC-CTBs). Marked by expression of region-specific cytokeratins, the SC-CTBs form a stratified epithelium above a basal layer of progenitor CTBs. They express epidermal and metabolic transcriptional programs consistent with a primary role in defense against physical stress and pathogens. Additionally, we show that SC-CTBs closely associate with EVTs and secrete factors that inhibit the migration of the EVTs. This restriction of EVT migration is in striking contrast to the villous region where EVTs migrate away from the chorion and invade deeply into the decidua. Together, these findings greatly expand our understanding of CTB differentiation in these distinct regions of the human placenta. This knowledge has broad implications for studies of the development, functions, and diseases of the human placenta.

Impact StatementSingle cell RNA-sequencing of distinct regions of the human placenta identifies a smooth chorion-specific cytotrophoblast population responsible for unique functions of the smooth chorion, including acting as a barrier and restricting invasion.
]]></description>
<dc:creator>Marsh, B. P.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Kapzidic, M.</dc:creator>
<dc:creator>Fisher, S.</dc:creator>
<dc:creator>Blelloch, R.</dc:creator>
<dc:date>2022-03-22</dc:date>
<dc:identifier>doi:10.1101/2022.03.21.485195</dc:identifier>
<dc:title><![CDATA[Regionally distinct trophoblast regulate barrier function and invasion in the human placenta]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.25.484773v1?rss=1">
<title>
<![CDATA[
mTORC2 coordinates the leading and trailing edge cytoskeletal programs during neutrophil migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.25.484773v1?rss=1</link>
<description><![CDATA[
By acting both upstream and downstream of biochemical organizers of the cytoskeleton, physical forces function as central integrators of cell shape and movement. Here we use a combination of genetic, pharmacological, and optogenetic perturbations to probe the role of the conserved mechanoresponsive mTORC2 program in neutrophil polarity and motility. We find that the tension-based inhibition of leading edge signals (Rac, F-actin) that underlies protrusion competition is gated by the kinase-independent role of the complex, whereas the mTORC2 kinase arm is essential for regulation of Rho activity and Myosin II-based contraction at the trailing edge. Cells required mTORC2 for spatial and temporal coordination between the front and back polarity programs and persistent migration under confinement. mTORC2 is in a mechanosensory cascade, but membrane stretch did not suffice to stimulate mTORC2 unless the co-input PIP3 was also present. Our work suggests that different signalling arms of mTORC2 regulate spatially and molecularly divergent cytoskeletal programs allowing efficient coordination of neutrophil shape and movement.
]]></description>
<dc:creator>Saha, S.</dc:creator>
<dc:creator>Town, J.</dc:creator>
<dc:creator>Weiner, O.</dc:creator>
<dc:date>2022-03-27</dc:date>
<dc:identifier>doi:10.1101/2022.03.25.484773</dc:identifier>
<dc:title><![CDATA[mTORC2 coordinates the leading and trailing edge cytoskeletal programs during neutrophil migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.28.485852v1?rss=1">
<title>
<![CDATA[
Transient Water Wires Mediate Selective Proton Conduction in Designed Channel Proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.28.485852v1?rss=1</link>
<description><![CDATA[
Summary paragraphSelective proton transport through proteins is essential for forming and utilizing proton gradients in cells. Protons are conducted along hydrogen-bonded "wires" of water molecules and polar sidechains, which, somewhat surprisingly, are often interrupted by dry apolar stretches in the conduction pathways inferred from static protein structures. We hypothesize that protons are conducted through such dry spots by forming transient water wires, often highly correlated with the presence of the excess proton itself in the water wire. To test this hypothesis, we used molecular dynamics simulations to design transmembrane channels with stable water pockets interspersed by apolar segments capable of forming flickering water wires. The minimalist designed channels conduct protons at rates similar to viral proton channels, and they are at least 106-fold more selective for H+ over Na+. These studies inform mechanisms of biological proton conduction and principles for engineering proton-conductive materials.
]]></description>
<dc:creator>Kratochvil, H. T.</dc:creator>
<dc:creator>Watkins, L. C.</dc:creator>
<dc:creator>Mravic, M.</dc:creator>
<dc:creator>Thomaston, J. L.</dc:creator>
<dc:creator>Nicoludis, J. M.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Voth, G. A.</dc:creator>
<dc:creator>DeGrado, W.</dc:creator>
<dc:date>2022-03-28</dc:date>
<dc:identifier>doi:10.1101/2022.03.28.485852</dc:identifier>
<dc:title><![CDATA[Transient Water Wires Mediate Selective Proton Conduction in Designed Channel Proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.31.486626v1?rss=1">
<title>
<![CDATA[
Long-range inhibition synchronizes and updates prefrontal task activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.31.486626v1?rss=1</link>
<description><![CDATA[
Changes in patterns of activity within the medial prefrontal cortex enable rodents, non-human primates, and humans to update their behavior to adapt to changes in the environment, e.g., during cognitive tasks1-5. Within medial prefrontal cortex, inhibitory neurons expressing parvalbumin are important for updating strategies in a rule shift task6-8. Nevertheless, causal mechanisms through which parvalbumin neurons recruit specific circuits to produce prefrontal network dynamics that switch from maintaining to updating task-related patterns of activity remain unknown. Here we identify a new long-range inhibitory projection from prefrontal parvalbumin neurons to the contralateral cortex that mediates this process. Whereas nonspecifically inhibiting all callosal projections does not prevent mice from learning rule shifts, selectively inhibiting these callosal parvalbumin projections disrupts rule shift learning, desynchronizes gamma-frequency activity that is necessary for learning8, and suppresses changes in prefrontal activity patterns that normally accompany rule shifts. Thus, callosal parvalbumin projections switch prefrontal circuits from maintaining to updating behavioral strategies by synchronizing callosal communication and preventing it from inappropriately maintaining outdated neural representations. These findings may explain how deficits in prefrontal parvalbumin neurons and gamma synchrony cause impaired behavioral flexibility in schizophrenia9-10, and identify long-range projections from prefrontal parvalbumin neurons as a key circuit locus for understanding and treating these deficits.
]]></description>
<dc:creator>Cho, K.</dc:creator>
<dc:creator>Shi, J.</dc:creator>
<dc:creator>Sohal, V. S.</dc:creator>
<dc:date>2022-04-01</dc:date>
<dc:identifier>doi:10.1101/2022.03.31.486626</dc:identifier>
<dc:title><![CDATA[Long-range inhibition synchronizes and updates prefrontal task activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.02.486649v1?rss=1">
<title>
<![CDATA[
Active emulsions in living cell membranes driven by contractile stresses and transbilayer coupling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.02.486649v1?rss=1</link>
<description><![CDATA[
The spatiotemporal organisation of proteins and lipids on the cell surface has direct functional consequences for signaling, sorting and endocytosis. Earlier studies have shown that multiple types of membrane proteins including transmembrane proteins that have cytoplasmic actin binding capacity and lipid-tethered GPI-anchored proteins (GPI-APs) form nanoscale clusters driven by active contractile flows generated by the actin cortex. To gain insight into the role of lipids in organizing membrane domains in living cells, we study the molecular interactions that promote the actively generated nanoclusters of GPI-APs and transmembrane proteins. This motivates a theoretical description, wherein a combination of active contractile stresses and transbilayer coupling drive the creation of active emulsions, mesoscale liquid ordered (lo) domains of the GPI-APs and lipids, at temperatures greater than equilibrium lipid-phase segregation. To test these ideas we use spatial imaging of homo-FRET combined with local membrane order and demonstrate that mesoscopic domains enriched in nanoclusters of GPI-APs are maintained by cortical actin activity and transbilayer interactions, and exhibit significant lipid order, consistent with predictions of the active composite model.
]]></description>
<dc:creator>Saha, S.</dc:creator>
<dc:creator>Das, A.</dc:creator>
<dc:creator>Patra, C.</dc:creator>
<dc:creator>Anilkumar, A. A.</dc:creator>
<dc:creator>Sil, P.</dc:creator>
<dc:creator>Mayor, S.</dc:creator>
<dc:creator>Rao, M.</dc:creator>
<dc:date>2022-04-04</dc:date>
<dc:identifier>doi:10.1101/2022.04.02.486649</dc:identifier>
<dc:title><![CDATA[Active emulsions in living cell membranes driven by contractile stresses and transbilayer coupling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.04.487052v1?rss=1">
<title>
<![CDATA[
Microenvironmental engineering improves the self-organization of patterned microvascular networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.04.487052v1?rss=1</link>
<description><![CDATA[
The construction of three-dimensional (3D) microvascular networks with defined structures remains challenging. Emerging bioprinting strategies provide a means of patterning endothelial cells (ECs) into the geometry of 3D microvascular networks, but the microenvironmental cues necessary to promote their self-organization into cohesive and perfusable microvessels are unknown. To this end, we reconstituted microvessel formation in vitro by patterning thin lines of closely packed ECs fully embedded within a 3D extracellular matrix (ECM) and observed how different microenvironmental parameters influenced EC behaviors and their self-organization into microvessels. We found that the inclusion of fibrillar matrices, such as collagen I, into the ECM positively influenced cell condensation into extended geometries such as cords. We also identified the presence of a high molecular weight protein(s) in fetal bovine serum (FBS) that negatively influenced EC condensation. This component destabilized cord structure by promoting cell protrusions and destabilizing cell-cell adhesions. Endothelial cords cultured in the presence of fibrillar collagen and the absence of this protein activity were able to polarize, lumenize, incorporate mural cells, and support fluid flow. These optimized conditions allowed for the construction of branched and perfusable microvascular networks directly from patterned cells in as little as three days. These findings reveal important design principles for future microvascular engineering efforts based on bioprinting techniques.
]]></description>
<dc:creator>Cabral, K. A.</dc:creator>
<dc:creator>Srivastava, V.</dc:creator>
<dc:creator>Coyle, M. C.</dc:creator>
<dc:creator>Stashko, C.</dc:creator>
<dc:creator>Weaver, V.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:date>2022-04-05</dc:date>
<dc:identifier>doi:10.1101/2022.04.04.487052</dc:identifier>
<dc:title><![CDATA[Microenvironmental engineering improves the self-organization of patterned microvascular networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.05.487200v1?rss=1">
<title>
<![CDATA[
Nucleoporins facilitate ORC loading onto chromatin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.05.487200v1?rss=1</link>
<description><![CDATA[
The origin recognition complex (ORC) binds throughout the genome to initiate DNA replication. In metazoans, it is still unclear how ORC is targeted to specific loci to facilitate helicase loading and replication initiation. Here, we performed immunoprecipitations coupled with mass spectrometry for ORC2 in Drosophila embryos. Surprisingly, we found that ORC2 associates with multiple subunits of the Nup107-160 subcomplex of the nuclear pore. Bioinformatic analysis revealed that, relative to all modENCODE factors, nucleoporins are among the most enriched factors at ORC2 binding sites. Critically, depletion of the nucleoporin Elys, a member of the Nup107-160 complex, results in decreased ORC2 loading onto chromatin. Depleting Elys also sensitized cells to replication fork stalling, which could reflect a defect in establishing dormant replication origins. Our work reveals a new connection between ORC, replication initiation and nucleoporins, highlighting a previously unrecognized function of nucleoporins in metazoan replication initiation.
]]></description>
<dc:creator>Richards, L.</dc:creator>
<dc:creator>Lord, C.</dc:creator>
<dc:creator>Benton, M. L.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:creator>Nordman, J. T.</dc:creator>
<dc:date>2022-04-05</dc:date>
<dc:identifier>doi:10.1101/2022.04.05.487200</dc:identifier>
<dc:title><![CDATA[Nucleoporins facilitate ORC loading onto chromatin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.06.487387v1?rss=1">
<title>
<![CDATA[
The {varphi}PA3 Phage Nucleus is Enclosed by a Self-Assembling, 2D Crystalline Lattice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.06.487387v1?rss=1</link>
<description><![CDATA[
A growing number of jumbo bacteriophages, with genomes exceeding 200 kb, have been found to establish a Phage Nucleus--a micron-scale, proteinaceous structure encompassing the replicating phage DNA. Bacteriophage and host proteins associated with replication and transcription are concentrated inside the Phage Nucleus while nucleotide synthesis, translation, and numerous other host and exogenous proteins are effectively excluded, including CRISPR-Cas and restriction endonuclease host defense systems. Here, we show that fragments of the Phage Nucleus isolated from {phi}PA3 infected Pseudomonas aeruginosa cells form a square lattice and demonstrate that the recombinantly purified primary Phage Nuclear Enclosure (PhuN) protein spontaneously assembles into sheets also constructed from a square lattice which we resolve to 3.8 [A] by cryo-EM. Our structure reveals that the flexible termini and large loops mediate adaptable inter-tetramer contacts that drive shell assembly into a C2-symmetric lattice. While the interfaces between subunits are mostly well packed, two of the interfaces are open, forming clear channels that likely have important functional implications for the transport of proteins, mRNA, and small molecules.
]]></description>
<dc:creator>Nieweglowska, E. S.</dc:creator>
<dc:creator>Brilot, A. F.</dc:creator>
<dc:creator>Mendez, M.</dc:creator>
<dc:creator>Baek, M.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Kokontis, C.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Agard, D. A.</dc:creator>
<dc:date>2022-04-06</dc:date>
<dc:identifier>doi:10.1101/2022.04.06.487387</dc:identifier>
<dc:title><![CDATA[The {varphi}PA3 Phage Nucleus is Enclosed by a Self-Assembling, 2D Crystalline Lattice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.08.487649v1?rss=1">
<title>
<![CDATA[
Modeling and mechanical perturbations reveal how spatially regulated anchorage gives rise to spatially distinct mechanics across the mammalian spindle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.08.487649v1?rss=1</link>
<description><![CDATA[
During cell division, the spindle generates force to move chromosomes. In mammals, microtubule bundles called kinetochore-fibers (k-fibers) attach to and segregate chromosomes. To do so, k-fibers must be robustly anchored to the dynamic spindle. We previously developed microneedle manipulation to mechanically challenge k-fiber anchorage, and observed spatially distinct response features revealing the presence of heterogeneous anchorage (Suresh et al. 2020). How anchorage is precisely spatially regulated, and what forces are necessary and sufficient to recapitulate the k-fibers response to force remain unclear. Here, we develop a coarse-grained k-fiber model and combine with manipulation experiments to infer underlying anchorage using shape analysis. By systematically testing different anchorage schemes, we find that forces solely at k-fiber ends are sufficient to recapitulate unmanipulated k-fiber shapes, but not manipulated ones for which lateral anchorage over a 3 m length scale near chromosomes is also essential. Such anchorage robustly preserves k-fiber orientation near chromosomes while allowing pivoting around poles. Anchorage over a shorter length scale cannot robustly restrict pivoting near chromosomes, while anchorage throughout the spindle obstructs pivoting at poles. Together, this work reveals how spatially regulated anchorage gives rise to spatially distinct mechanics in the mammalian spindle, which we propose are key for function.
]]></description>
<dc:creator>Suresh, P.</dc:creator>
<dc:creator>Galstyan, V.</dc:creator>
<dc:creator>Phillips, R.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2022-04-08</dc:date>
<dc:identifier>doi:10.1101/2022.04.08.487649</dc:identifier>
<dc:title><![CDATA[Modeling and mechanical perturbations reveal how spatially regulated anchorage gives rise to spatially distinct mechanics across the mammalian spindle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.08.487704v1?rss=1">
<title>
<![CDATA[
An oligogenic inheritance test detects risk genes and their interactions in congenital heart defects and developmental comorbidities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.08.487704v1?rss=1</link>
<description><![CDATA[
Exome sequencing of thousands of families has revealed many individual risk genes for congenital heart defects (CHD), yet most cases cannot be explained by a single causal mutation. Further, those who carry de novo and inherited mutations in known risk genes often demonstrate variable phenotypes even within the same family, indicating the presence of genetic modifiers. To explore oligogenic causes of CHD without assessing billions of variant combinations, we developed an efficient, simulation-based method to detect gene sets that carry damaging variants in probands at a higher rate than expected given parental genotypes. We implemented this approach in software called Gene Combinations in Oligogenic Disease (GCOD) and applied it to a cohort of 3382 trios with exome sequencing. This analysis detected 353 high-confidence risk genes in 202 pairs that appear together in multiple probands but rarely or never appear in combination in their unaffected parents. Stratifying analyses by specific CHD diagnosis and considering gene combinations of higher orders yielded an additional 244 gene sets. The oligogenic genes we discovered cluster in pathways specific to heart development and suggest new molecular disease mechanisms, such as arylsulfatase activity and de novo nucleotide biosynthesis. Finally, by combining CHD families with an autism spectrum disorder cohort, we were able to detect 925 oligogenic sets transmitted in renal disease, a known co-morbidity of both conditions. As genome sequencing is applied to more families and other disorders, GCOD will enable detection of increasingly large, novel gene combinations, shedding light on combinatorial causes of genetic diseases.
]]></description>
<dc:creator>Pittman, M.</dc:creator>
<dc:creator>Lee, K.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2022-04-10</dc:date>
<dc:identifier>doi:10.1101/2022.04.08.487704</dc:identifier>
<dc:title><![CDATA[An oligogenic inheritance test detects risk genes and their interactions in congenital heart defects and developmental comorbidities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.12.488055v1?rss=1">
<title>
<![CDATA[
Estimating Brain Functional Connectivity from Common Subspace Mapping Between Structural and Functional Connectomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.12.488055v1?rss=1</link>
<description><![CDATA[
Understanding the connection between the brains structural connectivity and its functional connectivity is of immense interest in computational neuroscience. Although some studies have suggested that whole brain functional connectivity is shaped by the underlying structure, the rule by which anatomy constraints brain dynamics remains an open question. In this work, we introduce a computational framework that identifies a joint subspace of eigenmodes for both functional and structural connectomes. We found that a small number of those eigenmodes are sufficient to reconstruct functional connectivity from the structural connectome, thus serving as low-dimensional basis function set. We then develop an algorithm that can estimate the functional eigen spectrum in this joint space from the structural eigen spectrum. By concurrently estimating the joint eigenmodes and the functional eigen spectrum, we can reconstruct a given subjects functional connectivity from their structural connectome. We perform elaborate experiments and demonstrate that the proposed algorithm for estimating functional connectivity from the structural connectome using joint space eigenmodes gives competitive performance as compared to the existing benchmark methods with better interpretability.
]]></description>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2022-04-13</dc:date>
<dc:identifier>doi:10.1101/2022.04.12.488055</dc:identifier>
<dc:title><![CDATA[Estimating Brain Functional Connectivity from Common Subspace Mapping Between Structural and Functional Connectomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.13.488122v1?rss=1">
<title>
<![CDATA[
The cilia enriched oxysterol 7β,27-DHC is required for polycystin activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.13.488122v1?rss=1</link>
<description><![CDATA[
PC-1 and PC-2 form a heteromeric ion channel complex (hereafter called the Polycystin complex) that is abundantly expressed on primary cilia of renal epithelial cells. Mutations within the polycystin complex cause Autosomal Dominant Polycystic Kidney Disease (ADPKD). The Polycystin complex forms a non-selective cation channel, yet the spatial and temporal regulation of the polycystin complex within the ciliary membrane remains poorly understood, partially due to technical limitations posed by the tiny ciliary compartment. Here, we employ our novel assays to functionally reconstitute the polycystin complex in the plasma membrane. Using whole-cell and ciliary patch-clamp recordings we identified a ciliary enriched oxysterol, 7{beta},27-DHC, as a critical component required for activation of the polycystin complex. We identified a novel oxysterol binding pocket in PC-2 using molecular docking simulation. We also identified two amino acids within the PC-2 oxysterol binding pocket, E208 and R581, to be critical for 7{beta},27-DHC dependent polycystin activation in both the plasma membrane and ciliary compartment. Further, we can show that the pharmacological and genetic inhibition of oxysterol synthesis by carbenoxolone (CNX) reduces channel activity in primary cilia. Our findings identified a unique second messenger that regulates the polycystin complex. We hypothesize that cilia-enriched lipids license the polycystin complex to be functional only in the ciliary organelle, thus providing novel insights into the spatial regulation of the polycystin complex. Our results also establish a framework to target the same allosteric regulatory site in the polycystin complex to identify activators of the polycystin channels as novel therapeutic strategies for ADPKD.
]]></description>
<dc:creator>Ha, K.</dc:creator>
<dc:creator>Mundt, N.</dc:creator>
<dc:creator>Bisignano, P.</dc:creator>
<dc:creator>Pinedo, A.</dc:creator>
<dc:creator>Cao, E.</dc:creator>
<dc:creator>Reiter, J.</dc:creator>
<dc:creator>Raleigh, D.</dc:creator>
<dc:creator>Delling, M.</dc:creator>
<dc:date>2022-04-14</dc:date>
<dc:identifier>doi:10.1101/2022.04.13.488122</dc:identifier>
<dc:title><![CDATA[The cilia enriched oxysterol 7β,27-DHC is required for polycystin activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.15.488217v1?rss=1">
<title>
<![CDATA[
Chlamydia trachomatis effector Dre1 interacts with dynactin to reposition host organelles during infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.15.488217v1?rss=1</link>
<description><![CDATA[
Chlamydia trachomatis is an obligate intracellular pathogen that replicates within a specialized membrane-bound compartment, called the inclusion. Chlamydia species express a unique class of effectors, Incs, which are translocated from the bacteria by a Type III secretion system and are inserted into the inclusion membrane where they modulate the host-bacterium interface. C. trachomatis repositions specific host organelles during infection to acquire nutrients and evade host cell surveillance, however the bacterial and host proteins controlling these processes are largely unknown. Here, we identify an interaction between the host dynactin complex and the C. trachomatis Inc CT192 (CTL0444), hereafter named Dre1 for Dynactin Recruiting Effector 1. We show that dynactin is recruited to the inclusion in a Dre1-dependent manner and that loss of Dre1 diminishes the recruitment of specific host organelles, including the centrosome, mitotic spindle, and Golgi apparatus to the inclusion. Inactivation of Dre1 results in decreased C. trachomatis fitness in cell-based assays and in a mouse model of infection. By targeting particular functions of the versatile host dynactin complex, Dre1 facilitates re-arrangement of certain organelles around the growing inclusion. Our work highlights how C. trachomatis employs a single effector to evoke specific, large-scale changes in host cell organization that establish an intracellular replicative niche without globally inhibiting host cellular function.
]]></description>
<dc:creator>Sherry, J.</dc:creator>
<dc:creator>Dolat, L.</dc:creator>
<dc:creator>MacMahon, E.</dc:creator>
<dc:creator>Swaney, D.</dc:creator>
<dc:creator>Bastidas, R.</dc:creator>
<dc:creator>Johnson, J. R.</dc:creator>
<dc:creator>Valdivia, R.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Elwell, C.</dc:creator>
<dc:creator>Engel, J.</dc:creator>
<dc:date>2022-04-18</dc:date>
<dc:identifier>doi:10.1101/2022.04.15.488217</dc:identifier>
<dc:title><![CDATA[Chlamydia trachomatis effector Dre1 interacts with dynactin to reposition host organelles during infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.19.488819v1?rss=1">
<title>
<![CDATA[
Microglial inflammatory activation paradoxically improves glucose tolerance during diet-induced obesity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.19.488819v1?rss=1</link>
<description><![CDATA[
Hypothalamic gliosis associated with high fat diet (HFD) feeding increases susceptibility to hyperphagia and weight gain, and is therefore presumed to promote obesity-associated consequences such as glucose intolerance as well. Nevertheless, the body weight-independent contribution of microglial activation to glucose regulation has not been determined. Here we show that reducing microglial NF-{kappa}B signaling via cell-specific IKK{beta} deletion exacerbates HFD-induced glucose intolerance and insulin resistance despite reducing body weight and adiposity. This effect was associated with reduced activity of hypothalamic glucose sensing neurons. Conversely, a genetic approach to increase microglial inflammatory activity improved glucose tolerance independently of diet in lean rodents. To avoid confounding effects due to chronic alterations to microglial signaling pathways from dietary or genetic interventions, we developed an inducible model of microglial activation using DREADD-based chemogenetics. Gq-coupled GPCR activation rapidly increased microglial calcium levels, cytokine gene expression, and morphological hallmarks of inflammatory activation. In both lean and obese rodents, chemogenetic microglial activation caused a marked improvement in glucose tolerance along with increased activation of hypothalamic glucose sensing neurons, effects abrogated by central blockade of TNF signaling. Thus, while diet-induced microglial activation promotes weight gain, it may also serve an adaptive function--to prevent the deterioration of glucose tolerance associated with obesity, an important consideration for immune-modulating metabolic therapies.
]]></description>
<dc:creator>Douglass, J. D.</dc:creator>
<dc:creator>Valdearcos, M.</dc:creator>
<dc:creator>Ness, K. M.</dc:creator>
<dc:creator>Wyse-Jackson, A.</dc:creator>
<dc:creator>Dorfman, M. D.</dc:creator>
<dc:creator>Frey, J. M.</dc:creator>
<dc:creator>Niraula, A.</dc:creator>
<dc:creator>Banerjee, J.</dc:creator>
<dc:creator>Fasnacht, R.</dc:creator>
<dc:creator>Robblee, M.</dc:creator>
<dc:creator>Koliwad, S. K.</dc:creator>
<dc:creator>Thaler, J.</dc:creator>
<dc:date>2022-04-20</dc:date>
<dc:identifier>doi:10.1101/2022.04.19.488819</dc:identifier>
<dc:title><![CDATA[Microglial inflammatory activation paradoxically improves glucose tolerance during diet-induced obesity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.20.488974v1?rss=1">
<title>
<![CDATA[
Genome-wide CRISPR screens of T cell exhaustion identify chromatin remodeling factors that limit T cell persistence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.20.488974v1?rss=1</link>
<description><![CDATA[
T cell exhaustion limits anti-tumor immunity, but the molecular determinants of this process remain poorly understood. Using a chronic antigen stimulation assay, we performed genome-wide CRISPR/Cas9 screens to systematically discover genetic regulators of T cell exhaustion, which identified an enrichment of epigenetic factors. In vivo CRISPR screens in murine and human tumor models demonstrated that perturbation of several epigenetic regulators, including members of the INO80 and BAF chromatin remodeling complexes, improved T cell persistence in tumors. In vivo paired CRISPR perturbation and single-cell RNA sequencing revealed distinct transcriptional roles of each complex and that depletion of canonical BAF complex members, including Arid1a, resulted in the maintenance of an effector program and downregulation of terminal exhaustion-related genes in tumor-infiltrating T cells. Finally, Arid1a-depletion limited the global acquisition of chromatin accessibility associated with T cell exhaustion and led to improved anti-tumor immunity after adoptive cell therapy. In summary, we provide a comprehensive atlas of the genetic regulators of T cell exhaustion and demonstrate that modulation of the epigenetic state of T cell exhaustion can improve T cell responses in cancer immunotherapy.
]]></description>
<dc:creator>Belk, J.</dc:creator>
<dc:creator>Yao, W.</dc:creator>
<dc:creator>Ly, N.</dc:creator>
<dc:creator>Freitas, K.</dc:creator>
<dc:creator>Chen, Y.-T.</dc:creator>
<dc:creator>Shi, Q.</dc:creator>
<dc:creator>Valencia, A.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Kale, N.</dc:creator>
<dc:creator>Yost, K.</dc:creator>
<dc:creator>Duffy, C.</dc:creator>
<dc:creator>Hwee, M.</dc:creator>
<dc:creator>Miao, Z.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Mackall, C.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Carnevale, J.</dc:creator>
<dc:creator>Vardhana, S.</dc:creator>
<dc:creator>Satpathy, A.</dc:creator>
<dc:date>2022-04-21</dc:date>
<dc:identifier>doi:10.1101/2022.04.20.488974</dc:identifier>
<dc:title><![CDATA[Genome-wide CRISPR screens of T cell exhaustion identify chromatin remodeling factors that limit T cell persistence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.28.489942v1?rss=1">
<title>
<![CDATA[
Immediate myeloid depot for SARS-CoV-2 in the human lung 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.28.489942v1?rss=1</link>
<description><![CDATA[
In the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, considerable focus has been placed on a model of viral entry into host epithelial populations, with a separate focus upon the responding immune system dysfunction that exacerbates or causes disease. We developed a precision-cut lung slice model to investigate very early host-viral pathogenesis and found that SARS-CoV-2 had a rapid and specific tropism for myeloid populations in the human lung. Infection of alveolar macrophages was partially dependent upon their expression of ACE2, and the infections were productive for amplifying virus, both findings which were in contrast with their neutralization of another pandemic virus, Influenza A virus (IAV). Compared to IAV, SARS-CoV-2 was extremely poor at inducing interferon-stimulated genes in infected myeloid cells, providing a window of opportunity for modest titers to amplify within these cells. Endotracheal aspirate samples from humans with the acute respiratory distress syndrome (ARDS) from COVID-19 confirmed the lung slice findings, revealing a persistent myeloid depot. In the early phase of SARS-CoV-2 infection, myeloid cells may provide a safe harbor for the virus with minimal immune stimulatory cues being generated, resulting in effective viral colonization and quenching of the immune system.
]]></description>
<dc:creator>Magnen, M.</dc:creator>
<dc:creator>You, R.</dc:creator>
<dc:creator>Rao, A. A.</dc:creator>
<dc:creator>Davis, R. T.</dc:creator>
<dc:creator>Rodriguez, L.</dc:creator>
<dc:creator>Simoneau, C. R.</dc:creator>
<dc:creator>Hysenaj, L.</dc:creator>
<dc:creator>Hu, K. H.</dc:creator>
<dc:creator>The UCSF COMET Consortium,</dc:creator>
<dc:creator>Love, C.</dc:creator>
<dc:creator>Woodruff, P. G.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:creator>Hendrickson, C. M.</dc:creator>
<dc:creator>Calfee, C. S.</dc:creator>
<dc:creator>Matthay, M. A.</dc:creator>
<dc:creator>Roose, J. P.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Langelier, C. R.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Looney, M. R.</dc:creator>
<dc:date>2022-04-29</dc:date>
<dc:identifier>doi:10.1101/2022.04.28.489942</dc:identifier>
<dc:title><![CDATA[Immediate myeloid depot for SARS-CoV-2 in the human lung]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.01.489793v1?rss=1">
<title>
<![CDATA[
A transcriptional mechanism involving R-loop, m6A modification and RNA abasic sites regulates an enhancer RNA of APOE. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.01.489793v1?rss=1</link>
<description><![CDATA[
The DNA genetic code and the RNA regulatory code determine phenotypes from gene expression to disease susceptibility. DNA sequence variants lead to phenotypic differences among individuals, while within an individual, RNA dynamically confers cell identity and responds to cellular and environmental signals. To provide regulation for different cell types and conditions, the nucleotides of RNA are modified by hundreds of chemical reactions, and RNA folds into innumerable shapes. To decipher the RNA regulatory code is to understand how RNA sequence and structure respond to cellular needs. Here, we deciphered one part of the RNA code where RNA abasic sites in R-loops regulate transcription by pausing RNA polymerase II. We uncovered an enhancer RNA, AANCR, that regulates the transcription and expression of APOE. When AANCR is folded into an R-loop, which is then modified by N6-adenine methylation and N-glycosidic cleavage, it is a partially transcribed nonfunctional enhancer and APOE is not expressed. In contrast, in some cell types and under stress, AANCR does not form a stable R-loop as its sequence is not modified, so it is transcribed into a full-length enhancer that promotes APOE expression. By genetic analysis, we confirmed that AANCR regulates APOE expression. DNA sequence variants in AANCR are associated with APOE expression and also with Alzheimers disease. Our data show that DNA and RNA sequence and structure jointly regulate gene expression that influence disease risk.

HighlightsO_LIm6A, RNA abasic sites and R-loops jointly regulate transcription by pausing RNA Polymerase II.
C_LIO_LIAn enhancer RNA regulates APOE expression.
C_LIO_LIEnhancer RNA of APOE modifies susceptibility to Alzheimers disease.
C_LI
]]></description>
<dc:creator>Watts, J. A.</dc:creator>
<dc:creator>Grunseich, C.</dc:creator>
<dc:creator>Rodriguez, Y.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Burdick, J. T.</dc:creator>
<dc:creator>Bruzel, A.</dc:creator>
<dc:creator>Crouch, R. J.</dc:creator>
<dc:creator>Mahley, R. W.</dc:creator>
<dc:creator>Wilson, S.</dc:creator>
<dc:creator>Cheung, V. G.</dc:creator>
<dc:date>2022-05-01</dc:date>
<dc:identifier>doi:10.1101/2022.05.01.489793</dc:identifier>
<dc:title><![CDATA[A transcriptional mechanism involving R-loop, m6A modification and RNA abasic sites regulates an enhancer RNA of APOE.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.04.490528v1?rss=1">
<title>
<![CDATA[
Ribotoxic collisions on CAG expansions disrupt proteostasis and stress responses in Huntington's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.04.490528v1?rss=1</link>
<description><![CDATA[
Huntingtons disease (HD) is a devastating neurodegenerative disorder caused by CAG trinucleotide repeat expansions encoding a polyglutamine (polyQ) tract in the Huntingtin (HTT) gene1. Although mutant HTT (mHTT) protein tends to aggregate, the exact causes of neurotoxicity in HD remain unclear2. Here we show that altered elongation kinetics on CAG expansions cause ribosome collisions that trigger ribotoxicity, proteotoxicity and maladaptive stress responses. CAG expansions cause an elongation rate conflict during HTT translation, when ribosomes rapidly decoding the optimal polyQ encounter a flanking slowly-decoded polyproline tract. The ensuing ribosome collisions lead to premature termination and release of aggregation-prone mHTT fragments. Due to the presence of a stress-responsive upstream open reading frame (uORF), HTT translation and aggregation are limited under normal conditions but enhanced under stress, seeding a vicious cycle of dysfunction. mHTT further exacerbates ribotoxicity by progressively sequestering eIF5A, a key regulator of translation elongation, polyamine metabolism and stress responses. eIF5A depletion in HD cells leads to widespread ribosome pausing on eIF5A-dependent sites, impaired cotranslational proteostasis, disrupted polyamine metabolism and maladaptive stress responses. Importantly, drugs that reduce translation initiation attenuate ribosome collisions and mitigate this escalating cascade of ribotoxic stress and dysfunction in HD.
]]></description>
<dc:creator>Aviner, R.</dc:creator>
<dc:creator>Lee, T.-T.</dc:creator>
<dc:creator>Masto, V. B.</dc:creator>
<dc:creator>Gestaut, D.</dc:creator>
<dc:creator>Li, K. H.</dc:creator>
<dc:creator>Andino, R. P.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:date>2022-05-05</dc:date>
<dc:identifier>doi:10.1101/2022.05.04.490528</dc:identifier>
<dc:title><![CDATA[Ribotoxic collisions on CAG expansions disrupt proteostasis and stress responses in Huntington's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.07.491000v1?rss=1">
<title>
<![CDATA[
Multi-tasking Deep Network for Tinnitus Classification and Severity Prediction from Multimodal Structural Images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.07.491000v1?rss=1</link>
<description><![CDATA[
Subjective tinnitus is an auditory phantom perceptual disorder without an objective biomarker. Fast and efficient diagnostic tools will advance clinical practice by detecting or confirming the condition, tracking change in severity, and monitoring treatment response. Motivated by evidence of subtle anatomical or functional morphological information in magnetic resonance images (MRI) of the brain, we examined data-driven machine learning methods for joint tinnitus classification (tinnitus or no tinnitus) and tinnitus severity prediction. We propose a deep multi-task multi-modal framework for joint functionalities using structural MRI (sMRI) data. To leverage cross-information multimodal neuroimaging data, we integrated two modalities of 3-dimensional sMRI - T1 weighted (T1w) and T2 weighted (T2w) images. To explore the key components in the MR images that drove task performance, we segmented both T1w and T2w images into three different components - cerebrospinal fluid (CSF), grey matter (GM) and white matter (WM), and examined performance of each segmented image. Results demonstrate that our multimodal framework capitalizes on the information across both modalities (T1w and T2w) for the joint task of tinnitus classification and severity prediction. Our model outperforms existing learning-based and conventional methods in terms of accuracy, sensitivity, specificity, and negative predictive value.
]]></description>
<dc:creator>Lin, C.-T.</dc:creator>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Hinkley, L. B.</dc:creator>
<dc:creator>Dale, C. L.</dc:creator>
<dc:creator>Souza, A. C.</dc:creator>
<dc:creator>Sabes, J. H.</dc:creator>
<dc:creator>Hess, C. P.</dc:creator>
<dc:creator>Adams, M. E.</dc:creator>
<dc:creator>Cheung, S. W.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2022-05-08</dc:date>
<dc:identifier>doi:10.1101/2022.05.07.491000</dc:identifier>
<dc:title><![CDATA[Multi-tasking Deep Network for Tinnitus Classification and Severity Prediction from Multimodal Structural Images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.10.491392v1?rss=1">
<title>
<![CDATA[
Accelerating the clock: Interconnected speedup of energetic and molecular dynamics during aging in cultured human cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.10.491392v1?rss=1</link>
<description><![CDATA[
To understand how organisms age, we need reliable multimodal molecular data collected at high temporal resolution, in specific cell types, across the lifespan. We also need interpretative theory that connects aging with basic mechanisms and physiological tradeoffs. Here we leverage a simple cellular replicative aging system combined with mathematical theory to address organismal aging. We used cultured primary human fibroblasts from multiple donors to molecularly and energetically profile entire effective lifespans of up to nine months. We generated high-density trajectories of division rates, telomere shortening, DNA methylation, RNAseq, secreted proteins/cytokines and cell-free DNA, in parallel with bioenergetic trajectories of ATP synthesis rates derived from both mitochondrial oxidative phosphorylation and glycolysis, reflecting total cellular mass-specific metabolic rate (MR). By comparing our cell culture data to data from cells in the body we uncover three fundamental speedups, or rescalings, of MR and molecular aging markers. To explain these rescalings we deploy the allometric theory of metabolism which predicts that the rate of biological aging is related to an organisms size, MR, and the partitioning of energetic resources between growth and maintenance processes. Extending this theory we report three main findings: 1) human cells isolated from the body with faster rates of growth allocate a substantially smaller fraction of their energy budget to maintenance, and correspondingly age 50-300x faster based on multiple molecular markers. 2) Over the course of the cellular lifespan, primary human fibroblasts acquire a >100-fold hypermetabolic phenotype characterized by increased maintenance costs, and associated with increased mtDNA genome density, upregulation of senescence-associated extracellular secretion, and induction of maintenance-related transcriptional programs. 3) Finally, manipulating MR with mitochondria-targeted metabolic, genetic, and pharmacological perturbations predictably altered the molecular rate of aging, providing experimental evidence for the interplay of MR and aging in a human system. These data highlight the key role that the partitioning of energetic resources between growth and maintenance/repair processes plays in cellular aging, and converge with predictions of cross-species metabolic theory indicating that energy metabolism governs how human cells age.

Significance StatementHow cells age is of fundamental importance to understanding the diversity of mammalian lifespans and the wide variation in human aging trajectories. By aging primary human fibroblasts over several months in parallel with multi-omics and energetic profiling, we find that as human cells age and progressively divide more slowly, surprisingly, they progressively consume energy faster. By manipulating cellular metabolic rates, we confirm that the higher the cellular metabolic rate, the faster cells experience telomere shortening and epigenetic aging - a speedup phenotype consistent with allometric scaling theory. By modeling robust energetic and molecular aging trajectories across donors and experimental conditions, we find that independent of cell division rates, molecular aging trajectories are predicted by the partitioning of the energy budget between growth and maintenance processes. These results integrate molecular and energetic drivers of aging and therefore have important long-term implications to understand biological aging phenomena ranging from cellular senescence to human longevity.
]]></description>
<dc:creator>Sturm, G.</dc:creator>
<dc:creator>Bobba-Alves, N.</dc:creator>
<dc:creator>Tumasian, R. A.</dc:creator>
<dc:creator>Michelson, J.</dc:creator>
<dc:creator>Ferrucci, L.</dc:creator>
<dc:creator>Kempes, C. P.</dc:creator>
<dc:creator>Picard, M.</dc:creator>
<dc:date>2022-05-12</dc:date>
<dc:identifier>doi:10.1101/2022.05.10.491392</dc:identifier>
<dc:title><![CDATA[Accelerating the clock: Interconnected speedup of energetic and molecular dynamics during aging in cultured human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.13.491826v1?rss=1">
<title>
<![CDATA[
Perturbation of ATP6V0A1 as a Driver and Druggable Target for Renal Proximal Tubular Cellular Injury in Nephropathic Cystinosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.13.491826v1?rss=1</link>
<description><![CDATA[
Understanding the unique susceptibility of the human kidney to pH dysfunction and injury in cystinosis is paramount to developing new therapies to preserve renal function. Renal proximal tubular epithelial cells (RPTECs) and fibroblasts isolated from patients with cystinosis were transcriptionally profiled. Lysosomal fractionation, immunoblotting, confocal microscopy, intracellular pH, TEM, mitochondrial stress test, and membrane integrity assays were performed for validation. CRISPR, CTNS -/- RPTECs were generated. Alterations in cell stress, pH, autophagic turnover, and mitochondrial energetics highlighted key changes in the vacuolar (V)-ATPases in patient-derived and CTNS-/- RPTECs. ATP6V0A1 was significantly downregulated in cystinosis and highly co-regulated with loss of CTNS. Correction of ATP6V0A1 rescued cell stress and mitochondrial function. Treatment of CTNS -/- RPTECs with antioxidants astaxanthin (ATX) induced ATP6V0A1 expression and improved autophagosome turnover and mitochondrial integrity.

In conclusion, our exploratory transcriptional and in vitro cellular and functional studies confirm that loss of cystinosin in RPTECs, results in a reduction in ATP6V0A1 expression, with changes in intracellular pH, mitochondrial integrity, mitochondrial function, and autophagosome-lysosome clearance. The novel findings are ATP6V0A1s role in cystinosis-associated renal pathology and among other antioxidants, ATX specifically upregulated ATP6V0A1, improved autophagosome turnover or reduced autophagy and mitochondrial integrity. This is a pilot study highlighting a novel mechanism of tubular injury in cystinosis and requires further study in animal models to clarify its utility in clinical settings.
]]></description>
<dc:creator>Sur, S.</dc:creator>
<dc:creator>Kerwin, M.</dc:creator>
<dc:creator>Pineda, S.</dc:creator>
<dc:creator>Sansanwal, P.</dc:creator>
<dc:creator>Sigdel, T. K.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Sarwal, M. M.</dc:creator>
<dc:date>2022-05-13</dc:date>
<dc:identifier>doi:10.1101/2022.05.13.491826</dc:identifier>
<dc:title><![CDATA[Perturbation of ATP6V0A1 as a Driver and Druggable Target for Renal Proximal Tubular Cellular Injury in Nephropathic Cystinosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.16.492147v1?rss=1">
<title>
<![CDATA[
MFGE8 links absorption of dietary fatty acids with catabolism of enterocyte lipid stores through HNF4 γ- dependent transcription of CES enzymes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.16.492147v1?rss=1</link>
<description><![CDATA[
Enterocytes modulate the extent of postprandial lipemia, a potent risk factor for developing atherosclerotic disease, by storing dietary fats in cytoplasmic lipid droplets (cLDs). We have previously demonstrated that the integrin ligand MFGE8 links absorption of dietary fats with activation of triglyceride (TG) hydrolases that catabolize cLDs for chylomicron production. The hydrolase(s) responsible for mobilization of TG from diet-derived cLDs is unknown though recent evidence indicates that this process is independent of the canonical pathway of TG hydrolysis mediated by ATGL. Here we identify CES1D as the key hydrolase downstream of the MFGE8-v{beta}5 integrin pathway that regulates catabolism of diet-drive cLDs. Mfge8 KO enterocytes have reduced CES1D transcript and protein levels and reduced protein levels of the transcription factor HNF4{gamma}. Mice KO for Ces1d or Hnf4{gamma} have decreased enterocyte TG hydrolase activity coupled with retention of TG in cLDs. Mechanistically, MFGE8-dependent fatty acid uptake through CD36 leads to stabilization of HNF4{gamma} protein levels; HNF4{gamma} then increases Ces1d transcription. Our work identifies a regulatory network by which MFGE8 and v{beta}5 regulate the severity of postprandial lipemia by linking dietary fat absorption with protein stabilization of a transcription factor that increases expression of enterocyte TG hydrolases that catabolize diet-derived cLDs.
]]></description>
<dc:creator>Datta, R.</dc:creator>
<dc:creator>Gholampour, M. A.</dc:creator>
<dc:creator>Yang, C. D.</dc:creator>
<dc:creator>Volk, R.</dc:creator>
<dc:creator>Lin, S.</dc:creator>
<dc:creator>Podolsky, M. J.</dc:creator>
<dc:creator>Arnold, T.</dc:creator>
<dc:creator>Rieder, F.</dc:creator>
<dc:creator>Zaro, B. W.</dc:creator>
<dc:creator>Verzi, M.</dc:creator>
<dc:creator>Lehner, R.</dc:creator>
<dc:creator>Abumrad, N.</dc:creator>
<dc:creator>Lizama, C. O.</dc:creator>
<dc:creator>Atabai, K.</dc:creator>
<dc:date>2022-05-18</dc:date>
<dc:identifier>doi:10.1101/2022.05.16.492147</dc:identifier>
<dc:title><![CDATA[MFGE8 links absorption of dietary fatty acids with catabolism of enterocyte lipid stores through HNF4 γ- dependent transcription of CES enzymes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.19.491203v1?rss=1">
<title>
<![CDATA[
Long-term functional regeneration of radiation damaged salivary glands through delivery of a neurogenic hydrogel 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.19.491203v1?rss=1</link>
<description><![CDATA[
Salivary gland acinar cells are severely depleted after radiotherapy for head and neck cancer, leading to loss of saliva and extensive oro-digestive complications. With no regenerative therapies available, organ dysfunction is irreversible. Here using the adult murine system, we demonstrate radiation-damaged salivary glands can be functionally regenerated via sustained delivery of the neurogenic muscarinic receptor agonist, cevimeline. We show that endogenous gland repair coincides with increased nerve activity and acinar cell division that is limited to the first week post-radiation, with extensive acinar cell degeneration, dysfunction and cholinergic denervation occurring thereafter. However, we discovered that mimicking cholinergic muscarinic input via sustained local delivery of a cevimeline-alginate hydrogel was sufficient to regenerate innervated acini and retain physiological saliva secretion at non-irradiated levels over the long-term (> 3 months). Thus, we reveal a novel regenerative approach for restoring epithelial organ structure and function that has significant implications for human patients.

TeaserNovel application of an injectable neurogenic-based hydrogel for restoring the structure and function of radiation-damaged tissue.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Sudiwala, S.</dc:creator>
<dc:creator>Berthoin, L.</dc:creator>
<dc:creator>May, A. J.</dc:creator>
<dc:creator>Mohabbat, S.</dc:creator>
<dc:creator>Gaylord, E. A.</dc:creator>
<dc:creator>Cruz Pacheco, N.</dc:creator>
<dc:creator>Lombaert, I. A. M.</dc:creator>
<dc:creator>Jeon, O.</dc:creator>
<dc:creator>Alsberg, E.</dc:creator>
<dc:creator>Bahney, C. S.</dc:creator>
<dc:creator>Knox, S. M.</dc:creator>
<dc:date>2022-05-19</dc:date>
<dc:identifier>doi:10.1101/2022.05.19.491203</dc:identifier>
<dc:title><![CDATA[Long-term functional regeneration of radiation damaged salivary glands through delivery of a neurogenic hydrogel]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.21.492887v1?rss=1">
<title>
<![CDATA[
Two therapeutic CRISPR/Cas9 gene editing approaches revert FTD/ALS cellular pathology caused by a C9orf72 repeat expansion mutation in patient derived cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.21.492887v1?rss=1</link>
<description><![CDATA[
CRISPR gene editing holds promise to cure or arrest genetic disease, if we can find and implement curative edits reliably, safely and effectively. Expansion of a hexanucleotide repeat in C9orf72 is the leading known genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). We evaluated three approaches to editing the mutant C9orf72 gene for their ability to correct pathology in neurons derived from patient iPSCs: excision of the repeat region, excision of the mutant allele, and excision of regulatory region exon 1A. All three approaches normalized RNA abnormalities and TDP-43 pathology, but only repeat excision and mutant allele excision completely eliminated pathologic dipeptide repeats. Our work sheds light on the complex regulation of the C9orf72 gene and suggests that because of sense and anti-sense transcription, silencing a single regulatory region may not reverse all pathology. Our work also provides a roadmap for evaluating CRISPR gene correction using patient iPSCs.
]]></description>
<dc:creator>Sckaff, M.</dc:creator>
<dc:creator>Gill, K.</dc:creator>
<dc:creator>Sachdev, A.</dc:creator>
<dc:creator>Birk, A. M.</dc:creator>
<dc:creator>Aladesuyi Arogundade, O.</dc:creator>
<dc:creator>Watry, H. L.</dc:creator>
<dc:creator>Keough, K. C.</dc:creator>
<dc:creator>Tsai, Y.-C.</dc:creator>
<dc:creator>Ziegle, J.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:creator>Clelland, C. D.</dc:creator>
<dc:date>2022-05-21</dc:date>
<dc:identifier>doi:10.1101/2022.05.21.492887</dc:identifier>
<dc:title><![CDATA[Two therapeutic CRISPR/Cas9 gene editing approaches revert FTD/ALS cellular pathology caused by a C9orf72 repeat expansion mutation in patient derived cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.23.493138v1?rss=1">
<title>
<![CDATA[
Reconstitution of the SARS-CoV-2 ribonucleosome provides insights into genomic RNA packaging and regulation by phosphorylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.23.493138v1?rss=1</link>
<description><![CDATA[
The nucleocapsid (N) protein of coronaviruses is responsible for compaction of the [~]30-kb RNA genome in the [~]100-nm virion. Cryo-electron tomography suggests that each virion contains 35-40 viral ribonucleoprotein (vRNP) complexes, or ribonucleosomes, arrayed along the genome. There is, however, little mechanistic understanding of the vRNP complex. Here, we show that N protein, when combined with viral RNA fragments in vitro, forms cylindrical 15-nm particles similar to the vRNP structures observed within coronavirus virions. These vRNPs form in the presence of stem-loop-containing RNA and depend on regions of N protein that promote protein-RNA and protein-protein interactions. Phosphorylation of N protein in its disordered serine/arginine (SR) region weakens these interactions and disrupts vRNP assembly. We propose that unmodified N binds stem-loop-rich regions in genomic RNA to form compact vRNP complexes within the nucleocapsid, while phosphorylated N maintains uncompacted viral RNA to promote the proteins transcriptional function.
]]></description>
<dc:creator>Carlson, C.</dc:creator>
<dc:creator>Adly, A.</dc:creator>
<dc:creator>Bi, M.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Morgan, D. O.</dc:creator>
<dc:date>2022-05-24</dc:date>
<dc:identifier>doi:10.1101/2022.05.23.493138</dc:identifier>
<dc:title><![CDATA[Reconstitution of the SARS-CoV-2 ribonucleosome provides insights into genomic RNA packaging and regulation by phosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.24.493171v1?rss=1">
<title>
<![CDATA[
The transcription regulator Lmo3 is required for cell fate specification in the external globus pallidus. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.24.493171v1?rss=1</link>
<description><![CDATA[
The external globus pallidus (GPe) is an essential component of the basal ganglia, a group of subcortical nuclei that are involved in control of action. Changes in the firing of GPe neurons are associated with both passive and active body movements. Aberrant activity of GPe neurons has been linked to motor symptoms of a variety of movement disorders, such as Parkinsons Disease, Huntingtons disease and dystonia. Recent studies have helped delineate functionally distinct sub types of GABAergic GPe projection neurons. However, little remains known about specific molecular mechanisms underlying the development of GPe neuronal subtypes. We show that the transcriptional regulator Lmo3 is required for the development of medial ganglionic eminence derived Nkx2.1+ and PV+ GPe neurons, but not FoxP2+ neurons or Npas1+ neurons. As a consequence of the reduction in PV+ neurons, Lmo3-null mice have a reduced pallidal input to the subthalamic nucleus.

SIGNIFICANCE STATEMENTThe external globus pallidus (GPe) is a critical component of the basal ganglia and can coordinate neuronal activity across the basal ganglia by virtue of its widespread projections to almost all other basal ganglia nuclei. Aberrant activity of GPe neurons has been linked to motor symptoms of a wide variety of movement disorders. Recent advances have delineated functionally distinct sub types of GABAergic GPe projection neurons. However, little remains known about molecular mechanisms underlying their development. Here, we demonstrate that the transcription regulator Lmo3 is required for the development of specific subtypes of GPe neurons, and for their appropriate connectivity with other parts of the basal ganglia.
]]></description>
<dc:creator>Biswas, S.</dc:creator>
<dc:creator>Chan, C. S.</dc:creator>
<dc:creator>Rubenstein, J. L. R.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:date>2022-05-24</dc:date>
<dc:identifier>doi:10.1101/2022.05.24.493171</dc:identifier>
<dc:title><![CDATA[The transcription regulator Lmo3 is required for cell fate specification in the external globus pallidus.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.24.493310v1?rss=1">
<title>
<![CDATA[
A preprocessing and functional connectivity analysis pipeline for mesoscale calcium imaging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.24.493310v1?rss=1</link>
<description><![CDATA[
To improve  bench-to-bedside translation, it is integral that knowledge flow bidirectionally--from animal models to humans, and vice versa. This requires common analytical frameworks, as well as open software and data sharing practices. We share a new pipeline (and test dataset) for the preprocessing of wide-field optical fluorescence imaging data--an emerging mode applicable in animal models--as well as results from a functional connectivity and graph theory analysis inspired by recent work in the human neuroimaging field. The approach is demonstrated using a dataset comprised of two test-cases: (1) data from animals imaged during awake and anesthetized conditions with excitatory neurons labeled, and (2) data from awake animals with different genetically encoded fluorescent labels that target either excitatory neurons or inhibitory interneuron subtypes. Both seed-based connectivity and graph theory measures (global efficiency, transitivity, modularity, and characteristic path-length) are shown to be useful in quantifying differences between wakefulness states and cell populations. Wakefulness state and cell type show widespread effects on canonical network connectivity with variable frequency band dependence. Differences between excitatory neurons and inhibitory interneurons are observed, with somatostatin expressing inhibitory interneurons emerging as notably dissimilar from parvalbumin and vasoactive polypeptide expressing cells. In sum, we demonstrate that our pipeline can be used to examine brain state and cell-type differences in mesoscale imaging data, aiding translational neuroscience efforts. In line with open science practices, we freely release the pipeline and data to encourage other efforts in the community.
]]></description>
<dc:creator>O'Connor, D.</dc:creator>
<dc:creator>Mandino, F.</dc:creator>
<dc:creator>Shen, X.</dc:creator>
<dc:creator>Horien, C.</dc:creator>
<dc:creator>Ge, X.</dc:creator>
<dc:creator>Herman, P.</dc:creator>
<dc:creator>Crair, M.</dc:creator>
<dc:creator>Papademetris, X.</dc:creator>
<dc:creator>Lake, E. M.</dc:creator>
<dc:creator>Constable, R. T.</dc:creator>
<dc:date>2022-05-25</dc:date>
<dc:identifier>doi:10.1101/2022.05.24.493310</dc:identifier>
<dc:title><![CDATA[A preprocessing and functional connectivity analysis pipeline for mesoscale calcium imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.31.494136v1?rss=1">
<title>
<![CDATA[
Fatty acid synthesis knockdown promotes biofilm wrinkling and inhibits sporulation in Bacillus subtilis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.31.494136v1?rss=1</link>
<description><![CDATA[
Many bacterial species typically live in complex three-dimensional biofilms, yet little is known about systematic changes to gene function between non-biofilm and biofilm lifestyles. Here, we created a CRISPRi library of knockdown strains covering all known essential genes in the biofilm-forming Bacillus subtilis strain 3610. We show that gene essentiality is largely conserved between liquid and surface growth and between two media. We developed an image analysis algorithm to quantify biofilm colony wrinkling, which identified strains with high or low levels of wrinkling that were uncorrelated with extracellular matrix gene expression. We also designed a high-throughput screen for sensitive quantification of sporulation efficiency and performed the first screens of sporulation during essential gene knockdown. We found that all basal knockdowns of essential genes were competent for sporulation in a sporulation-inducing medium, but certain strains exhibited reduced sporulation efficiency in LB, a medium with generally lower levels of sporulation. Knockdown of fatty acid synthesis increased wrinkling and inhibited sporulation. These results highlight the importance of essential genes in biofilm structure and sporulation/germination and suggest a previously unappreciated and multifaceted role for fatty acid synthesis in bacterial lifestyles and developmental processes.

Abstract ImportanceFor many bacteria, life typically involves growth in dense, three-dimensional communities called biofilms that contain cells with differentiated roles and are held together by extracellular matrix. To examine how gene function varies between non-biofilm and biofilm growth, we created a comprehensive library of strains using CRISPRi to knockdown expression of each essential gene in the model species Bacillus subtilis 3610, which can develop into a wrinkled biofilm structure or a spore capable of surviving harsh environments. This library enabled us to determine when gene essentiality depends on growth conditions. We also developed high-throughput assays and computational algorithms to identify essential genes involved in biofilm wrinkling and sporulation. Knockdown of fatty acid synthesis increased the density of wrinkles, and also inhibited sporulation in a medium with generally lower sporulation levels. These findings indicate that essential processes such as fatty acid synthesis can play important and multifaceted roles in bacterial development.
]]></description>
<dc:creator>Arjes, H. A.</dc:creator>
<dc:creator>Gui, H.</dc:creator>
<dc:creator>Porter, R.</dc:creator>
<dc:creator>Atolia, E.</dc:creator>
<dc:creator>Peters, J.</dc:creator>
<dc:creator>Gross, C.</dc:creator>
<dc:creator>Kearns, D. B.</dc:creator>
<dc:creator>Huang, K. A.</dc:creator>
<dc:date>2022-05-31</dc:date>
<dc:identifier>doi:10.1101/2022.05.31.494136</dc:identifier>
<dc:title><![CDATA[Fatty acid synthesis knockdown promotes biofilm wrinkling and inhibits sporulation in Bacillus subtilis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.31.494205v1?rss=1">
<title>
<![CDATA[
Physical and functional convergence of the autism risk genes Scn2a and Ank2 in neocortical pyramidal cell dendrites 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.31.494205v1?rss=1</link>
<description><![CDATA[
Dysfunction in sodium channels and their ankyrin scaffolding partners have both been implicated in neurodevelopmental disorders, including autism spectrum disorder (ASD). In particular, the genes SCN2A, which encodes the sodium channel NaV1.2, and ANK2, which encodes ankyrin-B, have strong ASD association. Recent studies indicate that ASD-associated haploinsufficiency in Scn2a impairs dendritic excitability and synaptic function in neocortical pyramidal cells, but how NaV1.2 is anchored within dendritic regions is unknown. Here, we show that ankyrin-B is essential for scaffolding NaV1.2 to the dendritic membrane of mouse neocortical neurons, and that haploinsufficiency of Ank2 phenocopies intrinsic dendritic excitability and synaptic deficits observed in Scn2a+/- conditions. Thus, these results establish a direct, convergent link between two major ASD risk genes and reinforce an emerging framework suggesting that neocortical pyramidal cell dendritic dysfunction can be etiological to neurodevelopmental disorder pathophysiology.
]]></description>
<dc:creator>Nelson, A. D.</dc:creator>
<dc:creator>Catalfio, A. M.</dc:creator>
<dc:creator>Gupta, J. M.</dc:creator>
<dc:creator>Min, L.</dc:creator>
<dc:creator>Caballero-Floran, R. N.</dc:creator>
<dc:creator>Dean, K. P.</dc:creator>
<dc:creator>Elvira, C. C.</dc:creator>
<dc:creator>Derderian, K. D.</dc:creator>
<dc:creator>Kyoung, H.</dc:creator>
<dc:creator>Sahagun, A.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Bender, K.</dc:creator>
<dc:creator>Jenkins, P. M.</dc:creator>
<dc:date>2022-05-31</dc:date>
<dc:identifier>doi:10.1101/2022.05.31.494205</dc:identifier>
<dc:title><![CDATA[Physical and functional convergence of the autism risk genes Scn2a and Ank2 in neocortical pyramidal cell dendrites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.01.494180v1?rss=1">
<title>
<![CDATA[
Male mice carrying the BDNF Val68Met polymorphism exhibit alcohol preferences versus social interaction and acute tolerance through malfunction of BDNF in the ventral hippocampus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.01.494180v1?rss=1</link>
<description><![CDATA[
The brain-derived neurotrophic factor (BDNF) Valine 66 to Methionine human polymorphism results in impaired activity-dependent BDNF release and has been linked to psychiatric disorders including depression and anxiety. We previously showed that male knock-in mice carrying the mouse Methionine homolog (Met68BDNF) exhibit excessive and compulsive alcohol drinking behaviors as compared to the wild-type Val68BDNF mice. Here, we set out to determine the potential mechanism for the heightened and compulsive alcohol drinking phenotypes detected in Met68BDNF mice. We found that male, but not female Met68BDNF mice exhibit social anxiety-like behaviors. We further show that male Met68BDNF mice exhibit a preference for alcohol over social interaction. In contrast, alcohol place preference without an alternative social reward, is similar in male Met68BDNF and Val68BDNF mice. Since the Met68BDNF mice show social anxiety phenotypes, we tested whether alcohol reliefs anxiety similarly in Met68BDNF and Val68BDNF mice and found that male, but not female Met68BDNF mice are insensitive to the acute anxiolytic action of alcohol. Finally, we show that this acute tolerance to alcohol-dependent anxiolysis can be restored by overexpressing wild-type Val68BDNF in the ventral hippocampus (vHC) of Met68BDNF mice. Together, our results suggest that excessive alcohol drinking in the Met68BDNF may be attributed, in part, to heighted social anxiety and a lack of alcohol-dependent anxiolysis, a phenotype that is associated with malfunction of BDNF signaling in the vHC of male Met68BDNF mice.
]]></description>
<dc:creator>Moffat, J. J.</dc:creator>
<dc:creator>Sakhai, S.</dc:creator>
<dc:creator>Hoisington, Z. W.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2022-06-03</dc:date>
<dc:identifier>doi:10.1101/2022.06.01.494180</dc:identifier>
<dc:title><![CDATA[Male mice carrying the BDNF Val68Met polymorphism exhibit alcohol preferences versus social interaction and acute tolerance through malfunction of BDNF in the ventral hippocampus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.06.495052v1?rss=1">
<title>
<![CDATA[
Quantitative proteomic analysis reveals apoE4-dependent phosphorylation of the actin-regulating protein VASP 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.06.495052v1?rss=1</link>
<description><![CDATA[
Apolipoprotein (apo) E4 is the major genetic risk factor for Alzheimers Disease (AD). While neurons generally produce a minority of the apoE in the central nervous system, neuronal expression of apoE increases dramatically in response to stress and is sufficient to drive pathology. Currently, the molecular mechanisms of how apoE4 expression may regulate pathology are not fully understood. Here we expand upon our previous studies measuring the impact of apoE4 on protein abundance to include the analysis of protein phosphorylation and ubiquitylation signaling in isogenic Neuro-2a cells expressing apoE3 or apoE4. ApoE4 expression resulted in a dramatic increase in VASP S235 phosphorylation in a PKA-dependent manner. This phosphorylation disrupted VASP interactions with numerous actin cytoskeletal and microtubular proteins. Reduction of VASP S235 phosphorylation via PKA inhibition resulted in a significant increase in filopodia formation and neurite outgrowth in apoE4-expressing cells, exceeding levels observed in apoE3-expressing cells. Our results highlight the pronounced and diverse impact of apoE4 on multiple modes of protein regulation and identify protein targets to restore apoE4-related cytoskeletal defects.
]]></description>
<dc:creator>Cakir, Z.</dc:creator>
<dc:creator>Lord, S. J.</dc:creator>
<dc:creator>Jimenez-Morales, D.</dc:creator>
<dc:creator>Jang, G. M.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Eckhardt, M.</dc:creator>
<dc:creator>Newton, B. W.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Orr, A. L.</dc:creator>
<dc:creator>Johnson, J. R.</dc:creator>
<dc:creator>Mullins, D.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Mahley, R. W.</dc:creator>
<dc:date>2022-06-07</dc:date>
<dc:identifier>doi:10.1101/2022.06.06.495052</dc:identifier>
<dc:title><![CDATA[Quantitative proteomic analysis reveals apoE4-dependent phosphorylation of the actin-regulating protein VASP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.10.495662v1?rss=1">
<title>
<![CDATA[
Mapping Protein Dynamics at High-Resolution with Temperature-Jump X-ray Crystallography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.10.495662v1?rss=1</link>
<description><![CDATA[
Understanding and controlling protein motion at atomic resolution is a hallmark challenge for structural biologists and protein engineers because conformational dynamics are essential for complex functions such as enzyme catalysis and allosteric regulation. Time-resolved crystallography offers a window into protein motions, yet without a universal perturbation to initiate conformational changes the method has been limited in scope. Here we couple a solvent-based temperature jump with time-resolved crystallography to visualize structural motions in lysozyme, a dynamic enzyme. We observed widespread atomic vibrations on the nanosecond timescale, which evolve on the sub-millisecond timescale into localized structural fluctuations that are coupled to the active site. An orthogonal perturbation to the enzyme, inhibitor binding, altered these dynamics by blocking key motions that allow energy to dissipate from vibrations into functional movements linked to the catalytic cycle. Because temperature-jump is a universal method for perturbing molecular motion, the method demonstrated here is broadly applicable for studying protein dynamics.
]]></description>
<dc:creator>Wolff, A. M.</dc:creator>
<dc:creator>Nango, E.</dc:creator>
<dc:creator>Young, I. D.</dc:creator>
<dc:creator>Brewster, A. S.</dc:creator>
<dc:creator>Kubo, M.</dc:creator>
<dc:creator>Nomura, T.</dc:creator>
<dc:creator>Sugahara, M.</dc:creator>
<dc:creator>Owada, S.</dc:creator>
<dc:creator>Barad, B. A.</dc:creator>
<dc:creator>Ito, K.</dc:creator>
<dc:creator>Bhowmick, A.</dc:creator>
<dc:creator>Carbajo, S.</dc:creator>
<dc:creator>Hino, T.</dc:creator>
<dc:creator>Holton, J. M.</dc:creator>
<dc:creator>Im, D.</dc:creator>
<dc:creator>O'Riordan, L. J.</dc:creator>
<dc:creator>Tanaka, T.</dc:creator>
<dc:creator>Tanaka, R.</dc:creator>
<dc:creator>Sierra, R. G.</dc:creator>
<dc:creator>Yumoto, F.</dc:creator>
<dc:creator>Tono, K.</dc:creator>
<dc:creator>Iwata, S.</dc:creator>
<dc:creator>Sauter, N. K.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Thompson, M. C.</dc:creator>
<dc:date>2022-06-12</dc:date>
<dc:identifier>doi:10.1101/2022.06.10.495662</dc:identifier>
<dc:title><![CDATA[Mapping Protein Dynamics at High-Resolution with Temperature-Jump X-ray Crystallography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.12.495827v1?rss=1">
<title>
<![CDATA[
A Development-Inspired Niche for Homeostatic Human Mini-Intestines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.12.495827v1?rss=1</link>
<description><![CDATA[
Epithelial organoids derived from intestinal tissue, also referred to as mini-intestines or mini-guts, recapitulate many aspects of the organ in vitro and can be used for biological discovery, personalized medicine, and drug development. Murine intestinal organoids represent a homeostatic system that balances stem cell maintenance within a crypt-like compartment and differentiation within a villus-like compartment1-3. However, this homeostatic balance and spatial organization has not been achieved with human intestinal organoids4. Here, we leverage single cell RNA-seq data (scRNA-seq) and high-resolution imaging to interrogate the developing human intestinal stem cell niche. We identified an EGF-family member, EPIREGULIN (EREG), as uniquely expressed in the developing crypt, and found that EREG can take the place of EGF as an in vitro niche factor. Unlike EGF, which leads to growth of thin-walled cystic organoids, EREG-organoids are spatially resolved into budded and proliferative crypt domains and a differentiated villus-like central lumen. Transcriptomics and epigenomics showed that EREG-organoids are globally similar to the native intestine while EGF-organoids have an altered chromatin landscape, downregulate the master intestinal transcription factor CDX25,6, and ectopically express stomach genes.
]]></description>
<dc:creator>Childs, C. J.</dc:creator>
<dc:creator>Holloway, E. M.</dc:creator>
<dc:creator>Sweet, C. W.</dc:creator>
<dc:creator>Tsai, Y.-H.</dc:creator>
<dc:creator>Wu, A.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Cardenas, O. P.</dc:creator>
<dc:creator>Capeling, M. M.</dc:creator>
<dc:creator>Eiken, M.</dc:creator>
<dc:creator>Zwick, R.</dc:creator>
<dc:creator>Palikuqi, B.</dc:creator>
<dc:creator>Trentesaux, C.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Glass, I.</dc:creator>
<dc:creator>Loebel, C.</dc:creator>
<dc:creator>Yu, Q.</dc:creator>
<dc:creator>Camp, G.</dc:creator>
<dc:creator>Sexton, J. Z.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Verzi, M. P.</dc:creator>
<dc:creator>Spence, J. J.</dc:creator>
<dc:date>2022-06-12</dc:date>
<dc:identifier>doi:10.1101/2022.06.12.495827</dc:identifier>
<dc:title><![CDATA[A Development-Inspired Niche for Homeostatic Human Mini-Intestines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.15.496340v1?rss=1">
<title>
<![CDATA[
Endocytic trafficking determines cellular tolerance of presynaptic opioid signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.15.496340v1?rss=1</link>
<description><![CDATA[
Opioid tolerance is well described physiologically but its mechanistic basis remains incompletely understood. An important site of opioid action in vivo is the presynaptic terminal, where opioids inhibit transmitter release. This response characteristically resists desensitization over minutes yet becomes gradually tolerant over hours, and how this is possible remains unknown. Here we delineate a cellular mechanism underlying this longer-term form of opioid tolerance. Our results support a model in which presynaptic tolerance is mediated by a gradual depletion of cognate receptors from the axon surface through iterative rounds of receptor endocytosis and recycling. For the -opioid receptor (MOR), we show that the agonist-induced endocytic process which initiates iterative receptor cycling requires GRK2/3-mediated phosphorylation of the receptors cytoplasmic tail, and that partial or biased agonist drugs which have reduced ability to drive phosphorylation-dependent endocytosis in terminals produce correspondingly less presynaptic tolerance. We then show that the {delta}-opioid receptor (DOR) conforms to the same general paradigm except that endocytosis of DOR from the presynapse, in marked contrast to MOR, does not require phosphorylation of the receptors cytoplasmic tail. Further, we show that DOR recycles less efficiently than MOR in axons and, consistent with this, that DOR tolerance develops more strongly. Together, these results delineate a cellular basis for the development of presynaptic tolerance to opioids and describe a methodology useful for investigating presynaptic neuromodulation more broadly.
]]></description>
<dc:creator>Jullie, D.</dc:creator>
<dc:creator>Benitez, C.</dc:creator>
<dc:creator>Knight, T. A.</dc:creator>
<dc:creator>Von Zastrow, M.</dc:creator>
<dc:date>2022-06-16</dc:date>
<dc:identifier>doi:10.1101/2022.06.15.496340</dc:identifier>
<dc:title><![CDATA[Endocytic trafficking determines cellular tolerance of presynaptic opioid signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.16.496371v1?rss=1">
<title>
<![CDATA[
Early and lifelong effects of APOE4 on neuronal gene expression networks relevant to Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.16.496371v1?rss=1</link>
<description><![CDATA[
Apolipoprotein E4 (APOE4) genotype and aging are critical risk factors for Alzheimers disease (AD). Aged APOE4 knock-in (APOE4-KI) mice have phenotypes reflecting features of AD. We conducted a large-scale single nucleus RNA-sequencing study to identify cell-type-specific effects of APOE4 on hippocampal gene expression during aging. APOE4-KI mice showed prominent alterations, relative to APOE3-KI mice, in neuronal transcriptome related to synaptic function, calcium signaling, and MAPK/Rap1/Pld signal transduction, starting by 5 months and persisting during aging. Mice with the APOE4 gene removed specifically from neurons failed to show most of these neuronal transcriptomic changes, suggesting a specific effect of neuron-derived APOE4 on the transcriptome. APOE4 affects similar cellular pathways in induced pluripotent stem cell-derived human neurons transplanted into APOE4-KI mouse hippocampus and in cortical neurons from aged human brains. Thus, neuronal APOE4 has early and persistent effects on neuronal transcriptomes, suggesting the requirement of early interventions for successfully treating APOE4-related AD.
]]></description>
<dc:creator>Grone, B.</dc:creator>
<dc:creator>Zalocusky, K.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Yoon, S. Y.</dc:creator>
<dc:creator>Arriola, P.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:date>2022-06-17</dc:date>
<dc:identifier>doi:10.1101/2022.06.16.496371</dc:identifier>
<dc:title><![CDATA[Early and lifelong effects of APOE4 on neuronal gene expression networks relevant to Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.16.496455v1?rss=1">
<title>
<![CDATA[
Dynamic top-down biasing implements rapid adaptive changes to individual movements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.16.496455v1?rss=1</link>
<description><![CDATA[
Complex behaviors depend on the coordinated activity of neural ensembles in interconnected brain areas. The behavioral function of such coordination, often measured as co-fluctuations in neural activity across areas, is poorly understood. One hypothesis is that rapidly varying co-fluctuations may be a signature of moment-by-moment task-relevant influences of one area on another. We tested this possibility for error-corrective adaptation of birdsong, a form of motor learning which has been hypothesized to depend on the top-down influence of a higher-order area, LMAN, in shaping moment-by-moment output from a primary motor area, RA. In paired recordings of LMAN and RA in singing birds, we discovered a neural signature of a top-down influence of LMAN on RA, quantified as an LMAN-leading co-fluctuation between these areas. During learning, this cofluctuation strengthened in a premotor temporal window linked to the specific movement, sequential context, and acoustic modification associated with learning. Moreover, transient perturbation of LMAN activity specifically within this premotor window caused rapid occlusion of pitch modifications, consistent with LMAN conveying a temporally localized motor-biasing signal. Combined, our results reveal a dynamic top-down influence of LMAN on RA that varies on the rapid timescale of individual movements and is flexibly linked to contexts associated with learning. This finding indicates that inter-area co-fluctuations can be a signature of dynamic top-down influences that support complex behavior and its adaptation.
]]></description>
<dc:creator>Tian, L. Y.</dc:creator>
<dc:creator>Warren, T. L.</dc:creator>
<dc:creator>Brainard, M. S.</dc:creator>
<dc:date>2022-06-17</dc:date>
<dc:identifier>doi:10.1101/2022.06.16.496455</dc:identifier>
<dc:title><![CDATA[Dynamic top-down biasing implements rapid adaptive changes to individual movements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.23.496464v1?rss=1">
<title>
<![CDATA[
Systems Analysis of de novo Mutations in Congenital Heart Diseases Identified a Molecular Network in Hypoplastic Left Heart Syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.23.496464v1?rss=1</link>
<description><![CDATA[
Congenital heart diseases (CHD) are a class of birth defects affecting [~]1% of live births. These conditions are hallmarked by extreme genetic heterogeneity, and therefore, despite a strong genetic component, only a very handful of at-risk loci in CHD have been identified. We herein introduced systems analyses to uncover the hidden organization on biological networks of genomic mutations in CHD, and leveraged network analysis techniques to integrate the human interactome, large-scale patient exomes, the fetal heart spatial transcriptomes, and single-cell transcriptomes of clinical samples. We identified a highly connected network in CHD where most of the member proteins had previously uncharacterized functions in regulating fetal heart development. While genes on the network displayed strong enrichment for heart-specific functions, a sub-group, active specifically at early developmental stages, also regulates fetal brain development, thereby providing mechanistic insights into the clinical comorbidities between CHD and neurodevelopmental conditions. At a small scale, we experimentally verified previously uncharacterized cardiac functions of several novel proteins employing cellular assays and gene editing techniques. At a global scale, our study revealed developmental dynamics of the identified CHD network and observed the strongest enrichment for pathogenic mutations in the network specific to hypoplastic left heart syndrome (HLHS). Our single-cell transcriptome analysis further identified pervasive dysregulation of the network in cardiac endothelial cells and the conduction system in the HLHS left ventricle. Taken together, our systems analyses identified novel factors in CHD, revealed key molecular mechanisms in HLHS, and provides a generalizable framework readily applicable to studying many other complex diseases.
]]></description>
<dc:creator>Wang, Y. J.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Lam, C. K.</dc:creator>
<dc:creator>Guo, H.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Wu, J. C.</dc:creator>
<dc:creator>Snyder, M.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:date>2022-06-26</dc:date>
<dc:identifier>doi:10.1101/2022.06.23.496464</dc:identifier>
<dc:title><![CDATA[Systems Analysis of de novo Mutations in Congenital Heart Diseases Identified a Molecular Network in Hypoplastic Left Heart Syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.30.498336v1?rss=1">
<title>
<![CDATA[
Pitfalls of genotyping microbial communities with rapidly growing genome collections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.30.498336v1?rss=1</link>
<description><![CDATA[
Detecting genetic variants in metagenomic data is a priority for understanding the evolution, ecology, and functional characteristics of microbial communities. Many recent tools that perform this metagenotyping rely on aligning reads of unknown origin to a reference database of sequences from many species before calling variants. Using simulations designed to represent a wide range of scenarios, we demonstrate that diverse and closely related species both reduce the power and accuracy of reference-based metagenotyping. We identify multi-mapping reads as a prevalent source of errors and illustrate a tradeoff between retaining correct alignments versus limiting incorrect alignments, many of which map reads to the wrong species. Then we quantitatively evaluate several actionable mitigation strategies and review emerging methods with promise to further improve metagenotyping. These findings document a critical challenge that has come to light through the rapid growth of genome collections that push the limits of current alignment algorithms. Our results have implications beyond metagenotyping to the many tools in microbial genomics that depend upon accurate read mapping.

HIGHLIGHTSO_LIMost microbial species are genetically diverse. Their single nucleotide variants can be genotyped using metagenomic data aligned to databases constructed from genome collections ("metagenotyping").
C_LIO_LIMicrobial genome collections have grown and now contain many pairs of closely related species.
C_LIO_LIClosely related species produce high-scoring but incorrect alignments while also reducing the uniqueness of correct alignments. Both cause metagenotype errors.
C_LIO_LIThis dilemma can be mitigated by leveraging paired-end reads, customizing databases to species detected in the sample, and adjusting post-alignment filters.
C_LI
]]></description>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Shi, Z. J.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2022-07-02</dc:date>
<dc:identifier>doi:10.1101/2022.06.30.498336</dc:identifier>
<dc:title><![CDATA[Pitfalls of genotyping microbial communities with rapidly growing genome collections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.02.498578v1?rss=1">
<title>
<![CDATA[
Membrane curvature sensing and symmetry breaking of the M2 proton channel from Influenza A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.02.498578v1?rss=1</link>
<description><![CDATA[
The M2 proton channel aids in the exit of mature influenza viral particles from the host plasma membrane through its ability to stabilize regions of high negative gaussian curvature (NGC) that occur at the neck of budding virions. The channels are homo-tetramers that contain a cytoplasm-facing amphipathic helix (AH) that is necessary and sufficient for NGC generation; however, constructs containing the transmembrane spanning helix, which facilitates tetramerization, exhibit enhanced curvature generation. Here we used all-atom molecular dynamics (MD) simulations to explore the conformational dynamics of M2 channels in lipid bilayers revealing that the AH is dynamic, quickly breaking the 4-fold symmetry observed in most structures. Next, we carried out MD simulations with the protein restrained in 4-fold and 2-fold symmetric conformations to determine the impact on the membrane shape. While each pattern was distinct, all configurations induced pronounced curvature in the outer leaflet with rather subtle lipid tilt, while conversely, the inner leaflets adjacent to the AHs showed minimal curvature and significant lipid tilt. The MD-generated profiles at the protein-membrane interface were then extracted and used as boundary conditions in a continuum elastic membrane model to calculate the membrane bending energy of each conformation embedded in different membrane surfaces characteristic of a budding virus. The calculations show that all three M2 conformations are stabilized in concave spherical caps and destabilized in convex spherical caps, the latter reminiscent of a budding virus. Only C2-broken symmetry conformations are stabilized in NGC surfaces, by 1-3 kBT depending on the AH domain arrangement. The most favored conformation is stabilized in saddles with curvatures corresponding to 33 nm radii. In total, our work provides atomistic insight into the curvature sensing capabilities of M2 channels and how enrichment in the nascent viral particle depends on protein shape and membrane geometry.
]]></description>
<dc:creator>Helsell, C. V. M.</dc:creator>
<dc:creator>Marcoline, F. V.</dc:creator>
<dc:creator>Lincoff, J.</dc:creator>
<dc:creator>Natale, A. M.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:date>2022-07-05</dc:date>
<dc:identifier>doi:10.1101/2022.07.02.498578</dc:identifier>
<dc:title><![CDATA[Membrane curvature sensing and symmetry breaking of the M2 proton channel from Influenza A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.13.499965v1?rss=1">
<title>
<![CDATA[
GPCR-dependent spatiotemporal cAMP generation confers functional specificity in cardiomyocytes and cardiac responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.13.499965v1?rss=1</link>
<description><![CDATA[
Cells interpret a variety of signals through G protein-coupled receptors (GPCRs) and stimulate the generation of second messengers such as cyclic adenosine monophosphate (cAMP). A long-standing puzzle is deciphering how GPCRs elicit different physiological responses despite generating similar levels of cAMP. We previously showed that some GPCRs generate cAMP from both the plasma membrane and the Golgi apparatus. Here, we demonstrate that cardiomyocytes distinguish between subcellular cAMP inputs to elicit different physiological outputs. We show that generating cAMP from the Golgi leads to regulation of a specific PKA target that increases the rate of cardiomyocyte relaxation. In contrast, cAMP generation from the plasma membrane activates a different PKA target that increases contractile force. We further validated the physiological consequences of these observations in intact zebrafish and mice. Thus, we demonstrate that the same GPCR acting through the same second messenger regulates cardiac contraction and relaxation dependent on its subcellular location.
]]></description>
<dc:creator>Irannejad, R.</dc:creator>
<dc:creator>Mai, Q. N.</dc:creator>
<dc:creator>Lin, T.-Y.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Wilson, E.</dc:creator>
<dc:creator>Chien, H.-C.</dc:creator>
<dc:creator>Yee, S. W.</dc:creator>
<dc:creator>Giacomini, K. M.</dc:creator>
<dc:creator>Olgin, J. E.</dc:creator>
<dc:date>2022-07-13</dc:date>
<dc:identifier>doi:10.1101/2022.07.13.499965</dc:identifier>
<dc:title><![CDATA[GPCR-dependent spatiotemporal cAMP generation confers functional specificity in cardiomyocytes and cardiac responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.15.500120v1?rss=1">
<title>
<![CDATA[
Systematic functional interrogation of SARS-CoV-2 host factors using Perturb-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.15.500120v1?rss=1</link>
<description><![CDATA[
Genomic and proteomic screens have identified numerous host factors of SARS-CoV-2, but efficient delineation of their molecular roles during infection remains a challenge. Here we use Perturb-seq, combining genetic perturbations with a single-cell readout, to investigate how inactivation of host factors changes the course of SARS-CoV-2 infection and the host response in human lung epithelial cells. Our high-dimensional data resolve complex phenotypes such as shifts in the stages of infection and modulations of the interferon response. However, only a small percentage of host factors showed such phenotypes upon perturbation. We further identified the NF-{kappa}B inhibitor I{kappa}B (NFKBIA), as well as the translation factors EIF4E2 and EIF4H as strong host dependency factors acting early in infection. Overall, our study provides massively parallel functional characterization of host factors of SARS-CoV-2 and quantitatively defines their roles both in virus-infected and bystander cells.
]]></description>
<dc:creator>Sunshine, S.</dc:creator>
<dc:creator>Puschnik, A. S.</dc:creator>
<dc:creator>Replogle, J. M.</dc:creator>
<dc:creator>Laurie, M. T.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Zha, B. S.</dc:creator>
<dc:creator>Nunez, J. K.</dc:creator>
<dc:creator>Byrum, J. R.</dc:creator>
<dc:creator>McMorrow, A. H.</dc:creator>
<dc:creator>Frieman, M. B.</dc:creator>
<dc:creator>Winkler, J.</dc:creator>
<dc:creator>Qiu, X.</dc:creator>
<dc:creator>Rosenberg, O. S.</dc:creator>
<dc:creator>Leonetti, M. D.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Hein, M. Y.</dc:creator>
<dc:date>2022-07-17</dc:date>
<dc:identifier>doi:10.1101/2022.07.15.500120</dc:identifier>
<dc:title><![CDATA[Systematic functional interrogation of SARS-CoV-2 host factors using Perturb-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.18.500281v1?rss=1">
<title>
<![CDATA[
Evidence for intercellular bridges and radial patterning of meiotic initiation in the human fetal ovary 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.18.500281v1?rss=1</link>
<description><![CDATA[
Meiosis is the hallmark of reproduction. Our understanding of early oocyte development was improved by studying the spatiotemporal dynamics and mechanisms governing meiosis in mice, however, our knowledge of the meiotic initiation process in humans remains limited. Here, we utilized three-dimensional (3D) analysis to determine spatiotemporal dynamics of meiotic initiation in fetal human ovaries. Similar to mice, we found that the first meiotic cells appear in clusters in the center of human fetal ovaries as early at 9 weeks and that the initiation of meiosis propagates as a radial wave. Between developing germ cells in fetal human ovaries, we detected a component of the intercellular bridge, TEX14 protein. 3D quantification of germ cells in ovaries collected at the end of first trimester revealed, for the first time, considerable variation in the number of meiotic cells between individuals and asynchronous mitotic-meiotic transition. In addition to illustrating the geography of meiotic initiation in ovaries from the first trimester, we extended our 3D analysis approach to second trimester and showed heterogeneous spatial distribution of meiotic and non-meiotic germ cells in human fetal ovaries. This study is an important step towards better understanding of 3D structure of developing ovary and early stages of meiosis in humans.

Highlights- Organic solvent-based clearing methods and confocal microscopy can be implemented to visualize and quantify germ cells in the intact human fetal ovary.
- Identification of a new, radial, pattern of meiotic initiation in the ovaries from the first trimester.
- Immunolocalization of the intercellular bridge component TEX14 between developing germ cells in the fetal ovary
- Maintenance of pre-meiotic germ cells in second trimester ovaries suggests less synchronous mitotic-meiotic stage transition.
]]></description>
<dc:creator>Soygur, B.</dc:creator>
<dc:creator>Derpinghaus, A.</dc:creator>
<dc:creator>Cunha, G. R.</dc:creator>
<dc:creator>Baskin, L. S.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:date>2022-07-18</dc:date>
<dc:identifier>doi:10.1101/2022.07.18.500281</dc:identifier>
<dc:title><![CDATA[Evidence for intercellular bridges and radial patterning of meiotic initiation in the human fetal ovary]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.23.501237v1?rss=1">
<title>
<![CDATA[
Wireless Measurements Using Electrical Impedance Spectroscopy to Monitor Fracture Healing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.23.501237v1?rss=1</link>
<description><![CDATA[
There is an unmet need for improved, clinically relevant methods to longitudinally quantify bone healing during fracture care. Here we develop a smart bone plate to wirelessly monitor healing utilizing electrical impedance spectroscopy (EIS) to provide real-time data on tissue com-position within the fracture callus. To validate our technology, we created a 1-mm rabbit tibial defect and fixed the bone with a standard veterinary plate modified with a custom-designed housing that included two impedance sensors capable of wireless transmission. Impedance magnitude and phase measurements were transmitted every 48-hours for up to 10-weeks. Bone healing was assessed by X-ray, CT, and histomorphometry. Our results indicated the sensors successfully incorporated into the fracture callus and did not impede repair. Electrical impedance, resistance, and reactance increased steadily from weeks 3 to 7--corresponding to the transition from hematoma to cartilage to bone within the fracture gap--then plateaued as the bone began to consolidate. These three electrical readings significantly correlated with traditional measurements of bone healing and successfully distinguished between union and not healed fractures, with the strongest relationship found with impedance magnitude. These results suggest that our EIS smart bone plate can provide continuous and highly sensitive quantitative tissue measurements throughout the course of fracture healing to better guide personalized clinical care.
]]></description>
<dc:creator>Fukase, N.</dc:creator>
<dc:creator>Duke, V. R.</dc:creator>
<dc:creator>Lin, M. C.</dc:creator>
<dc:creator>Stake, I. K.</dc:creator>
<dc:creator>Huard, M.</dc:creator>
<dc:creator>Huard, J.</dc:creator>
<dc:creator>Marmor, M. T.</dc:creator>
<dc:creator>Maharbiz, M. M.</dc:creator>
<dc:creator>Ehrhart, N. P.</dc:creator>
<dc:creator>Bahney, C. S.</dc:creator>
<dc:creator>Herfat, S. T.</dc:creator>
<dc:date>2022-07-23</dc:date>
<dc:identifier>doi:10.1101/2022.07.23.501237</dc:identifier>
<dc:title><![CDATA[Wireless Measurements Using Electrical Impedance Spectroscopy to Monitor Fracture Healing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.25.501254v1?rss=1">
<title>
<![CDATA[
Neuronal Activity Alters Neuron to OPC Synapses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.25.501254v1?rss=1</link>
<description><![CDATA[
The mechanisms that drive the timing and specificity of oligodendrocyte myelination during development, or remyelination after injury or immune attack are not well understood. Recent work has shown that oligodendrocyte progenitors receive synapses from neurons, providing a potential mechanism for neuronal-glial communication. We hypothesize that these connections are important both for correct myelination of neurons during development and for myelination during neuronal plasticity. We utilized chemogenetic tools and viral monosynaptic circuit tracing to analyze these neuroglial connections and to examine OPC proliferation, myelination, synapse formation, and neuronal-glial connectivity after increasing or decreasing neuronal activity in vivo. We found that increasing neuronal activity increased OPC activation, but not proliferation. We also found that altering neuronal activity altered neuronal-glial synaptic connections: while it did not impact the total number of neuronal inputs, or the number of inhibitory neuronal inputs, it did alter the number of excitatory neuron to OPC connections. We also found that increasing or decreasing neuronal activity impacted the ratio of excitatory and inhibitory synapses. Our data show that neuronal activity affects OPC activation, neuronal synapse formation onto OPCs, as well as the types of neuronal inputs to OPCs, indicating that neuronal activity is important for OPC circuit composition and function.
]]></description>
<dc:creator>Moura, D. S.</dc:creator>
<dc:creator>Parvathaneni, A.</dc:creator>
<dc:creator>Sahagun, A.</dc:creator>
<dc:creator>Noguchi, H.</dc:creator>
<dc:creator>Tilton, I.</dc:creator>
<dc:creator>Brennan, E.</dc:creator>
<dc:creator>Brock, R. M.</dc:creator>
<dc:creator>Halladay, L. R.</dc:creator>
<dc:creator>Pleasure, S. J.</dc:creator>
<dc:creator>Cocas, L. A.</dc:creator>
<dc:date>2022-07-25</dc:date>
<dc:identifier>doi:10.1101/2022.07.25.501254</dc:identifier>
<dc:title><![CDATA[Neuronal Activity Alters Neuron to OPC Synapses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.26.501550v1?rss=1">
<title>
<![CDATA[
Allosteric Regulation of a Synaptic Vesicle Glutamate Transporter 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.26.501550v1?rss=1</link>
<description><![CDATA[
Concentration of neurotransmitter inside synaptic vesicles (SVs) underlies the quantal nature of synaptic transmission. In contrast to many transporters, SV uptake of the principal excitatory neurotransmitter glutamate is driven by membrane potential. To prevent nonquantal efflux of glutamate after SV exocytosis, the vesicular glutamate transporters (VGLUTs) are allosterically inhibited by the neutral pH of the synaptic cleft. We have now determined high-resolution structures of VGLUT2 with a cyclic analog of glutamate bound that defines the mechanism of substrate recognition, a positively charged cytoplasmic vestibule that electrostatically attracts the negatively charged substrate, and modification by palmitoylation that promotes retrieval of the transporter after exocytosis. The structure also incorporates an extensive, cytoplasmic network of electrostatic interactions that acts as a gate. Functional analysis shows how this cytoplasmic gate confers the allosteric requirement for lumenal H+ required to restrict VGLUT activity to SVs.
]]></description>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Eriksen, J.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>Gomez, Y. K.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Finer-Moore, J.</dc:creator>
<dc:creator>Nguyen, P.</dc:creator>
<dc:creator>Bowen, A.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Burlingame, A.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>Stroud, R.</dc:creator>
<dc:creator>Edwards, R.</dc:creator>
<dc:date>2022-07-26</dc:date>
<dc:identifier>doi:10.1101/2022.07.26.501550</dc:identifier>
<dc:title><![CDATA[Allosteric Regulation of a Synaptic Vesicle Glutamate Transporter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.27.501186v1?rss=1">
<title>
<![CDATA[
Modular Pooled Discovery of Synthetic Knockin Sequences to Program Durable Cell Therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.27.501186v1?rss=1</link>
<description><![CDATA[
Chronic stimulation can cause T cell dysfunction and limit efficacy of cellular immunotherapies. CRISPR screens have nominated gene targets for engineered T cells, but improved methods are required to compare large numbers of synthetic knockin sequences to reprogram cell functions. Here, we developed Modular Pooled Knockin Screening (ModPoKI), an adaptable platform for modular construction of DNA knockin libraries using barcoded multicistronic adaptors. We built two ModPoKI libraries of 100 transcription factors (TFs) and 129 natural and synthetic surface receptors. Over 20 ModPoKI screens across human TCR and CAR T cells in diverse conditions identified a transcription factor AP4 (TFAP4) construct to enhance long-term T cell fitness and anti-cancer function in vitro and in vivo. ModPoKIs modularity allowed us to generate a [~]10,000-member library of TF combinations. Non-viral knockin of a combined BATF-TFAP4 polycistronic construct further enhanced function in vivo. ModPoKI facilitates discovery of complex gene constructs to program cellular functions.

HighlightsO_LIModular pooled knockins of hundreds of TF and surface receptor constructs combined with different antigen receptors
C_LIO_LIChronic stimulation screens discover programs to improve T cell persistence
C_LIO_LICombinatorial knockin screens with [~]10,000 transcription factor combinations
C_LIO_LIBATF-TFAP4 dual knockin construct improves CAR T cell function in vitro and in vivo
C_LI
]]></description>
<dc:creator>Blaeschke, F.</dc:creator>
<dc:creator>Chen, Y. Y.</dc:creator>
<dc:creator>Apathy, R.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Mowery, C. T.</dc:creator>
<dc:creator>Nyberg, W. A.</dc:creator>
<dc:creator>To, A.</dc:creator>
<dc:creator>Yu, R.</dc:creator>
<dc:creator>Bueno, R.</dc:creator>
<dc:creator>Kim, M. C.</dc:creator>
<dc:creator>Schmidt, R.</dc:creator>
<dc:creator>Goodman, D. B.</dc:creator>
<dc:creator>Feuchtinger, T.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Roth, T. L.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2022-07-27</dc:date>
<dc:identifier>doi:10.1101/2022.07.27.501186</dc:identifier>
<dc:title><![CDATA[Modular Pooled Discovery of Synthetic Knockin Sequences to Program Durable Cell Therapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.30.502150v1?rss=1">
<title>
<![CDATA[
Differential Analysis of Stromal-Epithelial Interactions between In Situ and Invasive Breast Cancer using Gene Expression Profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.30.502150v1?rss=1</link>
<description><![CDATA[
BackgroundChanges in microenvironment cell-cell interactions (CCI) during the progression from ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) are poorly understood. Gene expression studies are confounded by cellular heterogeneity and few separate stromal and epithelial contributions, resulting in a lack of reliable prognostic biomarker to guide treatment decisions.

MethodsThe gene expression of 293 microdissected regions from DCIS (92 epithelial, 31 stromal) and IDC (78 epithelial, 30 stromal) cases was aggregated from 6 datasets. Expression signatures of 6 cell lineages extracted from normal breast single-cell profiling were used to correct for differences in cell abundance. Subtype-specific functional differences between DCIS and IDC were measured for each region type using Gene Set Enrichment Analysis (GSEA). DCIS-IDC stromal-epithelial interactions were compared using the expression product of 139 ligand-receptor (LR) pairs permuting the DCIS-IDC labels to assess significance.

ResultsVariation in cell-lineage abundance separated epithelial regions into 4 clusters, including one enriched for DCIS (Myoepi-Enriched) and two for IDC (Infiltrated, Vascularized). GSEA on cell lineage normalized expression data identified subtype-independent changes in epithelial regions (induction of Extracellular Matrix maintenance genes, reduction of Tp53 signaling in IDC), as well as subtype-specific changes (proliferation in ER- and Her2-IDC, reduction in Nucleotide Excision Repair in ER+ IDC). In the stroma, Notch and Rho-GTPase signaling were induced in IDC irrespective of subtype. The stromal-epithelial interaction level of 6 and 4 LR pairs were significantly enriched in DCIS and IDC, respectively. Five of the 6 DCIS-enriched LR pairs involved ephrin interactions, with interaction level progressively decreasing from normal to DCIS to IDC. In contrast, 2 IDC-enriched LR pairs involved T-cell activity likely regulating Treg proliferation (CD28-CD86) or T and NK cells stimulation (CD226-PVR). Notably, the bulk expression product of one identified LR pair (EPHB4-EFNB1) was associated with poor survival in IDC (HR=1.47, p=0.04) suggesting that early remodeling of this stromal-epithelial interaction may have long-lasting impact on disease severity.

ConclusionsThe observed changes in cell states and stromal-epithelial interactions, beyond those driven by difference in cell abundance, may lead to new biomarkers for prognosis and targets for secondary prevention.
]]></description>
<dc:creator>Officer, A.</dc:creator>
<dc:creator>Dempsey, A. M.</dc:creator>
<dc:creator>Murrow, L. M.</dc:creator>
<dc:creator>Gartner, Z.</dc:creator>
<dc:creator>Tamayo, P.</dc:creator>
<dc:creator>Yau, C.</dc:creator>
<dc:creator>Harismendy, O.</dc:creator>
<dc:date>2022-08-02</dc:date>
<dc:identifier>doi:10.1101/2022.07.30.502150</dc:identifier>
<dc:title><![CDATA[Differential Analysis of Stromal-Epithelial Interactions between In Situ and Invasive Breast Cancer using Gene Expression Profiling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.01.502349v1?rss=1">
<title>
<![CDATA[
Profile of the somatic mutational landscape in breast tumors from Hispanic/Latina women 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.01.502349v1?rss=1</link>
<description><![CDATA[
Breast cancer causes the most cancer deaths among Hispanic/Latinas (H/L). However, limited tumor-sequencing data from H/L are available to guide treatment. To address this gap, we performed whole-exome sequencing of DNA from 140 HL germline and 146 matched breast tumors and RNA-seq for the tumors. We generated somatic-mutation profiles, identified copy-number alterations (CNAs), and compared results to non-Hispanic White (White) women in The Cancer Genome Atlas. Similar to Whites, PIK3CA and TP53 were the most commonly mutated genes in breast tumors from H/L. We found 4 common COSMIC mutation signatures (1, 2, 3, 13) and signature 16 not previously reported in other breast-cancer datasets. We observed recurrent amplifications in breast-cancer drivers including MYC, FGFR1, CCND1, and ERBB2, and a recurrent amplification on 17q11.2 associated with high KIAA0100 gene expression, implicated in breast-cancer aggressiveness. Expanded research is required to determine how these characteristics of H/L tumors impact treatment response and survival.
]]></description>
<dc:creator>Ding, Y. C.</dc:creator>
<dc:creator>Song, H.</dc:creator>
<dc:creator>Adamson, A.</dc:creator>
<dc:creator>Schmolze, D.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Huntsman, S.</dc:creator>
<dc:creator>Steele, L.</dc:creator>
<dc:creator>Patrick, C.</dc:creator>
<dc:creator>Hernandez, N.</dc:creator>
<dc:creator>Adams, C. D.</dc:creator>
<dc:creator>Fejerman, L.</dc:creator>
<dc:creator>Gardner, K. L.</dc:creator>
<dc:creator>Napoles, A. M.</dc:creator>
<dc:creator>Perez-Stable, E. J.</dc:creator>
<dc:creator>Weitzel, J. N.</dc:creator>
<dc:creator>Bengtsson, H.</dc:creator>
<dc:creator>Huang, F. W.</dc:creator>
<dc:creator>Neuhausen, S. L.</dc:creator>
<dc:creator>Ziv, E.</dc:creator>
<dc:date>2022-08-03</dc:date>
<dc:identifier>doi:10.1101/2022.08.01.502349</dc:identifier>
<dc:title><![CDATA[Profile of the somatic mutational landscape in breast tumors from Hispanic/Latina women]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.03.502739v1?rss=1">
<title>
<![CDATA[
Optogenetic control of RelA reveals effect of transcription factor dynamics on downstream gene expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.03.502739v1?rss=1</link>
<description><![CDATA[
Many transcription factors (TFs) translocate to the nucleus with varied dynamic patterns in response to different inputs. A notable example of such behavior is RelA, a subunit of NF-{kappa}B, which translocates to the nucleus with either pulsed or sustained dynamics, depending on the stimulus. Our understanding of how these dynamics are interpreted by downstream genes has remained incomplete, partly because ubiquitously used environmental inputs activate other transcriptional regulators in addition to RelA. Here, we use an optogenetic tool, CLASP (controllable light-activated shuttling and plasma membrane sequestration), to control RelA spatiotemporal dynamics in mouse fibroblasts and quantify their effect on downstream genes using RNA-seq. Using RelA-CLASP, we show for the first time that nuclear translocation of RelA, without post-translational modifications or activation of other transcriptional regulators, is sufficient to activate downstream genes. Furthermore, we find that TNF, a common endogenous input, regulates many genes independently of RelA, and that this gene regulation is different from that induced by RelA-CLASP. Genes responsive to RelA-CLASP show a wide range of dynamics in response to a constant RelA input. We use a simple promoter model to recapitulate these diverse dynamic responses, as well as data collected in response to a pulsed RelA-CLASP input, and extract features of many RelA-responsive promoters. We also pinpoint many genes for which more complex models, involving feedback or multi-step promoters, may be needed to explain their response to constant and pulsed TF inputs. This study introduces a new robust tool for studying mammalian transcriptional regulation and demonstrates the power of optogenetic tools in dissecting the quantitative features of important cellular pathways.
]]></description>
<dc:creator>Osimiri, L. C.</dc:creator>
<dc:creator>Bonny, A. R.</dc:creator>
<dc:creator>Takagishi, S. R.</dc:creator>
<dc:creator>Luecke, S.</dc:creator>
<dc:creator>Riehs, N.</dc:creator>
<dc:creator>Hoffmann, A.</dc:creator>
<dc:creator>El-Samad, H.</dc:creator>
<dc:date>2022-08-05</dc:date>
<dc:identifier>doi:10.1101/2022.08.03.502739</dc:identifier>
<dc:title><![CDATA[Optogenetic control of RelA reveals effect of transcription factor dynamics on downstream gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.07.502893v1?rss=1">
<title>
<![CDATA[
Sensitive multimodal profiling of native DNA by transposase-mediated single-molecule sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.07.502893v1?rss=1</link>
<description><![CDATA[
We present SMRT-Tag: a multiplexable, PCR-free approach for constructing low-input, single-molecule Pacific Biosciences (PacBio) sequencing libraries through Tn5 transposition. As proof-of-concept, we apply SMRT-Tag to resolve human genetic and epigenetic variation in gold-standard human reference samples. SMRT-Tag requires 1-5% as much input material as existing protocols (15,000 - 50,000 human cell equivalents) and enables highly-sensitive and simultaneous detection of single nucleotide variants, small insertions / deletions, and CpG methylation comparable to the current state-of-the-art. We further combine SMRT-Tag with in situ adenine methyltransferase footprinting of nuclei (SAMOSA-Tag) to facilitate joint analysis of nucleosome repeat length, CTCF occupancy, and CpG methylation on individual chromatin fibers in osteosarcoma cells. SMRT-Tag promises to enable basic and clinical research by offering scalable, sensitive, and multimodal single-molecule genomic and epigenomic analyses in rare cell populations.
]]></description>
<dc:creator>Nanda, A. S.</dc:creator>
<dc:creator>Wu, K.</dc:creator>
<dc:creator>Kasinathan, S.</dc:creator>
<dc:creator>Ostrowski, M. S.</dc:creator>
<dc:creator>Clugston, A. S.</dc:creator>
<dc:creator>Satpathy, A.</dc:creator>
<dc:creator>Sweet-Cordero, E. A.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:date>2022-08-07</dc:date>
<dc:identifier>doi:10.1101/2022.08.07.502893</dc:identifier>
<dc:title><![CDATA[Sensitive multimodal profiling of native DNA by transposase-mediated single-molecule sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.14.503891v1?rss=1">
<title>
<![CDATA[
Reliable energy-based antibody humanization and stabilization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.14.503891v1?rss=1</link>
<description><![CDATA[
Humanization is an essential step in developing animal-derived antibodies into therapeutics, and approximately one third of approved antibodies have been humanized. Conventional humanization approaches graft the complementarity-determining regions (CDRs) of the animal antibody onto several homologous human frameworks. This process, however, often drastically lowers stability and antigen binding, demanding iterative mutational fine-tuning to recover the original antibody properties. Here, we present Computational hUMan AntiBody design (CUMAb), a web-accessible method that starts from an experimental or model antibody structure, systematically grafts the animal CDRs on thousands of human frameworks, and uses Rosetta atomistic simulations to rank the designs by energy and structural integrity (http://CUMAb.weizmann.ac.il). CUMAb designs of five independent antibodies exhibit similar affinities to the parental animal antibody, and some designs show marked improvement in stability. Surprisingly, nonhomologous frameworks are often preferred to the highest-homology ones, and several CUMAb designs that use different human frameworks and differ by dozens of mutations are functionally equivalent. Thus, CUMAb presents a general and streamlined approach to optimizing antibody stability and expressibility while increasing humanness.
]]></description>
<dc:creator>Tennenhouse, A.</dc:creator>
<dc:creator>Khmelnitsky, L.</dc:creator>
<dc:creator>Yeshaya, N.</dc:creator>
<dc:creator>Noronha, A.</dc:creator>
<dc:creator>Lindzen, M.</dc:creator>
<dc:creator>Khalaila, R.</dc:creator>
<dc:creator>Zaretsky, I.</dc:creator>
<dc:creator>Fridmann Sirkis, Y.</dc:creator>
<dc:creator>Galon-Wolfenson, Y.</dc:creator>
<dc:creator>Abramson, J.</dc:creator>
<dc:creator>Yarden, Y.</dc:creator>
<dc:creator>Fass, D.</dc:creator>
<dc:creator>Fleishman, S. J.</dc:creator>
<dc:date>2022-08-15</dc:date>
<dc:identifier>doi:10.1101/2022.08.14.503891</dc:identifier>
<dc:title><![CDATA[Reliable energy-based antibody humanization and stabilization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.15.504025v1?rss=1">
<title>
<![CDATA[
Single-cell RNA sequencing identifies regulators of differentiation and nutritional cues in Drosophila female germ cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.15.504025v1?rss=1</link>
<description><![CDATA[
Drosophila ovarian germline stem cells (GSCs) are powerful model for stem cell research. However, due to the scarcity of GSCs in ovarian tissue, it is difficult to obtain the transcriptional profile of GSCs and identify novel GSC markers. In this study, we took advantage of single cell RNA sequencing (scRNA-seq) to profile the germline cells and somatic cells in wild type Drosophila ovary. We then performed an in vivo RNAi screen and network analysis to identify genes that are involved in the early stages of germ cell differentiation. We identified 33 genes with limited expression during early germ cell development and identified 19 genes that potentially regulate germ cell differentiation. Among these, an uncharacterized gene, which we named eggplant (eggpl), is highly expressed in GSC and downregulated in early daughter cells. Upon RNAi knockdown of eggpl, we observed an increase in germ cell proliferation, an accumulation of cysts in the early mitotic (2- and 4-cell) stages and an increase in overall ovary size compared to control when flies were maintained on a standard diet. In flies fed a rich yeast diet, the expression of eggpl was significantly lower and the effects of eggpl RNAi were suppressed, suggesting that downregulation of eggpl may link nutritional status to germ cell proliferation and differentiation. We also found that the matrix metalloproteases, Mmp1 and Mmp2 as well as the tissue inhibitor of metalloproteases (Timp) are additional regulators of eggpl. Collectively, this study provides new insight into the signaling networks that regulate early germ cell development and identifies eggpl as a key player in this process.
]]></description>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Nystul, T.</dc:creator>
<dc:creator>Zhong, G.</dc:creator>
<dc:date>2022-08-15</dc:date>
<dc:identifier>doi:10.1101/2022.08.15.504025</dc:identifier>
<dc:title><![CDATA[Single-cell RNA sequencing identifies regulators of differentiation and nutritional cues in Drosophila female germ cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.16.504209v1?rss=1">
<title>
<![CDATA[
Deriving Schwann Cells from hPSCs Enables Disease Modeling and Drug Discovery for Diabetic Peripheral Neuropathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.16.504209v1?rss=1</link>
<description><![CDATA[
Schwann cells (SCs) are the myelinating and non-myelinating glia of the peripheral nervous system (PNS) and are essential for its function. Defects in SCs are associated with many PNS disorders including diabetic peripheral neuropathy (DPN), a condition affecting millions of patients. Here we present a strategy for deriving and purifying SCs from human pluripotent stem cells (hPSCs). The scalable cultures of SCs allow basic and translational applications such as high-resolution molecular and functional characterization, developmental studies, modeling and mechanistic understanding of SC diseases and drug discovery. Our hPSC-derived SCs recapitulate the key molecular features of primary SCs and are capable of engrafting efficiently and producing myelin in injured sciatic nerves in rats. We further established an hPSC-based in vitro model of DPN that revealed the selective vulnerability of human SCs to hyperglycemia-induced cytotoxicity. We established a high-throughput screening system that identified a candidate drug that counteracts glucose-mediated cytotoxicity in SCs and normalizes glucose-induced transcriptional and metabolic abnormalities in SCs. Treatment of hyperglycemic mice with this drug candidate rescues sensory function, prevents SC death, and counteracts myelin damage in sciatic nerves suggesting considerable potential as a novel treatment for DPN.
]]></description>
<dc:creator>Majd, H.</dc:creator>
<dc:creator>Amin, S.</dc:creator>
<dc:creator>Ghazizadeh, Z.</dc:creator>
<dc:creator>Cesiulis, A.</dc:creator>
<dc:creator>Arroyo, E.</dc:creator>
<dc:creator>Lankford, K.</dc:creator>
<dc:creator>Farahvashi, S.</dc:creator>
<dc:creator>Chemel, A. K.</dc:creator>
<dc:creator>Okoye, M.</dc:creator>
<dc:creator>Scantlen, M. D.</dc:creator>
<dc:creator>Tchieu, J.</dc:creator>
<dc:creator>Calder, E. L.</dc:creator>
<dc:creator>LeRouzic, V.</dc:creator>
<dc:creator>Arab, A.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Pasternak, G.</dc:creator>
<dc:creator>Kocsis, J. D.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Studer, L.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:date>2022-08-17</dc:date>
<dc:identifier>doi:10.1101/2022.08.16.504209</dc:identifier>
<dc:title><![CDATA[Deriving Schwann Cells from hPSCs Enables Disease Modeling and Drug Discovery for Diabetic Peripheral Neuropathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.17.504324v1?rss=1">
<title>
<![CDATA[
Arrestin-3 agonism at D3 dopamine receptors defines a subclass of second generation antipsychotics that promotes drug tolerance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.17.504324v1?rss=1</link>
<description><![CDATA[
Second generation antipsychotics (SGAs) are front-line treatments for serious mental illness. Often, individual patients benefit only from some SGAs and not others. The mechanisms underlying this unpredictability in treatment efficacy remain unclear. All SGAs bind the D3 dopamine receptor (D3R) and are traditionally considered antagonists for dopamine receptor signaling. Here, we report that some clinically important SGAs function as arrestin-3 agonists at D3R, resulting in modulation of calcium channels localized to the site of action potential initiation in prefrontal cortex pyramidal neurons. We further show that chronic treatment with an arrestin-3 agonist-SGA, but not an antagonist-SGA, abolishes D3R function through post-endocytic receptor degradation by G-protein coupled receptor-associated sorting protein-1 (GASP1). These results implicate D3R-arrestin-3 signaling as a source of SGA variability, highlighting the importance of including arrestin-3 signaling in characterizations of drug action. Furthermore, they suggest that post-endocytic receptor trafficking that occurs during chronic SGA treatment may contribute to treatment efficacy.
]]></description>
<dc:creator>Schamiloglu, S.</dc:creator>
<dc:creator>Lewis, E.</dc:creator>
<dc:creator>Hergarden, A. C.</dc:creator>
<dc:creator>Bender, K.</dc:creator>
<dc:creator>Whistler, J. L.</dc:creator>
<dc:date>2022-08-18</dc:date>
<dc:identifier>doi:10.1101/2022.08.17.504324</dc:identifier>
<dc:title><![CDATA[Arrestin-3 agonism at D3 dopamine receptors defines a subclass of second generation antipsychotics that promotes drug tolerance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.20.504661v1?rss=1">
<title>
<![CDATA[
Testing the ion-current model for flagellar length sensing and IFT regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.20.504661v1?rss=1</link>
<description><![CDATA[
Eukaryotic cilia and flagella are microtubule-based organelles whose relatively simple shape makes them ideal for investigating the fundamental question of organelle size regulation. Most of the flagellar materials are transported from the cell body via an active transport process called intraflagellar transport (IFT). The rate of IFT entry into flagella, known as IFT injection, has been shown to negatively correlate with flagellar length. However, it remains unknown how the cell measures the length of its flagella and controls IFT injection. One of the most-discussed theoretical models for length sensing to control IFT is the ion-current model, which posits that there is a uniform distribution of Ca2+ channels along the flagellum and that the Ca2+ current from the flagellum into the cell body increases linearly with flagellar length. In this model, the cell uses the Ca2+ current to negatively regulate IFT injection. The recent discovery that IFT entry into flagella is regulated by the phosphorylation of kinesin through a calcium-dependent protein kinase has provided further impetus for the ion-current model. To test this model, we measured and manipulated the levels of Ca2+ inside of Chlamydomonas flagella and quantified IFT injection. Although the concentration of Ca2+ inside of flagella was weakly correlated with the length of flagella, we found that IFT injection was reduced in calcium-deficient flagella, rather than increased as the model predicted, and that variation in IFT injection was uncorrelated with the occurrence of flagellar Ca2+ spikes. Thus, Ca2+ does not appear to function as a negative regulator of IFT injection, hence it cannot form the basis of a stable length control system.
]]></description>
<dc:creator>Ishikawa, H.</dc:creator>
<dc:creator>Moore, J.</dc:creator>
<dc:creator>Diener, D.</dc:creator>
<dc:creator>Delling, M.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2022-08-21</dc:date>
<dc:identifier>doi:10.1101/2022.08.20.504661</dc:identifier>
<dc:title><![CDATA[Testing the ion-current model for flagellar length sensing and IFT regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.22.504863v1?rss=1">
<title>
<![CDATA[
A Mesp1-dependent developmental breakpoint in transcriptional and epigenomic specification of early cardiac precursors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.22.504863v1?rss=1</link>
<description><![CDATA[
Transcriptional networks governing cardiac precursor cell (CPC) specification are incompletely understood due in part to limitations in distinguishing CPCs from non-cardiac mesoderm in early gastrulation. We leveraged detection of early cardiac lineage transgenes within a granular single cell transcriptomic time course of mouse embryos to identify emerging CPCs and describe their transcriptional profiles. Mesp1, a transiently-expressed mesodermal transcription factor (TF), is canonically described as an early regulator of cardiac specification. However, we observed perdurance of CPC transgene-expressing cells in Mesp1 mutants, albeit mis-localized, prompting us to investigate the scope of Mesp1s role in CPC emergence and differentiation. Mesp1 mutant CPCs failed to robustly activate markers of cardiomyocyte maturity and critical cardiac TFs, yet they exhibited transcriptional profiles resembling cardiac mesoderm progressing towards cardiomyocyte fates. Single cell chromatin accessibility analysis defined a Mesp1-dependent developmental breakpoint in cardiac lineage progression at a shift from mesendoderm transcriptional networks to those necessary for cardiac patterning and morphogenesis. These results reveal Mesp1-independent aspects of early CPC specification and underscore a Mesp1-dependent regulatory landscape required for progression through cardiogenesis.
]]></description>
<dc:creator>Krup, A. L.</dc:creator>
<dc:creator>Winchester, S.</dc:creator>
<dc:creator>Ranade, S.</dc:creator>
<dc:creator>Agrawal, A.</dc:creator>
<dc:creator>Devine, W. P.</dc:creator>
<dc:creator>Sinha, T.</dc:creator>
<dc:creator>Choudhary, K.</dc:creator>
<dc:creator>Dominguez, M.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Black, B. L.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Bruneau, B.</dc:creator>
<dc:date>2022-08-22</dc:date>
<dc:identifier>doi:10.1101/2022.08.22.504863</dc:identifier>
<dc:title><![CDATA[A Mesp1-dependent developmental breakpoint in transcriptional and epigenomic specification of early cardiac precursors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.24.505186v1?rss=1">
<title>
<![CDATA[
Defective Ca2+- dependent activation of TRPM4 channels contributes to age-related cerebral small vessel disease in Col4a1 mutant mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.24.505186v1?rss=1</link>
<description><![CDATA[
Humans and mice with mutations in COL4A1 and COL4A2 manifest hallmarks of cerebral small vessel disease (cSVD), but the pathogenic mechanisms are largely unknown. Mice with a missense mutation in Col4a1 at amino acid 1344 (Col4a1+/G1344D) exhibited age-dependent intracerebral hemorrhage (ICH) and brain lesions. Here we report that this pathology was associated with the loss of myogenic vasoconstriction, an intrinsic vascular response essential for the autoregulation of cerebral blood flow. Electrophysiological analyses showed that the loss of myogenic constriction resulted from blunted pressure-induced smooth muscle cell (SMC) membrane depolarization. Further, we found that dysregulation of membrane potential was associated with impaired Ca2+-dependent activation of large-conductance Ca2+-activated K+ (BK) and transient receptor potential melastatin 4 (TRPM4) cation channels linked to disruptions in sarcoplasmic reticulum (SR) Ca2+ signaling. Treating Col4a1+/G1344D mice with 4-phenylbutyrate, a compound that promotes the trafficking of misfolded proteins and alleviates SR stress, restored SR Ca2+ signaling, BK and TRPM4 channel activity, prevented loss of myogenic tone, and reduced ICH. We conclude that alterations in SR Ca2+ handling that impair membrane potential regulating ion channel activity result in dysregulation of SMC membrane potential and loss of myogenic tone contributing to age-related cSVD in Col4a1+/G1344D mice.

Graphical Abstract

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org.highwire.dtl.DTLVardef@11a8cb9org.highwire.dtl.DTLVardef@d00b8org.highwire.dtl.DTLVardef@1aa5b24org.highwire.dtl.DTLVardef@145a9c0_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Yamasaki, E.</dc:creator>
<dc:creator>Thakore, P.</dc:creator>
<dc:creator>Ali, S.</dc:creator>
<dc:creator>Solano, A. S.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Gao, X.</dc:creator>
<dc:creator>Labelle-Dumais, C.</dc:creator>
<dc:creator>Chaumeil, M. M.</dc:creator>
<dc:creator>Gould, D. B.</dc:creator>
<dc:creator>Earley, S.</dc:creator>
<dc:date>2022-08-26</dc:date>
<dc:identifier>doi:10.1101/2022.08.24.505186</dc:identifier>
<dc:title><![CDATA[Defective Ca2+- dependent activation of TRPM4 channels contributes to age-related cerebral small vessel disease in Col4a1 mutant mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.25.505289v1?rss=1">
<title>
<![CDATA[
Organ-Specific Fuel Rewiring in Acute and Chronic Hypoxia Redistributes Glucose and Fatty Acid Metabolism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.25.505289v1?rss=1</link>
<description><![CDATA[
Oxygen deprivation can be detrimental. However, chronic hypoxia is associated with decreased incidence of metabolic syndrome and cardiovascular disease in high-altitude populations. Previously, hypoxic fuel rewiring has primarily been studied in immortalized cells. Here, we describe how systemic hypoxia rewires fuel metabolism to optimize whole-body adaptation. Acclimatization to hypoxia coincided with dramatically lower blood glucose and adiposity. Using in vivo fuel uptake and flux measurements, we found that organs partitioned fuels differently during hypoxia adaption. Acutely, most organs increased glucose uptake and suppressed aerobic glucose oxidation, consistent with previous in vitro investigations. In contrast, brown adipose tissue and skeletal muscle became "glucose savers," suppressing glucose uptake by 3-5-fold. Interestingly, chronic hypoxia produced distinct patterns: the heart relied increasingly on glucose oxidation, and unexpectedly, the brain, kidney, and liver increased fatty acid uptake and oxidation. Hypoxia-induced metabolic plasticity carries therapeutic implications for chronic metabolic diseases and acute hypoxic injuries.
]]></description>
<dc:creator>Midha, A. D.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Queliconi, B. B.</dc:creator>
<dc:creator>Barrios, A. M.</dc:creator>
<dc:creator>Fong, C. O. Y.</dc:creator>
<dc:creator>Blecha, J. E.</dc:creator>
<dc:creator>VanBrocklin, H.</dc:creator>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>Jain, I. H.</dc:creator>
<dc:date>2022-08-26</dc:date>
<dc:identifier>doi:10.1101/2022.08.25.505289</dc:identifier>
<dc:title><![CDATA[Organ-Specific Fuel Rewiring in Acute and Chronic Hypoxia Redistributes Glucose and Fatty Acid Metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.25.505311v1?rss=1">
<title>
<![CDATA[
Genome-wide prediction of disease variants with a deep protein language model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.25.505311v1?rss=1</link>
<description><![CDATA[
Distinguishing between damaging and neutral missense variants is an ongoing challenge in human genetics, with profound implications for clinical diagnosis, genetic studies and protein engineering. Recently, deep-learning models have achieved state-of-the-art performance in classifying variants as pathogenic or benign. However, these models are currently unable to provide predictions over all missense variants, either because of dependency on close protein homologs or due to software limitations. Here we leveraged ESM1b, a 650M-parameter protein language model, to predict the functional impact of human coding variation at scale. To overcome existing technical limitations, we developed a modified ESM1b workflow and functionalized, for the first time, all proteins in the human genome, resulting in predictions for all [~]450M possible missense variant effects. ESM1b was able to distinguish between pathogenic and benign variants across [~]150K variants annotated in ClinVar and HGMD, outperforming existing state-of-the-art methods. ESM1b also exceeded the state of the art at predicting the experimental results of deep mutational scans. We further annotated [~]2M variants across [~]9K alternatively-spliced genes as damaging in certain protein isoforms while neutral in others, demonstrating the importance of considering all isoforms when functionalizing variant effects. The complete catalog of variant effect predictions is available at: https://huggingface.co/spaces/ntranoslab/esm_variants.
]]></description>
<dc:creator>Brandes, N.</dc:creator>
<dc:creator>Goldman, G.</dc:creator>
<dc:creator>Wang, C. H.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Ntranos, V.</dc:creator>
<dc:date>2022-08-26</dc:date>
<dc:identifier>doi:10.1101/2022.08.25.505311</dc:identifier>
<dc:title><![CDATA[Genome-wide prediction of disease variants with a deep protein language model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.25.505318v1?rss=1">
<title>
<![CDATA[
Plasma Proteome Perturbation for CMV DNAemia in Kidney Transplantation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.25.505318v1?rss=1</link>
<description><![CDATA[
BackgroundCytomegalovirus (CMV) infection, either de novo or as reactivation after allotransplantation and chronic immunosuppression, is recognized to cause detrimental alloimmune effects, inclusive of higher susceptibility to graft rejection and substantive impact on chronic graft injury and reduced transplant survival. To obtain further insights into the evolution and pathogenesis of CMV infection in an immunocompromised host we evaluated changes in the circulating host proteome serially, before and after transplantation, and during and after CMV DNA replication (DNAemia), as measured by quantitative polymerase chain reaction (QPCR).

MethodsLC-MS-based proteomics was conducted on 168 serially banked plasma samples, from 62 propensity score-matched kidney transplant recipients. Patients were stratified by CMV replication status into 31 with CMV DNAemia and 31 without CMV DNAemia. Patients had blood samples drawn at protocol times of 3- and 12-months post-transplant. Additionally, blood samples were also drawn before and 1 week and 1 month after detection of CMV DNAemia. Plasma proteins were analyzed using an LCMS 8060 triple quadrupole mass spectrometer. Further, public transcriptomic data on time matched PBMCs samples from the same patients was utilized to evaluate integrative pathways. Data analysis was conducted using R and Limma.

ResultsSamples were segregated based on their proteomic profiles with respect to their CMV Dnaemia status. A subset of 17 plasma proteins was observed to predict the onset of CMV at 3 months post-transplant enriching platelet degranulation (FDR, 4.83E-06), acute inflammatory response (FDR, 0.0018), blood coagulation (FDR, 0.0018) pathways. An increase in many immune complex proteins were observed at CMV infection. Prior to DNAemia the plasma proteome showed changes in the anti-inflammatory adipokine vaspin (SERPINA12), copper binding protein ceruloplasmin (CP), complement activation (FDR=0.03), and proteins enriched in the humoral (FDR=0.01) and innate immune responses (FDR=0.01).

ConclusionPlasma proteomic and transcriptional perturbations impacting humoral and innate immune pathways are observed during CMV infection and provide biomarkers for CMV disease prediction and resolution. Further studies to understand the clinical impact of these pathways can help in the formulation of different types and duration of anti-viral therapies for management of CMV infection in the immunocompromised host.
]]></description>
<dc:creator>Sigdel, T.</dc:creator>
<dc:creator>Boada, P.</dc:creator>
<dc:creator>Kerwin, M.</dc:creator>
<dc:creator>Rashmi, P.</dc:creator>
<dc:creator>Gjertson, D.</dc:creator>
<dc:creator>Rossetti, M.</dc:creator>
<dc:creator>Sur, S.</dc:creator>
<dc:creator>Munar, D.</dc:creator>
<dc:creator>Cimino, J.</dc:creator>
<dc:creator>Ahn, R.</dc:creator>
<dc:creator>Pickering, H.</dc:creator>
<dc:creator>Sen, S.</dc:creator>
<dc:creator>Parmar, R.</dc:creator>
<dc:creator>Fatou, B.</dc:creator>
<dc:creator>Steen, H.</dc:creator>
<dc:creator>Schaenman, J.</dc:creator>
<dc:creator>Bunnapradist, S.</dc:creator>
<dc:creator>Reed, E.</dc:creator>
<dc:creator>Sarwal, M. M.</dc:creator>
<dc:creator>CMV Systems Immunobiology Group,</dc:creator>
<dc:date>2022-08-26</dc:date>
<dc:identifier>doi:10.1101/2022.08.25.505318</dc:identifier>
<dc:title><![CDATA[Plasma Proteome Perturbation for CMV DNAemia in Kidney Transplantation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.27.505518v1?rss=1">
<title>
<![CDATA[
LSD-induced increase of Ising temperature and algorithmic complexity of brain dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.27.505518v1?rss=1</link>
<description><![CDATA[
A topic of growing interest in computational neuroscience is the discovery of fundamental principles underlying global dynamics and the self-organization of the brain. In particular, the notion that the brain operates near criticality has gained considerable support, and recent work has shown that the dynamics of different brain states may be modeled by pairwise maximum entropy Ising models at various distances from a phase transition, i.e., from criticality. Here we aim to characterize two brain states (psychedelics-induced and placebo) as captured by functional magnetic resonance imaging (fMRI), with features derived from the Ising spin model formalism (system temperature, critical point, susceptibility) and from algorithmic complexity. We hypothesized, along the lines of the entropic brain hypothesis, that psychedelics drive brain dynamics into a more disordered state at a higher Ising temperature and increased complexity. We analyze resting state blood-oxygen-level-dependent (BOLD) fMRI data collected in an earlier study from fifteen subjects in a control condition (placebo) and during ingestion of lysergic acid diethylamide (LSD). Working with the automated anatomical labeling (AAL) brain parcellation, we first create "archetype" Ising models representative of the entire dataset (global) and of the data in each condition. Remarkably, we find that such archetypes exhibit a strong correlation with an average structural connectome template obtained from dMRI (r = 0.6). We compare the archetypes from the two conditions and find that the Ising connectivity in the LSD condition is lower than the placebo one, especially at homotopic links (interhemispheric connectivity), reflecting a significant decrease of homotopic functional connectivity in the LSD condition. The global archetype is then personalized for each individual and condition by adjusting the system temperature. The resulting temperatures are all near but above the critical point of the model in the paramagnetic (disordered) phase. The individualized Ising temperatures are higher in the LSD condition than the placebo condition (p = 9 x 10-5). Next, we estimate the Lempel-Ziv-Welch (LZW) complexity of the binarized BOLD data and the synthetic data generated with the individualized model using the Metropolis algorithm for each participant and condition. The LZW complexity computed from experimental data reveals a weak statistical relationship with condition (p = 0.04 one-tailed Wilcoxon test) and none with Ising temperature (r(13) = 0.13, p = 0.65), presumably because of the limited length of the BOLD time series. Similarly, we explore complexity using the block decomposition method (BDM), a more advanced method for estimating algorithmic complexity. The BDM complexity of the experimental data displays a significant correlation with Ising temperature (r(13) = 0.56, p = 0.03) and a weak but significant correlation with condition (p = 0.04, one-tailed Wilcoxon test). This study suggests that the effects of LSD increase the complexity of brain dynamics by loosening interhemispheric connectivity--especially homotopic links. In agreement with earlier work using the Ising formalism with BOLD data, we find the brain state in the placebo condition is already above the critical point, with LSD resulting in a shift further away from criticality into a more disordered state.

Author summaryIn this study, we aim to characterize two brain states (psychedelics-induced and placebo), as captured in functional magnetic resonance imaging (fMRI) data, with features derived from the Ising model formalism (system temperature, critical point, susceptibility) and from algorithmic complexity. Under the hypothesis that psychedelics drive the brain into a more disordered state, we study criticality features of brain dynamics under LSD in a within-subject study using the Ising model formalism and algorithmic complexity using Lempel-Ziv and the Block Decomposition methods. Personalized Ising models are created by first using BOLD fMRI data from all the subjects and conditions to create a single Ising "archetype" model--which we can interpret as the average model of the data at unit temperature--and then by adjusting the model temperature for each subject and condition. We find that the effects of LSD translate into increased BOLD signal complexity and Ising temperature, in agreement with earlier findings and predictions from existing theories of the effects of psychedelics, such as the relaxed beliefs under psychedelics (REBUS), the anarchic brain hypothesis [1], and the algorithmic information theory of consciousness (KT) [2, 3]. However, in contrast with some of the previously cited theories, we find that the system in the placebo condition is already in the paramagnetic phase--above the critical point--with ingestion of LSD resulting in a shift away from Ising criticality into a more disordered state. Finally, we highlight the fact that the structural connectome can be recovered to a good degree by fitting an Ising model and that the reduction of homotopic links appears to play an important role in the slide to disorder under psychedelics.
]]></description>
<dc:creator>Ruffini, G.</dc:creator>
<dc:creator>Damiani, G.</dc:creator>
<dc:creator>Lozano-Soldevilla, D.</dc:creator>
<dc:creator>Deco, N.</dc:creator>
<dc:creator>Rosas, F.</dc:creator>
<dc:creator>Kiani, N.</dc:creator>
<dc:creator>Ponce-Alvarez, A.</dc:creator>
<dc:creator>Kringelbach, M. L.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Deco, G. L.</dc:creator>
<dc:date>2022-08-29</dc:date>
<dc:identifier>doi:10.1101/2022.08.27.505518</dc:identifier>
<dc:title><![CDATA[LSD-induced increase of Ising temperature and algorithmic complexity of brain dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/203224v1?rss=1">
<title>
<![CDATA[
Genome-wide and high-density CRISPR-Cas9 screens identify point mutations in PARP1 causing PARP inhibitor resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/203224v1?rss=1</link>
<description><![CDATA[
PARP inhibitors (PARPi) target homologous recombination defective tumour cells via synthetic lethality. Genome-wide and high-density CRISPR-Cas9 "tag, mutate and enrich" mutagenesis screens identified single amino acid mutations in PARP1 that cause profound PARPi-resistance. These included PARP1 mutations outside of the DNA interacting regions of the protein, such as mutations in solvent exposed regions of the catalytic domain and clusters of mutations around points of contact between ZnF, WGR and HD domains. These mutations altered PARP1 trapping, as did a mutation found in a clinical case of PARPi resistance. These genetic studies reinforce the importance of trapped PARP1 as a key cytotoxic DNA lesion and suggest that interactions between non-DNA binding domains of PARP1 influence cytotoxicity. Finally, different mechanisms of PARPi resistance (BRCA1 reversion, PARP1, 53BP1, REV7 mutation) had differing effects on chemotherapy sensitivity, suggesting that the underlying mechanism of PARPi resistance likely influences the success of subsequent therapies.
]]></description>
<dc:creator>Pettitt, S. J.</dc:creator>
<dc:creator>Krastev, D. B.</dc:creator>
<dc:creator>Brandsma, I.</dc:creator>
<dc:creator>Drean, A.</dc:creator>
<dc:creator>Song, F.</dc:creator>
<dc:creator>Aleksandrov, R.</dc:creator>
<dc:creator>Harrell, M. I.</dc:creator>
<dc:creator>Menon, M.</dc:creator>
<dc:creator>Brough, R.</dc:creator>
<dc:creator>Campbell, J.</dc:creator>
<dc:creator>Frankum, J.</dc:creator>
<dc:creator>Ranes, M.</dc:creator>
<dc:creator>Pemberton, H. N.</dc:creator>
<dc:creator>Rafiq, R.</dc:creator>
<dc:creator>Fenwick, K.</dc:creator>
<dc:creator>Swain, A.</dc:creator>
<dc:creator>Guettler, S.</dc:creator>
<dc:creator>Lee, J.-M.</dc:creator>
<dc:creator>Swisher, E. M.</dc:creator>
<dc:creator>Stoynov, S.</dc:creator>
<dc:creator>Yusa, K.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Lord, C. J.</dc:creator>
<dc:date>2017-10-14</dc:date>
<dc:identifier>doi:10.1101/203224</dc:identifier>
<dc:title><![CDATA[Genome-wide and high-density CRISPR-Cas9 screens identify point mutations in PARP1 causing PARP inhibitor resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/204693v1?rss=1">
<title>
<![CDATA[
Light-Activated Cell Identification and Sorting (LACIS): A New Method to Identify and Select Edited Clones on a Microfluidic Device 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/204693v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas9 gene editing has revolutionized cell engineering and promises to open new doors in gene and cell therapies. Despite improvements in the CRISPR-editing molecular toolbox in cell lines and primary cells, identifying and purifying properly edited clones remains slow, laborious and low-yield. Here, we establish a new method that uses cell manipulation on a chip with Opto-Electronic Positioning (OEP) technology to enable clonal isolation and selection of edited cells. We focused on editing CXCR4 in primary human T cells, a gene that encodes a co-receptor for HIV entry. T cells hold significant potential for cell-based therapy, but the gene-editing efficiency and expansion potential of these cells is limited. We describe here a method to obviate these limitations. Briefly, after electroporation of cells with CXCR4-targeting Cas9 ribonucleoproteins (RNPs), single T cells were isolated on a chip, where they proliferated over time into well-resolved colonies. Phenotypic consequences of genome editing could be rapidly assessed on-chip with cell-surface staining for CXCR4. Furthermore, independent of phenotype, individual colonies could be identified based on their specific genotype at the 5-10 cell stage. Each colony was split and sequentially exported for immediate on-target sequencing and validation, and further off-chip clonal expansion of the validated clones. We were able to assess single-clone editing efficiencies, including the rate of monoallelic and biallelic indels or precise nucleotide replacements. This new method will enable identification and selection of perfectly edited clones within 10 days from Cas9-RNP introduction in cells based on the phenotype and/or genotype.
]]></description>
<dc:creator>Mocciaro, A.</dc:creator>
<dc:creator>Roth, T.</dc:creator>
<dc:creator>Bennett, H.</dc:creator>
<dc:creator>Soumillon, M.</dc:creator>
<dc:creator>Shah, A.</dc:creator>
<dc:creator>Hiatt, J.</dc:creator>
<dc:creator>Chapman, K.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Lavieu, G.</dc:creator>
<dc:date>2017-10-17</dc:date>
<dc:identifier>doi:10.1101/204693</dc:identifier>
<dc:title><![CDATA[Light-Activated Cell Identification and Sorting (LACIS): A New Method to Identify and Select Edited Clones on a Microfluidic Device]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/208462v1?rss=1">
<title>
<![CDATA[
Hsp90 dictates viral sequence space by balancing the evolutionary tradeoffs between protein stability, aggregation and translation rate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/208462v1?rss=1</link>
<description><![CDATA[
Acquisition of mutations is central to evolution but the detrimental effects of most mutations on protein folding and stability limit protein evolvability. Molecular chaperones, which suppress aggregation and facilitate polypeptide folding, are proposed to promote sequence diversification by buffering destabilizing mutations. However, whether and how chaperones directly control protein evolution remains poorly understood. Here, we examine the effect of reducing the activity of the key eukaryotic chaperone Hsp90 on poliovirus evolution. Contrary to predictions of a buffering model, inhibiting Hsp90 increases population sequence diversity and promotes accumulation of mutations reducing protein stability. Explaining this counterintuitive observation, we find that Hsp90 offsets the evolutionary tradeoff between protein stability and aggregation. Lower chaperone levels favor sequence variants of reduced hydrophobicity, thus decreasing protein aggregation propensity but at a cost to protein stability. Notably, reducing Hsp90 activity also promotes clusters of codon-deoptimized synonymous mutations at inter-domain boundaries, likely to promote local ribosomal slowdown to facilitate cotranslational domain folding. Our results reveal how a chaperone can shape the sequence landscape at both the protein and RNA levels to harmonize the competing constraints posed by protein stability, aggregation propensity and translation rate on successful protein biogenesis.
]]></description>
<dc:creator>Geller, R.</dc:creator>
<dc:creator>Pechmann, S.</dc:creator>
<dc:creator>Acevedo, A.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:date>2017-10-24</dc:date>
<dc:identifier>doi:10.1101/208462</dc:identifier>
<dc:title><![CDATA[Hsp90 dictates viral sequence space by balancing the evolutionary tradeoffs between protein stability, aggregation and translation rate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/209478v1?rss=1">
<title>
<![CDATA[
Robust identification of deletions in exome and genome sequence data based on clustering of Mendelian errors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/209478v1?rss=1</link>
<description><![CDATA[
Multiple tools have been developed to identify copy number variants (CNVs) from whole exome (WES) and whole genome sequencing (WGS) data. Current tools such as XHMM for WES and CNVnator for WGS identify CNVs based on changes in read depth. For WGS, other methods to identify CNVs include utilizing discordant read pairs and split reads and genome-wide local assembly with tools such as Lumpy and SvABA, respectively. Here, we introduce a new method to identify deletion CNVs from WES and WGS trio data based on the clustering of Mendelian errors (MEs). Using our Mendelian Error Method (MEM), we identified 127 deletions (inherited and de novo) in 2,601 WES trios from the Pediatric Cardiac Genomics Consortium, with a validation rate of 88% by digital droplet PCR. MEM identified additional de novo deletions compared to XHMM, and also identified sample switches, DNA contamination, a significant enrichment of 15q11.2 deletions compared to controls and eight cases of uniparental disomy. We applied MEM to WGS data from the Genome In A Bottle Ashkenazi trio and identified deletions with 97% specificity. MEM provides a robust, computationally inexpensive method for identifying deletions, and an orthogonal approach for verifying deletions called by other tools.
]]></description>
<dc:creator>Manheimer, K. B.</dc:creator>
<dc:creator>Patel, N.</dc:creator>
<dc:creator>Richter, F.</dc:creator>
<dc:creator>Gorham, J.</dc:creator>
<dc:creator>Tai, A. C.</dc:creator>
<dc:creator>Homsy, J.</dc:creator>
<dc:creator>Boskovski, M. T.</dc:creator>
<dc:creator>Parfenov, M.</dc:creator>
<dc:creator>Goldmuntz, E.</dc:creator>
<dc:creator>Chung, W. K.</dc:creator>
<dc:creator>Brueckner, M.</dc:creator>
<dc:creator>Tristani-Firouzi, M.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Seidman, J. G.</dc:creator>
<dc:creator>Seidman, C. E.</dc:creator>
<dc:creator>Gelb, B. D.</dc:creator>
<dc:creator>Sharp, A. J.</dc:creator>
<dc:date>2017-10-26</dc:date>
<dc:identifier>doi:10.1101/209478</dc:identifier>
<dc:title><![CDATA[Robust identification of deletions in exome and genome sequence data based on clustering of Mendelian errors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/211714v1?rss=1">
<title>
<![CDATA[
A size-invariant bud-length timer enables robustness in yeast cell size control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/211714v1?rss=1</link>
<description><![CDATA[
Cell populations across nearly all forms of life generally maintain a characteristic cell type-dependent size, but how size control is achieved has been a long-standing question. Prior work has uncovered diverse size control strategies operating at distinct cell cycle stages, but it is unclear how these numerous pathways are integrated to provide robust, systems-level cell size control for any organism. Here, we probe cell growth and size control in budding yeast that can be reversibly blocked from bud initiation. While blocked, cells continue to grow isotropically, increasing their volume by more than an order of magnitude over unperturbed cells. Upon release, these  giant yeast resume budding and the population returns to its initial volume distribution within a few cell division cycles. Size control under these conditions does not require an explicit molecular size sensor. Instead, our observations are consistent with a size-invariant bud growth timer specifying the duration of S/G2/M to limit daughter cell size.
]]></description>
<dc:creator>Allard, C. A. H.</dc:creator>
<dc:creator>Decker, F.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:creator>Toettcher, J. E.</dc:creator>
<dc:creator>Graziano, B. R.</dc:creator>
<dc:date>2017-10-31</dc:date>
<dc:identifier>doi:10.1101/211714</dc:identifier>
<dc:title><![CDATA[A size-invariant bud-length timer enables robustness in yeast cell size control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/213553v1?rss=1">
<title>
<![CDATA[
A multi-center study on factors influencing the reproducibility of in vitro drug-response studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/213553v1?rss=1</link>
<description><![CDATA[
Evidence that some influential biomedical results cannot be repeated has increased interest in practices that generate data meeting findable, accessible, interoperable and reproducible (FAIR) standards. Multiple papers have identified examples of irreproducibility, but practical steps for increasing reproducibility have not been widely studied. Here, seven research centers in the NIH LINCS Program Consortium investigate the reproducibility of a prototypical perturbational assay: quantifying the responsiveness of cultured cells to anti-cancer drugs. Such assays are important for drug development, studying cell biology, and patient stratification. While many experimental and computational factors have an impact on intra- and inter-center reproducibility, the factors most difficult to identify and correct are those with a strong dependency on biological context. These factors often vary in magnitude with the drug being analyzed and with growth conditions. We provide ways of identifying such context-sensitive factors, thereby advancing the conceptual and practical basis for greater experimental reproducibility.
]]></description>
<dc:creator>Niepel, M.</dc:creator>
<dc:creator>Hafner, M.</dc:creator>
<dc:creator>Williams, E. H.</dc:creator>
<dc:creator>Chung, M.</dc:creator>
<dc:creator>Barrette, A. M.</dc:creator>
<dc:creator>Stern, A. D.</dc:creator>
<dc:creator>Hu, B.</dc:creator>
<dc:creator>LINCS Consortium,</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Birtwistle, M. R.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:date>2017-11-03</dc:date>
<dc:identifier>doi:10.1101/213553</dc:identifier>
<dc:title><![CDATA[A multi-center study on factors influencing the reproducibility of in vitro drug-response studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/213561v1?rss=1">
<title>
<![CDATA[
Metagenomics for chronic meningitis: clarifying interpretation and diagnosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/213561v1?rss=1</link>
<description><![CDATA[
ImportanceIdentifying infectious causes of subacute and chronic meningitis can be challenging. Enhanced, unbiased diagnostic approaches are needed.nnObjectiveTo present a case series of patients with diagnostically challenging subacute and chronic meningitis in whom metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid (CSF), supported by a statistical framework generated from mNGS sequencing of non-infectious patients and environmental controls, identified a pathogen.nnDesignCase series. Using mNGS data from the CSF of 94 non-infectious neuroinflammatory cases and 24 water and reagent controls, we developed and implemented a weighted scoring metric based on z-scores at the species and genus level for both nucleotide and protein databases to prioritize and rank mNGS results. We performed mNGS on total RNA extracted from CSF of patients with subacute or chronic meningitis and highlight seven cases representing a diverse array of pathogens.nnSettingA multi-center study of mNGS pathogen discovery in patients with suspected neuroinflammatory conditions.nnParticipantsPatients with diagnostically challenging subacute or chronic meningitis enrolled in a research study of mNGS performed on CSF.nnInterventionmNGS was performed on total RNA extracted from CSF (0.25-0.5 mL). A weighted z-score was used to filter out environmental contaminants and facilitate efficient data triage and analysis.nnMain Outcomes1) Pathogens identified by mNGS and 2) ability of a statistical model to prioritize, rank, and simplify mNGS results.nnResultsmNGS identified parasitic worms, fungi and viruses in seven subjects: Taenia solium (n=2), Cryptococcus neoformans, human immunodeficiency virus-1, Aspergillus oryzae, Histoplasma capsulatum, and Candida dubliniensis. Evaluating mNGS data with a weighted z-score based scoring algorithm effectively separated bona fide pathogen sequences from spurious environmental sequences.nnConclusions and RelevancemNGS of CSF identified a diversity of microbial pathogens in patients with diagnostically challenging subacute or chronic meningitis, including a case of subarachnoid neurocysticercosis that defied diagnosis for one year, the first case of CNS vasculitis caused by Aspergillus oryzae, and the fourth reported case of Candida dubliniensis meningitis. Filtering metagenomic data with a scoring algorithm greatly clarified data interpretation and highlights the difficulties attributing biological significance to organisms that may be present in control samples used for metagenomic sequencing studies.nnKey PointsQuestion: How can metagenomic next-generation sequencing of cerebrospinal fluid be leveraged to aid in the diagnosis of patients with subacute or chronic meningitis?nnFindings: Metagenomic next-generation sequencing identified parasitic worms, fungi and viruses in a case series of seven subjects. A database of water-only and healthy patient controls enabled application of a z-score based scoring algorithm to effectively separate bona fide pathogen sequences from spurious environmental sequences.nnMeaning: Our scoring algorithm greatly simplified data interpretation in a series of patients with a wide range of challenging infectious causes of subacute or chronic meningitis identified by metagenomic next-generation sequencing.
]]></description>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:creator>O'Donovan, B. D.</dc:creator>
<dc:creator>Gelfand, J. M.</dc:creator>
<dc:creator>Sample, H. A.</dc:creator>
<dc:creator>Chow, F. C.</dc:creator>
<dc:creator>Betjemann, J. P.</dc:creator>
<dc:creator>Shah, M. P.</dc:creator>
<dc:creator>Richie, M. B.</dc:creator>
<dc:creator>Gorman, M. P.</dc:creator>
<dc:creator>Hajj-Ali, R. A.</dc:creator>
<dc:creator>Calabrese, L. H.</dc:creator>
<dc:creator>Zorn, K. C.</dc:creator>
<dc:creator>Greenlee, J. E.</dc:creator>
<dc:creator>Blum, J. H.</dc:creator>
<dc:creator>Green, G.</dc:creator>
<dc:creator>Khan, L. M.</dc:creator>
<dc:creator>Banerji, D.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>Bryson-Cahn, C.</dc:creator>
<dc:creator>Harrington, W.</dc:creator>
<dc:creator>Lingappa, J. R.</dc:creator>
<dc:creator>Shanbhag, N. M.</dc:creator>
<dc:creator>Green, A. J.</dc:creator>
<dc:creator>Brew, B. J.</dc:creator>
<dc:creator>Soldatos, A.</dc:creator>
<dc:creator>Strnad, L.</dc:creator>
<dc:creator>Doernberg, S. B.</dc:creator>
<dc:creator>Jay, C. A.</dc:creator>
<dc:creator>Douglas, V.</dc:creator>
<dc:creator>Josephson, S. A.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:date>2017-11-07</dc:date>
<dc:identifier>doi:10.1101/213561</dc:identifier>
<dc:title><![CDATA[Metagenomics for chronic meningitis: clarifying interpretation and diagnosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/215202v1?rss=1">
<title>
<![CDATA[
Protection from epidemics is a driving force for evolution of lifespan setpoints 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/215202v1?rss=1</link>
<description><![CDATA[
Most living organisms age, as determined by species-specific limits to lifespan1-6. The biological driving force for a genetically-defined limit on the lifespan of a given species (herein called "lifespan setpoint") remains poorly understood. Here we present mathematical models suggesting that an upper limit of individual lifespans protects their cohort population from infection-associated penalties. A shorter lifespan setpoint helps control pathogen spread within a population, prevents the establishment and progression of infections, and accelerates pathogen clearance from the population when compared to populations with long-lived individuals. Strikingly, shorter-living variants efficiently displace longer-living individuals in populations that are exposed to pathogens and exist in spatially structured niches. The beneficial effects of shorter lifespan setpoints are even more evident in the context of zoonotic transmissions, where pathogens undergo adaptation to a new host. We submit that the selective pressure of infectious disease provides an evolutionary driving force to limit individual lifespan setpoints after reproductive maturity to secure its kins fitness. Our findings have important public health implications for efforts to extend humans lifespan.
]]></description>
<dc:creator>Lidsky, P. V.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:date>2017-11-06</dc:date>
<dc:identifier>doi:10.1101/215202</dc:identifier>
<dc:title><![CDATA[Protection from epidemics is a driving force for evolution of lifespan setpoints]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/217968v1?rss=1">
<title>
<![CDATA[
Polymerase pausing induced by sequence-specific RNA binding protein drives heterochromatin assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/217968v1?rss=1</link>
<description><![CDATA[
Packaging of pericentromeric DNA into heterochromatin is crucial for genome stability, development and health, yet its endogenous triggers remain poorly understood1. A defining feature of pericentromeric heterochromatin is histone H3 lysine 9 methylation (H3K9me)2-4. In S. pombe, transcripts derived from the pericentromeric dg and dh repeat during S phase5-7 promote heterochromatin formation through two pathways: an RNAi-dependent mechanism involving recruitment of the Clr4 H3K9 methyltransferase complex (CLR-C) via the RITS complex8-13, and RNAi-independent mechanism involving an RNAPII-associated RNA-binding protein Seb1, the repressor complex SHREC, and RNA processing activities14-19. We show here that Seb1 promotes long-lived RNAPII pausing. Pause sites associated with sequence-specific Seb1 RNA binding events are significantly enriched in pericentromeric repeat regions and their presence correlates with the heterochromatin-triggering activities of the corresponding dg and dh DNA fragments. Remarkably, globally increasing RNAPII stalling by other means induces the formation of novel large ectopic heterochromatin domains. Such ectopic heterochromatin occurs even in cells lacking functional RITS, demonstrating that RNAPII pausing can be sufficient to trigger de novo heterochromatin independently of RNAi. These results uncover Seb1-mediated polymerase stalling as a new signal for nucleating heterochromatin assembly in repetitive DNA.
]]></description>
<dc:creator>Parsa, J.-Y.</dc:creator>
<dc:creator>Boudoukha, S.</dc:creator>
<dc:creator>Burke, J.</dc:creator>
<dc:creator>Homer, C.</dc:creator>
<dc:creator>Madhani, H.</dc:creator>
<dc:date>2017-11-11</dc:date>
<dc:identifier>doi:10.1101/217968</dc:identifier>
<dc:title><![CDATA[Polymerase pausing induced by sequence-specific RNA binding protein drives heterochromatin assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/218966v1?rss=1">
<title>
<![CDATA[
New routes for PTP1B allosteric inhibition by multitemperature crystallography, fragment screening and covalent tethering 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/218966v1?rss=1</link>
<description><![CDATA[
Allostery is an inherent feature of proteins and provides alternative routes to regulating function. Small-molecule allosteric inhibitors are often desirable; however, it remains challenging to identify surface sites in proteins which can bind small molecules and modulate function. We identified new allosteric sites in protein tyrosine phosphatase 1B (PTP1B) by combining multiple-temperature X-ray crystallography experiments and structure determination from hundreds of individual small-molecule fragment soaks. New modeling approaches reveal "hidden" low-occupancy conformational states for protein and ligands. Our results converge on a new allosteric site that is conformationally coupled to the active-site WPD loop, a hotspot for fragment binding, not conserved in the closest homolog, and distinct from other recently reported allosteric sites in PTP1B. Targeting this site with covalently tethered molecules allosterically inhibits enzyme activity. Overall, this work demonstrates how the ensemble nature of macromolecular structure revealed by multitemperature crystallography can be exploited for developing allosteric modulators.
]]></description>
<dc:creator>Keedy, D. A.</dc:creator>
<dc:creator>Hill, Z. B.</dc:creator>
<dc:creator>Biel, J. T.</dc:creator>
<dc:creator>Kang, E.</dc:creator>
<dc:creator>Rettenmaier, T. J.</dc:creator>
<dc:creator>Brandao-Neto, J.</dc:creator>
<dc:creator>Pearce, N. M.</dc:creator>
<dc:creator>von Delft, F.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2017-11-13</dc:date>
<dc:identifier>doi:10.1101/218966</dc:identifier>
<dc:title><![CDATA[New routes for PTP1B allosteric inhibition by multitemperature crystallography, fragment screening and covalent tethering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/219113v1?rss=1">
<title>
<![CDATA[
Bringing Diffuse X-ray Scattering Into Focus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/219113v1?rss=1</link>
<description><![CDATA[
X-ray crystallography is experiencing a renaissance as a method for probing the protein conformational ensemble. The inherent limitations of Bragg analysis, however, which only reveals the mean structure, have given way to a surge in interest in diffuse scattering, which is caused by structure variations. Diffuse scattering is present in all macromolecular crystallography experiments. Recent studies are shedding light on the origins of diffuse scattering in protein crystallography, and provide clues for leveraging diffuse scattering to model protein motions with atomic detail.
]]></description>
<dc:creator>Wall, M.</dc:creator>
<dc:creator>Wolff, A. M.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2017-11-13</dc:date>
<dc:identifier>doi:10.1101/219113</dc:identifier>
<dc:title><![CDATA[Bringing Diffuse X-ray Scattering Into Focus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/219238v1?rss=1">
<title>
<![CDATA[
Singleton Variants Dominate the Genetic Architecture of Human Gene Expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/219238v1?rss=1</link>
<description><![CDATA[
The vast majority of human mutations have minor allele frequencies (MAF) under 1%, with the plurality observed only once (i.e., "singletons"). While Mendelian diseases are predominantly caused by rare alleles, their cumulative contribution to complex phenotypes remains largely unknown. We develop and rigorously validate an approach to jointly estimate the contribution of all alleles, including singletons, to phenotypic variation. We apply our approach to transcriptional regulation, an intermediate between genetic variation and complex disease. Using whole genome DNA and lymphoblastoid cell line RNA sequencing data from 360 European individuals, we conservatively estimate that singletons contribute ~25% of cis-heritability across genes (dwarfing the contributions of other frequencies). Strikingly, the majority (~76%) of singleton heritability derives from ultra-rare variants absent from thousands of additional samples. We develop a novel inference procedure to demonstrate that our results are consistent with rampant purifying selection shaping the regulatory architecture of most human genes.
]]></description>
<dc:creator>Hernandez, R. D.</dc:creator>
<dc:creator>Uricchio, L. H.</dc:creator>
<dc:creator>Hartman, K.</dc:creator>
<dc:creator>Ye, J.</dc:creator>
<dc:creator>Dahl, A.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:date>2017-11-14</dc:date>
<dc:identifier>doi:10.1101/219238</dc:identifier>
<dc:title><![CDATA[Singleton Variants Dominate the Genetic Architecture of Human Gene Expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/222901v1?rss=1">
<title>
<![CDATA[
Cytoplasmic import and processing of mRNA amplify transcriptional bursts accounting for the majority of cellular noise 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/222901v1?rss=1</link>
<description><![CDATA[
Transcription is an episodic process characterized by probabilistic bursts; but how these bursts are modulated by cellular physiology remains unclear and has implications for how selection may act on these fluctuations. Using simulations and single-molecule RNA counting, we examined how cellular processes influence cell-to-cell variability (noise). The results show that RNA noise is amplified in the cytoplasm compared to the nucleus in ~85% of genes across diverse promoters, genomic loci, and cell types (human and mouse). Surprisingly, measurements show further amplification of RNA noise in the cytoplasm, fitting a model of biphasic mRNA conversion between translation- and degradation-competent states. The multi-state translation-degradation of mRNA also causes substantial noise amplification in protein levels, ultimately accounting for ~74% of intrinsic protein variability in cell populations. Overall, the results demonstrate how transcriptional bursts are intrinsically amplified by mRNA processing and indicate mechanisms through which noise could act as a substrate for evolutionary selection.
]]></description>
<dc:creator>Hansen, M. M. K.</dc:creator>
<dc:creator>Desai, R. V.</dc:creator>
<dc:creator>Simpson, M. L.</dc:creator>
<dc:creator>Weinberger, L. S.</dc:creator>
<dc:date>2017-11-21</dc:date>
<dc:identifier>doi:10.1101/222901</dc:identifier>
<dc:title><![CDATA[Cytoplasmic import and processing of mRNA amplify transcriptional bursts accounting for the majority of cellular noise]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/224170v1?rss=1">
<title>
<![CDATA[
Spliceosome profiling visualizes the operations of a dynamic RNP in vivo at nucleotide resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/224170v1?rss=1</link>
<description><![CDATA[
Tools to understand how the spliceosome functions in vivo have lagged behind advances in its structural biology. We describe methods to globally profile spliceosome-bound precursor, intermediates and products at nucleotide resolution. We apply these tools to three divergent yeast species that span 600 million years of evolution. The sensitivity of the approach enables detection of novel cases of non-canonical catalysis including interrupted, recursive and nested splicing. Employing statistical modeling to understand the quantitative relationships between RNA features and the data, we uncover independent roles for intron size, position and number in substrate progression through the two catalytic stages. These include species-specific inputs suggestive of spliceosome-transcriptome coevolution. Further investigations reveal ATP-dependent discard of numerous endogenous substrates at both the precursor and lariat-intermediate stages and connect discard to intron retention, a form of splicing regulation. Spliceosome profiling is a quantitative, generalizable global technology to investigate an RNP central to eukaryotic gene expression.nnHighlightsO_LIMeasurement of spliceosome-bound precursor and intermediate in three speciesnC_LIO_LINon-canonical splicing events revealednC_LIO_LIStatistical modeling uncovers substrate features that predict catalytic efficiencynC_LIO_LIDiscard of suboptimal substrates occurs in vivo and predicts intron-retained mRNAsnC_LI
]]></description>
<dc:creator>Burke, J.</dc:creator>
<dc:creator>Longhurst, A.</dc:creator>
<dc:creator>Merkurjev, D.</dc:creator>
<dc:creator>Sales-Lee, J.</dc:creator>
<dc:creator>Rao, B.</dc:creator>
<dc:creator>Moresco, J.</dc:creator>
<dc:creator>Yates, J.</dc:creator>
<dc:creator>Li, J. J.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:date>2017-11-22</dc:date>
<dc:identifier>doi:10.1101/224170</dc:identifier>
<dc:title><![CDATA[Spliceosome profiling visualizes the operations of a dynamic RNP in vivo at nucleotide resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/224345v1?rss=1">
<title>
<![CDATA[
Tracing cellular heterogeneity in pooled genetic screens via multi-level barcoding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/224345v1?rss=1</link>
<description><![CDATA[
While pooled loss- and gain-of-function screening approaches have become increasingly popular to systematically investigate mammalian gene function, they have thus far ignored the fact that cell populations are heterogeneous. Here we introduce multi-level barcoded sgRNA libraries to (i) monitor differences in the behavior of multiplexed clonal cell lines, (ii) trace sub-clonal lineages of cells expressing the same sgRNA, (iii) derive in-sample screen replicates and (iv) reduce the number of cells and sequencing read counts required to reach statistical significance. Using our approach, we illustrate how clonal heterogeneity impairs the results of pooled genetic screens and demonstrate the ability of multi-level barcoding to resolve cellular heterogeneity related issues.
]]></description>
<dc:creator>Boettcher, M.</dc:creator>
<dc:creator>Covarrubias, S.</dc:creator>
<dc:creator>Biton, A.</dc:creator>
<dc:creator>Blau, J. A.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:creator>McManus, M. T.</dc:creator>
<dc:date>2017-11-26</dc:date>
<dc:identifier>doi:10.1101/224345</dc:identifier>
<dc:title><![CDATA[Tracing cellular heterogeneity in pooled genetic screens via multi-level barcoding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/226019v1?rss=1">
<title>
<![CDATA[
Developmental loci harbor clusters of accelerated regions that evolved independently in ape lineages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/226019v1?rss=1</link>
<description><![CDATA[
Some of the fastest evolving regions of the human genome are conserved non-coding elements with many human-specific DNA substitutions. These Human Accelerated Regions (HARs) are enriched nearby regulatory genes, and several HARs function as developmental enhancers. To investigate if this evolutionary signature is unique to humans, we quantified evidence of accelerated substitutions in conserved genomic elements across multiple lineages and applied this approach simultaneously to the genomes of five apes: human, chimpanzee, gorilla, orangutan, and gibbon. We find roughly similar numbers and genomic distributions of lineage-specific accelerated regions (linARs) in all five apes. In particular, apes share an enrichment of linARs in regulatory DNA nearby genes involved in development, especially transcription factors and other regulators. Many developmental loci harbor clusters of nonoverlapping linARs from multiple apes, suggesting that accelerated evolution in each species affected distinct regulatory elements that control a shared set of developmental pathways. Our statistical tests distinguish between GC-biased and unbiased accelerated substitution rates, allowing us to quantify the roles of different evolutionary forces in creating linARs. We find evidence of GC-biased gene conversion in each ape, but unbiased acceleration consistent with positive selection or loss of constraint is more common in all five lineages. It therefore appears that similar evolutionary processes created independent accelerated regions in the genomes of different apes, and that these lineage-specific changes to conserved non-coding sequences may have differentially altered expression of a core set of developmental genes across ape evolution.
]]></description>
<dc:creator>Kostka, D.</dc:creator>
<dc:creator>Holloway, A. K.</dc:creator>
<dc:creator>Pollard, K.</dc:creator>
<dc:date>2017-11-28</dc:date>
<dc:identifier>doi:10.1101/226019</dc:identifier>
<dc:title><![CDATA[Developmental loci harbor clusters of accelerated regions that evolved independently in ape lineages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/234740v1?rss=1">
<title>
<![CDATA[
High accuracy measurements of nanometer-scale distances between fluorophores at the single-molecule level 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/234740v1?rss=1</link>
<description><![CDATA[
To uncover the mechanisms of molecular machines it is useful to probe their structural conformations. Single-molecule Forster resonance energy transfer (smFRET) is a powerful tool for measuring intra-molecular shape changes of single-molecules, but is confined to distances of 2-8 nm. Current super-resolution measurements are error prone at <25 nm. Thus, reliable high-throughput distance information between 8-25 nm is currently difficult to achieve. Here, we describe methods that utilize information about localization and imaging errors to measure distances between two different color fluorophores with [~]1 nm accuracy at any distance >2 nm, using a standard TIRF microscope and open-source software. We applied our two-color localization method to uncover a [~]4 nm conformational change in the "stalk" of the motor protein dynein, revealing unexpected flexibility in this antiparallel coiled-coil domain. These new methods enable high-accuracy distance measurements of single-molecules that can be used over a wide range of length scales.
]]></description>
<dc:creator>Niekamp, S.</dc:creator>
<dc:creator>Sung, J.</dc:creator>
<dc:creator>Huynh, W.</dc:creator>
<dc:creator>Vale, R. D.</dc:creator>
<dc:creator>Stuurman, N.</dc:creator>
<dc:date>2017-12-15</dc:date>
<dc:identifier>doi:10.1101/234740</dc:identifier>
<dc:title><![CDATA[High accuracy measurements of nanometer-scale distances between fluorophores at the single-molecule level]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/235234v1?rss=1">
<title>
<![CDATA[
A Comprehensive Evaluation of the Genetic Architecture of Sudden Cardiac Arrest 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/235234v1?rss=1</link>
<description><![CDATA[
BackgroundSudden cardiac arrest (SCA) accounts for 10% of adult mortality in Western populations. While several risk factors are observationally associated with SCA, the genetic architecture of SCA in the general population remains unknown. Furthermore, understanding which risk factors are causal may help target prevention strategies.nnMethodsWe carried out a large genome-wide association study (GWAS) for SCA (n=3,939 cases, 25,989 non-cases) to examine common variation genome-wide and in candidate arrhythmia genes. We also exploited Mendelian randomization methods using cross-trait multi-variant genetic risk score associations (GRSA) to assess causal relationships of 18 risk factors with SCA.nnResultsNo variants were associated with SCA at genome-wide significance, nor were common variants in candidate arrhythmia genes associated with SCA at nominal significance. Using cross-trait GRSA, we established genetic correlation between SCA and (1) coronary artery disease (CAD) and traditional CAD risk factors (blood pressure, lipids, and diabetes), (2) height and BMI, and (3) electrical instability traits (QT and atrial fibrillation), suggesting etiologic roles for these traits in SCA risk.nnConclusionsOur findings show that a comprehensive approach to the genetic architecture of SCA can shed light on the determinants of a complex life-threatening condition with multiple influencing factors in the general population. The results of this genetic analysis, both positive and negative findings, have implications for evaluating the genetic architecture of patients with a family history of SCA, and for efforts to prevent SCA in highrisk populations and the general community.
]]></description>
<dc:creator>Ashar, F. N.</dc:creator>
<dc:creator>Mitchell, R. N.</dc:creator>
<dc:creator>Albert, C. M.</dc:creator>
<dc:creator>Newton-Cheh, C.</dc:creator>
<dc:creator>Brody, J. A.</dc:creator>
<dc:creator>Muller-Nurasyid, M.</dc:creator>
<dc:creator>Moes, A.</dc:creator>
<dc:creator>Meitinger, T.</dc:creator>
<dc:creator>Mak, A.</dc:creator>
<dc:creator>Huikuri, H.</dc:creator>
<dc:creator>Junttila, M. J.</dc:creator>
<dc:creator>Goyette, P.</dc:creator>
<dc:creator>Pulit, S. L.</dc:creator>
<dc:creator>Pazoki, R.</dc:creator>
<dc:creator>Tanck, M. W.</dc:creator>
<dc:creator>Blom, M. T.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Hauvlinna, A. S.</dc:creator>
<dc:creator>Jabbari, R.</dc:creator>
<dc:creator>Glinge, C.</dc:creator>
<dc:creator>Tragante, V.</dc:creator>
<dc:creator>Escher, S. A.</dc:creator>
<dc:creator>Chakravarti, A.</dc:creator>
<dc:creator>Ehret, G.</dc:creator>
<dc:creator>Coresh, J.</dc:creator>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Prineas, R. J.</dc:creator>
<dc:creator>Franco, O. H.</dc:creator>
<dc:creator>Kwok, P.-Y.</dc:creator>
<dc:creator>Lumley, T.</dc:creator>
<dc:creator>Dumas, F.</dc:creator>
<dc:creator>McKnight, B.</dc:creator>
<dc:creator>Rotter, J. I.</dc:creator>
<dc:creator>Lemaitre, R. N.</dc:creator>
<dc:creator>Heckbert, S. R.</dc:creator>
<dc:creator>O'Donnell, C. J.</dc:creator>
<dc:creator>Hwang, S.-J.</dc:creator>
<dc:creator>Tardif, J.-C.</dc:creator>
<dc:creator>Kortelainen, M.-L.</dc:creator>
<dc:creator>VanDenburgh, M.</dc:creator>
<dc:creator>Uitterlinden, A. G.</dc:creator>
<dc:creator>Hofman,</dc:creator>
<dc:date>2017-12-16</dc:date>
<dc:identifier>doi:10.1101/235234</dc:identifier>
<dc:title><![CDATA[A Comprehensive Evaluation of the Genetic Architecture of Sudden Cardiac Arrest]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/239830v1?rss=1">
<title>
<![CDATA[
Olfactory and Vomeronasal Receptor Feedback Employ Divergent Mechanisms of PERK Activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/239830v1?rss=1</link>
<description><![CDATA[
Mutually-exclusive chemoreceptor expression in olfactory and vomeronasal sensory neurons (OSNs and VSNs) enables odorant discrimination. This configuration involves chemoreceptor mediated activation of the endoplasmic reticulum (ER)-resident kinase PERK. PERK drives translation of the transcription factor ATF5 to preclude additional chemoreceptor expression. ATF5 translation is transient in OSNs but persistent in VSNs, suggesting chemoreceptor-specific modes of PERK activation. Herein, we showed that the ER-lumenal domain (LD) of PERK recognized vomeronasal receptor (VR)-derived peptides, suggesting direct PERK activation drives persistent ATF5 translation in VSNs. In contrast, PERK LD did not recognize olfactory receptor (OR)-derived peptides in vitro, and facilitating OR maturation in vivo prevented PERK activation, suggesting that ORs activate PERK indirectly through a failure to exit the ER. Importantly, impairing or prolonging ATF5 expression drove specific chemoreceptor repertoire biases. Together, these results demonstrate mechanistic divergence in chemoreceptor feedback and establish that differences in PERK activation promote qualitatively different gene regulatory results.
]]></description>
<dc:creator>Dalton, R.</dc:creator>
<dc:creator>Karagoz, E.</dc:creator>
<dc:creator>Kahiapo, J.</dc:creator>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Bashkirova, L.</dc:creator>
<dc:creator>Lyons, D.</dc:creator>
<dc:creator>Matsunami, H.</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:date>2018-01-04</dc:date>
<dc:identifier>doi:10.1101/239830</dc:identifier>
<dc:title><![CDATA[Olfactory and Vomeronasal Receptor Feedback Employ Divergent Mechanisms of PERK Activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/243998v1?rss=1">
<title>
<![CDATA[
Rapid Identification and Validation of Novel Rheumatoid Arthritis Drug Treatments using an Integrative Bioinformatics Platform 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/243998v1?rss=1</link>
<description><![CDATA[
The majority of drugs currently used to treat rheumatoid arthritis (RA) act on a small number of immunomodulatory targets. We applied an integrative biomedical-informatics-based approach and in vivo testing to identify new drug candidates and potential therapeutic targets that could form the basis for future drug development in RA. A computational model of RA was constructed by integrating patient gene expression data, molecular interactions, and clinical drug-disease associations. Drug candidates were scored based on their predicted efficacy across these data types. Ten high-scoring candidates were subsequently screened in a collagen-induced arthritis model of RA. Treatment with exenatide, olopatadine, and TXR-112 significantly improved multiple preclinical endpoints, including animal mobility which was measured using a novel digital platform. These three drug candidates do not act on common RA therapeutic targets; however, links between known candidate pharmacology and pathological processes involved in RA suggest hypothetical mechanisms contributing to the observed efficacy.
]]></description>
<dc:creator>Daugherty, A. C.</dc:creator>
<dc:creator>Farrington, C.</dc:creator>
<dc:creator>Hakin, I.</dc:creator>
<dc:creator>Mujahid, S.</dc:creator>
<dc:creator>Noblin, E. S.</dc:creator>
<dc:creator>Radin, A. M.</dc:creator>
<dc:creator>Chua, M.-S.</dc:creator>
<dc:creator>Rabe, M.</dc:creator>
<dc:creator>Fernald, G.</dc:creator>
<dc:creator>Ford, D.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Schaevitz, L.</dc:creator>
<dc:creator>Radin, A. A.</dc:creator>
<dc:date>2018-01-07</dc:date>
<dc:identifier>doi:10.1101/243998</dc:identifier>
<dc:title><![CDATA[Rapid Identification and Validation of Novel Rheumatoid Arthritis Drug Treatments using an Integrative Bioinformatics Platform]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/244020v1?rss=1">
<title>
<![CDATA[
Real time genetic compensation operationally defines the dynamic demands of feedback control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/244020v1?rss=1</link>
<description><![CDATA[
Biological signaling networks use feedback control to dynamically adjust their operation in real time. Traditional static genetic methods such as gene knockouts or rescue experiments often can identify the existence of feedback interactions, yet are unable to determine what feedback dynamics are required. Here, we implement a new strategy, closed loop optogenetic compensation (CLOC), to address this problem. Using a custom-built hardware and software infrastructure, CLOC monitors in real time the output of a pathway deleted for a feedback regulator. A minimal model uses these measurements to calculate and deliver--on the fly--an optogenetically-enabled transcriptional input designed to compensate for the effects of the feedback deletion. Application of CLOC to the yeast pheromone response pathway revealed surprisingly distinct dynamic requirements for three well-studied feedback regulators. CLOC, a marriage of control theory and traditional genetics, presents a broadly applicable methodology for defining the dynamic function of biological feedback regulators.
]]></description>
<dc:creator>Harrigan, P.</dc:creator>
<dc:creator>Madhani, H.</dc:creator>
<dc:creator>El-Samad, H.</dc:creator>
<dc:date>2018-01-07</dc:date>
<dc:identifier>doi:10.1101/244020</dc:identifier>
<dc:title><![CDATA[Real time genetic compensation operationally defines the dynamic demands of feedback control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/249052v1?rss=1">
<title>
<![CDATA[
Ostreococcus tauri is a high-lipid content green algae that extrudes clustered lipid droplets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/249052v1?rss=1</link>
<description><![CDATA[
Lipid droplet biogenesis, accumulation and secretion is an important field of research spanning biofuel feedstock production in algae and yeast to plant-microbe symbiosis or human metabolic disorders and other diseases. Here we evaluate the critical elements that influence lipid accumulation in the highly simplified and smallest known eukaryote Ostreococcus tauri and identify several conditions that satisfy its classification as an oleaginous green alga. In addition, these experiments revealed the release of excess lipids in pea-pod like structures where many dense lipid droplets are clustered in a linear fashion surrounded by an enveloping membrane which contrasts with known mechanisms from other eukaryotes. These results highlight the potential for Ostreococcus tauri to probe the evolution of lipid droplet dynamics as an emerging model organism with a compacted eukaryotic genome and also to impact lipid feedstock bioproduction applications either directly or using synthetic biology.nnOne Sentence SummaryThe smallest known eukaryote Ostreococcus tauri is oleaginous and sheds lipid droplets as pea-pod like membrane enclosed clusters.
]]></description>
<dc:creator>Smallwood, C. R.</dc:creator>
<dc:creator>Chrisler, W.</dc:creator>
<dc:creator>Chen, J.-H.</dc:creator>
<dc:creator>Patello, E.</dc:creator>
<dc:creator>Thomas, M.</dc:creator>
<dc:creator>Boudreau, R.</dc:creator>
<dc:creator>Ekman, A.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>McDermott, G.</dc:creator>
<dc:creator>Evans, J. E.</dc:creator>
<dc:date>2018-01-17</dc:date>
<dc:identifier>doi:10.1101/249052</dc:identifier>
<dc:title><![CDATA[Ostreococcus tauri is a high-lipid content green algae that extrudes clustered lipid droplets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/252031v1?rss=1">
<title>
<![CDATA[
Next generation CRISPR/Cas9 transcriptional activation in Drosophila using flySAM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/252031v1?rss=1</link>
<description><![CDATA[
CRISPR/Cas9-based transcriptional activation (CRISPRa) has recently emerged as a powerful and scalable technique for systematic over-expression genetic analysis in Drosophila melanogaster. We present flySAM, a potent new tool for in vivo CRISPRa, which offers a major improvement over existing strategies in terms of effectiveness, scalability, and ease-of-use. flySAM outperforms existing in vivo CRISPRa strategies, and approximates phenotypes obtained using traditional Gal4-UAS over-expression. Further, because flySAM typically only requires a single sgRNA, it dramatically improves scalability. We use flySAM to demonstrate multiplexed CRISPRa, which has not been previously shown in vivo. In addition, we have simplified the experimental usage of flySAM by creating a single vector encoding both the UAS:Cas9-activator and the sgRNA, allowing for inducible CRISPRa in a single genetic cross. flySAM will thus replace previous CRISPRa strategies as the basis of our growing genome-wide transgenic over-expression resource, TRiP-OE.
]]></description>
<dc:creator>Jia, Y.</dc:creator>
<dc:creator>Xu, R.-G.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Ewen-Campen, B.</dc:creator>
<dc:creator>Rajakumar, R.</dc:creator>
<dc:creator>Zirin, J.</dc:creator>
<dc:creator>Yang-Zhou, D.</dc:creator>
<dc:creator>Zhao, R.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Mao, D.</dc:creator>
<dc:creator>Peng, P.</dc:creator>
<dc:creator>Qiao, H.-H.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Liu, L.-P.</dc:creator>
<dc:creator>Xu, B.</dc:creator>
<dc:creator>Ji, J.-Y.</dc:creator>
<dc:creator>Lu, Q.</dc:creator>
<dc:creator>Sun, J.</dc:creator>
<dc:creator>Perrimon, N.</dc:creator>
<dc:creator>Ni, J.-Q.</dc:creator>
<dc:date>2018-01-31</dc:date>
<dc:identifier>doi:10.1101/252031</dc:identifier>
<dc:title><![CDATA[Next generation CRISPR/Cas9 transcriptional activation in Drosophila using flySAM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/253872v1?rss=1">
<title>
<![CDATA[
OGT binds a conserved C-terminal domain of TET1 toregulate TET1 activity and function in development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/253872v1?rss=1</link>
<description><![CDATA[
TET enzymes convert 5-methylcytosine to 5-hydroxymethylcytosine and higher oxidized derivatives. TETs stably associate with and are post-translationally modified by the nutrient-sensing enzyme OGT, suggesting a connection between metabolism and the epigenome. Here, we show for the first time that modification by OGT enhances TET1 activity in vitro. We identify a domain of TET1 responsible for binding to OGT and report a point mutation that disrupts the TET1-OGT interaction. We show that the TET1-OGT interaction is necessary for TET1 to rescue hematopoetic stem cell production in tet mutant zebrafish embryos, suggesting that OGT promotes TET1s function during development. Finally, we show that disrupting the TET1-OGT interaction in mouse embryonic stem cells changes the abundance of TET-containing high molecular weight complexes and causes widespread gene expression changes. These results link metabolism and epigenetic control, which may be relevant to the developmental and disease processes regulated by these two enzymes.
]]></description>
<dc:creator>Hrit, J.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Martin, E. A.</dc:creator>
<dc:creator>Simental, E.</dc:creator>
<dc:creator>Goll, M.</dc:creator>
<dc:creator>Panning, B.</dc:creator>
<dc:date>2018-01-25</dc:date>
<dc:identifier>doi:10.1101/253872</dc:identifier>
<dc:title><![CDATA[OGT binds a conserved C-terminal domain of TET1 toregulate TET1 activity and function in development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/263731v1?rss=1">
<title>
<![CDATA[
Conserved Roles of C. elegans and Human MANFs in Sulfatide Binding and Cytoprotection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/263731v1?rss=1</link>
<description><![CDATA[
Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF) is an endoplasmic reticulum (ER) protein that can be secreted and protect dopamine neurons and cardiomyocytes from ER stress and apoptosis. The mechanism of action of extracellular MANF has long been elusive. From a genetic screen for mutants with abnormal ER stress response, we identified the gene Y54G2A.23 as the evolutionarily conserved C. elegans MANF orthologue. We find that MANF binds to the lipid sulfatide, also known as 3-O-sulfogalactosylceramide present in serum and outer-cell membrane leaflets, directly in isolated forms and in reconstituted lipid micelles. Sulfatide binding promotes cellular MANF uptake and cytoprotection from hypoxia-induced cell death. Heightened ER stress responses of MANF-null C. elegans mutants and mammalian cells are alleviated by human MANF in a sulfatide-dependent manner. Our results demonstrate conserved roles of MANF in sulfatide binding and ER stress response, supporting sulfatide as a long-sought lipid mediator of MANFs cytoprotection.
]]></description>
<dc:creator>Bai, M.</dc:creator>
<dc:creator>Vozdek, R.</dc:creator>
<dc:creator>Hnizda, A.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Kuchar, L.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Wood, C.</dc:creator>
<dc:creator>Feng, L.</dc:creator>
<dc:creator>Dang, Y.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2018-02-11</dc:date>
<dc:identifier>doi:10.1101/263731</dc:identifier>
<dc:title><![CDATA[Conserved Roles of C. elegans and Human MANFs in Sulfatide Binding and Cytoprotection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/269522v1?rss=1">
<title>
<![CDATA[
SOX2 Regulation by Hedgehog Signaling Controls Adult Lingual Epithelium Homeostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/269522v1?rss=1</link>
<description><![CDATA[
The adult tongue epithelium is continuously renewed from epithelial progenitor cells, and this process relies on intact Hedgehog (HH) signaling. In mice, inhibition of the HH pathway using Smoothened antagonists (HH pathway inhibitors or HPIs) leads to taste bud loss over a span of several weeks. Previously, we demonstrated that overexpression of Sonic Hedgehog (SHH) in lingual epithelial progenitors induces formation of ectopic taste buds accompanied by locally increased SOX2 expression, consistent with the hypothesis that taste bud differentiation depends on SOX2 downstream of HH. To test this idea, we inhibited HH signaling by treating SOX2-GFP mice with HPI and found a rapid and drastic decline in SOX2-GFP expression in taste progenitors and taste buds. Using a conditional Cre-lox system to delete Sox2, we found that loss of SOX2 blocks differentiation of both taste buds and non-taste epithelium that comprises the majority of the tongue surface; progenitor cells increase in number at the expense of differentiated taste cells and lingual keratinocytes. In contrast to the normal pattern of basally restricted proliferation, dividing cells are overabundant, disorganized and present in suprabasal epithelial layers in Sox2 deleted tongues. Additionally, SOX2 loss in taste progenitors leads non-cell autonomously to rapid loss of taste bud cells via apoptosis, dramatically shortening taste cell lifespans. Finally, when Sox2 is conditionally deleted in mice with constitutive overexpression of SHH, ectopic taste buds fail to form and endogenous taste buds disappear; instead, robust hyperproliferation takes over the entire lingual epithelium. In sum, our experiments suggest that SOX2 functions downstream of HH signaling to regulate lingual epithelium homeostasis.
]]></description>
<dc:creator>Castillo-Azofeifa, D.</dc:creator>
<dc:creator>Seidel, K.</dc:creator>
<dc:creator>Gross, L.</dc:creator>
<dc:creator>Jacquez, B.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Barlow, L. A.</dc:creator>
<dc:date>2018-02-22</dc:date>
<dc:identifier>doi:10.1101/269522</dc:identifier>
<dc:title><![CDATA[SOX2 Regulation by Hedgehog Signaling Controls Adult Lingual Epithelium Homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/271106v1?rss=1">
<title>
<![CDATA[
A Genetic Program Mediates Cold-warming Response and Promotes Stress-induced Phenoptosis in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/271106v1?rss=1</link>
<description><![CDATA[
How multicellular organisms respond to and are impacted by severe hypothermic stress is largely unknown. From C. elegans screens for mutants abnormally responding to cold-warming stimuli, we identify a molecular genetic pathway comprising ISY-1, a conserved uncharacterized protein, and ZIP-10, a bZIP-type transcription factor. ISY-1 gatekeeps the ZIP-10 transcriptional program by regulating the microRNA mir-60. Downstream of ISY-1 and mir-60, zip-10 levels rapidly and specifically increase upon transient cold-warming response. Prolonged zip-10 up-regulation induces several protease-encoding genes and promotes stress-induced organismic death, or phenoptosis, of C. elegans. zip-10 deficiency confers enhanced resistance to prolonged cold-warming stress, more prominently in adults than larvae. We conclude that the ZIP-10 genetic program mediates cold-warming response and may have evolved to promote wild population kin selection under resource-limiting and thermal stress conditions.
]]></description>
<dc:creator>Jiang, W.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Long, Y.</dc:creator>
<dc:creator>Owen, A.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Luo, S.</dc:creator>
<dc:creator>Dang, Y.</dc:creator>
<dc:creator>Ma, D.</dc:creator>
<dc:date>2018-02-24</dc:date>
<dc:identifier>doi:10.1101/271106</dc:identifier>
<dc:title><![CDATA[A Genetic Program Mediates Cold-warming Response and Promotes Stress-induced Phenoptosis in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/272328v1?rss=1">
<title>
<![CDATA[
A novel xenograft model reveals invasive mesenchymal transition and ineffective angiogenic response during the evolution of resistance to anti-angiogenic therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/272328v1?rss=1</link>
<description><![CDATA[
Bevacizumab treatment of glioblastoma is limited by transient responses and acquired resistance. Because of the lengthy duration of treatment that can precede resistance in patients, in order to study changes underlying the evolution of bevacizumab resistance, we created a novel multigenerational xenograft model of acquired bevacizumab resistance. Glioblastoma xenografts were treated with bevacizumab or IgG, and the fastest growing tumor re-implanted into new mice, generating paired isogeneic responsive or resistant multigenerational xenografts. Microarray analysis revealed significant overexpression across generations of the mesenchymal subtype gene signature, paralleling results from patient bevacizumab-resistant glioblastomas (BRGs) that exhibited increasing mesenchymal gene expression correlating with increased bevacizumab treatment duration. Key mesenchymal markers, including YKL-40, CD44, SERPINE1, and TIMP1 were upregulated across generations, with altered morphology, increased invasiveness, and increased neurosphere formation confirmed in later xenograft generations. Interestingly, YKL-40 levels were elevated in serum of patients with bevacizumab-resistant vs. bevacizumab-naive glioblastomas. Finally, despite upregulation of VEGF-independent pro-angiogenic genes across xenograft generations, immunostaining revealed increased hypoxia and decreased vessel density with increasing generation of treatment, mirroring our findings in patient BRGs and suggesting tumor growth despite effective devascularization caused by VEGF blockade. Besides identifying novel targets for preventing the evolution of resistance and offering a xenograft model for testing resistance inhibitors, our work suggests YKL-40 as a blood biomarker of bevacizumab resistance worthy of further evaluation.
]]></description>
<dc:creator>Jahangiri, A.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Chandra, A.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Weiss, J.</dc:creator>
<dc:creator>Wolf, K.</dc:creator>
<dc:creator>Lin, G.</dc:creator>
<dc:creator>Rick, J.</dc:creator>
<dc:creator>Sidorov, M.</dc:creator>
<dc:creator>Flanigan, P.</dc:creator>
<dc:creator>Yagnik, G.</dc:creator>
<dc:creator>Carbonell, W. S.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:date>2018-02-28</dc:date>
<dc:identifier>doi:10.1101/272328</dc:identifier>
<dc:title><![CDATA[A novel xenograft model reveals invasive mesenchymal transition and ineffective angiogenic response during the evolution of resistance to anti-angiogenic therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/278556v1?rss=1">
<title>
<![CDATA[
Robotic automation of droplet microfluidics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/278556v1?rss=1</link>
<description><![CDATA[
Droplet microfluidics enables new reactions, assays, and analytic capabilities, but often requires complex workflows involving numerous steps of macro- and micro-fluidic processing. We demonstrate robotically-automated droplet microfluidics, an approach to automate workflows with commercial fluid-handling robots. These workflows can be performed without human intervention, increasing reliability and convenience.
]]></description>
<dc:creator>Tran, T. M.</dc:creator>
<dc:creator>Kim, S. C.</dc:creator>
<dc:creator>Abate, A. R.</dc:creator>
<dc:date>2018-03-07</dc:date>
<dc:identifier>doi:10.1101/278556</dc:identifier>
<dc:title><![CDATA[Robotic automation of droplet microfluidics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/284604v1?rss=1">
<title>
<![CDATA[
Single-cell RNA-seq reveals profibrotic macrophages in lung fibrosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/284604v1?rss=1</link>
<description><![CDATA[
Myeloid cells localize to peripheral tissues in a wide range of pathologic contexts. However, appreciation of distinct myeloid subtypes has been limited by the signal averaging inherent to bulk sequencing approaches. Here we applied single-cell RNA sequencing (scRNA-seq) to map cellular heterogeneity in lung fibrosis induced by bleomycin injury in mice. We first developed a computational framework that enables unbiased, granular cell-type annotation of scRNA-seq. This approach identified a macrophage subpopulation that was specific to injured lung and notable for high expression of Cx3cr1+ and MHCII genes. We found that these macrophages, which bear a gene expression profile consistent with monocytic origin, progressively acquire alveolar macrophage identity and localize to sites of fibroblast accumulation. Probing their functional role, in vitro studies showed a trophic effect of these cells on fibroblast activation, and ablation of Cx3cr1-expressing cells suppressed fibrosis in vivo. We also found by gene set analysis and immunofluorescence that markers of these macrophages were upregulated in samples from patients with lung fibrosis compared with healthy controls. Taken together, our results uncover a specific pathologic subgroup of macrophages with markers that could enable their therapeutic targeting for fibrosis.
]]></description>
<dc:creator>Aran, D.</dc:creator>
<dc:creator>Looney, A. P.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Fong, V.</dc:creator>
<dc:creator>Hsu, A.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:creator>Abate, A.</dc:creator>
<dc:creator>Butte, A. J.</dc:creator>
<dc:creator>Bhattacharya, M.</dc:creator>
<dc:date>2018-03-19</dc:date>
<dc:identifier>doi:10.1101/284604</dc:identifier>
<dc:title><![CDATA[Single-cell RNA-seq reveals profibrotic macrophages in lung fibrosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/284992v1?rss=1">
<title>
<![CDATA[
Avian keratin disorder of Alaska black-capped chickadees is associated with Poecivirus infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/284992v1?rss=1</link>
<description><![CDATA[
BackgroundAvian keratin disorder (AKD) is an epizootic of debilitating beak deformities, first documented in black-capped chickadees (Poecile atricapillus) in Alaska during the late 1990s. Similar deformities have now been recorded in dozens of species of birds across multiple continents. Despite this, the etiology of AKD has remained elusive, making it difficult to assess the impacts of this disease on wild populations. We previously identified an association between infection with a novel picornavirus, Poecivirus, and AKD in a small cohort of black-capped chickadees.nnMethodsTo test if the association between Poecivirus and AKD holds in a lager study population, we used targeted PCR followed by Sanger sequencing to screen 124 symptomatic and asymptomatic black-capped chickadees for Poecivirus infection. We further compared the efficacy of multiple non-terminal field sampling methods (buccal swabs, cloacal swabs, fecal samples, and blood samples) for Poecivirus screening. Finally, we used both in situ hybridization and a strand-specific expression assay to localize Poecivirus to beak tissue of AKD-positive individuals and to determine if virus is actively replicating in beak tissue.nnResultsPoecivirus was detected in 29/29 (100%) individuals with AKD, but only 9/95 (9.5%) asymptomatic individuals with apparently normal beaks (p < 0.0001). We found that cloacal swabs are the most sensitive of these sample types for detecting Poecivirus in birds with AKD, but that buccal swabs should be combined with cloacal swabs in evaluating the infection status of asymptomatic birds. Finally, we used both in situ hybridization and a strand-specific expression assay to localize Poecivirus to beak tissue of AKD-positive individuals and to provide evidence of active viral replication.nnConclusionThe data presented here show a strong, statistically significant relationship between Poecivirus infection and AKD, and provide evidence that Poecivirus is indeed an avian virus, infecting and actively replicating in beak tissue of AKD-affected BCCH. Taken together, these data corroborate and extend the evidence for a potential causal association between Poecivirus and AKD in the black-capped chickadee. Poecivirus continues to warrant further investigation as a candidate agent of AKD.
]]></description>
<dc:creator>Zylberberg, M.</dc:creator>
<dc:creator>Van Hemert, C.</dc:creator>
<dc:creator>Handel, C. M.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:date>2018-03-19</dc:date>
<dc:identifier>doi:10.1101/284992</dc:identifier>
<dc:title><![CDATA[Avian keratin disorder of Alaska black-capped chickadees is associated with Poecivirus infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/295501v1?rss=1">
<title>
<![CDATA[
Diverse progenitor cells preserve salivary gland ductal architecture after radiation induced damage 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/295501v1?rss=1</link>
<description><![CDATA[
The ductal system of the salivary gland has long been postulated to be resistant to radiation-induced damage, a common outcome incurred by head and neck cancer patients receiving radiotherapy. Yet, whether the ducts are capable of regenerating after genotoxic injury, or if damage to ductal cells induces lineage plasticity, as has been reported in other organ systems, remains unknown. Here, we show that two ductal progenitor populations marked by KRT14 and KIT exclusively maintain non-overlapping ductal compartments after radiation exposure but do so through distinct cellular mechanisms. KRT14+ progenitor cells are fast cycling cells that proliferate in response to radiation-induced damage in a sustained manner and divide asymmetrically to produce differentiated cells of the larger granulated ducts. Conversely, KIT+ cells are long lived progenitors for the intercalated ducts that undergo few cell divisions either during homeostasis or after gamma radiation, thus maintaining ductal architecture in the near absence of cell turnover. Together, these data illustrate the regenerative capacity of the salivary ducts and highlight the heterogeneity in the damage responses used by salivary progenitor cells to maintain tissue architecture.nnSummary StatementThe salivary gland ductal network is maintained during homeostasis and after genotoxic injury by diverse progenitors that respond differentially to radiation induced damage.
]]></description>
<dc:creator>May, A.</dc:creator>
<dc:creator>Cruz Pacheco, N.</dc:creator>
<dc:creator>Emmerson, E.</dc:creator>
<dc:creator>Siedel, K.</dc:creator>
<dc:creator>Nathan, S.</dc:creator>
<dc:creator>Muench, M. O.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Knox, S. M.</dc:creator>
<dc:date>2018-04-05</dc:date>
<dc:identifier>doi:10.1101/295501</dc:identifier>
<dc:title><![CDATA[Diverse progenitor cells preserve salivary gland ductal architecture after radiation induced damage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/298349v1?rss=1">
<title>
<![CDATA[
Approaches to maximize sgRNA-barcode coupling in Perturb-seq screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/298349v1?rss=1</link>
<description><![CDATA[
Perturb-seq is a platform for single-cell gene expression profiling of pooled CRISPR screens. Like many functional genomics platforms, Perturb-seq relies on lentiviral transduction to introduce perturbation libraries to cells. On this platform, these are barcoded sgRNA libraries. A critical consideration for performing Perturb-seq experiments is uncoupling of barcodes from linked sgRNA expression cassettes, which can occur during lentiviral transduction of co-packaged libraries due to reverse transcriptase-mediated template switching. This problem is common to lentiviral libraries designed with linked variable regions. Here, we demonstrate that recombination between Perturb-seq vectors scrambles linked variable regions separated by 2 kb. This predicts information loss in Perturb-seq screens performed with co-packaged libraries. We also demonstrate ways to address this problem and discuss best practices for single-cell screens with transcriptional readouts.
]]></description>
<dc:creator>Adamson, B.</dc:creator>
<dc:creator>Norman, T. M.</dc:creator>
<dc:creator>Jost, M.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:date>2018-04-11</dc:date>
<dc:identifier>doi:10.1101/298349</dc:identifier>
<dc:title><![CDATA[Approaches to maximize sgRNA-barcode coupling in Perturb-seq screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/309682v1?rss=1">
<title>
<![CDATA[
Construction of a novel phagemid to produce custom DNA origami scaffolds 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/309682v1?rss=1</link>
<description><![CDATA[
DNA origami, a method for constructing nanoscale objects, relies on a long single strand of DNA to act as the "scaffold" to template assembly of numerous short DNA oligonucleotide "staples". The ability to generate custom scaffold sequences can greatly benefit DNA origami design processes. Custom scaffold sequences can provide better control of the overall size of the final object and better control of low-level structural details, such as locations of specific base pairs within an object. Filamentous bacteriophages and related phagemids can work well as sources of custom scaffold DNA. However, scaffolds derived from phages require inclusion of multi-kilobase DNA sequences in order to grow in host bacteria, and thus cannot be altered or removed. These fixed-sequence regions constrain the design possibilities of DNA origami. Here we report the construction of a novel phagemid, pScaf, to produce scaffolds that have a custom sequence with a much smaller fixed region of only 381 bases. We used pScaf to generate new scaffolds ranging in size from 1,512 to 10,080 bases and demonstrated their use in various DNA origami shapes and assemblies. We anticipate our pScaf phagemid will enhance development of the DNA origami method and its future applications.
]]></description>
<dc:creator>Nafisi, P. M.</dc:creator>
<dc:creator>Aksel, T.</dc:creator>
<dc:creator>Douglas, S. M.</dc:creator>
<dc:date>2018-04-27</dc:date>
<dc:identifier>doi:10.1101/309682</dc:identifier>
<dc:title><![CDATA[Construction of a novel phagemid to produce custom DNA origami scaffolds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/311498v1?rss=1">
<title>
<![CDATA[
Mechanistic Origins of Dynamic Instability in Filaments from the Phage Tubulin, PhuZ 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/311498v1?rss=1</link>
<description><![CDATA[
A bacteriophage-encoded tubulin homologue, PhuZ, harnesses dynamic instability to position genomes of KZ-like bacteriophage at the midline of their Pseudomonas hosts, facilitating phage infectivity. While much has been learned about molecular origins of microtubule dynamics, how GTP binding and hydrolysis control dynamics in the divergent 3-stranded PhuZ filaments is not understood. Here we present cryo-EM reconstructions of the PhuZ filamentin a pre-hydrolysis (3.5[A]) and three post-hydrolysis states (4.2 [A], 7.3 [A] and 8.1 [A] resolutions), likely representing distinct depolymerization stages. Core polymerization-induced structural changes reveal similarities to {beta}-tubulin, suggesting broad conservation within the tubulin family. By contrast, GTP hydrolysis is sensed quite differently and is communicated by the divergent PhuZ C-terminus to the lateral interface, leading to PhuZ polymer destabilization. This provides a contrasting molecular description of how nucleotide state can be harnessed by the tubulin fold to regulate filament assembly, metastability and disassembly.
]]></description>
<dc:creator>Zehr, E. A.</dc:creator>
<dc:creator>Rohu, A.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Verba, K. A.</dc:creator>
<dc:creator>Pogliano, J.</dc:creator>
<dc:creator>Grigorieff, N.</dc:creator>
<dc:creator>Agard, D. A.</dc:creator>
<dc:date>2018-04-30</dc:date>
<dc:identifier>doi:10.1101/311498</dc:identifier>
<dc:title><![CDATA[Mechanistic Origins of Dynamic Instability in Filaments from the Phage Tubulin, PhuZ]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/312611v1?rss=1">
<title>
<![CDATA[
Heart enhancers with deeply conserved regulatory activity are established early in development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/312611v1?rss=1</link>
<description><![CDATA[
During the phylotypic period embryos from different genera show similar gene expression patterns, implying common regulatory mechanisms. To identify enhancers involved in the initial events of cardiogenesis, which occurs during the phylotypic period, we isolated early cardiac progenitor cells from zebrafish embryos and characterized 3838 open chromatin regions specific to this cell population. Of these regions, 162 overlapped with conserved non-coding elements (CNEs) that also mapped to open chromatin regions in human. Most of the zebrafish conserved open chromatin elements tested drove gene expression in the developing heart. Despite modest sequence identity, human orthologous open chromatin regions could recapitulate the spatial temporal expression patterns of the zebrafish sequence, potentially providing a basis for phylotypic gene expression patterns. Genome-wide, we discovered 5598 zebrafish-human conserved open chromatin regions, suggesting that a diverse repertoire of ancient enhancers is established prior to organogenesis and the phylotypic period.
]]></description>
<dc:creator>Yuan, X.</dc:creator>
<dc:creator>Song, M.</dc:creator>
<dc:creator>Devine, P.</dc:creator>
<dc:creator>Bruneau, B. G.</dc:creator>
<dc:creator>Scott, I. C.</dc:creator>
<dc:creator>Wilson, M. D.</dc:creator>
<dc:date>2018-05-04</dc:date>
<dc:identifier>doi:10.1101/312611</dc:identifier>
<dc:title><![CDATA[Heart enhancers with deeply conserved regulatory activity are established early in development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/314344v1?rss=1">
<title>
<![CDATA[
crisprQTL mapping as a genome-wide association framework for cellular genetic screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/314344v1?rss=1</link>
<description><![CDATA[
Expression quantitative trait locus (eQTL) and genome-wide association studies (GWAS) are powerful paradigms for mapping the determinants of gene expression and organismal phenotypes, respectively. However, eQTL mapping and GWAS are limited in scope (to naturally occurring, common genetic variants) and resolution (by linkage disequilibrium). Here, we present crisprQTL mapping, a framework in which large numbers of CRISPR/Cas9 perturbations are introduced to each cell on an isogenic background, followed by single-cell RNA-seq (scRNA-seq). crisprQTL mapping is analogous to conventional human eQTL studies, but with individual humans replaced by individual cells; genetic variants replaced by unique combinations of  unlinked guide RNA (gRNA)-programmed perturbations per cell; and tissue-level RNA-seq of many individuals replaced by scRNA-seq of many cells. By randomly introducing gRNAs, a single population of cells can be leveraged to test for association between each perturbation and the expression of any potential target gene, analogous to how eQTL studies leverage populations of humans to test millions of genetic variants for associations with expression in a genome-wide manner. However, crisprQTL mapping is neither limited to naturally occurring, common genetic variants nor by linkage disequilibrium. As a proof-of-concept, we applied crisprQTL mapping to evaluate 1,119 candidate enhancers with no strong a priori hypothesis as to their target gene(s). Perturbations were made by a nuclease-dead Cas9 (dCas9) tethered to KRAB, and introduced at a mean  allele frequency of 1.1% into a population of 47,650 profiled human K562 cells (median of 15 gRNAs identified per cell). We tested for differential expression of all genes within 1 megabase (Mb) of each candidate enhancer, effectively evaluating 17,584 potential enhancer-target gene relationships within a single experiment. At an empirical false discovery rate (FDR) of 10%, we identify 128 cis crisprQTLs (11%) whose targeting resulted in downregulation of 105 nearby genes. crisprQTLs were strongly enriched for proximity to their target genes (median 34.3 kilobases (Kb)) and the strength of H3K27ac, p300, and lineage-specific transcription factor (TF) ChIP-seq peaks. Our results establish the power of the eQTL mapping paradigm as applied to programmed variation in populations of cells, rather than natural variation in populations of individuals. We anticipate that crisprQTL mapping will facilitate the comprehensive elucidation of the cis-regulatory architecture of the human genome.
]]></description>
<dc:creator>Gasperini, M.</dc:creator>
<dc:creator>Hill, A.</dc:creator>
<dc:creator>McFaline-Figueroa, J. L.</dc:creator>
<dc:creator>Martin, B.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:date>2018-05-04</dc:date>
<dc:identifier>doi:10.1101/314344</dc:identifier>
<dc:title><![CDATA[crisprQTL mapping as a genome-wide association framework for cellular genetic screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/316323v1?rss=1">
<title>
<![CDATA[
Chimeric antigen receptors that trigger phagocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/316323v1?rss=1</link>
<description><![CDATA[
Chimeric antigen receptors (CARs) are synthetic receptors that reprogram T cells to kill cancer. The success of CAR-T cell therapies highlights the promise of programmed immunity, and suggests that applying CAR strategies to other immune cell lineages may be beneficial. Here, we engineered a family of Chimeric Antigen Receptors for Phagocytosis (CAR-Ps) that direct macrophages to engulf specific targets, including cancer cells. CAR-Ps consist of an extracellular antibody fragment, which can be modified to direct CAR-P activity towards specific antigens. By screening a panel of engulfment receptor intracellular domains, we found that the cytosolic domains from Megf10 and FcR{gamma} robustly triggered engulfment independently of their native extracellular domain. We show that CAR-Ps drive specific engulfment of antigen-coated synthetic particles and whole cancer cells. Addition of a tandem PI3K recruitment domain increased cancer cell engulfment. Finally, we show that CAR-P expressing macrophages reduce cancer cell number in co-culture by over 40%.nnSummaryWe report the first Chimeric Antigen Receptors for Phagocytosis (CAR-Ps) that promote engulfment of antigen-coated particles and cancer cells.
]]></description>
<dc:creator>Morrissey, M. A.</dc:creator>
<dc:creator>Williamson, A. P.</dc:creator>
<dc:creator>Steinbach, A. M.</dc:creator>
<dc:creator>Roberts, E. W.</dc:creator>
<dc:creator>Kern, N.</dc:creator>
<dc:creator>Headley, M. B.</dc:creator>
<dc:creator>Vale, R. D.</dc:creator>
<dc:date>2018-05-07</dc:date>
<dc:identifier>doi:10.1101/316323</dc:identifier>
<dc:title><![CDATA[Chimeric antigen receptors that trigger phagocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/316679v1?rss=1">
<title>
<![CDATA[
Decoupling the impact of microRNAs on translational repression versus RNA degradation in embryonic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/316679v1?rss=1</link>
<description><![CDATA[
Translation and mRNA degradation are intimately connected, yet the mechanisms that regulate them are not fully understood. Here we studied the link between translation and mRNA stability in embryonic stem cells (ESCs). Transcripts showed a wide range of stabilities, which correlated with their translation levels. The protein DHH1 links translation to mRNA stability in yeast; however loss of the mammalian homolog, DDX6, in ESCs did not disrupt the correlation across transcripts. Instead, the loss of DDX6 led to upregulated translation of microRNA targets, without concurrent changes in mRNA stability. The Ddx6 knockout cells were phenotypically and molecularly similar to cells lacking all microRNAs (Dgcr8 knockout ESCs). These data show that the loss of DDX6 can separate the two canonical functions of microRNAs: translational repression and transcript destabilization. Furthermore, these data uncover a central role for translational repression independent of transcript destabilization in defining the downstream consequences of microRNA loss.
]]></description>
<dc:creator>Blelloch, R.</dc:creator>
<dc:creator>Freimer, J. W.</dc:creator>
<dc:creator>Hu, T.</dc:creator>
<dc:date>2018-05-07</dc:date>
<dc:identifier>doi:10.1101/316679</dc:identifier>
<dc:title><![CDATA[Decoupling the impact of microRNAs on translational repression versus RNA degradation in embryonic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/317800v1?rss=1">
<title>
<![CDATA[
The Clinical Imperative for Inclusivity: Race, Ethnicity, and Ancestry (REA) in Genomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/317800v1?rss=1</link>
<description><![CDATA[
The Clinical Genome Resource (ClinGen) Ancestry and Diversity Working Group highlights the need to develop guidance on race, ethnicity, and ancestry (REA) data collection and use in clinical genomics. We present quantitative and qualitative evidence to characterize: 1) acquisition of REA data via clinical laboratory requisition forms, and 2) information disparity across populations in the Genome Aggregation Database (gnomAD) at clinically relevant sites as determined by variants in ClinVar. Our requisition form analysis showed substantial heterogeneity in clinical laboratory ascertainment of REA, as well as marked incongruity among terms used to define REA categories. There was also striking disparity across REA populations in the amount of information available about variants at clinically relevant sites in gnomAD. European ancestral populations constituted the majority of observations (55.8%), allele counts (59.7%), and private alleles (56.1%) in gnomAD at 550 loci with "pathogenic" and "likely pathogenic" expert-reviewed variants in ClinVar. Our findings highlight the importance of implementing and supporting programs to increase diversity in genome sequencing and clinical genomics, as well as measuring uncertainty around population-level datasets that are used in variant interpretation. Finally, we suggest the need for a standardized REA data collection framework to be developed and adopted across clinical genomics.
]]></description>
<dc:creator>Popejoy, A. B.</dc:creator>
<dc:creator>Ritter, D. I.</dc:creator>
<dc:creator>Crooks, K.</dc:creator>
<dc:creator>Currey, E.</dc:creator>
<dc:creator>Fullerton, S. M.</dc:creator>
<dc:creator>Hindorff, L. A.</dc:creator>
<dc:creator>Koenig, B.</dc:creator>
<dc:creator>Ramos, E. M.</dc:creator>
<dc:creator>Sorokin, E. P.</dc:creator>
<dc:creator>Wand, H.</dc:creator>
<dc:creator>Wright, M. W.</dc:creator>
<dc:creator>Zou, J.</dc:creator>
<dc:creator>Gignoux, C. R.</dc:creator>
<dc:creator>Bonham, V. L.</dc:creator>
<dc:creator>Plon, S. E.</dc:creator>
<dc:creator>Bustamante, C. D.</dc:creator>
<dc:creator>The Clinical Genome Resource (ClinGen) Ancestry and Diversity Working Group (ADWG),</dc:creator>
<dc:date>2018-05-09</dc:date>
<dc:identifier>doi:10.1101/317800</dc:identifier>
<dc:title><![CDATA[The Clinical Imperative for Inclusivity: Race, Ethnicity, and Ancestry (REA) in Genomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/327478v1?rss=1">
<title>
<![CDATA[
A data mining paradigm for identifying key factors in biological processes using gene expression data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/327478v1?rss=1</link>
<description><![CDATA[
A large volume of biological data is being generated for studying mechanisms of various biological processes. These precious data enable large-scale computational analyses to gain biological insights. However, it remains a challenge to mine the data efficiently for knowledge discovery. The heterogeneity of these data makes it difficult to consistently integrate them, slowing down the process of biological discovery. We introduce a data processing paradigm to identify key factors in biological processes via systematic collection of gene expression datasets, primary analysis of data, and evaluation of consistent signals. To demonstrate its effectiveness, our paradigm was applied to epidermal development and identified many genes that play a potential role in this process. Besides the known epidermal development genes, a substantial proportion of the identified genes are still not supported by gain- or loss-of-function studies, yielding many novel genes for future studies. Among them, we selected a top gene for loss-of-function experimental validation and confirmed its function in epidermal differentiation, proving the ability of this paradigm to identify new factors in biological processes. In addition, this paradigm revealed many key genes in cold-induced thermogenesis using data from cold-challenged tissues, demonstrating its generalizability. This paradigm can lead to fruitful results for studying molecular mechanisms in an era of explosive accumulation of publicly available biological data.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Zheng, L.</dc:creator>
<dc:creator>Uchiyama, A.</dc:creator>
<dc:creator>Bin, L.</dc:creator>
<dc:creator>Mauro, T. M.</dc:creator>
<dc:creator>Elias, P. M.</dc:creator>
<dc:creator>Pawelczyk, T.</dc:creator>
<dc:creator>Sakowicz-Burkiewicz, M.</dc:creator>
<dc:creator>Trzeciak, M.</dc:creator>
<dc:creator>Leung, D. Y. M.</dc:creator>
<dc:creator>Morasso, M. I.</dc:creator>
<dc:creator>Yu, P.</dc:creator>
<dc:date>2018-05-21</dc:date>
<dc:identifier>doi:10.1101/327478</dc:identifier>
<dc:title><![CDATA[A data mining paradigm for identifying key factors in biological processes using gene expression data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/331082v1?rss=1">
<title>
<![CDATA[
Cofilin-actin rod formation in neuronal processes after brain ischemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/331082v1?rss=1</link>
<description><![CDATA[
Functional impairment after brain ischemia results in part from loss of neuronal spines and dendrites, independent of neuronal death. Cofilin-actin rods are covalently linked aggregates of colfilin-1 and actin that form in neuronal processes (neurites) under conditions of ATP depletion and oxidative stress, and which cause neurite degeneration if not disassembled. ATP depletion and oxidative stress occur with differing severity, duration, and time course in different ischemic conditions. Here we evaluated four mouse models of brain ischemia to define the conditions that drive formation of cofilin-actin rods. Three of the models provide early reperfusion: transient middle cerebral artery occlusion (MCAo), transient bilateral common carotid occlusion (CCAo), and cardiac arrest / cardiopulmonary resuscitation (CA/CPR). Early reperfusion restores ATP generating capacity, but also induces oxidative stress. The fourth model, photothrombotic cortical infarction, does not provide reperfusion. Cofilin-actin rods were formed in each of these models, but with differing patterns. Where acute reperfusion occurred, rod formation was maximal within 4 hours after reperfusion. Where infarction occurred, rods continued to form for at least 24 hours after ischemic onset, and extended into the adjacent non-ischemic tissue. Interventions that limit cofilin-actin rod formation may help to preserve integrity of neuronal processes in permanent ischemia.
]]></description>
<dc:creator>Won, S. J.</dc:creator>
<dc:creator>Minnella, A. M.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Eun, C. H.</dc:creator>
<dc:creator>Rome, E.</dc:creator>
<dc:creator>Shaw, A. E.</dc:creator>
<dc:creator>Herson, P. S.</dc:creator>
<dc:creator>Bamburg, J. R.</dc:creator>
<dc:creator>Swanson, R. A.</dc:creator>
<dc:date>2018-05-25</dc:date>
<dc:identifier>doi:10.1101/331082</dc:identifier>
<dc:title><![CDATA[Cofilin-actin rod formation in neuronal processes after brain ischemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/331447v1?rss=1">
<title>
<![CDATA[
Early projections of Ebola outbreak size and duration with and without vaccine use in Equateur, Democratic Republic of Congo, as of May 21, 2018 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/331447v1?rss=1</link>
<description><![CDATA[
BackgroundAs of May 27, 2018, 54 cases of Ebola virus disease (EVD) were reported in Equateur Province, Democratic Republic of Congo. We used reported case counts and time series from prior outbreaks to estimate the current outbreak size and duration with and without vaccine use.nnMethodsWe modeled Ebola virus transmission using a stochastic branching process model with a negative binomial distribution, using both estimates of reproduction number R declining from supercritical to subcritical derived from past Ebola outbreaks, as well as a particle filtering method to generate a probabilistic projection of the future course of the outbreak conditioned on its reported trajectory to date; modeled using 0%, 44%, and 62% estimates of vaccination coverage. Additionally, we used the time series for 18 prior Ebola outbreaks from 1976 to 2016 to parameterize a regression model predicting the outbreak size from the number of observed cases from April 4 to May 27.nnResultsWith the stochastic transmission model, we projected a median outbreak size of 78 EVD cases (95% credible interval: 52, 125.4), 86 cases (95% credible interval: 53, 174.3), and 91 cases (95% credible interval: 52, 843.5), using 62%, 44%, and 0% estimates of vaccination coverage. With the regression model, we estimated a median size of 85.0 cases (95% prediction interval: 53.5, 216.6).nnConclusionsThis outbreak has the potential to be the largest outbreak in DRC since 2007. Vaccines are projected to limit outbreak size and duration but are only part of prevention, control, and care strategies.
]]></description>
<dc:creator>Kelly, J. D.</dc:creator>
<dc:creator>Worden, L.</dc:creator>
<dc:creator>Wannier, R.</dc:creator>
<dc:creator>Hoff, N. A.</dc:creator>
<dc:creator>Mukadi, P.</dc:creator>
<dc:creator>Sinai, C.</dc:creator>
<dc:creator>Ackley, S.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Gao, D.</dc:creator>
<dc:creator>Rutherford, G. W.</dc:creator>
<dc:creator>Lietman, T. M.</dc:creator>
<dc:creator>Muyembe-Tamfum, J. J.</dc:creator>
<dc:creator>Rimoin, A. W.</dc:creator>
<dc:creator>Porco, T. C.</dc:creator>
<dc:date>2018-05-26</dc:date>
<dc:identifier>doi:10.1101/331447</dc:identifier>
<dc:title><![CDATA[Early projections of Ebola outbreak size and duration with and without vaccine use in Equateur, Democratic Republic of Congo, as of May 21, 2018]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/332528v1?rss=1">
<title>
<![CDATA[
A genetically-encoded fluorescent sensor enables rapid and specific detection of dopamine in flies, fish, and mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/332528v1?rss=1</link>
<description><![CDATA[
Dopamine (DA) is a central monoamine neurotransmitter involved in many physiological and pathological processes. A longstanding yet largely unmet goal is to measure DA changes reliably and specifically with high spatiotemporal precision, particularly in animals executing complex behaviors. Here we report the development of novel genetically-encoded GPCR-Activation-Based-DA (GRABDA) sensors that enable these measurements. In response to extracellular DA rises, GRABDA sensors exhibit large fluorescence increases ({Delta}F/F0[~]90%) with sub-second kinetics, nanomolar to sub-micromolar affinities, and excellent molecular specificity. Importantly, GRABDA sensors can resolve a single-electrical-stimulus evoked DA release in mouse brain slices, and detect endogenous DA release in the intact brains of flies, fish, and mice. In freely-behaving mice, GRABDA sensors readily report optogenetically-elicited nigrostriatal DA release and depict dynamic mesoaccumbens DA changes during Pavlovian conditioning or during sexual behaviors. Thus, GRABDA sensors enable spatiotemporal precise measurements of DA dynamics in a variety of model organisms while exhibiting complex behaviors.
]]></description>
<dc:creator>Sun, F.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:creator>Jing, M.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Feng, J.</dc:creator>
<dc:creator>Owen, S. F.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Yamaguchi, T.</dc:creator>
<dc:creator>Yong, Z.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Peng, W.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Lin, D.</dc:creator>
<dc:creator>Xu, M.</dc:creator>
<dc:creator>Kreitzer, A. C.</dc:creator>
<dc:creator>Cui, G.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:date>2018-05-31</dc:date>
<dc:identifier>doi:10.1101/332528</dc:identifier>
<dc:title><![CDATA[A genetically-encoded fluorescent sensor enables rapid and specific detection of dopamine in flies, fish, and mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/332767v1?rss=1">
<title>
<![CDATA[
Modeling meiotic homolog pairing: increased fidelity from a tug of war between telomere forces and a pairing-based Brownian ratchet 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/332767v1?rss=1</link>
<description><![CDATA[
Meiotic homolog pairing involves associations between homologous DNA regions scattered along the length of a chromosome. When homologs associate, they tend to do so by a processive zippering processive, which apparently results from avidity effects. Using a computational model, we show that this avidity-driven processive zippering reduces the selectivity of pairing. When active random forces are applied to telomeres, this drop in selectivity is eliminated in a force-dependent manner. Further simulations suggest that active telomere forces are engaged in a tug-of-war against zippering, which can be interpreted as a Brownian ratchet with a stall force that depends on the dissociation constant of pairing. When perfectly homologous regions of high affinity compete with homeologous regions of lower affinity, the affinity difference can be amplified through this tug of war effect provided the telomere force acts in a range that is strong enough to oppose zippering of homeologs while still permitting zippering of correct homologs. The degree of unzippering depends on the radius of the nucleus, such that complete unzippering of homeologous regions can only take place if the nucleus is large enough to pull the two chromosomes completely apart. A picture of meiotic pairing thus emerges that is fundamentally mechanical in nature, possibly explaining the purpose of active telomere forces, increased nuclear diameter, and the presence of "Maverick" chromosomes in meiosis.
]]></description>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:creator>Fung, J. C.</dc:creator>
<dc:date>2018-05-28</dc:date>
<dc:identifier>doi:10.1101/332767</dc:identifier>
<dc:title><![CDATA[Modeling meiotic homolog pairing: increased fidelity from a tug of war between telomere forces and a pairing-based Brownian ratchet]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/333021v1?rss=1">
<title>
<![CDATA[
Rapid Therapeutic Recommendations in the Context of a Global Public Health Crisis using Translational Bioinformatics Approaches: A proof-of-concept study using Nipah Virus Infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/333021v1?rss=1</link>
<description><![CDATA[
We live in a world of emerging new diseases and old diseases resurging in more aggressive forms. Drug development by pharmaceutical companies is a market-driven and costly endeavor, and thus it is often a challenge when drugs are needed for diseases endemic only to certain regions or which affect only a few patients. However, biomedical open data is accessible and reusable for reanalysis and generation of a new hypotheses and discovery. In this study, we leverage biomedical data and tools to analyze available data on Nipah Virus (NiV) infection. NiV infection is an emerging zoonosis that is transmissible to humans and is associated with high mortality rates. In this study, explored the application of computational drug repositioning and chemogenomic enrichment analyses using host transcriptome data to match drugs that could reverse the virus-induced gene signature. We performed analyses using two gene signatures: i) A previously published gene signature (n=34), and ii) a gene signature generated using the characteristic direction method (n= 5,533). Our predictive framework suggests that several drugs including FDA approved therapies like beclometasone, trihexyphenidyl, S-propranolol etc. could modulate the NiV infection induced gene signatures in endothelial cells. A target specific analysis of CXCL10 also suggests the potential application of Eldelumab, an investigative therapy for Crohns disease and ulcerative colitis, as a putative candidate for drug repositioning. To conclude, we also discuss challenges and opportunities in clinical trials (n-of-1 and adaptive trials) for repositioned drugs. Further follow-up studies including biochemical assays and clinical trials are required to identify effective therapies for clinical use. Our proof-of-concept study highlights that translational bioinformatics methods including gene expression analyses and computational drug repositioning could augment epidemiological investigations in the context of an emerging disease with no effective treatment.
]]></description>
<dc:creator>Shameer, K.</dc:creator>
<dc:creator>Johnson, K. W.</dc:creator>
<dc:creator>Readhead, B.</dc:creator>
<dc:creator>Glicksberg, B.</dc:creator>
<dc:creator>McCallum, C.</dc:creator>
<dc:creator>R, A.</dc:creator>
<dc:creator>Hirsch, J.</dc:creator>
<dc:creator>Bock, K.</dc:creator>
<dc:creator>Chelico, J.</dc:creator>
<dc:creator>Hajizadeh, N.</dc:creator>
<dc:creator>Oppenheim, M.</dc:creator>
<dc:creator>Dudley, J.</dc:creator>
<dc:date>2018-05-29</dc:date>
<dc:identifier>doi:10.1101/333021</dc:identifier>
<dc:title><![CDATA[Rapid Therapeutic Recommendations in the Context of a Global Public Health Crisis using Translational Bioinformatics Approaches: A proof-of-concept study using Nipah Virus Infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/334359v1?rss=1">
<title>
<![CDATA[
Loss of the Wnt/β-catenin pathway in microglia of the developing brain drives pro-inflammatory activation leading to white matter injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/334359v1?rss=1</link>
<description><![CDATA[
Microglia of the developing brain have unique functional properties but how their activation states is regulated is poorly understood. Inflammatory activation of microglia in the still-developing brain of preterm born infants is associated with permanent neurological sequelae in 9 million infants every year. Investigating the regulators of microglial activation in the developing brain with multiple models of neuroinflammation-mediated injury and primary human microglia we found that a reduction in Wnt/{beta}-catenin signalling is necessary and sufficient to drive an oligodendrocyte-injurious microglial phenotype. We validated in a cohort of preterm born infants that genomic variation in the WNT pathway is associated with the levels of connectivity found in their brains. Using a Wnt agonist delivered by a BBB penetrant microglia-specific targeting nanocarrier we prevented in our animal model the pro-inflammatory microglial activation, white matter injury and behavioural deficits. Collectively, these data validate that the Wnt pathway regulates microglial activation, is critical in the evolution of an important form of human brain injury and is a viable therapeutic target.
]]></description>
<dc:creator>Van Steenwinckel, J.</dc:creator>
<dc:creator>Schang, A.-L.</dc:creator>
<dc:creator>Krishnan, M. L.</dc:creator>
<dc:creator>Degos, V.</dc:creator>
<dc:creator>Delahaye-Duriez, A.</dc:creator>
<dc:creator>Bokobza, C.</dc:creator>
<dc:creator>Verdonk, F.</dc:creator>
<dc:creator>Montane, A.</dc:creator>
<dc:creator>Sigaut, S.</dc:creator>
<dc:creator>Hennebert, O.</dc:creator>
<dc:creator>Lebon, S.</dc:creator>
<dc:creator>Schwendimann, L.</dc:creator>
<dc:creator>Le Charpentier, T.</dc:creator>
<dc:creator>Hassan-Abdi, R.</dc:creator>
<dc:creator>Ball, G.</dc:creator>
<dc:creator>Aljabar, P.</dc:creator>
<dc:creator>Saxena, A.</dc:creator>
<dc:creator>Holloway, R. K.</dc:creator>
<dc:creator>Birchmeier, W.</dc:creator>
<dc:creator>Miron, V.</dc:creator>
<dc:creator>Rowitch, D.</dc:creator>
<dc:creator>Chretien, F.</dc:creator>
<dc:creator>Leconte, C.</dc:creator>
<dc:creator>Besson, V. C.</dc:creator>
<dc:creator>Petretto, E. G.</dc:creator>
<dc:creator>Edwards, A. D.</dc:creator>
<dc:creator>Hagberg, H.</dc:creator>
<dc:creator>Soussi-Yanicostas, N.</dc:creator>
<dc:creator>Fleiss, B.</dc:creator>
<dc:creator>Gressens, P.</dc:creator>
<dc:date>2018-05-31</dc:date>
<dc:identifier>doi:10.1101/334359</dc:identifier>
<dc:title><![CDATA[Loss of the Wnt/β-catenin pathway in microglia of the developing brain drives pro-inflammatory activation leading to white matter injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/335729v1?rss=1">
<title>
<![CDATA[
CRISPR-Cas9 interrogation of a putative fetal globin repressor in human erythroid cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/335729v1?rss=1</link>
<description><![CDATA[
Sickle Cell Disease and {beta}-thalassemia, which are caused by defective or deficient adult {beta}-globin (HBB) respectively, are the most common serious genetic blood diseases in the world. Expression of the fetal {beta}-like globin, also known as {gamma}-globin, can ameliorate both disorders by serving in place of the adult {beta}-globin. Here we use CRISPR-Cas9 gene editing to explore a putative {gamma}-globin silencer region identified by comparison of naturally-occurring deletion mutations associated with up-regulated {gamma}-globin. We find that deletion of a 1.7 kb consensus element or select 350 bp sub-regions from bulk populations of cells increases levels of fetal hemoglobin (HbF) or {gamma}-globin. Screening of individual sgRNAs in one sub-region revealed three single guides that caused mild increases in {gamma}-globin expression. However, clonal cell lines with the 1.7 kb region deleted did not up-regulate {gamma}-globin and neither did lines with either of two of sub-regions identified in the screen deleted. These data suggest that the region is not an autonomous {gamma}-globin silencer, and thus by itself is not a suitable therapeutic target in the {beta}-hemoglobinopathies.
]]></description>
<dc:creator>Chung, J. E.</dc:creator>
<dc:creator>Magis, W.</dc:creator>
<dc:creator>Vu, J.</dc:creator>
<dc:creator>Heo, S.-J.</dc:creator>
<dc:creator>Wartiovaara, K.</dc:creator>
<dc:creator>Walters, M. C.</dc:creator>
<dc:creator>Kurita, R.</dc:creator>
<dc:creator>Nakamura, Y.</dc:creator>
<dc:creator>Boffelli, D.</dc:creator>
<dc:creator>Martin, D. I.</dc:creator>
<dc:creator>Corn, J. E.</dc:creator>
<dc:creator>Dewitt, M. A.</dc:creator>
<dc:date>2018-05-31</dc:date>
<dc:identifier>doi:10.1101/335729</dc:identifier>
<dc:title><![CDATA[CRISPR-Cas9 interrogation of a putative fetal globin repressor in human erythroid cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/340943v1?rss=1">
<title>
<![CDATA[
Deep learning models reveal internal structure and diverse computations in the retina under natural scenes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/340943v1?rss=1</link>
<description><![CDATA[
Understanding how the visual system encodes natural scenes is a fundamental goal of sensory neuroscience. We show here that a three-layer network model predicts the retinal response to natural scenes with an accuracy nearing the fundamental limits of predictability. The models internal structure is interpretable, in that model units are highly correlated with interneurons recorded separately and not used to fit the model. We further show the ethological relevance to natural visual processing of a diverse set of phenomena of complex motion encoding, adaptation and predictive coding. Our analysis uncovers a fast timescale of visual processing that is inaccessible directly from experimental data, showing unexpectedly that ganglion cells signal in distinct modes by rapidly (< 0.1 s) switching their selectivity for direction of motion, orientation, location and the sign of intensity. A new approach that decomposes ganglion cell responses into the contribution of interneurons reveals how the latent effects of parallel retinal circuits generate the response to any possible stimulus. These results reveal extremely flexible and rapid dynamics of the retinal code for natural visual stimuli, explaining the need for a large set of interneuron pathways to generate the dynamic neural code for natural scenes.
]]></description>
<dc:creator>Maheswaranathan, N.</dc:creator>
<dc:creator>McIntosh, L. T.</dc:creator>
<dc:creator>Kastner, D. B.</dc:creator>
<dc:creator>Melander, J.</dc:creator>
<dc:creator>Brezovec, L.</dc:creator>
<dc:creator>Nayebi, A.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Ganguli, S.</dc:creator>
<dc:creator>Baccus, S. A.</dc:creator>
<dc:date>2018-06-08</dc:date>
<dc:identifier>doi:10.1101/340943</dc:identifier>
<dc:title><![CDATA[Deep learning models reveal internal structure and diverse computations in the retina under natural scenes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/343293v1?rss=1">
<title>
<![CDATA[
New genetic signals for lung function highlight pathways and pleiotropy, and chronic obstructive pulmonary disease associations across multiple ancestries 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/343293v1?rss=1</link>
<description><![CDATA[
Reduced lung function predicts mortality and is key to the diagnosis of COPD. In a genome-wide association study in 400,102 individuals of European ancestry, we define 279 lung function signals, one-half of which are new. In combination these variants strongly predict COPD in deeply-phenotyped patient populations. Furthermore, the combined effect of these variants showed generalisability across smokers and never-smokers, and across ancestral groups. We highlight biological pathways, known and potential drug targets for COPD and, in phenome-wide association studies, autoimmune-related and other pleiotropic effects of lung function associated variants. This new genetic evidence has potential to improve future preventive and therapeutic strategies for COPD.
]]></description>
<dc:creator>Shrine, N.</dc:creator>
<dc:creator>Guyatt, A. L.</dc:creator>
<dc:creator>Erzurumluoglu, A. M.</dc:creator>
<dc:creator>Jackson, V. E.</dc:creator>
<dc:creator>Hobbs, B. D.</dc:creator>
<dc:creator>Melbourne, C.</dc:creator>
<dc:creator>Batini, C.</dc:creator>
<dc:creator>Fawcett, K. A.</dc:creator>
<dc:creator>Song, K.</dc:creator>
<dc:creator>Sakornsakolpat, P.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Boxall, R.</dc:creator>
<dc:creator>Reeve, N. F.</dc:creator>
<dc:creator>Obeidat, M.</dc:creator>
<dc:creator>Zhao, J. H.</dc:creator>
<dc:creator>Wielscher, M.</dc:creator>
<dc:creator>Understanding Society Scientific Group,</dc:creator>
<dc:creator>Weiss, S.</dc:creator>
<dc:creator>Kentistou, K. A.</dc:creator>
<dc:creator>Cook, J. P.</dc:creator>
<dc:creator>Sun, B. B.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Hui, J.</dc:creator>
<dc:creator>Karrasch, S.</dc:creator>
<dc:creator>Imboden, M.</dc:creator>
<dc:creator>Harris, S. E.</dc:creator>
<dc:creator>Marten, J.</dc:creator>
<dc:creator>Enroth, S.</dc:creator>
<dc:creator>Kerr, S. M.</dc:creator>
<dc:creator>Surakka, I.</dc:creator>
<dc:creator>Vitart, V.</dc:creator>
<dc:creator>Lehtimäki, T.</dc:creator>
<dc:creator>Allen, R. J.</dc:creator>
<dc:creator>Bakke, P. S.</dc:creator>
<dc:creator>Beaty, T. H.</dc:creator>
<dc:creator>Bleecker, E. R.</dc:creator>
<dc:creator>Bosse, Y.</dc:creator>
<dc:creator>Brandsma, C.-A.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Crapo, J. D.</dc:creator>
<dc:creator>Danesh, J.</dc:creator>
<dc:creator>DeMeo,</dc:creator>
<dc:date>2018-06-12</dc:date>
<dc:identifier>doi:10.1101/343293</dc:identifier>
<dc:title><![CDATA[New genetic signals for lung function highlight pathways and pleiotropy, and chronic obstructive pulmonary disease associations across multiple ancestries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/345629v1?rss=1">
<title>
<![CDATA[
The Evolution of White Matter Microstructural Changes After Mild Traumatic Brain Injury: A Longitudinal DTI and NODDI Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/345629v1?rss=1</link>
<description><![CDATA[
Neuroimaging biomarkers show promise for improving precision diagnosis and prognosis after mild traumatic brain injury (mTBI), but none has yet been adopted in routine clinical practice. Biophysical modeling of multishell diffusion MRI, using the neurite orientation dispersion and density imaging (NODDI) framework, may improve upon conventional diffusion tensor imaging (DTI) in revealing subtle patterns of underlying white matter microstructural pathology, such as diffuse axonal injury (DAI) and neuroinflammation, that are important for detecting mTBI and determining patient outcome. With a cross-sectional and longitudinal design, we assessed structural MRI, DTI and NODDI in 40 mTBI patients at 2 weeks and 6 months after injury and 14 matched control participants with orthopedic trauma but not suffering from mTBI at 2 weeks. Self-reported and performance-based cognitive measures assessing postconcussive symptoms, memory, executive functions and processing speed were investigated in post-acute and chronic phase after injury for the mTBI subjects. Machine learning analysis was used to identify mTBI patients with the best neuropsychological improvement over time and relate this outcome to DTI and NODDI biomarkers. In the cross-sectional comparison with the trauma control group at 2 weeks post-injury, mTBI patients showed decreased fractional anisotropy (FA) and increased mean diffusivity (MD) on DTI mainly in anterior tracts that corresponded to white matter regions of elevated free water fraction (FISO) on NODDI, signifying vasogenic edema. Patients showed decreases from 2 weeks to 6 months in white matter neurite density on NODDI, predominantly in posterior tracts. No significant longitudinal changes in DTI metrics were observed. The machine learning analysis divided the mTBI patients into two groups based on their recovery. Voxel-wise group comparison revealed associations between white matter orientation dispersion index (ODI) and FISO with degree and trajectory of improvement within the mTBI group. In conclusion, white matter FA and MD alterations early after mTBI might reflect vasogenic edema, as shown by elevated free water on NODDI. Longer-term declines in neurite density on NODDI suggest progressive axonal degeneration due to DAI, especially in tracts known to be integral to the structural connectome. Overall, these results show that the NODDI parameters appear to be more sensitive to longitudinal changes than DTI metrics. Thus, NODDI merits further study in larger cohorts for mTBI diagnosis, prognosis and treatment monitoring.
]]></description>
<dc:creator>Palacios, E.</dc:creator>
<dc:creator>Owen, J. P.</dc:creator>
<dc:creator>Yuh, E. L.</dc:creator>
<dc:creator>Wang, M. B.</dc:creator>
<dc:creator>Vassar, M. J.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Diaz-Arrastia, R.</dc:creator>
<dc:creator>Giacino, J. T.</dc:creator>
<dc:creator>Okonkwo, D. O.</dc:creator>
<dc:creator>Robertson, C. S.</dc:creator>
<dc:creator>Stein, M. B.</dc:creator>
<dc:creator>Temkin, N.</dc:creator>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>McCrea, M.</dc:creator>
<dc:creator>Mac Donald, C. L.</dc:creator>
<dc:creator>Manley, G. T.</dc:creator>
<dc:creator>Mukherjee, P.</dc:creator>
<dc:creator>TRACK-TBI Investigators,</dc:creator>
<dc:date>2018-06-14</dc:date>
<dc:identifier>doi:10.1101/345629</dc:identifier>
<dc:title><![CDATA[The Evolution of White Matter Microstructural Changes After Mild Traumatic Brain Injury: A Longitudinal DTI and NODDI Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/346213v1?rss=1">
<title>
<![CDATA[
Direct evidence for prediction signals in frontal cortex independent of prediction error 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/346213v1?rss=1</link>
<description><![CDATA[
Predictive coding (PC) has been suggested as one of the main mechanisms used by brains to interact with complex environments. PC theories posit top-down prediction signals, which are compared with actual outcomes, yielding in turn prediction-error signals, which are used, bottom-up, to modify the ensuing predictions. However, disentangling prediction from prediction-error signals has been challenging. Critically, while many studies found indirect evidence for predictive coding in the form of prediction-error signals, direct evidence for the prediction signal is mostly lacking. Here we provide clear evidence, obtained from intracranial cortical recordings in human surgical patients, that the human lateral prefrontal cortex generates prediction signals while anticipating an event. Patients listened to task-irrelevant sequences of repetitive tones including infrequent predictable or unpredictable pitch deviants. The amplitude of high frequency broadband (HFB) neural activity was decreased prior to the onset of expected relative to unexpected deviants in the frontal cortex only, and its amplitude was sensitive to the increasing likelihood of deviants following longer trains of standards in the unpredictable condition. Single trial HFB amplitudes predicted deviations and correlated with post-stimulus response to deviations. These results provide direct evidence for frontal cortex prediction signals independent of prediction-error signals.
]]></description>
<dc:creator>Duerschmid, S.</dc:creator>
<dc:creator>Reichert, C.</dc:creator>
<dc:creator>Hinrichs, H.</dc:creator>
<dc:creator>Heinze, H.-J.</dc:creator>
<dc:creator>Kirsch, H. E.</dc:creator>
<dc:creator>Knight, R. T.</dc:creator>
<dc:creator>Deouell, L. Y.</dc:creator>
<dc:date>2018-06-14</dc:date>
<dc:identifier>doi:10.1101/346213</dc:identifier>
<dc:title><![CDATA[Direct evidence for prediction signals in frontal cortex independent of prediction error]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/355628v1?rss=1">
<title>
<![CDATA[
Sensory Over-Responsivity: Parent Report, Direct Assessment Measures, and Neural Architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/355628v1?rss=1</link>
<description><![CDATA[
BackgroundSensory processing differences are common across neurodevelopmental disorders. Thus, reliable measures are needed to understand biologic underpinnings of these differences. This study aims to define a scoring methodology specific to tactile (TOR) and auditory (AOR) over-responsivity. Second, using MRI Diffusion Tensor Imaging, we seek to determine whether children with AOR show measurable differences in their white matter integrity.nnMethodsThis study includes children with AOR and TOR from a mixed neurodevelopmental disorders cohort including autism and sensory processing dysfunction (n= 176) as well as neurotypical children (n= 128). We established cut-off scores for over-responsivity using the parent report: Short Sensory Profile (SSP), and the direct assessment: Sensory Processing-Three Dimensions:Assessment (SP-3D:A). Group comparisons, based on AOR phenotype, were then conducted comparing the white matter fractional anisotropy in 23 regions of interest.nnResultsUsing the direct assessment, 31% of the children with neurodevelopmental disorders had AOR and 27% had TOR. The Inter-test-agreement between SSP and SP-3D:A for AOR was 65% and TOR was 50%. Children with AOR had three white matter tracts showing decreased fractional anisotropy relative to children without AOR.nnConclusionsThis study identified cut scores for AOR and TOR using the SSP parent report and SP-3D:A observation. A combination of questionnaire and direct observation measures should be used in clinical and research settings. The SSP parent report and SP-3D:A direct observation ratings overlapped moderately for sensory related behaviors. Based on these initial structural neuroimaging results, we suggest a putative neural network may contribute to AOR.
]]></description>
<dc:creator>Tavassoli, T.</dc:creator>
<dc:creator>Brandes-Aitken, A.</dc:creator>
<dc:creator>Chu, R.</dc:creator>
<dc:creator>Porter, L.</dc:creator>
<dc:creator>Schoen, S.</dc:creator>
<dc:creator>Miller, L. J.</dc:creator>
<dc:creator>Gerdes, M. R.</dc:creator>
<dc:creator>Owen, J.</dc:creator>
<dc:creator>Mukherjee, P.</dc:creator>
<dc:creator>Marco, E. J.</dc:creator>
<dc:date>2018-06-28</dc:date>
<dc:identifier>doi:10.1101/355628</dc:identifier>
<dc:title><![CDATA[Sensory Over-Responsivity: Parent Report, Direct Assessment Measures, and Neural Architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/361071v1?rss=1">
<title>
<![CDATA[
Feedback-mediated signal conversion promotes viral fitness 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/361071v1?rss=1</link>
<description><![CDATA[
A fundamental signal-processing problem is how biological systems maintain phenotypic states (i.e., canalization) long after degradation of initial catalyst signals. For example, to efficiently replicate, herpesviruses (e.g., human cytomegalovirus, HCMV) rapidly counteract cell-mediated silencing using trans-activators packaged in the tegument of the infecting virion particle. But, the activity of these tegument trans-activators is inherently transient--they undergo immediate proteolysis but delayed synthesis--and how transient activation sustains lytic viral gene expression despite cell-mediated silencing is unclear. By constructing a two-color, conditional-feedback HCMV mutant, we find that positive feedback in HCMVs Immediate Early 1 (IE1) protein is of sufficient strength to sustain HCMV lytic expression. Single-cell time-lapse imaging and mathematical modeling show that IE1 positive feedback converts transient transactivation signals from tegument pp71 proteins into sustained lytic expression, which is obligate for efficient viral replication, whereas attenuating feedback decreases fitness by promoting a reversible silenced state. Together, these results identify a regulatory mechanism enabling herpesviruses to sustain expression despite transient activation signals--akin to early electronic transistors--and expose a potential target for therapeutic intervention.
]]></description>
<dc:creator>Vardi, N.</dc:creator>
<dc:creator>Chaturvedi, S.</dc:creator>
<dc:creator>Weinberger, L. S.</dc:creator>
<dc:date>2018-07-03</dc:date>
<dc:identifier>doi:10.1101/361071</dc:identifier>
<dc:title><![CDATA[Feedback-mediated signal conversion promotes viral fitness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/364315v1?rss=1">
<title>
<![CDATA[
Heterogeneity of transposon expression and activation of the repressive network in human fetal germ cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/364315v1?rss=1</link>
<description><![CDATA[
Epigenetic resetting in germ cells during development leads to the de-repression of transposable elements (TEs). piRNAs protect fetal germ cells from potentially harmful TEs by targeted destruction of mRNA and deposition of repressive epigenetic marks. Here we provide the first evidence for an active piRNA pathway and TE repression in germ cells of human fetal testis. We identify pre-pachytene piRNAs with features of secondary amplification that map most abundantly to L1 family TEs. We find that L1-ORF1p expression is heterogeneous in fetal germ cells, peaks at mid-gestation and declines concomitantly with increasing levels of piRNAs and H3K9me3, as well as nuclear localization of HIWI2. Surprisingly, following this decline, the same cells with accumulation of L1-ORF1p display highest levels of HIWI2 and H3K9me3, whereas L1-ORF1p low cells are also low in HIWI2 and H3K9me3. Conversely, earlier in development, the germ cells lacking L1-ORF1p express high levels of the chaperone HSP90a. We propose that a subset of HSP90a-armed germ cells resists L1 expression, whereas only those vulnerable L1-expressing germ cells activate the PIWI-piRNA repression pathway which leads to epigenetic silencing of L1 via H3K9me3.
]]></description>
<dc:creator>Reznik, B.</dc:creator>
<dc:creator>Cincotta, S. A.</dc:creator>
<dc:creator>Jaszczak, R. G.</dc:creator>
<dc:creator>Mateo, L. J.</dc:creator>
<dc:creator>Shen, J.</dc:creator>
<dc:creator>Cao, M.</dc:creator>
<dc:creator>Bascin, L.</dc:creator>
<dc:creator>Ye, P.</dc:creator>
<dc:creator>An, W.</dc:creator>
<dc:creator>Laird, D.</dc:creator>
<dc:date>2018-07-07</dc:date>
<dc:identifier>doi:10.1101/364315</dc:identifier>
<dc:title><![CDATA[Heterogeneity of transposon expression and activation of the repressive network in human fetal germ cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/366039v1?rss=1">
<title>
<![CDATA[
Smart bone plates can monitor fracture healing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/366039v1?rss=1</link>
<description><![CDATA[
One Sentence SummaryElectrical impedance measurements using microscale sensors implanted in two mouse fracture models tracked longitudinal differences between individual mice with proper healing and mice experiencing poor healing, laying the groundwork for translation to the clinic through integration into fracture fixation implants (i.e. instrumented bone plates).nnAbstractThere are currently no standardized methods for monitoring fracture healing. While histological studies can clearly identify the tissues found in the four stages of repair, in practice surgeons rely on X-ray, which is only useful at later stages of healing after mineralization has occurred. As electrical impedance spectroscopy (EIS) has previously been used to distinguish tissue types during healing, we hypothesized that microscale sensors embedded in the fracture callus could track the changing tissue with high sensitivity. Using in vivo mouse fracture models, we present the first evidence that microscale instrumented implants provide a route for post-operative fracture monitoring. In this study, we implanted sensors in mouse long bone fractures fixed with either external fixators or bone plates. EIS measurements taken across two electrodes implanted in the fracture gap were able to track longitudinal differences between individual mice with proper healing and mice experiencing poor healing. We additionally present an equivalent circuit model that combines the EIS data in order to classify healing states of fractures. Lastly, we show that EIS measures are strongly correlated with standard {micro}CT measures of healing and that these correlations validate clinically-relevant operating frequencies for implementation of this technique. The data from these two models demonstrate that this technique can be translated to the clinic through integration into current fracture management strategies such as bone plating, providing physicians with quantitative information about the state of a fracture to guide clinical decision-making for patients.
]]></description>
<dc:creator>Lin, M. C.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Marmor, M.</dc:creator>
<dc:creator>Herfat, S. T.</dc:creator>
<dc:creator>Bahney, C. S.</dc:creator>
<dc:creator>Maharbiz, M. M.</dc:creator>
<dc:date>2018-07-10</dc:date>
<dc:identifier>doi:10.1101/366039</dc:identifier>
<dc:title><![CDATA[Smart bone plates can monitor fracture healing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/366781v1?rss=1">
<title>
<![CDATA[
The autism-associated gene Scn2a plays an essential role in synaptic stability and learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/366781v1?rss=1</link>
<description><![CDATA[
Autism spectrum disorder (ASD) is strongly associated with de novo gene mutations. One of the most commonly affected genes is SCN2A. ASD-associated SCN2A mutations impair the encoded protein NaV1.2, a sodium channel important for action potential initiation and propagation in developing excitatory cortical neurons. The link between an axonal sodium channel and ASD, a disorder typically attributed to synaptic or transcriptional dysfunction, is unclear. Here, we show NaV1.2 is unexpectedly critical for dendritic excitability and synaptic function in mature pyramidal neurons, in addition to regulating early developmental axonal excitability. NaV1.2 loss reduced action potential backpropagation into dendrites, impairing synaptic plasticity and synaptic stability, even when NaV1.2 expression was disrupted late in development. Furthermore, we identified behavioral impairments in learning and sociability, paralleling observations in children with SCN2A loss. These results reveal a novel dendritic function for NaV1.2, providing insight into cellular mechanisms likely underlying circuit and behavioral dysfunction in ASD.
]]></description>
<dc:creator>Spratt, P.</dc:creator>
<dc:creator>Ben-Shalom, R.</dc:creator>
<dc:creator>Keeshen, C.</dc:creator>
<dc:creator>Burke, K.</dc:creator>
<dc:creator>Clarkson, R.</dc:creator>
<dc:creator>Sanders, S.</dc:creator>
<dc:creator>Bender, K.</dc:creator>
<dc:date>2018-07-10</dc:date>
<dc:identifier>doi:10.1101/366781</dc:identifier>
<dc:title><![CDATA[The autism-associated gene Scn2a plays an essential role in synaptic stability and learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/369322v1?rss=1">
<title>
<![CDATA[
Multi-omics analysis of AML cells treated with azacitidine reveals highly variable cell surface proteome remodeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/369322v1?rss=1</link>
<description><![CDATA[
Myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) are diseases of abnormal hematopoietic differentiation with aberrant epigenetic alterations. Azacitidine (AZA) is a DNA methyltransferase inhibitor (DNMTi) widely used to treat MDS and AML, yet the impact of AZA on the cell surface proteome has not been defined. To identify potential therapeutic targets for use in combination with AZA in AML patients, we investigated the effects of AZA treatment on four AML cell lines (KG1a, HL60, HNT34, and AML193), representing different stages of differentiation. The effect of AZA treatment on these cell lines was characterized at three levels: the DNA methylome (methylation array), the transcriptome (gene expression array), and the cell surface proteome (glycoprotein capture with SILAC labeling). Untreated AML cell lines showed substantial overlap in their methylomes, transcriptomes, and cell surface proteomes. AZA treatment globally reduced DNA methylation in all cell lines, but changes in the transcriptome and surface proteome were subtle and differed among the cell lines. Transcriptome analysis identified five commonly up-regulated coding genes upon AZA treatment in all four cell lines, TRPM4 being the only gene encoding a surface protein, and surface proteomics analysis found no commonly regulated proteins. Gene Set Enrichment Analysis (GSEA) of differentially-regulated RNA and surface proteins showed a decrease in metabolism pathways and an increase in immune defense response pathways. As such, AZA treatment in four AML cell lines had diverse effects at the individual gene and protein level, but converged to regulation of metabolism and immune response at the pathway level. Given the heterogeneous response of AZA in the four cell lines at the gene and protein level, we discuss potential therapeutic strategies for combinations with AZA.
]]></description>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Shi, T.</dc:creator>
<dc:creator>Tang, L.</dc:creator>
<dc:creator>Ni, X.</dc:creator>
<dc:creator>Escoubet, L.</dc:creator>
<dc:creator>MacBeth, K. J.</dc:creator>
<dc:creator>DiMartino, J.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2018-07-14</dc:date>
<dc:identifier>doi:10.1101/369322</dc:identifier>
<dc:title><![CDATA[Multi-omics analysis of AML cells treated with azacitidine reveals highly variable cell surface proteome remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/369587v1?rss=1">
<title>
<![CDATA[
Facile real time detection of membrane colocalization of RAS superfamily GTPase proteins in living cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/369587v1?rss=1</link>
<description><![CDATA[
Members of the RAS family of GTPases (KRAS4A, KRAS4B, HRAS, and NRAS) are the most frequently mutated oncogenes in human cancers. The CAAX motif in the C-terminal hypervariable region (HVR-CAAX domain) contains the cysteine residue that is critical for protein prenylation that enables RAS protein membrane localization, homodimer/oligomer formation, and activation of effector signaling and oncogenic activity. However, it remains unclear if RAS can interact with other prenylated proteins, and if so, how this impacts RAS function. Here we use a newly developed quantifiable Recombinase enhanced Bimolecular Luciferase Complementation strategy (ReBiL2.0) to investigate some of the requirements for RAS superfamily small GTPase protein interactions, and whether this requires cell membrane localization. ReBiL enables such analyses to be done at physiologic expression levels in living cells. Our results confirm that the C-terminal prenylated HVR-CAAX domain is sufficient to direct KRAS and heterologous proteins to colocalize in the cell membrane. We discovered that KRAS also colocalizes with a subset of small GTPase superfamily members including RAC1, RAC2 and DIRAS3 in a prenylation-dependent manner. KRAS colocalization or co-clustering with heterologous proteins can impact KRAS downstream signaling. ReBiL2.0 thus provides a rapid, simple and straightforward method to identify and characterize the colocalization of membrane-associated proteins and to discover agonists and antagonists thereof.
]]></description>
<dc:creator>Li, Y.-C.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Hayes, T. K.</dc:creator>
<dc:creator>Sutton, M. N.</dc:creator>
<dc:creator>Bast, R. C.</dc:creator>
<dc:creator>McCormick, F.</dc:creator>
<dc:creator>Der, C. J.</dc:creator>
<dc:creator>Wahl, G. M.</dc:creator>
<dc:date>2018-07-15</dc:date>
<dc:identifier>doi:10.1101/369587</dc:identifier>
<dc:title><![CDATA[Facile real time detection of membrane colocalization of RAS superfamily GTPase proteins in living cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/370452v1?rss=1">
<title>
<![CDATA[
Massively parallel characterization of regulatory dynamics during neural induction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/370452v1?rss=1</link>
<description><![CDATA[
The molecular components governing neural induction remain largely unknown. Here, we applied a suite of genomic and computational tools to comprehensively identify these components. We performed RNA-seq, ChIP-seq (H3K27ac, H3K27me3) and ATAC-seq on human embryonic stem cells (hESCs) at seven early neural differentiation time points (0-72 hours) and identified thousands of induced genes and regulatory regions. We analyzed the function of ~2,500 selected regions using massively parallel reporter assays at all time points. We found numerous temporal enhancers that correlated with similarly timed epigenetic marks and gene expression. Development of a prioritization method that incorporated all genomic data identified key transcription factors (TFs) involved in neural induction. Individual overexpression of eleven TFs and several combinations in hESCs found novel neural induction regulators. Combined, our results provide a comprehensive map of genes and functional regulatory elements involved in neural induction and identify master regulator TFs that are instrumental for this process.nnOne Sentence SummaryUsing numerous genomic assays and computational tools we characterized the dynamic changes that take place during neural induction.
]]></description>
<dc:creator>Inoue, F.</dc:creator>
<dc:creator>Kreimer, A.</dc:creator>
<dc:creator>Ashuach, T.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Yosef, N.</dc:creator>
<dc:date>2018-07-16</dc:date>
<dc:identifier>doi:10.1101/370452</dc:identifier>
<dc:title><![CDATA[Massively parallel characterization of regulatory dynamics during neural induction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/370791v1?rss=1">
<title>
<![CDATA[
A nucleus-like compartment shields bacteriophage DNA from CRISPR-Cas and restriction nucleases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/370791v1?rss=1</link>
<description><![CDATA[
All viruses require strategies to inhibit or evade the immunity pathways of cells they infect. The viruses that infect bacteria, bacteriophages (phages), must avoid nucleic-acid targeting immune pathways such as CRISPR-Cas and restriction endonucleases to replicate efficiently1. Here, we show that a jumbo phage infecting Pseudomonas aeruginosa, phage {Phi}KZ, is resistant to many immune systems in vivo, including CRISPR-Cas3 (Type I-C), Cas9 (Type II-A), Cas12 (Cpf1, Type V-A), and Type I restriction-modification (R-M) systems. We propose that {Phi}KZ utilizes a nucleus-like shell to protect its DNA from attack. Supporting this, we demonstrate that Cas9 is able to cleave {Phi}KZ DNA in vitro, but not in vivo and that Cas9 is physically occluded from the shell assembled by the phage during infection. Moreover, we demonstrate that the Achilles heel for this phage is the mRNA, as translation occurs outside of the shell, rendering the phage sensitive to the RNA targeting CRISPR-Cas enzyme, Cas13a (C2c2, Type VI-A). Collectively, we propose that the nucleus-like shell assembled by jumbo phages enables potent, broad spectrum evasion of DNA-targeting nucleases.
]]></description>
<dc:creator>Mendoza, S. D.</dc:creator>
<dc:creator>Berry, J. D.</dc:creator>
<dc:creator>Nieweglowska, E. S.</dc:creator>
<dc:creator>Leon, L. M.</dc:creator>
<dc:creator>Agard, D.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2018-07-17</dc:date>
<dc:identifier>doi:10.1101/370791</dc:identifier>
<dc:title><![CDATA[A nucleus-like compartment shields bacteriophage DNA from CRISPR-Cas and restriction nucleases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/373555v1?rss=1">
<title>
<![CDATA[
Narcolepsy risk loci are enriched in immune cells and suggest autoimmune modulation of the T cell receptor repertoire 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/373555v1?rss=1</link>
<description><![CDATA[
Type 1 narcolepsy (T1N) is a neurological condition, in which the death of hypocretin-producing neurons in the lateral hypothalamus leads to excessive daytime sleepiness and symptoms of abnormal Rapid Eye Movement (REM) sleep. Known triggers for narcolepsy are influenza-A infection and associated immunization during the 2009 H1N1 influenza pandemic. Here, we genotyped all remaining consented narcolepsy cases worldwide and assembled this with the existing genotyped individuals. We used this multi-ethnic sample in genome wide association study (GWAS) to dissect disease mechanisms and interactions with environmental triggers (5,339 cases and 20,518 controls). Overall, we found significant associations with HLA (2 GWA significant subloci) and 11 other loci. Six of these other loci have been previously reported (TRA, TRB, CTSH, IFNAR1, ZNF365 and P2RY11) and five are new (PRF1, CD207, SIRPG, IL27 and ZFAND2A). Strikingly, in vaccination-related cases GWA significant effects were found in HLA, TRA, and in a novel variant near SIRPB1. Furthermore, IFNAR1 associated polymorphisms regulated dendritic cell response to influenza-A infection in vitro (p-value =1.92*10-25). A partitioned heritability analysis indicated specific enrichment of functional elements active in cytotoxic and helper T cells. Furthermore, functional analysis showed the genetic variants in TRA and TRB loci act as remarkable strong chain usage QTLs for TRAJ*24 (p-value = 0.0017), TRAJ*28 (p-value = 1.36*10-10) and TRBV*4-2 (p-value = 3.71*10-117). This was further validated in TCR sequencing of 60 narcolepsy cases and 60 DQB1*06:02 positive controls, where chain usage effects were further accentuated. Together these findings show that the autoimmune component in narcolepsy is defined by antigen presentation, mediated through specific T cell receptor chains, and modulated by influenza-A as a critical trigger.
]]></description>
<dc:creator>Ollila, H.</dc:creator>
<dc:creator>Sharon, E.</dc:creator>
<dc:creator>Lin, L.</dc:creator>
<dc:creator>Sinnott-Armstrong, N.</dc:creator>
<dc:creator>Ambati, A.</dc:creator>
<dc:creator>Hillary, R. P.</dc:creator>
<dc:creator>Jolanki, O.</dc:creator>
<dc:creator>Faraco, J.</dc:creator>
<dc:creator>Einen, M.</dc:creator>
<dc:creator>Luo, G.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Han, F.</dc:creator>
<dc:creator>Yan, H.</dc:creator>
<dc:creator>Dong, X. S.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Hong, S.-C.</dc:creator>
<dc:creator>Kim, T. W.</dc:creator>
<dc:creator>Dauvilliers, Y.</dc:creator>
<dc:creator>Barateau, L.</dc:creator>
<dc:creator>Lammers, G. J.</dc:creator>
<dc:creator>Fronczek, R.</dc:creator>
<dc:creator>Mayer, G.</dc:creator>
<dc:creator>Santamaria, J.</dc:creator>
<dc:creator>Arnulf, I.</dc:creator>
<dc:creator>Knudsen, S.</dc:creator>
<dc:creator>Lyamouri-Bredahl, M. K.</dc:creator>
<dc:creator>Thorsby, P. M.</dc:creator>
<dc:creator>Plazzi, G.</dc:creator>
<dc:creator>Pizza, F.</dc:creator>
<dc:creator>Moresco, M.</dc:creator>
<dc:creator>Crowe, C.</dc:creator>
<dc:creator>Van-den-Eeden, S. K.</dc:creator>
<dc:creator>Lecendreux, M.</dc:creator>
<dc:creator>Bourgin, P.</dc:creator>
<dc:creator>Kanbayashi, T.</dc:creator>
<dc:creator>Peraita-Adrados, R.</dc:creator>
<dc:creator>Martinez-Orozco, F. J.</dc:creator>
<dc:creator>Beneto, A.</dc:creator>
<dc:creator>Montplaisir, J.</dc:creator>
<dc:creator>Desautels, A.</dc:creator>
<dc:creator>Huang, Y.-S.</dc:creator>
<dc:creator>Jennum, P.</dc:creator>
<dc:creator>Nevsimalova,</dc:creator>
<dc:date>2018-07-22</dc:date>
<dc:identifier>doi:10.1101/373555</dc:identifier>
<dc:title><![CDATA[Narcolepsy risk loci are enriched in immune cells and suggest autoimmune modulation of the T cell receptor repertoire]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/375360v1?rss=1">
<title>
<![CDATA[
A Metagenomic Comparison of Tracheal Aspirate and Mini-Bronchial Alveolar Lavage for Assessment of Respiratory Microbiota 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/375360v1?rss=1</link>
<description><![CDATA[
Lower respiratory tract infections (LRTI) lead to more deaths each year than any other infectious disease category. Despite this, etiologic LRTI pathogens are infrequently identified due to limitations of existing microbiologic tests. In critically ill patients, non-infectious inflammatory syndromes resembling LRTI further complicate diagnosis. To address the need for improved LRTI diagnostics, we performed metagenomic next-generation sequencing (mNGS) on tracheal aspirates from 92 adults with acute respiratory failure and simultaneously assessed pathogens, the airway microbiome and the host transcriptome. To differentiate pathogens from respiratory commensals, we developed rules-based and logistic regression models (RBM, LRM) in a derivation cohort of 20 patients with LRTI or non-infectious acute respiratory illnesses. When tested in an independent validation cohort of 24 patients, both models achieved accuracies of 95.5%. We next developed pathogen, microbiome diversity, and host gene expression metrics to identify LRTI-positive patients and differentiate them from critically ill controls with non-infectious acute respiratory illnesses. When tested in the validation cohort, the pathogen metric performed with an AUC of 0.96 (95% CI = 0.86 - 1.00), the diversity metric with an AUC of 0.80 (95% CI = 0.63 - 0.98), and the host transcriptional classifier with an AUC of 0.88 (95% CI = 0.75 - 1.00). Combining these achieved a negative predictive value of 100%. This study suggests that a single streamlined protocol offering an integrated genomic portrait of pathogen, microbiome and host transcriptome may hold promise as a novel tool for LRTI diagnosis.nnSIGNIFICANCE STATEMENTLower respiratory tract infections (LRTI) are the leading cause of infectious disease-related death worldwide yet remain challenging to diagnose because of limitations in existing microbiologic tests. In critically ill patients, non-infectious respiratory syndromes that resemble LRTI further complicate diagnosis and confound targeted treatment. To address this, we developed a novel metagenomic sequencing-based approach that simultaneously interrogates three core elements of acute airway infections: the pathogen, airway microbiome and host response. We studied this approach in a prospective cohort of critically ill patients with acute respiratory failure and found that combining pathogen, microbiome and host gene expression metrics achieved accurate LRTI diagnosis and identified etiologic pathogens in patients with clinically identified infections but otherwise negative testing.
]]></description>
<dc:creator>Kalantar, K.</dc:creator>
<dc:creator>Moazed, F.</dc:creator>
<dc:creator>Christenson, S.</dc:creator>
<dc:creator>Wilson, J.</dc:creator>
<dc:creator>Deiss, T.</dc:creator>
<dc:creator>Belzer, A.</dc:creator>
<dc:creator>Vessell, K.</dc:creator>
<dc:creator>Jauregui, A.</dc:creator>
<dc:creator>Boulourchi, S.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Calfee, C.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:date>2018-07-24</dc:date>
<dc:identifier>doi:10.1101/375360</dc:identifier>
<dc:title><![CDATA[A Metagenomic Comparison of Tracheal Aspirate and Mini-Bronchial Alveolar Lavage for Assessment of Respiratory Microbiota]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/383844v1?rss=1">
<title>
<![CDATA[
Lipid associated polygenic enrichment in Alzheimer’s disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/383844v1?rss=1</link>
<description><![CDATA[
Cardiovascular (CV) and lifestyle associated risk factors (RFs) are increasingly recognized as important for Alzheimers disease (AD) pathogenesis. Beyond the [isin]4 allele of apolipoprotein E (APOE), comparatively little is known about whether CV associated genes also increase risk for AD (genetic pleiotropy). Using large genome-wide association studies (GWASs) (total n > 500,000 cases and controls) and validated tools to quantify genetic pleiotropy, we systematically identified single nucleotide polymorphisms (SNPs) jointly associated with AD and one or more CV RFs, namely body mass index (BMI), type 2 diabetes (T2D), coronary artery disease (CAD), waist hip ratio (WHR), total cholesterol (TC), low-density (LDL) and high-density lipoprotein (HDL). In fold enrichment plots, we observed robust genetic enrichment in AD as a function of plasma lipids (TC, LDL, and HDL); we found minimal AD genetic enrichment conditional on BMI, T2D, CAD, and WHR. Beyond APOE, at conjunction FDR < 0.05 we identified 57 SNPs on 19 different chromosomes that were jointly associated with AD and CV outcomes including APOA4, ABCA1, ABCG5, LIPG, and MTCH2/SPI1. We found that common genetic variants influencing AD are associated with multiple CV RFs, at times with a different directionality of effect. Expression of these AD/CV pleiotropic genes was enriched for lipid metabolism processes, over-represented within astrocytes and vascular structures, highly co-expressed, and differentially altered within AD brains. Beyond APOE, we show that the polygenic component of AD is enriched for lipid associated RFs. Rather than a single causal link between genetic loci, RF and the outcome, we found that common genetic variants influencing AD are associated with multiple CV RFs. Our collective findings suggest that a network of genes involved in lipid biology also influence Alzheimers risk.
]]></description>
<dc:creator>Broce, I.</dc:creator>
<dc:creator>Tan, C. H.</dc:creator>
<dc:creator>Fan, C. C.</dc:creator>
<dc:creator>Witoelar, A.</dc:creator>
<dc:creator>Wen, N.</dc:creator>
<dc:creator>Jansen, I.</dc:creator>
<dc:creator>Hess, C.</dc:creator>
<dc:creator>Dillon, W.</dc:creator>
<dc:creator>Glastonbury, C.</dc:creator>
<dc:creator>Glymour, M.</dc:creator>
<dc:creator>Yokoyama, J.</dc:creator>
<dc:creator>Elahi, F.</dc:creator>
<dc:creator>Rabinovici, G.</dc:creator>
<dc:creator>Miller, B.</dc:creator>
<dc:creator>Mormino, E.</dc:creator>
<dc:creator>Sperling, R.</dc:creator>
<dc:creator>Bennett, D.</dc:creator>
<dc:creator>McEvoy, L.</dc:creator>
<dc:creator>Brewer, J.</dc:creator>
<dc:creator>Feldman, H.</dc:creator>
<dc:creator>Posthuma, D.</dc:creator>
<dc:creator>Hyman, B.</dc:creator>
<dc:creator>Schellenberg, G.</dc:creator>
<dc:creator>Yaffe, K.</dc:creator>
<dc:creator>Andreassen, O.</dc:creator>
<dc:creator>Dale, A.</dc:creator>
<dc:creator>Sugrue, L.</dc:creator>
<dc:creator>Karch, C.</dc:creator>
<dc:creator>Desikan, R.</dc:creator>
<dc:date>2018-08-03</dc:date>
<dc:identifier>doi:10.1101/383844</dc:identifier>
<dc:title><![CDATA[Lipid associated polygenic enrichment in Alzheimer’s disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/394056v1?rss=1">
<title>
<![CDATA[
Minor isozymes tailor yeast metabolism to carbon availability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/394056v1?rss=1</link>
<description><![CDATA[
Isozymes are enzymes that differ in sequence but catalyze the same chemical reactions. Despite their apparent redundancy, isozymes are often retained over evolutionary time for reasons that can be unclear. We find that, in yeast, isozymes are strongly enriched in central carbon metabolism. Using a gene expression compendium, we find that many isozyme pairs show anticorrelated expression during the respirofermentative shift, suggesting roles in adapting to changing carbon availability. Building on this observation, we assign function to two minor central carbon isozymes, aconitase 2 (ACO2) and pyruvate kinase 2 (PYK2). ACO2 is expressed during fermentation and proves advantageous when glucose is limiting. PYK2 is expressed during respiration and proves advantageous for growth on three-carbon substrates. PYK2s deletion is rescued by expressing the major pyruvate kinase, but only if that enzyme carries mutations mirroring PYK2s allosteric regulation. Thus, central carbon isozymes enable more precise tailoring of metabolism to changing nutrient availability.nnImportanceGene duplication is one of the main evolutionary drivers of new protein function. However, some gene duplicates have nevertheless persisted long-term without apparent divergence in biochemical function. Further, under standard lab conditions, many isozymes have subtle or no knockout phenotypes. These factors make it hard to assess the unique contributions of individual isozymes to fitness. We therefore developed a method to identify experimental perturbations that could expose such contributions, and applied it to yeast gene expression data, revealing a potential role for a set of yeast isozymes in adapting to changing carbon sources. Our experimental confirmation of distinct roles for two "minor" yeast isozymes, including one with no previously described knockout phenotype, highlight that even apparently redundant paralogs can have important and unique functions, with implications for genome-scale metabolic modeling and systems-level studies of quantitative genetics.
]]></description>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:creator>Gibney, P. A.</dc:creator>
<dc:creator>Botstein, D.</dc:creator>
<dc:creator>Troyanskaya, O.</dc:creator>
<dc:creator>Rabinowitz, J.</dc:creator>
<dc:date>2018-08-17</dc:date>
<dc:identifier>doi:10.1101/394056</dc:identifier>
<dc:title><![CDATA[Minor isozymes tailor yeast metabolism to carbon availability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/394965v1?rss=1">
<title>
<![CDATA[
Challenge data set for macromolecular multi-microcrystallography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/394965v1?rss=1</link>
<description><![CDATA[
A synthetic data set demonstrating a particularly challenging case of indexing ambiguity in the context of radiation damage was generated in order to serve as a standard benchmark and reference point for the ongoing development of new methods and new approaches to solving this problem. Of the 100 short wedges of data only the first 71 are currently necessary to solve the structure by "cheating", or using the correct reference structure as a guide. The total wall-clock time and number of wedges required to solve the structure without cheating is proposed as a metric for the efficacy and efficiency of a given multi-crystal automation pipeline.nnSynopsisA synthetic dataset demonstrating the challenges of combining multiple data sets with indexing ambiguity in the context of heavy radiation damage in multi-crystal macromolecular crystallography was generated and described, and the problems encountered using contemporary data processing programs were summarized.
]]></description>
<dc:creator>Holton, J. M.</dc:creator>
<dc:date>2018-08-18</dc:date>
<dc:identifier>doi:10.1101/394965</dc:identifier>
<dc:title><![CDATA[Challenge data set for macromolecular multi-microcrystallography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/397638v1?rss=1">
<title>
<![CDATA[
GxEMM: Extending linear mixed models to general gene-environment interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/397638v1?rss=1</link>
<description><![CDATA[
Gene-environment interaction (GxE) is a well-known source of non-additive inheritance. GxE can be important in applications ranging from basic functional genomics to precision medical treatment. Further, GxE effects elude inherently-linear LMMs and may explain missing heritability. We propose a simple, unifying mixed model for polygenic interactions (GxEMM) to capture the aggregate effect of small GxE effects spread across the genome. GxEMM extends existing LMMs for GxE in two important ways. First, it extends to arbitrary environmental variables, not just categorical groups. Second, GxEMM can estimate and test for environment-specific heritability. In simulations where the assumptions of existing methods do not hold, we show that GxEMM improves estimates of ordinary and GxE heritability and increases power to test for polygenic GxE. We then use GxEMM to prove that the heritability of major depression (MD) is reduced by stress, which we previously conjectured but could not prove with prior methods, and that a tail of polygenic GxE effects remains unexplained by MD GWAS.
]]></description>
<dc:creator>Dahl, A.</dc:creator>
<dc:creator>Cai, N.</dc:creator>
<dc:creator>Flint, J.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:date>2018-08-22</dc:date>
<dc:identifier>doi:10.1101/397638</dc:identifier>
<dc:title><![CDATA[GxEMM: Extending linear mixed models to general gene-environment interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/398271v1?rss=1">
<title>
<![CDATA[
Multiple Exposures, Reinfection, and Risk of Progression to Active Tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/398271v1?rss=1</link>
<description><![CDATA[
A recent study reported on a tuberculosis outbreak in a largely Inuit village. Among recently infected individuals, exposure to additional active cases was associated with an increasing probability of developing active disease within a year. Using binomial risk models, we evaluated two potential mechanisms by which multiple infections during the first year following initial infection could account for increasing disease risk with increasing exposures. In the reinfection model, multiple exposures have an independent risk of becoming an infection, and infections contribute independently to active disease. In the threshold model, disease risk follows a sigmoidal function with small numbers of exposures conferring a low risk of active disease and large numbers of exposures conferring a high risk. To determine the dynamic impact of reinfection during the early phase of infection, we performed simulations from a modified Reed-Frost model of TB dynamics following spread from an initial number of cases. We parameterized this model with the maximum likelihood estimates from the reinfection and threshold models in addition to the observed distribution of exposures among recent infections. We find that both models can plausibly account for the observed increase in disease risk with increasing exposures, but the threshold model confers a better fit than a nested model without a threshold (p=0.04). Our simulations indicate that multiple exposures during this critical time period can lead to dramatic increases in outbreak size. In order to decrease TB burden in high-prevalence settings, it may be necessary to implement measures aimed at preventing repeated exposures, in addition to preventing primary infection.
]]></description>
<dc:creator>Ackley, S.</dc:creator>
<dc:creator>Lee, R. S.</dc:creator>
<dc:creator>Worden, L.</dc:creator>
<dc:creator>Zwick, E.</dc:creator>
<dc:creator>Porco, T. C.</dc:creator>
<dc:creator>Behr, M. A.</dc:creator>
<dc:creator>Pepperell, C. S.</dc:creator>
<dc:date>2018-08-24</dc:date>
<dc:identifier>doi:10.1101/398271</dc:identifier>
<dc:title><![CDATA[Multiple Exposures, Reinfection, and Risk of Progression to Active Tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/398552v1?rss=1">
<title>
<![CDATA[
Small molecule inhibition of IRE1α kinase/RNase has anti-fibrotic effects in the lung 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/398552v1?rss=1</link>
<description><![CDATA[
Endoplasmic reticulum stress (ER stress) has been implicated in the pathogenesis of idiopathic pulmonary fibrosis (IPF), a disease of progressive fibrosis and respiratory failure. ER stress activates a signaling pathway called the unfolded protein response (UPR) that either restores homeostasis or promotes apoptosis. The bifunctional kinase/RNase IRE1 is a UPR sensor that promotes apoptosis if ER stress remains high (i.e., a "terminal" UPR). Using multiple small molecule inhibitors against IRE1, we show that ER stress-induced apoptosis of murine alveolar epithelial cells can be mitigated in vitro. In vivo, we show that bleomycin exposure to murine lungs causes early ER stress to activate IRE1 and the terminal UPR prior to development of pulmonary fibrosis. Small-molecule IRE1 kinase-inhibiting RNase attenuators (KIRAs) that we developed were used to evaluate the importance of IRE1 activation in bleomycin-induced pulmonary fibrosis. One such KIRA--KIRA7--provided systemically to mice at the time of bleomycin exposure decreases terminal UPR signaling and prevents lung fibrosis. Administration of KIRA7 14 days after bleomycin exposure even promoted the reversal of established fibrosis. Finally, we show that KIRA8, a nanomolar-potent, monoselective KIRA compound derived from a completely different scaffold than KIRA7, likewise promoted reversal of established fibrosis. These results demonstrate that IRE1 may be a promising target in pulmonary fibrosis and that kinase inhibitors of IRE1 may eventually be developed into efficacious anti-fibrotic drugs.
]]></description>
<dc:creator>Thamsen, M.</dc:creator>
<dc:creator>Ghosh, R.</dc:creator>
<dc:creator>Auyeung, V. C.</dc:creator>
<dc:creator>Brumwell, A.</dc:creator>
<dc:creator>Chapman, H. A.</dc:creator>
<dc:creator>Backes, B. J.</dc:creator>
<dc:creator>Perara, G.</dc:creator>
<dc:creator>Maly, D. J.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:creator>Papa, F. R.</dc:creator>
<dc:date>2018-08-23</dc:date>
<dc:identifier>doi:10.1101/398552</dc:identifier>
<dc:title><![CDATA[Small molecule inhibition of IRE1α kinase/RNase has anti-fibrotic effects in the lung]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/399329v1?rss=1">
<title>
<![CDATA[
An open-source control system for in vivo fluorescence measurements from deep-brain structures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/399329v1?rss=1</link>
<description><![CDATA[
BackgroundIntracranial photometry through chronically implanted optical fibers is a widely adopted technique for measuring signals from fluorescent probes in deep-brain structures. The recent proliferation of bright, photo-stable, and specific genetically-encoded fluorescent reporters for calcium and for other neuromodulators has greatly increased the utility and popularity of this technique.nnNew MethodHere we describe an open-source, cost-effective, microcontroller-based solution for controlling optical components in an intracranial photometry system and processing the resulting signal.nnResultsWe show proof-of-principle that this system supports high quality intracranial photometry recordings from dorsal striatum in freely moving mice. A single system supports simultaneous fluorescence measurements in two independent color channels, but multiple systems can be integrated together if additional fluorescence channels are required. This system is designed to work in combination with either commercially available or custom-built optical components. Parts can be purchased for less than one tenth the cost of commercially available alternatives and complete assembly takes less than one day for an inexperienced user.nnComparison with Existing Method(s)Currently available hardware draws on a variety of commercial, custom-built, or hybrid elements for both optical and electronic components. Many of these hardware systems are either specialized and inflexible, or over-engineered and expensive.nnConclusionsThis open-source system increases experimental flexibility while reducing cost relative to current commercially available components. All software and firmware are open-source and customizable, affording a degree of experimental flexibility that is not available in current commercial systems.
]]></description>
<dc:creator>Owen, S. F.</dc:creator>
<dc:creator>Kreitzer, A. C.</dc:creator>
<dc:date>2018-08-23</dc:date>
<dc:identifier>doi:10.1101/399329</dc:identifier>
<dc:title><![CDATA[An open-source control system for in vivo fluorescence measurements from deep-brain structures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/401489v1?rss=1">
<title>
<![CDATA[
Depletion of microbiome-derived molecules in the host using Clostridium genetics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/401489v1?rss=1</link>
<description><![CDATA[
The gut microbiota produce hundreds of molecules that are present at high concentrations in circulation and whose levels vary widely among humans. In most cases, molecule production has not been linked to specific bacterial strains or metabolic pathways, and unraveling the contribution of each molecule to host biology remains difficult. A general system to  toggle molecules in this pool on/off in the host would enable interrogation of the mechanisms by which they modulate host biology and disease processes. Such a system has been elusive due to limitations in the genetic manipulability of Clostridium and its relatives, the source of many molecules in this pool. Here, we describe a method for reliably constructing clean deletions in a model commensal Clostridium, C. sporogenes (Cs), including multiply mutated strains. We demonstrate the utility of this method by using it to  toggle off the production of ten Cs-derived molecules that accumulate in host tissues. By comparing mice colonized by wild-type Cs versus a mutant deficient in the production of branched short-chain fatty acids, we discover a previously unknown IgA-modulatory activity of these abundant microbiome-derived molecules. Our method opens the door to interrogating and sculpting a highly concentrated pool of chemicals from the microbiome.
]]></description>
<dc:creator>Guo, C.-J.</dc:creator>
<dc:creator>Allen, B.</dc:creator>
<dc:creator>Hiam, K.</dc:creator>
<dc:creator>Dodd, D.</dc:creator>
<dc:creator>van Treuren, W.</dc:creator>
<dc:creator>Higginbottom, S.</dc:creator>
<dc:creator>Fischer, C.</dc:creator>
<dc:creator>Sonnenburg, J.</dc:creator>
<dc:creator>Spitzer, M.</dc:creator>
<dc:creator>Fischbach, M.</dc:creator>
<dc:date>2018-08-27</dc:date>
<dc:identifier>doi:10.1101/401489</dc:identifier>
<dc:title><![CDATA[Depletion of microbiome-derived molecules in the host using Clostridium genetics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/403873v1?rss=1">
<title>
<![CDATA[
The 22q11 low copy repeats are characterized by unprecedented size and structure variability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/403873v1?rss=1</link>
<description><![CDATA[
AbstractLow copy repeats (LCRs) are recognized as a significant source of genomic instability, driving genome variability and evolution. The chromosome 22 LCRs (LCR22s) are amongst the most complex regions in the genome and their structure remains unresolved. These LCR22s mediate non-allelic homologous recombination (NAHR) leading to the 22q11 deletion syndrome (22q11DS), causing the most frequent genomic disorder. Using fiber FISH optical mapping, we have de novo assembled the LCR22s in 33 cell lines. We observed a high level of variation in LCR22 structures, including 26 different haplotypes of LCR22A with alleles ranging from 250 Kb to over 2,000 Kb. An additional four haplotypes were detected using Bionano mapping. Further, Bionano maps generated from 154 individuals from different populations suggested significantly different LCR22 haplotype frequencies between populations. Furthermore, haplotype analysis in nine 22q11DS patients resulted in the localization of the NAHR site to a 160 Kb paralog between LCR22A and -D in seven patients and to a 31 Kb region in two individuals with a rearrangement between LCR22A and -B.. This 31 Kb region contains a palindromic AT-rich repeat known to be a driver of chromosomal rearrangements. Our study highlights an unprecedented level of polymorphism in the structure of LCR22s, which are likely still evolving. We present the most comprehensive map of LCR22 variation to date, paving the way towards investigating the role of LCR variation as a driver of 22q11 rearrangements and the phenotypic variability in 22q11DS patients as well as in the general population.
]]></description>
<dc:creator>Demaerel, W.</dc:creator>
<dc:creator>Mostovoy, Y.</dc:creator>
<dc:creator>Yilmaz, F.</dc:creator>
<dc:creator>Vervoort, L.</dc:creator>
<dc:creator>Pastor, S.</dc:creator>
<dc:creator>Hestand, M. S.</dc:creator>
<dc:creator>Swillen, A.</dc:creator>
<dc:creator>Vergaelen, E.</dc:creator>
<dc:creator>Geiger, E. A.</dc:creator>
<dc:creator>Coughlin, C. R.</dc:creator>
<dc:creator>Chow, S. K.</dc:creator>
<dc:creator>McDonald-McGinn, D.</dc:creator>
<dc:creator>Morrow, B.</dc:creator>
<dc:creator>Kwok, P.-Y.</dc:creator>
<dc:creator>Xiao, M.</dc:creator>
<dc:creator>Emanuel, B. S.</dc:creator>
<dc:creator>Shaikh, T. H.</dc:creator>
<dc:creator>Vermeesch, J. R.</dc:creator>
<dc:date>2018-09-12</dc:date>
<dc:identifier>doi:10.1101/403873</dc:identifier>
<dc:title><![CDATA[The 22q11 low copy repeats are characterized by unprecedented size and structure variability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/404350v1?rss=1">
<title>
<![CDATA[
Peripartum cardiomyopathy and hypertensive disorders of pregnancy and cardiovascular events among 1.6 million California pregnancies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/404350v1?rss=1</link>
<description><![CDATA[
BackgroundCardiovascular complications during and soon after pregnancy present an opportunity to assess risk for subsequent cardiovascular disease. We sought to determine whether peripartum cardiomyopathy and hypertensive disorder of pregnancy subtypes predict future myocardial infarction, heart failure, or stroke independent of one another and independent of other risks like gestational diabetes, preterm birth, and intrauterine growth restriction.nnMethods and ResultsThe California Healthcare Cost and Utilization Project database was used to identify all hospitalized pregnancies from 2005-2009, with follow-up through 2011, for a retrospective cohort study. Pregnancies, exposures, covariates and outcomes were defined by ICD-9 codes. Among 1.6 million pregnancies (mean age 28y; median follow-up time to event 2.7y), 558 cases of peripartum cardiomyopathy, 123,603 cases of hypertensive disorders of pregnancy, 107,636 cases of gestational diabetes, 116,768 preterm births, and 23,504 cases of intrauterine growth restriction were observed. Using multivariable Cox proportional hazards models, peripartum cardiomyopathy was independently associated with a 13.0-fold increase in myocardial infarction [95%CI, 4.1-40.9], a 39.2-fold increase in heart failure [95%CI, 30.0-51.9], and a 7.7-fold increase in stroke [95%CI, 2.4-24.0]. Hypertensive disorders of pregnancy were associated with a 1.4 [95%CI, 1.0-2.0] to 7.6 [95%CI, 5.4-10.7] fold higher risk of myocardial infarction, heart failure, and stroke. Gestational diabetes, preterm birth, and intrauterine growth restriction had more modest associations with CVD.nnConclusionsThese findings support close monitoring of women with cardiovascular pregnancy complications for prevention of early subsequent cardiovascular events and further study of mechanisms underlying their development.
]]></description>
<dc:creator>Arnaout, R.</dc:creator>
<dc:creator>Nah, G.</dc:creator>
<dc:creator>Marcus, G.</dc:creator>
<dc:creator>Tseng, Z. H.</dc:creator>
<dc:creator>Foster, E.</dc:creator>
<dc:creator>Harris, I.</dc:creator>
<dc:creator>Dianji, P.</dc:creator>
<dc:creator>Klein, L.</dc:creator>
<dc:creator>Gonzalez, J. M.</dc:creator>
<dc:creator>Parikh, N. I.</dc:creator>
<dc:date>2018-08-30</dc:date>
<dc:identifier>doi:10.1101/404350</dc:identifier>
<dc:title><![CDATA[Peripartum cardiomyopathy and hypertensive disorders of pregnancy and cardiovascular events among 1.6 million California pregnancies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/406132v1?rss=1">
<title>
<![CDATA[
Gelbox - An Interactive Simulation Tool for Gel Electrophoresis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/406132v1?rss=1</link>
<description><![CDATA[
Gel electrophoresis enables separation and visualization of biomolecules such as DNA, RNA, or proteins. Like many powerful tools, mastering the use of gels and making the information gained from them productive, can be dificult. Gelbox is a simulation tool that helps users understand how changing experimental input parameters can affect the data output from gel electrophoresis. Our simulation model handles a wide range of settings, including many "suboptimal" values that may be useful for troubleshooting common mistakes made by novices.
]]></description>
<dc:creator>Gingold, C.</dc:creator>
<dc:creator>Douglas, S. M.</dc:creator>
<dc:date>2018-09-05</dc:date>
<dc:identifier>doi:10.1101/406132</dc:identifier>
<dc:title><![CDATA[Gelbox - An Interactive Simulation Tool for Gel Electrophoresis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/407841v1?rss=1">
<title>
<![CDATA[
Chromatin rewiring mediates programmed evolvability via aneuploidy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/407841v1?rss=1</link>
<description><![CDATA[
Eukaryotes have evolved elaborate mechanisms to ensure that chromosomes segregate with high fidelity during mitosis and meiosis1, and yet specific aneuploidies can be adaptive during environmental stress2,3. Here, we identify a chromatin-based system for inducible aneuploidy in a human pathogen. Candida albicans utilizes chromosome missegregation to acquire resistance to antifungal drugs4,5 and for ploidy reduction after mating6. We discovered that the ancestor of C. albicans and two related pathogens evolved a variant of histone H2A that lacks the conserved phosphorylation site for Bub1 kinase7, a key regulator of chromosome segregation1. Expression of this variant controls the rates of aneuploidy and antibiotic resistance in this species. Moreover, CENP-A/Cse4, the histone H3 that specifies centromeres, is depleted from tetraploid mating products and virtually eliminated from cells exposed to aneuploidy-promoting cues. Thus, changes in chromatin regulation can confer the capacity for rapid evolution in eukaryotes.
]]></description>
<dc:creator>Brimmacombe, C.</dc:creator>
<dc:creator>Burke, J.</dc:creator>
<dc:creator>Parsa, J.-Y.</dc:creator>
<dc:creator>Witchley, J.</dc:creator>
<dc:creator>Burrack, L.</dc:creator>
<dc:creator>Madhani, H.</dc:creator>
<dc:creator>Noble, S.</dc:creator>
<dc:date>2018-09-03</dc:date>
<dc:identifier>doi:10.1101/407841</dc:identifier>
<dc:title><![CDATA[Chromatin rewiring mediates programmed evolvability via aneuploidy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/409722v1?rss=1">
<title>
<![CDATA[
Landscape of stimulation-responsive chromatin across diverse human immune cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/409722v1?rss=1</link>
<description><![CDATA[
The immune system is controlled by a balanced interplay among specialized cell types transitioning between resting and stimulated states. Despite its importance, the regulatory landscape of this system has not yet been fully characterized. To address this gap, we collected ATAC-seq and RNA-seq data under resting and stimulated conditions for 25 immune cell types from peripheral blood of four healthy individuals, and seven cell types from three fetal thymus samples. We found that stimulation caused widespread chromatin remodeling, including a large class of response elements shared between stimulated B and T cells. Furthermore, several autoimmune traits showed significant heritability in stimulation-responsive elements from distinct cell types, highlighting the critical importance of these cell states in autoimmunity. Use of allele-specific read-mapping identified thousands of variants that alter chromatin accessibility in particular conditions. Notably, variants associated with changes in stimulation-specific chromatin accessibility were not enriched for associations with gene expression regulation in whole blood - a tissue commonly used in eQTL studies. Thus, large-scale maps of variants associated with gene regulation lack a condition important for understanding autoimmunity. As a proof-of-principle we identified variant rs6927172, which links stimulated T cell-specific chromatin dysregulation in the TNFAIP3 locus to ulcerative colitis and rheumatoid arthritis. Overall, our results provide a broad resource of chromatin landscape dynamics and highlight the need for large-scale characterization of effects of genetic variation in stimulated cells.
]]></description>
<dc:creator>Calderon, D.</dc:creator>
<dc:creator>Nguyen, M. L. T.</dc:creator>
<dc:creator>Mezger, A.</dc:creator>
<dc:creator>Kathiria, A.</dc:creator>
<dc:creator>Nguyen, V.</dc:creator>
<dc:creator>Lescano, N.</dc:creator>
<dc:creator>Wu, B.</dc:creator>
<dc:creator>Trombetta, J.</dc:creator>
<dc:creator>Ribado, J. V.</dc:creator>
<dc:creator>Knowles, D. A.</dc:creator>
<dc:creator>Gao, Z.</dc:creator>
<dc:creator>Parent, A. V.</dc:creator>
<dc:creator>Burt, T. D.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>Criswell, L. A.</dc:creator>
<dc:creator>Greenleaf, W. J.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:date>2018-09-05</dc:date>
<dc:identifier>doi:10.1101/409722</dc:identifier>
<dc:title><![CDATA[Landscape of stimulation-responsive chromatin across diverse human immune cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/414979v1?rss=1">
<title>
<![CDATA[
Pediatric Brainstem Encephalitis Outbreak Investigation with Metagenomic Next-Generation Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/414979v1?rss=1</link>
<description><![CDATA[
In 2016, Catalonia experienced a pediatric brainstem encephalitis outbreak caused by enterovirus A71 (EV-A71). Conventional testing identified EV in peripheral body sites, but EV was rarely identified in cerebrospinal fluid (CSF). RNA was extracted from CSF (n=20), plasma (n=9), stool (n=15) and nasopharyngeal samples (n=16) from 10 children with brainstem encephalitis or encephalomyelitis and 10 contemporaneous pediatric controls with presumed viral meningitis or encephalitis. Unbiased complementary DNA libraries were sequenced, and microbial pathogens were identified using a custom bioinformatics pipeline. Full-length virus genomes were assembled for phylogenetic analyses. Metagenomic next-generation sequencing (mNGS) was concordant with qRT-PCR for all samples positive by PCR (n=25). In virus-negative samples (n=35), mNGS detected virus in 28.6% (n=10), including 5 CSF samples. mNGS co-detected EV-A71 and another EV in 5 patients. Overall, mNGS increased the proportion of EV-positive samples from 42% (25/60) to 57% (34/60) (McNemars test; p-value = 0.0077). For CSF, mNGS doubled the number of pathogen-positive samples (McNemars test; p-value = 0.074). Using phylogenetic analysis, the outbreak EV-A71 clustered with a neuroinvasive German EV-A71 isolate. Brainstem encephalitis specific, non-synonymous EV-A71 single nucleotide variants were not identified. mNGS demonstrated 100% concordance with clinical qRT-PCR of EV-related brainstem encephalitis and significantly increased the detection of enteroviruses. Our findings increase the probability that neurologic complications observed were virus-induced rather than para-infectious. A comprehensive genomic analysis confirmed that the EV-A71 outbreak strain was closely related to a neuroinvasive German EV-A71 isolate. There were no clear-cut viral genomic differences that discriminated between patients with differing neurologic phenotypes.
]]></description>
<dc:creator>Leon, K. E.</dc:creator>
<dc:creator>Casas-Alba, D.</dc:creator>
<dc:creator>Ramesh, A.</dc:creator>
<dc:creator>Khan, L. M.</dc:creator>
<dc:creator>Launes, C.</dc:creator>
<dc:creator>Sample, H. A.</dc:creator>
<dc:creator>Zorn, K. C.</dc:creator>
<dc:creator>Vallero-Rello, A.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>Munoz-Almagro, C.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:date>2018-09-12</dc:date>
<dc:identifier>doi:10.1101/414979</dc:identifier>
<dc:title><![CDATA[Pediatric Brainstem Encephalitis Outbreak Investigation with Metagenomic Next-Generation Sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/422675v1?rss=1">
<title>
<![CDATA[
Detection of early stage pancreatic cancer using 5-hydroxymethylcytosine signatures in circulating cell free DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/422675v1?rss=1</link>
<description><![CDATA[
Pancreatic cancers are typically diagnosed at late stage where disease prognosis is poor as exemplified by a 5-year survival rate of 8.2%. Earlier diagnosis would be beneficial by enabling surgical resection or earlier application of therapeutic regimens. We investigated the detection of pancreatic ductal adenocarcinoma (PDAC) in a non-invasive manner by interrogating changes in 5-hydroxymethylation cytosine status (5hmC) of circulating cell free DNA in the plasma of a PDAC cohort (n=51) in comparison with a non-cancer cohort (n=41). We found that 5hmC sites are enriched in a disease and stage specific manner in exons, 3UTRs and transcription termination sites. Our data show that 5hmC density is reduced in promoters and histone H3K4me3-associated sites with progressive disease suggesting increased transcriptional activity. 5hmC density is differentially represented in thousands of genes, and a stringently filtered set of the most significant genes points to biology related to pancreas (GATA4, GATA6, PROX1, ONECUT1) and/or cancer development (YAP1, TEAD1, PROX1, ONECUT1, ONECUT2, IGF1 and IGF2). Regularized regression models were built using 5hmC densities in statistically filtered genes or a comprehensive set of highly variable 5hmC counts in genes and performed with an AUC = 0.94-0.96 on training data. We were able to test the ability to classify PDAC and non-cancer samples with the Elastic net and Lasso models on two external pancreatic cancer 5hmC data sets and found validation performance to be AUC = 0.74-0.97. The findings suggest that 5hmC changes enable classification of PDAC patients with high fidelity and are worthy of further investigation on larger cohorts of patient samples.
]]></description>
<dc:creator>Collin, F.</dc:creator>
<dc:creator>Ning, Y.</dc:creator>
<dc:creator>Phillips, T.</dc:creator>
<dc:creator>McCarthy, E.</dc:creator>
<dc:creator>Scott, A.</dc:creator>
<dc:creator>Ellison, C.</dc:creator>
<dc:creator>Ku, C.-J.</dc:creator>
<dc:creator>Guler, G. D.</dc:creator>
<dc:creator>Chau, K.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Quake, S. R.</dc:creator>
<dc:creator>Levy, S.</dc:creator>
<dc:date>2018-09-20</dc:date>
<dc:identifier>doi:10.1101/422675</dc:identifier>
<dc:title><![CDATA[Detection of early stage pancreatic cancer using 5-hydroxymethylcytosine signatures in circulating cell free DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/424697v1?rss=1">
<title>
<![CDATA[
Cross-linking/Mass Spectrometry: A Community-Wide, Comparative Study Towards Establishing Best Practice Guidelines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/424697v1?rss=1</link>
<description><![CDATA[
The number of publications in the field of chemical cross-linking combined with mass spectrometry (XL-MS) to derive constraints for protein three-dimensional structure modeling and to probe protein-protein interactions has largely increased during the last years. As the technique is now becoming routine for in vitro and in vivo applications in proteomics and structural biology there is a pressing need to define protocols as well as data analysis and reporting formats that are generally accepted in the field and that have shown to lead to high-quality results. This first, community-based harmonization study on XL-MS is based on the results of 32 groups participating worldwide. The aim of this paper is to summarize the status quo of XL-MS and to compare and evaluate existing cross-linking strategies. From the results obtained, common protocols will be established. Our study serves as basis for establishing best practice guidelines in the field for conducting cross-linking experiments, performing data analysis, and reporting formats with the ultimate goal of assisting scientists to generate accurate and reproducible XL-MS results.
]]></description>
<dc:creator>Iacobucci, C.</dc:creator>
<dc:creator>Piotrowski, C.</dc:creator>
<dc:creator>Aebersold, R.</dc:creator>
<dc:creator>Amaral, B. C.</dc:creator>
<dc:creator>Andrews, P.</dc:creator>
<dc:creator>Borchers, C.</dc:creator>
<dc:creator>Brodie, N. I.</dc:creator>
<dc:creator>Bruce, J. E.</dc:creator>
<dc:creator>Chaignepain, S.</dc:creator>
<dc:creator>Chavez, J. D.</dc:creator>
<dc:creator>Claverol, S.</dc:creator>
<dc:creator>Cox, J.</dc:creator>
<dc:creator>Degliesposti, G.</dc:creator>
<dc:creator>Dong, M. Q.</dc:creator>
<dc:creator>Edinger, N.</dc:creator>
<dc:creator>Emanuelsson, C.</dc:creator>
<dc:creator>Gay, M.</dc:creator>
<dc:creator>Goetze, M.</dc:creator>
<dc:creator>Gozzo, F. C.</dc:creator>
<dc:creator>Gutierrez, C.</dc:creator>
<dc:creator>Haupt, C.</dc:creator>
<dc:creator>Heck, A. J. R.</dc:creator>
<dc:creator>Herzog, F.</dc:creator>
<dc:creator>Huang, L.</dc:creator>
<dc:creator>Hoopmann, M. R.</dc:creator>
<dc:creator>Kalisman, N.</dc:creator>
<dc:creator>Klykov, O.</dc:creator>
<dc:creator>Kukacka, Z.</dc:creator>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>MacCoss, M. J.</dc:creator>
<dc:creator>Mechtler, K.</dc:creator>
<dc:creator>Mesika, R.</dc:creator>
<dc:creator>Moritz, R. L.</dc:creator>
<dc:creator>Nagaraj, N.</dc:creator>
<dc:creator>Nesati, V.</dc:creator>
<dc:creator>Ninnis, R.</dc:creator>
<dc:creator>Novak, P.</dc:creator>
<dc:creator>O'Reilly, F. J.</dc:creator>
<dc:creator>Pelzing, M.</dc:creator>
<dc:creator>Petrotchenko, E.</dc:creator>
<dc:creator>Piersimoni, L.</dc:creator>
<dc:creator>Plasencia, M.</dc:creator>
<dc:creator>Pukala, T.</dc:creator>
<dc:creator>Rand,</dc:creator>
<dc:date>2018-09-26</dc:date>
<dc:identifier>doi:10.1101/424697</dc:identifier>
<dc:title><![CDATA[Cross-linking/Mass Spectrometry: A Community-Wide, Comparative Study Towards Establishing Best Practice Guidelines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/425231v1?rss=1">
<title>
<![CDATA[
phylogenize: a web tool to identify microbial genes underlying environment associations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/425231v1?rss=1</link>
<description><![CDATA[
SummaryPhylogenetic comparative methods are powerful but presently under-utilized ways to identify microbial genes underlying differences in community composition. These methods help to identify functionally important genes because they test for associations beyond those expected when related microbes occupy similar environments. We present phylogenize, a pipeline with web, QIIME2, and R interfaces that allows researchers to perform phylogenetic regression on 16S amplicon and shotgun sequencing data and to visualize results. phylogenize applies broadly to both host-associated and environmental microbiomes. Using Human Microbiome Project and Earth Microbiome Project data, we show that phylogenize draws similar conclusions from 16S versus shotgun sequencing and reveals both known and candidate pathways associated with host colonization.nnAvailabilityphylogenize is available at https://phylogenize.org and https://bitbucket.org/pbradz/phylogenize.nnContactkpollard@gladstone.ucsf.edu
]]></description>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2018-09-24</dc:date>
<dc:identifier>doi:10.1101/425231</dc:identifier>
<dc:title><![CDATA[phylogenize: a web tool to identify microbial genes underlying environment associations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/426338v1?rss=1">
<title>
<![CDATA[
FLASH: A next-generation CRISPR diagnostic for multiplexed detection of antimicrobial resistance sequences 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/426338v1?rss=1</link>
<description><![CDATA[
The growing prevalence of deadly microbes with resistance to previously life-saving drug therapies is a dire threat to human health. Detection of low abundance pathogen sequences remains a challenge for metagenomic Next Generation Sequencing (NGS). We introduce FLASH (Finding Low Abundance Sequences by Hybridization), a next-generation CRISPR/Cas9 diagnostic method that takes advantage of the efficiency, specificity and flexibility of Cas9 to enrich for a programmed set of sequences. FLASH-NGS achieves up to 5 orders of magnitude of enrichment and sub-attomolar gene detection with minimal background. We provide an open-source software tool (FLASHit) for guide RNA design. Here we applied it to detection of antimicrobial resistance genes in respiratory fluid and dried blood spots, but FLASH-NGS is applicable to all areas that rely on multiplex PCR.
]]></description>
<dc:creator>Quan, J.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>Kuchta, A.</dc:creator>
<dc:creator>Batson, J.</dc:creator>
<dc:creator>Teyssier, N.</dc:creator>
<dc:creator>Lyden, A.</dc:creator>
<dc:creator>Caldera, S.</dc:creator>
<dc:creator>McGeever, A.</dc:creator>
<dc:creator>Dimitrov, B.</dc:creator>
<dc:creator>King, R.</dc:creator>
<dc:creator>Wilheim, J.</dc:creator>
<dc:creator>Murphy, M.</dc:creator>
<dc:creator>Pesce Ares, L.</dc:creator>
<dc:creator>Travisano, K. A.</dc:creator>
<dc:creator>Sit, R.</dc:creator>
<dc:creator>Amato, R.</dc:creator>
<dc:creator>Mumbengegwi, D. R.</dc:creator>
<dc:creator>Smith, J. L.</dc:creator>
<dc:creator>Bennett, A.</dc:creator>
<dc:creator>Gosling, R.</dc:creator>
<dc:creator>Mourani, P. M.</dc:creator>
<dc:creator>Calfee, C. S.</dc:creator>
<dc:creator>Neff, N. F.</dc:creator>
<dc:creator>Chow, E. D.</dc:creator>
<dc:creator>Kim, P. S.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Crawford, E. D.</dc:creator>
<dc:date>2018-09-27</dc:date>
<dc:identifier>doi:10.1101/426338</dc:identifier>
<dc:title><![CDATA[FLASH: A next-generation CRISPR diagnostic for multiplexed detection of antimicrobial resistance sequences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/428250v1?rss=1">
<title>
<![CDATA[
The nuclear RNAi factor, NRDE2, prevents the accumulation of DNA damage during mitosis in stressful growth conditions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/428250v1?rss=1</link>
<description><![CDATA[
Organisms have evolved multiple mechanisms to prevent and repair DNA damage to protect the integrity of the genome, particularly under stressful conditions. Unrepaired DNA damage leads to genomic instability, aneuploidy, and an increased risk for cancer. Before the cell can divide, it must repair damaged DNA and it is thought that this process requires global silencing of most transcription. In C. elegans, NRDE-2, in complex with other nuclear factors and guided by small RNA, directs heterochromatin formation and transcriptional silencing of targeted genes. Additionally, when C. elegans are cultivated at high temperatures, NRDE-2 is required to maintain germ line immortality. However, the role of NRDE-2 in maintaining the physical integrity of the genome is not understood. We show here that loss of NRDE2 in either nematode or human cells induces the accumulation of DNA damage specifically under conditions of stress, such as cultivation at a high temperature in C. elegans or Aurora B Kinase oncogenic overexpression in the MCF10A epithelial breast cell line. In addition, we found that NRDE2 interacts with {beta}-actin in unstressed mammalian cells. This interaction is dramatically reduced upon DNA damage. The oligomerization state of nuclear actin alters its association with targets, which in turn, regulates their function. Monomeric nuclear actin binds to heterochromatin remodeling factors and transcriptional activators while filamentous actin has been implicated in DNA repair processes. Here we show that NRDE2 associates with actin only when DNA is intact and the bulk of nuclear actin is monomeric. Thus, NRDE2 may dissociate from actin when it becomes filamentous as a result of DNA damage. This implies that, NRDE2, in its role as a heterochromatin factor, binds to monomeric actin, protecting the genome from DNA damage in stressful conditions. In this way, heterochromatin factors may associate with the actin dependent DNA repair process to allow appropriate mitotic progression and maintain genomic integrity.
]]></description>
<dc:creator>Asuni, A.</dc:creator>
<dc:creator>Venkataramanan, S.</dc:creator>
<dc:creator>Suzuki, A.</dc:creator>
<dc:creator>Floor, S. N.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:creator>L'Etoile, N.</dc:creator>
<dc:date>2018-09-27</dc:date>
<dc:identifier>doi:10.1101/428250</dc:identifier>
<dc:title><![CDATA[The nuclear RNAi factor, NRDE2, prevents the accumulation of DNA damage during mitosis in stressful growth conditions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/432864v1?rss=1">
<title>
<![CDATA[
Light-based tuning of ligand half-life supports kinetic proofreading model of T cell activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/432864v1?rss=1</link>
<description><![CDATA[
T cells are thought to discriminate stimulatory versus non-stimulatory ligands by converting small changes in ligand binding half-life to large changes in cell activation. Such a kinetic proofreading model has been difficult to test directly, as existing methods of altering ligand binding half-life also change other potentially important biophysical parameters, most notably the stability of the receptor-ligand interaction under load. Here we develop an optogenetic approach to specifically tune the binding half-life of a light-responsive ligand to a chimeric antigen receptor without changing other binding parameters. By simultaneously manipulating binding half-life while controlling for receptor occupancy, we find that signaling is strongly gated by ligand binding half-life. Our results provide direct evidence of kinetic proofreading in ligand discrimination by T cells.nnOne Sentence SummaryDirect control of ligand binding half-life with light shows that lifetime, not occupancy, dominates T cell activation.
]]></description>
<dc:creator>Tischer, D.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:date>2018-10-02</dc:date>
<dc:identifier>doi:10.1101/432864</dc:identifier>
<dc:title><![CDATA[Light-based tuning of ligand half-life supports kinetic proofreading model of T cell activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/436907v1?rss=1">
<title>
<![CDATA[
Establishing a causal role for medial prefrontal cortex in reality monitoring 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/436907v1?rss=1</link>
<description><![CDATA[
Reality monitoring is defined as the ability to distinguish internally self-generated information from externally-derived information. Functional imaging studies have consistently found that the medial prefrontal cortex (mPFC) is a key brain region subserving reality monitoring. The aim of this study was to determine a causal role for mPFC in reality monitoring using navigated repetitive transcranial magnetic stimulation (nrTMS). In a subject-blinded sham-controlled crossover design, healthy individuals received either active or sham nrTMS targeting mPFC. Active modulation of mPFC using nrTMS at a frequency of 10 Hz, significantly improved identification of both self-generated and externally-derived information during reality monitoring, when compared to sham or baseline. Targeted excitatory modulation of mPFC also improved positive mood ratings, reduced negative mood ratings and increased overall alertness/arousal. These results establish optimal nrTMS dosing parameters that maximized tolerability/comfort and induced significant neuromodulatory effects in the mPFC target. Importantly, this is a proof-of-concept study that establishes the mPFC as a novel brain target that can be stimulated with nrTMS to causally impact both mood and higher-order reality monitoring.
]]></description>
<dc:creator>Subramaniam, K.</dc:creator>
<dc:creator>Kothare, H.</dc:creator>
<dc:creator>Hinkley, L.</dc:creator>
<dc:creator>Tarapore, P.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2018-10-07</dc:date>
<dc:identifier>doi:10.1101/436907</dc:identifier>
<dc:title><![CDATA[Establishing a causal role for medial prefrontal cortex in reality monitoring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/437244v1?rss=1">
<title>
<![CDATA[
Beta-band activity in medial prefrontal cortex predicts source memory encoding and retrieval accuracy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/437244v1?rss=1</link>
<description><![CDATA[
Reality monitoring is defined as the ability to distinguish internally self-generated information from externally-derived information. The medial prefrontal cortex (mPFC) is a key brain region subserving reality monitoring and has been shown to be activated specifically during the retrieval of self-generated information. However, it is unclear if mPFC is activated during the encoding of self-generated information into memory. If so, it is important to understand whether successful retrieval of self-generated information critically depends on enhanced neural activity within mPFC during initial encoding of this self-generated information.nnWe used magnetoencephalographic imaging (MEGI) to determine the timing and location of cortical activity during a reality-monitoring task involving self generated contextual source memory encoding and retrieval. We found both during encoding and retrieval of self-generated information, when compared to externally-derived information, mPFC showed significant task induced oscillatory power modulation in the beta-band. During initial encoding of self-generated information, greater mPFC beta-band power reductions occurred within a time window of -700ms to -500ms prior to vocalization, activity in mPFC that was not observed during encoding of externally-derived information. This mPFC activity during encoding of self-generated information predicted subsequent retrieval accuracy of self-generated information. Beta-band activity in mPFC was also observed during the initial retrieval of self-generated information within a time window of 300 to 500ms following stimulus onset and correlated with accurate retrieval performance of self-generated information. Together, these results further highlight the importance of mPFC in mediating the initial generation and awareness of participants internal thoughts.
]]></description>
<dc:creator>Subramaniam, K.</dc:creator>
<dc:creator>Hinkley, L.</dc:creator>
<dc:creator>Mizuiri, D.</dc:creator>
<dc:creator>Kothare, H.</dc:creator>
<dc:creator>Garrett, C.</dc:creator>
<dc:creator>Findlay, A.</dc:creator>
<dc:creator>Houde, J.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2018-10-07</dc:date>
<dc:identifier>doi:10.1101/437244</dc:identifier>
<dc:title><![CDATA[Beta-band activity in medial prefrontal cortex predicts source memory encoding and retrieval accuracy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/437368v1?rss=1">
<title>
<![CDATA[
Cell-type-specific resolution epigenetics without the need for cell sorting or single-cell biology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/437368v1?rss=1</link>
<description><![CDATA[
High costs and technical limitations of cell sorting and single-cell techniques currently restrict the collection of large-scale, cell-type-specific DNA methylation data. This, in turn, impedes our ability to tackle key biological questions that pertain to variation within a population, such as identification of disease-associated genes at a cell-type-specific resolution. Here, we show mathematically and empirically that cell-type-specific methylation levels of an individual can be learned from its tissue-level bulk data, conceptually emulating the case where the individual has been profiled with a single-cell resolution and then signals were aggregated in each cell population separately. Provided with this unprecedented way to perform powerful large-scale epigenetic studies with cell-type-specific resolution, we revisit previous studies with tissue-level bulk methylation and reveal novel associations with leukocyte composition in blood and with rheumatoid arthritis. For the latter, we further show consistency with validation data collected from sorted leukocyte sub-types. Corresponding software is available from: https://github.com/cozygene/TCA.
]]></description>
<dc:creator>Rahmani, E.</dc:creator>
<dc:creator>Schweiger, R.</dc:creator>
<dc:creator>Rhead, B.</dc:creator>
<dc:creator>Criswell, L. A.</dc:creator>
<dc:creator>Barcellos, L. F.</dc:creator>
<dc:creator>Eskin, E.</dc:creator>
<dc:creator>Rosset, S.</dc:creator>
<dc:creator>Sankararaman, S.</dc:creator>
<dc:creator>Halperin, E.</dc:creator>
<dc:date>2018-10-07</dc:date>
<dc:identifier>doi:10.1101/437368</dc:identifier>
<dc:title><![CDATA[Cell-type-specific resolution epigenetics without the need for cell sorting or single-cell biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/438705v1?rss=1">
<title>
<![CDATA[
Suppressor of Fused controls the proliferation of postnatal neural stem and precursor cells via a Gli3-dependent mechanism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/438705v1?rss=1</link>
<description><![CDATA[
The ventricular-subventricular zone (V-SVZ) of the forebrain is the source of neurogenic stem/precursor cells for adaptive and homeostatic needs throughout the life of most mammals. Here, we report that Suppressor of Fused (SUFU) plays a critical role in the establishment of the V-SVZ at early neonatal stages by controlling the proliferation of distinct subpopulations of stem/precursor cells. Conditional deletion of Sufu in radial glial progenitor cells (RGCs) at E13.5 resulted in a dramatic increase in the proliferation of Sox2+ Type B cells, as well as Gsx2+ ventral forebrain derived transit amplifying precursor cells (TACs). In contrast, we found a significant decrease in Tbr2+ dorsal forebrain derived TACs indicating that innate differences between dorsal and ventral forebrain derived Type B cells influence Sufu function. However, most precursors failed to survive and accumulated in the dorsal V-SVZ, demonstrating that precursors are unable to transition into functional differentiated progenies. These defects were accompanied by reduced Gli3 expression, yet despite reduced Gli3 levels, activation of Sonic hedgehog (Shh) signaling did not occur implying that the Sufu-Gli3 regulatory axis may influence other signaling pathways in the neonatal dorsal V-SVZ. These data suggest that Sufu plays a critical role in controlling Gli3 function in the establishment and survival of functional stem/precursor cells in the postnatal dorsal V-SVZ.nnSUMMARY STATEMENTConditional deletion of Sufu cause dramatic expansion of neural stem/precursor cells in the neonatal ventricular-subventricular (SVZ) zone. This defect occurs through a Gli3-dependent mechanism that results in the downregulation of Shh signaling.
]]></description>
<dc:creator>Gomez, H. G.</dc:creator>
<dc:creator>Castillo, J. G.</dc:creator>
<dc:creator>Noguchi, H.</dc:creator>
<dc:creator>Aguilar, D.</dc:creator>
<dc:creator>Pleasure, S. J.</dc:creator>
<dc:creator>Yabut, O. R.</dc:creator>
<dc:date>2018-10-09</dc:date>
<dc:identifier>doi:10.1101/438705</dc:identifier>
<dc:title><![CDATA[Suppressor of Fused controls the proliferation of postnatal neural stem and precursor cells via a Gli3-dependent mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/438911v1?rss=1">
<title>
<![CDATA[
Antiviral adaptive immunity and tolerance in the mosquito Aedes aegyti 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/438911v1?rss=1</link>
<description><![CDATA[
Mosquitoes spread pathogenic arboviruses while themselves tolerate infection. We here characterize an immunity pathway providing long-term antiviral protection and define how this pathway discriminates between self and non-self. Mosquitoes use viral RNAs to create viral derived cDNAs (vDNAs) central to the antiviral response. vDNA molecules are acquired through a process of reverse-transcription and recombination directed by endogenous retrotransposons. These vDNAs are thought to integrate in the host genome as endogenous viral elements (EVEs). Sequencing of pre-integrated vDNA revealed that the acquisition process exquisitely distinguishes viral from host RNA, providing one layer of self-nonself discrimination. Importantly, we show EVE-derived piRNAs have antiviral activity and are loaded onto Piwi4 to inhibit virus replication. In a second layer of self-non-self discrimination, Piwi4 preferentially loads EVE-derived piRNAs, discriminating against transposon-targeting piRNAs. Our findings define a fundamental virus-specific immunity pathway in mosquitoes that uses EVEs as a potent and specific antiviral transgenerational mechanism.
]]></description>
<dc:creator>Andino, R.</dc:creator>
<dc:creator>Tassetto, M.</dc:creator>
<dc:creator>Kunitomi, M.</dc:creator>
<dc:creator>Whitfield, Z. J.</dc:creator>
<dc:creator>Dolan, P. T.</dc:creator>
<dc:creator>Sanchez-Vargas, I.</dc:creator>
<dc:creator>Ribiero, I.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Olson, K. E.</dc:creator>
<dc:date>2018-10-09</dc:date>
<dc:identifier>doi:10.1101/438911</dc:identifier>
<dc:title><![CDATA[Antiviral adaptive immunity and tolerance in the mosquito Aedes aegyti]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/441295v1?rss=1">
<title>
<![CDATA[
Synthetic essentiality of metabolic regulator PDHK1 in PTEN-deficient cells and cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/441295v1?rss=1</link>
<description><![CDATA[
PTEN is a tumor suppressor that is often inactivated in cancer and possesses both lipid and protein phosphatase activities. We report the metabolic regulator PDHK1 (pyruvate dehydrogenase kinase1) is a synthetic-essential gene in PTEN-deficient cancer and normal cells. The predominant mechanism of PDHK1 regulation and dependency is the PTEN protein phosphatase dephosphorylates NF{kappa};B activating protein (NKAP) and limits NF{kappa}B activation to suppress expression of PDHK1, a NF{kappa}B target gene. Loss of the PTEN protein phosphatase upregulates PDHK1 to drive aerobic glycolysis and induce PDHK1 cellular dependence. PTEN-deficient human tumors harbor increased PDHK1, which is a biomarker of decreased patient survival, establishing clinical relevance. This study uncovers a PTEN-regulated signaling pathway and reveals PDHK1 as a potential target in PTEN-deficient cancers.nnSIGNIFICANCEThe tumor suppressor PTEN is widely inactivated in cancers and tumor syndromes. PTEN antagonizes PI3K/AKT signaling via its lipid phosphatase activity. The modest success of PI3K/AKT inhibition in PTEN-deficient cancer patients provides rationale for identifying other vulnerabilities in PTEN-deficient cancers to improve clinical outcomes. We show that PTEN-deficient cells are uniquely sensitive to PDHK1 inhibition. PTEN and PDHK1 co-suppression reduced colony formation and induced cell death in vitro and tumor regression in vivo. PDHK1 levels were high in PTEN-deficient patient tumors and associated with inferior patient survival, establishing clinical relevance. Our study identifies a PTEN-regulated signaling pathway linking the PTEN protein phosphatase to the metabolic regulator PDHK1 and provides a mechanistic basis for PDHK1 targeting in PTEN-deficient cancers.
]]></description>
<dc:creator>Chatterjee, N.</dc:creator>
<dc:creator>Pazarentzos, E.</dc:creator>
<dc:creator>Hrustanovic, G.</dc:creator>
<dc:creator>Lin, L.</dc:creator>
<dc:creator>Verschueren, E.</dc:creator>
<dc:creator>Johnson, J. R.</dc:creator>
<dc:creator>Hofree, M.</dc:creator>
<dc:creator>Yan, J. J.</dc:creator>
<dc:creator>Olivas, V.</dc:creator>
<dc:creator>Newton, B. W.</dc:creator>
<dc:creator>Dollen, J. V.</dc:creator>
<dc:creator>Earnshaw, C. H.</dc:creator>
<dc:creator>Flanagan, J.</dc:creator>
<dc:creator>Chan, E.</dc:creator>
<dc:creator>Asthana, S.</dc:creator>
<dc:creator>Ideker, T.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Mayekar, M. K.</dc:creator>
<dc:creator>Suzuki, J.</dc:creator>
<dc:creator>Barad, B.</dc:creator>
<dc:creator>Kirichok, Y.</dc:creator>
<dc:creator>Fraser, J.</dc:creator>
<dc:creator>Weiss, W. A.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Tulpule, A.</dc:creator>
<dc:creator>Sabnis, A. J.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:date>2018-10-11</dc:date>
<dc:identifier>doi:10.1101/441295</dc:identifier>
<dc:title><![CDATA[Synthetic essentiality of metabolic regulator PDHK1 in PTEN-deficient cells and cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/441352v1?rss=1">
<title>
<![CDATA[
Cyclin B2 is required for progression through meiosis in mouse oocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/441352v1?rss=1</link>
<description><![CDATA[
Cyclins associate with CDK1 to generate the M-phase-promoting factor (MPF) essential for progression through mitosis and meiosis. Previous studies concluded that CCNB2 is dispensable for cell cycle progression. Given our findings of high translation rates of CcnB2 mRNA in prophase-arrested oocytes, we have reevaluated its role during meiosis. CcnB2-/- oocytes undergo delayed germinal vesicle breakdown followed by a defective M-phase due to reduced pre-MPF activity. This disrupted maturation is associated with compromised CcnB1 and Mos mRNA translation and delayed spindle assembly. Given these defects, a significant population of oocytes fail to complete meiosis I because SAC remains activated and APC function is inhibited. In vivo, CCNB2 depletion leads to decreased oocyte developmental competence, compromised fecundity, and premature ovarian failure. These findings demonstrate that CCNB2 is required to assemble sufficient pre-MPF for timely meiosis reentry and progression. Although endogenous cyclins cannot compensate, overexpression of CCNB1 rescues the meiotic phenotypes, demonstrating similar molecular properties but divergent modes of regulation of these cyclins.
]]></description>
<dc:creator>Daldello, E. M.</dc:creator>
<dc:creator>Luong, X. G.</dc:creator>
<dc:creator>Yang, C.-R.</dc:creator>
<dc:creator>Huhn, J.</dc:creator>
<dc:creator>Conti, M.</dc:creator>
<dc:date>2018-10-12</dc:date>
<dc:identifier>doi:10.1101/441352</dc:identifier>
<dc:title><![CDATA[Cyclin B2 is required for progression through meiosis in mouse oocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/445833v1?rss=1">
<title>
<![CDATA[
The glucocorticoid receptor ligand-binding domain confers drug-gated protein regulation in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/445833v1?rss=1</link>
<description><![CDATA[
Controlling protein activity and localization is a key tool in modern biology. Mammalian steroid receptor ligand-binding domains (LBDs) fusions have been used in a range of organisms and cell-types to inactivate proteins of interest until the cognate steroid ligand is applied. Here, we demonstrate that the glucocorticoid receptor LBD confers ligand-gated control of a heterologous gene expression system (Q system) and the DAF-16 transcription factor in C. elegans. These experiments demonstrate provide a powerful tool for temporal control of protein activity, and will bolster existing tools used to modulate gene expression and protein activity in this animal.
]]></description>
<dc:creator>Monsalve, G. C.</dc:creator>
<dc:creator>Yamamoto, K. R.</dc:creator>
<dc:creator>Ward, J. D.</dc:creator>
<dc:date>2018-10-17</dc:date>
<dc:identifier>doi:10.1101/445833</dc:identifier>
<dc:title><![CDATA[The glucocorticoid receptor ligand-binding domain confers drug-gated protein regulation in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/446492v1?rss=1">
<title>
<![CDATA[
Reverse GWAS: Using Genetics to Identify and Model Phenotypic Subtypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/446492v1?rss=1</link>
<description><![CDATA[
Recent and classical work has revealed biologically and medically significant subtypes in complex diseases and traits. However, relevant subtypes are often unknown, unmeasured, or actively debated, making automatic statistical approaches to subtype definition particularly valuable. We propose reverse GWAS (RGWAS) to identify and validate subtypes using genetics and multiple traits: while GWAS seeks the genetic basis of a given trait, RGWAS seeks to define trait subtypes with distinct genetic bases. Unlike existing approaches relying on off-the-shelf clustering methods, RGWAS uses a bespoke decomposition, MFMR, to model covariates, binary traits, and population structure. We use extensive simulations to show these features can be crucial for power and calibration. We validate RGWAS in practice by recovering known stress subtypes in major depressive disorder. We then show the utility of RGWAS by identifying three novel subtypes of metabolic traits. We biologically validate these metabolic subtypes with SNP-level tests and a novel polygenic test: the former recover known metabolic GxE SNPs; the latter suggests genetic heterogeneity may explain substantial missing heritability. Crucially, statins, which are widely prescribed and theorized to increase diabetes risk, have opposing effects on blood glucose across metabolic subtypes, suggesting potential have potential translational value.nnAuthor summaryComplex diseases depend on interactions between many known and unknown genetic and environmental factors. However, most studies aggregate these strata and test for associations on average across samples, though biological factors and medical interventions can have dramatically different effects on different people. Further, more-sophisticated models are often infeasible because relevant sources of heterogeneity are not generally known a priori. We introduce Reverse GWAS to simultaneously split samples into homogeneoues subtypes and to learn differences in genetic or treatment effects between subtypes. Unlike existing approaches to computational subtype identification using high-dimensional trait data, RGWAS accounts for covariates, binary disease traits and, especially, population structure; these features are each invaluable in extensive simulations. We validate RGWAS by recovering known genetic subtypes of major depression. We demonstrate RGWAS is practically useful in a metabolic study, finding three novel subtypes with both SNP- and polygenic-level heterogeneity. Importantly, RGWAS can uncover differential treatment response: for example, we show that statin, a common drug and potential type 2 diabetes risk factor, may have opposing subtype-specific effects on blood glucose.
]]></description>
<dc:creator>Dahl, A.</dc:creator>
<dc:creator>Cai, N.</dc:creator>
<dc:creator>Ko, A.</dc:creator>
<dc:creator>Laakso, M.</dc:creator>
<dc:creator>Pajukanta, P.</dc:creator>
<dc:creator>Flint, J.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:date>2018-10-17</dc:date>
<dc:identifier>doi:10.1101/446492</dc:identifier>
<dc:title><![CDATA[Reverse GWAS: Using Genetics to Identify and Model Phenotypic Subtypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/451393v1?rss=1">
<title>
<![CDATA[
LC3 and STRAP regulate actin filament assembly by JMY during autophagosome formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/451393v1?rss=1</link>
<description><![CDATA[
During autophagy actin filament networks move and remodel cellular membranes to form autophagosomes that enclose and metabolize cytoplasmic contents. Two actin regulators, WHAMM and JMY, participate in autophagosome formation, but the signals linking autophagy to actin assembly are poorly understood. We show that, in non-starved cells, cytoplasmic JMY co-localizes with STRAP, a regulator of JMYs nuclear functions, on non-motile vesicles with no associated actin networks. Upon starvation, JMY shifts to motile, LC3-containing membranes that move on actin comet tails. LC3 enhances JMYs de novo actin nucleation activity via a cryptic actin-binding sequence near JMYs N-terminus, and STRAP inhibits JMYs ability to nucleate actin and activate the Arp2/3 complex. Cytoplasmic STRAP negatively regulates autophagy. Finally, we use purified proteins to reconstitute LC3- and JMY-dependent actin network formation on membranes, and inhibition of network formation by STRAP. We conclude that LC3 and STRAP regulate JMYs actin assembly activities in trans during autophagy.nneTOC BlurbThe actin regulator JMY creates filament networks that move membranes during autophagy. We find that, in unstarved cells, JMY is inhibited by interaction with the STRAP protein, but upon starvation JMY is recruited away from STRAP and activated by LC3.
]]></description>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Mullins, R. D.</dc:creator>
<dc:date>2018-10-24</dc:date>
<dc:identifier>doi:10.1101/451393</dc:identifier>
<dc:title><![CDATA[LC3 and STRAP regulate actin filament assembly by JMY during autophagosome formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/452334v1?rss=1">
<title>
<![CDATA[
Insertion sequences drive the emergence of a highly adapted human pathogen 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/452334v1?rss=1</link>
<description><![CDATA[
Taxonomic outliers of Pseudomonas aeruginosa of environmental origin have recently emerged as infectious for humans. Here we present the first genome-wide analysis of an isolate that caused fatal hemorrhagic pneumonia. We demonstrate that, in two sequential clones, CLJ1 and CLJ3, recovered from a patient with chronic pulmonary disease, insertion of a mobile genetic element into the P. aeruginosa chromosome affected major virulence-associated phenotypes and led to increased resistance to antibiotics used to treat the patient. Comparative proteome and transcriptome analyses revealed that this insertion sequence, ISL3, disrupted genes encoding flagellar components, type IV pili, O-specific antigens, translesion polymerase and enzymes producing hydrogen cyanide. CLJ3 possessed seven fold more IS insertions than CLJ1, some modifying its susceptibility to antibiotics by disrupting the genes for the outer-membrane porin OprD and the regulator of {beta}-lactamase expression AmpD. In the Galleria mellonella larvae model, the two strains displayed different levels of virulence, with CLJ1 being highly pathogenic. This work reveals ISs as major players in enhancing the pathogenic potential of a P. aeruginosa taxonomic outlier by modulating both, the virulence and the resistance to antimicrobials, and explains the ability of this bacterium to adapt from the environment to a human host.
]]></description>
<dc:creator>Sentausa, E.</dc:creator>
<dc:creator>Basso, P.</dc:creator>
<dc:creator>Berry, A.</dc:creator>
<dc:creator>Adrait, A.</dc:creator>
<dc:creator>Bellement, G.</dc:creator>
<dc:creator>Coute, Y.</dc:creator>
<dc:creator>Lory, S.</dc:creator>
<dc:creator>Elsen, S.</dc:creator>
<dc:creator>Attree, I.</dc:creator>
<dc:date>2018-10-25</dc:date>
<dc:identifier>doi:10.1101/452334</dc:identifier>
<dc:title><![CDATA[Insertion sequences drive the emergence of a highly adapted human pathogen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/452417v1?rss=1">
<title>
<![CDATA[
Building Key Populations HIV Treatment Cascades in Data-Scarce Environments: Towards a participatory stakeholder methodology for cascades construction, adoption, and utilization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/452417v1?rss=1</link>
<description><![CDATA[
IntroductionRecent HIV key populations (KP) surveillance studies in South Africa, including female sex workers (FSW) and men who have sex with men (MSM), demonstrate the disproportionate burden of HIV they bear compared to the general population. The national response for KP has lagged due to relatively scarce KP data focused narrowly on urban areas. We adopted a participatory data triangulation approach with stakeholders to overcome the challenges of KP program planning in KP data-scarce districts. Here we describe our methodology for achieving consensus on population size estimates (PSE) and treatment cascade indicator estimates derived from FSW and MSM surveillance data and applied across the country.nnMethodsThe South African National AIDS Council (SANAC) convened the group; technical advisors from the University of California San Francisco (UCSF) facilitated; and stakeholders from government, non-government, academic, and KP-led advocacy organizations involved in program implementation and research provided input through three in-person meetings covering four phases of work: surveillance data analysis; cascades data extraction; presentation for feedback; and data extrapolation.nnResultsTechnical advisors presented eight cascades (three FSW, five MSM) to stakeholders, recommending data-informed extrapolation factors for each population. Stakeholders adopted recommendations by consensus with few adjustments. FSW cascades displayed high awareness of HIV status and steep breakpoints towards ART uptake; MSM cascades displayed less HIV status awareness, but relatively good ART uptake, with metropolitan areas displaying better uptake than rural districts.nnConclusionThe participatory process enabled KP stakeholders to vet disparate data sources against programmatic experience and recommend consistency in cascades data; participatory triangulation of additional surveillance and program data will follow. The considerable time and resource investments in this process had downstream benefits, including consistency in sub-national HIV implementation plans. We recommend this consensus-based approach as a transparent, consistent, and sound methodology for cascades construction in KP data-scarce environments.
]]></description>
<dc:creator>Lane, T.</dc:creator>
<dc:creator>Grasso, M.</dc:creator>
<dc:creator>Scheibe, A.</dc:creator>
<dc:creator>Liu, G.</dc:creator>
<dc:creator>Marr, A.</dc:creator>
<dc:creator>Murangandi, P.</dc:creator>
<dc:creator>Aynalem, G.</dc:creator>
<dc:creator>Slabbert, M.</dc:creator>
<dc:creator>Malaka, L.</dc:creator>
<dc:creator>Isdahl, Z.</dc:creator>
<dc:creator>Osmand, T.</dc:creator>
<dc:creator>Nadol, P.</dc:creator>
<dc:date>2018-10-24</dc:date>
<dc:identifier>doi:10.1101/452417</dc:identifier>
<dc:title><![CDATA[Building Key Populations HIV Treatment Cascades in Data-Scarce Environments: Towards a participatory stakeholder methodology for cascades construction, adoption, and utilization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/457119v1?rss=1">
<title>
<![CDATA[
Cell-extrinsic mechanical forces restore neutrophil polarization in the absence of branched actin assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/457119v1?rss=1</link>
<description><![CDATA[
Migratory cells use distinct motility modes to navigate different microenvironments, but it is unclear whether these modes rely on the same core set of polarity components. To investigate this, we disrupted Arp2/3 and WAVE complex, which assemble branched actin networks that are essential for neutrophil polarity and motility in standard adherent conditions. Surprisingly, confinement rescues polarity and movement of neutrophils lacking these components, revealing a processive bleb-based protrusion program that is mechanistically distinct from the branched actin-based protrusion program but shares some of the same core components and underlying molecular logic. We further find that the restriction of protrusion growth to one site does not always respond to membrane tension directly, as previously thought, but may rely on closely linked properties such as local membrane curvature. Our work reveals a hidden circuit for neutrophil polarity and indicates that cells have distinct molecular mechanisms for polarization that dominate in different microenvironments.
]]></description>
<dc:creator>Graziano, B. R.</dc:creator>
<dc:creator>Town, J. P.</dc:creator>
<dc:creator>Nagy, T. L.</dc:creator>
<dc:creator>Fosnaric, M.</dc:creator>
<dc:creator>Penic, S.</dc:creator>
<dc:creator>Iglic, A.</dc:creator>
<dc:creator>Kralj-Iglic, V.</dc:creator>
<dc:creator>Gov, N.</dc:creator>
<dc:creator>Diz-Munoz, A.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:date>2018-10-30</dc:date>
<dc:identifier>doi:10.1101/457119</dc:identifier>
<dc:title><![CDATA[Cell-extrinsic mechanical forces restore neutrophil polarization in the absence of branched actin assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/461285v1?rss=1">
<title>
<![CDATA[
Real-time projections of epidemic transmission and estimation of vaccination impact during an Ebola virus disease outbreak in the Eastern region of the Democratic Republic of Congo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/461285v1?rss=1</link>
<description><![CDATA[
BackgroundAs of October 12, 2018, 211 cases of Ebola virus disease (EVD) were reported in North Kivu Province, Democratic Republic of Congo. Since the beginning of October the outbreak has largely shifted into regions in which active armed conflict is occurring, and in which EVD cases and their contacts are difficult for health workers to reach. We used available data on the current outbreak with case-count time series from prior outbreaks to project the short-term and long-term course of the outbreak.nnMethodsFor short and long term projections we modeled Ebola virus transmission using a stochastic branching process that assumes gradually quenching transmission estimated from past EVD outbreaks, with outbreak trajectories conditioned on agreement with the course of the current outbreak, and with multiple levels of vaccination coverage. We used a negative binomial autoregression for short-term projections, a Theil-Sen regression model for final sizes, and a baseline minimum-information projection using Gotts law to construct an ensemble of forecasts to be compared and recorded for future evaluation against final outcomes. From August 20 to October 13, short-term model projections were validated against actual case counts.nnResultsDuring validation of short-term projections, from one week to four weeks, we found models consistently scored higher on shorter-term forecasts. Based on case counts as of October 13, the stochastic model projected a median case count of 226 cases by October 27 (95% prediction interval: 205-268) and 245 cases by November 10 (95% prediction interval: 208-315), while the auto-regression model projects median case counts of 240 (95% prediction interval: 215-307) and 259 (95% prediction interval: 216-395) cases for those dates, respectively. Projected median final counts range from 274 to 421. Except for Gotts law, the projected probability of an outbreak comparable to 2013-2016 is exceedingly small. The stochastic model estimates that vaccine coverage in this outbreak is lower than reported in its trial setting in Sierra Leone.nnConclusionsBased on our projections we believe that the epidemic had not yet peaked at the time of these estimates, though a trajectory on the scale of the West African outbreak is exceedingly improbable. Validating our models in real time allowed us to generate more accurate short-term forecasts, and this process may provide a useful roadmap for real-time short-term forecasting. We estimate that transmission rates are higher than would be seen under target levels of 62% coverage due to contact tracing and vaccination, and this model estimate may offer a surrogate indicator for the outbreak response challenges.
]]></description>
<dc:creator>Worden, L.</dc:creator>
<dc:creator>Wannier, R.</dc:creator>
<dc:creator>Hoff, N. A.</dc:creator>
<dc:creator>Musene, K.</dc:creator>
<dc:creator>Selo, B.</dc:creator>
<dc:creator>Mossoko, M.</dc:creator>
<dc:creator>Okitolonda-Wemakoy, E.</dc:creator>
<dc:creator>Muyembe-Tamfum, J. J.</dc:creator>
<dc:creator>Rutherford, G.</dc:creator>
<dc:creator>Lietman, T. M.</dc:creator>
<dc:creator>Rimoin, A. W.</dc:creator>
<dc:creator>Porco, T. C.</dc:creator>
<dc:creator>Kelly, J. D.</dc:creator>
<dc:date>2018-11-05</dc:date>
<dc:identifier>doi:10.1101/461285</dc:identifier>
<dc:title><![CDATA[Real-time projections of epidemic transmission and estimation of vaccination impact during an Ebola virus disease outbreak in the Eastern region of the Democratic Republic of Congo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/468140v1?rss=1">
<title>
<![CDATA[
Matrix linear models for high-throughput chemical genetic screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/468140v1?rss=1</link>
<description><![CDATA[
We develop a flexible and computationally efficient approach for analysing high throughput chemical genetic screens. In such screens, a library of genetic mutants is phenotyped in a large number of stresses. The goal is to detect interactions between genes and stresses. Typically, this is achieved by grouping the mutants and stresses into categories, and performing modified t-tests for each combination. This approach does not have a natural extension if mutants or stresses have quantitative or non-overlapping annotations (eg. if conditions have doses, or a mutant falls into more than one category simultaneously). We develop a matrix linear model framework that allows us to model relationships between mutants and conditions in a simple, yet flexible multivariate framework. It encodes both categorical and continuous relationships to enhance detection of associations. To handle large datasets, we develop a fast estimation approach that takes advantage of the structure of matrix linear models. We evaluate our methods performance in simulations and in an E. coli chemical genetic screen, comparing it with an existing univariate approach based on modified t-tests. We show that matrix linear models perform slightly better than the univariate approach when mutants and conditions are classified in non-overlapping categories, and substantially better when conditions can be ordered in dosage categories. Our approach is much faster computationally and is scalable to larger datasets. It is an attractive alternative to current methods, and provides a natural framework extensible to larger, and more complex chemical genetic screens. A Julia implementation of matrix linear models and the code used for the analysis in this paper can be found at https://bitbucket.org/jwliang/mlm_packages and https://bitbucket.org/jwliang/mlm_gs_supplement, respectively.
]]></description>
<dc:creator>Liang, J. W.</dc:creator>
<dc:creator>Nichols, R. J.</dc:creator>
<dc:creator>Sen, S.</dc:creator>
<dc:date>2018-11-11</dc:date>
<dc:identifier>doi:10.1101/468140</dc:identifier>
<dc:title><![CDATA[Matrix linear models for high-throughput chemical genetic screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/469635v1?rss=1">
<title>
<![CDATA[
Unbiased detection of CRISPR off-targets in vivo using DISCOVER-Seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/469635v1?rss=1</link>
<description><![CDATA[
Genome editing using nucleases such as CRISPR-Cas induces programmable DNA damage at a target genomic site but can also affect off-target sites. Here, we develop a powerful, sensitive assay for the unbiased identification of off-target sites that we term DISCOVER-Seq. This approach takes advantage of the recruitment of endogenous DNA repair factors for genome-wide identification of Cas-induced double-strand breaks. One such factor, MRE11, is recruited precisely to double-strand breaks, enabling molecular characterization of nuclease cut sites with single-base resolution. DISCOVER-Seq detects off-targets in cellular models and in vivo upon adenoviral gene editing of mouse livers, paving the way for real-time off-target discovery during therapeutic gene editing. DISCOVER-Seq is furthermore applicable to multiple types of Cas nucleases and provides an unprecedented view of events that precede repair of the affected sites.
]]></description>
<dc:creator>Wienert, B.</dc:creator>
<dc:creator>Wyman, S. K.</dc:creator>
<dc:creator>Richardson, C. D.</dc:creator>
<dc:creator>Yeh, C. D.</dc:creator>
<dc:creator>Akcakaya, P.</dc:creator>
<dc:creator>Porritt, M. J.</dc:creator>
<dc:creator>Morlock, M.</dc:creator>
<dc:creator>Vu, J. T.</dc:creator>
<dc:creator>Kazane, K. R.</dc:creator>
<dc:creator>Watry, H. L.</dc:creator>
<dc:creator>Judge, L. M.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:creator>Maresca, M.</dc:creator>
<dc:creator>Corn, J. E.</dc:creator>
<dc:date>2018-11-14</dc:date>
<dc:identifier>doi:10.1101/469635</dc:identifier>
<dc:title><![CDATA[Unbiased detection of CRISPR off-targets in vivo using DISCOVER-Seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/469700v1?rss=1">
<title>
<![CDATA[
TFF3 is a ligand for LINGO2 that de-represses EGFR to control disease outcome during colitis and gastrointestinal nematode infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/469700v1?rss=1</link>
<description><![CDATA[
Intestinal epithelial cells (IEC) comprise diverse lineages that serve distinct roles necessary for regulation of nutrient absorption, regeneration, immunity, and homeostasis1,2. Goblet cells secrete Trefoil factor 3 (TFF3) to maintain mucus viscosity and drive mucosal healing by inhibiting cell death and influencing tight junction protein expression3. However, whether TFF3 signaling relies upon conventional ligand-receptor interactions has been unclear for decades. This study demonstrates that the orphan transmembrane protein leucine rich repeat receptor and nogo-interacting protein 2 (LINGO2) immunoprecipitates with TFF3, that LINGO2 and TFF3 co-localize at the IEC cell surface, and that TFF3/LINGO2 interactions block IEC apoptosis. Loss of function studies show that TFF3-driven STAT3 and EGFR activation are both LINGO2 dependent. Importantly, we demonstrate that TFF3 disrupts LINGO2/EGFR interactions that normally restrict EGFR activity, resulting in enhanced EGFR signaling. Excessive EGFR activation in Lingo2 gene deficient mice exacerbates colitic disease and accelerates host resistance to parasitic nematodes, whereas TFF3 deficiency results in host susceptibility. Thus, our data demonstrating that TFF3 functions through a previously unrecognized ligand-receptor interaction with LINGO2 to de-repress LINGO2-dependent inhibition of EGFR activation provides a novel conceptual framework explaining how TFF3-mediates mucosal wound healing through enhanced activation of the EGFR pathway.
]]></description>
<dc:creator>Ji, Y.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Hung, L. Y.</dc:creator>
<dc:creator>Young, T.</dc:creator>
<dc:creator>Herbine, K.</dc:creator>
<dc:creator>Oniskey, T.</dc:creator>
<dc:creator>Pastore, C.</dc:creator>
<dc:creator>Nieves, W.</dc:creator>
<dc:creator>Somsouk, M.</dc:creator>
<dc:creator>Herbert, D.</dc:creator>
<dc:date>2018-11-13</dc:date>
<dc:identifier>doi:10.1101/469700</dc:identifier>
<dc:title><![CDATA[TFF3 is a ligand for LINGO2 that de-represses EGFR to control disease outcome during colitis and gastrointestinal nematode infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/472787v1?rss=1">
<title>
<![CDATA[
Transient activation of the UPRER is an essential step in the acquisition of pluripotency during reprogramming 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/472787v1?rss=1</link>
<description><![CDATA[
Somatic cells can be reprogrammed into pluripotent stem cells by the forced expression of the OCT4, SOX2, KLF4 and c-MYC transcription factors. This process requires the reshaping of not only epigenetic landscapes, but the global remodeling of cell identity, structure, and function including such basic processes of metabolism and organelle form and function. Cellular reprogramming is a stochastic process with only a marginally measureable fraction of cells successfully crossing these, and many other, cellular epitomes to acquire the fully pluripotent state. We hypothesize that this variation is due, in part, by variable regulation of the proteostasis network and its influence upon the protein folding environment within cells and their organelles upon the remodeling process. We find that the endoplasmic reticulum unfolded protein response (UPRER), the heat-shock response (HSR) and the mitochondrial unfolded protein response (UPRmt), which monitor and ensure the quality of the proteome of, respectively, the ER, the cytosol and the mitochondria during stress, are activated during cellular reprogramming. Particularly, we find that the UPRER is essential for reprograming, and ectopic, transient activation of the UPRER, either genetically or pharmacologically, enhances the success of cells to reach a pluripotent state. Finally, and most revealing, we find that stochastic activation of the UPRER can predict the reprogramming efficiency of naive cells. The results of these experiments indicate that the low efficiency and stochasticity of cellular reprogramming is partly the result of the inability to initiate a proper ER stress response for remodeling of the ER and its proteome during the reprogramming process. The results reported here display only one aspect of the proteostasis network and suggest that proper regulation of many more components of this network might be essential to acquire the pluripotent state.
]]></description>
<dc:creator>Simic, M.</dc:creator>
<dc:creator>Moehle, E.</dc:creator>
<dc:creator>Schinzel, R.</dc:creator>
<dc:creator>Lorbeer, F.</dc:creator>
<dc:creator>Jullie, D.</dc:creator>
<dc:creator>Halloran, J.</dc:creator>
<dc:creator>Heydari, K.</dc:creator>
<dc:creator>Sanchez, M.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:creator>Dillin, A.</dc:creator>
<dc:date>2018-11-17</dc:date>
<dc:identifier>doi:10.1101/472787</dc:identifier>
<dc:title><![CDATA[Transient activation of the UPRER is an essential step in the acquisition of pluripotency during reprogramming]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/473082v1?rss=1">
<title>
<![CDATA[
Tracing diagnosis trajectories over millions of inpatients reveal an unexpected association between schizophrenia and rhabdomyolysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/473082v1?rss=1</link>
<description><![CDATA[
While it has been technically feasible to create longitudinal representations of individual health at a nationwide scale, the use of these techniques to identify novel disease associations for the risk stratification of patients has had limited success. Here, we created a large-scale US longitudinal disease network of traced readmission patterns (i.e., disease trajectories), merging data from over 10.4 million inpatients from 350 California hospitals through the Healthcare Cost and Utilization Project between 1980 and 2010. We were able to create longitudinal representations of disease progression mapping over 300 common diseases, including the well-known complication of heart failure after acute myocardial infarction. Surprisingly, out of these generated disease trajectories, we discovered an unknown association between schizophrenia, a chronic mental disorder, and rhabdomyolysis, a rare disease of muscle breakdown. It was found that 92 of 3674 patients (2.5%) with schizophrenia were readmitted for rhabdomyolysis (relative risk, 2.21 [1.80-2.71, confidence interval = 0.95] P-value 9.54E-15), which has a general population incidence of 1 in 10,000. We validated this association using independent electronic health records from over 830,000 patients treated over seven years at the University of California, San Francisco (UCSF) medical center. A case review of 29 patients at UCSF who were treated for schizophrenia and who went on to develop rhabdomyolysis demonstrated that the majority of cases (62%) are idiopathic, which suggests a biological connection between these two diseases. Together, these findings demonstrate the power of using public disease registries in combination with electronic medical records to discover novel disease associations.nnOne Sentence SummaryBased on the longitudinal health records from millions of California inpatient discharges, we created a temporal network that enabled us to understand statewide patterns of hospital readmissions, which led to the novel finding that hospitalization for schizophrenia is significantly associated with rehospitalization for rhabdomyolysis.
]]></description>
<dc:creator>Paik, H.</dc:creator>
<dc:creator>Kan, M. J.</dc:creator>
<dc:creator>Rappoport, N.</dc:creator>
<dc:creator>Hadley, D.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Chen, B.</dc:creator>
<dc:creator>Manber, U.</dc:creator>
<dc:creator>Cho, S. B.</dc:creator>
<dc:creator>Butte, A. J.</dc:creator>
<dc:date>2018-11-19</dc:date>
<dc:identifier>doi:10.1101/473082</dc:identifier>
<dc:title><![CDATA[Tracing diagnosis trajectories over millions of inpatients reveal an unexpected association between schizophrenia and rhabdomyolysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/473363v1?rss=1">
<title>
<![CDATA[
Age but not disease progression defines CD4+ and CD8+ T stem cell memory levels in human retroviral infections: contrasting effects of HTLV-1 and HIV-1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/473363v1?rss=1</link>
<description><![CDATA[
BackgroundHuman CD4+ and CD8+ stem cell memory T cells (TSCM) represent a minor fraction of circulating lymphocytes characterized by stemness and long-term in vivo persistence. CD4+ TSCM are preferentially infected and constitute a reservoir for HIV-1, whereas CD8+ TSCM appear to play a protective role. However, little is known about CD4+ and CD8+ TSCM in the only other human pathogenic retroviral infection, human T-cell leukemia virus type 1 (HTLV-1). HTLV-1 is the etiological agent of both Adult T-cell Leukemia (ATL) and HTLV-1 associated myelopathy/tropical spastic paraperesis (HAM/TSP), a neuroinflammatory disorder. In ATL, CD4+ TSCM cells were identified as the hierarchical leukemic stem cell, but data in HAM/TSP are lacking. Age is a major risk factor for both ATL and HAM/TSP, as both diseases generally manifest several decades after infection. Therefore, we explored a possible link between TSCM, age and disease status in human retroviral infections in a cross-sectional study, using multiparametric flow cytometry.nnResultsWe found that CD4+ or CD8+ TSCM levels (quantified as CD3+CD45RA+CD45RO- CD27+CCR7+Fashi) do not differ between healthy controls and untreated HTLV-1 infected individuals with and without neuroinflammatory disorder. However, we found both TSCM as well as CD8+ TSCM significantly accumulated with age, resulting in a >400% increase in elderly HTLV-1 infected individuals (>60 years). A significant correlation between age and TSCM signature genes was validated at the transcriptome level in an independent cohort. CD8+ but not CD4+ TSCM were significantly decreased in untreated HIV-1 infection. Unexpectedly, CD8+ TSCM recovery upon successful antiretroviral treatment was essentially complete (92.2{+/-}11.0%) in younger (<45 years) individuals, but significantly lower (37.3{+/-}6.1%) in older (>45 years) individuals (p=0.0003).nnConclusionIn HTLV-1 infection, an age-dependent accumulation of CD4+ and CD8+ TSCM points towards a possible protective role of CD8 TSCM in the elderly against leukemic but not neuroinflammatory disease. HIV-1-infected individuals lose their ability to restore CD8+ TSCM levels upon successful antiretroviral therapy at later age (>45 years), which might eventually lead to immunological failure and decreased vaccine efficacy.
]]></description>
<dc:creator>Menezes, S. M.</dc:creator>
<dc:creator>Leal, F. E.</dc:creator>
<dc:creator>Pereira Ribeiro, S.</dc:creator>
<dc:creator>Dierckx, T.</dc:creator>
<dc:creator>Roederer, M.</dc:creator>
<dc:creator>SenGupta, D.</dc:creator>
<dc:creator>Nixon, D.</dc:creator>
<dc:creator>Van Weyenbergh, J.</dc:creator>
<dc:date>2018-11-19</dc:date>
<dc:identifier>doi:10.1101/473363</dc:identifier>
<dc:title><![CDATA[Age but not disease progression defines CD4+ and CD8+ T stem cell memory levels in human retroviral infections: contrasting effects of HTLV-1 and HIV-1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/473447v1?rss=1">
<title>
<![CDATA[
Evaluating Clinical Benefit in I-SPY 2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/473447v1?rss=1</link>
<description><![CDATA[
There are limited methods to analyze both efficacy of a drug and toxicity to a patient in response to treatment. We have developed a metric, the clinical benefit index (CBI) which is quantitative, and a scalable way to integrate patient-reported side effects and efficacy for adaptive randomization. Instead of using efficacy as a singular attribute in decision making for agent selection, we will consider the impact on QoL since many of the newer agents will be less toxic. This will be developed in effort to standardize PRO analysis methods. The clinical benefit index provides an alternative framework for evaluation and would help us to develop a regulatory model for approval of those drugs that have less toxicity by providing a replicable model for measuring toxicity and integrating it with efficacy. Reporting of clinical data can inform the management of toxicity and the integration into a clinical benefit index will help promote regulatory evaluation and preference for less toxic drug/combinations.
]]></description>
<dc:creator>Basu, A.</dc:creator>
<dc:creator>Melisko, M.</dc:creator>
<dc:creator>Esserman, L. J.</dc:creator>
<dc:date>2018-11-19</dc:date>
<dc:identifier>doi:10.1101/473447</dc:identifier>
<dc:title><![CDATA[Evaluating Clinical Benefit in I-SPY 2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/475665v1?rss=1">
<title>
<![CDATA[
Glutamate signaling at cytoneme synapses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/475665v1?rss=1</link>
<description><![CDATA[
We investigated the roles of neuronal synapse components for development of the Drosophila air sac primordium (ASP). The ASP, an epithelial tube, extends specialized signaling filopodia called cytonemes that take up signals such as Dpp from the wing imaginal disc. Dpp signaling in the ASP requires that disc cells express Dpp, Synaptobrevin, Synaptotagmin-1, the glutamate transporter, and a voltage-gated calcium channel, and that ASP cells express the Dpp receptor, Synaptotagmin-4 and the AMPA-type glutamate receptor GluRII. Calcium transients in ASP cytonemes correlate with signaling activity. Calcium transients in the ASP require GluRII, are activated by L-glutamate and by stimulation of an optogenetic ion channel expressed in the wing disc, and are inhibited by EGTA and NASPM. Activation of GluRII is essential but not sufficient for signaling. Cytoneme-mediated signaling is glutamatergic.nnSummaryParacrine signals transfer between Drosophila epithelial cells at glutamatergic synapses.
]]></description>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Kornberg, T. B.</dc:creator>
<dc:date>2018-11-21</dc:date>
<dc:identifier>doi:10.1101/475665</dc:identifier>
<dc:title><![CDATA[Glutamate signaling at cytoneme synapses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/476432v1?rss=1">
<title>
<![CDATA[
Temperature-Jump Solution X-ray Scattering Reveals Distinct Motions in a Dynamic Enzyme 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/476432v1?rss=1</link>
<description><![CDATA[
Correlated motions of proteins and their bound solvent molecules are critical to function, but these features are difficult to resolve using traditional structure determination techniques. Time-resolved methods hold promise for addressing this challenge but have relied on the exploitation of exotic protein photoactivity, and are therefore not generalizable. Temperature-jumps (T-jumps), through thermal excitation of the solvent, have been implemented to study protein dynamics using spectroscopic techniques, but their implementation in X-ray scattering experiments has been limited. Here, we perform T-jump small- and wide-angle X-ray scattering (SAXS/WAXS) measurements on a dynamic enzyme, cyclophilin A (CypA), demonstrating that these experiments are able to capture functional intramolecular protein dynamics. We show that CypA displays rich dynamics following a T-jump, and use the resulting time-resolved signal to assess the kinetics of conformational changes in the enzyme. Two relaxation processes are resolved, which can be characterized by Arrhenius behavior. We also used mutations that have distinct functional effects to disentangle the relationship of the observed relaxation processes. A fast process is related to surface loop motions important for substrate specificity, whereas a slower process is related to motions in the core of the protein that are critical for catalytic turnover. These results demonstrate the power of time-resolved X-ray scattering experiments for characterizing protein and solvent dynamics on the s-ms timescale. We expect the T-jump methodology presented here will be useful for understanding kinetic correlations between local conformational changes of proteins and their bound solvent molecules, which are poorly explained by the results of traditional, static measurements of molecular structure.
]]></description>
<dc:creator>Thompson, M. C.</dc:creator>
<dc:creator>Barad, B. A.</dc:creator>
<dc:creator>Wolff, A. M.</dc:creator>
<dc:creator>Cho, H. S.</dc:creator>
<dc:creator>Schotte, F.</dc:creator>
<dc:creator>Schwarz, D. M. C.</dc:creator>
<dc:creator>Anfinrud, P.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2018-11-21</dc:date>
<dc:identifier>doi:10.1101/476432</dc:identifier>
<dc:title><![CDATA[Temperature-Jump Solution X-ray Scattering Reveals Distinct Motions in a Dynamic Enzyme]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/483099v1?rss=1">
<title>
<![CDATA[
Conformational dynamics of the HIV Vif protein complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/483099v1?rss=1</link>
<description><![CDATA[
HIV-1 viral infectivity factor (Vif) is an intrinsically disordered protein responsible for the ubiquitination of the APOBEC3 antiviral proteins. Vif folds when it binds the Cullin-RING E3 ligase CRL5 and the transcription cofactor CBF-{beta}. A five-protein complex containing the substrate receptor (Vif, CBF-{beta}, Elongin-B, Elongin-C) and Cullin5 (CUL5) has a published crystal structure, but dynamics of this VCBC-CUL5 complex have not been characterized. Here, we use Molecular Dynamics (MD) simulations and NMR to characterize the dynamics of the VCBC complex with and without CUL5 and APOBEC3 bound. Our simulations show that the VCBC complex undergoes global dynamics involving twisting and clamshell opening of the complex, while VCBC-CUL5 maintains a more static conformation, similar to the crystal structure. This observation from MD is supported by methyl-transverse relaxation optimized spectroscopy (methyl-TROSY) NMR data, which indicates that the entire VCBC complex without CUL5 is dynamic on the s-ms timescale. Vif binds APOBEC3 to recruit it to the complex, and methyl-TROSY NMR shows that the VCBC complex is more conformationally restricted when bound to APOBEC3F, consistent with our MD simulations. Vif contains a flexible linker region located at the hinge of the VCBC complex, which changes conformation in conjuction with the global dynamics of the complex. Like other ubiquitin substrate receptors, VCBC can exist alone or in complex with CUL5 in cells. Accordingly, the VCBC complex could be a good target for therapeutics that would inhibit full assembly of the ubiquitination complex by stabilizing an alternate VCBC conformation.
]]></description>
<dc:creator>Ball, K. A.</dc:creator>
<dc:creator>Chan, L. M.</dc:creator>
<dc:creator>Stanley, D. J.</dc:creator>
<dc:creator>Tierney, E.</dc:creator>
<dc:creator>Thapa, S.</dc:creator>
<dc:creator>Ta, H. M.</dc:creator>
<dc:creator>Burton, L. A.</dc:creator>
<dc:creator>Binning, J. M.</dc:creator>
<dc:creator>Jacobsen, M.</dc:creator>
<dc:creator>Gross, J.</dc:creator>
<dc:date>2018-11-29</dc:date>
<dc:identifier>doi:10.1101/483099</dc:identifier>
<dc:title><![CDATA[Conformational dynamics of the HIV Vif protein complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/490938v1?rss=1">
<title>
<![CDATA[
Longitudinally persistent cerebrospinal fluid B-cells resist treatment in multiple sclerosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/490938v1?rss=1</link>
<description><![CDATA[
B-cells are key contributors to chronic autoimmune pathology in multiple sclerosis (MS). Clonally related B-cells exist in the cerebrospinal fluid (CSF), meninges, and central nervous system (CNS) parenchyma of MS patients. Longitudinally stable CSF oligoclonal band (OCB) antibody patterns suggest some local CNS B-cell persistence; however, the longitudinal B-cell dynamics within and between the CSF and blood remain unknown. We sought to address this by performing immunoglobulin heavy chain variable region repertoire sequencing on B-cells from longitudinally collected blood and CSF samples of MS patients (n=10). All patients were untreated at the time of the initial sampling; the majority (n=7) were treated with immune modulating therapies 1.2 (+/-0.3 SD) years later during the second sampling. We found clonal persistence of B-cells in the CSF of five patients; these B-cells were frequently immunoglobulin (Ig) class-switched and CD27+. We identified specific blood B-cell subsets that appear to provide input into CNS repertoires over time. We demonstrate complex patterns of clonal B-cell persistence in CSF and blood, even in patients on high-efficacy immune modulating therapy. Our findings support the concept that peripheral B-cell activation and CNS-compartmentalized immune mechanisms are in part therapy-resistant.
]]></description>
<dc:creator>Greenfield, A. L.</dc:creator>
<dc:creator>Dandekar, R.</dc:creator>
<dc:creator>Ramesh, A.</dc:creator>
<dc:creator>Eggers, E. L.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Laurent, S.</dc:creator>
<dc:creator>Harkin, W.</dc:creator>
<dc:creator>Pierson, N. S.</dc:creator>
<dc:creator>Weber, M. S.</dc:creator>
<dc:creator>Henry, R. G.</dc:creator>
<dc:creator>Bischof, A.</dc:creator>
<dc:creator>Cree, B. A. C.</dc:creator>
<dc:creator>Hauser, S. L.</dc:creator>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:creator>von Büdingen, H.- C.</dc:creator>
<dc:date>2018-12-09</dc:date>
<dc:identifier>doi:10.1101/490938</dc:identifier>
<dc:title><![CDATA[Longitudinally persistent cerebrospinal fluid B-cells resist treatment in multiple sclerosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/498550v1?rss=1">
<title>
<![CDATA[
Beyond SNP Heritability: Polygenicity and Discoverability of Phenotypes Estimated with a Univariate Gaussian Mixture Model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/498550v1?rss=1</link>
<description><![CDATA[
Of signal interest in the genetics of human traits is estimating their polygenicity (the proportion of causally associated single nucleotide polymorphisms (SNPs)) and the discoverability (or effect size variance) of the causal SNPs. Narrow-sense heritability is proportional to the product of these quantities. We present a basic model, using detailed linkage disequilibrium structure from an extensive reference panel, to estimate these quantities from genome-wide association studies (GWAS) summary statistics for SNPs with minor allele frequency >1%. We apply the model to diverse phenotypes and validate the implementation with simulations. We find model polygenicities ranging from [~=] 2 x 10-5 to [~=] 4 x 10-3, with discoverabilities similarly ranging over two orders of magnitude. A power analysis allows us to estimate the proportions of phenotypic variance explained additively by causal SNPs at current sample sizes, and map out sample sizes required to explain larger portions of additive SNP heritability. The model also allows for estimating residual inflation.
]]></description>
<dc:creator>Holland, D.</dc:creator>
<dc:creator>Frei, O.</dc:creator>
<dc:creator>Desikan, R.</dc:creator>
<dc:creator>Fan, C. C.</dc:creator>
<dc:creator>Shadrin, A.</dc:creator>
<dc:creator>Smeland, O.</dc:creator>
<dc:creator>Sundar, V. S.</dc:creator>
<dc:creator>Thompson, P.</dc:creator>
<dc:creator>Andreassen, O. A.</dc:creator>
<dc:creator>Dale, A. M.</dc:creator>
<dc:date>2018-12-17</dc:date>
<dc:identifier>doi:10.1101/498550</dc:identifier>
<dc:title><![CDATA[Beyond SNP Heritability: Polygenicity and Discoverability of Phenotypes Estimated with a Univariate Gaussian Mixture Model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/500462v1?rss=1">
<title>
<![CDATA[
Timed inhibition of CDC7 increases CRISPR-Cas9 mediated templated repair 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/500462v1?rss=1</link>
<description><![CDATA[
Repair of double strand DNA breaks (DSBs) can result in gene disruption or precise gene modification via homology directed repair (HDR) from a templating donor DNA. During genome editing, altering cellular responses to DSBs may be an effective strategy to rebalance editing outcomes towards HDR and away from other repair pathways. To identify factors that regulate HDR from a double-stranded DNA donor (dsDonor), we utilized a pooled screen to define the consequences of thousands of individual gene knockdowns during Cas9-initiated HDR from a double strand plasmid donor. We find that templated dsDonor repair pathways are mostly genetically distinct from single strand donor DNA (ssDonor) repair but share aspects that include dependency upon the Fanconi Anemia (FA) pathway. We also identified several factors whose knockdown increases HDR and thus act as repressors of gene modification. Screening available small molecule inhibitors of these repressors revealed that the cell division cycle 7-related protein kinase (CDC7) inhibitor XL413 increases the efficiency of HDR by 2-3 fold in many contexts, including primary T-cells. XL413 stimulates HDR through cell cycle regulation, inducing an early S-phase cell cycle arrest that, to the best of our knowledge, is uncharacterized for Cas9-induced HDR. We anticipate that XL413 and other such rationally developed inhibitors will be useful tools for boosting the efficiency of gene modification.
]]></description>
<dc:creator>Wienert, B.</dc:creator>
<dc:creator>Feng, S. J.</dc:creator>
<dc:creator>Locke, M.</dc:creator>
<dc:creator>Nguyen, D. N.</dc:creator>
<dc:creator>Wyman, S. K.</dc:creator>
<dc:creator>Kazane, K. R.</dc:creator>
<dc:creator>MARSON, A.</dc:creator>
<dc:creator>Richardson, C. D.</dc:creator>
<dc:creator>Corn, J. E.</dc:creator>
<dc:date>2018-12-19</dc:date>
<dc:identifier>doi:10.1101/500462</dc:identifier>
<dc:title><![CDATA[Timed inhibition of CDC7 increases CRISPR-Cas9 mediated templated repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/502187v1?rss=1">
<title>
<![CDATA[
Exploration of Anti-Yo and Anti-Hu paraneoplastic neurological disorders by PhIP-Seq reveals a highly restricted pattern of antibody epitopes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/502187v1?rss=1</link>
<description><![CDATA[
Paraneoplastic neurological disorders (PNDs) are immune-mediated diseases of the nervous system understood to manifest as part of a misdirected anti-tumor immune response. Identifying PND-associated autoantibodies and their cognate antigens can assist with proper diagnosis and treatment while also enhancing our understanding of tumor-associated immune processes, triggers for autoimmune disease, and the functional significance of onconeuronal proteins. Here, we employed an enhanced version of phage display immunoprecipitation and sequencing (PhIP-Seq) leveraging a library of over 731,000 unique phage clones tiling across the entire human proteome to detect autoantibodies and create high-resolution epitope profiles in serum and CSF samples from patients suffering from two common PNDs, the anti-Yo (n = 36 patients) and anti-Hu syndromes (n = 44 patients). All patient samples positive for anti-Yo antibody by a validated clinical assay yielded polyspecific enrichment of phage presenting peptides from the canonical anti-Yo (CDR2 and CDR2L) antigens, while 38% of anti-Hu patients (17/44) had a serum and/or CSF sample that significantly enriched peptides deriving from the ELAVL family of proteins, the anti-Hu autoantigenic target. The anti-Hu antibodies showed a remarkably convergent antigenic signature across 15/17 patients corresponding to residues surrounding and including the degenerate motif, RLDxLL, shared by ELAVL2, 3 and 4. Lastly, PhIP-Seq identified several known and novel autoantigens in these same patient samples, representing potential biomarkers that could aid in the diagnosis and prognosis of PND and cancer.
]]></description>
<dc:creator>O'Donovan, B. D.</dc:creator>
<dc:creator>Mandel-Brehm, C.</dc:creator>
<dc:creator>Vazquez, S. E.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Parent, A. V.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>Kassimatis, T.</dc:creator>
<dc:creator>Zekeridou, A.</dc:creator>
<dc:creator>Hauser, S. L.</dc:creator>
<dc:creator>Pittock, S. J.</dc:creator>
<dc:creator>Chow, E.</dc:creator>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:date>2018-12-20</dc:date>
<dc:identifier>doi:10.1101/502187</dc:identifier>
<dc:title><![CDATA[Exploration of Anti-Yo and Anti-Hu paraneoplastic neurological disorders by PhIP-Seq reveals a highly restricted pattern of antibody epitopes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/507350v1?rss=1">
<title>
<![CDATA[
Human, Nonhuman Primate, and Bat Cells Are Broadly Susceptible to Tibrovirus Particle Cell Entry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/507350v1?rss=1</link>
<description><![CDATA[
In 2012, the genome of a novel rhabdovirus, Bas-Congo virus, was discovered in the acute-phase serum of a Congolese patient with presumed viral hemorrhagic fever. In the absence of a replicating virus isolate, fulfilling Kochs postulates to determine whether Bas-Congo virus is indeed a human virus and/or pathogen has been impossible. However, experiments with vesiculoviral particles pseudotyped with Bas-Congo glycoprotein suggested that Bas-Congo virus particles can enter cells from multiple animals, including humans. In 2015, genomes of two related viruses, Ekpoma virus 1 and Ekpoma virus 2, were detected in human sera in Nigeria. Isolates could not be obtained. Phylogenetic analyses led to the classification of Bas-Congo virus, Ekpoma virus 1, and Ekpoma virus 2 in the same genus, Tibrovirus, together with five biting midge-borne rhabdoviruses (i.e., Beatrice Hill virus, Bivens Arm virus, Coastal Plains virus, Sweetwater Branch virus, and Tibrogargan virus) not known to infect humans. Using individual recombinant vesiculoviruses expressing the glycoproteins of all eight known tibroviruses and more than 75 cell lines representing different animal species, we demonstrate that the glycoproteins of all tibroviruses can mediate vesiculovirus particle entry into human, bat, nonhuman primate, cotton rat, boa constrictor, and Asian tiger mosquito cells. Using four of five isolated authentic tibroviruses (i.e., Bivens Arm virus, Coastal Plains virus, Sweetwater Branch virus, and Tibrogargan virus), our experiments indicate that many cell types may be partially resistant to tibrovirus replication after virion cell entry. Consequently, experimental data solely obtained from experiments using tibrovirus surrogate systems (e.g., vesiculoviral pseudotypes, recombinant vesiculoviruses) cannot be used to predict whether Bas-Congo virus, or any other tibrovirus, infects humans.
]]></description>
<dc:creator>Cai, Y.</dc:creator>
<dc:creator>Yu, S.</dc:creator>
<dc:creator>Jangra, R.</dc:creator>
<dc:creator>Postnikova, E.</dc:creator>
<dc:creator>Wada, J.</dc:creator>
<dc:creator>Tesh, R.</dc:creator>
<dc:creator>Whelan, S.</dc:creator>
<dc:creator>Lauck, M.</dc:creator>
<dc:creator>Wiley, M.</dc:creator>
<dc:creator>Finch, C.</dc:creator>
<dc:creator>Radoshitzky, S.</dc:creator>
<dc:creator>O'Connor, D.</dc:creator>
<dc:creator>Palacios, G.</dc:creator>
<dc:creator>Chandran, K.</dc:creator>
<dc:creator>Chiu, C.</dc:creator>
<dc:creator>Kuhn, J.</dc:creator>
<dc:date>2018-12-27</dc:date>
<dc:identifier>doi:10.1101/507350</dc:identifier>
<dc:title><![CDATA[Human, Nonhuman Primate, and Bat Cells Are Broadly Susceptible to Tibrovirus Particle Cell Entry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/515288v1?rss=1">
<title>
<![CDATA[
Reversible silencing of endogenous receptors in intact brain tissue using two-photon pharmacology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/515288v1?rss=1</link>
<description><![CDATA[
The physiological activity of proteins is often studied with loss-of-function genetic approaches, but the corresponding phenotypes develop slowly and can be confounding. Photopharmacology allows direct, fast and reversible control of endogenous protein activity, with spatiotemporal resolution set by the illumination method. Here, we combine a photoswitchable allosteric modulator (alloswitch) and two-photon excitation (2PE) using pulsed near-infrared lasers to reversibly silence mGlu5 receptor activity in intact brain tissue. Endogenous receptors can be photoactivated in neurons and astrocytes with pharmacological selectivity and with an axial resolution between 5 and 10 m. Thus, two-photon pharmacology (2PP) using alloswitch allows investigating mGlu5-dependent processes in wild type animals, including synaptic formation and plasticity, and signaling pathways from intracellular organelles.
]]></description>
<dc:creator>Pittolo, S.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Lladoo, A.</dc:creator>
<dc:creator>Tosi, S.</dc:creator>
<dc:creator>Bosch, M.</dc:creator>
<dc:creator>Bardia, L.</dc:creator>
<dc:creator>Gomez-Santacana, X.</dc:creator>
<dc:creator>Llebaria, A.</dc:creator>
<dc:creator>Soriano, E.</dc:creator>
<dc:creator>Colombelli, J.</dc:creator>
<dc:creator>Poskanzer, K. E.</dc:creator>
<dc:creator>Perea, G.</dc:creator>
<dc:creator>Gorostiza, P.</dc:creator>
<dc:date>2019-01-09</dc:date>
<dc:identifier>doi:10.1101/515288</dc:identifier>
<dc:title><![CDATA[Reversible silencing of endogenous receptors in intact brain tissue using two-photon pharmacology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/515569v1?rss=1">
<title>
<![CDATA[
Dynamic Change of Electrostatic Field in TMEM16F Permeation Pathway Shifts Its Ion Selectivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/515569v1?rss=1</link>
<description><![CDATA[
TMEM16F is activated by elevated intracellular Ca2+, and functions both as a small-conductance ion channel permeable to Ca2+ and as a phospholipid scramblase. It is important to hold this positive feedback in check to prevent prolonged Ca2+-overloading in cells. We hypothesize that TMEM16F shifts its ion selectivity so that it is more permeable to Cl- than cations at high intracellular Ca2+ concentration. We tested this hypothesis with the Q559K mutant that shows no current rundown in excised patch with prolonged Ca2+ elevation. Recorded in NaCl-based solution, the channel shifted its ion selectivity from Na+-selective to Cl--selective when intracellular Ca2+ was increased. The ion selectivity switch did not correlate with changes of channel open state. Rather, it was indicative of an alteration of electrostatic field in the permeation pathway. Shifting ion-selectivity synergistically by intracellular divalent ions and membrane potential could work as a built-in mechanism that allows TMEM16F to brake the positive feedback.
]]></description>
<dc:creator>Ye, W.</dc:creator>
<dc:creator>Han, T. W.</dc:creator>
<dc:creator>He, M.</dc:creator>
<dc:creator>Jan, Y. N.</dc:creator>
<dc:creator>Jan, L. Y.</dc:creator>
<dc:date>2019-01-16</dc:date>
<dc:identifier>doi:10.1101/515569</dc:identifier>
<dc:title><![CDATA[Dynamic Change of Electrostatic Field in TMEM16F Permeation Pathway Shifts Its Ion Selectivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/516591v1?rss=1">
<title>
<![CDATA[
Effects of α-tubulin acetylation on microtubule structure and stability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/516591v1?rss=1</link>
<description><![CDATA[
Acetylation of K40 in -tubulin is the sole post-translational modification to mark the luminal surface of microtubules. It is still controversial whether its relationship with microtubule stabilization is correlative or causative. We have obtained high-resolution cryo-electron microscopy reconstructions of pure samples of TAT1-acetylated and SIRT2-deacetylated microtubules to visualize the structural consequences of this modification and reveal its potential for influencing the larger assembly properties of microtubules. We modeled the conformational ensembles of the unmodified and acetylated states by using the experimental cryo-EM density as the structural restraint in molecular dynamics simulations. We found that acetylation alters the conformational landscape of the flexible loop that contains K40. Modification of K40 reduces the disorder of the loop and restricts the states that it samples. We propose that the change in conformational sampling that we describe, at a location very close to the lateral contacts site, is likely to affect microtubule stability and function.
]]></description>
<dc:creator>Eshun-Wilson, L.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Portran, D.</dc:creator>
<dc:creator>Nachury, M.</dc:creator>
<dc:creator>Toso, D.</dc:creator>
<dc:creator>Lohr, T.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Bonomi, M.</dc:creator>
<dc:creator>Fraser, J.</dc:creator>
<dc:creator>Nogales, E.</dc:creator>
<dc:date>2019-01-09</dc:date>
<dc:identifier>doi:10.1101/516591</dc:identifier>
<dc:title><![CDATA[Effects of α-tubulin acetylation on microtubule structure and stability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/523035v1?rss=1">
<title>
<![CDATA[
NWB:N 2.0: An Accessible Data Standard for Neurophysiology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/523035v1?rss=1</link>
<description><![CDATA[
Neurodata Without Borders: Neurophysiology (NWB:N) is a data standard for neurophysiology, providing neuroscientists with a common standard to share, archive, use, and build common analysis tools for neurophysiology data. With NWB:N version 2.0 (NWB:N 2.0) we made significant advances towards creating a usable standard, software ecosystem, and vibrant community for standardizing neurophysiology data. In this manuscript we focus in particular on the NWB:N data standard schema and present advances towards creating an accessible data standard for neurophysiology.
]]></description>
<dc:creator>Ruebel, O.</dc:creator>
<dc:creator>Tritt, A.</dc:creator>
<dc:creator>Dichter, B.</dc:creator>
<dc:creator>Braun, T.</dc:creator>
<dc:creator>Cain, N.</dc:creator>
<dc:creator>Clack, N.</dc:creator>
<dc:creator>Davidson, T. J.</dc:creator>
<dc:creator>Dougherty, M.</dc:creator>
<dc:creator>Fillion-Robin, J.-C.</dc:creator>
<dc:creator>Graddis, N.</dc:creator>
<dc:creator>Grauer, M.</dc:creator>
<dc:creator>Kiggins, J. T.</dc:creator>
<dc:creator>Niu, L.</dc:creator>
<dc:creator>Ozturk, D.</dc:creator>
<dc:creator>Schroeder, W.</dc:creator>
<dc:creator>Soltesz, I.</dc:creator>
<dc:creator>Sommer, F. T.</dc:creator>
<dc:creator>Svoboda, K.</dc:creator>
<dc:creator>Ng, L.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:creator>Bouchard, K.</dc:creator>
<dc:date>2019-01-17</dc:date>
<dc:identifier>doi:10.1101/523035</dc:identifier>
<dc:title><![CDATA[NWB:N 2.0: An Accessible Data Standard for Neurophysiology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/528976v1?rss=1">
<title>
<![CDATA[
Regular cycling between representations of alternatives in the hippocampus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/528976v1?rss=1</link>
<description><![CDATA[
Cognitive faculties such as imagination, planning, and decision-making require the ability to represent alternative scenarios. In animals, split-second decision-making implies that the brain can represent alternatives at a commensurate speed. Yet despite this insight, it has remained unknown whether there exists neural activity that can consistently represent alternatives in <1 s. Here we report that neural activity in the hippocampus, a brain structure vital to cognition, can regularly cycle between representations of alternative locations (bifurcating paths in a maze) at 8 Hz. This cycling dynamic was paced by the internally generated 8 Hz theta rhythm, often occurred in the absence of overt deliberative behavior, and unexpectedly also governed an additional hippocampal representation defined by alternatives (heading direction). These findings implicate a fast, regular, and generalized neural mechanism underlying the representation of competing possibilities.
]]></description>
<dc:creator>Kay, K.</dc:creator>
<dc:creator>Chung, J. E.</dc:creator>
<dc:creator>Sosa, M.</dc:creator>
<dc:creator>Schor, J. S.</dc:creator>
<dc:creator>Karlsson, M. P.</dc:creator>
<dc:creator>Larkin, M. C.</dc:creator>
<dc:creator>Liu, D. F.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:date>2019-01-28</dc:date>
<dc:identifier>doi:10.1101/528976</dc:identifier>
<dc:title><![CDATA[Regular cycling between representations of alternatives in the hippocampus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/529198v1?rss=1">
<title>
<![CDATA[
Microstructure of Ingestive Behaviors in Mus Musculus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/529198v1?rss=1</link>
<description><![CDATA[
We describe a novel quantitative home cage monitoring (HCM) approach for dissecting spontaneous patterns of ingestive and locomotor behaviors into a hierarchically organized series of behavioral facets or endophenotypes. Fine-grained analyses of a large multimodal 16-strain behavioral dataset collected from 169 mice revealed bouts of feeding, drinking and locomotor behaviors occurring within animals Active States. We have automated the detection of these bouts and their discrete properties including bout sizes, rates, durations, and intensities. We have developed a hierarchically organized model of behavioral organization enabling analysis of relationships among Active/Inactive State properties and those of feeding, drinking and locomotor bouts. Robust and analogous patterns of interrelationships among these endophenotypes were found for feeding, drinking behaviors, and these differed markedly from those for locomotor behaviors. For feeding and drinking, patterns of reciprocal relationships were observed for pairs of endophenotypes at multiple hierarchical levels. Moreover, endophenotype variability was highest at lowest hierarchical levels progressively diminished at higher levels, so that variability of gross levels of food and water intake were much less than those of their lower level determinants. By contrast, interrelationships among locomotor endophenotypes differed markedly from those of ingestive behavior. Altogether, these findings raise the possibility that behavioral regulation of food and water intake may make an important contribution to the homeostatic maintenance of energy and volume balance.
]]></description>
<dc:creator>Onnis, G.</dc:creator>
<dc:creator>Layco-Bader, E.</dc:creator>
<dc:creator>Tecott, L.</dc:creator>
<dc:date>2019-02-19</dc:date>
<dc:identifier>doi:10.1101/529198</dc:identifier>
<dc:title><![CDATA[Microstructure of Ingestive Behaviors in Mus Musculus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/533547v1?rss=1">
<title>
<![CDATA[
Multiplex immunofluorescence methods in neurodegenerative diseases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/533547v1?rss=1</link>
<description><![CDATA[
The application of multiplex immunofluorescence to human post-mortem tissue would drive observational studies of selective vulnerability in neurodegenerative proteinopathies. Efficient elution of antibodies is critical for flexibility of antibody combinations and the ability to utilize a sample in multiple rounds of immunostaining. Here, we test two elution strategies for antibodies relevant to the study of selective vulnerability in neurodegenerative diseases in post-mortem human samples from both long-fixed and short-fixed tissue. Both 2-Mercaptoethanol/SDS-based and Urea/SDS/Glycine-based elution strategies work well with the antibodies selected, confirming observations from previous studies with other antibody types.
]]></description>
<dc:creator>Ehrenberg, A. J.</dc:creator>
<dc:creator>Morales, D. O.</dc:creator>
<dc:creator>Tejedor, J. S.</dc:creator>
<dc:creator>Mulder, J.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:date>2019-02-11</dc:date>
<dc:identifier>doi:10.1101/533547</dc:identifier>
<dc:title><![CDATA[Multiplex immunofluorescence methods in neurodegenerative diseases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/534818v1?rss=1">
<title>
<![CDATA[
Population history and gene divergence in Native Mexicans inferred from 76 human exomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/534818v1?rss=1</link>
<description><![CDATA[
Native American genetic variation remains underrepresented in most catalogs of human genome sequencing data. Previous genotyping efforts have revealed that Mexicos indigenous population is highly differentiated and substructured, thus potentially harboring higher proportions of private genetic variants of functional and biomedical relevance. Here we have targeted the coding fraction of the genome and characterized its full site frequency spectrum by sequencing 76 exomes from five indigenous populations across Mexico. Using diffusion approximations, we modeled the demographic history of indigenous populations from Mexico with northern and southern ethnic groups splitting 7.2 kya and subsequently diverging locally 6.5 kya and 5.7 kya, respectively. Selection scans for positive selection revealed BCL2L13 and KBTBD8 genes as potential candidates for adaptive evolution in Raramuris and Triquis, respectively. BCL2L13 is highly expressed in skeletal muscle and could be related to physical endurance, a well-known phenotype of the northern Mexico Raramuri. The KBTBD8 gene has been associated with idiopathic short stature and we found it to be highly differentiated in Triqui, a southern indigenous group from Oaxaca whose height is extremely low compared to other native populations.
]]></description>
<dc:creator>Avila-Arcos, M. C.</dc:creator>
<dc:creator>McManus, K. F.</dc:creator>
<dc:creator>Sandoval, K.</dc:creator>
<dc:creator>Rodriguez-Rodriguez, J. E.</dc:creator>
<dc:creator>Martin, A. R.</dc:creator>
<dc:creator>Luisi, P.</dc:creator>
<dc:creator>Villa-Islas, V.</dc:creator>
<dc:creator>Penaloza-Espinosa, R. I.</dc:creator>
<dc:creator>Eng, C.</dc:creator>
<dc:creator>Huntsman, S.</dc:creator>
<dc:creator>Burchard, E. G.</dc:creator>
<dc:creator>Gignoux, C. R.</dc:creator>
<dc:creator>Bustamante, C. D.</dc:creator>
<dc:creator>Moreno-Estrada, A.</dc:creator>
<dc:date>2019-01-30</dc:date>
<dc:identifier>doi:10.1101/534818</dc:identifier>
<dc:title><![CDATA[Population history and gene divergence in Native Mexicans inferred from 76 human exomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/541656v1?rss=1">
<title>
<![CDATA[
The functional landscape of the human phosphoproteome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/541656v1?rss=1</link>
<description><![CDATA[
Protein phosphorylation is a key post-translational modification regulating protein function in almost all cellular processes. While tens of thousands of phosphorylation sites have been identified in human cells to date, the extent and functional importance of the phosphoproteome remains largely unknown. Here, we have analyzed 6,801 publicly available phospho-enriched mass spectrometry proteomics experiments, creating a state-of-the-art phosphoproteome containing 119,809 human phosphosites. To prioritize functional sites, 59 features indicative of proteomic, structural, regulatory or evolutionary relevance were integrated into a single functional score using machine learning. We demonstrate how this prioritization identifies regulatory phosphosites across different molecular mechanisms and pinpoint genetic susceptibilities at a genomic scale. Several novel regulatory phosphosites were experimentally validated including a role in neuronal differentiation for phosphosites present in the SWI/SNF SMARCC2 complex member. The scored reference phosphoproteome and its annotations identify the most relevant phosphorylations for a given process or disease addressing a major bottleneck in cell signaling studies.
]]></description>
<dc:creator>Ochoa, D.</dc:creator>
<dc:creator>Jarnuczak, A. F.</dc:creator>
<dc:creator>Gehre, M.</dc:creator>
<dc:creator>Soucheray, M.</dc:creator>
<dc:creator>Kleefeldt, A. A.</dc:creator>
<dc:creator>Vieitez, C.</dc:creator>
<dc:creator>Hill, A.</dc:creator>
<dc:creator>Garcia-Alonso, L.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Vizcaino, J. A. A.</dc:creator>
<dc:creator>Noh, K.-M.</dc:creator>
<dc:creator>Beltrao, P.</dc:creator>
<dc:date>2019-02-05</dc:date>
<dc:identifier>doi:10.1101/541656</dc:identifier>
<dc:title><![CDATA[The functional landscape of the human phosphoproteome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/545319v1?rss=1">
<title>
<![CDATA[
Opportunities for improved surveillance and control of infectious diseases from age-specific case data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/545319v1?rss=1</link>
<description><![CDATA[
One of the challenges faced by global disease surveillance efforts is the lack of comparability across systems. Reporting commonly focuses on overall incidence, despite differences in surveillance quality between and within countries. For most immunizing infections, the age-distribution of incident cases provides a more robust picture of trends in transmission. We present a framework to estimate transmission intensity for dengue virus from age-specific incidence data, and apply it to 363 administrative units in Thailand, Colombia, Brazil and Mexico. Our estimates correlate well with those derived from seroprevalence data (the gold-standard), capture the expected spatial heterogeneity in risk, and correlate with known environmental drivers of transmission. We show how this approach could be used to guide the implementation of control strategies such as vaccination. Since age-specific counts are routinely collected by many surveillance systems, they represent a unique opportunity to further our understanding of disease burden and risk for many diseases.
]]></description>
<dc:creator>Rodriguez-Barraquer, I.</dc:creator>
<dc:creator>Salje, H.</dc:creator>
<dc:creator>Cummings, D. A.</dc:creator>
<dc:date>2019-02-08</dc:date>
<dc:identifier>doi:10.1101/545319</dc:identifier>
<dc:title><![CDATA[Opportunities for improved surveillance and control of infectious diseases from age-specific case data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/547356v1?rss=1">
<title>
<![CDATA[
A dataset to explore kinase control of environmental stress responsive transcription 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/547356v1?rss=1</link>
<description><![CDATA[
Cells respond to changes in environmental conditions by activating signal transduction pathways and gene expression programs. Here we present a dataset to explore the relationship between environmental stresses, kinases, and global gene expression in yeast. We subjected 28 drug-sensitive kinase mutants to 10 environmental conditions in the presence of inhibitor and performed mRNA deep sequencing. With these data, we reconstructed canonical stress pathways and identified examples of crosstalk among pathways. The data also implicated numerous kinases in novel environment-specific roles. However, rather than regulating dedicated sets of target genes, individual kinases tuned the magnitude of induction of the environmental stress response (ESR) - a gene expression signature shared across the set of perturbations - in environment-specific ways. This suggests that the ESR integrates inputs from multiple sensory kinases to modulate gene expression and growth control. As an example, we provide experimental evidence that the high osmolarity glycerol pathway is a constitutive negative regulator of protein kinase A, a known inhibitor of the ESR. These results elaborate the central axis of cellular stress response signaling.
]]></description>
<dc:creator>Mace, K.</dc:creator>
<dc:creator>Krakowiak, J.</dc:creator>
<dc:creator>El-Samad, H.</dc:creator>
<dc:creator>Pincus, D.</dc:creator>
<dc:date>2019-02-12</dc:date>
<dc:identifier>doi:10.1101/547356</dc:identifier>
<dc:title><![CDATA[A dataset to explore kinase control of environmental stress responsive transcription]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/548222v1?rss=1">
<title>
<![CDATA[
Cryptic inoviruses are pervasive in bacteria and archaea across Earth's biomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/548222v1?rss=1</link>
<description><![CDATA[
Bacteriophages from the Inoviridae family (inoviruses) are characterized by their unique morphology, genome content, and infection cycle. To date, a relatively small number of inovirus isolates have been extensively studied, either for biotechnological applications such as phage display, or because of their impact on the toxicity of known bacterial pathogens including Vibrio cholerae and Neisseria meningitidis. Here we show that the current 56 members of the Inoviridae family represent a minute fraction of a highly diverse group of inoviruses. Using a new machine learning approach leveraging a combination of marker gene and genome features, we identified 10,295 inovirus-like genomes from microbial genomes and metagenomes. Collectively, these represent six distinct proposed inovirus families infecting nearly all bacterial phyla across virtually every ecosystem. Putative inoviruses were also detected in several archaeal genomes, suggesting that these viruses may have occasionally transferred from bacterial to archaeal hosts. Finally, we identified an expansive diversity of inovirus-encoded toxin-antitoxin and gene expression modulation systems, alongside evidence of both synergistic (CRISPR evasion) and antagonistic (superinfection exclusion) interactions with co-infecting viruses which we experimentally validated in a Pseudomonas model. Capturing this previously obscured component of the global virosphere sparks new avenues for microbial manipulation approaches and innovative biotechnological applications.
]]></description>
<dc:creator>Roux, S.</dc:creator>
<dc:creator>Krupovic, M.</dc:creator>
<dc:creator>Daly, R. A.</dc:creator>
<dc:creator>Borges, A. L.</dc:creator>
<dc:creator>Nayfach, S.</dc:creator>
<dc:creator>Schulz, F.</dc:creator>
<dc:creator>Cheng, J.-F.</dc:creator>
<dc:creator>Ivanova, N. N.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Wrighton, K. C.</dc:creator>
<dc:creator>Woyke, T.</dc:creator>
<dc:creator>Visel, A.</dc:creator>
<dc:creator>Kyrpides, N.</dc:creator>
<dc:creator>Eloe-Fadrosh, E. A.</dc:creator>
<dc:date>2019-02-15</dc:date>
<dc:identifier>doi:10.1101/548222</dc:identifier>
<dc:title><![CDATA[Cryptic inoviruses are pervasive in bacteria and archaea across Earth's biomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/548255v1?rss=1">
<title>
<![CDATA[
Optogenetic control reveals differential promoter interpretation of transcription factor nuclear translocation dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/548255v1?rss=1</link>
<description><![CDATA[
The dynamic translocation of transcription factors (TFs) in and out of the nucleus is thought to encode information, such as the identity of a stimulus. A corollary is the idea that gene promoters can decode different dynamic TF translocation patterns. Testing this TF encoding/promoter decoding hypothesis requires tools that allow direct control of TF dynamics without the pleiotropic effects associated with general perturbations. In this work, we present CLASP (Controllable Light Activated Shuttling and Plasma membrane sequestration), a tool that enables precise, modular, and reversible control of TF localization using a combination of two optimized LOV2 optogenetic constructs. The first sequesters the cargo in the dark at the plasma membrane and releases it upon exposure to blue light, while light exposure of the second reveals a nuclear localization sequence that shuttles the released cargo to the nucleus. CLASP achieves minute-level resolution, reversible translocation of many TF cargos, large dynamic range, and tunable target gene expression. Using CLASP, we investigate the relationship between Crz1, a naturally pulsatile TF, and its cognate promoters. We establish that some Crz1 target genes respond more efficiently to pulsatile TF inputs than to continuous inputs, while others exhibit the opposite behavior. We show using computational modeling that efficient gene expression in response to short pulsing requires fast promoter activation and slow inactivation and that the opposite phenotype can ensue from a multi-stage promoter activation, where a transition in the first stage is thresholded. These data directly demonstrate differential interpretation of TF pulsing dynamics by different genes, and provide plausible models that can achieve these phenotypes.
]]></description>
<dc:creator>Chen, S. Y.</dc:creator>
<dc:creator>Osimiri, L. C.</dc:creator>
<dc:creator>Chevalier, M. W.</dc:creator>
<dc:creator>Bugaj, L. J.</dc:creator>
<dc:creator>Ng, A. H.</dc:creator>
<dc:creator>Stewart-Ornstein, J.</dc:creator>
<dc:creator>Neves, L. T.</dc:creator>
<dc:creator>El-Samad, H.</dc:creator>
<dc:date>2019-02-13</dc:date>
<dc:identifier>doi:10.1101/548255</dc:identifier>
<dc:title><![CDATA[Optogenetic control reveals differential promoter interpretation of transcription factor nuclear translocation dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/549246v1?rss=1">
<title>
<![CDATA[
Dynamics of living cells in a cytomorphological state space 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/549246v1?rss=1</link>
<description><![CDATA[
Cells are non-equilibrium systems that rely on a continuous exchange of matter and energy with the environment to sustain their metabolic needs. The non-equilibrium nature of this system presents considerable challenges to developing a general theory describing its behavior; however, studies have demonstrated that when studied at appropriate spatiotemporal scales, the behavior of ensembles of non-equilibrium systems can resemble that of system at equilibrium. Here we apply this principle to a population of cells within a cytomorphological state space and demonstrate that cellular transition dynamics within this space can be suitably described using equilibrium dynamics formalisms. We use this framework to map the effective energy landscape underlying the cytomorphological state space of a population of mouse embryonic fibroblasts (MEFs) and identify topographical non-uniformity in this space, indicating non-uniform occupation of cytomorphological states within an isogenic population. The introduction of exogenous apoptotic agents altered this energy landscape, inducing formation of additional energy minima that correlated directly with changes in sensitivity to apoptotic induction. The measured application of equilibrium dynamics formalism allows us to accurately capture and these findings suggest that though cells are complex non-equilibrium systems, the application of formalisms derived from equilibrium thermodynamics can provide insight into the basis of non-genetic heterogeneities as well as the relationship between morphological and functional heterogeneity.
]]></description>
<dc:creator>Chang, A. Y.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2019-02-13</dc:date>
<dc:identifier>doi:10.1101/549246</dc:identifier>
<dc:title><![CDATA[Dynamics of living cells in a cytomorphological state space]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/550970v1?rss=1">
<title>
<![CDATA[
Electron cryo-microscopy structures of remodeler-nucleosome intermediates suggest allosteric control through the nucleosome. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/550970v1?rss=1</link>
<description><![CDATA[
The SNF2h remodeler slides nucleosomes most efficiently as a dimer, yet how the two protomers avoid a tug-of-war is unclear. Furthermore, SNF2h couples histone octamer deformation to nucleosome sliding, but the underlying structural basis remains unknown. Here we present cryo-EM structures of SNF2h-nucleosome complexes with ADP-BeFx that capture two reaction intermediates. In one structure, histone residues near the dyad and in the H2A-H2B acidic patch, distal to the active SNF2h protomer, are disordered. The disordered acidic patch is expected to inhibit the second SNF2h promoter, while disorder near the dyad is expected to promote DNA translocation. The other structure doesnt show octamer deformation, but surprisingly shows a 2bp translocation. FRET studies indicate that ADP-BeFx predisposes SNF2h-nucleosome complexes for an elemental translocation step. We propose a model for allosteric control through the nucleosome, where one SNF2h protomer promotes asymmetric octamer deformation to inhibit the second protomer, while stimulating directional DNA translocation.

One sentence summaryCryo-EM structures capture different conformational states of chromatin remodeler-nucleosome complexes.
]]></description>
<dc:creator>Armache, J.-P.</dc:creator>
<dc:creator>Gamarra, N.</dc:creator>
<dc:creator>Johnson, S. L.</dc:creator>
<dc:creator>Leonard, J. D.</dc:creator>
<dc:creator>Wu, S.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:date>2019-02-15</dc:date>
<dc:identifier>doi:10.1101/550970</dc:identifier>
<dc:title><![CDATA[Electron cryo-microscopy structures of remodeler-nucleosome intermediates suggest allosteric control through the nucleosome.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/551978v1?rss=1">
<title>
<![CDATA[
A high-resolution, chromosome-assigned Komodo dragon genome reveals adaptations in the cardiovascular, muscular, and chemosensory systems of monitor lizards 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/551978v1?rss=1</link>
<description><![CDATA[
Monitor lizards are unique among ectothermic reptiles in that they have a high aerobic capacity and distinctive cardiovascular physiology which resembles that of endothermic mammals. We have sequenced the genome of the Komodo dragon (Varanus komodoensis), the largest extant monitor lizard, and present a high resolution de novo chromosome-assigned genome assembly for V. komodoensis, generated with a hybrid approach of long-range sequencing and single molecule physical mapping. Comparing the genome of V. komodoensis with those of related species showed evidence of positive selection in pathways related to muscle energy metabolism, cardiovascular homeostasis, and thrombosis. We also found species-specific expansions of a chemoreceptor gene family related to pheromone and kairomone sensing in V. komodoensis and several other lizard lineages. Together, these evolutionary signatures of adaptation reveal genetic underpinnings of the unique Komodo sensory, cardiovascular, and muscular systems, and suggest that selective pressure altered thrombosis genes to help Komodo dragons evade the anticoagulant effects of their own saliva. As the only sequenced monitor lizard genome, the Komodo dragon genome is an important resource for understanding the biology of this lineage and of reptiles worldwide.
]]></description>
<dc:creator>Lind, A.</dc:creator>
<dc:creator>Lai, Y. Y. Y.</dc:creator>
<dc:creator>Mostovoy, Y.</dc:creator>
<dc:creator>Holloway, A. K.</dc:creator>
<dc:creator>Iannucci, A.</dc:creator>
<dc:creator>Mak, A. C.</dc:creator>
<dc:creator>Fondi, M.</dc:creator>
<dc:creator>Orlandini, V.</dc:creator>
<dc:creator>Eckalbar, W. L.</dc:creator>
<dc:creator>Milan, M.</dc:creator>
<dc:creator>Rovatsos, M.</dc:creator>
<dc:creator>Kichigin, I. G.</dc:creator>
<dc:creator>Makunin, A. I.</dc:creator>
<dc:creator>Trifonov, V.</dc:creator>
<dc:creator>Schijlen, E.</dc:creator>
<dc:creator>Kratochvil, L.</dc:creator>
<dc:creator>Fani, R.</dc:creator>
<dc:creator>Jessop, T. S.</dc:creator>
<dc:creator>Patarnello, T.</dc:creator>
<dc:creator>Hicks, J. W.</dc:creator>
<dc:creator>Ryder, O. A.</dc:creator>
<dc:creator>Mendelson, J. R.</dc:creator>
<dc:creator>Ciofi, C.</dc:creator>
<dc:creator>Kwok, P.-Y. A.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Bruneau, B.</dc:creator>
<dc:date>2019-02-15</dc:date>
<dc:identifier>doi:10.1101/551978</dc:identifier>
<dc:title><![CDATA[A high-resolution, chromosome-assigned Komodo dragon genome reveals adaptations in the cardiovascular, muscular, and chemosensory systems of monitor lizards]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/552075v1?rss=1">
<title>
<![CDATA[
High-throughput single-particle tracking reveals nested membrane nanodomain organization that dictates Ras diffusion and trafficking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/552075v1?rss=1</link>
<description><![CDATA[
Membrane nanodomains have been implicated in Ras signaling, but what these domains are and how they interact with Ras remain obscure. Using high throughput single particle tracking with photoactivated localization microscopy and detailed trajectory analysis, here we show that distinct membrane domains dictate KRas diffusion and trafficking in U2OS cells. KRas exhibits an immobile state in domains [~]70 nm in size, each embedded in a larger domain ([~]200 nm) that confers intermediate mobility, while the rest of the membrane supports fast diffusion. Moreover, KRas is continuously removed from the membrane via the immobile state and replenished to the fast state, likely coupled to internalization and recycling. Importantly, both the diffusion and trafficking properties of KRas remain invariant over a broad range of protein expression levels. Our results reveal how membrane organization dictates KRas diffusion and trafficking and offer insight into how Ras signaling may be regulated through spatial mechanisms.
]]></description>
<dc:creator>Lee, Y.</dc:creator>
<dc:creator>Phelps, C.</dc:creator>
<dc:creator>Huang, T.</dc:creator>
<dc:creator>Mostofian, B.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Stork, P. J. S.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Nan, X.</dc:creator>
<dc:date>2019-02-15</dc:date>
<dc:identifier>doi:10.1101/552075</dc:identifier>
<dc:title><![CDATA[High-throughput single-particle tracking reveals nested membrane nanodomain organization that dictates Ras diffusion and trafficking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/557199v1?rss=1">
<title>
<![CDATA[
Effect of Maxillary Expansion and Protraction on the Oropharyngeal Airway in Individuals with Non-syndromic Cleft Palate with or without Cleft Lip 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/557199v1?rss=1</link>
<description><![CDATA[
IntroductionThe aim of this study was to evaluate three dimensionally the effect of the combined maxillary expansion and protraction treatment on oropharyngeal airway in children with non-syndromic cleft palate with or without cleft lip (CP/L).

MethodsCBCT data of 18 preadolescent individuals (ages, 8.4 {+/-} 1.7 years) with CP/L, who underwent Phase I orthodontic maxillary expansion with protraction, were compared before and after treatment. The average length of treatment was 24.1{+/-} 7.6 months. The airway volume and minimal cross-sectional area (MCA) were determined using 3DMD Vultus imaging software with cross-sectional areas calculated for each 2-mm over the entire length of the airway. A control group of 9 preadolescent individuals (ages, 8.7 {+/-} 2.6 years) with CP/L was used for comparison.

ResultsThere was a statistically significant increase in pharyngeal airway volume after phase I orthodontic treatment in both groups, however, there was no statistically significant change in minimal cross-sectional area in neither study nor control group.

ConclusionThe findings showed that maxillary expansion and protraction did not have a significant effect on increasing oropharyngeal volume and MCA in patients with CP/L.
]]></description>
<dc:creator>Oberoi, S.</dc:creator>
<dc:creator>Alrejaye, N.</dc:creator>
<dc:creator>Gao, J.</dc:creator>
<dc:date>2019-02-21</dc:date>
<dc:identifier>doi:10.1101/557199</dc:identifier>
<dc:title><![CDATA[Effect of Maxillary Expansion and Protraction on the Oropharyngeal Airway in Individuals with Non-syndromic Cleft Palate with or without Cleft Lip]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/560623v1?rss=1">
<title>
<![CDATA[
Brain Aging in Major Depressive Disorder: Results from the ENIGMA Major Depressive Disorder working group 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/560623v1?rss=1</link>
<description><![CDATA[
BackgroundMajor depressive disorder (MDD) is associated with an increased risk of brain atrophy, aging-related diseases, and mortality. We examined potential advanced brain aging in MDD patients, and whether this process is associated with clinical characteristics in a large multi-center international dataset.

MethodsWe performed a mega-analysis by pooling brain measures derived from T1-weighted MRI scans from 29 samples worldwide. Normative brain aging was estimated by predicting chronological age (10-75 years) from 7 subcortical volumes, 34 cortical thickness and 34 surface area, lateral ventricles and total intracranial volume measures separately in 1,147 male and 1,386 female controls from the ENIGMA MDD working group. The learned model parameters were applied to 1,089 male controls and 1,167 depressed males, and 1,326 female controls and 2,044 depressed females to obtain independent unbiased brain-based age predictions. The difference between predicted "brain age" and chronological age was calculated to indicate brain predicted age difference (brain-PAD).

FindingsOn average, MDD patients showed a higher brain-PAD of +0.90 (SE 0.21) years (Cohens d=0.12, 95% CI 0.06-0.17) compared to controls. Relative to controls, first-episode and currently depressed patients showed higher brain-PAD (+1.2 [0.3] years), and the largest effect was observed in those with late-onset depression (+1.7 [0.7] years). In addition, higher brain-PAD was associated with higher self-reported depressive symptomatology (b=0.05, p=0.004).

InterpretationThis highly powered collaborative effort showed subtle patterns of abnormal structural brain aging in MDD. Substantial within-group variance and overlap between groups were observed. Longitudinal studies of MDD and somatic health outcomes are needed to further assess the predictive value of these brain-PAD estimates.

FundingThis work was supported, in part, by NIH grants U54 EB020403 and R01 MH116147.
]]></description>
<dc:creator>Han, L. K.</dc:creator>
<dc:creator>Dinga, R.</dc:creator>
<dc:creator>Hahn, T.</dc:creator>
<dc:creator>Ching, C.</dc:creator>
<dc:creator>Eyler, L.</dc:creator>
<dc:creator>Aftanas, L.</dc:creator>
<dc:creator>Aghajani, M.</dc:creator>
<dc:creator>Aleman, A.</dc:creator>
<dc:creator>Baune, B.</dc:creator>
<dc:creator>Berger, K.</dc:creator>
<dc:creator>Brak, I.</dc:creator>
<dc:creator>Busatto Filho, G.</dc:creator>
<dc:creator>Carballedo, A.</dc:creator>
<dc:creator>Connolly, C.</dc:creator>
<dc:creator>Couvy-Duchesne, B.</dc:creator>
<dc:creator>Cullen, K.</dc:creator>
<dc:creator>Dannlowski, U.</dc:creator>
<dc:creator>Davey, C.</dc:creator>
<dc:creator>Dima, D.</dc:creator>
<dc:creator>Duran, F.</dc:creator>
<dc:creator>Enneking, V.</dc:creator>
<dc:creator>Filimonova, E.</dc:creator>
<dc:creator>Frenzel, S.</dc:creator>
<dc:creator>Frodl, T.</dc:creator>
<dc:creator>Fu, C.</dc:creator>
<dc:creator>Godlewska, B.</dc:creator>
<dc:creator>Gotlib, I.</dc:creator>
<dc:creator>Grabe, H.</dc:creator>
<dc:creator>Groenewold, N.</dc:creator>
<dc:creator>Grotegerd, D.</dc:creator>
<dc:creator>Gruber, O.</dc:creator>
<dc:creator>Hall, G.</dc:creator>
<dc:creator>Harrison, B.</dc:creator>
<dc:creator>Hatton, S.</dc:creator>
<dc:creator>Hermesdorf, M.</dc:creator>
<dc:creator>Hickie, I.</dc:creator>
<dc:creator>Ho, T.</dc:creator>
<dc:creator>Hosten, N.</dc:creator>
<dc:creator>Jansen, A.</dc:creator>
<dc:creator>Kahler, C.</dc:creator>
<dc:creator>Kircher, T.</dc:creator>
<dc:creator>Klimes-Dougan, B.</dc:creator>
<dc:creator>Kramer, B.</dc:creator>
<dc:creator>Krug, A.</dc:creator>
<dc:creator>Lagopoulos, J.</dc:creator>
<dc:creator>Leenings, R.</dc:creator>
<dc:creator>MacMaster</dc:creator>
<dc:date>2019-02-26</dc:date>
<dc:identifier>doi:10.1101/560623</dc:identifier>
<dc:title><![CDATA[Brain Aging in Major Depressive Disorder: Results from the ENIGMA Major Depressive Disorder working group]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/562306v1?rss=1">
<title>
<![CDATA[
Chaperone-Mediated Reflux of Secretory Proteins to the Cytosol During Endoplasmic Reticulum Stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/562306v1?rss=1</link>
<description><![CDATA[
Diverse perturbations to endoplasmic reticulum (ER) functions compromise the proper folding and structural maturation of secretory proteins. To study secretory pathway physiology during such "ER stress", we employed an ER-targeted, redox-responsive, green fluorescent protein--eroGFP--that reports on ambient changes in oxidizing potential. Here we find that diverse ER stress agents cause properly folded, ER-resident eroGFP (and other ER luminal proteins) to "reflux" back to the reducing environment of the cytosol as intact, folded proteins. By utilizing eroGFP in a comprehensive genetic screen in S. cerevisiae, we show that ER protein reflux during ER stress requires specific chaperones and co-chaperones residing in both the ER and the cytosol. Chaperone-mediated ER protein reflux does not require E3 ligase activity, and proceeds even more vigorously when these ER-associated degradation (ERAD) factors are crippled, suggesting that reflux may work in parallel with ERAD. In summary, chaperone-mediated ER-protein reflux may be a conserved protein quality control process that evolved to maintain secretory pathway homeostasis during ER protein-folding stress.nnSIGNIFICANCEApproximately one third of eukaryotic proteins are synthesized on ribosomes attached to the endoplasmic reticulum (ER) membrane. Many of these polypeptides co- or post-translationally translocate into the ER, wherein they fold and mature. An ER quality-control system proofreads these proteins by facilitating their folding and modification, while eliminating misfolded proteins through ER-associated degradation (ERAD). Yet, the fate of many secretory proteins during ER stress is not completely understood. Here, we uncovered an ER-stress induced "protein reflux" system that delivers intact, folded ER luminal proteins back to the cytosol without degrading them. We found that ER protein reflux works in parallel to ERAD and requires distinct ER-resident and cytosolic chaperones and co-chaperones.
]]></description>
<dc:creator>Igbaria, A.</dc:creator>
<dc:creator>Merksamer, P. I.</dc:creator>
<dc:creator>Trusina, A.</dc:creator>
<dc:creator>Tilahun, F.</dc:creator>
<dc:creator>Johnson, J. R.</dc:creator>
<dc:creator>Brandman, O.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:creator>Papa, F. R.</dc:creator>
<dc:date>2019-03-12</dc:date>
<dc:identifier>doi:10.1101/562306</dc:identifier>
<dc:title><![CDATA[Chaperone-Mediated Reflux of Secretory Proteins to the Cytosol During Endoplasmic Reticulum Stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/563445v1?rss=1">
<title>
<![CDATA[
Dynamic chromatin targeting of BRD4 stimulates cardiac fibroblast activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/563445v1?rss=1</link>
<description><![CDATA[
Small molecule inhibitors of the acetyl-histone binding protein BRD4 have been shown to block cardiac fibrosis in pre-clinical models of heart failure (HF). However, the mechanisms by which BRD4 promotes pathological myocardial fibrosis remain unclear. Here, we demonstrate that BRD4 functions as an effector of TGF-{beta} signaling to stimulate conversion of quiescent cardiac fibroblasts into Periostin (Postn)-positive cells that express high levels of extracellular matrix. BRD4 undergoes stimulus-dependent, genome-wide redistribution in cardiac fibroblasts, becoming enriched on a subset of enhancers and super-enhancers, and leading to RNA polymerase II activation and expression of downstream target genes. Employing the SERTA domain-containing protein 4 (Sertad4) locus as a prototype, we demonstrate that dynamic chromatin targeting of BRD4 is controlled, in part, by p38 mitogen-activated protein kinase, and provide evidence of a novel function for Sertad4 in TGF-{beta}-mediated cardiac fibroblast activation. These findings define BRD4 as a central regulator of the pro-fibrotic cell state of cardiac fibroblasts, and establish a signaling circuit for epigenetic reprogramming in HF.
]]></description>
<dc:creator>Stratton, M. S.</dc:creator>
<dc:creator>Bagchi, R. A.</dc:creator>
<dc:creator>Hirsch, R. A.</dc:creator>
<dc:creator>Riching, A. S.</dc:creator>
<dc:creator>Felisbino, M. B.</dc:creator>
<dc:creator>Enyart, B. Y.</dc:creator>
<dc:creator>Koch, K. A.</dc:creator>
<dc:creator>Cavasin, M. A.</dc:creator>
<dc:creator>Alexanian, M.</dc:creator>
<dc:creator>Song, K.</dc:creator>
<dc:creator>Qi, J.</dc:creator>
<dc:creator>Lemieux, M. E.</dc:creator>
<dc:creator>Lam, M. P. Y.</dc:creator>
<dc:creator>Haldar, S. M.</dc:creator>
<dc:creator>Lin, C. Y.</dc:creator>
<dc:creator>McKinsey, T.</dc:creator>
<dc:date>2019-02-28</dc:date>
<dc:identifier>doi:10.1101/563445</dc:identifier>
<dc:title><![CDATA[Dynamic chromatin targeting of BRD4 stimulates cardiac fibroblast activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/563460v1?rss=1">
<title>
<![CDATA[
Theta phase synchrony is sensitive to corollary discharge abnormalities in early illness schizophrenia but not in the clinical high-risk syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/563460v1?rss=1</link>
<description><![CDATA[
BackgroundAcross the animal kingdom, responses in auditory cortex are dampened during vocalizing compared to passive listening, reflecting the action of the corollary discharge mechanism. In humans, it is seen as suppression of the EEG-based N1 event-related potential, with less N1-suppression seen in people with schizophrenia and those at clinical high risk (CHR) for psychosis. Because N1 is an admixture of theta (4-7Hz) power and phase synchrony, we asked which is responsible for N1 effects and if they outperform the sensitivity of N1 to corollary discharge and schizophrenia.

MethodsTheta phase and power values were extracted from EEG data acquired from CHR youth (n=71), early illness schizophrenia patients (ESZ; n=84), and healthy controls (HC; n=103) as they said /ah/ (Talk) and then listened to the sounds played back (Listen). A principal components analysis extracted theta inter-trial coherence (ITC; phase consistency) and event related spectral power, peaking in the N1 latency range.

ResultsTheta ITC-suppression (Cohen d=1.46) was greater than N1-suppression in HC (Cohen d=.63). Both were both reduced in ESZ, but only N1-suppression was reduced in CHR. When deprived of the variance shared with theta-ITC suppression, N1-suppression was no longer sensitive to HC vs. ESZ or HC vs. CHR group differences. Deficits in theta ITC-suppression were correlated with delusions (p=.007) in ESZ. Suppression of theta power was not affected by Group.

ConclusionsTheta ITC-suppression may provide a simpler assay of the corollary discharge mechanism than N1-suppression. Deficits in circuits that generate low frequency oscillations may be an important component of schizophrenia.
]]></description>
<dc:creator>Ford, J. M.</dc:creator>
<dc:creator>Roach, B. J.</dc:creator>
<dc:creator>Loewy, R.</dc:creator>
<dc:creator>Stuart, B.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:date>2019-02-28</dc:date>
<dc:identifier>doi:10.1101/563460</dc:identifier>
<dc:title><![CDATA[Theta phase synchrony is sensitive to corollary discharge abnormalities in early illness schizophrenia but not in the clinical high-risk syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/564310v1?rss=1">
<title>
<![CDATA[
mTORC1 IN THE ORBITOFRONTAL CORTEX PROMOTES HABITUAL ALCOHOL SEEKING 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/564310v1?rss=1</link>
<description><![CDATA[
The mammalian target of rapamycin complex 1 (mTORCl) plays an important role in dendritic translation, synaptic plasticity, and learning and memory. We previously showed that heavy alcohol use activates mTORC1 in the orbitofrontal cortex (OFC) of rodents. Here, we set out to determine the consequences of alcohol-dependent mTORC1 activation in the OFC. We found that although inhibition of mTORC1 in the OFC does not alter rat alcohol intake per se, it attenuates alcohol seeking. We then tested whether mTORC1 in the OFC is required for goal-directed or habitual alcohol seeking. To do so, rats were trained self-administer alcohol under a random ratio (RR) or a random interval (RI) schedule of reinforcement, which biases toward goal-directed or habitual responding, respectively, and tested whether mTORC1 inhibition alters lever presses following alcohol devaluation. We found that pharmacological inhibition of mTORC1 or knockdown of the adaptor protein, Raptor, did not affect goal-directed alcohol responding but restored sensitivity to devaluation in RI-trained rats. In contrast, habitual responding for sucrose was unaltered by mTORC1 inhibition. These data suggest that mTORC1 in the OFC drives alcohol habit. We then elucidate the mechanism by which mTORC1 is activated by alcohol, and found that the recruitment of GluN2B during alcohol withdrawal stimulates mTORC1 in OFC cFos-positive neurons. Finally, we show that inhibition of GluN2B in the OFC attenuates both alcohol seeking and habitual responding for alcohol. Together, our data suggest that alcohol withdrawal promotes an NMDAR-dependent activation of mTORC1 which in turn drives habitual alcohol seeking.
]]></description>
<dc:creator>Morisot, N.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:creator>Phamluong, K.</dc:creator>
<dc:creator>Laguesse, S.</dc:creator>
<dc:creator>Moffat, J.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2019-03-01</dc:date>
<dc:identifier>doi:10.1101/564310</dc:identifier>
<dc:title><![CDATA[mTORC1 IN THE ORBITOFRONTAL CORTEX PROMOTES HABITUAL ALCOHOL SEEKING]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/568139v1?rss=1">
<title>
<![CDATA[
Mapping the human subcortical auditory system using histology, post mortem MRI and in vivo MRI at 7T 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/568139v1?rss=1</link>
<description><![CDATA[
Studying the human subcortical auditory system non-invasively is challenging due to its small, densely packed structures deep within the brain. Additionally, the elaborate three-dimensional (3-D) structure of the system can be difficult to understand based on currently available 2-D schematics and animal models. We addressed these issues using a combination of histological data, post mortem magnetic resonance imaging (MRI), and in vivo MRI at 7 Tesla. We created anatomical atlases based on state-of-the-art human histology (BigBrain) and post mortem MRI (50 m). We measured functional MRI (fMRI) responses to natural sounds and demonstrate that the functional localization of subcortical structures is reliable within individual participants who were scanned in two different experiments. Further, a group functional atlas derived from the functional data locates these structures with a median distance below 2mm. Using diffusion MRI tractography, we revealed structural connectivity maps of the human subcortical auditory pathway both in vivo (1050 m isotropic resolution) and post mortem (200 m isotropic resolution). This work captures current MRI capabilities for investigating the human subcortical auditory system, describes challenges that remain, and contributes novel, openly available data, atlases, and tools for researching the human auditory system.
]]></description>
<dc:creator>Sitek, K. R.</dc:creator>
<dc:creator>Gulban, O. F.</dc:creator>
<dc:creator>Calabrese, E.</dc:creator>
<dc:creator>Johnson, G. A.</dc:creator>
<dc:creator>Ghosh, S. S.</dc:creator>
<dc:creator>de Martino, F.</dc:creator>
<dc:date>2019-03-05</dc:date>
<dc:identifier>doi:10.1101/568139</dc:identifier>
<dc:title><![CDATA[Mapping the human subcortical auditory system using histology, post mortem MRI and in vivo MRI at 7T]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/577924v1?rss=1">
<title>
<![CDATA[
A non-Dicer RNase III and four other novel factors required for RNAi-mediated transposon suppression in the human pathogenic yeast C. neoformans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/577924v1?rss=1</link>
<description><![CDATA[
The human pathogenic yeast Cryptococcus neoformans silences transposable elements using endo-siRNAs and an Argonaute, Ago1. Endo-siRNAs production requires the RNA-dependent RNA polymerase, Rdp1, and two partially redundant Dicer enzymes, Dcr1 and Dcr2, but is independent of histone H3 lysine 9 methylation. We describe here an insertional mutagenesis screen for factors required to suppress the mobilization of the C. neoformans HARBINGER family DNA transposon HAR1. Validation experiments uncovered five novel genes (RDE1-5) required for HAR1 suppression and global production of suppressive endo-siRNAs. Loss of the RDE genes does not impact transcript levels, suggesting the endo-siRNAs do not act by impacting target transcript synthesis or turnover. RDE3 encodes a non-Dicer RNase III related to S. cerevisiae Rnt1, RDE4 encodes a predicted terminal nucleotidyltransferase, while RDE5 has no strongly predicted encoded domains. Affinity purification-mass spectrometry studies reveal that Rde3 and Rde5 are physically associated. RDE1 encodes a G-patch protein homologous to the S. cerevisiae Sqs1/Pfa1, a nucleolar protein that directly activates the essential helicase Prp43 during rRNA biogenesis. Rde1 copurifies Rde2, another novel protein obtained in the screen, as well as Ago1, a homolog of Prp43, and numerous predicted nucleolar proteins. We also describe the isolation of conditional alleles of PRP43, which are defective in RNAi. This work reveals unanticipated requirements for a non-Dicer RNase III and presumptive nucleolar factors for endo-siRNA biogenesis and transposon mobilization suppression in C. neoformans.
]]></description>
<dc:creator>Burke, J. E.</dc:creator>
<dc:creator>Longhurst, A. D.</dc:creator>
<dc:creator>Natarajan, P.</dc:creator>
<dc:creator>Rao, B.</dc:creator>
<dc:creator>Liu, S. J.</dc:creator>
<dc:creator>Sales-Lee, J.</dc:creator>
<dc:creator>Mortensen, Y.</dc:creator>
<dc:creator>Moresco, J.</dc:creator>
<dc:creator>Diedrich, J. K.</dc:creator>
<dc:creator>Yates, J. R.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:date>2019-03-14</dc:date>
<dc:identifier>doi:10.1101/577924</dc:identifier>
<dc:title><![CDATA[A non-Dicer RNase III and four other novel factors required for RNAi-mediated transposon suppression in the human pathogenic yeast C. neoformans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/578542v1?rss=1">
<title>
<![CDATA[
The \"sewing machine\" for minimally invasive neural recording 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/578542v1?rss=1</link>
<description><![CDATA[
We present a system for scalable and customizable recording and stimulation of neural activity. In large animals and humans, the current benchmark for high spatial and temporal resolution neural interfaces are fixed arrays of wire or silicon electrodes inserted into the parenchyma of the brain. However, probes that are large and stiff enough to penetrate the brain have been shown to cause acute and chronic damage and inflammation, which limits their longevity, stability, and yield. One approach to this problem is to separate the requirements of the insertion device, which should to be as stiff as possible, with the implanted device, which should be as small and flexible as possible. Here, we demonstrate the feasibility and scalability of this approach with a system incorporating fine and flexible thin-film polymer probes, a fine and stiff insertion needle, and a robotic insertion machine. Together the system permits rapid and precise implantation of probes, each individually targeted to avoid observable vasculature and to attain diverse anatomical targets. As an initial demonstration of this system, we implanted arrays of electrodes in rat somatosensory cortex, recorded extracellular action potentials from them, and obtained histological images of the tissue response. This approach points the way toward a new generation of scaleable, stable, and safe neural interfaces, both for the basic scientific study of brain function and for clinical applications.
]]></description>
<dc:creator>Hanson, T. L.</dc:creator>
<dc:creator>Diaz-Botia, C. A.</dc:creator>
<dc:creator>Kharazia, V.</dc:creator>
<dc:creator>Maharbiz, M. M.</dc:creator>
<dc:creator>Sabes, P. N.</dc:creator>
<dc:date>2019-03-14</dc:date>
<dc:identifier>doi:10.1101/578542</dc:identifier>
<dc:title><![CDATA[The \"sewing machine\" for minimally invasive neural recording]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/581447v1?rss=1">
<title>
<![CDATA[
Two accessory proteins govern MmpL3 mycolic acid transport in mycobacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/581447v1?rss=1</link>
<description><![CDATA[
Mycolic acids are the signature lipid of mycobacteria and constitute an important physical component of the cell wall, a target of mycobacterial specific antibiotics, and a mediator of M. tuberculosis pathogenesis. Mycolic acids are synthesized in the cytoplasm and are thought to be transported to the cell wall as a trehalose ester by the MmpL3 transporter, an antibiotic target for M. tuberculosis. However, the mechanism by which mycolate synthesis is coupled to transport, and the full MmpL3 transport machinery, is unknown. Here we identify two new components of the MmpL3 transport machinery in mycobacteria. The protein encoded by MSMEG_0736/Rv0383c is essential for growth of M. smegmatis and M. tuberculosis, is anchored to the cytoplasmic membrane, physically interacts with and colocalizes with MmpL3 in growing cells, and is required for trehalose monomycolate transport to the cell wall. In light of these findings we propose Msmeg_0736/Rv0383c be named "TMM transport factor A", TtfA. The protein encoded by MSMEG_5308 also interacts with the MmpL3 complex, but is nonessential for growth or TMM transport. However, MSMEG_5308 accumulates with inhibition of MmpL3 mediated TMM transport and stabilizes the MmpL3/TtfA complex, indicating that it stabilizes the transport system during stress. These studies identify two new components of the mycobacterial mycolate transport machinery, an emerging antibiotic target in M. tuberculosis.
]]></description>
<dc:creator>Fay, A.</dc:creator>
<dc:creator>Czudnochowski, N.</dc:creator>
<dc:creator>Rock, J.</dc:creator>
<dc:creator>Johnson, J.</dc:creator>
<dc:creator>Krogan, N.</dc:creator>
<dc:creator>Rosenberg, O.</dc:creator>
<dc:creator>Glickman, M.</dc:creator>
<dc:date>2019-03-27</dc:date>
<dc:identifier>doi:10.1101/581447</dc:identifier>
<dc:title><![CDATA[Two accessory proteins govern MmpL3 mycolic acid transport in mycobacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/583757v1?rss=1">
<title>
<![CDATA[
Interleukin 17 and senescence regulate the foreign body response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/583757v1?rss=1</link>
<description><![CDATA[
Synthetic biomaterials and medical devices suffer to varying levels from fibrosis via the foreign body response (FBR). To explore mechanistic connections between the immune response and fibrosis from the FBR, we first analyzed fibrotic capsule surrounding human breast implants and found increased numbers of interleukin (IL)17-producing {gamma}{delta}+ T cells and CD4+ TH17 cells as well as senescent cells. Further analysis in a murine model demonstrated an early innate IL17 response to synthetic implants, mediated by innate lymphoid cells and {gamma}{delta}+ T cells, was followed by a chronic adaptive antigen dependent CD4+ TH17 cell response. Mice deficient in IL17 signaling established that IL17 was required for the fibrotic response to materials and the development of p16INK4a senescent cells. Treatment with a senolytic agent reduced IL17 expression and fibrosis. Discovery of a feed-forward loop between the TH17 and senescence response to synthetic materials introduces new targets for therapeutic intervention in the foreign body response.
]]></description>
<dc:creator>Chung, L.</dc:creator>
<dc:creator>Maestas, D.</dc:creator>
<dc:creator>Lebid, A.</dc:creator>
<dc:creator>Mageau, A.</dc:creator>
<dc:creator>Rosson, G. D.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Wolf, M. T.</dc:creator>
<dc:creator>Tam, A.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Vanderzee, I.</dc:creator>
<dc:creator>Andorko, J. I.</dc:creator>
<dc:creator>Narain, R.</dc:creator>
<dc:creator>Sadlter, K.</dc:creator>
<dc:creator>Fan, H.</dc:creator>
<dc:creator>Cihakova, D.</dc:creator>
<dc:creator>Jourdan Le Saux, C.</dc:creator>
<dc:creator>Housseau, F.</dc:creator>
<dc:creator>Pardoll, D.</dc:creator>
<dc:creator>Elisseeff, J.</dc:creator>
<dc:date>2019-03-20</dc:date>
<dc:identifier>doi:10.1101/583757</dc:identifier>
<dc:title><![CDATA[Interleukin 17 and senescence regulate the foreign body response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/585430v1?rss=1">
<title>
<![CDATA[
Whole-genome and RNA sequencing reveal variation and transcriptomic coordination in the developing human prefrontal cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/585430v1?rss=1</link>
<description><![CDATA[
Variation in gene expression underlies neurotypical development, while genomic variants contribute to neuropsychiatric disorders. BrainVar is a unique resource of paired whole-genome sequencing and bulk-tissue RNA-sequencing from the human dorsolateral prefrontal cortex of 176 neurotypical individuals across prenatal and postnatal development, providing the opportunity to assay genomic and transcriptomic variation in tandem. Leveraging this resource, we identified rare premature stop codons with commensurate reduced and allele-specific expression of corresponding genes, and common variants that alter gene expression (expression quantitative trait loci, eQTLs). Categorizing eQTLs by prenatal and postnatal effect, genes affected by temporally-specific eQTLs, compared to constitutive eQTLs, are enriched for haploinsufficiency, protein-protein interactions, and neuropsychiatric disorder risk loci. Expression levels of over 12,000 genes rise or fall in a concerted late-fetal transition, with the transitional genes enriched for cell type specific genes and neuropsychiatric disorder loci, underscoring the importance of cataloguing developmental trajectories in understanding cortical physiology and pathology.nnHighlightsO_LIWhole-genome and RNA-sequencing across human prefrontal cortex development in BrainVarnC_LIO_LIGene-specific developmental trajectories characterize the late-fetal transitionnC_LIO_LIIdentification of constitutive, prenatal-specific, postnatal-specific, and rare eQTLsnC_LIO_LIIntegrated analysis reveals genetic and developmental influences on CNS traits and disordersnC_LI
]]></description>
<dc:creator>Werling, D.</dc:creator>
<dc:creator>Pochareddy, S.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>An, J. Y.</dc:creator>
<dc:creator>Sheppard, B.</dc:creator>
<dc:creator>Peng, M.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Dastmalchi, C.</dc:creator>
<dc:creator>Santperebaro, G.</dc:creator>
<dc:creator>Sousa, A.</dc:creator>
<dc:creator>Tebbenkamp, A.</dc:creator>
<dc:creator>Kaur, N.</dc:creator>
<dc:creator>Gulden, F.</dc:creator>
<dc:creator>Breen, M.</dc:creator>
<dc:creator>Liang, L.</dc:creator>
<dc:creator>Gilson, M.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Dong, S.</dc:creator>
<dc:creator>Klei, L.</dc:creator>
<dc:creator>Cicek, A. E.</dc:creator>
<dc:creator>Buxbaum, J.</dc:creator>
<dc:creator>Adle-Biassette, H.</dc:creator>
<dc:creator>Thomas, J.-L.</dc:creator>
<dc:creator>Aldinger, K. A.</dc:creator>
<dc:creator>O'Day, D. R.</dc:creator>
<dc:creator>Glass, I.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:creator>Talkowski, M.</dc:creator>
<dc:creator>Roeder, K.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Devlin, B.</dc:creator>
<dc:creator>Sanders, S.</dc:creator>
<dc:creator>Sestan, N.</dc:creator>
<dc:date>2019-03-22</dc:date>
<dc:identifier>doi:10.1101/585430</dc:identifier>
<dc:title><![CDATA[Whole-genome and RNA sequencing reveal variation and transcriptomic coordination in the developing human prefrontal cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/585497v1?rss=1">
<title>
<![CDATA[
A NOVEL MECHANISM FOR NF-κB-ACTIVATION VIA IκB-AGGREGATION: IMPLICATIONS FOR HEPATIC MALLORY-DENK-BODY INDUCED INFLAMMATION 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/585497v1?rss=1</link>
<description><![CDATA[
Background & AimsMallory-Denk-bodies (MDBs) are hepatic protein aggregates associated with inflammation both clinically and in MDB-inducing models. Similar protein aggregation in neurodegenerative diseases also triggers inflammation and NF-{kappa}B activation. However, the precise mechanism that links protein aggregation to NF{kappa}B-activation and inflammatory response remains unclear.nnMethodsHerein, we find that treating primary hepatocytes with MDB-inducing agents (N-methylprotoporphyrin, protoporphyrin IX (PPIX), or ZnPPIX) elicited an I{kappa}B-loss with consequent NF-{kappa}B activation. We characterized the underlying mechanism in detail using hepatocytes from various knockout mice and MEF cell lines and multiple approaches including immunoblotting, EMSA, RT-PCR, confocal immunofluorescence microscopy, affinity immunoprecipitation, and protein solubility assays. Additionally, we performed rigorous proteomic analyses to identify the proteins aggregating upon PPIX treatment and/or co-aggregating with I{kappa}B.nnResultsFour known mechanisms of I{kappa}B-loss were probed and excluded. Immunofluorescence analyses of ZnPPIX-treated cells coupled with 8 M urea/CHAPS-extraction revealed that this I{kappa}B-loss was due to its sequestration along with I{kappa}B{beta} into insoluble aggregates. Through proteomic analyses we identified 47 aggregation-prone proteins that co-aggregate with I{kappa}B through direct interaction or proximity. Of these ZnPPIX-aggregation targets, the nucleoporins Nup153 and Nup358/RanBP2 were identified through RNA-interference, as likely mediators of I{kappa}B-nuclear import.nnConclusionWe discovered a novel mechanism of inflammatory NF-{kappa}B activation through I{kappa}B-sequestration into insoluble aggregates along with interacting aggregation-prone proteins. This mechanism may account for the protein aggregate-induced inflammation observed in MDB-associated liver diseases, thereby identifying novel targets for therapeutic intervention. Because of inherent commonalities this MDB cell model is a bona fide protoporphyric model, making these findings equally relevant to the liver inflammation associated with clinical protoporphyria.nnLay SummaryMallory-Denk-bodies (MDBs) are hepatic protein aggregates commonly featured in many liver diseases. MDB-presence is associated with the induction of inflammatory responses both clinically and in all MDB-inducing models. Similar protein aggregation in neurodegenerative diseases is also known to trigger inflammation and NF{kappa}B pathway activation via an as yet to be characterized non-canonical mechanism. Herein using a MDB-inducing cell model, we uncovered a novel mechanism for NF{kappa}B activation via cytosolic I{kappa}B-sequestration into insoluble aggregates. Furthermore, using a proteomic approach, we identified 47 aggregation-prone proteins that interact and co-aggregate with I{kappa}B. This novel mechanism may account for the protein aggregate-induced inflammation observed in liver diseases, thereby identifying novel targets for therapeutic intervention.
]]></description>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Trnka, M. J.</dc:creator>
<dc:creator>Guan, S.</dc:creator>
<dc:creator>Kwon, D.</dc:creator>
<dc:creator>Kim, D.-H.</dc:creator>
<dc:creator>Chen, J.-J.</dc:creator>
<dc:creator>Greer, P. A.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Correia, M. A.</dc:creator>
<dc:date>2019-03-21</dc:date>
<dc:identifier>doi:10.1101/585497</dc:identifier>
<dc:title><![CDATA[A NOVEL MECHANISM FOR NF-κB-ACTIVATION VIA IκB-AGGREGATION: IMPLICATIONS FOR HEPATIC MALLORY-DENK-BODY INDUCED INFLAMMATION]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/597732v1?rss=1">
<title>
<![CDATA[
A novel high-throughput molecular counting method with single base-pair resolution enables accurate single-gene NIPT 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/597732v1?rss=1</link>
<description><![CDATA[
Next-generation DNA sequencing is currently limited by an inability to count the number of input DNA molecules. Molecular counting is particularly needed when accurate quantification is required for diagnostic purposes, such as in single-gene non-invasive prenatal testing (sgNIPT) and liquid biopsy. We developed Quantitative Counting Template (QCT) molecular counting for reconstructing the number of input DNA molecules using sequencing data. We then used QCT molecular counting to develop sgNIPT of sickle cell disease, cystic fibrosis, spinal muscular atrophy, alpha-thalassemia, and beta-thalassemia. Incorporating molecular count information into a statistical model of disease likelihood led to analytical sensitivity and specificity of >98% and >99%, respectively. Validation of sgNIPT was further performed with maternal blood samples collected during pregnancy, and sgNIPT was 100% concordant with newborn follow-up.
]]></description>
<dc:creator>Tsao, D. S.</dc:creator>
<dc:creator>Silas, S.</dc:creator>
<dc:creator>Landry, B. P.</dc:creator>
<dc:creator>Itzep, N.</dc:creator>
<dc:creator>Nguyen, A. B.</dc:creator>
<dc:creator>Kanne, C. K.</dc:creator>
<dc:creator>Sheehan, V. A.</dc:creator>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Shukla, R.</dc:creator>
<dc:creator>Arora, P. N.</dc:creator>
<dc:creator>Atay, O.</dc:creator>
<dc:date>2019-04-03</dc:date>
<dc:identifier>doi:10.1101/597732</dc:identifier>
<dc:title><![CDATA[A novel high-throughput molecular counting method with single base-pair resolution enables accurate single-gene NIPT]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/598805v1?rss=1">
<title>
<![CDATA[
The RNA binding protein DAZL functions as repressor and activator of maternal mRNA translation during oocyte maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/598805v1?rss=1</link>
<description><![CDATA[
Deleted in azoospermia like (DAZL) is an RNA-binding protein playing critical function during gamete development. In fully-grown oocytes, DAZL protein is detected in prophase and levels increase four to five fold during reentry into the meiotic cell cycle. Here, we have investigated the functional significance of this DAZL accumulation in maturing oocytes. Oocyte depletion of DAZL prevents progression to MII. This maturation block is associated with widespread disruption in the pattern of maternal transcripts loading on ribosomes and their translation measured using a RiboTag IP/RNASeq or qPCR strategy. In addition to decreased ribosome loading of a subset of transcripts, we found that DAZL depletion causes also translational activation of distinct subset of mRNAs. DAZL binds to mRNAs whose translation is both repressed and activated during oocyte maturation. Unexpectedly, DAZL depletion also causes increased ribosome loading of a subset of mRNAs in quiescent GV-arrested oocytes. This dual role of repression and activation is recapitulated by using YFP reporters including the 3UTR of DAZL targets. Injection of recombinant DAZL protein in DAZL-depleted oocytes rescues the translation of these targets as well as maturation to MII. Mutagenesis of putative DAZL-binding sites in these candidate mRNAs mimics the effect of DAZL depletion. These findings demonstrate that DAZL regulates translation of maternal mRNAs in mature oocytes, functioning both as translational repressor and activator.
]]></description>
<dc:creator>Yang, C.</dc:creator>
<dc:creator>Rajkovic, G.</dc:creator>
<dc:creator>Daldello, E. M.</dc:creator>
<dc:creator>Luong, X. G.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Conti, M.</dc:creator>
<dc:date>2019-04-04</dc:date>
<dc:identifier>doi:10.1101/598805</dc:identifier>
<dc:title><![CDATA[The RNA binding protein DAZL functions as repressor and activator of maternal mRNA translation during oocyte maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/601013v1?rss=1">
<title>
<![CDATA[
Apoptosis in the fetal testis eliminates developmentally defective germ cell clones 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/601013v1?rss=1</link>
<description><![CDATA[
Many germ cells (GCs) are eliminated during development, long before differentiating to egg or sperm, but it is not clear why. Here, we examined how GC composition in the mouse fetal testis is altered by scheduled apoptosis during sex differentiation. Multicolored-lineage tracing revealed that apoptosis affects clonally-related GCs, suggesting that this fate decision occurs autonomously based on shared intrinsic properties. We identified extensive transcriptional heterogeneity among fetal GCs including an apoptosis-susceptible subpopulation delineated by high Trp53 and deviant differentiation. Alternatively, the GC subpopulation most likely to survive was advanced in differentiation. These results indicate that GC developmental fate is based upon discrete and cell-heritable fitnesses and imply that a dichotomy between sex-differentiation and apoptosis coordinates the removal of developmentally incompetent cells to improve gamete quality. Evidence that GC subpopulations are in different epigenetic states suggests that errors in epigenetic reprogramming form the basis of aberrant differentiation and apoptotic selection.nnOne sentence summaryGerm cells undergo autonomous selection in the fetal testis to promote male differentiation
]]></description>
<dc:creator>Nguyen, D. H.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:date>2019-04-05</dc:date>
<dc:identifier>doi:10.1101/601013</dc:identifier>
<dc:title><![CDATA[Apoptosis in the fetal testis eliminates developmentally defective germ cell clones]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/601229v1?rss=1">
<title>
<![CDATA[
An integrative analysis of GWAS and intermediate molecular trait data reveals common molecular mechanisms supporting genetic similarity between seemingly unrelated complex traits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/601229v1?rss=1</link>
<description><![CDATA[
The rapid accumulation of Genome Wide Association Studies (GWAS) and association studies of intermediate molecular traits provides new opportunities for comparative analysis of the genetic basis of complex human phenotypes. Using a newly developed statistical framework called Sherlock-II that integrates GWAS with eQTL (expression Quantitative Trait Loci) and metabolite-QTL data, we systematically analyzed 445 GWAS datasets, and identified 1371 significant gene-phenotype associations and 308 metabolites-phenotype associations (passing a Q value cutoff of 1/3). This integrative analysis allows us to translate SNP-phenotype associations into functionally informative gene-phenotype association profiles. Genetic similarity analyses based on these profiles clustered phenotypes into sub-trees that reveal both expected and unexpected relationships. We employed a statistical approach to delineate sets of functionally related genes that contribute to the similarity between their association profiles. This approach suggested common molecular mechanisms that connect the phenotypes in a subtree. For example, we found that fasting insulin, fasting glucose, breast cancer, prostate cancer, and lung cancer clustered into a subtree, and identified cyclic AMP/GMP signaling that connects breast cancer and insulin, NAPDH oxidase/ROS generation that connects the three cancers, and apoptosis that connects all five phenotypes. Our approach can be used to assess genetic similarity and suggest mechanistic connections between phenotypes. It has the potential to improve the diagnosis and treatment of a disease by mapping mechanistic insights from one phenotype onto others based on common molecular underpinnings.
]]></description>
<dc:creator>Gu, J.</dc:creator>
<dc:creator>Fuller, C.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2019-04-07</dc:date>
<dc:identifier>doi:10.1101/601229</dc:identifier>
<dc:title><![CDATA[An integrative analysis of GWAS and intermediate molecular trait data reveals common molecular mechanisms supporting genetic similarity between seemingly unrelated complex traits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/604561v1?rss=1">
<title>
<![CDATA[
Rapid discovery of synthetic DNA sequences to rewrite endogenous T cell circuits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/604561v1?rss=1</link>
<description><![CDATA[
Genetically-engineered immune cell therapies have been in development for decades1-3 and recently have proven effective to treat some types of cancer4. CRISPR-based genome editing methods, enabling more flexible and targeted sequence integrations than viral transduction, have the potential to extend the clinical utility of cell therapies5,6. Realization of this potential depends on improved knowledge of how coding and non-coding sites throughout the genome can be modified efficiently and on improved methods to discover novel synthetic DNA sequences that can be introduced at targeted sites to enhance critical immune cell functions. Here, we developed improved guidelines for non-viral genome targeting in human T cells and a pooled discovery platform to identify synthetic genome modifications that enhance therapeutically-relevant cell functions. We demonstrated the breadth of targetable genomic loci by performing large knock-ins at 91 different genomic sites in primary human T cells, and established the power of flexible genome targeting by generating cells with Genetically Engineered Endogenous Proteins (GEEPs) that seamlessly integrate synthetic and endogenous genetic elements to alter signaling input, output, or regulatory control of genes encoding key immune receptors. Motivated by success in introducing synthetic circuits into endogenous sites, we then developed a platform to facilitate discovery of novel multi-gene sequences that reprogram both T cell specificity and function. We knocked in barcoded pools of large DNA sequences encoding polycistronic gene programs. High-throughput pooled screening of targeted knock-ins to the endogenous T cell receptor (TCR) locus revealed a transcriptional regulator and novel protein chimeras that combined with a new TCR specificity to enhance T cell responses in the presence of suppressive conditions in vitro and in vivo. Overall, these pre-clinical studies provide flexible tools to discover complex synthetic gene programs that can be written into targeted genome sites to generate more effective therapeutic cells.
]]></description>
<dc:creator>Roth, T. L.</dc:creator>
<dc:creator>Li, P. J.</dc:creator>
<dc:creator>Nies, J.</dc:creator>
<dc:creator>Yu, R.</dc:creator>
<dc:creator>Nguyen, M. L. T.</dc:creator>
<dc:creator>Lee, Y.</dc:creator>
<dc:creator>Apathy, R.</dc:creator>
<dc:creator>Truong, A.</dc:creator>
<dc:creator>Hiatt, J.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Nguyen, D. N.</dc:creator>
<dc:creator>Goodman, D.</dc:creator>
<dc:creator>Bluestone, J. A.</dc:creator>
<dc:creator>Roybal, K.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2019-04-12</dc:date>
<dc:identifier>doi:10.1101/604561</dc:identifier>
<dc:title><![CDATA[Rapid discovery of synthetic DNA sequences to rewrite endogenous T cell circuits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/607275v1?rss=1">
<title>
<![CDATA[
2’, 3’, 4’-trihydroxychalcone is an Estrogen Receptor Ligand Which Modulates the Activity of 17β-estradiol 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/607275v1?rss=1</link>
<description><![CDATA[
Menopausal hormone therapy (MHT) reduces the risk of osteoporosis, fractures, obesity and diabetes, but long-term MHT increases risk of other diseases. Safe long-term MHT that exploits its benefits and abrogates its adverse effects requires a new approach. Here we demonstrate that 2, 3, 4-trihydroxychalcone (CC7) acts as an estrogen receptor alpha (ER) ligand that may improve the safety profile of MHT. CC7 reprograms the actions of estradiol (E2) to regulate unique genes in bone-derived U2OS cells, with 824/1358 genes not regulated by E2. The proliferative action of E2 on human MCF-7 breast cancer cells and mouse uterus is blocked when combined with CC7. Thermostability and molecular dynamics simulation studies suggest that CC7 binds concurrently with E2 in the ER ligand binding pocket to produce a unique coliganded conformation to modulate ER. Compounds such as CC7 that act as coligands represent a new class of ER reprograming drugs that potentially can be combined with existing estrogens to produce a safer MHT regimen for long-term therapy.
]]></description>
<dc:creator>Herber, C.</dc:creator>
<dc:creator>Quirit, J.</dc:creator>
<dc:creator>Firestone, G.</dc:creator>
<dc:creator>Krois, C.</dc:creator>
<dc:date>2019-04-13</dc:date>
<dc:identifier>doi:10.1101/607275</dc:identifier>
<dc:title><![CDATA[2’, 3’, 4’-trihydroxychalcone is an Estrogen Receptor Ligand Which Modulates the Activity of 17β-estradiol]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/614586v1?rss=1">
<title>
<![CDATA[
Small molecule inhibitors of a human recombination-associated ATPase, RAD54 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/614586v1?rss=1</link>
<description><![CDATA[
Homologous recombination (HR) is a principal support pathway for DNA replication and for recovery from DNA breaks and interstrand crosslinks, making it a rational target for inhibition in cancer therapy. The ATPase RAD54 functions in molecular events that promote DNA sequence-preservation during HR-mediated damage repair, including homology search, DNA strand exchange, and transition to DNA repair synthesis within a displacement loop intermediate. We developed a high-throughput biochemical screen to identify small-molecule inhibitors of human RAD54, using a phosphate detection assay to monitor RAD54 ATPase activity in the presence of double-stranded DNA (dsDNA). After filtering potential DNA intercalators and  frequent hitters, we identified two chemotypes that reproducibly inhibited RAD54 ATPase in vitro. We evaluated these chemotypes for inhibition of RAD54-dsDNA binding and cancer cell survival. A halogenated carbazole/dihydroacridine scaffold inhibited a panel of SWI2/SNF2-related ATPases but not VCP/p97, an unrelated ATPase. Small molecules that interfere with key steps in HR-- such as inhibitors of RAD54--may expose DNA repair-dependent vulnerabilities in cancer cells.
]]></description>
<dc:creator>Ehmsen, K. T.</dc:creator>
<dc:creator>Ang, K. K. H.</dc:creator>
<dc:creator>Wright, W. D.</dc:creator>
<dc:creator>Davies, J. L.</dc:creator>
<dc:creator>Younis, Y.</dc:creator>
<dc:creator>Birman, Y.</dc:creator>
<dc:creator>Bryant, C.</dc:creator>
<dc:creator>Gallardo-Godoy, A.</dc:creator>
<dc:creator>Renslo, A. R.</dc:creator>
<dc:creator>Neitz, R. J.</dc:creator>
<dc:creator>Arkin, M. R.</dc:creator>
<dc:creator>Heyer, W.-D.</dc:creator>
<dc:date>2019-04-19</dc:date>
<dc:identifier>doi:10.1101/614586</dc:identifier>
<dc:title><![CDATA[Small molecule inhibitors of a human recombination-associated ATPase, RAD54]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/618637v1?rss=1">
<title>
<![CDATA[
Modulating pathogenesis with Mobile-CRISPRi 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/618637v1?rss=1</link>
<description><![CDATA[
Pathogens express a set of proteins required for establishing and maintaining an infection, termed virulence life-style genes (VLGs). Due to their outsized importance in pathogenesis, VLG products are attractive targets for the next generation of antimicrobials. However, precise manipulation of VLG expression in the context of infection is technically challenging, limiting our ability to understand the roles of VLGs in pathogenesis and accordingly design effective inhibitors. We previously developed a suite of gene knockdown tools that are transferred by conjugation and stably integrate into pathogen genomes that we call "Mobile-CRISPRi". Here we show the efficacy of Mobile-CRISPRi in controlling VLG expression in a murine infection model. We optimize Mobile-CRISPRi in Pseudomonas aeruginosa for use in a murine model of pneumonia by tuning the expression of CRISPRi components to avoid non-specific toxicity. As a proof of principle, we demonstrate that knockdown of a VLG encoding the type III secretion system (T3SS) activator ExsA blocks effector protein secretion in culture and attenuates virulence in mice. We anticipate that Mobile-CRISPRi will be a valuable tool to probe the function of VLGs across many bacterial species and pathogenesis models.nnImportanceAntibiotic resistance is a growing threat to global health. To optimize the use of our existing antibiotics and identify new targets for future inhibitors, understanding the fundamental drivers of bacterial growth in the context of the host immune response is paramount. Historically these genetic drivers have been difficult to manipulate, as they are requisite for pathogen survival. Here, we provide the first application of Mobile-CRISPRi to study virulence life-style genes in mouse models of lung infection through partial gene perturbation. We envision the use of Mobile-CRISPRi in future pathogenesis models and antibiotic target discovery efforts.
]]></description>
<dc:creator>Rosenberg, O.</dc:creator>
<dc:creator>Qu, J.-X.</dc:creator>
<dc:creator>Prasad, N. K.</dc:creator>
<dc:creator>Peters, J.</dc:creator>
<dc:creator>Looney, M.</dc:creator>
<dc:creator>Yu, M.</dc:creator>
<dc:creator>Lyden, A.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Crawford, E.</dc:creator>
<dc:creator>Silvis, M.</dc:creator>
<dc:date>2019-04-25</dc:date>
<dc:identifier>doi:10.1101/618637</dc:identifier>
<dc:title><![CDATA[Modulating pathogenesis with Mobile-CRISPRi]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/619551v1?rss=1">
<title>
<![CDATA[
Bifunctional Small Molecule Ligands of K-Ras Induce Its Association with Immunophilin Proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/619551v1?rss=1</link>
<description><![CDATA[
Here we report the design, synthesis and characterization of bifunctional chemical ligands that induce the association of Ras with ubiquitously expressed immunophilin proteins such as FKBP12 and cyclophilin A. We show this approach is applicable to two distinct Ras ligand scaffolds, and that both the identity of the immunophilin ligand and the linker chemistry affect compound efficacy in biochemical and cellular contexts. These ligands bind to Ras in an immunophilin-dependent fashion and mediate the formation of tripartite complexes of Ras, immunophilin and the ligand. The recruitment of cyclophilin A to GTP-bound Ras blocks its interaction with B-Raf in biochemical assays. Our study demonstrates the feasibility of ligand-induced association of Ras with intracellular proteins and suggests it as a promising therapeutic strategy for Rasdriven cancers.
]]></description>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:date>2019-04-26</dc:date>
<dc:identifier>doi:10.1101/619551</dc:identifier>
<dc:title><![CDATA[Bifunctional Small Molecule Ligands of K-Ras Induce Its Association with Immunophilin Proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/623041v1?rss=1">
<title>
<![CDATA[
Induction via Functional Protein Stabilization of Hepatic Cytochromes P450 upon gp78/ Autocrine Motility Factor Receptor (AMFR) Ubiquitin E3-ligase Genetic Ablation in Mice: Therapeutic and Toxicological Relevance. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/623041v1?rss=1</link>
<description><![CDATA[
3.The hepatic endoplasmic reticulum (ER)-anchored monotopic proteins, cytochromes P450 (P450s) are enzymes that metabolize endobiotics (physiologically active steroids and fatty acids) as well as xenobiotics including therapeutic/chemotherapeutic drugs, nutrients, carcinogens and toxins. Alterations of hepatic P450 content through synthesis, inactivation or proteolytic turnover influence their metabolic function. P450 proteolytic turnover occurs via ER-associated degradation (ERAD) involving ubiquitin (Ub)-dependent proteasomal degradation (UPD) as a major pathway. UPD critically involves P450 protein ubiquitination by E2/E3 Ub-ligase complexes. We have previously identified the ER-polytopic gp78/AMFR (autocrine motility factor receptor) as a relevant E3 in CYP3A4, CYP3A23 and CYP2E1 UPD. We now document that liver-conditional genetic ablation of gp78/AMFR in mice disrupts P450 ERAD, resulting in significant stabilization of Cyp2a5 and Cyps 2c, in addition to that of Cyps 3a and Cyp2e1. More importantly, we establish that such stabilization is of the functionally active P450 proteins, leading to corresponding significant enhancement of their drug metabolizing capacities. Our findings with clinically relevant therapeutic drugs (nicotine, coumarin, chlorzoxazone, and acetaminophen) and the prodrug (tamoxifen) as P450 substrates, reveal that P450 ERAD disruption could influence therapeutic drug response and/or toxicity, warranting serious consideration as a potential source of clinically significant drug-drug interactions (DDIs). Because gp78/AMFR is not only an E3 Ub-ligase, but also a cell-surface prometastatic oncogene that is upregulated in various malignant cancers, our finding that hepatic gp78/AMFR-knockout can enhance P450-dependent bioactivation of relevant cancer chemotherapeutic prodrugs is of therapeutic relevance and noteworthy in prospective drug design and development.nn4. Significance StatementThe cell surface and ER transmembrane protein gp78/AMFR, a receptor for the prometastatic autocrine motility factor (AMF), as well as an E3 ubiquitin-ligase involved in the ERAD of not only the tumor metastatic suppressor KAI1, but also of hepatic cytochromes P450, is upregulated in various human cancers, enhancing their invasiveness, metastatic potential and poor prognosis. Liver specific gp78/AMFR genetic ablation results in functional protein stabilization of several hepatic P450s and consequently enhanced drug and prodrug metabolism, a feature that could be therapeutically exploited in the bioactivation of chemotherapeutic prodrugs, through design and development of novel short-term gp78/AMFR chemical inhibitors.
]]></description>
<dc:creator>Kwon, D.</dc:creator>
<dc:creator>Kim, S.-m.</dc:creator>
<dc:creator>Jacob, P.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Correia, M. A.</dc:creator>
<dc:date>2019-04-30</dc:date>
<dc:identifier>doi:10.1101/623041</dc:identifier>
<dc:title><![CDATA[Induction via Functional Protein Stabilization of Hepatic Cytochromes P450 upon gp78/ Autocrine Motility Factor Receptor (AMFR) Ubiquitin E3-ligase Genetic Ablation in Mice: Therapeutic and Toxicological Relevance.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/632273v1?rss=1">
<title>
<![CDATA[
Predicting neurodevelopmental outcomes in children with perinatal HIV using a novel machine learning algorithm 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/632273v1?rss=1</link>
<description><![CDATA[
BackgroundA subset of children with perinatal HIV (pHIV) experience long-term neurocognitive symptoms despite treatment with antiretroviral therapy. However, predictors of neurocognitive outcomes remain elusive, particularly for children with pHIV residing in low-to-middle income countries. The present study utilized a novel data analytic approach to identify clinically-relevant predictors of neurocognitive development in children with pHIV.nnMethodsAnalyses were conducted on a large repository of longitudinal data from 285 children with pHIV in Thailand (n=170) and Cambodia (n=115). Participants were designated as neurocognitively resilient (i.e., positive slope; n=143) or at risk (i.e., negative slope; n=142) according to annual performances on the Beery-Buktenica Developmental Test of Visual-Motor Integration over an average of 5.4 years. Gradient-boosted multivariate regression (GBM) with 5-fold cross validation was utilized to identify the optimal combination of demographic, HIV disease, blood markers, and emotional health indices that predicted classification into the two neurocognitive subgroups. Model performance was assessed using Receiver Operator Curves and sensitivity/specificity.nnResultsThe analytic approach distinguished neurocognitive subgroups with high accuracy (93%; sensitivity and specificity each > 90%). Dynamic change indices and interactions between mental health and biological indices emerged as key predictors.nnConclusionMachine learning-based regression defined a unique explanatory model of neurocognitive outcomes among children with pHIV. The predictive algorithm included a combination of HIV, physical health, and mental health indices extracted from readily available clinical measures. Studies are needed to explore the clinical relevance of the data-driven explanatory model, including potential to inform targeted interventions aimed at modifiable risk factors.
]]></description>
<dc:creator>Paul, R.</dc:creator>
<dc:creator>Mellins, C.</dc:creator>
<dc:creator>Robbins, R.</dc:creator>
<dc:creator>Kerr, S.</dc:creator>
<dc:creator>Malee, K.</dc:creator>
<dc:creator>Sophonphan, J.</dc:creator>
<dc:creator>Jahanshad, N.</dc:creator>
<dc:creator>Auripibul, L.</dc:creator>
<dc:creator>Thongpibul, K.</dc:creator>
<dc:creator>Kosalaraksa, P.</dc:creator>
<dc:creator>Kanjanavanit, S.</dc:creator>
<dc:creator>Ngampiyaskul, C.</dc:creator>
<dc:creator>Luesomboon, W.</dc:creator>
<dc:creator>Wongsawat, J.</dc:creator>
<dc:creator>Vonthanak, S.</dc:creator>
<dc:creator>Chettra, K.</dc:creator>
<dc:creator>Suwanlerk, T.</dc:creator>
<dc:creator>Valcour, V.</dc:creator>
<dc:creator>Balla, L.</dc:creator>
<dc:creator>Salminen, L.</dc:creator>
<dc:creator>Preston, R.</dc:creator>
<dc:creator>Garcia, P.</dc:creator>
<dc:creator>Ananworanich, J.</dc:creator>
<dc:creator>Puthanakit, T.</dc:creator>
<dc:date>2019-05-09</dc:date>
<dc:identifier>doi:10.1101/632273</dc:identifier>
<dc:title><![CDATA[Predicting neurodevelopmental outcomes in children with perinatal HIV using a novel machine learning algorithm]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/637041v1?rss=1">
<title>
<![CDATA[
High throughput Characterization of KCNB1 variants Associated with Developmental and Epileptic Encephalopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/637041v1?rss=1</link>
<description><![CDATA[
Pathogenic variants in KCNB1, encoding the voltage-gated potassium channel Kv2.1, are associated with developmental and epileptic encephalopathies (DEE). Previous functional studies on a limited number of KCNB1 variants indicated a range of molecular mechanisms by which variants affect channel function, including loss of voltage sensitivity, loss of ion selectivity, and reduced cell-surface expression. We evaluated a series of 17 KCNB1 variants associated with DEE or neurodevelopmental disorder (NDD) to rapidly ascertain channel dysfunction using high-throughput functional assays. Specifically, we investigated the biophysical properties and cell-surface expression of variant Kv2.1 channels expressed in heterologous cells using high-throughput automated electrophysiology and immunocytochemistry-flow cytometry. Pathogenic variants exhibited diverse functional defects, including altered current density and shifts in the voltage-dependence of activation and/or inactivation, as homotetramers or when co-expressed with wild-type Kv2.1. Quantification of protein expression also identified variants with reduced total Kv2.1 expression or deficient cell-surface expression.nnOur study establishes a platform for rapid screening of functional defects of KCNB1 variants associated with DEE and other NDDs, which will aid in establishing KCNB1 variant pathogenicity and may enable discovery of targeted strategies for therapeutic intervention based on molecular phenotype.
]]></description>
<dc:creator>Kang, S. K.</dc:creator>
<dc:creator>Vanoye, C. G.</dc:creator>
<dc:creator>Misra, S. N.</dc:creator>
<dc:creator>Echevarria, D. M.</dc:creator>
<dc:creator>Calhoun, J. D.</dc:creator>
<dc:creator>O'Connor, J. B.</dc:creator>
<dc:creator>Fabre, K. L.</dc:creator>
<dc:creator>McKnight, D.</dc:creator>
<dc:creator>Demmer, L.</dc:creator>
<dc:creator>Goldenberg, P.</dc:creator>
<dc:creator>Grote, L. E.</dc:creator>
<dc:creator>Thiffault, I.</dc:creator>
<dc:creator>Saunders, C.</dc:creator>
<dc:creator>Strauss, K. A.</dc:creator>
<dc:creator>Torkamani, A.</dc:creator>
<dc:creator>van der Smagt, J.</dc:creator>
<dc:creator>van Gassen, K.</dc:creator>
<dc:creator>Carson, R. P.</dc:creator>
<dc:creator>Diaz, J.</dc:creator>
<dc:creator>Leon, E.</dc:creator>
<dc:creator>Jacher, J. E.</dc:creator>
<dc:creator>Hannibal, M. C.</dc:creator>
<dc:creator>Litwin, J.</dc:creator>
<dc:creator>Friedman, N. R.</dc:creator>
<dc:creator>Schreiber, A.</dc:creator>
<dc:creator>Lynch, B.</dc:creator>
<dc:creator>Poduri, A.</dc:creator>
<dc:creator>Marsh, E. D.</dc:creator>
<dc:creator>Goldberg, E. M.</dc:creator>
<dc:creator>Millichap, J. J.</dc:creator>
<dc:creator>George, A. L.</dc:creator>
<dc:creator>Kearney, J.</dc:creator>
<dc:date>2019-05-14</dc:date>
<dc:identifier>doi:10.1101/637041</dc:identifier>
<dc:title><![CDATA[High throughput Characterization of KCNB1 variants Associated with Developmental and Epileptic Encephalopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/642900v1?rss=1">
<title>
<![CDATA[
Unique Transcription Factor Functions Regulate Epigenetic and Transcriptional Dynamics During Cardiac Reprogramming 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/642900v1?rss=1</link>
<description><![CDATA[
Direct lineage conversion, whereby a somatic cell assumes a new cellular identity, can be driven by ectopic expression of combinations of lineage-enriched transcription factors. To determine the molecular mechanisms by which expression of Gata4, Mef2c, and Tbx5 (GMT) induces direct reprogramming from a cardiac fibroblast toward an induced cardiomyocyte, we performed a comprehensive transcriptomic and epigenomic interrogation of the reprogramming process. Single cell RNA sequencing indicated that a reprogramming trajectory was acquired within 48 hours of GMT introduction, did not require cell division, and was limited mainly by successful expression of GMT. Evaluation of chromatin accessibility by ATAC-seq supported the expression dynamics and revealed widespread chromatin remodeling at early stages of the reprogramming process. Chromatin immunoprecipitation followed by sequencing of each factor alone or in combinations revealed that GMT bind DNA individually and in combination, and that ectopic expression of either Mef2c or Tbx5 is sufficient in some contexts to increase accessibility. We also find evidence for cooperative facilitation and refinement of each factors binding in a combinatorial setting. A random-forest classifier that integrated the observed gene expression dynamics with regions of dynamic chromatin accessibility suggested Tbx5 binding is a primary driver of gene expression changes and revealed additional transcription factor motifs co-segregating with reprogramming factor motifs, suggesting new factors that may be involved in the reprogramming process. These results begin to explain the mechanisms by which transcription factors normally expressed in multiple germ layers can function combinatorially to direct lineage conversion.
]]></description>
<dc:creator>Stone, N. R.</dc:creator>
<dc:creator>Gifford, C. A.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Pratt, K. J. B.</dc:creator>
<dc:creator>Samse-Knapp, K.</dc:creator>
<dc:creator>Mohamed, T. M. A.</dc:creator>
<dc:creator>Radzinsky, E. M.</dc:creator>
<dc:creator>Schricker, A.</dc:creator>
<dc:creator>Yu, P.</dc:creator>
<dc:creator>Ivey, K. N.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:date>2019-05-20</dc:date>
<dc:identifier>doi:10.1101/642900</dc:identifier>
<dc:title><![CDATA[Unique Transcription Factor Functions Regulate Epigenetic and Transcriptional Dynamics During Cardiac Reprogramming]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/643312v1?rss=1">
<title>
<![CDATA[
Huntington phosphorylation governs BDNF homeostasis and improves the phenotype of Mecp2 knockout mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/643312v1?rss=1</link>
<description><![CDATA[
Mutations in the X-linked MECP2 gene are responsible for Rett syndrome (RTT), a severe neurological disorder for which there is no treatment. Several studies have linked the loss of MeCP2 function to alterations of brain-derived neurotrophic factor (BDNF) levels, but non-specific overexpression of BDNF only partially improves the phenotype of Mecp2-deficient mice. We and others have previously shown that huntingtin (HTT) scaffolds molecular motor complexes, transports BDNF-containing vesicles, and is under-expressed in Mecp2 knock-out brains. Here we demonstrate that promoting HTT phosphorylation at Ser421, either by a phospho-mimetic mutation or inhibition of the phosphatase calcineurin, restores endogenous BDNF axonal transport in vitro in the corticostriatal pathway, increases striatal BDNF availability and synaptic connectivity in vivo, and improves the phenotype and the survival of Mecp2 knockout mice--even though treatments were initiated only after the mice had already developed symptoms. Stimulation of endogenous cellular pathways may thus be a promising approach for the treatment of RTT patients.
]]></description>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>Bruyere, j.</dc:creator>
<dc:creator>Panayotis, N.</dc:creator>
<dc:creator>Abada, Y.-S.</dc:creator>
<dc:creator>Saidi, L.</dc:creator>
<dc:creator>Matagne, V.</dc:creator>
<dc:creator>Scaramuzzino, C.</dc:creator>
<dc:creator>Vitet, H.</dc:creator>
<dc:creator>Benoit Delatour, B.</dc:creator>
<dc:creator>Villard, L.</dc:creator>
<dc:creator>Saudou, F.</dc:creator>
<dc:creator>Roux, J.-C.</dc:creator>
<dc:date>2019-05-20</dc:date>
<dc:identifier>doi:10.1101/643312</dc:identifier>
<dc:title><![CDATA[Huntington phosphorylation governs BDNF homeostasis and improves the phenotype of Mecp2 knockout mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/644229v1?rss=1">
<title>
<![CDATA[
Thymically-derived Foxp3+ regulatory T cells are the primary regulators of type 1 diabetes in the non-obese diabetic mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/644229v1?rss=1</link>
<description><![CDATA[
Regulatory T cells (Tregs) are an immunosuppressive population that are identified based on the stable expression of the fate-determining transcription factor forkhead box P3 (Foxp3). Tregs can be divided into distinct subsets based on whether they developed in the thymus (tTregs) or in the periphery (pTregs). Whether there are unique functional roles that distinguish pTregs and tTregs remains largely unclear. To elucidate these functions, efforts have been made to specifically identify and modify individual Treg subsets. Deletion of the conserved non-coding sequence (CNS)1 in the Foxp3 locus leads to selective impairment of pTreg generation without disrupting tTreg generation in the C57BL/6J background. Using CRISPR-Cas9 genome editing technology, we removed the Foxp3 CNS1 region in the non-obese diabetic (NOD) mouse model of spontaneous type 1 diabetes mellitus (T1D) to determine if pTregs contribute to autoimmune regulation. Deletion of CNS1 impaired in vitro induction of Foxp3 in naive NOD CD4+ T cells, but it did not alter Tregs in most lymphoid and non-lymphoid tissues analyzed except for the large intestine lamina propria, where a small but significant decrease in ROR{gamma}t+ Tregs and corresponding increase in Helios+ Tregs was observed in NOD CNS1-/- mice. CNS1 deletion also did not alter the development of T1D or glucose tolerance despite increased pancreatic insulitis in pre-diabetic female NOD CNS1-/- mice. CNS1 Furthermore, the proportions of autoreactive Tregs and conventional T cells (Tconvs) within pancreatic islets were unchanged. These results suggest that pTregs dependent on the Foxp3 CNS1 region are not the dominant regulatory population controlling T1D in the NOD mouse model.
]]></description>
<dc:creator>Holohan, D. R.</dc:creator>
<dc:creator>Van Gool, F.</dc:creator>
<dc:creator>Bluestone, J. A.</dc:creator>
<dc:date>2019-05-20</dc:date>
<dc:identifier>doi:10.1101/644229</dc:identifier>
<dc:title><![CDATA[Thymically-derived Foxp3+ regulatory T cells are the primary regulators of type 1 diabetes in the non-obese diabetic mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/646505v1?rss=1">
<title>
<![CDATA[
Novel Small Molecules Targeting the Intrinsically Disordered Structural Ensemble of α-Synuclein Protect Against Diverse α-Synuclein Mediated Dysfunctions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/646505v1?rss=1</link>
<description><![CDATA[
The over-expression and aggregation of -synuclein (Syn) are linked to the onset and pathology of Parkinsons disease. Native monomeric Syn exists in an intrinsically disordered ensemble of interconverting conformations, which has made its therapeutic targeting by small molecules highly challenging. Nonetheless, here we successfully target the monomeric structural ensemble of Syn and thereby identify novel drug-like small molecules that impact multiple pathogenic processes. Using a surface plasmon resonance high-throughput screen, in which monomeric Syn is incubated with microchips arrayed with tethered compounds, we identified novel Syn interacting drug-like compounds. Because these small molecules could impact a variety of Syn forms present in the ensemble, we tested representative hits for impact on multiple Syn malfunctions in vitro and in cells including aggregation and perturbation of vesicular dynamics. We thereby identified a compound that inhibits Syn misfolding and is neuroprotective, multiple compounds that restore phagocytosis impaired by Syn overexpression, and a compound blocking cellular transmission of Syn. Our studies demonstrate that drug-like small molecules that interact with native Syn can impact a variety of its pathological processes. Thus, targeting the intrinsically disordered ensemble of Syn offers a unique approach to the development of small molecule research tools and therapeutics for Parkinsons disease.
]]></description>
<dc:creator>Toth, G.</dc:creator>
<dc:creator>Neumann, T.</dc:creator>
<dc:creator>Berthet, A.</dc:creator>
<dc:creator>Masliah, E.</dc:creator>
<dc:creator>Spencer, B.</dc:creator>
<dc:creator>Tao, J.</dc:creator>
<dc:creator>Jobling, M.</dc:creator>
<dc:creator>Gardai, S.</dc:creator>
<dc:creator>Bertoncini, C.</dc:creator>
<dc:creator>Cremades, N.</dc:creator>
<dc:creator>Bova, M.</dc:creator>
<dc:creator>Ballaron, S.</dc:creator>
<dc:creator>Chen, X.-H.</dc:creator>
<dc:creator>Mao, W.</dc:creator>
<dc:creator>Nguyen, P.</dc:creator>
<dc:creator>Tabios, M.</dc:creator>
<dc:creator>Tambe, M.</dc:creator>
<dc:creator>Rochet, J.-C.</dc:creator>
<dc:creator>Junker, H.-D.</dc:creator>
<dc:creator>Schwizer, D.</dc:creator>
<dc:creator>Sekul, R.</dc:creator>
<dc:creator>Ott, I.</dc:creator>
<dc:creator>Anderson, J.</dc:creator>
<dc:creator>Szoke, B.</dc:creator>
<dc:creator>Hoffman, W.</dc:creator>
<dc:creator>Christodoulou, J.</dc:creator>
<dc:creator>Yednock, T.</dc:creator>
<dc:creator>Agard, D.</dc:creator>
<dc:creator>Schenk, D.</dc:creator>
<dc:creator>McConlogue, L.</dc:creator>
<dc:date>2019-05-24</dc:date>
<dc:identifier>doi:10.1101/646505</dc:identifier>
<dc:title><![CDATA[Novel Small Molecules Targeting the Intrinsically Disordered Structural Ensemble of α-Synuclein Protect Against Diverse α-Synuclein Mediated Dysfunctions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/648238v1?rss=1">
<title>
<![CDATA[
Unraveling the functional role of the orphan solute carrier, SLC22A24 in the transport of steroid conjugates through metabolomic and genome-wide association studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/648238v1?rss=1</link>
<description><![CDATA[
Variation in sex hormone levels has wide implications for health and disease. The genes encoding the proteins involved in steroid disposition represent key determinants of interindividual variation in steroid levels and ultimately, their effects. Beginning with metabolomic data from genome-wide association studies (GWAS), we observed that genetic variants in the orphan transporter, SLC22A24 were significantly associated with levels of androsterone glucuronide and etiocholanolone glucuronide (sentinel SNPs p-value <1x10-30). In cells over-expressing human or various mammalian orthologs of SLC22A24, we showed that steroid conjugates and bile acids were substrates of the transporter. Phylogenetic, genomic, and transcriptomic analyses suggested that SLC22A24 has a specialized role in the kidney and appears to function in the reabsorption of organic anions, and in particular, anionic steroids. Phenome-wide analysis showed that functional variants of SLC22A24 are associated with human disease such as cardiovascular diseases and acne, which have been linked to dysregulated steroid metabolism. Collectively, these functional genomic studies reveal a previously uncharacterized protein involved in steroid homeostasis, opening up new possibilities for SLC22A24 as a pharmacological target for regulating steroid levels.nnAuthor SummarySteroid hormones, ranging from sex steroids such as testosterone to glucocorticoids play key roles in human health and disease. Accordingly, the identification of the genes and proteins involved in their synthesis, disposition and elimination has been the subject of numerous genetic studies. We have been intrigued by recent studies demonstrating that genetic variants in or near a gene encoding SLC22A24 are strongly associated with steroid levels. SLC22A24 is an orphan transporter with no known ligands and no known biological functions. In this study, we use cellular and computational methods to show that SLC22A24 transports steroid conjugates, bile acids and other dicarboxylic acids. Based on the direction of association of a common stop codon in SLC22A24 with lower levels of steroids, our studies suggest that the transporter functions to reabsorb steroid conjugates in the kidney, a surprising finding, given that conjugation pathways generally function to polar molecules that are readily eliminated by the kidney. The absence of the transporter gene in many species and its presence in higher order primates suggest that SLC22A24 plays a specialized role in steroid homeostasis. Overall, our studies indicate that SLC22A24 functions in the reabsorption of conjugated steroids in the kidney.
]]></description>
<dc:creator>Yee, S. W.</dc:creator>
<dc:creator>Stecula, A.</dc:creator>
<dc:creator>Chien, H.-C.</dc:creator>
<dc:creator>Zou, L.</dc:creator>
<dc:creator>Feofanova, E. V.</dc:creator>
<dc:creator>van Borselen, M.</dc:creator>
<dc:creator>Cheung, K. W. K.</dc:creator>
<dc:creator>Yousri, N. A.</dc:creator>
<dc:creator>Suhre, K.</dc:creator>
<dc:creator>Kinchen, J. M.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Irannejad, R.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Giacomini, K. M.</dc:creator>
<dc:date>2019-05-23</dc:date>
<dc:identifier>doi:10.1101/648238</dc:identifier>
<dc:title><![CDATA[Unraveling the functional role of the orphan solute carrier, SLC22A24 in the transport of steroid conjugates through metabolomic and genome-wide association studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/649020v1?rss=1">
<title>
<![CDATA[
cfrB,cfrC, and a potential new cfr-like gene in Clostridium difficile strains recovered across Latin America 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/649020v1?rss=1</link>
<description><![CDATA[
Cfr is a radical S-adenosyl-L-methionine (SAM) enzyme that confers cross-resistance to all antibiotics targeting the large ribosomal subunit through hypermethylation of nucleotide A2503 of 23S rRNA. Of the four known cfr genes known to date, cfr(B) and cfr(C) have been sporadically found in C. difficile, yet functional characterization of cfr(C) is still lacking. We identified genes for putative Cfr-like enzymes among clinical C. difficile strains from Mexico, Honduras, Costa Rica, and Chile. To confirm their identity and activity, we obtained minimum inhibitory concentrations for ribosome-targeting antibiotics, annotated whole genome sequences, and performed a functional characterization of Cfr(C). The seven representative isolates analyzed displayed different levels of resistance to PhLOPSA antibiotics in the absence of the ribosome protection factor OptrA, and mutations in genes for 23S rRNAs or the ribosomal proteins L3 and L4. cfr(B) was detected in four isolates as part of a Tn6218-like transposon or an un-described mobile genetic element. In turn, cfr(C) was found integrated into an ICE-element. One isolate harbored a putative cfr-like gene that shows only 51-58% of sequence identity to Cfr and known Cfr-like enzymes. Moreover, our in vitro assays confirmed that Cfr(C) methylates E. coli and C. difficile 23S rRNA fragments. These results indicate selection of cfr-like genes in C. difficile from Latin America, suggest that the diversity of cfr-like resistance genes is larger than anticipated, and provide the first assessment of the methylation activity of Cfr(C).
]]></description>
<dc:creator>Stojkovic, V.</dc:creator>
<dc:creator>Ulate, M. F.</dc:creator>
<dc:creator>Hidalgo-Villeda, F.</dc:creator>
<dc:creator>Aguilar, E.</dc:creator>
<dc:creator>Monge-Cascante, C.</dc:creator>
<dc:creator>Pizarro-Guajardo, M.</dc:creator>
<dc:creator>Tsai, K.</dc:creator>
<dc:creator>Tzoc, E.</dc:creator>
<dc:creator>Camorlinga-Ponce, M.</dc:creator>
<dc:creator>Paredes-Sabja, D.</dc:creator>
<dc:creator>Quesada-Gomez, C.</dc:creator>
<dc:creator>Fujimori, D.</dc:creator>
<dc:creator>Rodriguez, C.</dc:creator>
<dc:date>2019-05-24</dc:date>
<dc:identifier>doi:10.1101/649020</dc:identifier>
<dc:title><![CDATA[cfrB,cfrC, and a potential new cfr-like gene in Clostridium difficile strains recovered across Latin America]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/653337v1?rss=1">
<title>
<![CDATA[
Investigating Transfusion-Related Sepsis using Culture-Independent Metagenomic Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/653337v1?rss=1</link>
<description><![CDATA[
BackgroundTransfusion-related sepsis remains an important hospital infection control challenge. Investigating septic transfusion events is often restricted by the limitations of bacterial culture in terms of time requirements and low yield in the setting of prior antibiotic administration.nnMethodsIn three Gram-negative septic transfusion cases, we performed mNGS of direct clinical blood specimens in addition to standard culture-based approaches utilized for infection control investigations. Pathogen detection leveraged IDSeq, a new open-access microbial bioinformatics portal. Phylogenetic analysis was performed to assess microbial genetic relatedness and understand transmission events.nnResultsmNGS of direct clinical blood specimens afforded precision detection of pathogens responsible for each case of transfusion-related sepsis, and enabled discovery of a novel Acinetobacter species in a platelet product that had become contaminated despite photochemical pathogen reduction. In each case, longitudinal assessment of pathogen burden elucidated the temporal sequence of events associated with each transfusion-transmitted infection. We found that informative data could be obtained from culture-independent mNGS of residual platelet products and leftover blood specimens that were either unsuitable or unavailable for culture, or that failed to grow due to prior antibiotic administration. We additionally developed methods to enhance accuracy for detecting transfusion-associated pathogens sharing taxonomic similarity to contaminants commonly found in mNGS library preparations.nnConclusionsCulture-independent mNGS of blood products afforded rapid and precise assessment of pathogen identity, abundance and genetic relatedness. Together, these challenging cases demonstrated the potential for metagenomics to advance existing methods for investigating transfusion-transmitted infections.
]]></description>
<dc:creator>Crawford, E. D.</dc:creator>
<dc:creator>Kamm, J.</dc:creator>
<dc:creator>Miller, S.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Caldera, S.</dc:creator>
<dc:creator>Lyden, A. D.</dc:creator>
<dc:creator>Yokoe, D.</dc:creator>
<dc:creator>Nichols, A.</dc:creator>
<dc:creator>Ramirez-Avila, L.</dc:creator>
<dc:creator>Tran, N. K.</dc:creator>
<dc:creator>Barnhard, S.</dc:creator>
<dc:creator>Conner, P. M.</dc:creator>
<dc:creator>Nambiar, A.</dc:creator>
<dc:creator>Zinter, M. S.</dc:creator>
<dc:creator>Moayeri, M.</dc:creator>
<dc:creator>Hayakawa Serpa, P.</dc:creator>
<dc:creator>Prince, B. C.</dc:creator>
<dc:creator>Quan, J.</dc:creator>
<dc:creator>Sit, R.</dc:creator>
<dc:creator>Tan, M.</dc:creator>
<dc:creator>Phelps, M.</dc:creator>
<dc:creator>DeRisi, J. S.</dc:creator>
<dc:creator>Tato, C. M.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:date>2019-05-30</dc:date>
<dc:identifier>doi:10.1101/653337</dc:identifier>
<dc:title><![CDATA[Investigating Transfusion-Related Sepsis using Culture-Independent Metagenomic Sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/666230v1?rss=1">
<title>
<![CDATA[
Serological and metagenomic interrogation of cerebrospinal fluid implicates enteroviruses in pediatric acute flaccid myelitis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/666230v1?rss=1</link>
<description><![CDATA[
BackgroundSince 2014, the United States has experienced a biennial spike in pediatric acute flaccid myelitis (AFM). Epidemiologic evidence suggests non-polio enteroviruses (EVs) are a potential etiology, yet EV RNA is rarely detected in cerebrospinal fluid (CSF) and only inconsistently identified from the respiratory tract, serum, or stool.nnMethodsWe interrogated CSF from children with AFM (n=42) and pediatric controls with other neurologic diseases (OND) (n=58). Samples were incubated with T7 bacteriophage expressing 481,966 sixty-two amino acid peptides with a fourteen amino acid overlap tiled across all known vertebrate virus and arbovirus genomes, an adaption of the VirScan method. Antibody-bound phage were deep sequenced to quantify enriched peptides with normalized counts expressed as reads per hundred thousand (rpK). EV antibody findings were confirmed with ELISA using whole viral protein 1 (VP1) from contemporary enterovirus (EV) A71 and D68 strains. Separately, metagenomic next-generation sequencing (mNGS) of CSF RNA, both unbiased and with targeted enrichment for EVs, was performed.nnResultsThe most significantly enriched viral family by VirScan of CSF in AFM versus OND controls was Picornaviridae (mean rpK 11,266 versus mean rpK 950, p-adjusted < 0.001, Wilcoxon signed-rank test with Bonferroni adjustment). Enriched Picornaviridae peptides belonged almost entirely to the genus Enterovirus. The mean EV VP1 ELISA signal in AFM (mean OD 0.51) was significantly higher than OND controls (mean OD 0.08, p-value < 0.001, Mann-Whitney test). mNGS did not detect additional enterovirus RNA in CSF.nnConclusionDespite the rare detection of EV RNA in the CNS of patients with AFM, a pan-viral serologic assay identified high levels of CSF EV antibodies in AFM CSF compared to CSF from OND controls. These results provide further evidence for a causal role of non-polio enteroviruses in AFM.
]]></description>
<dc:creator>Schubert, R. D.</dc:creator>
<dc:creator>Hawes, I.</dc:creator>
<dc:creator>Ramachandran, P. S.</dc:creator>
<dc:creator>Ramesh, A.</dc:creator>
<dc:creator>Crawford, E. D.</dc:creator>
<dc:creator>Pak, J. E.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Cheung, C. K.</dc:creator>
<dc:creator>O'Donovan, B. D.</dc:creator>
<dc:creator>Tato, C. M.</dc:creator>
<dc:creator>Lyden, A.</dc:creator>
<dc:creator>Tan, M.</dc:creator>
<dc:creator>Sit, R.</dc:creator>
<dc:creator>Sowa, G.</dc:creator>
<dc:creator>Sample, H. A.</dc:creator>
<dc:creator>Zorn, K. C.</dc:creator>
<dc:creator>Banerji, D.</dc:creator>
<dc:creator>Khan, L. M.</dc:creator>
<dc:creator>Bove, R.</dc:creator>
<dc:creator>Hauser, S. L.</dc:creator>
<dc:creator>Gelfand, A. A.</dc:creator>
<dc:creator>Johnson-Kerner, B.</dc:creator>
<dc:creator>Nash, K.</dc:creator>
<dc:creator>Krishnamoorthy, K. S.</dc:creator>
<dc:creator>Chitnis, T.</dc:creator>
<dc:creator>Ding, J. Z.</dc:creator>
<dc:creator>McMillan, H. J.</dc:creator>
<dc:creator>Chiu, C. Y.</dc:creator>
<dc:creator>Briggs, B.</dc:creator>
<dc:creator>Glaser, C. A.</dc:creator>
<dc:creator>Yen, C.</dc:creator>
<dc:creator>Chu, V.</dc:creator>
<dc:creator>Wadford, D. A.</dc:creator>
<dc:creator>Dominguez, S. R.</dc:creator>
<dc:creator>Ng, T. F. F.</dc:creator>
<dc:creator>Marine, R. L.</dc:creator>
<dc:creator>Lopez, A. S.</dc:creator>
<dc:creator>Nix, W. A.</dc:creator>
<dc:creator>Soldatos, A.</dc:creator>
<dc:creator>Gorman, M.</dc:creator>
<dc:creator>Benson, L.</dc:creator>
<dc:creator>Messacar, K.</dc:creator>
<dc:creator>Konopka-Anstadt, J. L.</dc:creator>
<dc:creator>Ob</dc:creator>
<dc:date>2019-06-10</dc:date>
<dc:identifier>doi:10.1101/666230</dc:identifier>
<dc:title><![CDATA[Serological and metagenomic interrogation of cerebrospinal fluid implicates enteroviruses in pediatric acute flaccid myelitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/674291v1?rss=1">
<title>
<![CDATA[
Comparison of Rosetta flexible-backbone computational protein design methods on binding interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/674291v1?rss=1</link>
<description><![CDATA[
Computational design of binding sites in proteins remains difficult, in part due to limitations in our current ability to sample backbone conformations that enable precise and accurate geometric positioning of side chains during sequence design. Here we present a benchmark framework for comparison between flexible-backbone design methods applied to binding interactions. We quantify the ability of different flexible backbone design methods in the widely used protein design software Rosetta to recapitulate observed protein sequence profiles assumed to represent functional protein/protein and protein/small molecule binding interactions. The CoupledMoves method, which combines backbone flexibility and sequence exploration into a single acceptance step during the sampling trajectory, better recapitulates observed sequence profiles than the BackrubEnsemble and FastDesign methods, which separate backbone flexibility and sequence design into separate acceptance steps during the sampling trajectory. Flexible-backbone design with the CoupledMoves method is a powerful strategy for reducing sequence space to generate targeted libraries for experimental screening and selection.
]]></description>
<dc:creator>Loshbaugh, A. L.</dc:creator>
<dc:creator>Kortemme, T.</dc:creator>
<dc:date>2019-06-17</dc:date>
<dc:identifier>doi:10.1101/674291</dc:identifier>
<dc:title><![CDATA[Comparison of Rosetta flexible-backbone computational protein design methods on binding interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/675645v1?rss=1">
<title>
<![CDATA[
BOLD Decoding of Individual Pain Anticipation Biases During Uncertainty 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/675645v1?rss=1</link>
<description><![CDATA[
A prominent model of pain as a predictive cue posits that anticipation shapes pain transmission and ultimately pain experience. Consistent with this model, the neural mechanisms underlying pain anticipation have the power to modulate pain experience thus understanding pain predictions, particularly during uncertainty, may allow us to ascertain measures indicative of intrinsic anticipation biases. Understanding such biases moves way to precision pain management, as it can guide the individualized treatment. To examine individual pain anticipation biases, we applied machine-learning-based neural decoding to functional magnetic resonance imaging (fMRI) data acquired during a pain-anticipation paradigm to identify individualized neural activation patterns differentiating two certain anticipatory conditions, which we then used to decode that individuals uncertain anticipatory condition. We showed that neural patterns representative of the individualized response during certain anticipatory conditions were differentiable with high accuracy and, across individuals, most commonly involved neural activation patterns within anterior short gyrus of the insula and the nucleus accumbens. Using unsupervised clustering of individualized decodings of anticipatory responses during uncertain conditions, we identified three distinct response profiles representing subjects who, in uncertain situations, consistently anticipated high-pain (i.e., negative bias), subjects who consistently anticipated low-pain (i.e., positive bias), and subjects whose decoded anticipation responses were depended on the intensity of the preceding pain stimulus. The individualized decoded pain anticipation biases during uncertainty were independent of existence or type of diagnosed psychopathology, were stable over one year timespan and were related to underlying insula anatomy. Our results suggest that anticipation behaviors may be intrinsic, stable, and specific to each individual. Understanding individual differences in the neurobiology of pain anticipation has the potential to greatly improve the clinical pain management.
]]></description>
<dc:creator>Kadlec, M.</dc:creator>
<dc:creator>Tosun, D.</dc:creator>
<dc:creator>Strigo, I.</dc:creator>
<dc:date>2019-06-20</dc:date>
<dc:identifier>doi:10.1101/675645</dc:identifier>
<dc:title><![CDATA[BOLD Decoding of Individual Pain Anticipation Biases During Uncertainty]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/677682v1?rss=1">
<title>
<![CDATA[
Regulation of community functional composition across taxonomic variation by resource-consumer dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/677682v1?rss=1</link>
<description><![CDATA[
High-throughput sequencing techniques such as metagenomic and metatranscriptomic technologies allow cataloging of functional characteristics of microbial community members as well as their taxonomic identity. Such studies have found that a communitys composition in terms of ecologically relevant functional traits or guilds can be conserved more strictly across varying settings than taxonomic composition is. I use a standard ecological resource-consumer model to examine the dynamics of traits relevant to resource consumption, and analyze determinants of functional composition. This model demonstrates that interaction with essential resources can regulate the community-wide abundance of ecologically relevant traits, keeping them at consistent levels despite large changes in the abundances of the species housing those traits in response to changes in the environment, and across variation between communities in species composition. Functional composition is shown to be able to track differences in environmental conditions faithfully across differences in community composition. Mathematical conditions on consumers vital rates and functional responses sufficient to produce conservation of functional community structure across taxonomic differences are presented.
]]></description>
<dc:creator>Worden, L.</dc:creator>
<dc:date>2019-06-21</dc:date>
<dc:identifier>doi:10.1101/677682</dc:identifier>
<dc:title><![CDATA[Regulation of community functional composition across taxonomic variation by resource-consumer dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/678060v1?rss=1">
<title>
<![CDATA[
Mapping gene regulatory networks of primary CD4+ T cells using single-cell genomics and genome engineering 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/678060v1?rss=1</link>
<description><![CDATA[
Gene regulatory programs controlling the activation and polarization of CD4+ T cells are incompletely mapped and the interindividual variability in these programs remain unknown. We sequenced the transcriptomes of ~160k CD4+ T cells from 9 donors following pooled CRISPR perturbation targeting 140 regulators. We identified 134 regulators that affect T cell functionalization, including IRF2 as a positive regulator of Th2 polarization. Leveraging correlation patterns between cells, we mapped 194 pairs of interacting regulators, including known (e.g. BATF and JUN) and novel interactions (e.g. ETS1 and STAT6). Finally, we identified 80 natural genetic variants with effects on gene expression, 48 of which are modified by a perturbation. In CD4+ T cells, CRISPR perturbations can influence in vitro polarization and modify the effects of trans and cis regulatory elements on gene expression.
]]></description>
<dc:creator>Gate, R. E.</dc:creator>
<dc:creator>Kim, M. C.</dc:creator>
<dc:creator>Lu, A.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Subramaniam, M.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:date>2019-06-21</dc:date>
<dc:identifier>doi:10.1101/678060</dc:identifier>
<dc:title><![CDATA[Mapping gene regulatory networks of primary CD4+ T cells using single-cell genomics and genome engineering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/681809v1?rss=1">
<title>
<![CDATA[
Adipocyte JAK2 mediates aging-associated metabolic liver disease and progression to hepatocellular carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/681809v1?rss=1</link>
<description><![CDATA[
Non-alcoholic fatty liver disease (NAFLD) and steatohepatitis (NASH) are liver manifestations of the metabolic syndrome and can progress to hepatocellular carcinoma (HCC). Loss of Growth Hormone (GH) signaling is reported to predispose to NAFLD and NASH through direct actions on the liver. Here, we report that aged mice lacking hepatocyte Jak2 (JAK2L), an obligate transducer of Growth Hormone (GH) signaling, spontaneously develop the full spectrum of phenotypes found in patients with metabolic liver disease, beginning with insulin resistance and lipodystrophy and manifesting as NAFLD, NASH and even HCC, independent of dietary intervention. Remarkably, insulin resistance, metabolic liver disease, and carcinogenesis are prevented in JAK2L mice via concomitant deletion of adipocyte Jak2 (JAK2LA). Further, we demonstrate that GH increases hepatic lipid burden but does so indirectly via signaling through adipocyte JAK2. Collectively, these data establish adipocytes as the mediator of GH-induced metabolic liver disease and carcinogenesis. In addition, we report a new spontaneous model of NAFLD, NASH, and HCC that recapitulates the natural sequelae of human insulin resistance-associated disease progression. The work presented here suggests a attention be paid towards inhibition of adipocyte GH signaling as a therapeutic target of metabolic liver disease.
]]></description>
<dc:creator>Corbit, K. J.</dc:creator>
<dc:creator>Wilson, C. G.</dc:creator>
<dc:creator>Lowe, D.</dc:creator>
<dc:creator>Tran, J. L.</dc:creator>
<dc:creator>Vera, N. B.</dc:creator>
<dc:creator>Clasquin, M.</dc:creator>
<dc:creator>Mattis, A. N.</dc:creator>
<dc:creator>Weiss, E. J.</dc:creator>
<dc:date>2019-06-24</dc:date>
<dc:identifier>doi:10.1101/681809</dc:identifier>
<dc:title><![CDATA[Adipocyte JAK2 mediates aging-associated metabolic liver disease and progression to hepatocellular carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/683672v1?rss=1">
<title>
<![CDATA[
Using a bar-coded AAV capsid library to select for novel clinically relevant gene therapy vectors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/683672v1?rss=1</link>
<description><![CDATA[
ABSTRACTWhile gene transfer using recombinant adeno-associated viral (rAAV) vectors have shown success in some clinical trials, there remain many tissues that are not well transduced. Because of the recent success in reprogramming islet derived cells into functional {beta}-cells in animal models, we constructed two highly complex barcoded replication competent capsid shuffled libraries and selected for high transducing variants on primary human islets. We describe a chimeric capsid (AAV-KP1) that penetrated and transduced primary human islet cells and human embryonic stem cell derived {beta}-cells with up to 10-fold higher efficiency compared to previously studied best in class AAV vectors. Remarkably, this chimeric capsid was also able to transduce both mouse and human hepatocytes at very high levels in a humanized-chimeric mouse model, thus providing a versatile vector which has the potential to be used in both preclinical testing and human clinical trials for both liver-based diseases and diabetes.
]]></description>
<dc:creator>Pekrun, K.</dc:creator>
<dc:creator>De Alencastro, G.</dc:creator>
<dc:creator>Luo, Q.-J.</dc:creator>
<dc:creator>Kim, Y.</dc:creator>
<dc:creator>Nygaard, S.</dc:creator>
<dc:creator>Galivo, F.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Song, R.</dc:creator>
<dc:creator>Tiffany, M.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Hebron, M.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Kay, M. A.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/683672</dc:identifier>
<dc:title><![CDATA[Using a bar-coded AAV capsid library to select for novel clinically relevant gene therapy vectors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/685933v1?rss=1">
<title>
<![CDATA[
What is that molecular machine really doing? Automated exploration of alternative transporter mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/685933v1?rss=1</link>
<description><![CDATA[
Motivated by growing evidence for pathway heterogeneity and alternative functions of molecular machines, we demonstrate a computational approach for investigating two questions: (1) Are there multiple mechanisms (state-space pathways) by which a machine can perform a given function, such as cotransport across a membrane? (2) How can additional functionality, such as proofreading/error-correction, be built into machine function using standard biochemical processes? Answers to these questions will aid both the understanding of molecular-scale cell biology and the design of synthetic machines. Focusing on transport in this initial study, we sample a variety of mechanisms by employing Metropolis Markov chain Monte Carlo. Trial moves adjust transition rates among an automatically generated set of conformational and binding states while maintaining fidelity to thermodynamic principles and a user-supplied fitness/functionality goal. Each accepted move generates a new model. The simulations yield both single and mixed reaction pathways for cotransport in a simple environment with a single substrate along with a driving ion. In a "competitive" environment including an additional decoy substrate, several qualitatively distinct reaction pathways are found which are capable of extremely high discrimination coupled to a leak of the driving ion, akin to proofreading. The array of functional models would be difficult to find by intuition alone in the complex state-spaces of interest.

Author summaryMolecular machines, which operate on the nanoscale, are proteins/complexes that perform remarkable tasks such as the selective absorption of nutrients into the cell by transporters. These complex machines are often described using a fairly simple set of states and transitions that may not account for the stochasticity and heterogeneity generally expected at the nanoscale at body temperature. New tools are needed to study the full array of possibilities. This study presents a novel in silico method to systematically generate testable molecular-machine kinetic models and explore alternative mechanisms, applied first to membrane transport proteins. Our initial results suggest these transport machines may contain mechanisms which  detoxify the cell of an unwanted toxin, as well as significantly discriminate against the import of the toxin. This novel approach should aid the experimental study of key physiological processes such as renal glucose re-absorption, rational drug design, and potentially the development of synthetic machines.
]]></description>
<dc:creator>George, A.</dc:creator>
<dc:creator>Bisignano, P.</dc:creator>
<dc:creator>Rosenberg, J.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2019-06-28</dc:date>
<dc:identifier>doi:10.1101/685933</dc:identifier>
<dc:title><![CDATA[What is that molecular machine really doing? Automated exploration of alternative transporter mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/690297v1?rss=1">
<title>
<![CDATA[
Tumor cell phenotype and heterogeneity differences in IDH1 mutant vs wild-type gliomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/690297v1?rss=1</link>
<description><![CDATA[
Glioma is recognized to be a highly heterogeneous CNS malignancy, whose diverse cellular composition and cellular interactions have not been well characterized. To gain new clinical- and biological-insights into the genetically-bifurcated IDH1 mutant (mt) vs wildtype (wt) forms of glioma, we integrated multiplexed immunofluorescence single cell data for 43 protein markers across cancer hallmarks, in addition to cell spatial metrics, genomic sequencing and magnetic resonance imaging (MRI) quantitative features. Molecular and spatial heterogeneity scores for angiogenesis and cell invasion differ between IDHmt and wt gliomas irrespective of prior treatment and tumor grade; these differences also persisted in the MR imaging features of peritumoral edema and contrast enhancement volumes. Longer overall survival for IDH1mt glioma patients may reflect generalized altered cellular, molecular, spatial heterogeneity which manifest in discernable radiological manifestations.
]]></description>
<dc:creator>Berens, M.</dc:creator>
<dc:creator>Sood, A.</dc:creator>
<dc:creator>Barnholtz-Sloan, J.</dc:creator>
<dc:creator>Graf, J. F.</dc:creator>
<dc:creator>Cho, S.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Kiefer, J.</dc:creator>
<dc:creator>Byron, S. A.</dc:creator>
<dc:creator>Halperin, R. F.</dc:creator>
<dc:creator>Nasser, S.</dc:creator>
<dc:creator>Adkins, J.</dc:creator>
<dc:creator>Cuyugan, L.</dc:creator>
<dc:creator>Devine, K.</dc:creator>
<dc:creator>Ostrom, Q.</dc:creator>
<dc:creator>Couce, M.</dc:creator>
<dc:creator>Wolansky, L.</dc:creator>
<dc:creator>McDonough, E.</dc:creator>
<dc:creator>Schyberg, S.</dc:creator>
<dc:creator>Dinn, S.</dc:creator>
<dc:creator>Sloan, A. E.</dc:creator>
<dc:creator>Prados, M.</dc:creator>
<dc:creator>Phillips, J. J.</dc:creator>
<dc:creator>Nelson, S. J.</dc:creator>
<dc:creator>Liang, W. S.</dc:creator>
<dc:creator>Rusu, M.</dc:creator>
<dc:creator>Zavodszky, M.</dc:creator>
<dc:creator>Ginty, F.</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/690297</dc:identifier>
<dc:title><![CDATA[Tumor cell phenotype and heterogeneity differences in IDH1 mutant vs wild-type gliomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/696724v1?rss=1">
<title>
<![CDATA[
Mapping Vector Field of Single Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/696724v1?rss=1</link>
<description><![CDATA[
Single-cell RNA-seq, together with RNA velocity and metabolic labeling, reveals cellular states and transitions at unprecedented resolution. Fully exploiting these data, however, requires dynamical models capable of predicting cell fate and unveiling the governing regulatory mechanisms. Here, we introduce dynamo, an analytical framework that reconciles intrinsic splicing and labeling kinetics to estimate absolute RNA velocities, reconstructs velocity vector fields that predict future cell fates, and finally employs differential geometry analyses to elucidate the underlying regulatory networks. We applied dynamo to a wide range of disparate biological processes including prediction of future states of differentiating hematopoietic stem cell lineages, deconvolution of glucocorticoid responses from orthogonal cell-cycle progression, characterization of regulatory networks driving zebrafish pigmentation, and identification of possible routes of resistance to SARS-CoV-2 infection. Our work thus represents an important step in going from qualitative, metaphorical conceptualizations of differentiation, as exemplified by Waddingtons epigenetic landscape, to quantitative and predictive theories.
]]></description>
<dc:creator>Qiu, X.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Yang, D.</dc:creator>
<dc:creator>Hosseinzadeh, S.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Yuan, R.</dc:creator>
<dc:creator>Xu, S.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Replogle, J.</dc:creator>
<dc:creator>Darmanis, S.</dc:creator>
<dc:creator>Xing, J.</dc:creator>
<dc:creator>Weissman, J.</dc:creator>
<dc:date>2019-07-09</dc:date>
<dc:identifier>doi:10.1101/696724</dc:identifier>
<dc:title><![CDATA[Mapping Vector Field of Single Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/699215v1?rss=1">
<title>
<![CDATA[
HBMITool: a user-friendly software for labeling Human Brain Microscopy Images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/699215v1?rss=1</link>
<description><![CDATA[
One of the most popular tools for quantifying protein expression is Immunofluorescence (IF). Although IF is widely applied in drug discovery research and assessing disease mechanisms, it has great room for improvement on the task of analyzing human postmortem brain samples. IF analysis of postmortem human tissue relies mostly on manual interaction, which is often error-prone and leading to low inter and intra-observer reproducibility. The high level of autofluorescence caused by accumulation of lipofuscin pigment during aging impedes systematic analyses of human postmortem brain samples. A method for automating cell counting and classification in IF microscopy of human postmortem brains was proposed before, which speeds up the quantification task while improving reproducibility. To correct for misclassified cells by the algorithm, we created HBFMTool, a software package that ease the process of editing the result produced by cell detection/classification algorithm.
]]></description>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Alegro, M.</dc:creator>
<dc:date>2019-07-11</dc:date>
<dc:identifier>doi:10.1101/699215</dc:identifier>
<dc:title><![CDATA[HBMITool: a user-friendly software for labeling Human Brain Microscopy Images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/701698v1?rss=1">
<title>
<![CDATA[
Multiple Measures Reveal The Value of Both Race And Geographic Ancestry For Self-Identification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/701698v1?rss=1</link>
<description><![CDATA[
There is long-standing tension regarding whether and how to use race or geographic ancestry in biomedical research. We examined multiple self-reported measures of race and ancestry from a cohort of over 100,000 U.S. residents alongside genetic data. We found that these measures are often non-overlapping, and that no single self-reported measure alone provides a better fit to genetic ancestry than a combination including both race and geographic ancestry. We also found that patterns of reporting for race and ancestry appear to be influenced by participation in direct-to-consumer genetic ancestry testing. Our results demonstrate that there is a place for the language of both race and geographic ancestry as we seek to empower individuals to fully describe their family history in research and medicine.nnOne Sentence SummarySelf-identification in the United States according to both racial and geographic terms best reflects genetic ancestry in individuals.
]]></description>
<dc:creator>Damotte, V.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Lin, C.</dc:creator>
<dc:creator>Williams, E.</dc:creator>
<dc:creator>Louzoun, Y.</dc:creator>
<dc:creator>Madbouly, A.</dc:creator>
<dc:creator>Kotlarz, R.</dc:creator>
<dc:creator>McDaniel, M.</dc:creator>
<dc:creator>Norman, P. J.</dc:creator>
<dc:creator>Lizee, A.</dc:creator>
<dc:creator>Myres, N. M.</dc:creator>
<dc:creator>Ball, C. A.</dc:creator>
<dc:creator>Chahine, K. G.</dc:creator>
<dc:creator>Byrnes, J.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Maiers, M.</dc:creator>
<dc:creator>Hollenbach, J. A.</dc:creator>
<dc:date>2019-07-13</dc:date>
<dc:identifier>doi:10.1101/701698</dc:identifier>
<dc:title><![CDATA[Multiple Measures Reveal The Value of Both Race And Geographic Ancestry For Self-Identification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/704619v1?rss=1">
<title>
<![CDATA[
Aberrant cell segregation in craniofacial primordia and the emergence of facial dysmorphology in craniofrontonasal syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/704619v1?rss=1</link>
<description><![CDATA[
Craniofrontonasal syndrome (CFNS) is a rare X-linked disorder characterized by craniofacial, skeletal, and neurological anomalies and caused by mutations in EFNB1. Heterozygous females are more severely affected by CFNS than hemizygous male patients, a phenomenon called cellular interference that is correlated with cell segregation resulting from EPHRIN-B1 mosaicism. Efnb1 heterozygous mutant mice also exhibit more severe phenotypes than Efnb1 hemizygous males as well as cell segregation, but how craniofacial dysmorphology arises from cell segregation is unknown and CFNS etiology therefore remains poorly understood. Here, we couple geometric morphometric techniques with temporal and spatial interrogation of embryonic cell segregation in mouse models to elucidate mechanisms underlying CFNS pathogenesis. By generating ephrin-B1 mosaicism at different developmental timepoints and in specific cell populations, we find that ephrin-B1 regulates cell segregation independently in early neural development and later in craniofacial development, correlating with the emergence of quantitative differences in face shape. Whereas specific craniofacial shape changes are qualitatively similar in Efnb1 heterozygous and hemizygous mutant embryos, heterozygous embryos are quantitatively more severely affected, indicating that Efnb1 mosaicism exacerbates loss of function phenotypes rather than having a neomorphic effect. Notably, tissue-specific disruption of Efnb1 throughout neural development does not appear to contribute to CFNS dysmorphology, but its disruption within neural crest cell-derived mesenchyme results in phenotypes very similar to widespread loss. Ephrin-B1 can bind and signal with EphB1, EphB2, and EphB3 receptor tyrosine kinases, but the signaling partner(s) relevant to CFNS are unknown. Geometric morphometric analysis of an allelic series of Ephb1; Ephb2; Ephb3 mutant embryos indicates that EphB2 and EphB3 are key receptors mediating Efnb1 hemizygous-like phenotypes, but the complete loss of EphB1-3 does not recapitulate CFNS-like Efnb1 heterozygous severity. Finally, by generating Efnb1+/-; Ephb1; Ephb2; Ephb3 quadruple knockout mice, we determine how modulating cumulative receptor activity influences cell segregation in craniofacial development and find that while EphB2 and EphB3 play an important role in craniofacial cell segregation, EphB1 is more important for cell segregation in the brain; surprisingly, complete loss of EphB1-EphB3 does not completely abrogate cell segregation. Together, these data advance our understanding of the morphogenetic etiology and signaling interactions underlying CFNS dysmorphology.nnAuthor SummaryCraniofacial anomalies are extremely common, accounting for one third of all birth defects, but even when the responsible genes are known, it often remains to be determined exactly how development has gone wrong. Craniofrontonasal syndrome (CFNS), which affects multiple aspects of craniofacial development, is a particularly mysterious disorder because it is X-linked, but affects females more severely than males, the opposite situation of most X-linked diseases. The responsible gene has been identified as EFNB1, which encodes the EPHRIN-B1 signaling molecule that regulates cellular position. Why EFNB1+/- heterozygous females exhibit severe stereotypical CFNS phenotypes is not well understood, but it is related to the fact that X chromosome inactivation generates mosaicism for EPHRIN-B1. Using mice harboring mutations in the Efnb1 gene in different embryonic tissues, and in receptor genes Ephb1-3, together with quantitative methods to measure craniofacial structures in developing embryos, we establish the tissue-specific contributions of ephrin-B1 mosaicism to craniofacial dysmorphology. We also examine when ephrin-B1 regulates cellular position during different stages of craniofacial development and which EphB receptors are involved. Our results reveal the specific cellular context and signaling interactions that are likely to underlie CFNS, and provide new understanding of how EPHRIN-B1 may regulate normal craniofacial development.
]]></description>
<dc:creator>Niethamer, T. K.</dc:creator>
<dc:creator>Percival, C. J.</dc:creator>
<dc:creator>Teng, T.</dc:creator>
<dc:creator>Franco, M.</dc:creator>
<dc:creator>Du, Y. X.</dc:creator>
<dc:creator>Bush, J. O.</dc:creator>
<dc:date>2019-07-16</dc:date>
<dc:identifier>doi:10.1101/704619</dc:identifier>
<dc:title><![CDATA[Aberrant cell segregation in craniofacial primordia and the emergence of facial dysmorphology in craniofrontonasal syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/704882v1?rss=1">
<title>
<![CDATA[
Mechanistic Insights into the Protective Roles of Polyphosphate Against Amyloid Cytotoxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/704882v1?rss=1</link>
<description><![CDATA[
The universally abundant polyphosphate (polyP) accelerates fibril formation of disease-related amyloids and protects against amyloid cytotoxicity. To gain insights into the mechanism(s) by which polyP exerts these effects, we focused on -synuclein, a well-studied amyloid protein, which constitutes the major component of Lewy bodies found in Parkinsons Disease. Here we demonstrate that polyP is unable to accelerate the rate-limiting step of -synuclein fibril formation but effectively nucleates fibril assembly once -synuclein oligomers are formed. Binding of polyP to -synuclein either during fibril formation or upon fibril maturation substantially alters fibril morphology, and effectively reduces the ability of -synuclein fibrils to interact with cell membranes. The effect of polyP appears to be -synuclein fibril specific, and successfully prevents the uptake of fibrils into neuronal cells. These results suggest that altering the polyP levels in the extracellular space might be a potential therapeutic strategy to prevent the spreading of the disease.
]]></description>
<dc:creator>Lempart, J.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Lauer, J. A.</dc:creator>
<dc:creator>Ivanova, M. I.</dc:creator>
<dc:creator>Sutter, A.</dc:creator>
<dc:creator>Yoo, N.</dc:creator>
<dc:creator>Huettemann, P.</dc:creator>
<dc:creator>Southworth, D.</dc:creator>
<dc:creator>Jakob, U.</dc:creator>
<dc:date>2019-07-16</dc:date>
<dc:identifier>doi:10.1101/704882</dc:identifier>
<dc:title><![CDATA[Mechanistic Insights into the Protective Roles of Polyphosphate Against Amyloid Cytotoxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/705285v1?rss=1">
<title>
<![CDATA[
Linkage Disequilibrium and Heterozygosity Modulate the Genetic Architecture of Human Complex Phenotypes: Evidence of Natural Selection from GWAS Summary Statistics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/705285v1?rss=1</link>
<description><![CDATA[
We propose an extended Gaussian mixture model for the distribution of causal effects of common single nucleotide polymorphisms (SNPs) for human complex phenotypes that depends on linkage disequilibrium (LD) and heterozygosity (H), while also allowing for independent components for small and large effects. Using a precise methodology showing how genome-wide association studies (GWAS) summary statistics (z-scores) arise through LD with underlying causal SNPs, we applied the model to GWAS of multiple human phenotypes. Our findings indicated that causal effects are distributed with dependence on total LD and H, whereby SNPs with lower total LD and H are more likely to be causal with larger effects; this dependence is consistent with models of the influence of negative pressure from natural selection. Compared with the basic Gaussian mixture model it is built on, the extended model - primarily through quantification of selection pressure - reproduces with greater accuracy the empirical distributions of z-scores, thus providing better estimates of genetic quantities, such as polygenicity and heritability, that arise from the distribution of causal effects.
]]></description>
<dc:creator>Holland, D.</dc:creator>
<dc:creator>Frei, O.</dc:creator>
<dc:creator>Desikan, R.</dc:creator>
<dc:creator>Fan, C. C.</dc:creator>
<dc:creator>Shadrin, A.</dc:creator>
<dc:creator>Smeland, O.</dc:creator>
<dc:creator>Andreassen, O. A.</dc:creator>
<dc:creator>Dale, A.</dc:creator>
<dc:date>2019-07-16</dc:date>
<dc:identifier>doi:10.1101/705285</dc:identifier>
<dc:title><![CDATA[Linkage Disequilibrium and Heterozygosity Modulate the Genetic Architecture of Human Complex Phenotypes: Evidence of Natural Selection from GWAS Summary Statistics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/713545v1?rss=1">
<title>
<![CDATA[
Targeting DNA damage response and Replication Stress in Pancreatic Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/713545v1?rss=1</link>
<description><![CDATA[
Continuing recalcitrance to therapy cements pancreatic cancer (PC) as the most lethal malignancy, which is set to become the second leading cause of cancer death in our society. We interrogated the transcriptome, genome, proteome and functional characteristics of 61 novel PC patient-derived cell lines to define novel therapeutic strategies targeting the DNA damage response (DDR) and replication stress. We show that patient-derived cell lines faithfully recapitulate the epithelial component of pancreatic tumors including previously described molecular subtypes. Biomarkers of DDR deficiency, including a novel signature of homologous recombination deficiency, co-segregates with response to platinum and PARP inhibitor therapy in vitro and in vivo. We generated a novel signature of replication stress with potential clinical utility in predicting response to ATR and WEE1 inhibitor treatment. Replication stress and DDR deficiency are independent of each other, creating opportunities for therapy in DDR proficient PC, and post-platinum therapy.nnnnO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=145 SRC="FIGDIR/small/713545v1_ufig1.gif" ALT="Figure 1">nView larger version (47K):norg.highwire.dtl.DTLVardef@106f2b8org.highwire.dtl.DTLVardef@1294d22org.highwire.dtl.DTLVardef@86f5d3org.highwire.dtl.DTLVardef@751e13_HPS_FORMAT_FIGEXP  M_FIG C_FIG STATEMENT OF SIGNIFICANCEWe define therapeutic strategies that target subgroups of PC using novel signatures of DNA damage response deficiency and replication stress. This potentially offers patients with DNA repair defects therapeutic options outside standard of care platinum chemotherapy and is being tested in clinical trials on the Precision-Panc platform.
]]></description>
<dc:creator>Dreyer, S.</dc:creator>
<dc:creator>Upstill-Goddard, R.</dc:creator>
<dc:creator>Paulus-Hock, V.</dc:creator>
<dc:creator>Paris, C.</dc:creator>
<dc:creator>Maria-Lampraki, E.</dc:creator>
<dc:creator>Dray, E.</dc:creator>
<dc:creator>Serrels, B.</dc:creator>
<dc:creator>Rebus, S.</dc:creator>
<dc:creator>Brunton, H.</dc:creator>
<dc:creator>Nourse, C.</dc:creator>
<dc:creator>Bailey, U.-M.</dc:creator>
<dc:creator>Jones, M.</dc:creator>
<dc:creator>Moran-Jones, K.</dc:creator>
<dc:creator>Wright, D.</dc:creator>
<dc:creator>Duthie, F.</dc:creator>
<dc:creator>Oien, K.</dc:creator>
<dc:creator>Evers, L.</dc:creator>
<dc:creator>McKay, C.</dc:creator>
<dc:creator>McGregor, G.</dc:creator>
<dc:creator>Gulati, A.</dc:creator>
<dc:creator>Brough, R.</dc:creator>
<dc:creator>Bajrami, I.</dc:creator>
<dc:creator>Pettitt, S.</dc:creator>
<dc:creator>Dzubiunski, M.</dc:creator>
<dc:creator>Candido, J.</dc:creator>
<dc:creator>Balkwill, F.</dc:creator>
<dc:creator>Barry, S.</dc:creator>
<dc:creator>Grutzmann, R.</dc:creator>
<dc:creator>Rahib, L.</dc:creator>
<dc:creator>Glasgow Precision Oncology Laboratory,</dc:creator>
<dc:creator>Australian Pancreatic Genome Initiative,</dc:creator>
<dc:creator>Johns, A.</dc:creator>
<dc:creator>Pajic, M.</dc:creator>
<dc:creator>Froeling, F.</dc:creator>
<dc:creator>Beer, P.</dc:creator>
<dc:creator>Musgrove, E.</dc:creator>
<dc:creator>Petersen, G.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Frame, M.</dc:creator>
<dc:creator>Crawford, H.</dc:creator>
<dc:creator>Simeone, D.</dc:creator>
<dc:creator>Lord,</dc:creator>
<dc:date>2019-07-24</dc:date>
<dc:identifier>doi:10.1101/713545</dc:identifier>
<dc:title><![CDATA[Targeting DNA damage response and Replication Stress in Pancreatic Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/715474v1?rss=1">
<title>
<![CDATA[
Recognition of histone H3 methylation states by the PHD1 domain of histone demethylase KDM5A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/715474v1?rss=1</link>
<description><![CDATA[
PHD reader domains are chromatin binding modules often responsible for the recruitment of large protein complexes that contain histone modifying enzymes, chromatin remodelers and DNA repair machinery. A majority of PHD domains recognize N-terminal residues of histone H3 and are sensitive to the methylation state of Lys4 in histone H3 (H3K4). Histone demethylase KDM5A, an epigenetic eraser enzyme that contains three PHD domains, is often overexpressed in various cancers and its demethylation activity is allosterically enhanced when its PHD1 domain is bound to the H3 tail. The allosteric regulatory function of PHD1 expands roles of reader domains, suggesting unique features of this chromatin interacting module. Our previous studies determined the H3 binding site of PHD1, although it remains unclear how the H3 tail interacts with the N-terminal residues of PHD1 and how PHD1 discriminates against H3 tails with varying degrees of H3K4 methylation. Here we have determined the solution structure of apo and H3 bound PHD1. We observe conformational changes occurring in PHD1 in order to accommodate H3, which interestingly binds in a helical conformation. We also observe differential interactions of binding residues with differently methylated H3K4 peptides (me0, me1, me2 or me3), providing a rational for this PHD1 domains preference for lower methylation states of H3K4. We further assessed the contributions of various H3 interacting residues in the PHD1 domain to the binding of H3 peptides. The structural information of the H3 binding site could provide useful information to aid development of allosteric small molecule modulators of KDM5A.
]]></description>
<dc:creator>Longbotham, J. E.</dc:creator>
<dc:creator>Kelly, M. J. S.</dc:creator>
<dc:creator>Galonic Fujmori, D.</dc:creator>
<dc:date>2019-07-26</dc:date>
<dc:identifier>doi:10.1101/715474</dc:identifier>
<dc:title><![CDATA[Recognition of histone H3 methylation states by the PHD1 domain of histone demethylase KDM5A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/716720v1?rss=1">
<title>
<![CDATA[
State and trait characteristics of anterior insula time-varying functional connectivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/716720v1?rss=1</link>
<description><![CDATA[
The human anterior insula (aINS) is a topographically organized brain region, in which ventral portions contribute to socio-emotional function through limbic and autonomic connections, whereas the dorsal aINS contributes to cognitive processes through frontal and parietal connections. Open questions remain, however, regarding how aINS connectivity varies over time. We implemented a novel approach combining seed-to-whole-brain sliding-window functional connectivity MRI and k-means clustering to assess time-varying functional connectivity of aINS subregions. We studied three independent large samples of healthy participants and longitudinal datasets to assess inter- and intra-subject stability, and related aINS time-varying functional connectivity profiles to dispositional empathy. We identified four robust aINS time-varying functional connectivity modes that displayed both "state" and "trait" characteristics: while modes featuring connectivity to sensory regions were modulated by eye closure, modes featuring connectivity to higher cognitive and emotional processing regions were stable over time and related to empathy measures.
]]></description>
<dc:creator>Pasquini, L.</dc:creator>
<dc:creator>Toller, G.</dc:creator>
<dc:creator>Staffaroni, A.</dc:creator>
<dc:creator>Brown, J. A.</dc:creator>
<dc:creator>Deng, J.</dc:creator>
<dc:creator>Lee, A.</dc:creator>
<dc:creator>Kurcyus, K.</dc:creator>
<dc:creator>Shdo, S. M.</dc:creator>
<dc:creator>Allen, I.</dc:creator>
<dc:creator>Sturm, V. E.</dc:creator>
<dc:creator>Cobigo, Y.</dc:creator>
<dc:creator>Borghesani, V.</dc:creator>
<dc:creator>Battistella, G.</dc:creator>
<dc:creator>Gorno-Tempini, M. L.</dc:creator>
<dc:creator>Rankin, K. P.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Rosen, H. H.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:date>2019-07-27</dc:date>
<dc:identifier>doi:10.1101/716720</dc:identifier>
<dc:title><![CDATA[State and trait characteristics of anterior insula time-varying functional connectivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/720565v1?rss=1">
<title>
<![CDATA[
Telomeric Double Strand Breaks Facilitate Formation of 5’ C-Rich Overhangs in G1 Human Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/720565v1?rss=1</link>
<description><![CDATA[
Telomeres are repetitive nucleoprotein complexes that protect chromosomal termini and prevent them from activating an inappropriate DNA damage response (DDR). Here, we characterized the human cellular response to targeted telomeric DSBs in telomerase positive and telomerase-independent alternative lengthening of telomeres (ALT) cells, specifically in G1. Telomeric DSBs in G1 human cells elicited early signatures of a DDR, however, localization of 53BP1, an important regulator of resection at broken ends, was not observed at telomeric break sites. Consistent with this finding and previously reported repression of classical nonhomologous end-joining (c-NHEJ) at telomeres, evidence for c-NHEJ was also lacking. Likewise, no evidence of homologous recombination (HR)-dependent repair of telomeric DSBs in G1 was observed. Rather, and supportive of rapid truncation events, telomeric DSBs in G1 human cells facilitated formation of extensively resected tracks of 5 C-rich telomeric single-stranded (ss)DNA, a previously proposed marker of the recombination dependent ALT pathway. Indeed, induction of telomeric DSBs in human ALT cells also resulted in significant increases in 5 C-rich (ss)telomeric DNA in G1, which rather than RPA, were bound by the complementary telomeric RNA, TERRA. These results suggest that targeting TERRA-mediated protection at damaged telomeres may represent a promising therapeutic strategy, particularly against ALT-positive cancers.
]]></description>
<dc:creator>Nelson, C. B.</dc:creator>
<dc:creator>Alturki, T.</dc:creator>
<dc:creator>Taylor, L.</dc:creator>
<dc:creator>Maranon, D. G.</dc:creator>
<dc:creator>Muraki, K.</dc:creator>
<dc:creator>Murnane, J. P.</dc:creator>
<dc:creator>Bailey, S. M.</dc:creator>
<dc:date>2019-08-01</dc:date>
<dc:identifier>doi:10.1101/720565</dc:identifier>
<dc:title><![CDATA[Telomeric Double Strand Breaks Facilitate Formation of 5’ C-Rich Overhangs in G1 Human Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/730523v1?rss=1">
<title>
<![CDATA[
Structure-function analysis reveals a DNA polymerization-independent role for mitochondrial DNA polymerase IC in African trypanosomes. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/730523v1?rss=1</link>
<description><![CDATA[
The mitochondrial DNA of Trypanosoma brucei and related parasites is a catenated network containing thousands of minicircles and tens of maxicircles called kinetoplast DNA (kDNA). Replication of the single nucleoid requires at least three DNA polymerases (POLIB, POLIC, and POLID) each having discrete localization near the kDNA during S phase. POLIB and POLID have roles in minicircle replication while the specific role of POLIC in kDNA maintenance is less clear. Here, we use an RNAi-complementation system to dissect the functions of the distinct POLIC domains: the conserved family A DNA polymerase domain (POLA) and the uncharacterized N-terminal region (UCR). While RNAi complementation with wild-type POLIC restored kDNA content and cell cycle localization, active site point mutations in the POLA domain impaired minicircle replication similarly to POLIB and POLID depletions. Complementation with the POLA domain alone abolished POLIC foci formation and partially rescued the RNAi phenotype. Furthermore, we provide evidence of a crucial role for the UCR in cell cycle localization and segregation of kDNA daughter networks. This is the first report of a DNA polymerase that impacts DNA segregation.nnSummary statementMitochondrial DNA segregation in African trypanosomes is supported by a dual-functioning DNA polymerase.
]]></description>
<dc:creator>Miller, J. C.</dc:creator>
<dc:creator>Delzell, S. B.</dc:creator>
<dc:creator>Concepcion-Acevedo, J.</dc:creator>
<dc:creator>Boucher, M. J.</dc:creator>
<dc:creator>Klingbeil, M. M.</dc:creator>
<dc:date>2019-08-09</dc:date>
<dc:identifier>doi:10.1101/730523</dc:identifier>
<dc:title><![CDATA[Structure-function analysis reveals a DNA polymerization-independent role for mitochondrial DNA polymerase IC in African trypanosomes.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/736231v1?rss=1">
<title>
<![CDATA[
BET proteins are required for transcriptional activation of the senescent beta cell secretome in Type 1 Diabetes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/736231v1?rss=1</link>
<description><![CDATA[
OBJECTIVEType 1 Diabetes (T1D) results from progressive loss of pancreatic beta cells due to autoimmune destruction. We recently reported that during the natural history of T1D in humans and the female nonobese diabetic (NOD) mouse model, beta cells acquire a senescence-associated secretory phenotype (SASP) that is a major driver of disease onset and progression, but the mechanisms that activate SASP in beta cells were not explored. The objective of this study was to identify transcriptional mechanisms of SASP activation in beta cells.nnMETHODSWe used the female NOD mouse model of spontaneous autoimmune T1D and ex vivo experiments on NOD mouse and human islets to test the hypothesis that Bromodomain Extra-Terminal domain (BET) proteins activate the beta cell SASP transcriptional program.nnRESULTSHere we show that beta cell SASP is transcriptionally controlled by BET proteins, including BRD4. Chromatin analysis of key beta cell SASP genes in NOD islets revealed binding of BRD4 at active regulatory regions. BET protein inhibition in NOD islets diminished not only the transcriptional activation and secretion of SASP factors but also the non-cell autonomous activity. BET protein inhibition also decreased the extent of SASP induction in human islets exposed to DNA damage. The BET protein inhibitor iBET-762 prevented diabetes in NOD mice and also attenuated SASP in beta cells in vivo.nnCONCLUSIONSTaken together, our findings support a crucial role for BET proteins in the activation of the beta cell SASP transcriptional program. These studies suggest avenues for preventing T1D by transcriptional inhibition of SASP.
]]></description>
<dc:creator>Thompson, P. J.</dc:creator>
<dc:creator>Shah, A.</dc:creator>
<dc:creator>Apostolopolou, C.-C.</dc:creator>
<dc:creator>Bhushan, A.</dc:creator>
<dc:date>2019-08-15</dc:date>
<dc:identifier>doi:10.1101/736231</dc:identifier>
<dc:title><![CDATA[BET proteins are required for transcriptional activation of the senescent beta cell secretome in Type 1 Diabetes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/739185v1?rss=1">
<title>
<![CDATA[
Aging induces aberrant state transition kinetics in murine muscle stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/739185v1?rss=1</link>
<description><![CDATA[
Murine muscle stem cells (MuSCs) experience a transition from quiescence to activation that is required for regeneration, but it remains unclear if the transition states and rates of activation are uniform across cells, or how features of this process may change with age. Here, we use timelapse imaging and single cell RNA-seq to measure activation trajectories and rates in young and aged MuSCs. We find that the activation trajectory is conserved in aged cells, and develop effective machine learning classifiers for cell age. Using cell behavior analysis and RNA velocity, we find that activation kinetics are delayed in aged MuSCs, suggesting that changes in stem cell dynamics may contribute to impaired stem cell function with age. Intriguingly, we also find that stem cell activation appears to be a random walk like process, with frequent reversals, rather than a continuous, linear progression. These results support a view of the aged stem cell phenotype as a combination of differences in the location of stable cell states and differences in transition rates between them.nnSummary StatementWe find that aged muscle stem cells display delayed activation dynamics, but retain a youthful activation trajectory, suggesting that changes to cell state dynamics may contribute to aging pathology.
]]></description>
<dc:creator>Kimmel, J. C.</dc:creator>
<dc:creator>Hwang, A. B.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:creator>Brack, A. S.</dc:creator>
<dc:date>2019-08-19</dc:date>
<dc:identifier>doi:10.1101/739185</dc:identifier>
<dc:title><![CDATA[Aging induces aberrant state transition kinetics in murine muscle stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/741983v1?rss=1">
<title>
<![CDATA[
The Interaction of LILRB2 with HLA-B is Associated with Psoriasis Susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/741983v1?rss=1</link>
<description><![CDATA[
Genetic variation within the major histocompatibility complex (MHC) class I is a well-known risk factor for psoriasis. While the mechanisms behind this variation are still being fully elucidated, human leukocyte antigen (HLA) presentation of auto-antigens as well as the interaction of HLA-B with killer cell immunoglobulin-like receptors (KIRs) have been shown to contribute to psoriasis susceptibility. Here we demonstrate that the interaction of HLA class I molecules with leukocyte immunoglobulin-like receptors (LILR), a related group of immunomodulatory receptors primarily found on antigen presenting cells, also contributes to psoriasis susceptibility. We used previously characterized binding capacities of HLA-A, HLA-B, and HLA-C allotypes to two inhibitory LILRs, LILRB1 and LILRB2, to investigate the effect of LILRB1/2 binding in two large genome wide association study cohorts of psoriasis patients and controls (N = 10,069). We found that the strength of binding of LILRB2 to HLA-B was inversely associated with psoriasis risk (p = 2.34E-09, OR [95% CI], 0.41 [0.30-0.55]) independent of individual class I or II allelic effects. We thus propose that weak binding of inhibitory LILRB2 to HLA-B may play a role in patient susceptibility to psoriasis via increased activity of antigen presenting cells.
]]></description>
<dc:creator>Yanovsky, R. L.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Leslie, S.</dc:creator>
<dc:creator>Carrington, M.</dc:creator>
<dc:creator>Liao, W.</dc:creator>
<dc:date>2019-08-21</dc:date>
<dc:identifier>doi:10.1101/741983</dc:identifier>
<dc:title><![CDATA[The Interaction of LILRB2 with HLA-B is Associated with Psoriasis Susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/751990v1?rss=1">
<title>
<![CDATA[
Speed and diffusion of kinesin-2 are competing limiting factors in flagellar length control model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/751990v1?rss=1</link>
<description><![CDATA[
Flagellar length control in Chlamydomonas is a tractable model system for studying the general question of organelle size regulation. We have previously proposed that diffusive return of the kinesin motor that powers intraflagellar transport can play a key role in length regulation. Here we explore how the motor speed and diffusion coefficient for the return of kinesin-2 affect flagellar growth kinetics. We find that the system can exist in two distinct regimes, one dominated by motor speed and one by diffusion coefficient. Depending on length, a flagellum can switch between these regimes. Our results indicate that mutations can affect length in distinct ways. We discuss our theorys implication for flagellar growth influenced by beating and provide possible explanations for the experimental observation that a beating flagellum is usually longer than its immotile mutant. These results demonstrate how our simple model can suggest explanations for mutant phenotypes.nnStatement of SignificanceThe eukaryotic flagellum is an ideal case study in organelle size control because of its simple linear shape and well-understood building mechanism. In our previous work, we proved that flagellar length in the green algae Chlamydomonas can be controlled by the diffusive gradient of the kinesin-2 motors that deliver building blocks to the tip. In this study, we expand on the analytical formulation of the diffusion model to show how physical parameters affect final length and regeneration time, enhancing the models potential to explain length mutants and motivate future research with precise predictions.
]]></description>
<dc:creator>Ma, R.</dc:creator>
<dc:creator>Hendel, N. L.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:creator>Qin, H.</dc:creator>
<dc:date>2019-08-30</dc:date>
<dc:identifier>doi:10.1101/751990</dc:identifier>
<dc:title><![CDATA[Speed and diffusion of kinesin-2 are competing limiting factors in flagellar length control model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/755793v1?rss=1">
<title>
<![CDATA[
Loss of a novel striated muscle-enriched mitochondrial protein Coq10a enhances postnatal cardiac hypertrophic growth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/755793v1?rss=1</link>
<description><![CDATA[
Postnatal mammalian cardiomyocytes undergo a major transition from hyperplasia (increases in cell numbers) to hypertrophy (expansion in cell size). This process is accompanied by rapid mitochondrial biogenesis and metabolic switches to meet the demand of increased cardiac output. Although most mitochondrial components express ubiquitously, recent transcriptomic and proteomic analyses have discovered numerous tissue-specific mitochondrial proteins whose physiological functions are largely unknown. Here we report that a highly evolutionarily conserved mitochondrial protein Coq10a is predominantly expressed in mammalian cardiac and skeletal muscles, and is highly up-regulated around birth in a thyroid hormone-dependent manner. Deletion of Coq10a by CRISPR/Cas9 leads to enhanced cardiac growth after birth. Surprisingly, adult Coq10a mutant mice maintain the hypertrophic heart phenotype with increased levels of coenzyme Q (CoQ) per cardiomyocyte, preserved cardiac contractile function and mitochondrial respiration, which contrasts with reported mice and humans with mutations in other Coq family genes. Further RNA-seq analysis and mitochondrial characterization suggest an increase of mitochondrial biogenesis in the Coq10a mutant heart as a possible consequence of Peroxisome proliferator-activated receptor Gamma Coactivator 1-alpha (PGC1) activation, consistent with a recent intriguing report that CoQ may function as a natural ligand and partial agonist of Peroxisome Proliferator-Activated Receptor (PPAR) /{gamma}. Taken together, our study reveals a previously unknown function of a novel striated muscle-enriched mitochondrial protein Coq10a in regulating postnatal heart growth.
]]></description>
<dc:creator>Hirose, K.</dc:creator>
<dc:creator>Chang, S.</dc:creator>
<dc:creator>Yu, H.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Barca, E.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Khyeam, S.</dc:creator>
<dc:creator>Tajima, K.</dc:creator>
<dc:creator>Yoneshiro, T.</dc:creator>
<dc:creator>Kajimura, S.</dc:creator>
<dc:creator>Quinzii, C. M.</dc:creator>
<dc:creator>Hu, G.</dc:creator>
<dc:creator>Huang, G. N.</dc:creator>
<dc:date>2019-09-03</dc:date>
<dc:identifier>doi:10.1101/755793</dc:identifier>
<dc:title><![CDATA[Loss of a novel striated muscle-enriched mitochondrial protein Coq10a enhances postnatal cardiac hypertrophic growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/755827v1?rss=1">
<title>
<![CDATA[
In vitro and in vivo roles of glucocorticoid and vitamin D receptors in the control of cardiomyocyte proliferative potential 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/755827v1?rss=1</link>
<description><![CDATA[
Cardiomyocyte (CM) proliferative potential varies considerably across species. While lower vertebrates and neonatal mammals retain robust capacities for CM proliferation, adult mammalian CMs lose proliferative potential due to cell-cycle withdrawal and polyploidization, failing to mount a proliferative response to regenerate lost CMs after cardiac injury. The decline of murine CM proliferative potential occurs in the neonatal period when the endocrine system undergoes drastic changes for adaptation to extrauterine life. We recently demonstrated that thyroid hormone (TH) signaling functions as a primary factor driving CM proliferative potential loss in vertebrates. Whether other hormonal pathways govern this process remains largely unexplored. Here we showed that agonists of glucocorticoid receptor (GR) and vitamin D receptor (VDR) suppressed neonatal CM proliferation in vitro. We next examined CM nucleation and proliferation in mutant mice lacking GR or VDR specifically in CMs, but we observed no difference between mutant and control littermates. Additionally, we generated compound mutant mice that lack GR or VDR and express dominant-negative TH receptor alpha in their CMs, and similarly observed no increase in CM proliferative potential compared to dominant-negative TH receptor alpha mice alone. Thus, although GR and VDR activation in cultured CMs is sufficient to inhibit CM proliferation, they seem to be dispensable for CM cell-cycle exit and binucleation in vivo. In addition, given the recent report that VDR activation in zebrafish promotes CM proliferation and tissue regeneration, our results suggest distinct roles of VDR in zebrafish and rodent CM cell-cycle regulation.
]]></description>
<dc:creator>Cutie, S.</dc:creator>
<dc:creator>Lunn, D.</dc:creator>
<dc:creator>Huang, G.</dc:creator>
<dc:date>2019-09-03</dc:date>
<dc:identifier>doi:10.1101/755827</dc:identifier>
<dc:title><![CDATA[In vitro and in vivo roles of glucocorticoid and vitamin D receptors in the control of cardiomyocyte proliferative potential]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/755991v1?rss=1">
<title>
<![CDATA[
Defined factors to reactivate cell cycle activity in adult mouse cardiomyocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/755991v1?rss=1</link>
<description><![CDATA[
Adult mammalian cardiomyocytes exit the cell cycle during the neonatal period, commensurate with the loss of regenerative capacity in adult mammalian hearts. We established conditions for long-term culture of adult mouse cardiomyocytes that are genetically labeled with fluorescence. This technique permits reliable analyses of proliferation of pre-existing cardiomyocytes without complications from cardiomyocyte marker expression loss due to dedifferentiation or significant contribution from cardiac progenitor cell expansion and differentiation in culture. Using this system, we took a candidate gene approach to screen for fetal-specific proliferative gene programs that can induce proliferation of adult mouse cardiomyocytes. Using pooled gene delivery and subtractive gene elimination, we identified a novel functional interaction between E2f Transcription Factor 2 (E2f2) and Brain Expressed X-Linked (Bex)/Transcription elongation factor A-like (Tceal) superfamily members Bex1 and Tceal8. Specifically, Bex1 and Tceal8 both preserved cell viability during E2f2-induced cell cycle re-entry. Although Tceal8 inhibited E2f2-induced S-phase re-entry, Bex1 facilitated DNA synthesis while inhibiting cell death. In sum, our study provides a valuable method for adult cardiomyocyte proliferation research and suggests that Bex family proteins may function in modulating cell proliferation and death decisions during cardiomyocyte development and maturation.
]]></description>
<dc:creator>Judd, J.</dc:creator>
<dc:creator>Lovas, J.</dc:creator>
<dc:creator>Huang, G. N.</dc:creator>
<dc:date>2019-09-03</dc:date>
<dc:identifier>doi:10.1101/755991</dc:identifier>
<dc:title><![CDATA[Defined factors to reactivate cell cycle activity in adult mouse cardiomyocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/761718v1?rss=1">
<title>
<![CDATA[
Complement component 4 genes contribute sex-specific vulnerability in diverse illnesses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/761718v1?rss=1</link>
<description><![CDATA[
Many common illnesses differentially affect men and women for unknown reasons. The autoimmune diseases lupus and Sjogrens syndrome affect nine times more women than men1,2, whereas schizophrenia affects men more frequently and severely3-5. All three illnesses have their strongest common-genetic associations in the Major Histocompatibility Complex (MHC) locus, an association that in lupus and Sjogrens syndrome has long been thought to arise from HLA alleles6-13. Here we show that the complement component 4 (C4) genes in the MHC locus, recently found to increase risk for schizophrenia14, generate 7-fold variation in risk for lupus (95% CI: 5.88-8.61; p < 10-117 in total) and 16-fold variation in risk for Sjogrens syndrome (95% CI: 8.59-30.89; p < 10-23 in total), with C4A protecting more strongly than C4B in both illnesses. The same alleles that increase risk for schizophrenia, greatly reduced risk for lupus and Sjogrens syndrome. In all three illnesses, C4 alleles acted more strongly in men than in women: common combinations of C4A and C4B generated 14-fold variation in risk for lupus and 31-fold variation in risk for Sjogrens syndrome in men (vs. 6-fold and 15-fold among women respectively) and affected schizophrenia risk about twice as strongly in men as in women. At a protein level, both C4 and its effector (C3) were present at greater levels in men than women in cerebrospinal fluid (p < 10-5 for both C4 and C3) and plasma among adults ages 20-5015-17, corresponding to the ages of differential disease vulnerability. Sex differences in complement protein levels may help explain the larger effects of C4 alleles in men, womens greater risk of SLE and Sjogrens, and mens greater vulnerability in schizophrenia. These results nominate the complement system as a source of sexual dimorphism in vulnerability to diverse illnesses.
]]></description>
<dc:creator>Kamitaki, N.</dc:creator>
<dc:creator>Sekar, A.</dc:creator>
<dc:creator>Handsaker, R. E.</dc:creator>
<dc:creator>de Rivera, H.</dc:creator>
<dc:creator>Tooley, K.</dc:creator>
<dc:creator>Morris, D. L.</dc:creator>
<dc:creator>Taylor, K. E.</dc:creator>
<dc:creator>Whelan, C. W.</dc:creator>
<dc:creator>Tombleson, P.</dc:creator>
<dc:creator>Olde Loohuis, L. M.</dc:creator>
<dc:creator>Schizophrenia Working Group of the Psychiatric Genomics Consortium,</dc:creator>
<dc:creator>Boehnke, M.</dc:creator>
<dc:creator>Kimberly, R. P.</dc:creator>
<dc:creator>Kaufman, K. M.</dc:creator>
<dc:creator>Harley, J. B.</dc:creator>
<dc:creator>Langefeld, C. D.</dc:creator>
<dc:creator>Seidman, C. E.</dc:creator>
<dc:creator>Pato, M. T.</dc:creator>
<dc:creator>Pato, C. N.</dc:creator>
<dc:creator>Ophoff, R. A.</dc:creator>
<dc:creator>Graham, R. R.</dc:creator>
<dc:creator>Criswell, L. A.</dc:creator>
<dc:creator>Vyse, T. J.</dc:creator>
<dc:creator>McCarroll, S. A.</dc:creator>
<dc:date>2019-09-09</dc:date>
<dc:identifier>doi:10.1101/761718</dc:identifier>
<dc:title><![CDATA[Complement component 4 genes contribute sex-specific vulnerability in diverse illnesses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/762336v1?rss=1">
<title>
<![CDATA[
Dissecting the chitinolytic activity of mammalian chitinases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/762336v1?rss=1</link>
<description><![CDATA[
Chitin is an abundant polysaccharide used by a large range of organisms for structural rigidity and water repulsion. As such, the insoluble crystalline structure of chitin poses significant challenges for enzymatic degradation. Vertebrates do not produce chitin, but do express chitin degrading enzymes. Acidic mammalian chitinase, the primary enzyme involved in the degradation of environmental chitin in mammalian lungs, is a processive glycosyl hydrolase that may be able to make multiple hydrolysis events for each binding event. Mutations to acidic mammalian chitinase have been associated with asthma, and genetic deletion of the enzyme in mice results in significantly increased morbidity and mortality with age. We initially set out to reverse this phenotype by engineering hyperactive acidic mammalian chitinase variants. Using a directed evolution screening approach using commercial fluorogenic substrates, we identified mutations with consistent increases in activity. To determine whether the activity increases observed with oligomeric substrates were consistent with more biologically relevant chitin substrates, we developed new assays to quantify chitinase activity with colloidal crystalline chitin, and identified a high throughput fluorogenic assay that gives sufficient signal to noise advantages to quantify changes to activity due to the addition or removal of a chitin binding domain to the enzyme. We show that the activity increasing mutations derived from our directed evolution screen were lost when crystalline substrates were used. In contrast, naturally occurring gain-of-function mutations gave similar results with oligomeric and crystalline substrates. We also show that the activity differences between acidic mammalian chitinase and chitotriosidase are reduced in the context of crystalline substrate, suggesting that previously reported activity differences with oligomeric substrates may have been largely driven by differential substrate specificity for the oligomers. These results highlight the need for assays against more physiological substrates when engineering complex metabolic enzymes, and provide a new approach that may be broadly applicable to engineering glycosyl hydrolases.
]]></description>
<dc:creator>Barad, B. A.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Diaz, R. E.</dc:creator>
<dc:creator>Basilio, R.</dc:creator>
<dc:creator>Van Dyken, S. J.</dc:creator>
<dc:creator>Locksley, R. M.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2019-09-08</dc:date>
<dc:identifier>doi:10.1101/762336</dc:identifier>
<dc:title><![CDATA[Dissecting the chitinolytic activity of mammalian chitinases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/764118v1?rss=1">
<title>
<![CDATA[
Transcriptomic profiling of plaque psoriasis and cutaneous T cell subsets during treatment with secukinumab 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/764118v1?rss=1</link>
<description><![CDATA[
The IL17A inhibitor secukinumab is efficacious for the treatment of psoriasis. In order to define its mechanism of action, it is important to understand its impact on psoriatic whole skin tissue as well as specific skin-resident immune cell populations such as T lymphocytes. In this study, we treated 15 moderate-to-severe plaque psoriasis patients with secukinumab and characterized the longitudinal transcriptomic changes of whole lesional skin tissue and cutaneous CD4+ T effector cells (Teffs), CD4+ T regulatory cells (Tregs), and CD8+ T effector cells during 12 weeks of treatment. Secukinumab was clinically effective, with 100%, 47%, and 27% of patients in the study achieving PASI75, PASI90, and PASI100 by week 12, respectively. At baseline prior to treatment, we observed that IL17A overexpression predominates in psoriatic CD8+ T cells rather than Teffs, supporting the importance of IL-17-secreting CD8+ T cells (Tc17) compared to IL-17-secreting CD4+ T cells (Th17) cells in the pathogenesis of psoriasis. Although secukinumab targets only IL17A, we observed rapid reduction of IL17A, IL17F, IL23A, IL23R, and IFNG expression in lesional skin as soon as 2 weeks after initiation of treatment and normalization of expression by week 12. Secukinumab treatment resulted in resolution of 89-97% of psoriasis-associated expression differences in both bulk tissue and T cell subsets by week 12 of treatment. Overall, secukinumab appears to rapidly reverse many of the molecular hallmarks of psoriasis.
]]></description>
<dc:creator>Liu, J. H.</dc:creator>
<dc:creator>Chang, H.-W.</dc:creator>
<dc:creator>Beck, K.</dc:creator>
<dc:creator>Sekhon, S.</dc:creator>
<dc:creator>Schmidt, T. H.</dc:creator>
<dc:creator>Yan, D.</dc:creator>
<dc:creator>Huang, Z.-M.</dc:creator>
<dc:creator>Yang, E.</dc:creator>
<dc:creator>Sanchez, I.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Bhattarai, S.</dc:creator>
<dc:creator>Thibodeaux, Q.</dc:creator>
<dc:creator>Ahn, R. S.</dc:creator>
<dc:creator>Bhutani, T.</dc:creator>
<dc:creator>Rosenblum, M. D.</dc:creator>
<dc:creator>Liao, W.</dc:creator>
<dc:date>2019-09-10</dc:date>
<dc:identifier>doi:10.1101/764118</dc:identifier>
<dc:title><![CDATA[Transcriptomic profiling of plaque psoriasis and cutaneous T cell subsets during treatment with secukinumab]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/766972v1?rss=1">
<title>
<![CDATA[
Optimal design of single-cell RNA sequencing experiments for cell-type-specific eQTL analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/766972v1?rss=1</link>
<description><![CDATA[
Single-cell RNA-sequencing (scRNA-Seq) is a compelling approach to simultaneously measure cellular composition and state which is impossible with bulk profiling approaches. However, it has not yet become a widely used tool in population-scale analyses, due to its prohibitively high cost. Here we show that given the same budget, the statistical power of cell-type-specific expression quantitative trait loci (eQTL) mapping can be increased through low-coverage per-cell sequencing of more samples rather than high-coverage sequencing of fewer samples. We also show that multiple experimental designs with different numbers of samples, cells per sample and reads per cell could have similar statistical power, and choosing an appropriate design can yield large cost savings especially when multiplexed workflows are considered. Finally, we provide a practical approach on selecting cost-effective designs for maximizing cell-type-specific eQTL power.
]]></description>
<dc:creator>Mandric, I.</dc:creator>
<dc:creator>Schwarz, T.</dc:creator>
<dc:creator>Majumdar, A.</dc:creator>
<dc:creator>Perez, R.</dc:creator>
<dc:creator>Subramaniam, M.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Pasaniuc, B.</dc:creator>
<dc:creator>Halperin, E.</dc:creator>
<dc:date>2019-09-12</dc:date>
<dc:identifier>doi:10.1101/766972</dc:identifier>
<dc:title><![CDATA[Optimal design of single-cell RNA sequencing experiments for cell-type-specific eQTL analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/767061v1?rss=1">
<title>
<![CDATA[
Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/767061v1?rss=1</link>
<description><![CDATA[
Innovative new crystallographic methods are facilitating structural studies from ever smaller crystals of biological macromolecules. In particular, serial X-ray crystallography and microcrystal electron diffraction (MicroED) have emerged as useful methods for obtaining structural information from crystals on the nanometer to micron scale. Despite the utility of these methods, their implementation can often be difficult, as they present many challenges not encountered in traditional macromolecular crystallography experiments. Here, we describe XFEL serial crystallography experiments and MicroED experiments using batch-grown microcrystals of the enzyme cyclophilin A (CypA). Our results provide a roadmap for researchers hoping to design macromolecular microcrystallography experiments, and they highlight the strengths and weaknesses of the two methods. Specifically, we focus on how the different physical conditions imposed by the sample preparation and delivery methods required for each type of experiment effect the crystal structure of the enzyme.
]]></description>
<dc:creator>Wolff, A. M.</dc:creator>
<dc:creator>Young, I. D.</dc:creator>
<dc:creator>Sierra, R. G.</dc:creator>
<dc:creator>Brewster, A. S.</dc:creator>
<dc:creator>Martynowycz, M. W.</dc:creator>
<dc:creator>Nango, E.</dc:creator>
<dc:creator>Sugahara, M.</dc:creator>
<dc:creator>Nakane, T.</dc:creator>
<dc:creator>Ito, K.</dc:creator>
<dc:creator>Aquila, A.</dc:creator>
<dc:creator>Bhowmick, A.</dc:creator>
<dc:creator>Biel, J. T.</dc:creator>
<dc:creator>Carbajo, S.</dc:creator>
<dc:creator>Cohen, A. E.</dc:creator>
<dc:creator>Cortez, S.</dc:creator>
<dc:creator>Gonzalez, A.</dc:creator>
<dc:creator>Hino, T.</dc:creator>
<dc:creator>Im, D.</dc:creator>
<dc:creator>Koralek, J. D.</dc:creator>
<dc:creator>Kubo, M.</dc:creator>
<dc:creator>Lazarou, T. S.</dc:creator>
<dc:creator>Nomura, T.</dc:creator>
<dc:creator>Owada, S.</dc:creator>
<dc:creator>Samelson, A.</dc:creator>
<dc:creator>Tanaka, R.</dc:creator>
<dc:creator>Tanaka, T.</dc:creator>
<dc:creator>Thompson, E. M.</dc:creator>
<dc:creator>van den Bedem, H.</dc:creator>
<dc:creator>Woldeyes, R. A.</dc:creator>
<dc:creator>Yumoto, F.</dc:creator>
<dc:creator>Zhao, W.</dc:creator>
<dc:creator>Tono, K.</dc:creator>
<dc:creator>Boutet, S.</dc:creator>
<dc:creator>Iwata, S.</dc:creator>
<dc:creator>Gonen, T.</dc:creator>
<dc:creator>Sauter, N. K.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Thompson, M. C.</dc:creator>
<dc:date>2019-09-12</dc:date>
<dc:identifier>doi:10.1101/767061</dc:identifier>
<dc:title><![CDATA[Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/775361v1?rss=1">
<title>
<![CDATA[
Definition of alleles and altered regulatory motifs across Cas9-edited cell populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/775361v1?rss=1</link>
<description><![CDATA[
BackgroundGenetic alteration of candidate response elements at their native chromosomal loci is the only valid determinant of their potential transcriptional regulatory activities. Targeted DNA cleavage by Cas9 coupled with cellular repair processes can produce arrays of alleles that can be defined by massively parallel sequencing by synthesis (SBS), presenting an opportunity to generate and survey edited cell populations that include informative alterations. Such editing efforts commonly rely on subclonal enrichment to isolate cells with preferred genotypic properties at target loci; short nucleotide adducts (indices/barcodes) allow PCR-amplified molecules from diverse sample sources to be pooled, sequenced, and demultiplexed to resolve source-specific content. Not widely available, however, are capabilities for barcoding thousands of clones, or for automated analysis of individual candidate regulatory loci PCR-amplified and sequenced from a genetically heterogeneous population--specifically, imputation of discrete genotype(s) by allele definition and abundance, and identification of altered regulatory factor binding motifs.nnResultsWe describe a panel of 192 8-nucleotide barcode primers compatible with Illumina(R) sequencing platforms, and the application of these barcodes to genotypic analysis of Cas9-edited clones. Permutations of the ninety-six i7 (read 1) and ninety-six i5 (read 2) barcodes allow unique labeling of up to 9,216 distinct samples. We created three independent Python scripts: SampleSheet.py automates construction of Illumina(R) Sample Sheets encoding up to 9,216 barcode:sample relationships; ImputedGenotypes.py defines alleles and imputes genotypes from demultiplexed fastq files; CollatedMotifs.py flags transcription factor recognition motif matches altered in alleles relative to a reference sequence.nnConclusionsCode-enabled definition of alleles and regulatory motifs in sequenced, demultiplexed amplicons facilitates evaluation of genetic diversity in up to 9,216 distinct samples. Here, we demonstrate the utility of three scripts in analysis of cell populations targeted by Cas9 for disruption of glucocorticoid receptor (GR) binding sites near FKBP5, a GR-regulated gene in the human adenocarcinoma cell line A549. SampleSheet.py, ImputedGenotypes.py, and CollatedMotifs.py operate independently and are broadly applicable beyond the case described here.
]]></description>
<dc:creator>Ehmsen, K. T.</dc:creator>
<dc:creator>Knuesel, M. T.</dc:creator>
<dc:creator>Martinez, D.</dc:creator>
<dc:creator>Asahina, M.</dc:creator>
<dc:creator>Aridomi, H.</dc:creator>
<dc:creator>Yamamoto, K. R.</dc:creator>
<dc:date>2019-09-19</dc:date>
<dc:identifier>doi:10.1101/775361</dc:identifier>
<dc:title><![CDATA[Definition of alleles and altered regulatory motifs across Cas9-edited cell populations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/776484v1?rss=1">
<title>
<![CDATA[
Differential modulation of ventral tegmental area circuits by the nociceptin/orphanin FQ system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/776484v1?rss=1</link>
<description><![CDATA[
The neuropeptide nociceptin/orphanin FQ (N/OFQ) can be released by stressors and is associated with disorders of emotion regulation and reward processing. N/OFQ and its receptor, NOP, are enriched in dopaminergic pathways, and intra-ventricular agonist delivery decreases dopamine levels in the dorsal striatum, nucleus accumbens (NAc), and ventral tegmental area (VTA). We used whole cell electrophysiology in acute rat midbrain slices to investigate synaptic actions of N/OFQ. N/OFQ was primarily inhibitory, causing outward currents in both immunocytochemically identified dopaminergic (tyrosine hydroxylase positive (TH(+)) and non-dopaminergic (TH(-)) VTA neurons (effect at 1 M: 20 {+/-} 4 pA). Surprisingly, this effect was mediated by augmentation of postsynaptic GABAAR currents, unlike the substantia nigra pars compacta (SNc), where the N/OFQ induced outward currents were K+ channel dependent. A smaller population, 19% of all VTA neurons, responded to low concentrations N/OFQ with inward currents (10 nM: -11 {+/-} 2 pA). Following 100 nM N/OFQ, the response to a second N/OFQ application was markedly diminished in VTA neurons (14 {+/-} 10% of first response), but not in SNc neurons (90 {+/-} 20% of first response). N/OFQ generated outward currents in medial prefrontal cortex (mPFC)-projecting VTA neurons, but inward currents in a subset of posterior anterior cingulate cortex-projecting VTA neurons. While N/OFQ inhibited NAc-projecting VTA cell bodies, it had little effect on electrically or optogenetically evoked terminal dopamine release in the NAc measured ex vivo with fast scan cyclic voltammetry. These results extend our understanding of the N/OFQ system in brainstem circuits implicated in many neurobehavioral disorders.

Significance statementThe neuropeptide nociceptin/orphanin FQ (N/OFQ) and its receptor (NOP) are engaged under conditions of stress and are associated with reward processing disorders. Both peptide and receptor are highly enriched in ventral tegmental area (VTA) pathways underlying motivation and reward. Using whole cell electrophysiology in rat midbrain slices we found: 1) NOPs are functional on both dopaminergic and non-dopaminergic VTA neurons; 2) N/OFQ differentially regulates VTA neurons based on neuroanatomical projection target; and 3) repeated application of N/OFQ produces evidence of receptor desensitization in the VTA but not the SNc. These results reveal candidate mechanisms by which the NOP system regulates motivation and emotion.
]]></description>
<dc:creator>Driscoll, J. R.</dc:creator>
<dc:creator>Wallace, T. L.</dc:creator>
<dc:creator>Martin, W. J.</dc:creator>
<dc:creator>Margolis, E. B.</dc:creator>
<dc:date>2019-09-19</dc:date>
<dc:identifier>doi:10.1101/776484</dc:identifier>
<dc:title><![CDATA[Differential modulation of ventral tegmental area circuits by the nociceptin/orphanin FQ system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/782748v1?rss=1">
<title>
<![CDATA[
Inherited Causes of Clonal Hematopoiesis of Indeterminate Potential in TOPMed Whole Genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/782748v1?rss=1</link>
<description><![CDATA[
Age is the dominant risk factor for most chronic human diseases; yet the mechanisms by which aging confers this risk are largely unknown.1 Recently, the age-related acquisition of somatic mutations in regenerating hematopoietic stem cell populations was associated with both hematologic cancer incidence2-4 and coronary heart disease prevalence.5 Somatic mutations with leukemogenic potential may confer selective cellular advantages leading to clonal expansion, a phenomenon termed  Clonal Hematopoiesis of Indeterminate Potential (CHIP).6 Simultaneous germline and somatic whole genome sequence analysis now provides the opportunity to identify root causes of CHIP. Here, we analyze high-coverage whole genome sequences from 97,691 participants of diverse ancestries in the NHLBI TOPMed program and identify 4,229 individuals with CHIP. We identify associations with blood cell, lipid, and inflammatory traits specific to different CHIP genes. Association of a genome-wide set of germline genetic variants identified three genetic loci associated with CHIP status, including one locus at TET2 that was African ancestry specific. In silico-informed in vitro evaluation of the TET2 germline locus identified a causal variant that disrupts a TET2 distal enhancer. Aggregates of rare germline loss-of-function variants in CHEK2, a DNA damage repair gene, predisposed to CHIP acquisition. Overall, we observe that germline genetic variation altering hematopoietic stem cell function and the fidelity of DNA-damage repair increase the likelihood of somatic mutations leading to CHIP.
]]></description>
<dc:creator>Bick, A. G.</dc:creator>
<dc:creator>Weinstock, J.</dc:creator>
<dc:creator>Nandakumar, S. K.</dc:creator>
<dc:creator>Fulco, C. P.</dc:creator>
<dc:creator>Leventhal, M. J.</dc:creator>
<dc:creator>Bao, E. L.</dc:creator>
<dc:creator>Nasser, J.</dc:creator>
<dc:creator>Zekavat, S. M.</dc:creator>
<dc:creator>Szeto, M. D.</dc:creator>
<dc:creator>Laurie, C.</dc:creator>
<dc:creator>Taub, M.</dc:creator>
<dc:creator>Mitchell, B.</dc:creator>
<dc:creator>Barnes, K.</dc:creator>
<dc:creator>Moscati, A.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Redline, S.</dc:creator>
<dc:creator>Psaty, B. M.</dc:creator>
<dc:creator>Silverman, E.</dc:creator>
<dc:creator>Weiss, S.</dc:creator>
<dc:creator>Palmer, N.</dc:creator>
<dc:creator>Ramachandran, V.</dc:creator>
<dc:creator>Burchard, E.</dc:creator>
<dc:creator>Kardia, S.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Kaplan, R.</dc:creator>
<dc:creator>Smith, N. L.</dc:creator>
<dc:creator>Arnett, D.</dc:creator>
<dc:creator>Schwartz, D.</dc:creator>
<dc:creator>Correa, A.</dc:creator>
<dc:creator>de Andrade, M.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Konkle, B. A.</dc:creator>
<dc:creator>Custer, B.</dc:creator>
<dc:creator>Peralta, J.</dc:creator>
<dc:creator>Gui, H.</dc:creator>
<dc:creator>Meyers, D.</dc:creator>
<dc:creator>McGarvey, S. T.</dc:creator>
<dc:creator>Chen, I.</dc:creator>
<dc:creator>Shoemaker, M. B.</dc:creator>
<dc:creator>Peyser, P. A.</dc:creator>
<dc:creator>Broome, J.</dc:creator>
<dc:creator>Gogarten, S.</dc:creator>
<dc:creator>Wang, F. F.</dc:creator>
<dc:creator>Wong, Q.</dc:creator>
<dc:creator>Montasser, M.</dc:creator>
<dc:creator>Da</dc:creator>
<dc:date>2019-09-27</dc:date>
<dc:identifier>doi:10.1101/782748</dc:identifier>
<dc:title><![CDATA[Inherited Causes of Clonal Hematopoiesis of Indeterminate Potential in TOPMed Whole Genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/783290v1?rss=1">
<title>
<![CDATA[
Hedgehog pathway activation through conformational blockade of the Patched sterol conduit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/783290v1?rss=1</link>
<description><![CDATA[
Activation of the Hedgehog pathway may have therapeutic value for improved bone healing, taste receptor cell regeneration, and alleviation of colitis or other conditions. Systemic pathway activation, however, may be detrimental and therapeutic application has been difficult for lack of agents amenable to tissue targeting. We have developed a novel agonist, a conformation-specific nanobody against the Hedgehog receptor Patched1. This nanobody potently activates the Hedgehog pathway in vitro and in vivo by stabilizing an alternative conformation of a Patched1 "switch helix", as revealed by cryo-EM structure determination. Although this conformation likely constitutes part of the transport cycle, nanobody-trapping disrupts the cycle and prevents substrate movement through the Patched1 sterol conduit. Our conformation-selective nanobody approach provides a new route to the development of transporter-related pharmacologic agents and may be generally applicable to the study of other transporters.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Lu, W.-J.</dc:creator>
<dc:creator>Bulkley, D.</dc:creator>
<dc:creator>Liang, J.</dc:creator>
<dc:creator>Ralko, A.</dc:creator>
<dc:creator>Roberts, K. J.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Cho, W.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:creator>Beachy, P.</dc:creator>
<dc:date>2019-09-26</dc:date>
<dc:identifier>doi:10.1101/783290</dc:identifier>
<dc:title><![CDATA[Hedgehog pathway activation through conformational blockade of the Patched sterol conduit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/786285v1?rss=1">
<title>
<![CDATA[
Enhancing droplet-based single-nucleus RNA-seq resolution using the semi-supervised machine learning classifier DIEM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/786285v1?rss=1</link>
<description><![CDATA[
Single-nucleus RNA sequencing (snRNA-seq) measures gene expression in individual nuclei instead of cells, allowing for unbiased cell type characterization in solid tissues. Contrary to single-cell RNA seq (scRNA-seq), we observe that snRNA-seq is commonly subject to contamination by high amounts of extranuclear background RNA, which can lead to identification of spurious cell types in downstream clustering analyses if overlooked. We present a novel approach to remove debris-contaminated droplets in snRNA-seq experiments, called Debris Identification using Expectation Maximization (DIEM). Our likelihood-based approach models the gene expression distribution of debris and cell types, which are estimated using EM. We evaluated DIEM using three snRNA-seq data sets: 1) human differentiating preadipocytes in vitro, 2) fresh mouse brain tissue, and 3) human frozen adipose tissue (AT) from six individuals. All three data sets showed various degrees of extranuclear RNA contamination. We observed that existing methods fail to account for contaminated droplets and led to spurious cell types. When compared to filtering using these state of the art methods, DIEM better removed droplets containing high levels of extranuclear RNA and led to higher quality clusters. Although DIEM was designed for snRNA-seq data, we also successfully applied DIEM to single-cell data. To conclude, our novel method DIEM removes debris-contaminated droplets from single-cell-based data fast and effectively, leading to cleaner downstream analysis. Our code is freely available for use at https://github.com/marcalva/diem.
]]></description>
<dc:creator>Alvarez, M.</dc:creator>
<dc:creator>Rahmani, E.</dc:creator>
<dc:creator>Garske, K. M.</dc:creator>
<dc:creator>Miao, Z.</dc:creator>
<dc:creator>Benhammou, J. N.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Pisegna, J. R.</dc:creator>
<dc:creator>Pietilainen, K. H.</dc:creator>
<dc:creator>Halperin, E.</dc:creator>
<dc:creator>Pajukanta, P.</dc:creator>
<dc:date>2019-09-30</dc:date>
<dc:identifier>doi:10.1101/786285</dc:identifier>
<dc:title><![CDATA[Enhancing droplet-based single-nucleus RNA-seq resolution using the semi-supervised machine learning classifier DIEM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/793380v1?rss=1">
<title>
<![CDATA[
Human hepatic tryptophan 2,3-dioxygenase ubiquitin-dependent protein degradation: The critical role of its exosite as the molecular lynchpin of its substrate-mediated protein stabilization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/793380v1?rss=1</link>
<description><![CDATA[
Hepatic tryptophan 2,3-dioxygenase (TDO) is a cytoplasmic homotetrameric hemoprotein and the rate-limiting enzyme in the irreversible degradation of the essential amino acid L-tryptophan (L-Trp) to N-formylkynurenine, thus controlling the flux of L-Trp into its serotonergic and kynureninic/NAD pathways. TDO has long been recognized to be substrate-inducible via protein stabilization, but the molecular mechanism of this stabilization has remained elusive. Recent elucidation of human TDO (hTDO) crystal structure has identified a high-affinity (Kd {approx} 0.5 M) Trp-binding exosite in each of its 4 monomeric subunits. Mutation of the Glu105, Trp208 and Arg211 comprising this exosite not only abolished the high-affinity L-Trp binding, but also accelerated the ubiquitin-dependent proteasomal degradation of hTDO. We have further characterized this hTDO degradation by documenting that its ubiquitination by gp78/AMFR and CHIP E2/E3 ligase complexes occurs on external Lys-residues within or vicinal to acidic Asp/Glu and phosphorylated pSer/pThr (DEpSpT)-clusters. Furthermore, we have identified the unstructured hTDO N- and C-termini as imparting relatively high proteolytic instability, as their deletion ({Delta}NC) markedly prolonged hTDO t1/2. Additionally, although previous studies reported that upon hepatic heme-depletion, the heme-free apoTDO turns over with a t1/2 {approx} 2.2 h relative to the t1/2 of 7.7 h of holoTDO, mutating the axial heme-ligating His328 to Ala has the opposite effect of prolonging hTDO t1/2. Most importantly, introducing the exosite mutation into the {Delta}NC-deleted or H328A-mutant completely abolished their prolonged half-lives irrespective of L-Trp presence or absence, thereby revealing that the exosite is the molecular lynchpin that defines L-Trp-mediated TDO induction via protein stabilization.
]]></description>
<dc:creator>Kim, S.-M.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Karkashon, S.</dc:creator>
<dc:creator>Lewis-Ballester, A.</dc:creator>
<dc:creator>Yeh, S.-R.</dc:creator>
<dc:creator>Correia, M. A.</dc:creator>
<dc:date>2019-10-04</dc:date>
<dc:identifier>doi:10.1101/793380</dc:identifier>
<dc:title><![CDATA[Human hepatic tryptophan 2,3-dioxygenase ubiquitin-dependent protein degradation: The critical role of its exosite as the molecular lynchpin of its substrate-mediated protein stabilization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/794792v1?rss=1">
<title>
<![CDATA[
Estrogen Drives Melanocortin Neurons To Reduce Sedentary Behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/794792v1?rss=1</link>
<description><![CDATA[
Estrogen depletion in rodents and humans leads to inactivity, unhealthy fat accumulation, and diabetes1,2, underscoring the conserved metabolic benefits of estrogen that inevitably decline with aging. In rodents, the preovulatory surge in 17{beta}-estradiol (E2) temporarily allows energy expenditure to outpace energy intake, thus coordinating increased physical activity with peak sexual receptivity. To investigate how estrogen rebalances energy allocation in females, we examine estrogen receptor alpha (ER) signaling in the ventrolateral ventromedial hypothalamic nucleus (VMHvl)3-7. We uncover a small population of VMHvlER neurons expressing the melanocortin-4 receptor (MC4R) that integrates estrogen and melanocortin signals and projects to arousal centers in the hippocampus and hindbrain, enabling bursts of physical activity. ER recruitment to the Mc4r gene promotes upregulation of Mc4r in VMHvl neurons during the preovulatory surge or following E2 treatment. We leveraged three models to stimulate VMHvlMC4R neurons, restore MC4R signaling in the VMHvl of hyperphagic MC4R null females, or increase Mc4r levels in the VMHvl by CRISPR-mediated activation. All models increase spontaneous activity, whereas silencing VMHvlMC4R neurons blunts normal activity. Direct activation of the VMHvlMC4R node overrides the inactivity and hypometabolism following hormone depletion. These data extend the impact of MC4R signaling - the most common cause of monogenic human obesity8 - beyond the regulation of food intake. Our findings also rationalize reported sex differences in melanocortin signaling, including the greater disease severity of MC4R insufficiency in women9. The hormone-dependent node identified here illuminates the power of estrogen in motivating behavior during the female reproductive cycle and for maintaining an active lifestyle.
]]></description>
<dc:creator>Krause, W. C.</dc:creator>
<dc:creator>Rodriguez, R.</dc:creator>
<dc:creator>Gegenhuber, J. B.</dc:creator>
<dc:creator>Matharu, N.</dc:creator>
<dc:creator>Rodriguez, A. N.</dc:creator>
<dc:creator>Padilla, A.</dc:creator>
<dc:creator>Herber, C. B.</dc:creator>
<dc:creator>Correa, S. M.</dc:creator>
<dc:creator>Ahituv, N. M.</dc:creator>
<dc:creator>Tollkuhn, J.</dc:creator>
<dc:creator>Ingraham, H. A.</dc:creator>
<dc:date>2019-10-08</dc:date>
<dc:identifier>doi:10.1101/794792</dc:identifier>
<dc:title><![CDATA[Estrogen Drives Melanocortin Neurons To Reduce Sedentary Behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/794842v1?rss=1">
<title>
<![CDATA[
Broadly Conserved Roles of TMEM-131 Family Proteins in Intracellular Collagen Assembly and Secretory Cargo Trafficking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/794842v1?rss=1</link>
<description><![CDATA[
Collagen is the most abundant protein in animals. Its dysregulation contributes to ageing and human disorders including tissue fibrosis in major organs. How premature collagens in the endoplasmic reticulum (ER) assemble and route for secretion remains molecularly undefined. From an RNAi screen, we identified an uncharacterized C. elegans gene tmem-131, deficiency of which impairs collagen production and activates ER stress response. TMEM-131 N-termini contain bacterial PapD chaperone-like (PapD-L) domains essential for collagen assembly and secretion. Human TMEM131 binds to COL1A2 and TRAPPC8 via N-terminal PapD-L and C-terminal domain, respectively, to drive collagen production. We provide evidence that previously undescribed roles of TMEM131 in collagen recruitment and secretion are evolutionarily conserved in C. elegans, Drosophila and humans.
]]></description>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Bai, M.</dc:creator>
<dc:creator>Barbosa, G. O.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Luo, S.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Tsukui, T.</dc:creator>
<dc:creator>LI, H.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:creator>Kornberg, T.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2019-10-05</dc:date>
<dc:identifier>doi:10.1101/794842</dc:identifier>
<dc:title><![CDATA[Broadly Conserved Roles of TMEM-131 Family Proteins in Intracellular Collagen Assembly and Secretory Cargo Trafficking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/796912v1?rss=1">
<title>
<![CDATA[
Selectivity and engineering of the sialoglycan-binding spectrum in Siglec-like adhesins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/796912v1?rss=1</link>
<description><![CDATA[
The Siglec-like Serine-Rich Repeat (SRR) adhesins mediate bacterial attachment to mammalian hosts via sialoglycan receptors. Here, we combine structural, computational, biochemical, and phylogenetic approaches to elucidate the determinants of the sialoglycan-binding spectrum across the family of Siglec-like SRR adhesins. We further identified mutable positions that disproportionately affect sialoglycan selectivity, as demonstrated by increases in binding to alternative ligands of 2- to 3-orders of magnitude. Biologically, these studies highlight how bacteria nimbly modulate the receptor interaction during coevolution of host and pathogen. These studies additionally created binding proteins specific for sialyl-T antigen or 6S-sialyl LewisX that can recognize glycosylation of human plasma proteins. The engineered binding proteins can facilitate the characterization of normal cellular glycan modifications or may be used as diagnostic tools in disease states with altered glycosylation.nnSignificanceThe ability of bacteria to bind selectively to host receptors underlies both commensalism and pathogenesis. Here, we identify the molecular basis for receptor selectivity in streptococci that bind to sialoglycan receptors. This revealed how to convert these adhesins into selective probes that measure triand tetrasacharides within the context of larger glycosylations. These probes that can be used in a laboratory with no specialized equipment and can be used to address biological questions relating to sialoglycan-dependent signaling and adhesion.
]]></description>
<dc:creator>Bensing, B.</dc:creator>
<dc:creator>Loukachevitch, L. V.</dc:creator>
<dc:creator>Agarwal, R.</dc:creator>
<dc:creator>Yamakawa, I.</dc:creator>
<dc:creator>Luong, K.</dc:creator>
<dc:creator>Hadadianpour, A.</dc:creator>
<dc:creator>Yu, H.</dc:creator>
<dc:creator>Fialkowski, K.</dc:creator>
<dc:creator>Castro, M.</dc:creator>
<dc:creator>Wawrzak, Z.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Baudry, J.</dc:creator>
<dc:creator>Smith, J.</dc:creator>
<dc:creator>Sullam, P.</dc:creator>
<dc:creator>Iverson, T. M.</dc:creator>
<dc:date>2019-10-07</dc:date>
<dc:identifier>doi:10.1101/796912</dc:identifier>
<dc:title><![CDATA[Selectivity and engineering of the sialoglycan-binding spectrum in Siglec-like adhesins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/800060v1?rss=1">
<title>
<![CDATA[
Predicting 3D genome folding from DNA sequence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/800060v1?rss=1</link>
<description><![CDATA[
In interphase, the human genome sequence folds in three dimensions into a rich variety of locus-specific contact patterns. Here we present a deep convolutional neural network, Akita, that accurately predicts genome folding from DNA sequence alone. Representations learned by Akita underscore the importance of CTCF and reveal a complex grammar underlying genome folding. Akita enables rapid in silico predictions for sequence mutagenesis, genome folding across species, and genetic variants.
]]></description>
<dc:creator>Fudenberg, G.</dc:creator>
<dc:creator>Kelley, D. R.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2019-10-10</dc:date>
<dc:identifier>doi:10.1101/800060</dc:identifier>
<dc:title><![CDATA[Predicting 3D genome folding from DNA sequence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/803288v1?rss=1">
<title>
<![CDATA[
Paralytic, the Drosophila voltage-gated sodium channel, regulates proliferation of neural progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/803288v1?rss=1</link>
<description><![CDATA[
Proliferating cells, typically considered "non-excitable," nevertheless exhibit regulation by bioelectrical signals. Notably, voltage-gated sodium channels (VGSC) that are crucial for neuronal excitability, are also found in progenitors and upregulated in cancer. Here, we identify a role for VGSC in proliferation of Drosophila neuroblast (NB) lineages within the central nervous system. Loss of paralytic (para), the sole gene that encodes Drosophila VGSC, reduces neuroblast progeny cell number. The type II neuroblast lineages, featuring a transit-amplifying intermediate neural progenitors (INP) population similar to that found in the developing human cortex, are particularly sensitive to para manipulation. Following a series of asymmetric divisions, INPs normally exit the cell cycle through a final symmetric division. Our data suggests that loss of para induces apoptosis in this population, whereas overexpression leads to an increase in INPs and overall neuroblast progeny cell numbers. These effects are cell autonomous and depend on Para channel activity. Reduction of Para not only affects normal NB development, but also strongly suppresses brain tumor mass, implicating a role for Para in cancer progression. To our knowledge, our studies are the first to identify a role for VGSC in neural progenitor proliferation. Elucidating the contribution of VGSC in proliferation will advance our understanding of bioelectric signaling within development and disease states.
]]></description>
<dc:creator>Piggott, B. J.</dc:creator>
<dc:creator>Peters, C. J.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Younger, S.</dc:creator>
<dc:creator>Jan, L.</dc:creator>
<dc:creator>Jan, Y. N.</dc:creator>
<dc:date>2019-10-14</dc:date>
<dc:identifier>doi:10.1101/803288</dc:identifier>
<dc:title><![CDATA[Paralytic, the Drosophila voltage-gated sodium channel, regulates proliferation of neural progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/805929v1?rss=1">
<title>
<![CDATA[
Dopamine and risky decision-making in pathological and problem gamblers. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/805929v1?rss=1</link>
<description><![CDATA[
Gambling disorder is a behavioral addiction that is associated with impairments in value-based decision-making such as increased temporal discounting and reduced risk-aversion. Dopamine regulates learning and decision-making by modulating information processing throughout fronto-striatal circuits. Although the role of alterations in dopamine neurotransmission in the etiology of gambling disorder is controversial, preliminary evidence suggests that specifically increasing frontal dopamine levels might improve cognitive functioning in pathological and problem gamblers. We therefore examined whether increasing frontal dopamine levels via the catechol-O-methyltransferase (COMT) inhibitor tolcapone would reduce risky choice in a group of pathological and problem gamblers (n=14) in a repeated-measures counter-balanced placebo-controlled double-blind study. Choice data were fit using hierarchical Bayesian parameter estimation and a modeling scheme that combined a risky choice model with the drift diffusion model to account for both choices and response time distributions. Model comparison revealed that the data were best accounted for by a variant of the drift diffusion model with a non-linear modulation of trial-wise drift rates by value differences, confirming recent findings. Contrary to our hypothesis, risk-taking was slightly increased under tolcapone vs. placebo (Cohens d = -.281). Examination of drug effects on diffusion model parameters revealed an increase in the value-dependency of the drift rate (Cohens d = .932) with a simultaneous reduction in the maximum drift rate (Cohens d = -1.84). These results add to previous work on the potential role of COMT inhibitors in behavioral addictions, and show no consistent beneficial effect of tolcapone on risky choice in gambling disorder. Modeling results add to mounting evidence for the applicability of diffusion models in value-based decision-making. Future work should focus on individual genetic, clinical and cognitive factors that might account for the heterogeneity in the effects of COMT inhibition.
]]></description>
<dc:creator>Peters, J.</dc:creator>
<dc:creator>Vega, T.</dc:creator>
<dc:creator>Weinstein, D.</dc:creator>
<dc:creator>Mitchell, J.</dc:creator>
<dc:creator>Kayser, A.</dc:creator>
<dc:date>2019-10-16</dc:date>
<dc:identifier>doi:10.1101/805929</dc:identifier>
<dc:title><![CDATA[Dopamine and risky decision-making in pathological and problem gamblers.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/807164v1?rss=1">
<title>
<![CDATA[
High throughput CRISPR screening identifies genes involved in macrophage viability and inflammatory pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/807164v1?rss=1</link>
<description><![CDATA[
AbstractMacrophages are critical cells of the innate immune system involved in the recognition and destruction of invading microbes in addition to the resolution of inflammation and maintenance of homeostasis. Understanding the genes involved in all aspects of macrophage biology is essential to gaining new insights into immune system dysregulation during diseases that range from autoinflammatory to cancer. Here we utilize high throughput clustered regularly interspaced short palindromic repeats (CRISPR) screening to generate a resource that identifies genes required for macrophage viability and function. First, we employ a pooled based CRISPR/Cas nuclease active screening approach to identify essential genes required for macrophage viability by targeting genes within coding exons. In addition, we also target 3UTRs to gain insights into new cis-regulatory regions that control expression of these essential genes. Second, using our recently generated NF-{kappa}B reporter macrophage line, we perform a fluorescence-activated cell sorting (FACS)-based high-throughput genetic screen to identify regulators of inflammation. We identify a number of novel positive and negative regulators of the NF-{kappa}B pathway as well as unraveling complexities of the TNF signaling cascade showing it can function in an autocrine manner to negatively regulate the pathway. Utilizing a single complex library design we are capable of interrogating various aspects of macrophage biology, generating a resource for future studies.nnAuthor SummaryExcess inflammation is associated with a variety of autoimmune diseases and cancers. Macrophages are important mediators of this inflammatory response. Defining the genes involved in their viability and effector function is needed to completely understand these two important aspects of macrophage biology. Here we screened over 21,000 genes and generated a resource guide of genes required for macrophage viability as well as novel positive and negative regulators of NF-{kappa}B signaling.
]]></description>
<dc:creator>Covarrubias, S.</dc:creator>
<dc:creator>Vollmers, A.</dc:creator>
<dc:creator>Capili, A.</dc:creator>
<dc:creator>Boettcher, M.</dc:creator>
<dc:creator>Robinson, E. K.</dc:creator>
<dc:creator>O'Briain, L.</dc:creator>
<dc:creator>Vollmers, C.</dc:creator>
<dc:creator>Blau, J.</dc:creator>
<dc:creator>McManus, M.</dc:creator>
<dc:creator>Carpenter, S.</dc:creator>
<dc:date>2019-10-16</dc:date>
<dc:identifier>doi:10.1101/807164</dc:identifier>
<dc:title><![CDATA[High throughput CRISPR screening identifies genes involved in macrophage viability and inflammatory pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/807297v1?rss=1">
<title>
<![CDATA[
Opioids depress breathing through two small brainstem sites 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/807297v1?rss=1</link>
<description><![CDATA[
The rates of opioid overdose in the United States quadrupled between 1999 and 2017, reaching a staggering 130 deaths per day. This health epidemic demands innovative solutions that require uncovering the key brain areas and cell types mediating the cause of overdose--opioid respiratory depression. Here, we identify two primary changes to breathing after administering opioids. These changes implicate the brainstems breathing circuitry which we confirm by locally eliminating the -Opiate receptor. We find the critical brain site is the origin of the breathing rhythm, the preBotzinger Complex, and use genetic tools to reveal that just 70-140 neurons in this region are responsible for its sensitivity to opioids. Future characterization of these neurons may lead to novel therapies that prevent respiratory depression while sparing analgesia.
]]></description>
<dc:creator>Iris, B.</dc:creator>
<dc:creator>Wei, X. P.</dc:creator>
<dc:creator>Kish, E.</dc:creator>
<dc:creator>Yackle, K.</dc:creator>
<dc:date>2019-10-17</dc:date>
<dc:identifier>doi:10.1101/807297</dc:identifier>
<dc:title><![CDATA[Opioids depress breathing through two small brainstem sites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/808006v1?rss=1">
<title>
<![CDATA[
Dynamic preferences account for inter-animal variability during the continual learning of a cognitive task 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/808006v1?rss=1</link>
<description><![CDATA[
Individual animals perform tasks in different ways, yet the nature and origin of that variability is poorly understood. In the context of spatial memory tasks, variability is often interpreted as resulting from differences in memory ability, but the validity of this interpretation is seldom tested since we lack a systematic approach for identifying and understanding factors that make one animals behavior different than another. Here we identify such factors in the context of spatial alternation in rats, a task often described as relying solely on memory of past choices. We combine hypothesis-driven behavioral design and reinforcement learning modeling to identify spatial preferences that, when combined with memory, support learning of a spatial alternation task. Identifying these preferences allows us to capture differences among animals, including differences in overall learning ability. Our results show that to understand the complexity of behavior requires quantitative accounts of the preferences of each animal.
]]></description>
<dc:creator>Kastner, D. B.</dc:creator>
<dc:creator>Miller, E. A.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Roumis, D. K.</dc:creator>
<dc:creator>Liu, D. F.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:creator>Dayan, P.</dc:creator>
<dc:date>2019-10-17</dc:date>
<dc:identifier>doi:10.1101/808006</dc:identifier>
<dc:title><![CDATA[Dynamic preferences account for inter-animal variability during the continual learning of a cognitive task]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/808139v1?rss=1">
<title>
<![CDATA[
Broad and thematic remodeling of the surface glycoproteome on isogenic cells transformed with driving proliferative oncogenes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/808139v1?rss=1</link>
<description><![CDATA[
The cell surface proteome, the surfaceome, is the interface for engaging the extracellular space in normal and cancer cells. Here we apply quantitative proteomics of N-linked glycoproteins to reveal how a collection of some 700 surface proteins is dramatically remodeled in an isogenic breast epithelial cell line stably expressing any of six of the most prominent proliferative oncogenes, including the receptor tyrosine kinases, EGFR and HER2, and downstream signaling partners such as KRAS, BRAF, MEK and AKT. We find that each oncogene has somewhat different surfaceomes but the functions of these proteins are harmonized by common biological themes including up-regulation of nutrient transporters, down-regulation of adhesion molecules and tumor suppressing phosphatases, and alteration in immune modulators. Addition of a potent MEK inhibitor that blocks MAPK signaling brings each oncogene-induced surfaceome back to a common state reflecting their strong dependence on the MAPK pathway to propagate signaling. Using a recently developed glyco-proteomics method of activated ion electron transfer dissociation (AI-ETD) we found massive oncogene-induced changes in 142 N-linked glycans and differential increases in complex hybrid glycans especially for KRAS and HER2 oncogenes. Overall, these studies provide a broad systems level view of how specific driver oncogenes remodel the surface glycoproteome in a cell autologous fashion, and suggest possible surface targets, and combinations thereof, for drug and biomarker discovery.nnSignificant statementThe cell surface glycoproteome (surfaceome) mediates interactions between the cell and the extracellular environment, and is a major target for immunotherapy in cancer. Using state-of-the-art proteomics methods, we compared how six neighboring proliferative oncogenes cause large and bidirectional expression of some 700 surface proteins and the 142 different glycans that decorate them. While each oncogene induces large and somewhat unique glycoproteomes relative to non-transformed cells, we find common functional consequences that are massively reversed by small molecule inhibition of the MAPK pathway. This large-scale comparative study provides important insights for how oncogenes remodel isogenic cells in a cell autologous fashion, and suggest possible new opportunities for antibody drug discovery in more complex tumor settings.
]]></description>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Wilson, G. M.</dc:creator>
<dc:creator>Kirkemo, L. L.</dc:creator>
<dc:creator>Riley, N. M.</dc:creator>
<dc:creator>Coon, J. J.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2019-10-17</dc:date>
<dc:identifier>doi:10.1101/808139</dc:identifier>
<dc:title><![CDATA[Broad and thematic remodeling of the surface glycoproteome on isogenic cells transformed with driving proliferative oncogenes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/810721v1?rss=1">
<title>
<![CDATA[
EGFR Oncogenes Expressed In Glioblastoma Are Activated As Covalent Dimers And Paradoxically Stimulated By Erlotinib 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/810721v1?rss=1</link>
<description><![CDATA[
Mutation of both the intracellular catalytic domain and the extracellular domain of the receptor for epidermal growth factor (EGFR) can drive oncogenicity. Despite clinical success with targeting EGFR catalytic site mutations, no drugs have proven effective in patients expressing allosteric extracellular domain EGFR mutations, including glioblastomas (GBM) where these mutations are highly expressed. We define the molecular mechanism for oncogenic activation of families of extracellular EGFR mutations and reveal how this mechanism renders current generation small molecule ATP-site inhibitors ineffective. We demonstrate that a group of commonly expressed extracellular domain EGFR mutants expressed in GBM is activated by disulfide-bond mediated covalent dimerization, collectively referred to as locked dimerization (LoDi) EGFR oncogenes. Strikingly, small molecules binding to the active kinase conformation (Type I), but not those binding to the inactive kinase conformation (Type II), potently inhibit catalytic site mutants, but induce covalent dimerization and activate LoDi-EGFR oncogenes, manifesting in paradoxical acceleration of proliferation.nnSignificanceOur data demonstrate how the locked-dimer mechanism of EGFR oncogenesis has a profound impact on the activity of small molecule inhibitors. This provides a mechanistic understanding for the failure of current generation EGFR inhibitors to effectively treat LoDi-EGFR mutants in GBM, and sets guidelines for discovery of selective LoDi-EGFR inhibitors.
]]></description>
<dc:creator>Buck, E.</dc:creator>
<dc:creator>O'Connor, M.</dc:creator>
<dc:creator>Romashko, D.</dc:creator>
<dc:creator>Salomatov, A.</dc:creator>
<dc:creator>Ishiyama, N.</dc:creator>
<dc:creator>Epstein, D.</dc:creator>
<dc:creator>Mayweg, A.</dc:creator>
<dc:creator>Flohr, A.</dc:creator>
<dc:creator>Iacone, R.</dc:creator>
<dc:creator>Nicolaides, T.</dc:creator>
<dc:creator>Markovic, S.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:date>2019-10-21</dc:date>
<dc:identifier>doi:10.1101/810721</dc:identifier>
<dc:title><![CDATA[EGFR Oncogenes Expressed In Glioblastoma Are Activated As Covalent Dimers And Paradoxically Stimulated By Erlotinib]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/811083v1?rss=1">
<title>
<![CDATA[
Liquid-like and rigid-body motions in molecular-dynamics simulations of a crystalline protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/811083v1?rss=1</link>
<description><![CDATA[
To gain insight into crystalline protein dynamics, we performed molecular-dynamics (MD) simulations of a periodic 2x2x2 supercell of staphylococcal nuclease. We used the resulting MD trajectories to simulate X-ray diffraction and to study collective motions. The agreement of simulated X-ray diffraction with the data is comparable to previous MD simulation studies. We studied collective motions by analyzing statistically the covariance of alpha-carbon position displacements. The covariance decreases exponentially with the distance between atoms, which is consistent with a liquid-like motions (LLM) model, in which the protein behaves like a soft material. To gain finer insight into the collective motions, we examined the covariance behavior within a protein molecule (intra-protein) and between different protein molecules (inter-protein). The inter-protein atom pairs, which dominate the overall statistics, exhibit LLM behavior; however, the intra-protein pairs exhibit behavior that is consistent with a superposition of LLM and rigid-body motions (RBM). Our results indicate that LLM behavior of global dynamics is present in MD simulations of a protein crystal. They also show that RBM behavior is detectable in the simulations but that it is subsumed by the LLM behavior. Finally the results provide clues about how correlated motions of atom pairs both within and across proteins might manifest in diffraction data. Overall our findings increase our understanding of the connection between molecular motions and diffraction data, and therefore advance efforts to extract information about functionally important motions from crystallography experiments.
]]></description>
<dc:creator>Wych, D. C.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Mobley, D. L.</dc:creator>
<dc:creator>Wall, M.</dc:creator>
<dc:date>2019-10-18</dc:date>
<dc:identifier>doi:10.1101/811083</dc:identifier>
<dc:title><![CDATA[Liquid-like and rigid-body motions in molecular-dynamics simulations of a crystalline protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/811620v1?rss=1">
<title>
<![CDATA[
A Chromatin Accessibility Atlas of the Developing Human Telencephalon 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/811620v1?rss=1</link>
<description><![CDATA[
Gene expression differs between cell types and regions within complex tissues such as the developing brain. To discover regulatory elements underlying this specificity, we generated genome-wide maps of chromatin accessibility in eleven anatomically-defined regions of the developing human telencephalon, including upper and deep layers of the prefrontal cortex. We predicted a subset of open chromatin regions (18%) that are most likely to be active enhancers, many of which are dynamic with 26% differing between early and late mid-gestation and 28% present in only one brain region. These region-specific predicted regulatory elements (pREs) are enriched proximal to genes with expression differences across regions and developmental stages and harbor distinct sequence motifs that suggest potential upstream regulators of regional and temporal transcription. We leverage this atlas to identify regulators of genes associated with autism spectrum disorder (ASD) including an enhancer of BCL11A, validated in mouse, and two functional de novo mutations in individuals with ASD in an enhancer of SLC6A1, validated in neuroblastoma cells. These applications demonstrate the utility of this atlas for decoding neurodevelopmental gene regulation in health and disease.nnSummaryTo discover regulatory elements driving the specificity of gene expression in different cell types and regions of the developing human brain, we generated an atlas of open chromatin from eleven dissected regions of the mid-gestation human telencephalon, including upper and deep layers of the prefrontal cortex. We identified a subset of open chromatin regions (OCRs), termed predicted regulatory elements (pREs), that are likely to function as developmental brain enhancers. pREs showed regional differences in chromatin accessibility, including many specific to one brain region, and were correlated with gene expression differences across the same regions and gestational ages. pREs allowed us to map neurodevelopmental disorder risk genes to developing telencephalic regions, and we identified three functional de novo noncoding variants in pREs that alter enhancer function. In addition, transgenic experiments in mouse validated enhancer activity for a pRE proximal to BCL11A, showing how this atlas serves as a resource for decoding neurodevelopmental gene regulation in health and disease.
]]></description>
<dc:creator>Markenscoff-Papadimitriou, E.</dc:creator>
<dc:creator>Whalen, S.</dc:creator>
<dc:creator>Przytycki, P.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Binyameen, F.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Rubenstein, J. L.</dc:creator>
<dc:date>2019-10-21</dc:date>
<dc:identifier>doi:10.1101/811620</dc:identifier>
<dc:title><![CDATA[A Chromatin Accessibility Atlas of the Developing Human Telencephalon]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/813972v1?rss=1">
<title>
<![CDATA[
Amino and PEG-Amino Graphene Oxide Grids Enrich and Protect Samples for High-resolution Single Particle Cryo-electron Microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/813972v1?rss=1</link>
<description><![CDATA[
Cryo-EM samples prepared using the traditional methods often suffer from too few particles, poor particle distribution, or strongly biased orientation, or damage from the air-water interface. Here we report that functionalization of graphene oxide (GO) coated grids with amino groups concentrates samples on the grid with improved distribution and orientation. By introducing a PEG spacer, particles are kept away from both the GO surface and the air-water interface, protecting them from potential denaturation.
]]></description>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Betegon, M.</dc:creator>
<dc:creator>Campbell, M.</dc:creator>
<dc:creator>Aksel, T.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Douglas, S. M.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Agard, D.</dc:creator>
<dc:date>2019-10-22</dc:date>
<dc:identifier>doi:10.1101/813972</dc:identifier>
<dc:title><![CDATA[Amino and PEG-Amino Graphene Oxide Grids Enrich and Protect Samples for High-resolution Single Particle Cryo-electron Microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/814087v1?rss=1">
<title>
<![CDATA[
Genetic and clinical analyses of psychosis spectrum symptoms in a large multi-ethnic youth cohort reveal significant link with ADHD. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/814087v1?rss=1</link>
<description><![CDATA[
ObjectivePsychotic symptoms are an important feature of severe neuropsychiatric disorders, but are also common in the general population, especially in youth. The genetic etiology of psychosis symptoms in youth remains poorly understood. To characterize genetic risk for psychosis spectrum symptoms (PS), we leverage a community-based multi-ethnic sample of children and adolescents aged 8-22 years, the Philadelphia Neurodevelopmental Cohort (n = 7,225, 20% PS).nnMethodsUsing an elastic net regression model, we aim to classify PS status using polygenic scores (PGS) based on a range of heritable psychiatric and brain-related traits in a multi-PGS model. We also perform univariate PGS associations and evaluate age-specific effects.nnResultsThe multi-PGS analyses do not improve prediction of PS status over univariate models, but reveal that the attention deficit hyperactivity disorder (ADHD) PGS is robustly and uniquely associated with PS (OR 1.12 (1.05, 1.18) P = 0.0003). This association is: i) driven by subjects of European ancestry (OR=1.23 (1.14, 1.34), P=4.15x10-7) but is not observed in African American subjects (P=0.65) and ii) independent of phenotypic overlap. We also find a significant interaction with age (P=0.01), with a stronger association in younger children. In an independent sample, we replicate an increased ADHD PGS in 328 youth at clinical high risk for psychosis, compared to 216 unaffected controls (OR 1.06, CI(1.01, 1.11), P= 0.02).nnConclusionsOur findings suggest that PS in youth may reflect a different genetic etiology than psychotic symptoms in adulthood, one more akin to ADHD, and shed light on how genetic risk can be investigated across early disease trajectories.
]]></description>
<dc:creator>Olde Loohuis, L. M.</dc:creator>
<dc:creator>Mennigen, E.</dc:creator>
<dc:creator>Ori, A. P. S.</dc:creator>
<dc:creator>Perkins, D.</dc:creator>
<dc:creator>Robinson, E.</dc:creator>
<dc:creator>Addington, J.</dc:creator>
<dc:creator>Cadenhead, K. S.</dc:creator>
<dc:creator>Cornblatt, B. A.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>McGlashan, T. H.</dc:creator>
<dc:creator>Seidman, L. J.</dc:creator>
<dc:creator>Keshavan, M.</dc:creator>
<dc:creator>Stone, W.</dc:creator>
<dc:creator>Tsuang, M. T.</dc:creator>
<dc:creator>Walker, E. F.</dc:creator>
<dc:creator>Woods, S. W.</dc:creator>
<dc:creator>Cannon, T. D.</dc:creator>
<dc:creator>Gur, R. C.</dc:creator>
<dc:creator>Gur, R. E.</dc:creator>
<dc:creator>Bearden, C. E.</dc:creator>
<dc:creator>Ophoff, R. A.</dc:creator>
<dc:date>2019-10-22</dc:date>
<dc:identifier>doi:10.1101/814087</dc:identifier>
<dc:title><![CDATA[Genetic and clinical analyses of psychosis spectrum symptoms in a large multi-ethnic youth cohort reveal significant link with ADHD.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/815811v1?rss=1">
<title>
<![CDATA[
Phenotype-based screening of synthetic cannabinoids in a Dravet Syndrome zebrafish model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/815811v1?rss=1</link>
<description><![CDATA[
Dravet syndrome (DS) is a catastrophic epilepsy of childhood, characterized by cognitive impairment, severe seizures and increased risk for sudden unexplained death in epilepsy (SUDEP). Although refractory to conventional antiepileptic drugs, emerging preclinical and clinical evidence suggests that modulation of the endocanniboid system could be therapeutic in these patients. Here we used a validated zebrafish model of DS, scn1lab homozygous mutants, to screen a commercially available library containing 370 synthetic cannabinoid (SC) compounds for compounds effective in reducing spontaneous seizures. Primary phenotype-based screening was performed using a locomotion-based assay in 96-well plates, and a secondary local field potential recording assay was then used to confirm suppression of electrographic epileptiform events. Identified SCs with anti-seizure activity, in both assays, included five SCs structurally classified as indole-based cannabinoids: JWH 018 N-(5-chloropentyl) analog, JWH 018 N-(2-methylbutyl) isomer, 5-fluoro PB-22 5-hydroxyisoquinoline isomer, 5-fluoro ADBICA, and AB-FUBINACA 3-fluorobenzyl isomer. Our approach demonstrates that two-stage phenotype-based screening in a zebrafish model of DS successfully identifies synthetic cannabinoids with anti-seizure activity, and supports further investigation of SCs for refractory epilepsies.
]]></description>
<dc:creator>Griffin, A.</dc:creator>
<dc:creator>Anvar, M.</dc:creator>
<dc:creator>Hamling, K.</dc:creator>
<dc:creator>Baraban, S. C.</dc:creator>
<dc:date>2019-10-23</dc:date>
<dc:identifier>doi:10.1101/815811</dc:identifier>
<dc:title><![CDATA[Phenotype-based screening of synthetic cannabinoids in a Dravet Syndrome zebrafish model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/820084v1?rss=1">
<title>
<![CDATA[
Drug resistant gut bacteria mimic a host mechanism for anticancer drug clearance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/820084v1?rss=1</link>
<description><![CDATA[
Pharmaceuticals are the top predictor of inter-individual variations in gut microbial community structure1, consistent with in vitro evidence that host-targeted drugs inhibit gut bacterial growth2 and are extensively metabolized by the gut microbiome3,4. In oncology, bacterial metabolism has been implicated in both drug efficacy5,6 and toxicity7,8; however, the degree to which bacterial drug sensitivity and metabolism can be driven by conserved pathways also found in mammalian cells remains poorly understood. Here, we show that anticancer fluoropyrimidine drugs inhibit the growth of diverse gut bacterial strains by disrupting pyrimidine metabolism, as in mammalian cells. Select bacteria metabolized 5-fluorouracil (5-FU) to its inactive metabolite dihydrofluorouracil (DHFU), mimicking the major host pathway for drug clearance. The preTA operon was necessary and sufficient for 5-FU inactivation in Escherichia coli, exhibited high catalytic efficiency for the reductive reaction, decreased the bioavailability and efficacy of oral fluoropyrimidine treatment in mice, and was prevalent in the gut microbiomes of colorectal cancer patients prior to and during treatment. The observed conservation of both the targets and pathways for metabolism of therapeutics across domains highlights the need to distinguish the relative contributions of human and microbial cells to drug disposition9, efficacy, and side effect profiles.
]]></description>
<dc:creator>Spanogiannopoulos, P.</dc:creator>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:creator>Melamed, J.</dc:creator>
<dc:creator>Malig, Y. N. A.</dc:creator>
<dc:creator>Lam, K. N.</dc:creator>
<dc:creator>Gerona, R. R.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Turnbaugh, P.</dc:creator>
<dc:date>2019-10-25</dc:date>
<dc:identifier>doi:10.1101/820084</dc:identifier>
<dc:title><![CDATA[Drug resistant gut bacteria mimic a host mechanism for anticancer drug clearance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/825836v1?rss=1">
<title>
<![CDATA[
Obesity Potentiates TH2 Immunopathology via Dysregulation of PPAR 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/825836v1?rss=1</link>
<description><![CDATA[
How obesity affects immune function is not well understood. Clinically, obesity is strongly associated with severe TH2 immunopathology1-3, though the physiological, cellular, and molecular underpinnings of this association remain obscure. Here, we demonstrate that obese mice are susceptible to severe atopic dermatitis (AD), a major manifestation of TH2 immunopathology and disease burden in humans4,5. Mechanistically, we show that dysregulation of the nuclear hormone receptor (NHR) PPAR{gamma} (peroxisome proliferator-activated receptor gamma) in T cells is a causal link between obesity and the increased TH2 immunopathology. We find that PPAR{gamma} oversees a cellular metabolic transcriptional program that restrains nuclear gene expression of the chief TH2 priming and effector cytokine interleukin-4 (IL-4). Accordingly, thiazolidinediones (TZDs), potent PPAR{gamma} agonists, robustly protect obese mice from TH2 immunopathology. Collectively, these findings establish PPAR{gamma} as a molecular link between obesity and TH2 immune homeostasis and identify TZDs as novel therapeutic candidates for TH2 immunopathology. Fundamentally, these findings demonstrate that shifting physiologic metabolic states can shape the tone of adaptive immune responses to modulate differential disease susceptibility.
]]></description>
<dc:creator>Bapat, S.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Zhang, L.-j.</dc:creator>
<dc:creator>Vogel, I.</dc:creator>
<dc:creator>Marr, D. J.</dc:creator>
<dc:creator>Zhou, C.</dc:creator>
<dc:creator>Choi, E. J.</dc:creator>
<dc:creator>Chang, C.</dc:creator>
<dc:creator>He, N.</dc:creator>
<dc:creator>LeBlanc, M.</dc:creator>
<dc:creator>Lee, I.-k.</dc:creator>
<dc:creator>Suh, J. M.</dc:creator>
<dc:creator>Crotty Alexander, L. E.</dc:creator>
<dc:creator>Liddle, C.</dc:creator>
<dc:creator>Atkins, A. R.</dc:creator>
<dc:creator>Yu, R. T.</dc:creator>
<dc:creator>Downes, M.</dc:creator>
<dc:creator>Ansel, K. M.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Gallo, R. L.</dc:creator>
<dc:creator>Evans, R. M.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:date>2019-10-31</dc:date>
<dc:identifier>doi:10.1101/825836</dc:identifier>
<dc:title><![CDATA[Obesity Potentiates TH2 Immunopathology via Dysregulation of PPAR]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/829465v1?rss=1">
<title>
<![CDATA[
Recurrent loss of abaA, a master regulator of asexual development in filamentous fungi, correlates with changes in genomic and morphological traits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/829465v1?rss=1</link>
<description><![CDATA[
Gene regulatory networks (GRNs) drive developmental and cellular differentiation, and variation in their architectures gives rise to morphological diversity. Pioneering studies in Aspergillus fungi, coupled with subsequent work in other filamentous fungi, have shown that the GRN governed by the BrlA, AbaA, and WetA proteins controls the development of the asexual fruiting body or conidiophore. A specific aspect of conidiophore development is the production of phialides, conidiophore structures that are under the developmental control of AbaA and function to repetitively generate spores. Fungal genome sequencing has revealed that some filamentous fungi lack abaA, and also produce asexual structures that lack phialides, raising the hypothesis that abaA loss is functionally linked to diversity in asexual fruiting body morphology. To examine this hypothesis, we carried out an extensive search for the abaA gene across 241 genomes of species from the fungal subphylum Pezizomycotina. We found that abaA was independently lost in four lineages of Eurotiomycetes, including from all sequenced species within the order Onygenales, and that all four lineages that have lost abaA also lack the ability to form phialides. Genetic restoration of abaA from Aspergillus nidulans into Histoplasma capsulatum, a pathogenic species from the order Onygenales that lacks an endogenous copy of abaA, did not alter Histoplasma conidiation morphology but resulted in a marked increase in spore viability. We also discovered that species lacking abaA contain fewer AbaA binding motifs in the regulatory regions of orthologs of some AbaA target genes, suggesting that the asexual fruiting body GRN of organisms that have lost abaA has been rewired. Our results provide an illustration of how repeated losses of a key regulatory transcription factor and concomitant changes in non-coding regulatory regions of the genome have contributed to the diversity of an iconic fungal morphological trait.nnAuthor SummaryFungi exhibit tremendous variation in their asexual fruiting bodies. For example, whereas some fungi form complex fruiting bodies whose tips repeatedly generate and release spores, others produce single spores in the absence of a specialized structure. To gain insights into the molecular differences that underpin fungal asexual fruiting body diversity, we examined the genomes of hundreds of filamentous fungi for the presence of abaA, a master regulatory gene previously shown to control the development of fungal asexual fruiting bodies. We found that abaA was repeatedly lost during fungal evolution, including in a lineage of human pathogenic fungi, and that the loss of the gene was always associated with the loss of specialized structures in fungal asexual fruiting bodies. Reintroduction of abaA into the human pathogenic fungus Histoplasma capsulatum, which normally lacks the regulator, did not result in a change in the spore-producing structure but did increase spore outgrowth. Based on these results, we hypothesize that the loss of the master regulatory gene abaA has contributed to the observed diversity of fungal fruiting body morphology. This work advances our understanding of how fungal developmental networks evolve over time and advances our understanding of how infectious spores form in pathogenic fungi.
]]></description>
<dc:creator>Mead, M. E.</dc:creator>
<dc:creator>Borowsky, A. T.</dc:creator>
<dc:creator>Joehnk, B.</dc:creator>
<dc:creator>Steenwyk, J. L.</dc:creator>
<dc:creator>Shen, X.-X.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:creator>Rokas, A.</dc:creator>
<dc:date>2019-11-04</dc:date>
<dc:identifier>doi:10.1101/829465</dc:identifier>
<dc:title><![CDATA[Recurrent loss of abaA, a master regulator of asexual development in filamentous fungi, correlates with changes in genomic and morphological traits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/833186v1?rss=1">
<title>
<![CDATA[
PCK1 and DHODH drive colorectal cancer liver metastatic colonization and hypoxic pyrimidine nucleotide anabolic biosynthesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/833186v1?rss=1</link>
<description><![CDATA[
Colorectal cancer (CRC) is a major cause of human death. Mortality is primarily due to metastatic organ colonization, with liver being the primary organ affected. We modeled metastatic CRC (mCRC) liver colonization using patient-derived primary and metastatic tumor xenografts (PDX). Such PDX modeling predicted patient survival outcomes. In vivo selection of multiple PDXs for enhanced metastatic capacity upregulated the gluconeogenic enzyme PCK1, which enhanced metastatic hypoxic survival by driving anabolic pyrimidine nucleotide biosynthesis. Consistently, highly metastatic tumors upregulated multiple pyrimidine biosynthesis intermediary metabolites. Therapeutic inhibition of the pyrimidine biosynthetic enzyme DHODH with oral leflunomide substantially impaired CRC liver metastatic colonization and hypoxic survival. Our findings provide a potential mechanistic basis for the epidemiologic association of anti-gluconeogenic drugs with improved CRC metastasis outcomes, reveal the exploitation of a gluconeogenesis enzyme for pyrimidine biosynthesis during hypoxia, and implicate DHODH and PCK1 as metabolic therapeutic targets in colorectal cancer metastasis.
]]></description>
<dc:creator>Yamaguchi, N.</dc:creator>
<dc:creator>Weinberg, E.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Liberti, M. V.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Janjigian, Y. Y.</dc:creator>
<dc:creator>Paty, P. B.</dc:creator>
<dc:creator>Saltz, L. B.</dc:creator>
<dc:creator>Kingham, T. P.</dc:creator>
<dc:creator>Loo, J. M.</dc:creator>
<dc:creator>Stanchina, E. d.</dc:creator>
<dc:creator>Tavazoie, S. F.</dc:creator>
<dc:date>2019-11-06</dc:date>
<dc:identifier>doi:10.1101/833186</dc:identifier>
<dc:title><![CDATA[PCK1 and DHODH drive colorectal cancer liver metastatic colonization and hypoxic pyrimidine nucleotide anabolic biosynthesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/840306v1?rss=1">
<title>
<![CDATA[
α-synuclein aggregates induce c-Abl activation and neurodegeneration by a redox stress mechanism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/840306v1?rss=1</link>
<description><![CDATA[
ObjectiveRedox stress, c-Abl activation, and -synuclein aggregates each independently contribute to neurodegeneration in Parkinsons disease. Interactions between these factors may underlie convergent and feed-forward mechanisms of disease progression.

Methods-synuclein aggregate formation was induced in neuronal cultures and mouse substantia nigra by exposure to pre-formed human -synuclein fibrils or by AAV-mediated over-expression of -synuclein. Aggregate formation, c-Abl activation, redox stress, and neurodegeneration were evaluated by immunohistochemistry and Western blots, and mouse motor function was evaluated using the rota-rod and pole tests. To suppress redox stress, cultures and mice were treated with N-acetyl cysteine, a thiol repletion agent that supports neuronal glutathione metabolism.

ResultsIn primary neuron cultures, the formation of -synuclein aggregates led to redox stress and c-Abl activation. Redox stress alone, in the absence of -synuclein aggregates, was also sufficient to induced c-Abl activation. N-acetyl cysteine suppressed redox stress, and likewise suppressed both c-Abl activation and -synuclein aggregation. A similar pattern was observed in the two mouse models of Parkinsons disease. In both models, -synuclein aggregates in the substantia nigra were accompanied by redox stress, c-Abl activation, dopaminergic neurodegeneration and motor impairment, all of which were attenuated in mice treated with oral N-acetyl cysteine.

InterpretationThese results indicate that -synuclein aggregates induce c-Abl activation by a redox stress mechanism. c-Abl in turn promotes -synuclein aggregation, and this potentially feed-forward process can be blocked by N-acetyl cysteine. The findings thus add mechanistic support for N-acetyl cysteine as a therapeutic for Parkinsons disease.
]]></description>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Won, S. J.</dc:creator>
<dc:creator>Fong, R.</dc:creator>
<dc:creator>Butler, N. J. M.</dc:creator>
<dc:creator>Moss, A.</dc:creator>
<dc:creator>Wong, C.</dc:creator>
<dc:creator>Pan, J.</dc:creator>
<dc:creator>Sanchez, J.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Manfredsson, F. P.</dc:creator>
<dc:creator>Swanson, R. A.</dc:creator>
<dc:date>2019-11-13</dc:date>
<dc:identifier>doi:10.1101/840306</dc:identifier>
<dc:title><![CDATA[α-synuclein aggregates induce c-Abl activation and neurodegeneration by a redox stress mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/841932v1?rss=1">
<title>
<![CDATA[
Combined modulation of SHH and FGF signaling is crucial for maintenance of the neocortical progenitor specification program 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/841932v1?rss=1</link>
<description><![CDATA[
Neuronal progenitors in the developing forebrain undergo dynamic competence states to ensure timely generation of specific excitatory and inhibitory neuronal subtypes from distinct neurogenic niches of the dorsal and ventral forebrain, respectively. Here we show evidence of progenitor plasticity when Sonic hedgehog (SHH) signaling is left unmodulated in the embryonic neocortex of the dorsal forebrain. At early stages of corticogenesis, loss of Suppressor of Fused (Sufu), a potent inhibitor of SHH signaling, in neocortical progenitors, altered their transcriptomic landscape. Ectopic activation of SHH signaling occurred, via degradation of Gli3R, resulting in significant upregulation of Fibroblast Growth Factor 15 (FGF15) gene expression. Consequently, activation of FGF signaling, and its downstream effector the MAPK signaling, facilitated expression of genes characteristic of ventral forebrain progenitors. Our studies identify the importance of modulating extrinsic niche signals such as SHH and FGF15 to maintain the competency and specification program of neocortical progenitors throughout corticogenesis.

SIGNIFICANCE STATEMENTLow levels of FGF15 control progenitor proliferation and differentiation during neocortical development but little is known on how FGF15 expression is maintained. Our studies identified SHH signaling as a critical activator of FGF15 expression during corticogenesis. We found that Sufu, via Gli3R, ensured low levels of FGF15 was expressed to prevent abnormal specification of neocortical progenitors. These studies advance our knowledge on the molecular mechanisms guiding the generation of specific neocortical neuronal lineages, their implications in neurodevelopmental diseases, and may guide future studies on how progenitor cells may be utilized for brain repair.
]]></description>
<dc:creator>Yabut, O. R.</dc:creator>
<dc:creator>Ng, H.-X.</dc:creator>
<dc:creator>Yoon, K.</dc:creator>
<dc:creator>Gomez, H. G.</dc:creator>
<dc:creator>Arela, J. C.</dc:creator>
<dc:creator>Pleasure, S.</dc:creator>
<dc:date>2019-11-14</dc:date>
<dc:identifier>doi:10.1101/841932</dc:identifier>
<dc:title><![CDATA[Combined modulation of SHH and FGF signaling is crucial for maintenance of the neocortical progenitor specification program]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/847582v1?rss=1">
<title>
<![CDATA[
Regional and hemispheric susceptibility of the temporal lobe to FTLD-TDP-43-C pathology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/847582v1?rss=1</link>
<description><![CDATA[
Post-mortem studies show that focal anterior temporal lobe (ATL) neurodegeneration is most often caused by frontotemporal lobar degeneration TDP-43 type C pathology. Clinically, these patients are described with different terms, such as semantic variant primary progressive aphasia (svPPA), semantic dementia (SD), or right temporal variant frontotemporal dementia (FTD) depending on whether the predominant symptoms affect language, semantic knowledge for object or people, or socio-emotional behaviors. ATL atrophy presents with various degrees of lateralization, with right-sided cases considered rarer even though estimation of their prevalence is hampered by the paucity of studies on well-characterized, pathology-proven cohorts. Moreover, it is not clear whether left and right variants show a similar distribution of atrophy within the ATL cross-sectionally and longitudinally.

Here we study the largest cohort to-date of pathology-proven TDP-43-C cases diagnosed during life as svPPA, SD or right temporal variant FTD. We analyzed clinical, cognitive, and neuroimaging data from 30 cases, a subset of which was followed longitudinally. Guided by recent structural and functional parcellation studies, we constructed four bilateral ATL regions of interest (ROIs). The computation of an atrophy lateralization index allowed the comparison of atrophy patterns between the two hemispheres. This led to an automatic, imaging-based classification of the cases as left-predominant or right-predominant. We then compared the two groups in terms of regional atrophy patterns within the ATL ROIs (cross-sectionally) and atrophy progression (longitudinally).

Results showed that 40% of pathology proven cases of TDP-43-C diagnosed with a temporal variant presented with right-lateralized atrophy. Moreover, the findings of our ATL ROI analysis indicated that, irrespective of atrophy lateralization, atrophy distribution within both ATLs follows a medial-to-lateral gradient. Finally, in both left and right cases, atrophy appeared to progress to the contralateral ATL, and from the anterior temporal pole to posterior temporal and orbitofrontal regions.

Taken together, our findings indicate that incipient right predominant ATL atrophy is common in TDP-43-C pathology, and that distribution of damage within the ATLs appears to be the same in left- and right- sided variants. Thus, regardless of differences in clinical phenotype and atrophy lateralization, both temporal variants of FTD should be viewed as a spectrum presentation of the same disease.

Highlights Anterior temporal lobe (ATL) degeneration is most often caused by FTLD-TDP type C pathology

 Cases can present with predominantly left (60%) or right (40%) ATL atrophy

 Within ATLs, medial regions are more vulnerable than lateral ones

 The observed spectrum of clinical phenotypes is driven by atrophy lateralization

 Left and right temporal variants of FTD should be considered the same disease
]]></description>
<dc:creator>Borghesani, V.</dc:creator>
<dc:creator>Battistella, G.</dc:creator>
<dc:creator>Mandelli, M. L.</dc:creator>
<dc:creator>Welch, A.</dc:creator>
<dc:creator>Weis, E.</dc:creator>
<dc:creator>Younes, K.</dc:creator>
<dc:creator>Neuhaus, J.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Miller, Z.</dc:creator>
<dc:creator>Gorno-Tempini, M. L.</dc:creator>
<dc:date>2019-11-20</dc:date>
<dc:identifier>doi:10.1101/847582</dc:identifier>
<dc:title><![CDATA[Regional and hemispheric susceptibility of the temporal lobe to FTLD-TDP-43-C pathology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/847798v1?rss=1">
<title>
<![CDATA[
Taking the sub-lexical route: brain dynamics of reading in the semantic variant of Primary Progressive Aphasia. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/847798v1?rss=1</link>
<description><![CDATA[
Reading aloud requires mapping an orthographic form to a phonological one. The mapping process relies on sub-lexical statistical regularities (e.g., "oo" to |u{square}|) or on learned lexical associations between a specific visual form and a series of sounds (e.g., yacht to /j{square}t/). Computational, neuroimaging, and neuropsychological evidence suggest that sub-lexical, phonological and lexico-semantic processes rely on partially distinct neural substrates: a dorsal (occipito-parietal) and a ventral (occipito-temporal) route, respectively.

Here, we investigated the spatiotemporal features of orthography-to-phonology mapping, capitalizing on the time resolution of magnetoencephalography and the unique clinical model offered by patients with semantic variant of Primary Progressive Aphasia (svPPA). Behaviorally, svPPA patients manifest marked lexico-semantic impairments including difficulties in reading words with exceptional orthographic to phonological correspondence (irregular words). Moreover, they present with focal neurodegeneration in the anterior temporal lobe (ATL), affecting primarily the ventral, occipito-temporal, lexical route. Therefore, this clinical population allows for testing of specific hypotheses on the neural implementation of the dualroute model for reading, such as whether damage to one route can be compensated by over-reliance on the other. To this end, we reconstructed and analyzed time-resolved whole-brain activity in 12 svPPA patients and 12 healthy age-matched controls while reading irregular words (e.g., yacht) and pseudowords (e.g., pook).

Consistent with previous findings that the dorsal route is involved in sub-lexical, phonological processes, in control participants we observed enhanced neural activity over dorsal occipito-parietal cortices for pseudowords, when compared to irregular words. This activation was manifested in the beta-band (12-30 Hz), ramping up slowly over 500 ms after stimulus onset and peaking at [~]800 ms, around response selection and production. Consistent with our prediction, svPPA patients did not exhibit this temporal pattern of neural activity observed in controls this contrast. Furthermore, a direct comparison of neural activity between patients and controls revealed a dorsal spatiotemporal cluster during irregular word reading. These findings suggest that the sub-lexical/phonological route is involved in processing both irregular and pseudowords in svPPA.

Together these results provide further evidence supporting a dual-route model for reading aloud mediated by the interplay between lexico-semantic and sub-lexical/phonological neuro-cognitive systems. When the ventral route is damaged, as in the case of neurodegeneration affecting the ATL, partial compensation appears to be possible by over-recruitment of the slower, serial attention-dependent, dorsal one.

Abbreviated SummaryBorghesani et al. investigate brain dynamics during irregular word reading using magnetoencephalographic imaging in patients with semantic variant of primary progressive aphasia. Due to ventral anterior temporal lobe neurodegeneration, patients show greater reliance of dorsal, occipito-parietal brain regions - providing novel evidence for the interplay between ventral and dorsal routes for reading.
]]></description>
<dc:creator>Borghesani, V.</dc:creator>
<dc:creator>Hinkley, L. B.</dc:creator>
<dc:creator>Ranasinghe, K. G.</dc:creator>
<dc:creator>Thompson, M.</dc:creator>
<dc:creator>Shwe, W.</dc:creator>
<dc:creator>Mizuiri, D.</dc:creator>
<dc:creator>Lauricella, M.</dc:creator>
<dc:creator>Europa, E.</dc:creator>
<dc:creator>Honma, S.</dc:creator>
<dc:creator>Miller, Z.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Vossel, K.</dc:creator>
<dc:creator>Houde, J. F.</dc:creator>
<dc:creator>Gorno-Tempini, M. L.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2019-11-21</dc:date>
<dc:identifier>doi:10.1101/847798</dc:identifier>
<dc:title><![CDATA[Taking the sub-lexical route: brain dynamics of reading in the semantic variant of Primary Progressive Aphasia.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/850909v1?rss=1">
<title>
<![CDATA[
Lgr5+ telocytes are a signaling hub at the intestinal villus tip 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/850909v1?rss=1</link>
<description><![CDATA[
The intestinal epithelium is a structured organ composed of crypts harboring Lgr5+ stem cells, and villi harboring differentiated cells. Spatial transcriptomics have demonstrated profound zonation of epithelial gene expression along the villus axis, but the mechanisms shaping this spatial variability are unknown. Here, we combined laser capture micro-dissection and single cell RNA sequencing to uncover spatially zonated populations of mesenchymal cells along the crypt-villus axis. These included villus tip telocytes (VTTs) that express Lgr5, a gene previously considered a specific crypt epithelial stem cell marker. VTTs are elongated cells that line the villus tip epithelium and signal through Bmp morphogens and the non-canonical Wnt5a ligand. Their ablation strongly perturbs the zonation of enterocyte genes induced at the villus tip. Our study provides a spatially-resolved cell atlas of the small intestinal stroma and exposes Lgr5+ villus tip telocytes as regulators of the epithelial spatial expression programs along the villus axis.
]]></description>
<dc:creator>Bahar Halpern, K.</dc:creator>
<dc:creator>Massalha, H.</dc:creator>
<dc:creator>Zwick, R. K.</dc:creator>
<dc:creator>Moor, A. E.</dc:creator>
<dc:creator>Castillo-Azofeifa, D.</dc:creator>
<dc:creator>Rozenberg, M.</dc:creator>
<dc:creator>Farack, L.</dc:creator>
<dc:creator>Egozi, A.</dc:creator>
<dc:creator>Miller, D. R.</dc:creator>
<dc:creator>Averbukh, I.</dc:creator>
<dc:creator>Harnik, Y.</dc:creator>
<dc:creator>Weinberg-Corem, N.</dc:creator>
<dc:creator>de Sauvage, F. J.</dc:creator>
<dc:creator>Amit, I.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Shoshkes-Carmel, M.</dc:creator>
<dc:creator>Itzkovitz, S.</dc:creator>
<dc:date>2019-11-21</dc:date>
<dc:identifier>doi:10.1101/850909</dc:identifier>
<dc:title><![CDATA[Lgr5+ telocytes are a signaling hub at the intestinal villus tip]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/854265v1?rss=1">
<title>
<![CDATA[
Extended Recognition of the Histone H3 Tail by Histone Demethylase KDM5A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/854265v1?rss=1</link>
<description><![CDATA[
Human lysine demethylase KDM5A is a chromatin modifying enzyme associated with transcriptional regulation due to its ability to catalyze removal of methyl groups from methylated lysine 4 of histone H3 (H3K4me3). Amplification of KDM5A is observed in a number of cancers, including breast cancer, prostate cancer, hepatocellular carcinoma, lung cancer and gastric cancer. In this study, we employed alanine scanning mutagenesis to investigate substrate recognition of KDM5A and identify the H3 tail residues necessary for KDM5A-catalyzed demethylation. Our data show that the H3Q5 residue is critical for substrate recognition by KDM5A. Our data also reveal that the protein-protein interactions between KDM5A and the histone H3 tail extend beyond the amino acids proximal to the substrate mark. Specifically, demethylation activity assays show that deletion or mutation of residues at positions 14-18 on the H3 tail results in an 8-fold increase in the KMapp compared to wild-type 18mer peptide, suggesting this distal epitope is important in histone engagement. Finally, we demonstrate that post-translational modifications on this distal epitope can modulate KDM5A-dependent demethylation. Our findings provide insights into H3K4-specific recognition by KDM5A as well as how chromatin context can regulate KDM5A activity and H3K4 methylation status.
]]></description>
<dc:creator>Petronikolou, N.</dc:creator>
<dc:creator>Longbotham, J. E.</dc:creator>
<dc:creator>Fujimori, D.</dc:creator>
<dc:date>2019-11-25</dc:date>
<dc:identifier>doi:10.1101/854265</dc:identifier>
<dc:title><![CDATA[Extended Recognition of the Histone H3 Tail by Histone Demethylase KDM5A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/860999v1?rss=1">
<title>
<![CDATA[
A minimal CRISPR-Cas3 system for genome engineering 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/860999v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas technologies have provided programmable gene editing tools that have revolutionized research. The leading CRISPR-Cas9 and Cas12a enzymes are ideal for programmed genetic manipulation, however, they are limited for genome-scale interventions. Here, we utilized a Cas3-based system featuring a processive nuclease, expressed endogenously or heterologously, for genome engineering purposes. Using an optimized and minimal CRISPR-Cas3 system (Type I-C) programmed with a single crRNA, large deletions ranging from 7 - 424 kb were generated in Pseudomonas aeruginosa with high efficiency and speed. By comparison, Cas9 yielded small deletions and point mutations. Cas3-generated deletion boundaries were variable in the absence of a homology-directed repair (HDR) template, and successfully and efficiently specified when present. The minimal Cas3 system is also portable; large deletions were induced with high efficiency in Pseudomonas syringae and Escherichia coli using an "all-in-one" vector. Notably, Cas3 generated bi-directional deletions originating from the programmed cut site, which was exploited to iteratively reduce a P. aeruginosa genome by 837 kb (13.5%) using 10 distinct crRNAs. We also demonstrate the utility of endogenous Cas3 systems (Type I-C and I-F) and develop an "anti-anti-CRISPR" strategy to circumvent endogenous CRISPR-Cas inhibitor proteins. CRISPR-Cas3 could facilitate rapid strain manipulation for synthetic biological and metabolic engineering purposes, genome minimization, and the analysis of large regions of unknown function.
]]></description>
<dc:creator>Csorgo, B.</dc:creator>
<dc:creator>Leon, L. M.</dc:creator>
<dc:creator>Chau-Ly, I. J.</dc:creator>
<dc:creator>Vasquez-Rifo, A.</dc:creator>
<dc:creator>Berry, J. D.</dc:creator>
<dc:creator>Mahendra, C.</dc:creator>
<dc:creator>Crawford, E.</dc:creator>
<dc:creator>Lewis, J. D.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2019-12-03</dc:date>
<dc:identifier>doi:10.1101/860999</dc:identifier>
<dc:title><![CDATA[A minimal CRISPR-Cas3 system for genome engineering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.07.506997v1?rss=1">
<title>
<![CDATA[
A discrete mode of endosomal GPCR signaling that does not require β-arrestin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.07.506997v1?rss=1</link>
<description><![CDATA[
Many GPCRs are now recognized to initiate a second phase of G protein (Gs) -dependent signaling through the cAMP cascade after endocytosis. A prevailing current view is that endocytosis-promoted signaling from GPCRs is inherently {beta}-arrestin-dependent because {beta}-arrestin is necessary for receptors to internalize and, for some GPCRs, it also promotes Gs activation on endosomes. Here we revise this view by showing that the vasoactive intestinal peptide receptor 1 (VIPR1), a prototypic secretin-family polypeptide hormone receptor, remains bound to {beta}-arrestin after endocytosis but does not require {beta}-arrestin either to internalize or to generate an endosomal signal. {beta}-arrestin instead resolves the endosomal signal into a temporally separated cAMP peak, and it does so by attenuating signaling from the plasma membrane without detectably affecting the endosomal response. The mechanistic basis for this location-specific difference in {beta}-arrestin function is the formation of distinct VIPR1/{beta}-arrestin complexes at each location. The signal-attenuating complex formed at the plasma membrane does not require GRK-mediated phosphorylation of receptors, while the signaling-neutral complex present on the endosome membrane, in contrast, is GRK-dependent. To our knowledge, the present results provide the first direct demonstration that endosomal GPCR signaling can occur in the complete absence of {beta}-arrestin. They also reveal a discrete role of {beta}-arrestin in sculpting the spatiotemporal profile of cellular GPCR - G protein signaling through the location-specific formation or remodeling of GPCR/{beta}-arrestin complexes.
]]></description>
<dc:creator>Blythe, E. E.</dc:creator>
<dc:creator>Von Zastrow, M.</dc:creator>
<dc:date>2022-09-08</dc:date>
<dc:identifier>doi:10.1101/2022.09.07.506997</dc:identifier>
<dc:title><![CDATA[A discrete mode of endosomal GPCR signaling that does not require β-arrestin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.07.507005v1?rss=1">
<title>
<![CDATA[
Actin-driven protrusions generate rapid long-range membrane tension propagation in cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.07.507005v1?rss=1</link>
<description><![CDATA[
Membrane tension is thought to be a long-range integrator of cell physiology. This role necessitates effective tension transmission across the cell. However, the field remains strongly divided as to whether cell membranes support or resist tension propagation, in part due to a lack of adequate tools for locally manipulating membrane tension. We overcome these limitations by leveraging optogenetics to generate localized actinbased protrusions while concurrently monitoring the propagation of membrane tension using dual-trap optical tweezers. Surprisingly, actin-driven protrusions elicit rapid global membrane tension propagation with little to no attenuation, while forces applied to the cell membrane only do not. We present a simple unifying mechanical model in which mechanical forces that act on both the membrane and actin cortex drive rapid, robust membrane tension propagation.

SummaryMechanical perturbations acting on both actin cortex and plasma membrane drive global membrane tension increase within seconds
]]></description>
<dc:creator>De Belly, H.</dc:creator>
<dc:creator>Yan, S.</dc:creator>
<dc:creator>Borja da Rocha, H.</dc:creator>
<dc:creator>Ichbiah, S.</dc:creator>
<dc:creator>Town, J. P.</dc:creator>
<dc:creator>Turlier, H.</dc:creator>
<dc:creator>Bustamante, C. J.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:date>2022-09-08</dc:date>
<dc:identifier>doi:10.1101/2022.09.07.507005</dc:identifier>
<dc:title><![CDATA[Actin-driven protrusions generate rapid long-range membrane tension propagation in cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.08.507176v1?rss=1">
<title>
<![CDATA[
Potentiation of Adipogenesis by Reactive Oxygen Species is a Unifying Mechanism in the Pro-adipogenic Properties of Bisphenol A and its New Structural Analogues. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.08.507176v1?rss=1</link>
<description><![CDATA[
AimsStructural analogues of bisphenol A (BPA), including BPS and BPF, are emerging environmental toxicants as their presence in the environment is rising since new regulatory restrictions were placed on BPA-containing infant products. The adipogenesis-enhancing effect of bisphenols may explain the link between human exposure and metabolic disease; however, underlying molecular pathways remain unresolved.

ResultsExposure to BPS, BPF, BPA or ROS generators enhanced lipid droplet formation and expression of adipogenic markers after induction of differentiation in adipose-derived progenitors isolated from mice. RNAseq analysis in BPS-exposed progenitors revealed modulation in pathways regulating adipogenesis and responses to oxidative stress. ROS was higher in bisphenol-exposed cells, while co-treatment with antioxidants attenuated adipogenesis and abolished the effect of BPS. There was a loss of mitochondria membrane potential in BPS-exposed cells and mitochondria-derived ROS contributed to potentiation of adipogenesis by BPS and its analogues. Male mice exposed to BPS during gestation had higher whole-body adiposity, as measured by TD-NMR, while postnatal exposure had no impact on adiposity in either sex.

InnovationThese findings support existing evidence showing a role for ROS in regulating adipocyte differentiation and are the first to highlight ROS as a unifying mechanism that explains the pro-adipogenic properties of BPA and its structural analogues.

ConclusionROS act as signaling molecules in the regulation of adipocyte differentiation and mediate bisphenol-induced potentiation of adipogenesis.
]]></description>
<dc:creator>Singh, R. D.</dc:creator>
<dc:creator>Wager, J. L.</dc:creator>
<dc:creator>Scheidl, T. B.</dc:creator>
<dc:creator>Connors, L. T.</dc:creator>
<dc:creator>Easson, S.</dc:creator>
<dc:creator>Callaghan, M. A.</dc:creator>
<dc:creator>Alatorre-hinojosa, S.</dc:creator>
<dc:creator>Swift, L. H.</dc:creator>
<dc:creator>Colarusso, P.</dc:creator>
<dc:creator>Jadli, A.</dc:creator>
<dc:creator>SHUTT, T.</dc:creator>
<dc:creator>Patel, V.</dc:creator>
<dc:creator>Thompson, J.</dc:creator>
<dc:date>2022-09-08</dc:date>
<dc:identifier>doi:10.1101/2022.09.08.507176</dc:identifier>
<dc:title><![CDATA[Potentiation of Adipogenesis by Reactive Oxygen Species is a Unifying Mechanism in the Pro-adipogenic Properties of Bisphenol A and its New Structural Analogues.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.08.507153v1?rss=1">
<title>
<![CDATA[
Mediator 1 ablation induces enamel-to-hair lineage conversion through enhancer dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.08.507153v1?rss=1</link>
<description><![CDATA[
Postnatal cell fate has been postulated to be primarily determined by the local tissue microenvironment. Here, we found that Mediator 1 (Med1) dependent epigenetic mechanisms dictate tissue-specific lineage commitment and progression of dental epithelia. Deletion of Med1, a key component of the Mediator complex linking enhancer activities to gene transcription, provokes a tissue extrinsic lineage shift, causing hair generation in the dental environment. Med1 deficiency gives rise to unusual hair growth via primitive cellular aggregates on incisors. Mechanistically, we found that Med1 establishes super-enhancers that control enamel lineage transcription factors in dental stem cells and their progenies. However, Med1 deficiency reshapes the enhancer landscapes and causes a switch from the dental epithelial transcriptional program towards hair and epidermis on incisors in vivo, and in dental epithelial stem cells in vitro. Med1 loss also provokes an increase in the number and size of enhancers. Interestingly, control dental epithelia already exhibit enhancers for hair and epidermal key transcription factors; these expand in size and transform into active super-enhancers upon Med1 loss suggesting that these epigenetic mechanisms cause the transcriptomic and phenotypic shift towards epidermal and hair lineages. Thus, we propose a role for Med1 in safeguarding lineage specific enhancers, highlight the central role of enhancer accessibility and usage in lineage reprogramming and provide new insights into ectodermal regeneration.
]]></description>
<dc:creator>Thaler, R.</dc:creator>
<dc:creator>Yoshizaki, K.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Fukumoto, S.</dc:creator>
<dc:creator>DenBesten, p.</dc:creator>
<dc:creator>Bikle, D. D.</dc:creator>
<dc:creator>Oda, Y.</dc:creator>
<dc:date>2022-09-09</dc:date>
<dc:identifier>doi:10.1101/2022.09.08.507153</dc:identifier>
<dc:title><![CDATA[Mediator 1 ablation induces enamel-to-hair lineage conversion through enhancer dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.08.506006v1?rss=1">
<title>
<![CDATA[
Beta spectral power during sleep is associated with impaired recall of extinguished fear 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.08.506006v1?rss=1</link>
<description><![CDATA[
The failure to retain memory for extinguished fear plays a major role in the maintenance of post-traumatic stress disorder (PTSD), with successful extinction recall necessary for symptom reduction. Disturbed sleep, a hallmark symptom of PTSD, impairs fear extinction recall. However, our understanding of the electrophysiological mechanisms underpinning sleeps role in extinction retention remain underdetermined. We examined the relationship between the microarchitecture of sleep and extinction recall in healthy humans (n=46, both male and females included) and a pilot study in individuals with PTSD (n=12). Participants underwent a fear conditioning and extinction protocol over two days, with sleep recording occurring between conditioning and extinction. Twenty-four hours after extinction learning, participants underwent extinction recall. Power spectral density (PSD) was computed for pre- and post-extinction learning sleep. Increased beta band PSD ([~]17-26Hz) during pre-extinction learning sleep was associated with worse extinction recall in healthy participants (r=.41, p=.004). Beta PSD was highly stable across three nights of sleep (intraclass correlations (ICC)>0.92). Individuals with PTSD were found to have increased beta PSD compared to healthy controls (ps < .039), and beta PSD correlated with extinction recall in the PTSD group at a similar magnitude to controls (r=.39). Results suggest beta band PSD is elevated in PTSD, and is specifically implicated in difficulties recalling extinguished fear.

Significance statementDisturbed sleep is a hallmark feature of posttraumatic stress disorder (PTSD). Certain neural oscillations that occur during sleep have been shown to be altered in PTSD. These include increased 15-30Hz beta oscillations, which are believed to index cortical hyperarousal. Alongside sleep disturbances, patients also show difficulty in recalling extinguished fear. Although prior studies have linked sleep with extinction retention, no studies have investigated the role that neural oscillations during sleep play in this process. Here, we show in both healthy participants and PTSD patients, that increased beta oscillatory power during sleep is associated with impaired extinction retention. Therefore, this study provides new evidence that electrophysiological changes in the sleep EEG of PTSD patients is also implicated in extinction recall processes.
]]></description>
<dc:creator>Denis, D.</dc:creator>
<dc:creator>Bottary, R.</dc:creator>
<dc:creator>Cunningham, T. J.</dc:creator>
<dc:creator>Drummond, S. P. A.</dc:creator>
<dc:creator>Straus, L. D.</dc:creator>
<dc:date>2022-09-10</dc:date>
<dc:identifier>doi:10.1101/2022.09.08.506006</dc:identifier>
<dc:title><![CDATA[Beta spectral power during sleep is associated with impaired recall of extinguished fear]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.09.507371v1?rss=1">
<title>
<![CDATA[
Heterogeneous migration of neuronal progenitors to the insula shapes the human brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.09.507371v1?rss=1</link>
<description><![CDATA[
The human cerebrum consists of a precise and stereotyped arrangement of lobes, gyri, and connectivity that underlies human cognition. The development of this arrangement is less clear. Current models of radial glial cell migration explain individual gyral formation but fail to explain the global configuration of the cerebral lobes. Moreover, the insula, buried in the depths of the Sylvian fissure, belies conventional models. Here, we show that the insula has unique morphology in adults, that insular morphology and slow volumetric growth emerge during fetal development, and that a novel theory of curved migration is required to explain these findings. We calculated morphologic data in the insula and other lobes in adults (N=107) and in an in utero fetal brain atlas (N=81 healthy fetuses). In utero, the insula grows an order of magnitude slower than the other lobes and demonstrates shallower sulci, less curvature, and less surface complexity both in adults and progressively throughout fetal development. Novel spherical projection analysis demonstrates that the lenticular nuclei obstruct 60-70% of radial pathways from the ventricular zone (VZ) to the insula, forcing a curved migration path to the insula in contrast to a direct radial pathway. Using fetal diffusion tractography, we identify streams of putative progenitor cells that originate from the VZ and migrate tangentially around the lenticular nuclei to form the insula. These results challenge existing models of radial migration to the cortex, provide an alternative model for insular and cerebral development, and lay the groundwork to understand cerebral malformations, insular functional connectivity, and insular pathologies.
]]></description>
<dc:creator>Mallela, A. N.</dc:creator>
<dc:creator>Deng, H.</dc:creator>
<dc:creator>Gholipour, A.</dc:creator>
<dc:creator>Warfield, S. K.</dc:creator>
<dc:creator>Goldschmidt, E.</dc:creator>
<dc:date>2022-09-10</dc:date>
<dc:identifier>doi:10.1101/2022.09.09.507371</dc:identifier>
<dc:title><![CDATA[Heterogeneous migration of neuronal progenitors to the insula shapes the human brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.08.507168v1?rss=1">
<title>
<![CDATA[
Perturbations in podocyte transcriptome and biological pathways induced by FSGS associated circulating factors. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.08.507168v1?rss=1</link>
<description><![CDATA[
Focal segmental glomerulosclerosis (FSGS) is frequently associated with heavy proteinuria and progressive renal failure requiring dialysis or kidney transplantation. However, primary FSGS also has 40-80% risk of recurrence of disease in the transplanted kidney (rFSGS). Multiple circulating factors have been proposed to contribute to the pathogenesis of primary and rFSGS. However, neither the factors nor the downstream effectors specific to individual factors have been identified. The tumor necrosis factor, TNF pathway activation by one or more circulating factors present in the sera of patients with FSGS has been supported by multiple studies. The proposed circulating factors include soluble urokinase-type plasminogen activator receptor (suPAR) and patient derived CD40 autoantibody (CD40autoAb) in the development and recurrence of FSGS. In a human in vitro model, using two novel human antibodies-anti uPAR (2G10) and anti CD40 antibody, we show that the podocyte injury caused by sera from FSGS patients is at least in part mediated by CD40 and suPAR. Additionally, we employ gene expression studies to compare the molecules and pathways activated in response to CD40 autoantibody from rFSGS patients (rFSGS/CD40autoAb) and suPAR, and delineate the unique pathways associated with FSGS injury and transcriptional podocyte alterations with targeted blockade of suPAR and CD40 pathways.

Clinical ImpactFocal Segmental Glomerulosclerosis remains a disease without specific therapy for primary disease and high rate of recurrence after kidney transplantation. Circulating factors are implicated in the pathogenesis of FSGS but targeting them for therapy has remained elusive. We propose two potential therapeutic molecules for rFSGS treatment-a human anti-uPAR antibody (2G10) and a humanized anti-CD40 blocking antibody (Bristol Meyer Squibb, 986090) that reverse podocyte injury associated with FSGS in cultured podocytes and can be further tested in pre-clinical and clinical models. Furthermore, we use microarray profiling to identify transcriptional pathways specific to podocyte injury from patient-derived CD40 autoantibodies (rFSGS/CD40autoAb) and suPAR and selective blockade of these pathways to abrogate podocyte injury.
]]></description>
<dc:creator>Rashmi, P.</dc:creator>
<dc:creator>Sigdel, T. K.</dc:creator>
<dc:creator>Rychkov, D.</dc:creator>
<dc:creator>Damm, I.</dc:creator>
<dc:creator>da Silva, A. A.</dc:creator>
<dc:creator>Vincenti, F.</dc:creator>
<dc:creator>Lourenco, A. L.</dc:creator>
<dc:creator>Craik, C. S.</dc:creator>
<dc:creator>Reiser, J.</dc:creator>
<dc:creator>Sarwal, M. M.</dc:creator>
<dc:date>2022-09-12</dc:date>
<dc:identifier>doi:10.1101/2022.09.08.507168</dc:identifier>
<dc:title><![CDATA[Perturbations in podocyte transcriptome and biological pathways induced by FSGS associated circulating factors.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.10.507433v1?rss=1">
<title>
<![CDATA[
Genetic deletion and pharmacologic inhibition of FcER1 reduces renal injury in mouse models of diabetic nephropathy. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.10.507433v1?rss=1</link>
<description><![CDATA[
FcER1 forms a high affinity multimeric cell-surface receptor for the Fc region of immunoglobulin E (IgE) and controls the activation of mast cells and basophils. Antigen binding and cross-linking of FcER1 associated IgE induces several downstream signaling pathways that result in diverse outcomes. Canonical signaling through IgE-FcER1 has been related to allergic responses, however, recent studies have identified that their function in mast cell and basophils contribute to other pathogenic conditions such as cancer and diabetes. Previous studies have demonstrated that FcER1 protein is upregulated in advanced diabetic kidney disease (DKD) making it a targetable molecule for the treatment of DKD. This study presents evidence that loss of FcER1 signaling reduces proteinuria and renal injury in two pre-clinical mouse models of diabetes. Mice deficient for fcer1 are protected from streptozotocin mediated induction of proteinuria and display reduced fibrosis and mast cell infiltration in kidney. Furthermore, inhibition of FcER1 signaling with an antibody directed against the {gamma}-subunit reduces proteinuria in a spontaneous model of type II diabetes. Our results show significant reduction of proteinuria and tissue damage in pre-clinical DKD models demonstrating the potential of FcER1 inhibitory approaches for developing new therapies in DKD.
]]></description>
<dc:creator>Rashmi, P.</dc:creator>
<dc:creator>Silva, A. A.</dc:creator>
<dc:creator>Sigdel, T.</dc:creator>
<dc:creator>Damm, I.</dc:creator>
<dc:creator>Gouvea, A. L. F.</dc:creator>
<dc:creator>Koliwad, S.</dc:creator>
<dc:creator>Walavalkar, V.</dc:creator>
<dc:creator>Hakroush, S.</dc:creator>
<dc:creator>Sarwal, M. M.</dc:creator>
<dc:date>2022-09-13</dc:date>
<dc:identifier>doi:10.1101/2022.09.10.507433</dc:identifier>
<dc:title><![CDATA[Genetic deletion and pharmacologic inhibition of FcER1 reduces renal injury in mouse models of diabetic nephropathy.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.13.507837v1?rss=1">
<title>
<![CDATA[
Microcosm cultures of a complex synthetic community reveal ecology and genetics of gut microbial organization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.13.507837v1?rss=1</link>
<description><![CDATA[
The behavior of microbial communities depends on both taxonomic composition and physical structure. Metagenomic sequencing of fecal samples has revealed the composition of human gut microbiomes, but we remain less familiar with the spatial organization of microbes between regions such as lumen and mucosa, as well as the microbial genes that regulate this organization. To discover the determinants of spatial organization in the gut, we simulate mucosal colonization over time using an in vitro culture approach incorporating mucin hydrogel microcosms with a complex yet defined community of 123 human strains for which we generated high-quality genome assemblies. Tracking strain abundance longitudinally using shotgun metagenomic measurements, we observe distinct and strain-specific spatial organization in our cultures with strains enriched on mucin microcosms versus in supernatant, reminiscent of mucosa versus lumen enrichment in vivo. Our high taxonomic resolution data enables a comprehensive search for microbial genes that underlie this spatial organization. We identify gene families positively associated with microcosm-enrichment, including several known for biofilm and adhesion functions such as efflux pumps, gene expression regulation, and membrane proteases, as well as a novel link between a coenzyme F420 hydrogenase gene family and lipo/exopolysaccharide biosynthesis. Our strain-resolved abundance measurements also demonstrate that incorporation of microcosms yields a more diverse community than liquid-only culture by allowing co-existence of closely related strains. Altogether these findings demonstrate that microcosm culture with synthetic communities can effectively simulate lumen versus mucosal regions in the gut, providing measurements of microbial organization with high taxonomic resolution to enable identification of specific bacterial genes and functions associated with spatial structure.
]]></description>
<dc:creator>Jin, X.</dc:creator>
<dc:creator>Yu, F. B.</dc:creator>
<dc:creator>Yan, J.</dc:creator>
<dc:creator>Weakley, A.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2022-09-13</dc:date>
<dc:identifier>doi:10.1101/2022.09.13.507837</dc:identifier>
<dc:title><![CDATA[Microcosm cultures of a complex synthetic community reveal ecology and genetics of gut microbial organization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.13.507547v1?rss=1">
<title>
<![CDATA[
Vowel and formant representation in human auditory speech cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.13.507547v1?rss=1</link>
<description><![CDATA[
Vowel sounds are a fundamental component of human speech across all languages. Vowels are cued acoustically by formants, the resonance frequencies determined by the shape of the vocal tract during speaking. An outstanding question in neurolinguistics is how the human brain processes vowel formants during speech perception. We used high-density intracranial recordings from the human speech cortex on the superior temporal gyrus (STG) while participants listened to natural continuous speech to address this question. We derived two-dimensional receptive fields based on the first and second formants to characterize tuning to vowel sounds. We found that neural activity at single STG sites was highly selective for particular zones in the formant space. Furthermore, this formant tuning shifted dynamically to adjust for speaker-specific spectral context. Despite this formant selectivity, local cortical responses were not sufficient to reliably discriminate between vowel categories. Instead, vowel category representations could be accurately decoded when using the entire population of formant encoding electrodes. Together, our results reveal that vowels are locally encoded in STG with complex acoustic tuning in two-dimensional formant space. As a population code this gives rise to phonological vowel perception.
]]></description>
<dc:creator>Oganian, Y.</dc:creator>
<dc:creator>Bhaya-Grossman, I.</dc:creator>
<dc:creator>Johnson, K.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:date>2022-09-14</dc:date>
<dc:identifier>doi:10.1101/2022.09.13.507547</dc:identifier>
<dc:title><![CDATA[Vowel and formant representation in human auditory speech cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.13.507781v1?rss=1">
<title>
<![CDATA[
The landscape of antibody binding affinity in SARS-CoV-2 Omicron BA.1 evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.13.507781v1?rss=1</link>
<description><![CDATA[
The Omicron BA.1 variant of SARS-CoV-2 escapes convalescent sera and monoclonal antibodies that are effective against earlier strains of the virus. This immune evasion is largely a consequence of mutations in the BA.1 receptor binding domain (RBD), the major antigenic target of SARS-CoV-2. Previous studies have identified several key RBD mutations leading to escape from most antibodies. However, little is known about how these escape mutations interact with each other and with other mutations in the RBD. Here, we systematically map these interactions by measuring the binding affinity of all possible combinations of these 15 RBD mutations (215 = 32,768 genotypes) to four monoclonal antibodies (LY-CoV016, LY-CoV555, REGN10987, and S309) with distinct epitopes. We find that BA.1 can lose affinity to diverse antibodies by acquiring a few large-effect mutations and can reduce affinity to others through several small-effect mutations. However, our results also reveal alternative pathways to antibody escape that do not include every large-effect mutation. Moreover, epistatic interactions are shown to constrain affinity decline in S309 but only modestly shape the affinity landscapes of other antibodies. Together with previous work on the ACE2 affinity landscape, our results suggest that escape of each antibody is mediated by distinct groups of mutations, whose deleterious effects on ACE2 affinity are compensated by another distinct group of mutations (most notably Q498R and N501Y).
]]></description>
<dc:creator>Moulana, A.</dc:creator>
<dc:creator>Dupic, T.</dc:creator>
<dc:creator>Phillips, A. M.</dc:creator>
<dc:creator>Chang, J.</dc:creator>
<dc:creator>Roffler, A. A.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Desai, M. M.</dc:creator>
<dc:date>2022-09-14</dc:date>
<dc:identifier>doi:10.1101/2022.09.13.507781</dc:identifier>
<dc:title><![CDATA[The landscape of antibody binding affinity in SARS-CoV-2 Omicron BA.1 evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.14.507933v1?rss=1">
<title>
<![CDATA[
Pbp1 stabilizes and promotes the translation of Puf3-target mRNAs involved in mitochondrial biogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.14.507933v1?rss=1</link>
<description><![CDATA[
Pbp1 (poly(A)-binding protein - binding protein 1) is a cytoplasmic stress granule marker that is capable of forming condensates that function in the negative regulation of TORC1 signaling under respiratory conditions. How mutations in its mammalian ortholog ataxin-2 are linked to neurodegenerative conditions remains unclear. Here, we show that loss of Pbp1 leads to decreases in amounts of mitochondrial proteins whose encoding mRNAs are targets of Puf3, a member of the PUF (Pumilio and FBF) family of RNA-binding proteins. We found that Pbp1 stabilizes and promotes the translation of Puf3-target mRNAs in respiratory conditions, such as those involved in the assembly of cytochrome c oxidase. We further show that Pbp1 and Puf3 interact through their respective low complexity domains, which is required for Puf3-target mRNA stabilization and translation. Our findings reveal a key role for Pbp1-containing assemblies in enabling the translation of mRNAs critical for mitochondrial biogenesis and respiration. They may further begin to explain prior associations of Pbp1/ataxin-2 with RNA, stress granule biology, mitochondrial function, and neuronal health.
]]></description>
<dc:creator>van de Poll, F.</dc:creator>
<dc:creator>Sutter, B. M.</dc:creator>
<dc:creator>Lee, C.-D.</dc:creator>
<dc:creator>Yang, Y.-S.</dc:creator>
<dc:creator>Tu, B. P.</dc:creator>
<dc:date>2022-09-14</dc:date>
<dc:identifier>doi:10.1101/2022.09.14.507933</dc:identifier>
<dc:title><![CDATA[Pbp1 stabilizes and promotes the translation of Puf3-target mRNAs involved in mitochondrial biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.13.507832v1?rss=1">
<title>
<![CDATA[
OpenPBTA: An Open Pediatric Brain Tumor Atlas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.13.507832v1?rss=1</link>
<description><![CDATA[
SummaryPediatric brain and spinal cancer are the leading disease-related cause of death in children, thus we urgently need curative therapeutic strategies for these tumors. To accelerate such discoveries, the Childrens Brain Tumor Network and Pacific Pediatric Neuro-Oncology Consortium created a systematic process for tumor biobanking, model generation, and sequencing with immediate access to harmonized data. We leverage these data to create OpenPBTA, an open collaborative project which establishes over 40 scalable analysis modules to genomically characterize 1,074 pediatric brain tumors. Transcriptomic classification reveals that TP53 loss is a significant marker for poor overall survival in ependymomas and H3 K28-altered diffuse midline gliomas and further identifies universal TP53 dysregulation in mismatch repair-deficient hypermutant high-grade gliomas. OpenPBTA is a foundational analysis platform actively being applied to other pediatric cancers and inform molecular tumor board decision-making, making it an invaluable resource to the pediatric oncology community.

In BriefThe OpenPBTA is a global, collaborative open-science initiative which brought together researchers and clinicians to genomically characterize 1,074 pediatric brain tumors and 22 patient-derived cell lines. Shapiro, et. al create over 40 open-source, scalable modules to perform cancer genomics analyses and provide a richly-annotated somatic dataset across 58 brain tumor histologies. The OpenPBTA framework can be used as a model for large-scale data integration to inform basic research, therapeutic target identification, and clinical translation.

HighlightsOpenPBTA collaborative analyses establish resource for 1,074 pediatric brain tumors NGS-based WHO-aligned integrated diagnoses generated for 641 of 1,074 tumors RNA-Seq analysis infers medulloblastoma subtypes, TP53 status, and telomerase activity OpenPBTA will accelerate therapeutic translation of genomic insights
]]></description>
<dc:creator>Shapiro, J. A.</dc:creator>
<dc:creator>Gaonkar, K. S.</dc:creator>
<dc:creator>Savonen, C. L.</dc:creator>
<dc:creator>Spielman, S. J.</dc:creator>
<dc:creator>Bethell, C. J.</dc:creator>
<dc:creator>Jin, R.</dc:creator>
<dc:creator>Rathi, K. S.</dc:creator>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Egolf, L. E.</dc:creator>
<dc:creator>Farrow, B. K.</dc:creator>
<dc:creator>Miller, D. P.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Koganti, T.</dc:creator>
<dc:creator>Noureen, N.</dc:creator>
<dc:creator>Koptyra, M. P.</dc:creator>
<dc:creator>Duong, N.</dc:creator>
<dc:creator>Kraya, A. A.</dc:creator>
<dc:creator>Kline, C. N.</dc:creator>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Storm, P. B.</dc:creator>
<dc:creator>Nazarian, J.</dc:creator>
<dc:creator>Mack, S. C.</dc:creator>
<dc:creator>Raman, P.</dc:creator>
<dc:creator>Zheng, S.</dc:creator>
<dc:creator>Madsen, P. J.</dc:creator>
<dc:creator>Van Kuren, N.</dc:creator>
<dc:creator>Robins, S.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Waanders, A. J.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Hanson, D.</dc:creator>
<dc:creator>Koschmann, C. J.</dc:creator>
<dc:creator>Diskin, S. J.</dc:creator>
<dc:creator>Lulla, R. R.</dc:creator>
<dc:creator>Brown, M. A.</dc:creator>
<dc:creator>Wong, J.</dc:creator>
<dc:creator>Mason, J. L.</dc:creator>
<dc:creator>Scolaro, L.</dc:creator>
<dc:creator>Kim, M. C.</dc:creator>
<dc:creator>Xie, H. M.</dc:creator>
<dc:creator>Rood, B. R.</dc:creator>
<dc:creator>Cole, K. A.</dc:creator>
<dc:creator>Margol, A. S.</dc:creator>
<dc:creator>Vaksman, Z.</dc:creator>
<dc:creator>Kaufman, R. S</dc:creator>
<dc:date>2022-09-16</dc:date>
<dc:identifier>doi:10.1101/2022.09.13.507832</dc:identifier>
<dc:title><![CDATA[OpenPBTA: An Open Pediatric Brain Tumor Atlas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.16.508223v1?rss=1">
<title>
<![CDATA[
GPCR Signaling Promotes a Stress-induced Genetic Program for Phenoptosis in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.16.508223v1?rss=1</link>
<description><![CDATA[
How an organism dies is a fundamental yet poorly understood question in biology. An organism can die of many causes, including stress-induced phenoptosis, also defined as organismic death that is regulated by its genome-encoded programs. The mechanism of stress-induced phenoptosis is still largely unknown. Here we show that transient but severe freezing-thaw stress (FTS) in C. elegans induces rapid and robust phenoptosis that is regulated by G-protein coupled receptor (GPCR) signaling. RNAi screens identify the GPCR-encoding fshr-1 in mediating transcriptional responses to FTS. FSHR-1 increases ligand interaction upon FTS and activates a cyclic AMP-PKA cascade leading to a genetic program to promote organismic death under severe stress. FSHR-1/GPCR signaling up-regulates the bZIP-type transcription factor ZIP-10, linking FTS to expression of genes involved in lipid remodeling, proteostasis and aging. A mathematical model suggests that genes may promote organismic death under severe stress conditions, potentially benefiting growth of the clonal population with individuals less stressed and more reproductively privileged. Our studies reveal roles of FSHR-1/GPCR-mediated signaling in stress-induced gene expression and phenoptosis in C. elegans, providing empirical new insights into mechanisms of stress-induced phenoptosis with evolutionary implications.
]]></description>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Long, Y.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2022-09-17</dc:date>
<dc:identifier>doi:10.1101/2022.09.16.508223</dc:identifier>
<dc:title><![CDATA[GPCR Signaling Promotes a Stress-induced Genetic Program for Phenoptosis in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.14.506830v1?rss=1">
<title>
<![CDATA[
Quotidian Profile of Vergence Angle in Ambulatory Subjects Monitored with Wearable Eye Tracking Glasses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.14.506830v1?rss=1</link>
<description><![CDATA[
PURPOSEWearable tracking glasses record eye movements and fixations as ambulatory subjects navigate their environment. We tested the performance of eye tracking glasses under laboratory and real world conditions, to characterize the vergence behavior of normal individuals engaged in their customary daily pursuits.

METHODSTo define the accuracy and variability of the eye tracking glasses, 4 subjects fixated with the head stabilized at a series of distances corresponding to vergence demands of: 0.25, 0.50, 1, 2, 4, 8, 16, and 32{degrees}. Then, 10 subjects wore the eye tracking glasses for prolonged periods while carrying out their normal activities. Vergence profiles were compiled for each subject and compared with interpupillary distance.

RESULTSIn the laboratory the eye tracking glasses were comparable in accuracy to remote video eye trackers, outputting a mean vergence value within 1{degrees} of demand at all angles except 32{degrees}. In ambulatory subjects the glasses were less accurate, due to tracking interruptions and measurement errors, only partly mitigated by application of data filters. Nonetheless, a useful record of vergence behavior was obtained in every subject. Vergence angle often had a bimodal distribution, reflecting a preponderance of activities at near (mobile phone, computer) or far (driving, walking). Vergence angle was highly correlated with interpupillary distance.

CONCLUSIONSWearable eye tracking glasses provide a history of vergence angle and the corresponding scene witnessed by ambulatory subjects. They offer insight into the diversity of human ocular motor behavior and may become useful for diagnosis of disorders that affect vergence, such as convergence insufficiency, Parkinson disease, and strabismus.
]]></description>
<dc:creator>Dilbeck, M. D.</dc:creator>
<dc:creator>Gentry, T. N.</dc:creator>
<dc:creator>Economides, J. R.</dc:creator>
<dc:creator>Horton, J. C.</dc:creator>
<dc:date>2022-09-18</dc:date>
<dc:identifier>doi:10.1101/2022.09.14.506830</dc:identifier>
<dc:title><![CDATA[Quotidian Profile of Vergence Angle in Ambulatory Subjects Monitored with Wearable Eye Tracking Glasses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.17.508391v1?rss=1">
<title>
<![CDATA[
A family of novel immune systems targets early infection of nucleus-forming jumbo phages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.17.508391v1?rss=1</link>
<description><![CDATA[
Jumbo bacteriophages of the [ovbar]KZ-like family are characterized by large genomes (>200 kb) and the remarkable ability to assemble a proteinaceous nucleus-like structure. The nucleus protects the phage genome from canonical DNA-targeting immune systems, such as CRISPR-Cas and restriction-modification. We hypothesized that the failure of common bacterial defenses creates selective pressure for immune systems that target the unique jumbo phage biology. Here, we identify the "jumbo phage killer" (Juk) immune system that is deployed by a clinical isolate of Pseudomonas aeruginosa to resist [ovbar]KZ. Juk immunity rescues the cell by preventing early phage transcription, DNA replication, and nucleus assembly. Phage infection is first sensed by JukA (formerly YaaW), which localizes rapidly to the site of phage infection at the cell pole, triggered by ejected phage factors. The effector protein JukB is recruited by JukA, which is required to enable immunity and the subsequent degradation of the phage DNA. JukA homologs are found in several bacterial phyla and are associated with numerous other putative effectors, many of which provided specific anti-[ovbar]KZ activity when expressed in P. aeruginosa. Together, these data reveal a novel strategy for immunity whereby immune factors are recruited to the site of phage protein and DNA ejection to prevent phage progression and save the cell.
]]></description>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Guan, J.</dc:creator>
<dc:creator>Hareendranath, S.</dc:creator>
<dc:creator>Crawford, E.</dc:creator>
<dc:creator>Agard, D. A.</dc:creator>
<dc:creator>Makarova, K. S.</dc:creator>
<dc:creator>Koonin, E. V.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2022-09-18</dc:date>
<dc:identifier>doi:10.1101/2022.09.17.508391</dc:identifier>
<dc:title><![CDATA[A family of novel immune systems targets early infection of nucleus-forming jumbo phages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.16.508329v1?rss=1">
<title>
<![CDATA[
Genetic, clinical underpinnings of subtle early brain change along Alzheimer's dimensions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.16.508329v1?rss=1</link>
<description><![CDATA[
Alzheimers disease (AD) is associated with heterogeneous atrophy patterns. We employed a semi-supervised clustering technique known as Surreal-GAN, through which we identified two dominant dimensions of brain atrophy in symptomatic mild cognitive impairment (MCI) and AD patients: the "diffuse-AD" (R1) dimension shows widespread brain atrophy, and the "MTL-AD" (R2) dimension displays focal medial temporal lobe (MTL) atrophy. Critically, only R2 was associated with widely known sporadic AD genetic risk factors (e.g., APOE {varepsilon}4) in MCI and AD patients at baseline. We then independently detected the presence of the two dimensions in the early stages by deploying the trained model in the general population and two cognitively unimpaired cohorts of asymptomatic participants. In the general population, genome-wide association studies found 77 genes unrelated to APOE differentially associated with R1 and R2. Functional analyses revealed that these genes were overrepresented in differentially expressed gene sets in organs beyond the brain (R1 and R2), including the heart (R1) and the pituitary gland, muscle, and kidney (R2). These genes were enriched in biological pathways implicated in dendritic cells (R2), macrophage functions (R1), and cancer (R1 and R2). Several of them were "druggable genes" for cancer (R1), inflammation (R1), cardiovascular diseases (R1), and diseases of the nervous system (R2). The longitudinal progression showed that APOE {varepsilon}4, amyloid, and tau were associated with R2 at early asymptomatic stages, but this longitudinal association occurs only at late symptomatic stages in R1. Our findings deepen our understanding of the multifaceted pathogenesis of AD beyond the brain. In early asymptomatic stages, the two dimensions are associated with diverse pathological mechanisms, including cardiovascular diseases, inflammation, and hormonal dysfunction - driven by genes different from APOE - which may collectively contribute to the early pathogenesis of AD.
]]></description>
<dc:creator>WEN, J.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Nasrallah, I.</dc:creator>
<dc:creator>Cui, Y.</dc:creator>
<dc:creator>Erus, G.</dc:creator>
<dc:creator>Srinivasan, D.</dc:creator>
<dc:creator>Abdulkadir, A.</dc:creator>
<dc:creator>Mamourian, E.</dc:creator>
<dc:creator>Hwang, G.</dc:creator>
<dc:creator>Singh, A.</dc:creator>
<dc:creator>Bergman, M.</dc:creator>
<dc:creator>Bao, J.</dc:creator>
<dc:creator>Varol, E.</dc:creator>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Boquet-Pujadas, A.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Toga, A. W.</dc:creator>
<dc:creator>Saykin, A. J.</dc:creator>
<dc:creator>Hohman, T.</dc:creator>
<dc:creator>Thompson, P.</dc:creator>
<dc:creator>Villeneuve, S.</dc:creator>
<dc:creator>Gollub, R.</dc:creator>
<dc:creator>Sotiras, A.</dc:creator>
<dc:creator>Wittfeld, K.</dc:creator>
<dc:creator>Grabe, H.</dc:creator>
<dc:creator>Tosun, D.</dc:creator>
<dc:creator>Bilgel, M.</dc:creator>
<dc:creator>An, Y.</dc:creator>
<dc:creator>Marcus, D.</dc:creator>
<dc:creator>LaMontagne, P. J.</dc:creator>
<dc:creator>Heckbert, S.</dc:creator>
<dc:creator>Austin, T.</dc:creator>
<dc:creator>Launer, L.</dc:creator>
<dc:creator>Espeland, M.</dc:creator>
<dc:creator>Masters, C.</dc:creator>
<dc:creator>Maruff, P.</dc:creator>
<dc:creator>Fripp, J.</dc:creator>
<dc:creator>Johnson, S. C.</dc:creator>
<dc:creator>Morris, j.</dc:creator>
<dc:creator>Albert, M.</dc:creator>
<dc:creator>Nick, R.</dc:creator>
<dc:creator>Resnick, S. M.</dc:creator>
<dc:creator>Ferrucci, L.</dc:creator>
<dc:creator>Fan, Y.</dc:creator>
<dc:creator>Mohamad, H.</dc:creator>
<dc:creator>Wolk, D.</dc:creator>
<dc:creator>Shen, L.</dc:creator>
<dc:creator>Shou, H.</dc:creator>
<dc:creator>Dav</dc:creator>
<dc:date>2022-09-19</dc:date>
<dc:identifier>doi:10.1101/2022.09.16.508329</dc:identifier>
<dc:title><![CDATA[Genetic, clinical underpinnings of subtle early brain change along Alzheimer's dimensions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.19.508328v1?rss=1">
<title>
<![CDATA[
Homeostatic cytokines reciprocally modulate the emergence of prenatal effector PLZF+CD4+ T cells in humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.19.508328v1?rss=1</link>
<description><![CDATA[
The development of human adaptive immunity progresses faster than previously appreciated, with the emergence of memory CD4+ T cells alongside regulatory T (Treg) cells by the second trimester of pregnancy. We previously identified a prenatal-specific subset of PLZF+ CD4+ T cells with heightened effector potential that accounted for most memory T cells in the developing intestine and accumulated in the cord blood of infants exposed to prenatal inflammatory pathologies. However, the signals that drive their tissue distribution and effector maturation are unknown. In this report, we define the transcriptional and functional heterogeneity of prenatal PLZF+ CD4+ T cells and identify compartmentalization of Th-like effector function across the small intestine (SI) and mesenteric lymph nodes (MLN). We find that IL-7, which is more abundant in the SI relative to the MLN, drives the preferential expansion of naive PLZF+ CD4+ T cells via JAK/STAT and MEK/ERK signaling. Exposure to IL-7 induces a subset of PLZF+ CD4+ T cells to acquire a memory-phenotype and rapid effector function, identifying the human analogue of memory-phenotype CD4+ T cells. Further, IL-7 modulates the differentiation of Th1- and Th17-like PLZF+ CD4+ T cells, and thus likely contributes to the anatomic compartmentalization of prenatal CD4+ T cell effector function.
]]></description>
<dc:creator>Locher, V.</dc:creator>
<dc:creator>Park, S.</dc:creator>
<dc:creator>Bunis, D.</dc:creator>
<dc:creator>Makredes, S.</dc:creator>
<dc:creator>Burt, T. D.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Halkias, J.</dc:creator>
<dc:date>2022-09-19</dc:date>
<dc:identifier>doi:10.1101/2022.09.19.508328</dc:identifier>
<dc:title><![CDATA[Homeostatic cytokines reciprocally modulate the emergence of prenatal effector PLZF+CD4+ T cells in humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.19.508335v1?rss=1">
<title>
<![CDATA[
Systems biology illuminates alternative metabolic niches in the human gut microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.19.508335v1?rss=1</link>
<description><![CDATA[
Human gut bacteria perform diverse metabolic functions with consequences for host health. The prevalent and disease-linked Actinobacterium Eggerthella lenta performs several unusual chemical transformations, but it does not metabolize sugars and its core growth strategy remains unclear. To obtain a comprehensive view of the metabolic network of E. lenta, we generated several complementary resources: defined culture media, metabolomics profiles of strain isolates, and a curated genome-scale metabolic reconstruction. Stable isotope-resolved metabolomics revealed that E. lenta uses acetate as a key carbon source while catabolizing arginine to generate ATP, traits which could be recapitulated in silico by our updated metabolic model. We compared these in vitro findings with metabolite shifts observed in E. lenta-colonized gnotobiotic mice, identifying shared signatures across environments and highlighting catabolism of the host signaling metabolite agmatine as an alternative energy pathway. Together, our results elucidate a distinctive metabolic niche filled by E. lenta in the gut ecosystem.
]]></description>
<dc:creator>Noecker, C.</dc:creator>
<dc:creator>Sanchez, J.</dc:creator>
<dc:creator>Bisanz, J. E.</dc:creator>
<dc:creator>Escalante, V.</dc:creator>
<dc:creator>Alexander, M.</dc:creator>
<dc:creator>Trepka, K.</dc:creator>
<dc:creator>Heinken, A.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Dodd, D.</dc:creator>
<dc:creator>Thiele, I.</dc:creator>
<dc:creator>DeFelice, B.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:date>2022-09-19</dc:date>
<dc:identifier>doi:10.1101/2022.09.19.508335</dc:identifier>
<dc:title><![CDATA[Systems biology illuminates alternative metabolic niches in the human gut microbiome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.21.508900v1?rss=1">
<title>
<![CDATA[
Hierarchical sequence-affinity landscapes shape the evolution of breadth in an anti-influenza receptor binding site antibody 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.21.508900v1?rss=1</link>
<description><![CDATA[
Broadly neutralizing antibodies (bnAbs) that neutralize diverse variants of a particular virus are of considerable therapeutic interest1. Recent advances have enabled us to isolate and engineer these antibodies as therapeutics, but eliciting them through vaccination remains challenging, in part due to our limited understanding of how antibodies evolve breadth2. Here, we analyze the landscape by which an anti-influenza receptor binding site (RBS) bnAb, CH65, evolved broad affinity to diverse H1 influenza strains3, 4. We do this by generating an antibody library of all possible evolutionary intermediates between the unmutated common ancestor (UCA) and the affinity-matured CH65 antibody and measure the affinity of each intermediate to three distinct H1 antigens. We find that affinity to each antigen requires a specific set of mutations - distributed across the variable light and heavy chains - that interact non-additively (i.e., epistatically). These sets of mutations form a hierarchical pattern across the antigens, with increasingly divergent antigens requiring additional epistatic mutations beyond those required to bind less divergent antigens. We investigate the underlying biochemical and structural basis for these hierarchical sets of epistatic mutations and find that epistasis between heavy chain mutations and a mutation in the light chain at the VH-VL interface is essential for binding a divergent H1. Collectively, this work is the first to comprehensively characterize epistasis between heavy and light chain mutations and shows that such interactions are both strong and widespread. Together with our previous study analyzing a different class of anti-influenza antibodies5, our results implicate epistasis as a general feature of antibody sequence-affinity landscapes that can potentiate and constrain the evolution of breadth.
]]></description>
<dc:creator>Phillips, A. M.</dc:creator>
<dc:creator>Maurer, D. P.</dc:creator>
<dc:creator>Brooks, C.</dc:creator>
<dc:creator>Dupic, T.</dc:creator>
<dc:creator>Schmidt, A. G.</dc:creator>
<dc:creator>Desai, M. M.</dc:creator>
<dc:date>2022-09-21</dc:date>
<dc:identifier>doi:10.1101/2022.09.21.508900</dc:identifier>
<dc:title><![CDATA[Hierarchical sequence-affinity landscapes shape the evolution of breadth in an anti-influenza receptor binding site antibody]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.20.508732v1?rss=1">
<title>
<![CDATA[
T cell-instructed inflammation drives immune checkpoint inhibitor therapy resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.20.508732v1?rss=1</link>
<description><![CDATA[
Resistance to immune checkpoint inhibitors (ICIs) is common, even in tumors with T cell infiltration. We thus investigated consequences of ICI-induced T cell infiltration in the microenvironment of resistant tumors. T cells increased in ICI-resistant tumors following treatment as did neutrophils, in contrast to ICI-responsive tumors. Resistant tumors were distinguished by high expression of IL-1 Receptor 1 (IL1R1), enabling a synergistic response to IL-1 and TNF to induce G-CSF, CXCL1, and CXCL2 via NF-{kappa}B signaling, supporting neutrophils. Perturbation of this inflammatory resistance circuit sensitized tumors to ICIs. Paradoxically, T cells drove this resistance circuit via TNF both in vitro and in vivo. Evidence of this inflammatory resistance circuit and its impact also translated to human cancers. These data support a novel mechanism of ICI resistance, wherein treatment-induced T cell activity can drive resistance in tumors responsive to IL-1 and TNF, with important therapeutic implications.

Statement of SignificanceAlthough T cell-infiltrated cancers are frequently resistant to immune checkpoint inhibitor therapies, mechanisms of resistance beyond T cell exhaustion remain unclear. Here, we reveal the functional significance of tumor- infiltrating T cells in resistant tumors, which surprisingly instruct immunosuppressive inflammation in mouse and human cancers responsive to IL-1 and TNF.
]]></description>
<dc:creator>Cho, N. W.</dc:creator>
<dc:creator>Guldberg, S. M.</dc:creator>
<dc:creator>Kim, E. J.</dc:creator>
<dc:creator>Hiam-Galvez, K. J.</dc:creator>
<dc:creator>Wai, K.</dc:creator>
<dc:creator>Levine, L. S.</dc:creator>
<dc:creator>Nabet, B. Y.</dc:creator>
<dc:creator>DeBarge, R.</dc:creator>
<dc:creator>Yee, J. L.</dc:creator>
<dc:creator>Ashitey, N. A.</dc:creator>
<dc:creator>Tenvooren, I.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:date>2022-09-22</dc:date>
<dc:identifier>doi:10.1101/2022.09.20.508732</dc:identifier>
<dc:title><![CDATA[T cell-instructed inflammation drives immune checkpoint inhibitor therapy resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.21.508943v1?rss=1">
<title>
<![CDATA[
The Phosphate Deprivation Response is Mediated by an Interaction between Brassinosteroid Signaling and Zinc in Tomato 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.21.508943v1?rss=1</link>
<description><![CDATA[
Phosphate is a necessary macronutrient for basic biological processes, plant growth, and agriculture. Plants modulate their root system architecture and cellular processes to adapt to phosphate deprivation albeit with a growth penalty. Excess application of phosphate fertilizer, on the other hand, leads to eutrophication and has a negative environmental impact. Moreover, phosphate mined from rock reserves is a finite and non-recyclable resource and its levels are nearing complete depletion. Here, we show that Solanum pennellii, a wild relative of tomato, is partially insensitive to phosphate deprivation. Furthermore, it mounts a constitutive response under phosphate sufficiency. We demonstrate that activated brassinosteroid signaling through a tomato BZR1 ortholog gives rise to the same constitutive phosphate deficiency response, which is dependent on zinc over-accumulation. Collectively, these results reveal an additional strategy by which plants can adapt to phosphate starvation.
]]></description>
<dc:creator>Demirer, G. S.</dc:creator>
<dc:creator>Gibson, D. J.</dc:creator>
<dc:creator>Yue, X.</dc:creator>
<dc:creator>Pan, K.</dc:creator>
<dc:creator>Elishav, E.</dc:creator>
<dc:creator>Khandal, H.</dc:creator>
<dc:creator>Horev, G.</dc:creator>
<dc:creator>Tarkowska, D.</dc:creator>
<dc:creator>Canto-Pastor, A.</dc:creator>
<dc:creator>Kong, S.</dc:creator>
<dc:creator>Maloof, J.</dc:creator>
<dc:creator>Savaldi-Goldstein, S.</dc:creator>
<dc:creator>Brady, S.</dc:creator>
<dc:date>2022-09-22</dc:date>
<dc:identifier>doi:10.1101/2022.09.21.508943</dc:identifier>
<dc:title><![CDATA[The Phosphate Deprivation Response is Mediated by an Interaction between Brassinosteroid Signaling and Zinc in Tomato]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.22.508962v1?rss=1">
<title>
<![CDATA[
Pharmacological inhibition of bromodomain and extra-terminal proteins induces NRF-2-mediated inhibition of SARS-CoV-2 replication and is subject to viral antagonism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.22.508962v1?rss=1</link>
<description><![CDATA[
Inhibitors of bromodomain and extra-terminal proteins (iBETs), including JQ-1, have been suggested as potential therapeutics against SARS-CoV-2 infection. However, molecular mechanisms underlying JQ-1-induced antiviral activity and its susceptibility to viral antagonism remain incompletely understood. iBET treatment transiently inhibited infection by SARS-CoV-2 variants and SARS-CoV, but not MERS-CoV. Our functional assays confirmed JQ-1-mediated downregulation of ACE2 expression and multi-omics analysis uncovered induction of an antiviral NRF-2-mediated cytoprotective response as an additional antiviral component of JQ-1 treatment. Serial passaging of SARS-CoV-2 in the presence of JQ-1 resulted in predominance of ORF6-deficient variants. JQ-1 antiviral activity was transient in human bronchial airway epithelial cells (hBAECs) treated prior to infection and absent when administered therapeutically. We propose that JQ-1 exerts pleiotropic effects that collectively induce a transient antiviral state that is ultimately nullified by an established SARS-CoV-2 infection, raising questions on their clinical suitability in the context of COVID-19.
]]></description>
<dc:creator>Mhlekude, B.</dc:creator>
<dc:creator>Postmus, D.</dc:creator>
<dc:creator>Weiner, J.</dc:creator>
<dc:creator>Stenzel, S.</dc:creator>
<dc:creator>Zapatero-Belinchon, F. J.</dc:creator>
<dc:creator>Olmer, R.</dc:creator>
<dc:creator>Jansen, J.</dc:creator>
<dc:creator>Richter, A.</dc:creator>
<dc:creator>Heinze, J.</dc:creator>
<dc:creator>Heinemann, N.</dc:creator>
<dc:creator>Muhlemann, B.</dc:creator>
<dc:creator>Schroeder, S.</dc:creator>
<dc:creator>Jones, T. C.</dc:creator>
<dc:creator>Muller, M. A.</dc:creator>
<dc:creator>Drosten, C.</dc:creator>
<dc:creator>Pich, A.</dc:creator>
<dc:creator>Thiel, V.</dc:creator>
<dc:creator>Martin, U.</dc:creator>
<dc:creator>Niemeyer, D.</dc:creator>
<dc:creator>Gerold, G.</dc:creator>
<dc:creator>Beule, D.</dc:creator>
<dc:creator>Goffinet, C.</dc:creator>
<dc:date>2022-09-23</dc:date>
<dc:identifier>doi:10.1101/2022.09.22.508962</dc:identifier>
<dc:title><![CDATA[Pharmacological inhibition of bromodomain and extra-terminal proteins induces NRF-2-mediated inhibition of SARS-CoV-2 replication and is subject to viral antagonism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.22.509085v1?rss=1">
<title>
<![CDATA[
Growth cone advance requires EB1 as revealed by genomic replacement with a light-sensitive variant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.22.509085v1?rss=1</link>
<description><![CDATA[
A challenge in analyzing dynamic intracellular cell biological processes is the dearth of methodologies that are sufficiently fast and specific to perturb intracellular protein activities. We previously developed a light-sensitive variant of the microtubule plus end tracking protein EB1 by inserting a blue light-controlled protein dimerization module between functional domains. Here we describe an advanced method to replace endogenous EB1 with this light-sensitive variant in a single genome editing step enabling this approach in human induced pluripotent stem cells and derived neurons. We demonstrate that acute and local optogenetic EB1 inactivation in developing cortical neurons induces microtubule depolymerization in the growth cone periphery and subsequent neurite retraction. In addition, advancing growth cones are repelled from areas of blue light exposure. These phenotypes were independent of the neuronal EB1 homolog EB3, revealing a direct dynamic role of EB1-mediated microtubule plus end interactions in neuron morphogenesis and neurite guidance.
]]></description>
<dc:creator>Dema, A.</dc:creator>
<dc:creator>Charafeddine, R. A.</dc:creator>
<dc:creator>Rahgozar, S.</dc:creator>
<dc:creator>van Haren, J.</dc:creator>
<dc:creator>Wittmann, T.</dc:creator>
<dc:date>2022-09-23</dc:date>
<dc:identifier>doi:10.1101/2022.09.22.509085</dc:identifier>
<dc:title><![CDATA[Growth cone advance requires EB1 as revealed by genomic replacement with a light-sensitive variant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.23.509287v1?rss=1">
<title>
<![CDATA[
The ancestral ESCRT protein TOM1L2 selects ubiquitinated cargoes for retrieval from cilia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.23.509287v1?rss=1</link>
<description><![CDATA[
Many G protein-coupled receptors (GPCRs) reside within cilia of mammalian cells and must undergo regulated exit from cilia for the appropriate transduction of signals such as Hedgehog morphogens. Lysine 63-linked ubiquitin (K63Ub) chains mark GPCRs for regulated removal from cilia, but the molecular basis of K63Ub recognition inside cilia remains elusive. Here we show that the BBSome -the trafficking complex in charge of retrieving GPCRs from cilia- engages the ancestral endosomal sorting factor TOM1L2 (Target of Myb1-Like 2) to recognize UbK63 chains within cilia. TOM1L2 directly binds to UbK63 chains and to the BBSome and targeted disruption of the TOM1L2/BBSome interaction results in the accumulation of TOM1L2, ubiquitin and the GPCRs SSTR3, Smoothened and GPR161 inside cilia. Strikingly, the single cell alga Chlamydomonas also requires its TOM1L2 orthologue to clear ubiquitinated proteins from cilia. We conclude that TOM1L2 broadly enables the retrieval of UbK63-tagged proteins by the ciliary trafficking machinery.
]]></description>
<dc:creator>Shinde, S. R.</dc:creator>
<dc:creator>Mick, D. U.</dc:creator>
<dc:creator>Aoki, E.</dc:creator>
<dc:creator>Nachury, M. V.</dc:creator>
<dc:date>2022-09-25</dc:date>
<dc:identifier>doi:10.1101/2022.09.23.509287</dc:identifier>
<dc:title><![CDATA[The ancestral ESCRT protein TOM1L2 selects ubiquitinated cargoes for retrieval from cilia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.24.509337v1?rss=1">
<title>
<![CDATA[
Small-molecule ketone esters treat brain network abnormalities in an Alzheimer's disease mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.24.509337v1?rss=1</link>
<description><![CDATA[
Altered brain network activity and the resulting hypersynchrony are important for the pathogenesis of cognitive decline in Alzheimers disease (AD) mouse models. Treatments that reduce epileptiform discharges (EDs) or network hyperactivity improve cognition in AD models and humans. We first show that ketogenic diet, but not fasting, rapidly and persistently reduced EDs in the hAPPJ20 Alzheimers mouse model over timescales of hours to months. Then, to identify the specific mechanism of the pleiotropic ketogenic diet, we developed small molecule ketone esters to deliver ketone bodies pharmacologically. Two ketone esters recapitulate ED suppression without other dietary manipulation, over time scales of minutes to one week. This small molecule rescue was associated with reduced low-frequency oscillatory activity similar to the recently reported mechanism of an NMDA receptor modulator molecule in this model. Long-term KD resulted in cognitive improvement and in a sex-stratified analysis also improved survival in the more severely affected hAPPJ20 males. Agents that deliver ketone bodies via small molecules or act on downstream targets may hold therapeutic promise in AD through the mechanism of improved network function and reduced epileptiform activity.
]]></description>
<dc:creator>Newman, J. C.</dc:creator>
<dc:creator>Ma, K.</dc:creator>
<dc:creator>Kroll, F.</dc:creator>
<dc:creator>Higgins, E.</dc:creator>
<dc:creator>Ulrich, S.</dc:creator>
<dc:creator>Palop, J. J.</dc:creator>
<dc:creator>Verdin, E.</dc:creator>
<dc:date>2022-09-26</dc:date>
<dc:identifier>doi:10.1101/2022.09.24.509337</dc:identifier>
<dc:title><![CDATA[Small-molecule ketone esters treat brain network abnormalities in an Alzheimer's disease mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.25.509440v1?rss=1">
<title>
<![CDATA[
A non-ligand surrogate agonist antibody that enhances canonical Wnt signaling and bone regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.25.509440v1?rss=1</link>
<description><![CDATA[
The Wnt signaling pathway promotes tissue regeneration and is a promising therapeutic target for treatment of osteolytic bone diseases. Here we report the discovery of a novel type of canonical Wnt agonist antibody that does not operate as a ligand surrogate. The antibody increases Wnt/{beta}-catenin signaling with or without exogenously provided Wnt ligands. It binds to a site on the P3 domain of LRP6 that is distinct from where the Wnt3a ligand and the DKK1 antagonist bind. The agonist effect persists in the presence of DKK1 and is further amplified by R-spondin even when Wnt ligands are not provided, suggesting a potential use for this antibody in ligand-low or insufficient settings. The antibody induces osteoblastic differentiation and mineralization in vitro and restores bone loss in vivo in a myeloma-derived intrafemoral mouse model, opening a potential path for therapeutic development in osteolytic diseases caused by cancer and aging.
]]></description>
<dc:creator>Lee, N.-K.</dc:creator>
<dc:creator>Bidlingmaier, S.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:date>2022-09-27</dc:date>
<dc:identifier>doi:10.1101/2022.09.25.509440</dc:identifier>
<dc:title><![CDATA[A non-ligand surrogate agonist antibody that enhances canonical Wnt signaling and bone regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.23.509256v1?rss=1">
<title>
<![CDATA[
Cell circuits between leukemic cells and mesenchymal stem cells block lymphopoiesis by activating lymphotoxin-beta receptor signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.23.509256v1?rss=1</link>
<description><![CDATA[
Acute lymphoblastic and myeloblastic leukemias (ALL and AML) have been known to modify the bone marrow microenvironment and disrupt non-malignant hematopoiesis. However, the molecular mechanisms driving these alterations remain poorly defined. Here we show that leukemic cells turn-off lymphopoiesis and erythropoiesis shortly after colonizing the bone marrow. ALL and AML cells express lymphotoxin-1{beta}2 and activate LT{beta}R signaling in mesenchymal stem cells (MSCs), which turns off IL7 production and prevents non-malignant lymphopoiesis. We show that the DNA damage response pathway and CXCR4 signaling promote lymphotoxin-1{beta}2 expression in leukemic cells. Genetic or pharmacologic disruption of LT{beta}R signaling in MSCs restores lymphopoiesis but not erythropoiesis, reduces leukemic cell growth, and significantly extends the survival of transplant recipients. Similarly, CXCR4 blocking also prevents leukemia-induced IL7 downregulation, and inhibits leukemia growth. These studies demonstrate that acute leukemias exploit physiological mechanisms governing hematopoietic output as a strategy for gaining competitive advantage.

One Sentence SummaryLeukemias colonize bone marrow niches and disrupt hematopoiesis. However, the cross-talk between leukemia and niche cells remains poorly understood. We show that leukemia activates LT{beta}R in mesenchymal stem cells which suppresses IL7 production and IL7-dependent lymphopoiesis and accelerates leukemia growth.
]]></description>
<dc:creator>Feng, X.</dc:creator>
<dc:creator>Sun, R.</dc:creator>
<dc:creator>Lee, M.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:creator>Geng, H.</dc:creator>
<dc:creator>Müschen, M.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Pereira, J. P.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.23.509256</dc:identifier>
<dc:title><![CDATA[Cell circuits between leukemic cells and mesenchymal stem cells block lymphopoiesis by activating lymphotoxin-beta receptor signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.27.509541v1?rss=1">
<title>
<![CDATA[
A Genome Scale Transcriptional Regulatory Model of the Human Placenta 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.27.509541v1?rss=1</link>
<description><![CDATA[
Gene regulation is essential to placental function and fetal development. We report a genome-scale transcriptional regulatory network (TRN) of the human placenta built using digital genomic footprinting and transcriptomic data. We integrated 475 transcriptomes and 12 DNase hypersensitivity datasets from placental samples to globally and quantitatively map transcription factor (TF)-target gene interactions. In an independent dataset, the TRN model predicted target gene expression with an out of sample R2 value greater than 0.25 for 74% of target genes. We performed siRNA knockdowns of 4 TFs and achieved concordance between the predicted gene targets in our TRN and differences in expression of knockdowns with an accuracy of >0.7 for 3 of the 4 TFs. Our final model contained 113,158 interactions across 391 TFs and 7,712 target genes and is publicly available. We identified six TFs which were significantly enriched as regulators for genes previously associated with preterm birth.
]]></description>
<dc:creator>Paquette, A. G.</dc:creator>
<dc:creator>Ahuna, K.</dc:creator>
<dc:creator>Hwang, Y. M.</dc:creator>
<dc:creator>Pearl, J.</dc:creator>
<dc:creator>Liao, H.</dc:creator>
<dc:creator>Shannon, P.</dc:creator>
<dc:creator>Kadam, L.</dc:creator>
<dc:creator>Lapehn, S.</dc:creator>
<dc:creator>Bucher, M.</dc:creator>
<dc:creator>Roper, R.</dc:creator>
<dc:creator>Funk, C. C.</dc:creator>
<dc:creator>MacDonald, J.</dc:creator>
<dc:creator>Bammler, T.</dc:creator>
<dc:creator>Baloni, P.</dc:creator>
<dc:creator>Brockway, H.</dc:creator>
<dc:creator>Mason, W. A.</dc:creator>
<dc:creator>Bush, N.</dc:creator>
<dc:creator>LeWinn, K. Z.</dc:creator>
<dc:creator>Carr, C. J.</dc:creator>
<dc:creator>Stamatoyannopoulos, J.</dc:creator>
<dc:creator>Muglia, L. J.</dc:creator>
<dc:creator>Jones, H. M.</dc:creator>
<dc:creator>Sadovsky, Y.</dc:creator>
<dc:creator>Myatt, L.</dc:creator>
<dc:creator>Sathyanarayana, S.</dc:creator>
<dc:creator>Price, N.</dc:creator>
<dc:creator>Environmental influences on Child Health Outcomes,</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509541</dc:identifier>
<dc:title><![CDATA[A Genome Scale Transcriptional Regulatory Model of the Human Placenta]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.27.509770v1?rss=1">
<title>
<![CDATA[
A scalable screening platform for phenotypic subtyping of ALS patient-derived fibroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.27.509770v1?rss=1</link>
<description><![CDATA[
A major challenge for understanding and treating Amyotrophic Lateral Sclerosis (ALS) is that most patients have no known genetic cause. Even within defined genetic subtypes, patients display considerable clinical heterogeneity. It is unclear how to identify subsets of ALS patients that share common molecular dysregulation or could respond similarly to treatment. Here, we developed a scalable microscopy and machine learning platform to phenotypically subtype readily available, primary patient-derived fibroblasts. Application of our platform identified robust signatures for the genetic subtype FUS-ALS, allowing cell lines to be scored along a spectrum from FUS-ALS to non-ALS. Our FUS-ALS phenotypic score negatively correlates with age of diagnosis and provides information that is distinct from transcript profiling. Interestingly, the FUS-ALS phenotypic score can be used to identify sporadic patient fibroblasts that have consistent pathway dysregulation with FUS-ALS. Further, we showcase how the score can be used to evaluate the effects of ASO treatment on patient fibroblasts. Our platform provides an approach to move from genetic to phenotypic subtyping and a first step towards rational selection of patient subpopulations for targeted therapies.
]]></description>
<dc:creator>Kumbier, K.</dc:creator>
<dc:creator>Roth, M.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Lazzari-Dean, J.</dc:creator>
<dc:creator>Waters, C.</dc:creator>
<dc:creator>Huang, P.</dc:creator>
<dc:creator>Korobeynikov, V.</dc:creator>
<dc:creator>Consortium, N. Y. G. C.</dc:creator>
<dc:creator>Phatnani, H.</dc:creator>
<dc:creator>Schneider, N.</dc:creator>
<dc:creator>Jacobson, M.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Altschuler, S.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509770</dc:identifier>
<dc:title><![CDATA[A scalable screening platform for phenotypic subtyping of ALS patient-derived fibroblasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.28.509838v1?rss=1">
<title>
<![CDATA[
Genetic Regulators of Sputum Mucin Concentration and Their Associations with COPD Phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.28.509838v1?rss=1</link>
<description><![CDATA[
Hyper-secretion and/or hyper-concentration of mucus is a defining feature of multiple obstructive lung diseases, including chronic obstructive pulmonary disease (COPD). Mucus itself is composed of a mixture of water, ions, salt and proteins, of which the gel-forming mucins, MUC5AC and MUC5B, are the most abundant. Recent studies have linked the concentrations of these proteins in sputum to COPD phenotypes, including chronic bronchitis (CB) and acute exacerbations (AE). We sought to determine whether common genetic variants influence sputum mucin concentrations and whether these variants are also associated with COPD phenotypes, specifically CB and AE. We performed a GWAS to identify quantitative trait loci for sputum mucin protein concentration (pQTL) in the Sub-Populations and InteRmediate Outcome Measures in COPD Study (SPIROMICS, n=708 for total mucin, n=215 for MUC5AC, MUC5B). Subsequently, we tested for associations of mucin pQTL with CB and AE using regression modeling (n=822-1300). Replication analysis was conducted using data from COPDGene (n =5740) and by examining results from the UK Biobank. We identified one genome-wide significant pQTL for MUC5AC (rs75401036) and two for MUC5B (rs140324259, rs10001928). The strongest association for MUC5B, with rs140324259 on chromosome 11, explained 14% of variation in sputum MUC5B. Despite being associated with lower MUC5B, the C allele of rs140324259 conferred increased risk of CB (odds ratio (OR) = 1.42; 95% confidence interval (CI): 1.10-1.80) as well as AE ascertained over three years of follow up (OR=1.41; 95% CI: 1.02-1.94). Associations between rs140324259 and CB or AE did not replicate in COPDGene. However, in the UK Biobank, rs140324259 was associated with phenotypes that define CB, namely chronic mucus production and cough, again with the C allele conferring increased risk. We conclude that sputum MUC5AC and MUC5B concentrations are associated with common genetic variants, and the top locus for MUC5B may influence COPD phenotypes, in particular CB.

Author SummaryChronic obstructive pulmonary disease (COPD) is characterized by presence of emphysema and/or chronic bronchitis. Excessive mucus production is a defining phenotype of chronic bronchitis, and is associated with several important features of COPD, including exacerbations and loss of lung function. Recent studies have demonstrated that the amount of mucus produced in COPD patients is an important marker of disease state. We investigated whether common genetic variants are associated with the concentration of two key proteins in mucus, MUC5AC and MUC5B, and whether the variants we identified are also associated with COPD outcomes. We identified multiple genetic variants that were associated with MUC5AC or MUC5B concentration. The strongest association we detected, for MUC5B on chromosome 11, was also associated with features of COPD, including chronic bronchitis and acute exacerbations, in one COPD study population but not another. Results from a much larger study, the UK Biobank, indicate that this variant is associated with chronic mucus production and chronic cough, which are key features of chronic bronchitis. Thus, we conclude that the concentration of key proteins in mucus are influenced by genetic variation, and that a variant on chromosome 11 that affects MUC5B may in turn alter COPD outcomes.
]]></description>
<dc:creator>Van Buren, E.</dc:creator>
<dc:creator>Radicioni, G.</dc:creator>
<dc:creator>Lester, S.</dc:creator>
<dc:creator>O'Neal, W.</dc:creator>
<dc:creator>Dang, H.</dc:creator>
<dc:creator>Kasela, S.</dc:creator>
<dc:creator>Gardadri, S.</dc:creator>
<dc:creator>Curtis, J.</dc:creator>
<dc:creator>Han, M.</dc:creator>
<dc:creator>Krishnan, J.</dc:creator>
<dc:creator>Wan, E.</dc:creator>
<dc:creator>Silverman, E.</dc:creator>
<dc:creator>Hastie, A.</dc:creator>
<dc:creator>Ortega, V.</dc:creator>
<dc:creator>Lappalainen, T.</dc:creator>
<dc:creator>Christenson, S.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Cho, M.</dc:creator>
<dc:creator>Kesimer, M.</dc:creator>
<dc:creator>Kelada, S.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.28.509838</dc:identifier>
<dc:title><![CDATA[Genetic Regulators of Sputum Mucin Concentration and Their Associations with COPD Phenotypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.28.509882v1?rss=1">
<title>
<![CDATA[
A host defense peptide mimetic, brilacidin, potentiates caspofungin antifungal activity against human pathogenic fungi 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.28.509882v1?rss=1</link>
<description><![CDATA[
A. fumigatus is the main etiological agent of a group of heterogeneous diseases called aspergillosis of which the most lethal form is the invasive pulmonary aspergillosis (IPA). Fungicidal azoles and amphotericin B are the first line defense against A. fumigatus, but fungistatic echinocandins, such as caspofungin (CAS), can be used as salvage therapy for IPA. Here, we screened repurposing libraries and identified several compounds that potentiate CAS activity against A. fumigatus, including the host defense peptide mimetic, brilacidin (BRI). BRI converts CAS into a fungicidal drug and potentiates voriconazole (VOR) against A. fumigatus. BRI increases the ability of both CAS and VOR to control A. fumigatus biofilm growth. BRI depolarizes the A. fumigatus cell membrane leading to disruption of membrane potential. By using a combination of protein kinase inhibitors and screening of a catalytic subunit null mutant library, we identified the mitogen activated protein kinase (MAPK) MpkA and the phosphatase calcineurin as mediators of the synergistic action of BRI. These results suggest the most likely BRI mechanism of action for CAS potentiation is the inhibition of A. fumigatus cell wall integrity (CWI) pathway. BRI potentiates CAS activity against C. albicans, C. auris, and C. neoformans. Interestingly, BRI overcomes the CAS-acquired resistance in both A. fumigatus and C. albicans and the CAS-intrinsic resistance in C. neoformans. BRI also has an additive effect on the activity of posaconazole (POSA) against several Mucorales fungi. Cell toxicity assays and fungal burden studies in an immunosuppressed murine model of IPA showed that BRI combined with CAS is not toxic to the cells and significantly clears A. fumigatus lung infection, respectively. Our results indicate that combinations of BRI and antifungal drugs in clinical use are likely to improve the treatment outcome of IPA and other fungal infections.
]]></description>
<dc:creator>dos Reis, T. F.</dc:creator>
<dc:creator>de Castro, P. A.</dc:creator>
<dc:creator>Bastos, R.</dc:creator>
<dc:creator>Pinzan, C. F.</dc:creator>
<dc:creator>Souza, P. F.</dc:creator>
<dc:creator>Ackloo, S.</dc:creator>
<dc:creator>Hossain, M. A.</dc:creator>
<dc:creator>Drewry, D.</dc:creator>
<dc:creator>Alkhazraji, S.</dc:creator>
<dc:creator>Ibrahim, A.</dc:creator>
<dc:creator>Jo, H.</dc:creator>
<dc:creator>deGrado, W. F.</dc:creator>
<dc:creator>Goldman, G.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.28.509882</dc:identifier>
<dc:title><![CDATA[A host defense peptide mimetic, brilacidin, potentiates caspofungin antifungal activity against human pathogenic fungi]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.28.509901v1?rss=1">
<title>
<![CDATA[
DASH/Dam1 complex mutants stabilize ploidy by weakening kinetochore-microtubule attachments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.28.509901v1?rss=1</link>
<description><![CDATA[
Forcing budding yeast to chromatinize their DNA with human histones manifests an abrupt fitness cost. We previously proposed chromosomal aneuploidy and missense mutations as two potential modes of adaptation to histone humanization. Here we show that aneuploidy in histone-humanized yeasts is specific to a subset of chromosomes, defined by their centromeric evolutionary origins, however, they are not adaptive. Instead we show that a set of missense mutations in outer kineto-chore proteins drive adaptation to human histones. Further, we characterize the molecular mechanism of two mutants of the outer kinetochore DASH/Dam1 complex, which reduce aneuploidy by suppression of chromosome instability. Molecular modeling and biochemical experiments show that these two mutants likely disrupt a conserved oligomerization interface thereby weakening microtubule attachments. Lastly, we show that one mutant, DAD1E50D, while suppressing chromosome instability in mitosis, leads to gross defects in meiosis. In sum, our data show how a set of point mutations evolved in the histone-humanized yeasts to counterbalance human histone induced chromosomal instability through weakening microtubule interactions, eventually promoting a return to euploidy.
]]></description>
<dc:creator>Haase, M. A. B.</dc:creator>
<dc:creator>Olafsson, G.</dc:creator>
<dc:creator>Flores, R. L.</dc:creator>
<dc:creator>Boakye-Ansah, E.</dc:creator>
<dc:creator>Zelter, A.</dc:creator>
<dc:creator>Dickinson, M. S.</dc:creator>
<dc:creator>Stefanita, L. L.</dc:creator>
<dc:creator>Truong, D. M.</dc:creator>
<dc:creator>Asbury, C. L.</dc:creator>
<dc:creator>Davis, T. N.</dc:creator>
<dc:creator>Boeke, J. D.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.28.509901</dc:identifier>
<dc:title><![CDATA[DASH/Dam1 complex mutants stabilize ploidy by weakening kinetochore-microtubule attachments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.28.510016v1?rss=1">
<title>
<![CDATA[
De novo protein identification in mammalian sperm using high-resolution in situ cryo-electron tomography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.28.510016v1?rss=1</link>
<description><![CDATA[
Understanding molecular mechanisms of cellular pathways requires knowledge of the identities of participating proteins, their cellular localization and their 3D structures. Contemporary workflows typically require multiple techniques to identify target proteins, track their localization using fluorescence microscopy, followed by in vitro structure determination. To identify mammal-specific sperm proteins and understand their functions, we developed a visual proteomics workflow to directly address these challenges. Our in situ cryo-electron tomography and subtomogram averaging provided 6.0 [A] resolution reconstructions of axonemal microtubules and their associated proteins. The well-resolved secondary and tertiary structures allowed us to computationally match, in an unbiased manner, novel densities in our 3D reconstruction maps with 21,615 AlphaFold2-predicted protein models of the mouse proteome. We identified Tektin 5, CCDC105 and SPACA9 as novel microtubule inner proteins that form an extensive network crosslinking the lumen of microtubule and existing proteins. Additional biochemical and mass spectrometry analyses helped validate potential candidates. The novel axonemal sperm structures identified by this approach form an extensive interaction network within the lumen of microtubules, suggesting they have a role in the mechanical and elastic properties of the microtubule filaments required for the vigorous beating motions of flagella.
]]></description>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Shiozaki, M.</dc:creator>
<dc:creator>Haas, K. M.</dc:creator>
<dc:creator>Zhao, S.</dc:creator>
<dc:creator>Guo, C.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Kaake, R. M.</dc:creator>
<dc:creator>Vale, R. D.</dc:creator>
<dc:creator>Agard, D. A.</dc:creator>
<dc:date>2022-09-29</dc:date>
<dc:identifier>doi:10.1101/2022.09.28.510016</dc:identifier>
<dc:title><![CDATA[De novo protein identification in mammalian sperm using high-resolution in situ cryo-electron tomography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.28.506941v1?rss=1">
<title>
<![CDATA[
A tradeoff between enterovirus A71 particle stability and cell entry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.28.506941v1?rss=1</link>
<description><![CDATA[
A central role of viral capsids is to protect the viral genome from the harsh extracellular environment while facilitating initiation of infection when the virus encounters a target cell. Viruses are thought to have evolved an optimal equilibrium between particle stability and efficiency of cell entry. In this study, we genetically perturbed this equilibrium in a non-enveloped virus, enterovirus A71 to determine its structural basis. We isolated a single-point mutation variant with increased particle thermotolerance and decreased efficiency of cell entry. Using cryo-electron microscopy and molecular dynamics simulations, we determined that the thermostable native particles have acquired an expanded conformation that results in a significant increase in protein dynamics. Examining the uncoating intermediate states of the thermostable variant suggests a pathway, where the lipid pocket factor is released first, followed by internal VP4 and finally the viral RNA.
]]></description>
<dc:creator>Catching, A.</dc:creator>
<dc:creator>Yeh, M. T.</dc:creator>
<dc:creator>Bianco, S.</dc:creator>
<dc:creator>Capponi, S.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:date>2022-09-30</dc:date>
<dc:identifier>doi:10.1101/2022.09.28.506941</dc:identifier>
<dc:title><![CDATA[A tradeoff between enterovirus A71 particle stability and cell entry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.28.509955v1?rss=1">
<title>
<![CDATA[
Single-cell sequencing identifies novel atypical B cell subsets with distinct effector functions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.28.509955v1?rss=1</link>
<description><![CDATA[
Atypical B cells are a population of activated B cells that are commonly enriched in individuals with chronic immune activation, but are also part of a normal immune response to infection or vaccination. Prior studies to determine the function of these cells have yielded conflicting results, possibly due to functional heterogeneity among this B cell population. To better define the role(s) of atypical B cells in the host adaptive immune response, we performed single-cell sequencing of transcriptomes, cell surface markers, and B cell receptors in individuals with chronic Plasmodium falciparum exposure, a condition known to lead to accumulation of circulating atypical B cells. Our studies identified three previously uncharacterized populations of atypical B cells with distinct transcriptional and functional profiles, that separate into two differentiation pathways. We identified a set of cell surface markers to distinguish these atypical B cell subsets and confirmed their presence in malaria-experienced children and adults using flow cytometry. Plasmodium falciparum-specific cells were present in equal proportions within each of these atypical B cell populations, indicating that all three subsets develop in response to antigen stimulation. However, we observed marked differences among the three subsets in their ability to produce IgG upon T-cell-dependent activation. Collectively, our findings help explain the conflicting observations in prior studies on the functions of atypical B cells and provide a better understanding of their role in the adaptive immune response in chronic inflammatory conditions.

One sentence summaryAtypical B cells consist of three subsets that may play distinct roles in the host adaptive immune response.
]]></description>
<dc:creator>Reyes, R. A.</dc:creator>
<dc:creator>Batugedara, G.</dc:creator>
<dc:creator>Dutta, P.</dc:creator>
<dc:creator>Reers, A.</dc:creator>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Ssewanyana, I.</dc:creator>
<dc:creator>Feeney, M. E.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:creator>Bol, S.</dc:creator>
<dc:creator>Bunnik, E. M.</dc:creator>
<dc:date>2022-09-30</dc:date>
<dc:identifier>doi:10.1101/2022.09.28.509955</dc:identifier>
<dc:title><![CDATA[Single-cell sequencing identifies novel atypical B cell subsets with distinct effector functions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.29.509997v1?rss=1">
<title>
<![CDATA[
mTOR inhibitor-independent Autophagy Activator Ameliorates Cellular Tauopathy and Prionopathy Neurodegeneration Phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.29.509997v1?rss=1</link>
<description><![CDATA[
Autophagy-lysosomal impairment is an early and prominent feature of neurodegeneration. Autophagy activation reduces protein aggregates and lipid level abnormalities. We performed a high-content imaging-based screen assessing 940,000 small molecules to identify those that reduce lipid droplet numbers. Of 77 validated, structurally diverse hits, 24 increased autophagy flux reporter activity, consistent with accelerated lipid droplet clearance by lipophagy. Of these, we show that CCT020312 activates autophagy independently of mammalian target of rapamycin (mTOR) inhibition, to avoid immunosuppression. CCT020312 reduced insoluble phosphorylated tau levels and tau-mediated neuronal stress vulnerability, as well as reducing intracellular A{beta} levels within directly induced neurons bearing epigenetic marks of aging derived from Alzheimers patient fibroblasts. Moreover, CCT020312 cleared mutant prion protein aggregates and normalized trafficking deficiencies in axons of a cellular model of familial prion disease. Autophagy is widely considered a promising strategy to attenuate neurodegeneration, and here we introduce a strategy to discover new pharmacology.
]]></description>
<dc:creator>Yoon, L.</dc:creator>
<dc:creator>Botham, R. C.</dc:creator>
<dc:creator>Verhelle, A.</dc:creator>
<dc:creator>Cole, C.</dc:creator>
<dc:creator>Tan, E. P.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Sanz-Martinez, P.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Cuoco, C. A.</dc:creator>
<dc:creator>Chou, C. C.</dc:creator>
<dc:creator>Labra, S.</dc:creator>
<dc:creator>Elia, L. P.</dc:creator>
<dc:creator>Massey, L. A.</dc:creator>
<dc:creator>Lee, K.</dc:creator>
<dc:creator>Xiao, Q.</dc:creator>
<dc:creator>Kline, G. M.</dc:creator>
<dc:creator>Ta, A.</dc:creator>
<dc:creator>Ardejani, M. S.</dc:creator>
<dc:creator>Cano-Franco, S.</dc:creator>
<dc:creator>Lyang, N. J.</dc:creator>
<dc:creator>Hou, W. C.</dc:creator>
<dc:creator>Yu, A.</dc:creator>
<dc:creator>Fox, S.</dc:creator>
<dc:creator>Ko, Y.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Wulkop-Gil, C.</dc:creator>
<dc:creator>Ibrahim, L. H.</dc:creator>
<dc:creator>Jiang, S.</dc:creator>
<dc:creator>Meneses, A.</dc:creator>
<dc:creator>Nelson, L. T.</dc:creator>
<dc:creator>Peng, H.</dc:creator>
<dc:creator>Lipton, S. A.</dc:creator>
<dc:creator>Bollong, M. J.</dc:creator>
<dc:creator>Hansen, M.</dc:creator>
<dc:creator>Morimoto, R. I.</dc:creator>
<dc:creator>Finley, D.</dc:creator>
<dc:creator>Petrassi, H. M.</dc:creator>
<dc:creator>Wiseman, R. L.</dc:creator>
<dc:creator>Powers, E. T.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:creator>Garza, D.</dc:creator>
<dc:creator>Prado, M. A.</dc:creator>
<dc:creator>Dikic, I.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:creator>Johnson, K. A.</dc:creator>
<dc:creator>Silva, M. C.</dc:creator>
<dc:creator>Haggar</dc:creator>
<dc:date>2022-09-30</dc:date>
<dc:identifier>doi:10.1101/2022.09.29.509997</dc:identifier>
<dc:title><![CDATA[mTOR inhibitor-independent Autophagy Activator Ameliorates Cellular Tauopathy and Prionopathy Neurodegeneration Phenotypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.29.510156v1?rss=1">
<title>
<![CDATA[
Role of intraflagellar transport in transcriptional control during flagellar regeneration in Chlamydomonas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.29.510156v1?rss=1</link>
<description><![CDATA[
Biosynthesis of organelle precursors is a central part of the organelle size control problem, but what systems are required to control precursor production? Genes encoding flagellar proteins are upregulated during flagellar regeneration in Chlamydomonas, and this upregulation is critical for flagella to reach their final length, but it not known how the cell triggers these genes during regeneration. Here we present two models based on transcriptional repressor that is either produced in the flagellum, or else is produced in the cell body and sequestered in the growing flagellum. We show that both models lead to stable flagellar length control, can reproduce the observed dynamics of gene expression, and are consistent with the effects of protein synthesis inhibitors on gene expression. The two models make opposite predictions regarding the effect of mutations that block intraflagellar transport (IFT). Using quantitative measurements of gene expression, we show that gene expression during flagellar regeneration is greatly reduced in mutations of FLA3, FLA8, and FLA10, which encode the three components of the heterotrimer kinesin-2 that drives IFT. This result is consistent with the predictions of the model in which a repressor is sequestered in the flagellum by IFT. In contrast to the effects of IFT mutants, we find that inhibiting axonemal assembly has much less effect on gene expression, suggesting that transport is more important than axonemal assembly. The repressor sequestration model allows precursor production to occur when flagella are growing rapidly, representing a form of derivative control.
]]></description>
<dc:creator>Perlaza, K.</dc:creator>
<dc:creator>Zamora, I.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2022-09-30</dc:date>
<dc:identifier>doi:10.1101/2022.09.29.510156</dc:identifier>
<dc:title><![CDATA[Role of intraflagellar transport in transcriptional control during flagellar regeneration in Chlamydomonas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.30.510175v1?rss=1">
<title>
<![CDATA[
Stacked binding of a small molecule PET tracer to Alzheimer's tau paired helical filaments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.30.510175v1?rss=1</link>
<description><![CDATA[
Neurodegenerative diseases (NDs) are characterized by the formation of amyloid filaments that adopt disease-specific conformations in the brain. Recently developed small molecules hold promise as diagnostics and possible therapeutics for NDs, but their binding mechanisms to amyloid filaments remain unknown. Here, we used cryo-electron microscopy (cryo-EM) to determine a 2.7 [A] structure of Alzheimers disease patient-derived tau paired-helical filaments incubated with the GTP-1 PET probe. GTP-1 is bound stoichiometrically along an exposed cleft of each protofilament in a stacked arrangement that matches the fibrils symmetry. Multiscale modeling revealed favorable pi-pi aromatic stacking interactions between GTP-1 molecules that, together with small molecule-protein contacts, result in high affinity binding. This binding mode offers new insight into designing compounds for diagnosis and treatment of specific NDs.

One Sentence SummaryCryo-EM structure reveals a novel stacked arrangement of the GTP-1 PET ligand bound to Alzheimers disease tau filaments.
]]></description>
<dc:creator>Merz, G. E.</dc:creator>
<dc:creator>Chalkley, M. J.</dc:creator>
<dc:creator>Tan, S. K.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Prusiner, S. B.</dc:creator>
<dc:creator>Paras, N. A.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:creator>Southworth, D. R.</dc:creator>
<dc:date>2022-10-03</dc:date>
<dc:identifier>doi:10.1101/2022.09.30.510175</dc:identifier>
<dc:title><![CDATA[Stacked binding of a small molecule PET tracer to Alzheimer's tau paired helical filaments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.02.510349v1?rss=1">
<title>
<![CDATA[
Pharmacologic Inhibition of DYRK1A Results in MYC Hyperactivation and ERK Hyperphosphorylation rendering KMT2A-R ALL Cells Sensitive to BCL2 Inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.02.510349v1?rss=1</link>
<description><![CDATA[
KMT2A-rearranged (KMT2A-R) B cell acute lymphoblastic leukemia (ALL) is a high-risk disease in children and adults that is often chemotherapy resistant. To identify non-cytotoxic approaches to therapy, we performed a domain-specific kinome-wide CRISPR screen in KMT2A-R cell lines and patient derived xenograft samples (PDX) and identified dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) as a potential target. Pharmacologic inhibition of the KMT2A-fusion transcriptional co-regulator Menin released the KMT2A-fusion complex from the DYRK1A promoter thereby lowering DYRK1A expression levels confirming DYRK1A as a direct target of the KMT2A fusion oncogene. Direct pharmacologic inhibition of DYRK1A decreased cell proliferation of KMT2A-R ALL, thereby confirming the requirement of DYRK1A in this ALL subtype. To further understand the biologic function of DYRK1A in KMT2A-R ALL, we leveraged pharmacologic DYRK1A inhibitors in KMT2A-R PDX and cell line models. DYRK1A inhibition consistently led to upregulation of MYC protein levels, and hyperphosphorylation of ERK, which we confirmed via in vivo treatment experiments. Furthermore, DYRK1A inhibition decreased ALL burden in mice. Our results further demonstrate that DYRK1A inhibition induces the proapoptotic factor BIM, but ERK hyperphosphorylation is the driving event that induces cell cycle arrest. In contrast, combined treatment of KMT2A-R ALL cells in vitro and in vivo with DYRK1A inhibitors and the BCL2 inhibitor, venetoclax, synergistically decreases cell survival and reduced the leukemic burden in mice. Taken together these results demonstrate a unique function of DYRK1A specially in KMT2A-R ALL. Synergistic inhibition of DRYK1A and BCL2 may provide a low-toxic approach to treat this high risk ALL subtype.
]]></description>
<dc:creator>Hurtz, C.</dc:creator>
<dc:creator>Ayyadevara, V. S. S. A.</dc:creator>
<dc:creator>Wertheim, G.</dc:creator>
<dc:creator>Chukinas, J.</dc:creator>
<dc:creator>Loftus, J. P.</dc:creator>
<dc:creator>Lee, S. J.</dc:creator>
<dc:creator>Kumar, A.</dc:creator>
<dc:creator>Bhansali, R. S.</dc:creator>
<dc:creator>Swaminathan, S.</dc:creator>
<dc:creator>Geng, H.</dc:creator>
<dc:creator>Milne, T.</dc:creator>
<dc:creator>Hua, X.</dc:creator>
<dc:creator>Bernt, K. M.</dc:creator>
<dc:creator>Besson, T.</dc:creator>
<dc:creator>Shi, J.</dc:creator>
<dc:creator>Crispino, J. D.</dc:creator>
<dc:creator>Carroll, M.</dc:creator>
<dc:creator>Tasian, S. K.</dc:creator>
<dc:date>2022-10-04</dc:date>
<dc:identifier>doi:10.1101/2022.10.02.510349</dc:identifier>
<dc:title><![CDATA[Pharmacologic Inhibition of DYRK1A Results in MYC Hyperactivation and ERK Hyperphosphorylation rendering KMT2A-R ALL Cells Sensitive to BCL2 Inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.02.510551v1?rss=1">
<title>
<![CDATA[
Proteomic analysis of flagella from Chlamydomonas mutants lacking the central pair apparatus reveals loss of radial spoke head proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.02.510551v1?rss=1</link>
<description><![CDATA[
The flagellar axoneme is a complex protein-based machine capable of generating motile forces by coordinating the action of thousands of dynein motors. A key element of the axoneme is the central pair apparatus, consisting of a pair of microtubules surrounded by additional structures. In an effort to better understand the organization of the central pair, we used 2D DIGE to identify proteins that are depleted from flagella isolated from two different Chlamydomonas reinhardtii mutants, pf15 and pf18, that lack the central pair. The set of proteins contained almost no components of the central apparatus. We find that three proteins of the radial spoke head RSP1, RSP9, and RSP10, as well as a number of other protein components associated with the outer doublets, are depleted from flagella of mutants lacking the central apparatus. Two of the other proteins depleted from pf15 and pf18 flagella, the microtubule inner proteins (MIPs) FAP21 and FAP161, are missing from the genome of Thalassiosira, an organism that lacks a central pair and radial spokes, and RNAi of FAP21 in planaria shows that it has a role in ciliary motility. Based on the depletion of radial spoke head proteins, as well as MIPs and other axonemal components, from flagella lacking the central pair apparatus, we hypothesize that the central apparatus may play a role in scaffolding the assembly or retention of radial spokes and other axonemal structures.
]]></description>
<dc:creator>Wemmer, K. A.</dc:creator>
<dc:creator>AZIMZADEH, J. F.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2022-10-04</dc:date>
<dc:identifier>doi:10.1101/2022.10.02.510551</dc:identifier>
<dc:title><![CDATA[Proteomic analysis of flagella from Chlamydomonas mutants lacking the central pair apparatus reveals loss of radial spoke head proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.03.510592v1?rss=1">
<title>
<![CDATA[
The cellular states and fates of shed intestinal cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.03.510592v1?rss=1</link>
<description><![CDATA[
The intestinal epithelium is replaced every few days1. Enterocytes are shed into the gut lumen predominantly from the tips of villi3,4, and are believed to rapidly die upon their dissociation from the tissue. However, technical limitations prohibited studying the cellular states and fates of shed intestinal cells. Here, we used bulk and single cell RNA sequencing of mouse intestinal fecal washes to demonstrate that shed epithelial cells remain viable and up-regulate distinct anti-microbial programs upon shedding. We further identify abundant shedding of immune cells, a process that is elevated in DSS-induced colitis. We find that fecal host transcriptomics mirrors changes in the intestinal tissue following perturbations. Our study suggests potential functions of shed cells in the intestinal lumen and demonstrates that host cell transcriptomes in intestinal washes can be used to probe tissue states.
]]></description>
<dc:creator>Bahar Halpern, K.</dc:creator>
<dc:creator>Biram, A.</dc:creator>
<dc:creator>Egozi, A.</dc:creator>
<dc:creator>Korem Kohanim, Y.</dc:creator>
<dc:creator>Shulman, Z.</dc:creator>
<dc:creator>Itzkovitz, S.</dc:creator>
<dc:date>2022-10-04</dc:date>
<dc:identifier>doi:10.1101/2022.10.03.510592</dc:identifier>
<dc:title><![CDATA[The cellular states and fates of shed intestinal cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.03.510667v1?rss=1">
<title>
<![CDATA[
EMC holdase:CaV1.2/CaVβ3 complex and CaV1.2 channel structures reveal CaV assembly and drug binding mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.03.510667v1?rss=1</link>
<description><![CDATA[
Voltage-gated ion channels (VGICs) comprise multiple structural units whose assembly is required for function1,2. There is scant structural understanding of how VGIC subunits assemble and whether chaperone proteins are required. High-voltage activated calcium channels (CaVs)3,4 are paradigmatic multi-subunit VGICs from electrically excitable tissues whose function and trafficking is powerfully shaped by interactions between pore-forming CaV1 or CaV2 CaV13 and auxiliary CaV{beta}5, and CaV2{delta} subunits6,7. Here, we present cryo-EM structures of human brain and cardiac CaV1.2 bound with CaV{beta}3 to a chaperone, the endoplasmic reticulum membrane protein complex (EMC)8,9, and of the isolated CaV1.2/CaV{beta}3/CaV2{delta}-1 channel. These provide an unprecedented view of an EMC holdase:client complex and define EMC sites, the TM and Cyto docks, whose interaction with the client channel cause partial extraction of a pore subunit and splay open the CaV2{delta} interaction site. The structures further identify the CaV2{delta} binding site for gabapentinoid anti-pain and anti-anxiety drugs6, show that EMC and CaV2{delta} channel interactions are mutually exclusive, and indicate that EMC to CaV2{delta} handoff involves a Ca2+-dependent step and ordering of multiple CaV1.2 elements. Together, the structures unveil a CaV assembly intermediate and previously unknown EMC client binding sites that have broad implications for biogenesis of VGICs and other membrane proteins.
]]></description>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Mondal, A.</dc:creator>
<dc:creator>Abderemane-Ali, F.</dc:creator>
<dc:creator>Montano, J.</dc:creator>
<dc:creator>Zaro, B.</dc:creator>
<dc:creator>Minor, D. L.</dc:creator>
<dc:date>2022-10-05</dc:date>
<dc:identifier>doi:10.1101/2022.10.03.510667</dc:identifier>
<dc:title><![CDATA[EMC holdase:CaV1.2/CaVβ3 complex and CaV1.2 channel structures reveal CaV assembly and drug binding mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.04.510859v1?rss=1">
<title>
<![CDATA[
Three-dimensional genome re-wiring in loci with Human Accelerated Regions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.04.510859v1?rss=1</link>
<description><![CDATA[
Human Accelerated Regions (HARs) are conserved genomic loci that evolved at an accelerated rate in the human lineage and may underlie human-specific traits. We generated HARs and chimpanzee accelerated regions with the largest alignment of mammalian genomes to date. To facilitate exploration of accelerated evolution in other lineages, we implemented an open-source Nextflow pipeline that runs on any computing platform. Combining deep-learning with chromatin capture experiments in human and chimpanzee neural progenitor cells, we discovered a significant enrichment of HARs in topologically associating domains (TADs) containing human-specific genomic variants that change three-dimensional (3D) genome organization. Differential gene expression between humans and chimpanzees at these loci in multiple cell types suggests rewiring of regulatory interactions between HARs and neurodevelopmental genes. Thus, comparative genomics together with models of 3D genome folding revealed enhancer hijacking as an explanation for the rapid evolution of HARs.

One-Sentence SummaryHuman-specific changes to 3D genome organization may have contributed to rapid evolution of mammalian-conserved loci in the human genome.
]]></description>
<dc:creator>Keough, K. C.</dc:creator>
<dc:creator>Whalen, S.</dc:creator>
<dc:creator>Inoue, F.</dc:creator>
<dc:creator>Przytycki, P. F.</dc:creator>
<dc:creator>Fair, T.</dc:creator>
<dc:creator>Deng, C.</dc:creator>
<dc:creator>Steyert, M.</dc:creator>
<dc:creator>Ryu, H.</dc:creator>
<dc:creator>Lindblad-Toh, K.</dc:creator>
<dc:creator>Karlsson, E.</dc:creator>
<dc:creator>Zoonomia Consortium,</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Ahituv, N. J.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2022-10-05</dc:date>
<dc:identifier>doi:10.1101/2022.10.04.510859</dc:identifier>
<dc:title><![CDATA[Three-dimensional genome re-wiring in loci with Human Accelerated Regions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.05.511009v1?rss=1">
<title>
<![CDATA[
Opposing transcription factors MYCL and HEY1 mediate the Notch-dependent airway stem cell fate decision 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.05.511009v1?rss=1</link>
<description><![CDATA[
Tissue function depends on the relative proportions of multiple cell types. In the airway, basal stem cells differentiate into both multiciliated and secretory cells, which together protect the lungs from inhaled pathogens and particulates. To define how airway stem cells differentiate, we mapped differentiation trajectories using single-cell mRNA sequencing (scRNA-seq) and identified a transitional intermediate cell state in between basal stem cells and differentiated cells. These intermediate cells induce different gene expression programs that precede differentiation into either multiciliated or secretory cells. For example, we found that within the intermediate cell population, multiciliated cell precursors express Mycl, encoding a MYC-family transcription factor, and secretory cell precursors express Hey1, encoding a transcriptional repressor. We also found that Notch signaling acts on intermediate cells to repress Mycl and induce Hey1. We further show MYCL expression is sufficient to drive multiciliated cell fate, whereas HEY1 expression is sufficient to repress multiciliated cell fate. Using CUT&RUN, we made the surprising observation that MYCL and HEY1 bind to many of the same regulatory elements near genes encoding early regulators of multiciliated cell differentiation. We conclude that intermediate cells receiving Notch signals induce HEY1 to repress the multiciliated cell fate and become secretory cells, while intermediate cells not receiving Notch signals induce MYCL to promote the multiciliated cell fate. These experiments reveal that during airway stem cell differentiation Notch signaling balances the production of two different cell types by regulating the functions of two opposing transcription factors, MYCL and HEY1.
]]></description>
<dc:creator>Byrnes, L. E.</dc:creator>
<dc:creator>Deleon, R.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:creator>Choksi, S. P.</dc:creator>
<dc:date>2022-10-05</dc:date>
<dc:identifier>doi:10.1101/2022.10.05.511009</dc:identifier>
<dc:title><![CDATA[Opposing transcription factors MYCL and HEY1 mediate the Notch-dependent airway stem cell fate decision]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.05.511060v1?rss=1">
<title>
<![CDATA[
Circularization of rv0678 for genotypic bedaquiline resistance testing of Mycobacterium tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.05.511060v1?rss=1</link>
<description><![CDATA[
Circular DNA offers benefits over linear DNA in diagnostic and field assays, but currently, circular DNA generation is lengthy, inefficient, highly dependent on the length and sequence of DNA, and can result in unwanted chimeras. We present streamlined methods for generating PCR-targeted circular DNA from a 700bp amplicon of rv0678, the high GC content (65%) gene implicated in Mycobacterium tuberculosis bedaquiline resistance, and demonstrate that these methods work as desired. We employ self-circularization with and without splints, a Gibson cloning-based approach, and novel two novel methods for generating pseudocircular DNA. The circular DNA can be used as a template for rolling circle PCR followed by long-read sequencing, allowing for the error correction of sequence data, and improving the confidence in the drug resistance determination and strain identification; and ultimately improving patient treatment.
]]></description>
<dc:creator>Limberis, J.</dc:creator>
<dc:creator>Nalyvayko, A.</dc:creator>
<dc:creator>Ernst, J.</dc:creator>
<dc:creator>Metcalfe, J. Z.</dc:creator>
<dc:date>2022-10-06</dc:date>
<dc:identifier>doi:10.1101/2022.10.05.511060</dc:identifier>
<dc:title><![CDATA[Circularization of rv0678 for genotypic bedaquiline resistance testing of Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.07.511350v1?rss=1">
<title>
<![CDATA[
Epigenomic and chromosomal architectural reconfiguration in developing human frontal cortex and hippocampus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.07.511350v1?rss=1</link>
<description><![CDATA[
The human frontal cortex and hippocampus play critical roles in learning and cognition. We investigated the epigenomic and 3D chromatin conformational reorganization during the development of the frontal cortex and hippocampus, using more than 53,000 joint single-nucleus profiles of chromatin conformation and DNA methylation (sn-m3C-seq). The remodeling of DNA methylation predominantly occurs during late-gestational to early-infant development and is temporally separated from chromatin conformation dynamics. Neurons have a unique Domain-Dominant chromatin conformation that is different from the Compartment-Dominant conformation of glial cells and non-brain tissues. We reconstructed the regulatory programs of cell-type differentiation and found putatively causal common variants for schizophrenia strongly overlap with chromatin loop-connected, cell-type-specific regulatory regions. Our data demonstrate that single-cell 3D-regulome is an effective approach for dissecting neuropsychiatric risk loci.
]]></description>
<dc:creator>Heffel, M. G.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Lee, D.-S.</dc:creator>
<dc:creator>Hou, K.</dc:creator>
<dc:creator>Pastor Alonso, O.</dc:creator>
<dc:creator>Abuhanna, K.</dc:creator>
<dc:creator>Schmitt, A. D.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Haeussler, M.</dc:creator>
<dc:creator>Wick, B.</dc:creator>
<dc:creator>Zhang, M. J.</dc:creator>
<dc:creator>Xie, F.</dc:creator>
<dc:creator>Ziffra, R. S.</dc:creator>
<dc:creator>Mukamel, E. A.</dc:creator>
<dc:creator>Eskin, E.</dc:creator>
<dc:creator>Pasaniuc, B.</dc:creator>
<dc:creator>Ecker, J. R.</dc:creator>
<dc:creator>Dixon, J.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Paredes, M. F.</dc:creator>
<dc:creator>Luo, C.</dc:creator>
<dc:date>2022-10-08</dc:date>
<dc:identifier>doi:10.1101/2022.10.07.511350</dc:identifier>
<dc:title><![CDATA[Epigenomic and chromosomal architectural reconfiguration in developing human frontal cortex and hippocampus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.08.511444v1?rss=1">
<title>
<![CDATA[
An unbiased screen identified the Hsp70-BAG3 complex as a regulator of myosin binding protein C3 stability. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.08.511444v1?rss=1</link>
<description><![CDATA[
Structured AbstractO_ST_ABSObjectiveC_ST_ABSWe aim to identify regulators of myosin binding protein C3 (MyBP-C) protein homeostasis.

BackgroundVariants in myosin binding protein C3 (MYBPC3) account for approximately 50% of familial hypertrophic cardiomyopathy (HCM). Most pathogenic variants in MYBPC3 are truncating variants that lead to reduced total levels of MyBP-C protein. Elucidation of the pathways that regulate MyBP-C protein homeostasis could uncover new therapeutic strategies that restore normal protein levels.

MethodWe developed a high-throughput screen to identify compounds that can increase or decrease steady-state levels of MyBP-C in an induced pluripotent stem cell cardiomyocyte (iPSC-CM) model derived from a patient with HCM. To normalize results, we also monitored effects on myosin heavy chain (MYH) and focused on those molecules that selectively modulated MyBP-C levels.

ResultsScreening a library of 2,426 known biologically active compounds, we identified compounds which either decreased (241/2426, 9.9%) or increased (29/2426, 1.2%) MyBP-C/MYH levels. After a rigorous validation process, including a counter screen for cellular toxicity, two compounds (JG98 and parthenolide) were confirmed as decreasing MyBP-C levels and no compounds were confirmed to increase MyBP-C levels. For further studies, we focused on JG98, which is an allosteric modulator of heat shock protein 70 (Hsp70), inhibiting its interaction with BAG domain co-chaperones. We found that genetic reduction of BAG3 phenocopies treatment with JG98 by reducing MyBP-C protein levels.

ConclusionAn unbiased compound screen identified the Hsp70-BAG3 complex as a regulator of MyBP-C stability. Thus, approaches that stimulate this complexs function may be beneficial in the treatment of HCM.

Highlights- Hypertrophic cardiomyopathy (HCM) is commonly caused by pathogenic MYBPC3 variants that reduce total wild-type MyBP-C (the protein encoded by MYBPC3).
- It is critical to understand the regulators of MyBP-C protein homeostasis to uncover novel therapeutic strategies.
- We developed and executed a high-throughput chemical screen in iPSC-CMs to identify compounds which alter steady-state levels of MyBP-C protein, revealing two compounds, JG98 and parthenolide, that significantly reduced MyBP-C levels.
- Validation studies suggest that the complex between heat shock protein 70 (Hsp70) and its co-chaperone BAG3 is a dynamic regulator of MyBP-C stability, suggesting that this axis could be a new therapeutic target for HCM.
]]></description>
<dc:creator>Thompson, A. D.</dc:creator>
<dc:creator>Wagner, M. J.</dc:creator>
<dc:creator>Rodriguez, J.</dc:creator>
<dc:creator>Malhotra, A.</dc:creator>
<dc:creator>Vander Roest, S.</dc:creator>
<dc:creator>Lilienthal, U.</dc:creator>
<dc:creator>Shao, H.</dc:creator>
<dc:creator>Yob, J.</dc:creator>
<dc:creator>Prosser, B. L.</dc:creator>
<dc:creator>Helms, A. S.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Ginsburg, D. E.</dc:creator>
<dc:creator>Day, S. M.</dc:creator>
<dc:date>2022-10-08</dc:date>
<dc:identifier>doi:10.1101/2022.10.08.511444</dc:identifier>
<dc:title><![CDATA[An unbiased screen identified the Hsp70-BAG3 complex as a regulator of myosin binding protein C3 stability.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.09.511478v1?rss=1">
<title>
<![CDATA[
Structural Genomics of the Human Dopamine Receptor System 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.09.511478v1?rss=1</link>
<description><![CDATA[
The dopamine system, including five dopamine receptors (D1R to D5R), plays essential roles in the central nervous system (CNS) and ligands that activate dopamine receptors have been used to treat many neuropsychiatric disorders, including Parkinsons Disease (PD) and schizophrenia. Here, we report five cryo-EM structures of all subtypes of human dopamine receptors in complex with G-protein and bound to the pan agonist, Rotigotine, which is used to treat PD and restless legs syndrome. The structures reveal the basis of Rotigotine binding modes to different dopamine receptors. Structural analysis together with functional assays illuminate determinants of ligand polypharmacology and selectivity. The structures also uncover the mechanisms of the dopamine receptor activation, unique structural features among the five receptor subtypes, and the basis of G-protein coupling specificity. Our works provide a comprehensive set of structural templates for the rational design of specific ligands to treat CNS diseases targeting the dopaminergic system.
]]></description>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Krumm, B. E.</dc:creator>
<dc:creator>Zhuang, Y.</dc:creator>
<dc:creator>Mao, C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Huang, X.-P.</dc:creator>
<dc:creator>Liu, Y.-F.</dc:creator>
<dc:creator>He, X.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Yin, W.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Roth, B. L.</dc:creator>
<dc:creator>Xu, H. E.</dc:creator>
<dc:date>2022-10-10</dc:date>
<dc:identifier>doi:10.1101/2022.10.09.511478</dc:identifier>
<dc:title><![CDATA[Structural Genomics of the Human Dopamine Receptor System]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.10.511511v1?rss=1">
<title>
<![CDATA[
Structural surfaceomics reveals an AML-specific conformation of Integrin-β2 as a CAR-T therapy target 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.10.511511v1?rss=1</link>
<description><![CDATA[
Safely expanding indications for cellular therapies has been challenging given a lack of highly cancer-specific surface markers. Here, we explore the hypothesis that tumor cells express cancer-specific surface protein conformations, invisible to standard target discovery pipelines evaluating gene or protein expression, that can be identified and immunotherapeutically targeted. We term this strategy, integrating cross-linking mass spectrometry (XL-MS) with glycoprotein surface capture, "structural surfaceomics". As a proof of principle, we apply this technology to acute myeloid leukemia, a hematologic malignancy with dismal outcomes and no known optimal immunotherapy target. We identify the activated conformation of integrin-{beta}2 as a structurally-defined, widely-expressed, AML-specific target. We develop and characterize recombinant antibodies to this protein conformation, and show that chimeric antigen receptor (CAR) T-cells eliminate AML cells and patient-derived xenografts without notable toxicity versus normal hematopoietic cells. Our findings validate an AML conformation-specific target antigen while demonstrating a toolkit for applying these strategies more broadly.
]]></description>
<dc:creator>Mandal, K.</dc:creator>
<dc:creator>Wicaksono, G.</dc:creator>
<dc:creator>Yu, C.</dc:creator>
<dc:creator>Adams, J. J.</dc:creator>
<dc:creator>Hoopmann, M. R.</dc:creator>
<dc:creator>Temple, W. C.</dc:creator>
<dc:creator>Escobar, B. P.</dc:creator>
<dc:creator>Gorelik, M.</dc:creator>
<dc:creator>Ihling, C. H.</dc:creator>
<dc:creator>Nix, M. A.</dc:creator>
<dc:creator>Naik, A.</dc:creator>
<dc:creator>Ramos, E.</dc:creator>
<dc:creator>Kasap, C.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Serrano, J. A. C.</dc:creator>
<dc:creator>Salangsang, F.</dc:creator>
<dc:creator>Phojanakong, P.</dc:creator>
<dc:creator>McMillan, M.</dc:creator>
<dc:creator>Gavallos, V.</dc:creator>
<dc:creator>Leavitt, A. D.</dc:creator>
<dc:creator>Sinz, A.</dc:creator>
<dc:creator>Huang, B. J.</dc:creator>
<dc:creator>Stieglitz, E.</dc:creator>
<dc:creator>Smith, C. C.</dc:creator>
<dc:creator>Moritz, R. L.</dc:creator>
<dc:creator>Sidhu, S. S.</dc:creator>
<dc:creator>Huang, L.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:date>2022-10-11</dc:date>
<dc:identifier>doi:10.1101/2022.10.10.511511</dc:identifier>
<dc:title><![CDATA[Structural surfaceomics reveals an AML-specific conformation of Integrin-β2 as a CAR-T therapy target]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.11.511646v1?rss=1">
<title>
<![CDATA[
A genome-wide CRISPR interference screen using an engineered trafficking biosensor reveals a role for RME-8 in opioid receptor regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.11.511646v1?rss=1</link>
<description><![CDATA[
G protein-coupled receptors (GPCRs) are the largest family of membrane-bound signaling molecules. Activity of these receptors is critically regulated by their trafficking through the endo-lysosomal pathway. Identifying the genes involved in GPCR trafficking is challenging due the complexity of sorting operations and low affinity protein-protein interactions. Here we present a chemical biology fluorescence-based technique to interrogate GPCR trafficking. We show that the engineered enzyme APEX2 is a highly sensitive biosensor for GPCR trafficking to the lysosome, and this trafficking can be monitored through APEX-based activation of fluorogenic substrates such as Amplex UltraRed (AUR). We used this approach to perform a genome-wide CRISPR interference screen focused on the delta type opioid receptor (DOR), a GPCR which modulates anxiety, depression, and pain. The screen identified 492 genes including known- and novel-regulators of DOR expression and trafficking. We demonstrate that one of the novel genes, RME-8, localizes to early endosomes and plays a critical role in regulating DOR trafficking to the lysosome. Together, our data demonstrate that GPCR-APEX2/AUR is a flexible and highly sensitive chemical biology platform for genetic interrogation of receptor trafficking.
]]></description>
<dc:creator>Novy, B.</dc:creator>
<dc:creator>Adoff, H.</dc:creator>
<dc:creator>De Maria, M.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Tsvetanova, N.</dc:creator>
<dc:creator>Von Zastrow, M.</dc:creator>
<dc:creator>Lobingier, B.</dc:creator>
<dc:date>2022-10-11</dc:date>
<dc:identifier>doi:10.1101/2022.10.11.511646</dc:identifier>
<dc:title><![CDATA[A genome-wide CRISPR interference screen using an engineered trafficking biosensor reveals a role for RME-8 in opioid receptor regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.11.511739v1?rss=1">
<title>
<![CDATA[
Therapeutic trial of anle138b in mouse models of genetic prion disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.11.511739v1?rss=1</link>
<description><![CDATA[
Phenotypic screening has yielded small molecule inhibitors of prion replication that are effective in vivo against certain prion strains but not others. Here we sought to test the small molecule anle138b in multiple mouse models of prion disease. In mice inoculated with the RML strain of prions, anle138b doubled survival and durably suppressed astrogliosis measured by live animal bioluminescence imaging. In knock-in mouse models of the D178N and E200K mutations that cause genetic prion disease, however, we were unable to identify a clear, quantifiable disease endpoint against which to measure therapeutic efficacy. Among untreated animals, the mutations did not impact overall survival, and bioluminescence remained low out to >20 months of age. Vacuolization and PrP deposition were observed in some brain regions in a subset of mutant animals, but appeared unable to carry the weight of a primary endpoint in a therapeutic study. We conclude that not all animal models of prion disease are suited to well-powered therapeutic efficacy studies, and care should be taken in choosing the models that will support drug development programs.
]]></description>
<dc:creator>Vallabh, S. M.</dc:creator>
<dc:creator>Zou, D.</dc:creator>
<dc:creator>Pitstick, R.</dc:creator>
<dc:creator>O'Moore, J.</dc:creator>
<dc:creator>Peters, J.</dc:creator>
<dc:creator>Silvius, D.</dc:creator>
<dc:creator>Kriz, J.</dc:creator>
<dc:creator>Jackson, W. S.</dc:creator>
<dc:creator>Carlson, G. A.</dc:creator>
<dc:creator>Minikel, E. V.</dc:creator>
<dc:creator>Cabin, D. E.</dc:creator>
<dc:date>2022-10-12</dc:date>
<dc:identifier>doi:10.1101/2022.10.11.511739</dc:identifier>
<dc:title><![CDATA[Therapeutic trial of anle138b in mouse models of genetic prion disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.12.509557v1?rss=1">
<title>
<![CDATA[
Essential role of the amino-terminal region of Drosha for the Microprocessor function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.12.509557v1?rss=1</link>
<description><![CDATA[
The ribonuclease (RNase) III enzyme Drosha enables processing of microRNA (miRNA) primary transcripts (pri-miRNAs) and control of ribosomal protein (RP) biogenesis by the Microprocessor. The extensively studied carboxyl-terminal half of Drosha is sufficient to crop pri-miRNAs in vitro, but the function of the evolutionarily conserved Drosha amino-terminal region (NTR) is unknown, despite it harboring mutations linked to a hereditary vascular disorder. Here, we provide evidence that, when replacing the endogenous Drosha, a mutant missing the NTR ({Delta}N-Drosha) fails to associate with endogenous pri-miRNAs and to support the expression of all miRNAs, except the miR-183 cluster. Surprisingly, Argonaute2 (Ago2) associates with and cleaves pri-miR-183 in {Delta}N-Drosha cells and in Drosha-depleted cells. {Delta}N-Drosha is also unable to inhibit RP biogenesis upon serum starvation. Thus, Drosha NTR is essential for pri-miRNA processing and RP biogenesis in vivo, and Ago2 can process the miR-183 cluster in the absence of Drosha.
]]></description>
<dc:creator>Hata, A.</dc:creator>
<dc:creator>Prabhakar, A.</dc:creator>
<dc:creator>Jiang, X.</dc:creator>
<dc:creator>Hu, S.</dc:creator>
<dc:creator>Tang, J.</dc:creator>
<dc:creator>Ghatpande, P.</dc:creator>
<dc:creator>Lagna, G.</dc:creator>
<dc:date>2022-10-13</dc:date>
<dc:identifier>doi:10.1101/2022.10.12.509557</dc:identifier>
<dc:title><![CDATA[Essential role of the amino-terminal region of Drosha for the Microprocessor function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.14.512304v1?rss=1">
<title>
<![CDATA[
A unified model for the dynamics of ATP-independent ultrafast contraction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.14.512304v1?rss=1</link>
<description><![CDATA[
In nature, several ciliated protists possess the remarkable ability to execute ultrafast motions using protein assemblies called myonemes, which contract in response to Ca2+ ions. Existing theories, such as actomyosin contractility and macroscopic biomechanical latches, do not adequately describe these systems, necessitating new models to understand their mechanisms. In this study, we image and quantitatively analyze the contractile kinematics observed in two ciliated protists (Vorticella sp and Spirostomum sp), and, based on the mechanochemistry of these organisms, we propose a minimal mathematical model that reproduces our observations as well as those published previously. Analyzing the model reveals three distinct dynamic regimes, differentiated by the rate of chemical driving and the importance of inertia. We characterize their unique scaling behaviors and kinematic signatures. Besides providing insights into Ca2+-powered myoneme contraction in protists, our work may also inform the rational design of ultrafast bioengineered systems such as active synthetic cells.
]]></description>
<dc:creator>Floyd, C.</dc:creator>
<dc:creator>Molines, A. T.</dc:creator>
<dc:creator>Lei, X.</dc:creator>
<dc:creator>Honts, J. E.</dc:creator>
<dc:creator>Chang, F.</dc:creator>
<dc:creator>Elting, M. W.</dc:creator>
<dc:creator>Vaikuntanathan, S.</dc:creator>
<dc:creator>Dinner, A. R.</dc:creator>
<dc:creator>Bhamla, M. S.</dc:creator>
<dc:date>2022-10-16</dc:date>
<dc:identifier>doi:10.1101/2022.10.14.512304</dc:identifier>
<dc:title><![CDATA[A unified model for the dynamics of ATP-independent ultrafast contraction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.14.512312v1?rss=1">
<title>
<![CDATA[
Innate immune cell-intrinsic ketogenesis is dispensable for organismal metabolism and age-related inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.14.512312v1?rss=1</link>
<description><![CDATA[
Aging is accompanied by chronic low-grade inflammation, but the mechanisms that allow this to persist are not well understood. Ketone bodies are alternative fuels produced when glucose is limited and improve indicators of healthspan in aging mouse models. Moreover, the most abundant ketone body, {beta}-hydroxybutyrate (BHB), inhibits the NLRP3 inflammasome in myeloid cells, a key potentiator of age-related inflammation. Given that myeloid cells express ketogenic machinery, we hypothesized this pathway may serve as a metabolic checkpoint of inflammation. To test this hypothesis, we conditionally ablated ketogenesis by disrupting expression of the terminal enzyme required for ketogenesis, 3-Hydroxy-3-Methylglutaryl-CoA Lyase (HMGCL). By deleting HMGCL in the liver, we validated the functional targeting and establish that the liver is the only organ that can produce the life-sustaining quantities of ketone bodies required for survival during fasting or ketogenic diet feeding. Conditional ablation of HMGCL in neutrophils and macrophages had modest effects on body weight and glucose tolerance in agin, but worsened glucose homeostasis in myeloid cell specific Hmgcl deficient mice fed a high-fat diet. Our results suggest that during aging, liver derived circulating ketone bodies might be more important for deactivating NLRP3 inflammasome and controlling organismal metabolism.
]]></description>
<dc:creator>Goldberg, E. L.</dc:creator>
<dc:creator>Letian, A.</dc:creator>
<dc:creator>Dlugos, T.</dc:creator>
<dc:creator>Leveau, C.</dc:creator>
<dc:creator>Dixit, V. D. L.</dc:creator>
<dc:date>2022-10-16</dc:date>
<dc:identifier>doi:10.1101/2022.10.14.512312</dc:identifier>
<dc:title><![CDATA[Innate immune cell-intrinsic ketogenesis is dispensable for organismal metabolism and age-related inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.13.512090v1?rss=1">
<title>
<![CDATA[
Physiological Condition Dependent Changes in Ciliary GPCR Localization in the Brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.13.512090v1?rss=1</link>
<description><![CDATA[
Primary cilia are small immotile cellular appendages which mediate diverse types of singling and are found on most mammalian cell types including throughout the central nervous system. Cilia are known to localize certain G protein-coupled receptors (GPCRs) and are critical for mediating the signaling of these receptors. Several of these neuronal GPCRs have recognized roles in feeding behavior and energy homeostasis. Heterologous cell line and model systems like C. elegans and Chlamydomonas have implicated both dynamic GPCR cilia localization and cilia length and shape changes as key for signaling. However, it is unclear if mammalian ciliary GPCRs utilize similar mechanisms in vivo and under what physiological conditions these processes may occur. Here, we use the ciliary GPCRs, melanin concentrating hormone receptor 1 (MCHR1) and neuropeptide-Y receptor 2 (NPY2R) as model ciliary receptors to determine if dynamic localization to cilia occurs. We tested physiological conditions in which these GPCRs have been implicated such as feeding behavior, obesity, and circadian rhythm. Cilia were imaged using confocal microscopy and analyzed with a computer assisted approach allowing for unbiased and high throughput analysis of cilia. We analyzed GPCR positive cilia, cilia frequency as well as cilia length and receptor occupancy. Interestingly we observed changes in ciliary length, receptor occupancy, and cilia frequency under different conditions, but no consistent theme across GPCRs or brain nuclei was observed. A better understanding of the subcellular localization dynamics of ciliary GPCRs could reveal unrecognized molecular mechanisms regulating behaviors like feeding.

Significance StatementOften, primary cilia localize specific G protein-coupled receptors (GPCRs) for subcellular signaling. Cell lines and model systems have indicated that cilia deploy dynamic GPCR localization and change their shape or length to modulate signaling. We used mice to assess neuronal cilia GPCRs under physiological conditions associated with both the receptors known functions and ciliopathy clinical features like obesity. We show that certain cilia with specific GPCRs appear to dynamically alter their length while others appear relatively stable under these conditions. These results implicate multiple themes across cilia GPCR mediated signaling and indicate that not all cilia modulate GPCR signaling using the same mechanisms. These data will be important for potential pharmacological approaches to target cilia GPCR-mediated signaling.
]]></description>
<dc:creator>Brewer, K. M.</dc:creator>
<dc:creator>Engle, S. E.</dc:creator>
<dc:creator>Bansal, R.</dc:creator>
<dc:creator>Brewer, K. K.</dc:creator>
<dc:creator>Jasso, K. R.</dc:creator>
<dc:creator>McIntyre, J. C.</dc:creator>
<dc:creator>Vaisse, C.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:creator>Berbari, N. F.</dc:creator>
<dc:date>2022-10-17</dc:date>
<dc:identifier>doi:10.1101/2022.10.13.512090</dc:identifier>
<dc:title><![CDATA[Physiological Condition Dependent Changes in Ciliary GPCR Localization in the Brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.13.512183v1?rss=1">
<title>
<![CDATA[
Neurofeedback training can modulate task-relevant memory replay in rats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.13.512183v1?rss=1</link>
<description><![CDATA[
Hippocampal replay - the time-compressed, sequential reactivation of ensembles of neurons related to past experience - is a key neural mechanism of memory consolidation. Replay typically coincides with a characteristic pattern of local field potential activity, the sharp-wave ripple (SWR). Reduced SWR rates are associated with cognitive impairment in multiple models of neurodegenerative disease, suggesting that a clinically viable intervention to promote SWRs and replay would prove beneficial. We therefore developed a neurofeedback paradigm for rat subjects in which SWR detection triggered rapid positive feedback in the context of a memory-dependent task. This training protocol increased the prevalence of task-relevant replay during the targeted neurofeedback period by changing the temporal dynamics of SWR occurrence. This increase was also associated with neural and behavioral forms of compensation after the targeted period. These findings reveal short-timescale regulation of SWR generation and demonstrate that neurofeedback is an effective strategy for modulating hippocampal replay.
]]></description>
<dc:creator>Gillespie, A. K.</dc:creator>
<dc:creator>Astudillo Maya, D. A.</dc:creator>
<dc:creator>Denovellis, E. L.</dc:creator>
<dc:creator>Desse, S.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:date>2022-10-17</dc:date>
<dc:identifier>doi:10.1101/2022.10.13.512183</dc:identifier>
<dc:title><![CDATA[Neurofeedback training can modulate task-relevant memory replay in rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.17.512504v1?rss=1">
<title>
<![CDATA[
Decoding of YAP levels and dynamics by pluripotency factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.17.512504v1?rss=1</link>
<description><![CDATA[
YAP is a transcriptional regulator that controls pluripotency, germ layer specification, and proliferation. Different subsets of YAP target genes are engaged in each physiological setting, but how YAP selectively regulates different effectors in different contexts is not known. Here we use optogenetics to investigate how the levels and dynamics of YAP activation control its pluripotency effectors Oct4 and Nanog. We observe different thresholds for repression of Oct4 and Nanog, enabling differential control of both genes through YAP levels. Pluripotency factors also decode YAP dynamics. Oct4 preferentially responds to oscillatory YAP inputs that mimic endogenous pulsatile YAP dynamics. Using single-cell live imaging of Oct4 transcription and computational-theoretical analysis of transcriptional regulation, we demonstrate that YAP dynamics are decoded by an adaptive change sensor that modulates Oct4 transcription burst frequency. Our results reveal how the levels and timing of YAP activation enable multiplexing of information transmission for key regulators of cellular differentiation and pluripotency.
]]></description>
<dc:creator>Meyer, K.</dc:creator>
<dc:creator>Lammers, N. C.</dc:creator>
<dc:creator>Bugaj, L. J.</dc:creator>
<dc:creator>Garcia, H. H.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:date>2022-10-17</dc:date>
<dc:identifier>doi:10.1101/2022.10.17.512504</dc:identifier>
<dc:title><![CDATA[Decoding of YAP levels and dynamics by pluripotency factors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.14.512299v1?rss=1">
<title>
<![CDATA[
Tracking the online construction of linguistic meaning through negation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.14.512299v1?rss=1</link>
<description><![CDATA[
Combinatoric linguistic operations underpin human language processes, but how meaning is composed and refined in the mind of the reader is not well understood. We address this puzzle by exploiting the ubiquitous function of negation. We track the online effects of negation ("not") and intensifiers ("really") on the representation of scalar adjectives (e.g., "good") in parametrically designed behavioral and neurophysiological (MEG) experiments. The behavioral data show that participants first interpret negated adjectives as affirmative and later modify their interpretation towards, but never exactly as, the opposite meaning. Decoding analyses of neural activity further reveal significant above chance decoding accuracy for negated adjectives within 600 ms from adjective onset, suggesting that negation does not invert the representation of adjectives (i.e., "not bad" represented as "good"); furthermore, decoding accuracy for negated adjectives is found to be significantly lower than that for affirmative adjectives. Overall, these results suggest that negation mitigates rather than inverts the neural representations of adjectives. This putative suppression mechanism of negation is supported by increased synchronization of beta-band neural activity in sensorimotor areas. The analysis of negation provides a steppingstone to understand how the human brain represents changes of meaning over time.
]]></description>
<dc:creator>Zuanazzi, A.</dc:creator>
<dc:creator>Ripolles, P.</dc:creator>
<dc:creator>Lin, W. M.</dc:creator>
<dc:creator>Gwilliams, L.</dc:creator>
<dc:creator>King, J.-R.</dc:creator>
<dc:creator>Poeppel, D.</dc:creator>
<dc:date>2022-10-18</dc:date>
<dc:identifier>doi:10.1101/2022.10.14.512299</dc:identifier>
<dc:title><![CDATA[Tracking the online construction of linguistic meaning through negation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.14.512321v1?rss=1">
<title>
<![CDATA[
CD46 targeted 212Pb alpha particle radioimmunotherapy for prostate cancer treatment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.14.512321v1?rss=1</link>
<description><![CDATA[
We recently identified CD46 as a novel prostate cancer cell surface antigen that shows lineage independent expression in both adenocarcinoma and small cell neuroendocrine subtypes of metastatic castration resistant prostate cancer (mCRPC), discovered an internalizing human monoclonal antibody YS5 that binds to a tumor selective CD46 epitope, and developed a microtubule inhibitor-based antibody drug conjugate that is in a multi-center phase I trial for mCRPC (NCT03575819). Here we report the development of a novel CD46-targeted alpha therapy based on YS5. We conjugated 212Pb, an in vivo generator of alpha-emitting 212Bi and 212Po, to YS5 through the chelator TCMC to create the radioimmunoconjugate, 212Pb-TCMC-YS5. We characterized 212Pb-TCMC-YS5 in vitro and established a safe dose in vivo. We next studied therapeutic efficacy of a single dose of 212Pb-TCMC-YS5 using three prostate cancer small animal models: a subcutaneous mCRPC cell line-derived xenograft (CDX) model (subcu-CDX), an orthotopically grafted mCRPC CDX model (ortho-CDX), and a prostate cancer patient-derived xenograft model (PDX). In all three models, a single dose of 20 Ci 212Pb-TCMC-YS5 was well tolerated and caused potent and sustained inhibition of established tumors, with significant increases of survival in treated animals. A lower dose (10 Ci 212Pb-TCMC-YS5) was also studied on the PDX model, which also showed a significant effect on tumor growth inhibition and prolongation of animal survival. These results demonstrate that 212Pb-TCMC-YS5 has an excellent therapeutic window in preclinical models including PDXs, opening a direct path for clinical translation of this novel CD46-targeted alpha radioimmunotherapy for mCRPC treatment.

SignificanceThis study reports a novel CD46 targeted 212Pb alpha particle radioimmunotherapy, 212Pb-TCMC-YS5, that is well tolerated and shows potent anti-tumor activity (tumor growth inhibition and increase of animal survival) in vivo in three prostate cancer small animal models, i.e., a subcutaneous and an intraprostate orthotopic mCRPC cell line-derived xenograft models, and a prostate cancer patient-derived xenograft model. Given that YS5 is a clinical stage human antibody, this YS5-based 212Pb alpha particle therapy has potential of translation to the clinic for treatment of mCRPC patients.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Huang, T.</dc:creator>
<dc:creator>Hua, J.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Chen, P.</dc:creator>
<dc:creator>Bidlingmaier, S.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Xie, Z.</dc:creator>
<dc:creator>Bidkar, A.</dc:creator>
<dc:creator>Shen, S.</dc:creator>
<dc:creator>Shi, W.</dc:creator>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>Flavell, R. R.</dc:creator>
<dc:creator>Gioeli, D.</dc:creator>
<dc:creator>Dreicer, R.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:date>2022-10-18</dc:date>
<dc:identifier>doi:10.1101/2022.10.14.512321</dc:identifier>
<dc:title><![CDATA[CD46 targeted 212Pb alpha particle radioimmunotherapy for prostate cancer treatment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.17.512583v1?rss=1">
<title>
<![CDATA[
Autism-associated transcriptional regulators target shared loci proximal to brain-expressed genes. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.17.512583v1?rss=1</link>
<description><![CDATA[
Many autism spectrum disorder (ASD)-associated genes act as transcriptional regulators (TRs). ChIP-seq was used to identify the regulatory targets of ARID1B, BCL11A, FOXP1, TBR1, and TCF7L2, ASD-associated TRs in the developing human and mouse cortex. These TRs shared substantial overlap in the binding sites, especially within open chromatin. The overlap within a promoter region, 1-2,000bp upstream of transcription start site, was highly predictive of brain expressed genes. This signature was observed at 96 out of 102 ASD-associated genes. In vitro CRISPRi against ARID1B and TBR1 delineated downstream convergent biology in mouse cortical cultures. After eight days, NeuN+ and CALB+ cells were decreased, GFAP+ cells were increased, and transcriptomic signatures correlated with the postmortem brain samples from individuals with ASD. We suggest functional convergence across five ASD-associated TRs leads to shared neurodevelopmental outcomes of haploinsufficient disruption.
]]></description>
<dc:creator>Fazel Darbandi, S.</dc:creator>
<dc:creator>An, J. Y.</dc:creator>
<dc:creator>Lim, K.</dc:creator>
<dc:creator>Page, N. F.</dc:creator>
<dc:creator>Liang, L.</dc:creator>
<dc:creator>Ypsilanti, A.</dc:creator>
<dc:creator>Markenscoff-Papadimitriou, E.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Nord, A. S.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Rubenstein, J. L. R.</dc:creator>
<dc:date>2022-10-18</dc:date>
<dc:identifier>doi:10.1101/2022.10.17.512583</dc:identifier>
<dc:title><![CDATA[Autism-associated transcriptional regulators target shared loci proximal to brain-expressed genes.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.17.512631v1?rss=1">
<title>
<![CDATA[
7SK methylation Promotes Transcriptional Activity upon Growth Factor Stimulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.17.512631v1?rss=1</link>
<description><![CDATA[
A fundamental facet of cell signaling is the conversion of extracellular signals into adaptive transcriptional responses. The role of RNA modifications in this process is poorly understood. The small nuclear RNA 7SK prevents transcription elongation by sequestering the complex CDK9/CCNT1 (P-TEFb). We discovered that METTL3 methylates 7SK. The m6A methylation of 7SK in turn promotes its binding to heterogeneous nuclear ribonucleoproteins (HNRNPs), with consequent release of the HEXIM1/P-TEFb complex - leading to the induction of growth factor-stimulated transcriptional responses. The methylation of 7SK relies on the activation of METTL3 via phosphorylation downstream of growth factors-signaling pathways such as the epidermal growth factor (EGF). Our findings establish a novel function for the m6A modification in converting growth-factor signaling events to a transcriptional elongation regulatory response via an RNA-methylation-dependent switch.

One-Sentence Summarym6A methylation of the non-coding RNA 7SK promotes transcriptional activity upon growth factor stimulation.
]]></description>
<dc:creator>Alarcon, C. R.</dc:creator>
<dc:creator>Perez-Pepe, M.</dc:creator>
<dc:creator>Desotell, A. W.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Han, B.</dc:creator>
<dc:creator>Lin, Q.</dc:creator>
<dc:creator>Klein, D. E.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2022-10-18</dc:date>
<dc:identifier>doi:10.1101/2022.10.17.512631</dc:identifier>
<dc:title><![CDATA[7SK methylation Promotes Transcriptional Activity upon Growth Factor Stimulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.18.512708v1?rss=1">
<title>
<![CDATA[
Impact of SARS-CoV-2 ORF6 and its variant polymorphisms on host responses and viral pathogenesis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.18.512708v1?rss=1</link>
<description><![CDATA[
We and others have previously shown that the SARS-CoV-2 accessory protein ORF6 is a powerful antagonist of the interferon (IFN) signaling pathway by directly interacting with Nup98-Rae1 at the nuclear pore complex (NPC) and disrupting bidirectional nucleo-cytoplasmic trafficking. In this study, we further assessed the role of ORF6 during infection using recombinant SARS-CoV-2 viruses carrying either a deletion or a well characterized M58R loss-of-function mutation in ORF6. We show that ORF6 plays a key role in the antagonism of IFN signaling and in viral pathogenesis by interfering with karyopherin(importin)-mediated nuclear import during SARS-CoV-2 infection both in vitro, and in the Syrian golden hamster model in vivo. In addition, we found that ORF6-Nup98 interaction also contributes to inhibition of cellular mRNA export during SARS-CoV-2 infection. As a result, ORF6 expression significantly remodels the host cell proteome upon infection. Importantly, we also unravel a previously unrecognized function of ORF6 in the modulation of viral protein expression, which is independent of its function at the nuclear pore. Lastly, we characterized the ORF6 D61L mutation that recently emerged in Omicron BA.2 and BA.4 and demonstrated that it is able to disrupt ORF6 protein functions at the NPC and to impair SARS-CoV-2 innate immune evasion strategies. Importantly, the now more abundant Omicron BA.5 lacks this loss-of-function polymorphism in ORF6. Altogether, our findings not only further highlight the key role of ORF6 in the antagonism of the antiviral innate immune response, but also emphasize the importance of studying the role of non-spike mutations to better understand the mechanisms governing differential pathogenicity and immune evasion strategies of SARS-CoV-2 and its evolving variants.

ONE SENTENCE SUMMARYSARS-CoV-2 ORF6 subverts bidirectional nucleo-cytoplasmic trafficking to inhibit host gene expression and contribute to viral pathogenesis.
]]></description>
<dc:creator>Kehrer, T.</dc:creator>
<dc:creator>Cupic, A.</dc:creator>
<dc:creator>Ye, C.</dc:creator>
<dc:creator>Yildiz, S.</dc:creator>
<dc:creator>Bouhhadou, M.</dc:creator>
<dc:creator>Crossland, N. A.</dc:creator>
<dc:creator>Barrall, E.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Tseng, A.</dc:creator>
<dc:creator>Cagatay, T.</dc:creator>
<dc:creator>Rathnasinghe, R.</dc:creator>
<dc:creator>Flores, D.</dc:creator>
<dc:creator>Jangra, S.</dc:creator>
<dc:creator>Alam, F.</dc:creator>
<dc:creator>Mena, N.</dc:creator>
<dc:creator>Aslam, S.</dc:creator>
<dc:creator>Saqi, A.</dc:creator>
<dc:creator>Marin, A.</dc:creator>
<dc:creator>Rutkowska, M.</dc:creator>
<dc:creator>Ummadi, M. R.</dc:creator>
<dc:creator>Pisanelli, G.</dc:creator>
<dc:creator>Richardson, R. B.</dc:creator>
<dc:creator>Veit, E. C.</dc:creator>
<dc:creator>Fabius, J. M. A.</dc:creator>
<dc:creator>Soucheray, M.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Evans, M. J.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Gonzalez-Reiche, A. S.</dc:creator>
<dc:creator>Sordillo, E. M.</dc:creator>
<dc:creator>van Bakel, H.</dc:creator>
<dc:creator>Simon, V.</dc:creator>
<dc:creator>Zuliani-Alvarez, L.</dc:creator>
<dc:creator>Fontoura, B. M. A.</dc:creator>
<dc:creator>Rosenberg, B. R.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Martinez-Sobrido, L.</dc:creator>
<dc:creator>Garcia-Sastre, A.</dc:creator>
<dc:creator>Miorin, L.</dc:creator>
<dc:date>2022-10-19</dc:date>
<dc:identifier>doi:10.1101/2022.10.18.512708</dc:identifier>
<dc:title><![CDATA[Impact of SARS-CoV-2 ORF6 and its variant polymorphisms on host responses and viral pathogenesis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.19.512772v1?rss=1">
<title>
<![CDATA[
Long-term, super-resolution HIDE imaging of the inner mitochondrial membrane in live cells with a cell-permeant lipid probe 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.19.512772v1?rss=1</link>
<description><![CDATA[
The densely packed inner mitochondrial membrane (IMM) is vital for bioenergy generation and its dynamics control mitochondrial health and cellular homeostasis. IMM structure is complex, however, and imaging its dynamics with high temporal and spatial resolution is complicated by the photosensitivity of IMM-resident enzymes. Here we describe the cell-permeant, lipid-like acridine orange derivative MAO-N3 and use it to assemble high-density, environmentally sensitive (HIDE) probes that selectively label and image the IMM in live cells. MAO-N3 pairs with multiple SPAAC-reactive fluorophores to support HIDE imaging via confocal, Structured Illumination, Single Molecule Localization, and Stimulated Emission Depletion microscopy, all with significantly improved resistance against photobleaching. The HIDE probes generated using MAO-N3 require no genetic manipulations, are non-toxic in model cell lines and primary cardiomyocytes, even under conditions that amplify the effects of mitochondrial toxins, and visualize the IMM for up to 12.5 hours with unprecedented spatial and temporal resolution.
]]></description>
<dc:creator>Zheng, S.</dc:creator>
<dc:creator>Dadina, N.</dc:creator>
<dc:creator>Mozumdar, D.</dc:creator>
<dc:creator>Lesiak, L.</dc:creator>
<dc:creator>Martinez, K.</dc:creator>
<dc:creator>Miller, E.</dc:creator>
<dc:creator>Schepartz, A.</dc:creator>
<dc:date>2022-10-19</dc:date>
<dc:identifier>doi:10.1101/2022.10.19.512772</dc:identifier>
<dc:title><![CDATA[Long-term, super-resolution HIDE imaging of the inner mitochondrial membrane in live cells with a cell-permeant lipid probe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.19.512927v1?rss=1">
<title>
<![CDATA[
Global landscape of the host response to SARS-CoV-2 variants reveals viral evolutionary trajectories 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.19.512927v1?rss=1</link>
<description><![CDATA[
A series of SARS-CoV-2 variants of concern (VOCs) have evolved in humans during the COVID-19 pandemic--Alpha, Beta, Gamma, Delta, and Omicron. Here, we used global proteomic and genomic analyses during infection to understand the molecular responses driving VOC evolution. We discovered VOC-specific differences in viral RNA and protein expression levels, including for N, Orf6, and Orf9b, and pinpointed several viral mutations responsible. An analysis of the host response to VOC infection and comprehensive interrogation of altered virus-host protein-protein interactions revealed conserved and divergent regulation of biological pathways. For example, regulation of host translation was highly conserved, consistent with suppression of VOC replication in mice using the translation inhibitor plitidepsin. Conversely, modulation of the host inflammatory response was most divergent, where we found Alpha and Beta, but not Omicron BA.1, antagonized interferon stimulated genes (ISGs), a phenotype that correlated with differing levels of Orf6. Additionally, Delta more strongly upregulated proinflammatory genes compared to other VOCs. Systematic comparison of Omicron subvariants revealed BA.5 to have evolved enhanced ISG and proinflammatory gene suppression that similarly correlated with Orf6 expression, effects not seen in BA.4 due to a mutation that disrupts the Orf6-nuclear pore interaction. Our findings describe how VOCs have evolved to fine-tune viral protein expression and protein-protein interactions to evade both innate and adaptive immune responses, offering a likely explanation for increased transmission in humans.

One sentence summarySystematic proteomic and genomic analyses of SARS-CoV-2 variants of concern reveal how variant-specific mutations alter viral gene expression, virus-host protein complexes, and the host response to infection with applications to therapy and future pandemic preparedness.
]]></description>
<dc:creator>Bouhaddou, M.</dc:creator>
<dc:creator>Reuschl, A.-K.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Thorne, L. G.</dc:creator>
<dc:creator>Ummadi, M. R.</dc:creator>
<dc:creator>Ye, C.</dc:creator>
<dc:creator>Ramirez, R. R.</dc:creator>
<dc:creator>Pelin, A.</dc:creator>
<dc:creator>Batra, J.</dc:creator>
<dc:creator>Jang, G. M.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Moen, J. M.</dc:creator>
<dc:creator>Richards, A. L.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Harjai, B.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Rojc, A.</dc:creator>
<dc:creator>Ragazzini, R.</dc:creator>
<dc:creator>Whelan, M. V. X.</dc:creator>
<dc:creator>Furnon, W.</dc:creator>
<dc:creator>De Lorenzo, G.</dc:creator>
<dc:creator>Cowton, V.</dc:creator>
<dc:creator>Syed, A. M.</dc:creator>
<dc:creator>Ciling, A.</dc:creator>
<dc:creator>Deutsch, N.</dc:creator>
<dc:creator>Pirak, D.</dc:creator>
<dc:creator>Dowgier, G.</dc:creator>
<dc:creator>Mesner, D.</dc:creator>
<dc:creator>Turner, J. L.</dc:creator>
<dc:creator>McGovern, B. L.</dc:creator>
<dc:creator>Rodriguez, M. L.</dc:creator>
<dc:creator>Leiva-Rebollo, R.</dc:creator>
<dc:creator>Dunham, A. S.</dc:creator>
<dc:creator>Zhong, X.</dc:creator>
<dc:creator>Eckhardt, M.</dc:creator>
<dc:creator>Fossati, A.</dc:creator>
<dc:creator>Liotta, N.</dc:creator>
<dc:creator>Kehrer, T.</dc:creator>
<dc:creator>Cupic, A.</dc:creator>
<dc:creator>Rutkowska, M.</dc:creator>
<dc:creator>Mena, N.</dc:creator>
<dc:creator>Aslam, S.</dc:creator>
<dc:creator>Hoffert, A.</dc:creator>
<dc:creator>Foussard, H.</dc:creator>
<dc:creator>Pham, J.</dc:creator>
<dc:creator>Ly</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.19.512927</dc:identifier>
<dc:title><![CDATA[Global landscape of the host response to SARS-CoV-2 variants reveals viral evolutionary trajectories]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.19.512943v1?rss=1">
<title>
<![CDATA[
The IRE1α-XBP1 signaling axis promotes glycolytic reprogramming in response to inflammatory stimuli 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.19.512943v1?rss=1</link>
<description><![CDATA[
Immune cells must be able to adjust their metabolic programs to effectively carry out their effector functions. Here, we show that the ER stress sensor IRE1 and its downstream transcription factor XBP1 enhance the upregulation of glycolysis in classically activated macrophages (CAM). The IRE1-XBP1 signaling axis supports this glycolytic switch in macrophages when activated by LPS stimulation or infection with the intracellular bacterial pathogen Brucella abortus. Importantly, these different inflammatory stimuli have distinct mechanisms of IRE1 activation; while TLR4 supports glycolysis under both conditions, TLR4 is required for activation of IRE1 in response to LPS treatment but not B. abortus infection. Though IRE1 and XBP1 are necessary for maximal induction of glycolysis in CAM, activation of this pathway is not sufficient to increase the glycolytic rate of macrophages, indicating that the cellular context in which this pathway is activated ultimately dictates the cells metabolic response and that IRE1 activation may be a way to fine-tune metabolic reprogramming.

IMPORTANCEThe immune system must be able to tailor its response to different types of pathogens in order to eliminate them and protect the host. When confronted with bacterial pathogens, macrophages, frontline defenders in the immune system, switch to a glycolysis-driven metabolism to carry out their antibacterial functions. Here, we show that IRE1, a sensor of ER stress, and its downstream transcription factor XBP1 support glycolysis in macrophages during infection with Brucella abortus or challenge with Salmonella LPS. Interestingly, these stimuli activate IRE1 by independent mechanisms. While the IRE1-XBP1 signaling axis promotes the glycolytic switch, activation of this pathway is not sufficient to increase glycolysis in macrophages. This study furthers our understanding of the pathways that drive macrophage immunometabolism and highlights a new role for IRE1 and XBP1 in innate immunity.
]]></description>
<dc:creator>English, B. C.</dc:creator>
<dc:creator>Savage, H. P.</dc:creator>
<dc:creator>Mahan, S. P.</dc:creator>
<dc:creator>Diaz-Ochoa, V. E.</dc:creator>
<dc:creator>Young, B. M.</dc:creator>
<dc:creator>Abuaita, B. H.</dc:creator>
<dc:creator>Sule, G. J.</dc:creator>
<dc:creator>Knight, J. S.</dc:creator>
<dc:creator>O'Riordan, M. X.</dc:creator>
<dc:creator>Baumler, A. J.</dc:creator>
<dc:creator>Tsolis, R. M.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.19.512943</dc:identifier>
<dc:title><![CDATA[The IRE1α-XBP1 signaling axis promotes glycolytic reprogramming in response to inflammatory stimuli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.20.512880v1?rss=1">
<title>
<![CDATA[
Species-specific roles for the MAFA and MAFB transcription factors in regulatingislet β cell identity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.20.512880v1?rss=1</link>
<description><![CDATA[
Type 2 diabetes (T2D) is associated with compromised identity of insulin-producing pancreatic islet beta ({beta}) cells, characterized by inappropriate production of other islet cell-enriched hormones. Here we examined how hormone misexpression was influenced by the MAFA and MAFB transcription factors, closely related proteins that maintain islet cell function. Mice specifically lacking MafA in {beta} cells demonstrated broad, population-wide changes in hormone gene expression with an overall gene signature closely resembling islet gastrin (Gast)-positive cells generated under conditions of chronic hyperglycemia and obesity. A human {beta} cell line deficient in MAFB, but not one lacking MAFA, also produced a gastrin (GAST)-positive gene expression pattern. In addition, GAST was detected in human T2D {beta} cells with low levels of MAFB. Moreover, evidence is provided that human MAFB can directly repress GAST gene transcription. These results support a novel, species-specific role for MafA and MAFB in maintaining adult mouse and human {beta} cell identity, respectively, by repressing expression of Gast/GAST and other non-{beta} cell hormones.
]]></description>
<dc:creator>Cha, J.</dc:creator>
<dc:creator>Tong, X.</dc:creator>
<dc:creator>Walker, E. M.</dc:creator>
<dc:creator>Dahan, T.</dc:creator>
<dc:creator>Cochrane, V.</dc:creator>
<dc:creator>Ashe, S.</dc:creator>
<dc:creator>Russell, R.</dc:creator>
<dc:creator>Osipovich, A. B.</dc:creator>
<dc:creator>Mawla, A. M.</dc:creator>
<dc:creator>Guo, M.</dc:creator>
<dc:creator>Liu, J.-h.</dc:creator>
<dc:creator>Huising, M. O.</dc:creator>
<dc:creator>Magnuson, M. A.</dc:creator>
<dc:creator>Hebrok, M.</dc:creator>
<dc:creator>Dor, Y.</dc:creator>
<dc:creator>Stein, R.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.20.512880</dc:identifier>
<dc:title><![CDATA[Species-specific roles for the MAFA and MAFB transcription factors in regulatingislet β cell identity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.20.512885v1?rss=1">
<title>
<![CDATA[
A reversible SRC-relayed COX2-inflammatory program drives therapeutic resistance in BRAF(V600E) colorectal tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.20.512885v1?rss=1</link>
<description><![CDATA[
BRAFV600E mutation confers a poor prognosis in metastatic colorectal cancer (CRC) despite combinatorial targeted therapies based on the latest understanding of signaling circuitry. To identify parallel resistance mechanisms induced by BRAF/MEK/EGFR co-targeting, we used a high throughput kinase activity mapping platform. We found that SRC kinases are systematically activated in BRAFV600E CRC following targeted inhibition of BRAF {+/-} EGFR, and that coordinated targeting of SRC with BRAF {+/-} EGFR increases efficacy in vitro and in vivo. SRC drives resistance to BRAF {+/-} anti-EGFR therapy independently of ERK signaling by inducing transcriptional reprogramming via beta-catenin (CTNNB1). The EGFR-independent compensatory activation of SRC kinases is mediated by an autocrine prostaglandin E2-loop that can be blocked with cyclooxygenase-2 (COX2) inhibitors. Co-targeting of COX2 with BRAF+EGFR promotes durable suppression of tumor growth in patient-derived tumor xenograft (PDX) models. COX2 inhibition represents a novel drug-repurposing strategy to overcome therapeutic resistance in BRAFV600E CRC.
]]></description>
<dc:creator>Ruiz-Saenz, A.</dc:creator>
<dc:creator>Atreya, C. E.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Pan, B.</dc:creator>
<dc:creator>Dreyer, C. A.</dc:creator>
<dc:creator>Brunen, D.</dc:creator>
<dc:creator>Prahallad, A.</dc:creator>
<dc:creator>Munoz, D. P.</dc:creator>
<dc:creator>Ramms, D. J.</dc:creator>
<dc:creator>Burghi, V.</dc:creator>
<dc:creator>Spassov, D. S.</dc:creator>
<dc:creator>Fewings, E.</dc:creator>
<dc:creator>Hwang, Y. C.</dc:creator>
<dc:creator>Cowdrey, C.</dc:creator>
<dc:creator>Moelders, C.</dc:creator>
<dc:creator>Schwarzer, C.</dc:creator>
<dc:creator>Wolf, D. M.</dc:creator>
<dc:creator>Hann, B.</dc:creator>
<dc:creator>VandenBerg, S. R.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:creator>Moasser, M. M.</dc:creator>
<dc:creator>Bernards, R.</dc:creator>
<dc:creator>Gutkind, J. S.</dc:creator>
<dc:creator>van't Veer, L. J.</dc:creator>
<dc:creator>Coppe, J.-P.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.20.512885</dc:identifier>
<dc:title><![CDATA[A reversible SRC-relayed COX2-inflammatory program drives therapeutic resistance in BRAF(V600E) colorectal tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.24.513565v1?rss=1">
<title>
<![CDATA[
Rubella virus tropism and single cell responses in human primary tissue and microglia-containing organoids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.24.513565v1?rss=1</link>
<description><![CDATA[
Rubella virus is an important human pathogen that can cause neurologic deficits in a developing fetus when contracted during pregnancy. Despite successful vaccination programs in the Americas and many developed countries, rubella remains endemic in many regions worldwide and outbreaks occur wherever population immunity is insufficient. Intense interest since rubella virus was first isolated in 1962 has advanced our understanding of clinical outcomes after infection disrupts key processes of fetal neurodevelopment. Yet it is still largely unknown which cell types in the developing brain are targeted. We show that in human brain slices, rubella virus predominantly infects microglia. This infection occurs in a heterogeneous population but not in a highly microglia-enriched monoculture in the absence of other cell types. By using an organoid-microglia model, we further demonstrate that rubella virus infection leads to a profound interferon response in non-microglial cells, including neurons and neural progenitor cells, and this response is attenuated by the presence of microglia.
]]></description>
<dc:creator>Popova, G.</dc:creator>
<dc:creator>Retallack, H.</dc:creator>
<dc:creator>Kim, C. N.</dc:creator>
<dc:creator>Shin, D.</dc:creator>
<dc:creator>Wang, A.</dc:creator>
<dc:creator>DeRisi, J. J.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513565</dc:identifier>
<dc:title><![CDATA[Rubella virus tropism and single cell responses in human primary tissue and microglia-containing organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.24.513434v1?rss=1">
<title>
<![CDATA[
A general approach to discover conformational epitope-directed binders for selective inhibition of protein proteolytic activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.24.513434v1?rss=1</link>
<description><![CDATA[
Directing antibodies to a particular epitope among many possible on a target protein is a significant challenge. Here we present a simple and general method for epitope-directed selection (EDS) using a differential phage selection strategy. This involves engineering the protein of interest (POI) with the epitope of interest (EOI) mutated using a systematic bioinformatics algorithm to guide the local design of an EOI decoy variant. Using several alternating rounds of negative selection with the EOI decoy variant followed by positive selection on the wild-type (WT) POI, we were able to identify highly specific and potent antibodies to five different EOI antigens that bind and functionally block known sites of proteolysis. Among these we developed highly specific antibodies that target the proteolytic site on the CUB domain containing protein 1 (CDCP1) to prevent its proteolysis allowing us to study the cellular maturation of this event that triggers malignancy. We generated antibodies that recognize the junction between the pro and catalytic domains for four different matrix metalloproteases (MMPs), such as MMP1, MMP3, and MMP9, that selectively block activation of each of these enzymes and impairs cell migration. We targeted a proteolytic epitope on the cell surface receptor, EPH Receptor A2, that is known to transform it from a tumor suppressor to an oncoprotein. We believe the EDS method greatly facilitates the generation antibodies to specific EOIs on a wide range of proteins and enzymes for broad therapeutic and diagnostic applications.

SignificanceWe have developed a highly efficient platform to facilitate the directed selection in vitro of antibodies to a wide range of functional epitopes on proteins. This method uses a bioinformatic program to guide mutations in the local site of interest to create a decoy antigen that can effectively remove antibodies not binding the site of interest by negative selection, followed by positive selection with the WT antigen to identify antibodies to the epitope of interest. We demonstrate the generality and versatility of this method by successfully producing functional antibodies to block specific proteolytically sensitive epitopes on five different proteins including enzymes important in cancer. The epitope-directed selection (EDS) approach greatly facilitates the identification of binders to specific sites of interest on proteins to probe function and as potential immunotherapeutics.
]]></description>
<dc:creator>Zhou, J.</dc:creator>
<dc:date>2022-10-25</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513434</dc:identifier>
<dc:title><![CDATA[A general approach to discover conformational epitope-directed binders for selective inhibition of protein proteolytic activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.24.513541v1?rss=1">
<title>
<![CDATA[
A cross-species proteomic map of synapse development reveals neoteny during human postsynaptic density maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.24.513541v1?rss=1</link>
<description><![CDATA[
The molecular mechanisms and evolutionary changes accompanying synapse development are still poorly understood. Here, we generated a cross-species proteomic map of synapse development in the human, macaque, and mouse neocortex. By tracking the changes of >1,000 postsynaptic density (PSD) proteins from midgestation to young adulthood, we found that PSD maturation in humans separates into three major phases that are dominated by distinct pathways. Cross-species comparisons reveal that the human PSD matures about two to three times slower than other species and contains higher levels of Rho guanine nucleotide exchange factors (RhoGEFs) in the perinatal period. Enhancement of the RhoGEF signaling in human neurons delays the morphological maturation of dendritic spines and the functional maturation of synapses, potentially contributing to the neotenic traits of human brain development. In addition, PSD proteins can be divided into four modules that exert stage- and cell type-specific functions, possibly explaining their differential associations with cognitive functions and diseases. Together, our proteomic map of synapse development provides a blueprint for studying the molecular basis and evolutionary changes of synapse maturation.
]]></description>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Pang, K.</dc:creator>
<dc:creator>Zhou, L.</dc:creator>
<dc:creator>Cebrian-Silla, A.</dc:creator>
<dc:creator>Gonzalez-Granero, S.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Bi, Q.</dc:creator>
<dc:creator>White, M. L.</dc:creator>
<dc:creator>Ho, B.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Perez, Y.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Winkler, E. A.</dc:creator>
<dc:creator>Paredes, M. F.</dc:creator>
<dc:creator>Kovner, R.</dc:creator>
<dc:creator>Sestan, N.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:creator>Liu, P.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Piao, X.</dc:creator>
<dc:creator>Garcia-Verdugo, J. M.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Kriegstein, A. R.</dc:creator>
<dc:date>2022-10-25</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513541</dc:identifier>
<dc:title><![CDATA[A cross-species proteomic map of synapse development reveals neoteny during human postsynaptic density maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.24.513555v1?rss=1">
<title>
<![CDATA[
Single-cell analysis of prenatal and postnatal human cortical development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.24.513555v1?rss=1</link>
<description><![CDATA[
We analyze more than 700,000 single-nucleus RNA-seq profiles from 106 donors during prenatal and postnatal developmental stages and identify lineage-specific programs that underlie the development of specific subtypes of excitatory cortical neurons, interneurons, glial cell types and brain vasculature. By leveraging single-nucleus chromatin accessibility data, we delineate enhancer-gene regulatory networks and transcription factors that control commitment of specific cortical lineages. By intersecting our results with genetic risk factors for human brain diseases, we identify the cortical cell types and lineages most vulnerable to genetic insults of different brain disorders, especially autism. We find that lineage-specific gene expression programs upregulated in female cells are especially enriched for the genetic risk factors of autism. Our study captures the molecular progression of cortical lineages across human development.

One Sentence SummarySingle-cell transcriptomic atlas of human cortical development identifies lineage and sex-specific programs and their implication in brain disorders.
]]></description>
<dc:creator>Velmeshev, D.</dc:creator>
<dc:creator>Perez, Y.</dc:creator>
<dc:creator>Yan, Z.</dc:creator>
<dc:creator>Valencia, J. E.</dc:creator>
<dc:creator>Castaneda-Castellanos, D. R.</dc:creator>
<dc:creator>Schirmer, L.</dc:creator>
<dc:creator>Mayer, S.</dc:creator>
<dc:creator>Wick, B.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Paredes, M.</dc:creator>
<dc:creator>Huang, E. J. J.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:date>2022-10-25</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513555</dc:identifier>
<dc:title><![CDATA[Single-cell analysis of prenatal and postnatal human cortical development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.24.513566v1?rss=1">
<title>
<![CDATA[
Granulin Loss of Function in Human Mature Brain Organoids Implicates Astrocytes in TDP-43 Pathology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.24.513566v1?rss=1</link>
<description><![CDATA[
Loss of function (LoF) of Tar-binding protein 43 (TDP-43) and mislocalization, together with TDP-43-positive and hyperphosphorylated inclusions, are found in postmortem tissue of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) patients, including those carrying LoF variants in the progranulin gene (GRN). Modelling TDP-43 pathology has been challenging in vivo and in vitro. We present a 3D-induced pluripotent stem cell (iPSC)-derived paradigm - mature brain organoids (mbOrg) - composed of cortical-like-astrocytes (iA) and neurons (iN). When devoid of GRN, mbOrgs spontaneously recapitulate TDP-43 mislocalization, hyperphosphorylation and LoF phenotypes. Mixing-and-matching genotypes in mbOrgs showed that GRN-/- iA are drivers for TDP-43 pathology. Finally, we rescued TDP-43 LoF by adding exogenous progranulin, demonstrating a link between TDP-43 LoF and progranulin expression. In conclusion, we present an iPSC-derived platform that shows striking features of human TDP-43 proteinopathy and provides a tool for mechanistic modelling of TDP-43 pathology and patient-tailored therapeutic screening for FTD and ALS.

HighlightsO_LIGRN-/- iPSC-derived 3D paradigm (mbOrg) composed of mature cortical-like astrocytes and neurons recapitulates features of TDP-43 proteinopathy
C_LIO_LIGRN-/- cortical-like astrocytes are necessary and sufficient for the development of the TDP-43 loss of function phenotype in mbOrg.
C_LIO_LIA TDP-43 phenotype can be rescued in neurons by treating neuron and astrocyte co-cultures with progranulin full length protein.
C_LI

eTOC blurbIn this article, de Majo and colleagues present a novel 3D iPSC-derived model to study neurodegenerative disorders such as ALS and FTD. When devoid of GRN expression, these cultures present features of ALS and FTD associated pathology hardly ever observed in vitro. These phenotypes are shown to be primarily driven by diseased astrocytes and can be rescued by progranulin supplementation.
]]></description>
<dc:creator>de Majo, M.</dc:creator>
<dc:creator>Koontz, M.</dc:creator>
<dc:creator>Marsan, E.</dc:creator>
<dc:creator>Salinas, N.</dc:creator>
<dc:creator>Ramsey, A.</dc:creator>
<dc:creator>Kuo, Y.-M.</dc:creator>
<dc:creator>Seo, K.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Draeger, N. M.</dc:creator>
<dc:creator>Leng, K.</dc:creator>
<dc:creator>Kurnellas, M.</dc:creator>
<dc:creator>Miyaoka, Y.</dc:creator>
<dc:creator>Klim, J. R.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Ward, M. E.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Ullian, E. M.</dc:creator>
<dc:date>2022-10-25</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513566</dc:identifier>
<dc:title><![CDATA[Granulin Loss of Function in Human Mature Brain Organoids Implicates Astrocytes in TDP-43 Pathology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.24.513607v1?rss=1">
<title>
<![CDATA[
Loss of MLL3/4 decouples enhancer H3K4 monomethylation, H3K27 acetylation, and gene activation during ESC differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.24.513607v1?rss=1</link>
<description><![CDATA[
Enhancers are essential in defining cell fates through the control of cell type specific gene expression. Enhancer activation is a multi-step process involving chromatin remodelers and histone modifiers including the monomethylation of H3K4 (H3K4me1) by MLL3 (KMT2C) and MLL4 (KMT2D). MLL3/4 are thought to be critical for enhancer activation and cognate gene expression including through the recruitment of acetyltransferases for H3K27. Here we test this model by evaluating the impact of MLL3/4 loss on chromatin and transcription during early embryonic stem cell differentiation. We find that MLL3/4 activity is required at most if not all sites that gain or lose H3K4me1 but is largely dispensable at sites that remain stably methylated during this transition. This requirement extends to H3K27 acetylation (H3K27ac) at most transitional sites. However, many sites gain H3K27ac independent of MLL3/4 or H3K4me1 including enhancers regulating key factors in early differentiation. Furthermore, despite the failure to gain active histone marks at thousands of enhancers, transcriptional activation of nearby genes is largely unaffected, thus uncoupling the regulation of these chromatin events from transcriptional changes during this transition. These data challenge current models of enhancer activation and imply distinct mechanisms between stable and dynamically changing enhancers. Collectively, our study highlights gaps in knowledge about the steps and epistatic relationships of enzymes necessary for enhancer activation and cognate gene transcription.
]]></description>
<dc:creator>Blelloch, R.</dc:creator>
<dc:creator>Boileau, R. M.</dc:creator>
<dc:creator>Chen, K. X.</dc:creator>
<dc:date>2022-10-25</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513607</dc:identifier>
<dc:title><![CDATA[Loss of MLL3/4 decouples enhancer H3K4 monomethylation, H3K27 acetylation, and gene activation during ESC differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.25.513779v1?rss=1">
<title>
<![CDATA[
The global anaerobic metabolism regulator fnr is necessary for the degradation of food dyes and drugs by Escherichia coli 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.25.513779v1?rss=1</link>
<description><![CDATA[
The microbiome is an underappreciated contributor to intestinal drug metabolism with broad implications for drug efficacy and toxicity. While considerable progress has been made towards identifying the gut bacterial genes and enzymes involved, the role of environmental factors in shaping their activity remains poorly understood. Here, we focus on the gut bacterial reduction of azo bonds (R-N=N-R), found in diverse chemicals in both food and drugs. Surprisingly, the canonical azoR gene in Escherichia coli was dispensable for azo bond reduction. Instead, azo reductase activity was controlled by the fumarate and nitrate reduction (fnr) regulator, consistent with a requirement for the anoxic conditions found within the gastrointestinal tract. Paired transcriptomic and proteomic analysis of the fnr regulon revealed that in addition to altering the expression of multiple reductases, FNR is necessary for the metabolism of L-Cysteine to hydrogen sulfide, enabling the degradation of azo bonds. Taken together, these results show how gut bacteria sense and respond to their intestinal environment to enable the metabolism of chemical motifs found in both dietary and pharmaceutical compounds.

IMPORTANCEThis work has broad relevance due to the ubiquity of dyes containing azo bonds in food and drugs. We report that azo dyes can be degraded by human gut bacteria through both enzymatic and non-enzymatic mechanisms, even from a single gut bacterial species. Furthermore, we revealed that environmental factors, oxygen and cysteine, control the ability of E. coli to degrade azo dyes due to their impacts on bacterial transcription and metabolism. These results open up new opportunities to manipulate the azoreductase activity of the gut microbiome through the manipulation of host diet, suggest that azoreductase potential may be altered in patients suffering from gastrointestinal disease, and highlight the importance of studying bacterial enzymes for drug metabolism in their natural cellular and ecological context.
]]></description>
<dc:creator>Pieper, L. M.</dc:creator>
<dc:creator>Spanogiannopoulos, P.</dc:creator>
<dc:creator>Volk, R.</dc:creator>
<dc:creator>Miller, C.</dc:creator>
<dc:creator>Wright, A.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:date>2022-10-25</dc:date>
<dc:identifier>doi:10.1101/2022.10.25.513779</dc:identifier>
<dc:title><![CDATA[The global anaerobic metabolism regulator fnr is necessary for the degradation of food dyes and drugs by Escherichia coli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.25.513743v1?rss=1">
<title>
<![CDATA[
Dissecting the effects of GTPase and kinase domain mutations on LRRK2 endosomal localization and activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.25.513743v1?rss=1</link>
<description><![CDATA[
Parkinsons disease-causing LRRK2 mutations lead to varying degrees of Rab GTPase hyperphosphorylation. Puzzlingly, LRRK2 GTPase-inactivating mutations--which do not affect intrinsic kinase activity--lead to higher levels of cellular Rab phosphorylation than kinase-activating mutations. Here, we investigated whether mutation-dependent differences in LRRK2 cellular localization could explain this discrepancy. We discovered that blocking endosomal maturation leads to the rapid formation of mutant LRRK2+ endosomes on which LRRK2 phosphorylates substrate Rabs. LRRK2+ endosomes are maintained through positive feedback, which mutually reinforces membrane localization of LRRK2 and phosphorylated Rab substrates. Furthermore, across a panel of mutants, cells expressing GTPase-inactivating mutants formed strikingly more LRRK2+ endosomes than cells expressing kinase-activating mutants, resulting in higher total cellular levels of phosphorylated Rabs. Our study suggests that an increased probability of LRRK2 GTPase-inactivating mutants to be retained on intracellular membranes over the kinase-activating mutants leads to higher substrate phosphorylation.
]]></description>
<dc:creator>Rinaldi, C.</dc:creator>
<dc:creator>Waters, C. S.</dc:creator>
<dc:creator>Kumbier, K.</dc:creator>
<dc:creator>Rao, L.</dc:creator>
<dc:creator>Nichols, R. J.</dc:creator>
<dc:creator>Jacobson, M. P.</dc:creator>
<dc:creator>Wu, L. F.</dc:creator>
<dc:creator>Altschuler, S. J.</dc:creator>
<dc:date>2022-10-27</dc:date>
<dc:identifier>doi:10.1101/2022.10.25.513743</dc:identifier>
<dc:title><![CDATA[Dissecting the effects of GTPase and kinase domain mutations on LRRK2 endosomal localization and activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.26.513935v1?rss=1">
<title>
<![CDATA[
A deficiency screen of the 3rd chromosome for dominant modifiers of the Drosophila ER integral membrane protein, Jagunal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.26.513935v1?rss=1</link>
<description><![CDATA[
The mechanism surrounding chromosome inheritance during cell division has been well documented, however, organelle inheritance during mitosis is less understood. Recently, the Endoplasmic Reticulum (ER) has been shown to reorganize during mitosis, dividing asymmetrically in proneuronal cells prior to cell fate selection, indicating a programmed mechanism of inheritance. ER asymmetric partitioning in proneural cells relies on the highly conserved ER integral membrane protein, Jagunal (Jagn). Knockdown of Jagn in the compound Drosophila eye displays a pleotropic rough eye phenotype in 48% of the progeny. To identify genes involved in Jagn dependent ER partitioning pathway, we performed a dominant modifier screen of the 3rd chromosome for enhancers and suppressors of this Jagn RNAi-induced rough eye phenotype. We screened through 181 deficiency lines covering the 3L and 3R chromosomes and identified 12 suppressors and 10 enhancers of the Jagn RNAi phenotype. Based on the functions of the genes covered by the deficiencies, we identified genes that displayed a suppression or enhancement of the Jagn RNAi phenotype. These include Division Abnormally Delayed (Dally), an heparan sulfate proteoglycan, the {gamma}-secretase subunit Presenilin, and the ER resident protein Sec63. Based on our understanding of the function of these targets, there is a connection between Jagn and the Notch signaling pathway. Further studies will elucidate the role of Jagn and identified interactors within the mechanisms of ER partitioning during mitosis.
]]></description>
<dc:creator>Ascencio, G.</dc:creator>
<dc:creator>de Cruz, M. A.</dc:creator>
<dc:creator>Abuel, J.</dc:creator>
<dc:creator>Alvarado, S.</dc:creator>
<dc:creator>Arriaga, Y.</dc:creator>
<dc:creator>Conrad, E. A.</dc:creator>
<dc:creator>Castro, A.</dc:creator>
<dc:creator>Eichelberger, K.</dc:creator>
<dc:creator>Galvan, L.</dc:creator>
<dc:creator>Gundy, G.</dc:creator>
<dc:creator>Inojoza, J. A.</dc:creator>
<dc:creator>Jimenez, A. D.</dc:creator>
<dc:creator>Lu, N. T.</dc:creator>
<dc:creator>Lugar, C.</dc:creator>
<dc:creator>Marania, R.</dc:creator>
<dc:creator>Mendsaikhan, T.</dc:creator>
<dc:creator>Ortega, J.</dc:creator>
<dc:creator>Nand, N.</dc:creator>
<dc:creator>Rodrigues, N. S.</dc:creator>
<dc:creator>Shabazz, K.</dc:creator>
<dc:creator>Tam, C.</dc:creator>
<dc:creator>Valenciano, E.</dc:creator>
<dc:creator>Hayzelden, C.</dc:creator>
<dc:creator>Eritano, A. S.</dc:creator>
<dc:creator>Riggs, B.</dc:creator>
<dc:date>2022-10-28</dc:date>
<dc:identifier>doi:10.1101/2022.10.26.513935</dc:identifier>
<dc:title><![CDATA[A deficiency screen of the 3rd chromosome for dominant modifiers of the Drosophila ER integral membrane protein, Jagunal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.27.513961v1?rss=1">
<title>
<![CDATA[
Membrane-active peptides escape drug-resistance in cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.27.513961v1?rss=1</link>
<description><![CDATA[
Acquired drug-resistance is a recurring problem in cancer treatment, and this is particularly true for patients with metastatic melanoma that carry a BRAF V600E mutation. In the current study, we explored the use of membrane-active peptides as an alternative therapeutic modality to target drug-resistant melanoma cells. We produced slow-cycling and drug-resistant melanoma cells using dabrafenib, a small molecule drug that targets tumor cells with BRAF V600E mutation, and characterised their lipidome and proteome to investigate the role of membrane lipids in acquired drug-resistance. Despite some changes in the lipid composition, tested anti-melanoma membrane-active cyclic peptides (cTI and cGm) killed melanoma cells that are sensitive, tolerant, or resistant to dabrafenib. Importantly, melanoma cells did not develop resistance to cTI or cGm, nor changed their lipid composition with long-term peptide treatment. Therefore, these peptides are well suited as templates to design therapeutic leads to target drug-resistant metastatic melanoma cells and/or as co-treatment with small molecule drugs.
]]></description>
<dc:creator>Benfield, A. H.</dc:creator>
<dc:creator>Vernen, F.</dc:creator>
<dc:creator>Young, R. S. E.</dc:creator>
<dc:creator>Bufi, F. N.</dc:creator>
<dc:creator>Hammerlindl, H.</dc:creator>
<dc:creator>Craik, D. J.</dc:creator>
<dc:creator>Schaider, H.</dc:creator>
<dc:creator>Lawrence, N.</dc:creator>
<dc:creator>Blanksby, S. J.</dc:creator>
<dc:creator>Henriques, S. T.</dc:creator>
<dc:date>2022-10-28</dc:date>
<dc:identifier>doi:10.1101/2022.10.27.513961</dc:identifier>
<dc:title><![CDATA[Membrane-active peptides escape drug-resistance in cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.28.514160v1?rss=1">
<title>
<![CDATA[
Linking Choroid Plexus Enlargement with Plasma Analyte and Structural Phenotypes in Clinical High Risk for Psychosis: A Multisite Neuroimaging Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.28.514160v1?rss=1</link>
<description><![CDATA[
BackgroundChoroid plexus (ChP) enlargement exists in first-episode and chronic psychosis, but whether enlargement occurs before psychosis onset is unknown. This study investigated whether ChP volume is enlarged in individuals with clinical high-risk (CHR) for psychosis and whether these changes are related to clinical, neuroanatomical, and plasma analytes.

MethodsClinical and neuroimaging data from the North American Prodrome Longitudinal Study 2 (NAPLS2) was used for analysis. 509 participants (169 controls, 340 CHR) were recruited. Conversion status was determined after 2-years of follow-up, with 36 psychosis converters. The lateral ventricle ChP was manually segmented from baseline scans. A subsample of 31 controls and 53 CHR had plasma analyte and neuroimaging data.

ResultsCompared to controls, CHR (d=0.23, p=0.017) and non-converters (d=0.22, p=0.03) demonstrated higher ChP volumes, but not in converters. In CHR, greater ChP volume correlated with lower cortical (r=-0.22, p<0.001), subcortical gray matter (r=-0.21, p<0.001), and total white matter volume (r=-0.28,p<0.001), as well as larger lateral ventricle volume (r=0.63,p<0.001). Greater ChP volume correlated with makers functionally associated with the lateral ventricle ChP in CHR [CCL1 (r=-0.30, p=0.035), ICAM1 (r=0.33, p=0.02)], converters [IL1{beta} (r=0.66, p=0.004)], and non-converters [BMP6 (r=-0.96, p<0.001), CALB1 (r=-0.98, p<0.001), ICAM1 (r=0.80, p=0.003), SELE (r=0.59, p=0.026), SHBG (r=0.99, p<0.001), TNFRSF10C (r=0.78, p=0.001)].

ConclusionsCHR and non-converters demonstrated significantly larger ChP volumes compared to controls. Enlarged ChP was associated with neuroanatomical alterations and analyte markers functionally associated with the ChP. These findings suggest that the ChP may be a key explanatory biomarker in CHR for psychosis.
]]></description>
<dc:creator>Bannai, D.</dc:creator>
<dc:creator>Reuter, M.</dc:creator>
<dc:creator>Hegde, R.</dc:creator>
<dc:creator>Hoang, D.</dc:creator>
<dc:creator>Adhan, I.</dc:creator>
<dc:creator>Gandu, S.</dc:creator>
<dc:creator>Pong, S.</dc:creator>
<dc:creator>Zeng, A.</dc:creator>
<dc:creator>Raymond, N.</dc:creator>
<dc:creator>Zeng, V.</dc:creator>
<dc:creator>Chung, Y.</dc:creator>
<dc:creator>He, G.</dc:creator>
<dc:creator>Sun, D.</dc:creator>
<dc:creator>van Erp, T. G. M.</dc:creator>
<dc:creator>Addington, J.</dc:creator>
<dc:creator>Bearden, C. E.</dc:creator>
<dc:creator>Cadenhead, K.</dc:creator>
<dc:creator>Cornblatt, B.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>McGlashan, T.</dc:creator>
<dc:creator>Jeffries, C.</dc:creator>
<dc:creator>Stone, W.</dc:creator>
<dc:creator>Tsuang, M.</dc:creator>
<dc:creator>Walker, E.</dc:creator>
<dc:creator>Woods, S. W.</dc:creator>
<dc:creator>Cannon, T. D.</dc:creator>
<dc:creator>Perkins, D.</dc:creator>
<dc:creator>Keshavan, M.</dc:creator>
<dc:creator>Lizano, P.</dc:creator>
<dc:date>2022-10-30</dc:date>
<dc:identifier>doi:10.1101/2022.10.28.514160</dc:identifier>
<dc:title><![CDATA[Linking Choroid Plexus Enlargement with Plasma Analyte and Structural Phenotypes in Clinical High Risk for Psychosis: A Multisite Neuroimaging Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.28.514304v1?rss=1">
<title>
<![CDATA[
Single-cell RNA-seq reveals alterations in peripheral CX3CR1 and nonclassical monocytes in familial tauopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.28.514304v1?rss=1</link>
<description><![CDATA[
BackgroundEmerging evidence from mouse models is beginning to elucidate the brains immune response to tau pathology, but little is known about the nature of this response in humans. In addition, it remains unclear to what extent tau pathology and the local inflammatory response within the brain influence the broader immune system.

MethodsTo address these questions, we performed single-cell RNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) from carriers of pathogenic variants in MAPT, the gene encoding tau.

ResultsAnalysis of [~]181,000 individual PBMC transcriptomes from MAPT pathogenic variant carriers (n = 8) and healthy non-carrier controls (n = 8) demonstrated striking differential expression in monocytes and natural killer (NK) cells. We observed a marked reduction in the expression of CX3CR1 - the gene encoding the fractalkine receptor that is known to modulate tau pathology in mouse models - in monocytes and NK cells. We also observed a significant reduction in the abundance of nonclassical monocytes and dysregulated expression of nonclassical monocyte marker genes, including FCGR3A. Finally, we identified reductions in TMEM176A and TMEM176B, genes thought to be involved in the inflammatory response in human microglia. We confirmed differential expression of select biologically relevant genes dysregulated in our scRNA-seq data using droplet digital PCR as an orthogonal technique for quantitative validation.

ConclusionsOur results suggest that human peripheral immune cell expression and abundance are modulated by tau-associated pathophysiologic changes. CX3CR1 and nonclassical monocytes in particular will be a focus of future work exploring the role of these peripheral signals in additional tau-associated neurodegenerative diseases.
]]></description>
<dc:creator>Sirkis, D. W.</dc:creator>
<dc:creator>Warly Solsberg, C.</dc:creator>
<dc:creator>Johnson, T. P.</dc:creator>
<dc:creator>Bonham, L. W.</dc:creator>
<dc:creator>Sturm, V. E.</dc:creator>
<dc:creator>Lee, S. E.</dc:creator>
<dc:creator>Rankin, K. P.</dc:creator>
<dc:creator>Rosen, H. J.</dc:creator>
<dc:creator>Boxer, A. L.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Geier, E. G.</dc:creator>
<dc:creator>Yokoyama, J. S.</dc:creator>
<dc:date>2022-10-30</dc:date>
<dc:identifier>doi:10.1101/2022.10.28.514304</dc:identifier>
<dc:title><![CDATA[Single-cell RNA-seq reveals alterations in peripheral CX3CR1 and nonclassical monocytes in familial tauopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.30.514414v1?rss=1">
<title>
<![CDATA[
Robust expression of transgenes in Drosophila melanogaster 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.30.514414v1?rss=1</link>
<description><![CDATA[
Drosophila is a classic experimental system used in fundamental and biopharmaceutical research. Manipulating gene expression in the larvae and adult flies can facilitate basic and translational studies. Here we report a method for robust transgene overexpression in Drosophila melanogaster. This approach is compatible with the UAS/Gal4 gene expression system. The improved expression involves a gene expression cassette that prevents transgenic mRNA transcript degradation by the nonsense-mediated RNA decay pathway. The protection is mediated by CrPV and DCV IRES sequences that apparently encode cryptic polyadenylation sites and NMD inhibitors. Stabilization of the transgene mRNA results in a >40-fold increase in its levels.
]]></description>
<dc:creator>Lidsky, P. V.</dc:creator>
<dc:creator>Dmitriev, S. E.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:date>2022-10-31</dc:date>
<dc:identifier>doi:10.1101/2022.10.30.514414</dc:identifier>
<dc:title><![CDATA[Robust expression of transgenes in Drosophila melanogaster]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.31.514594v1?rss=1">
<title>
<![CDATA[
Excessive Firing of Dyskinesia-Associated Striatal Direct Pathway Neurons is Gated By Dopamine and Excitatory Synaptic Input 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.31.514594v1?rss=1</link>
<description><![CDATA[
The striatum integrates dopaminergic and glutamatergic inputs to select preferred versus alternative actions, but the precise mechanisms remain unclear. One way to study action selection is when it breaks down. Here, we explored the cellular and synaptic mechanisms of levodopa-induced dyskinesia (LID), a complication of Parkinsons disease therapy characterized by involuntary movements. We used an activity-dependent tool (FosTRAP) in conjunction with a mouse model of LID to investigate functionally distinct subsets of striatal direct pathway medium spiny neurons (dMSNs). In vivo, levodopa differentially activates dyskinesia-associated (TRAPed) dMSNs compared to other dMSNs. This activation is likely to be driven by two cellular mechanisms we identified through ex vivo electrophysiology: higher sensitivity to dopamine and stronger excitatory input from the motor cortex and thalamus. Together, these findings suggest how intrinsic and synaptic properties of heterogeneous dMSN subpopulations integrate to support action selection.
]]></description>
<dc:creator>Ryan, M. B.</dc:creator>
<dc:creator>Girasole, A. E.</dc:creator>
<dc:creator>McGregor, M. M.</dc:creator>
<dc:creator>Brakaj, R.</dc:creator>
<dc:creator>Paletzki, R. F.</dc:creator>
<dc:creator>Gerfen, C. R.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2022-11-01</dc:date>
<dc:identifier>doi:10.1101/2022.10.31.514594</dc:identifier>
<dc:title><![CDATA[Excessive Firing of Dyskinesia-Associated Striatal Direct Pathway Neurons is Gated By Dopamine and Excitatory Synaptic Input]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.01.514771v1?rss=1">
<title>
<![CDATA[
A Semi-Supervised Ensemble Approach to Rank Potential Causal Variants and Their Target Genes in Microglia for Alzheimer's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.01.514771v1?rss=1</link>
<description><![CDATA[
Alzheimers disease (AD) is the leading cause of death among individuals over 65. Despite many AD genetic variants detected by large genome-wide association studies (GWAS), a limited number of causal genes have been confirmed. Conventional machine learning techniques integrate functional annotation data and GWAS signals to assign variants functional relevance probabilities. Yet, a large proportion of genetic variation lies in the non-coding genome, where unsupervised and semi-supervised techniques have demonstrated greater advantage. Furthermore, cell-type specific approaches are needed to better understand disease etiology. Studying AD from a microglia-specific lens is more likely to reveal causal variants involved in immune pathways. Therefore, in this study, we developed S-BEAM: a semi-supervised ensemble approach using microglia-specific data to prioritize non-coding variants and their target genes that play roles in immune-related AD mechanisms. We designed a transductive positive-unlabeled and negative-unlabeled learning model that employs a bagging technique to learn from unlabeled variants, generating multiple predicted probabilities of variant risk. Using a combined homogeneous-heterogeneous ensemble framework, we aggregated the predictions. We applied our model to AD variant data, identifying 11 risk variants acting in well-known AD genes, such as TSPAN14, INPP5D, and MS4A2. These results validated our models performance and demonstrated a need to study these genes in the context of microglial pathways. We also proposed further experimental study for 37 potential causal variants associated with less-known genes. Our work has utility in predicting AD relevant genes and variants functioning in microglia and can be generalized for application to other complex diseases or cell types.
]]></description>
<dc:creator>Khaire, A.</dc:creator>
<dc:creator>Wen, J.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:date>2022-11-03</dc:date>
<dc:identifier>doi:10.1101/2022.11.01.514771</dc:identifier>
<dc:title><![CDATA[A Semi-Supervised Ensemble Approach to Rank Potential Causal Variants and Their Target Genes in Microglia for Alzheimer's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.02.514751v1?rss=1">
<title>
<![CDATA[
Room-temperature crystallography reveals altered binding of small-molecule fragments to PTP1B 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.02.514751v1?rss=1</link>
<description><![CDATA[
Much of our current understanding of how small-molecule ligands interact with proteins stems from X-ray crystal structures determined at cryogenic (cryo) temperature. For proteins alone, room-temperature (RT) crystallography can reveal previously hidden, biologically relevant alternate conformations. However, less is understood about how RT crystallography may impact the conformational landscapes of protein-ligand complexes. Previously we showed that small-molecule fragments cluster in putative allosteric sites using a cryo crystallographic screen of the therapeutic target PTP1B (Keedy*, Hill*, 2018). Here we have performed two RT crystallographic screens of PTP1B using many of the same fragments, representing the largest RT crystallographic screens of a diverse library of ligands to date, and enabling a direct interrogation of the effect of data collection temperature on protein-ligand interactions. We show that at RT, fewer ligands bind, and often more weakly -- but with a variety of temperature-dependent differences, including unique binding poses, changes in solvation, new binding sites, and distinct protein allosteric conformational responses. Overall, this work suggests that the vast body of existing cryogenic-temperature protein-ligand structures may provide an incomplete picture, and highlights the potential of RT crystallography to help complete this picture by revealing distinct conformational modes of protein-ligand systems. Our results may inspire future use of RT crystallography to interrogate the roles of protein-ligand conformational ensembles in biological function.
]]></description>
<dc:creator>Mehlman, T. S.</dc:creator>
<dc:creator>Biel, J. T.</dc:creator>
<dc:creator>Azeem, S. M.</dc:creator>
<dc:creator>Nelson, E. R.</dc:creator>
<dc:creator>Hossain, S.</dc:creator>
<dc:creator>Dunnett, L. E.</dc:creator>
<dc:creator>Paterson, N. G.</dc:creator>
<dc:creator>Douangamath, A.</dc:creator>
<dc:creator>Talon, R.</dc:creator>
<dc:creator>Axford, D.</dc:creator>
<dc:creator>Orins, H.</dc:creator>
<dc:creator>von Delft, F.</dc:creator>
<dc:creator>Keedy, D. A.</dc:creator>
<dc:date>2022-11-03</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514751</dc:identifier>
<dc:title><![CDATA[Room-temperature crystallography reveals altered binding of small-molecule fragments to PTP1B]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.02.514868v1?rss=1">
<title>
<![CDATA[
Eos promotes TH2 differentiation by propagating the IL-2/STAT5 signaling pathway. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.02.514868v1?rss=1</link>
<description><![CDATA[
The Ikaros zinc finger transcription factor Eos has been commonly implicated in regulatory T cells to promote their immunosuppressive functions. Paradoxically, a new role is emerging for Eos in promoting pro-inflammatory responses of conventional CD4+ T cells in the dysregulated setting of autoimmunity. Even so, the precise role of Eos in regulating the differentiation and function of healthy effector CD4+ T cell subsets remains unclear. Here, we find that Eos is a positive regulator of CD4+ T helper 2 (TH2) cells--effector T cells implicated in the induction of allergic asthma. Using murine in vitro TH2 cells and an in vivo house dust mite asthma model, we found that Eos-deficient T cells had reduced expression of key TH2 transcription factors, effector cytokines, and differentiation receptors. Mechanistically, among various TH2-polarizing pathways, the IL-2/STAT5 axis and its downstream TH2 gene targets emerged as one of the most significantly downregulated networks in Eos deficiency. Using in vitro TH2 cells and overexpression of Eos zinc-finger-domain mutants, we discovered that Eos forms a novel complex with and supports the tyrosine-phosphorylated signaling activity of STAT5. Overall, these data define a novel regulatory mechanism whereby Eos promotes IL-2/STAT5 activity to facilitate TH2 differentiation.
]]></description>
<dc:creator>Tuazon, J. A.</dc:creator>
<dc:creator>Read, K. A.</dc:creator>
<dc:creator>Sreekumar, B. K.</dc:creator>
<dc:creator>Yaeger, M. J.</dc:creator>
<dc:creator>Varikuti, S.</dc:creator>
<dc:creator>Jones, D. M.</dc:creator>
<dc:creator>Warren, R. T.</dc:creator>
<dc:creator>Powell, M. D.</dc:creator>
<dc:creator>Rasheed, M. N.</dc:creator>
<dc:creator>Duncan, E. G.</dc:creator>
<dc:creator>Childs, L. M.</dc:creator>
<dc:creator>Gowdy, K. M.</dc:creator>
<dc:creator>Oestreich, K. J.</dc:creator>
<dc:date>2022-11-03</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514868</dc:identifier>
<dc:title><![CDATA[Eos promotes TH2 differentiation by propagating the IL-2/STAT5 signaling pathway.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.02.514888v1?rss=1">
<title>
<![CDATA[
Ras-dependent RAF-MAPK hyperactivation by pathogenic RIT1 is a therapeutic target in Noonan syndrome-associated cardiac hypertrophy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.02.514888v1?rss=1</link>
<description><![CDATA[
RIT1 belongs to the family of Ras guanosine triphosphatases (GTPases) that regulate many aspects of signal transduction and are drivers of cancer and congenital disorders. RIT1 gain-of-function mutations are found in lung cancer, leukemia, and in the germline of Noonan syndrome individuals with an increased prevalence of cardiac hypertrophy and other congenital heart defects. Pathogenic RIT1 proteins evade proteasomal degradation and promote MEK/ERK mitogen-activated protein kinase (MAPK) hyperactivation, yet the mechanism remains poorly understood. Here we show that RAF kinases are putative mutant RIT1 effectors necessary for MAPK activation and characterize RIT1 association with plasma membrane lipids and interaction with RAF kinases. We identify critical residues present in the RIT1 hypervariable region that facilitate interaction with negatively charged membrane lipids and show that these are necessary for association with RAF kinases. Although mutant RIT1 binds to RAF kinases directly, it fails to activate RAF-MAPK signaling in the absence of classical Ras proteins. Consistent with aberrant RAF/MEK/ERK activation as a driver of disease, we show that MEK inhibition alleviates cardiac hypertrophy in a mouse model of RIT1-mutant Noonan syndrome. These data shed light on pathogenic RIT1 function and identify avenues for therapeutic intervention.

One Sentence SummaryElectrostatic plasma membrane association facilitates RIT1-mediated Ras-dependent RAF kinase activation to promote pathogenic MAPK signaling.
]]></description>
<dc:creator>Cuevas-Navarro, A.</dc:creator>
<dc:creator>Wagner, M.</dc:creator>
<dc:creator>Van, R.</dc:creator>
<dc:creator>Swain, M.</dc:creator>
<dc:creator>Allison, M. R.</dc:creator>
<dc:creator>Cheng, A.</dc:creator>
<dc:creator>Messing, S.</dc:creator>
<dc:creator>Simanshu, D. K.</dc:creator>
<dc:creator>Sale, M. J.</dc:creator>
<dc:creator>McCormick, F.</dc:creator>
<dc:creator>Stephen, A. G.</dc:creator>
<dc:creator>Castel, P.</dc:creator>
<dc:date>2022-11-03</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514888</dc:identifier>
<dc:title><![CDATA[Ras-dependent RAF-MAPK hyperactivation by pathogenic RIT1 is a therapeutic target in Noonan syndrome-associated cardiac hypertrophy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.02.514938v1?rss=1">
<title>
<![CDATA[
Parabolic avalanche scaling in the synchronization of cortical cell assemblies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.02.514938v1?rss=1</link>
<description><![CDATA[
Neurons in cortex synchronize their spiking in response to local and distant inputs. These synchronized assemblies are fundamental to cortex function, yet basic dynamical aspects about their size and duration are largely unknown. Using 2-photon imaging of neurons in superficial cortex of awake mice, we show that synchronized assemblies organize as scale-invariant avalanches that quadratically grow with duration. This quadratic expansion was found only for correlated neurons and required temporal coarse graining to compensate for spatial subsampling when network dynamics are critical, as demonstrated in simulations. The corresponding time course of an inverted parabola with exponent of {chi} = 2 described avalanches of up to 5 s duration and maximized temporal complexity in the ongoing activity of prefrontal and somatosensory cortex and in visual responses of primary visual cortex. Our results identify a scale-invariant order in the synchronization of highly diverse cortical cell assemblies in the form of parabolic avalanches.

Significance StatementThe synchronization of cell assemblies is fundamental to many brain theories. Here we show such synchronization to grow according to an inverted parabola that maximizes temporal complexity. This quadratic scaling found for cell assemblies of highly diverse size and duration is in line with prediction for neuronal avalanches and the cortex being in a critical state.
]]></description>
<dc:creator>Capek, E.</dc:creator>
<dc:creator>Lins-Ribeiro, T.</dc:creator>
<dc:creator>Srinivasan, K.</dc:creator>
<dc:creator>Miller, S. R.</dc:creator>
<dc:creator>Victor, M.</dc:creator>
<dc:creator>Geist, E.</dc:creator>
<dc:creator>Vakili, A.</dc:creator>
<dc:creator>Pajevic, S.</dc:creator>
<dc:creator>Chialvo, D. R.</dc:creator>
<dc:creator>Plenz, D.</dc:creator>
<dc:date>2022-11-03</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514938</dc:identifier>
<dc:title><![CDATA[Parabolic avalanche scaling in the synchronization of cortical cell assemblies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.02.514809v1?rss=1">
<title>
<![CDATA[
Multi-model order spatially constrained ICA reveals highly replicable group differences and consistent predictive results from fMRI data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.02.514809v1?rss=1</link>
<description><![CDATA[
Brain functional networks identified from resting fMRI data have the potential to reveal biomarkers for brain disorders, but studies of complex mental illnesses such as schizophrenia (SZ) often yield mixed results across replication studies. This is likely due in part to the complexity of the disorder, the short data acquisition time, and the limited ability of the approaches for brain imaging data mining. Therefore, the use of analytic approaches which can both capture individual variability while offering comparability across analyses is highly preferred. Fully blind data-driven approaches such as independent component analysis (ICA) are hard to compare across studies, and approaches that use fixed atlas-based regions can have limited sensitivity to individual sensitivity. By contrast, spatially constrained ICA (scICA) provides a hybrid, fully automated solution that can incorporate spatial network priors while also adapting to new subjects. However, scICA has thus far only been used with a single spatial scale. In this work, we present an approach using scICA to extract subject-specific intrinsic connectivity networks (ICNs) from fMRI data at multiple spatial scales (ICA model orders), which also enables us to study interactions across spatial scales. We evaluate this approach using a large N (N>1,600) study of schizophrenia divided into separate validation and replication sets. A multi-scale ICN template was estimated and labeled, then used as input into spatially constrained ICA which was computed on an individual subject level. We then performed a subsequent analysis of multiscale functional network connectivity (msFNC) to evaluate the patient data, including group differences and classification. Results showed highly consistent group differences in msFNC in regions including cerebellum, thalamus, and motor/auditory networks. Importantly, multiple msFNC pairs linking different spatial scales were implicated. We also used the msFNC features as input to a classification model in cross-validated hold-out data and also in an independent test data. Visualization of predictive features was performed by evaluating their feature weights. Finally, we evaluated the relationship of the identified patterns to positive symptoms and found consistent results across datasets. The results verified the robustness of our framework in evaluating brain functional connectivity of schizophrenia at multiple spatial scales, implicated consistent and replicable brain networks, and highlighted a promising approach for leveraging resting fMRI data for brain biomarker development.
]]></description>
<dc:creator>Meng, X.</dc:creator>
<dc:creator>Iraji, A.</dc:creator>
<dc:creator>Fu, Z.</dc:creator>
<dc:creator>Kochunov, P.</dc:creator>
<dc:creator>Belger, A.</dc:creator>
<dc:creator>Ford, J.</dc:creator>
<dc:creator>McEwen, S.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>Mueller, B. A.</dc:creator>
<dc:creator>Pearlson, G. D.</dc:creator>
<dc:creator>Potkin, S. G.</dc:creator>
<dc:creator>Preda, A.</dc:creator>
<dc:creator>Turner, J.</dc:creator>
<dc:creator>Erp, T. G. M. v.</dc:creator>
<dc:creator>Sui, J.</dc:creator>
<dc:creator>Calhoun, V.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514809</dc:identifier>
<dc:title><![CDATA[Multi-model order spatially constrained ICA reveals highly replicable group differences and consistent predictive results from fMRI data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.02.514913v1?rss=1">
<title>
<![CDATA[
Cellpanelr: identify predictive biomarkers from cell line panel response data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.02.514913v1?rss=1</link>
<description><![CDATA[
SummaryCellpanelr is an open-source R package and web application for analyzing user-generated cell panel screens using DepMap data sets. Cellpanelr can be used to identify mutation and expression biomarkers of cell line response, increasing the value and accessibility of cell panel experiments such as relative sensitivities to cancer drugs.

Availability and implementationHosted web application is available from shinyapps.io (https://dwassarman.shinyapps.io/cellpanelr). Source code and installation instructions are available from GitHub (https://github.com/dwassarman/cellpanelr).

Contactdwassar@gmail.com
]]></description>
<dc:creator>Wassarman, D. R.</dc:creator>
<dc:creator>Wu, T.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514913</dc:identifier>
<dc:title><![CDATA[Cellpanelr: identify predictive biomarkers from cell line panel response data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.03.515075v1?rss=1">
<title>
<![CDATA[
Astrocyte-like CEPsh glia responds to the stressful environments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.03.515075v1?rss=1</link>
<description><![CDATA[
While astrocytes are known to be important for development and nourishment of the nervous system, the field is just beginning to explore how astrocytes respond to environmental stimuli. Using Caenorhabditis elegans and their astrocyte-equivalent, CEPsh, we asked whether astrocyte-like glia respond to odor exposure. We found one day-old adult C. elegans decrease hlh-17 promoter (CEPsh glia marker) mediated fluorescent expression when trained with an innately attractive odor butanone. Moreover, the olfactory training paradigm itself affects phlh-17 expression, but in a different way. This suggests astrocyte-like CEPsh glia can integrate environmental information to respond to changes in the environment, which enhances survival.
]]></description>
<dc:creator>Chien, E.</dc:creator>
<dc:creator>Garcia, A. J.</dc:creator>
<dc:creator>CHANDRA, R.</dc:creator>
<dc:creator>L'Etoile, N.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.03.515075</dc:identifier>
<dc:title><![CDATA[Astrocyte-like CEPsh glia responds to the stressful environments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.06.515311v1?rss=1">
<title>
<![CDATA[
Integrative single-cell characterization of hypothalamus sex-differential and obesity-associated genes and regulatory elements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.06.515311v1?rss=1</link>
<description><![CDATA[
Over 500 noncoding genomic loci are associated with obesity. The majority of these loci reside near genes that are expressed in the hypothalamus in specific neuronal subpopulations that regulate food intake, hindering the ability to identify and functionally characterize them. Here, we carried out integrative single-cell analysis (RNA/ATAC-seq) on both mouse and human male and female hypothalamus to characterize genes and regulatory elements in specific cell subpopulations. Utilizing both transcriptome and regulome data, we identify over 30 different neuronal and non-neuronal cell subpopulations and a shared core of transcription factors that regulate cell cluster-specific genes between mice and humans. We characterize several sex-specific differentially expressed genes and the regulatory elements that control them in specific cell subpopulations. Overlapping cell-specific scATAC peaks with obesity-associated GWAS variants, identifies potential obesity-associated regulatory elements. Using reporter assays and CRISPR editing, we show that many of these sequences, including the top obesity-associated loci (FTO and MC4R), are functional enhancers whose activity is altered due to the obesity-associated variant and regulate known obesity genes. Combined, our work provides a catalog of genes and regulatory elements in hypothalamus cell subpopulations and uses obesity to showcase how integrative single-cell sequencing can identify functional variants associated with hypothalamus-related phenotypes.
]]></description>
<dc:creator>Nguyen, H. P.</dc:creator>
<dc:creator>Chan, C. S.</dc:creator>
<dc:creator>Cintron, D. L.</dc:creator>
<dc:creator>Sheng, R.</dc:creator>
<dc:creator>Harshman, L.</dc:creator>
<dc:creator>Nobuhara, M.</dc:creator>
<dc:creator>Ushiki, A.</dc:creator>
<dc:creator>Biellak, C.</dc:creator>
<dc:creator>An, K.</dc:creator>
<dc:creator>Gordon, G. M.</dc:creator>
<dc:creator>Mifsud, F.</dc:creator>
<dc:creator>Blake, A.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Hemberg, M.</dc:creator>
<dc:creator>Vaisse, C.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2022-11-06</dc:date>
<dc:identifier>doi:10.1101/2022.11.06.515311</dc:identifier>
<dc:title><![CDATA[Integrative single-cell characterization of hypothalamus sex-differential and obesity-associated genes and regulatory elements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.06.515335v1?rss=1">
<title>
<![CDATA[
Cholinergic Interneurons Drive Motivation by Promoting Dopamine Release in the Nucleus Accumbens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.06.515335v1?rss=1</link>
<description><![CDATA[
Motivation to work for potential rewards is critically dependent on dopamine (DA) in the nucleus accumbens (NAc). DA release from NAc axons can be controlled by at least two distinct mechanisms: 1) action potentials propagating from DA cell bodies in the ventral tegmental area (VTA), and 2) activation of {beta}2* nicotinic receptors by local cholinergic interneurons (CINs). How CIN activity contributes to NAc DA dynamics in behaving animals remains unclear. We monitored DA release in the NAc Core of awake, unrestrained rats while simultaneously monitoring or manipulating CIN activity at the same location. CIN stimulation rapidly evoked DA release, and in contrast to slice preparations, this DA release showed no indication of short-term depression or receptor desensitization. The sound of food delivery evoked a brief joint increase in CIN population activity and DA release, with a second joint increase as rats approached the food. In an operant task, we observed fast ramps in CIN activity during approach behaviors, either to start the trial or to collect rewards. These CIN ramps co-occurred with DA release ramps, without corresponding changes in the firing of VTA DA neurons. Finally, we examined the effects of blocking CIN influence over DA release through local NAc infusion of DH{beta}E, a selective antagonist of [Formula] nicotinic receptors. DH{beta}E dose-dependently interfered with motivated approach decisions, mimicking the effects of a DA antagonist. Our results support a key influence of CINs over motivated behavior via the local regulation of DA release.
]]></description>
<dc:creator>Mohebi, A.</dc:creator>
<dc:creator>Collins, V. L.</dc:creator>
<dc:creator>Berke, J. D.</dc:creator>
<dc:date>2022-11-06</dc:date>
<dc:identifier>doi:10.1101/2022.11.06.515335</dc:identifier>
<dc:title><![CDATA[Cholinergic Interneurons Drive Motivation by Promoting Dopamine Release in the Nucleus Accumbens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.07.515446v1?rss=1">
<title>
<![CDATA[
Cell-subtype specific effects of genetic variation in the aging and Alzheimer cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.07.515446v1?rss=1</link>
<description><![CDATA[
The relationship between genetic variation and gene expression in individual brain cell types and subtypes has remained elusive. Here, we generated single-nucleus RNA sequencing data from the dorsolateral prefrontal cortex of 424 individuals of advanced age; analyzing 1.5 million nuclear transcriptomes, we assessed the effect of genetic variants on RNA expression in cis (cis-eQTL) for 7 cell types and 81 cell subtypes. This effort identified 10,004 eGenes at the cell type level and 8,138 eGenes at the cell subtype level. Many eGenes are only detected within cell subtypes. A new variant influences APOE expression only in microglia and is associated with greater cerebral amyloid angiopathy but not Alzheimer pathology, accounting for the effect of APOE{varepsilon}4, providing mechanistic insights into both pathologies. While eQTLs are readily detected, only a TMEM106B variant robustly affects the proportion of cell subtypes. Integration of these results with GWAS highlighted the targeted cell type and likely causal gene within susceptibility loci for Alzheimers, Parkinsons, schizophrenia, and educational attainment.
]]></description>
<dc:creator>Fujita, M.</dc:creator>
<dc:creator>Gao, Z.</dc:creator>
<dc:creator>Zeng, L.</dc:creator>
<dc:creator>McCabe, C.</dc:creator>
<dc:creator>White, C. C.</dc:creator>
<dc:creator>Ng, B.</dc:creator>
<dc:creator>Green, G. S.</dc:creator>
<dc:creator>Rozenblatt-Rosen, O.</dc:creator>
<dc:creator>Phillips, D.</dc:creator>
<dc:creator>Amir-Zilberstein, L.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Pearse, R. V.</dc:creator>
<dc:creator>Khan, A.</dc:creator>
<dc:creator>Vardarajan, B. N.</dc:creator>
<dc:creator>Kiryluk, K.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Klein, H.-U.</dc:creator>
<dc:creator>Wang, G.</dc:creator>
<dc:creator>Regev, A.</dc:creator>
<dc:creator>Habib, N.</dc:creator>
<dc:creator>Schneider, J. A.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Young-Pearse, T.</dc:creator>
<dc:creator>Mostafavi, S.</dc:creator>
<dc:creator>Bennett, D. A.</dc:creator>
<dc:creator>Menon, V.</dc:creator>
<dc:creator>De Jager, P. L.</dc:creator>
<dc:date>2022-11-08</dc:date>
<dc:identifier>doi:10.1101/2022.11.07.515446</dc:identifier>
<dc:title><![CDATA[Cell-subtype specific effects of genetic variation in the aging and Alzheimer cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.08.513062v1?rss=1">
<title>
<![CDATA[
Dynamic CD8+ T cell responses to cancer immunotherapy in human regional lymph nodes are disrupted by metastasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.08.513062v1?rss=1</link>
<description><![CDATA[
CD8+ T cell responses are critical for anti-tumor immunity. While extensively profiled in the tumor microenvironment (TME), recent studies in mice identified responses in lymph nodes (LN) as essential; however, the role of LN in human cancer patients remains unknown. We examined CD8+ T cells in human head and neck squamous cell carcinomas, regional LN, and blood using mass cytometry, single-cell genomics, and multiplexed ion beam imaging. We identified progenitor exhausted CD8+ T cells (Tpex) that were abundant in uninvolved LN and clonally related to terminally exhausted cells in the TME. After anti-PD-L1 immunotherapy, Tpex in uninvolved LN reduced in frequency but localized near dendritic cells and proliferating intermediate-exhausted CD8+ T cells (Tex-int), consistent with activation and differentiation. LN responses coincided with increased circulating Tex-int. In metastatic LN, these response hallmarks were impaired by immunosuppressive cellular niches. Our results identify important roles for LN in anti-tumor immune responses in humans.
]]></description>
<dc:creator>Rahim, M. K.</dc:creator>
<dc:creator>Okholm, T. L. H.</dc:creator>
<dc:creator>Jones, K. B.</dc:creator>
<dc:creator>McCarthy, E. E.</dc:creator>
<dc:creator>Liu, C. C.</dc:creator>
<dc:creator>Yee, J. L.</dc:creator>
<dc:creator>Tamaki, S. J.</dc:creator>
<dc:creator>Marquez, D. M.</dc:creator>
<dc:creator>Tenvooren, I.</dc:creator>
<dc:creator>Wai, K.</dc:creator>
<dc:creator>Cheung, A.</dc:creator>
<dc:creator>Davidson, B. R.</dc:creator>
<dc:creator>Johri, V.</dc:creator>
<dc:creator>Samad, B.</dc:creator>
<dc:creator>O'Gorman, W. E.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Angelo, M.</dc:creator>
<dc:creator>Fong, L.</dc:creator>
<dc:creator>Algazi, A. P.</dc:creator>
<dc:creator>Ha, P.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:date>2022-11-08</dc:date>
<dc:identifier>doi:10.1101/2022.11.08.513062</dc:identifier>
<dc:title><![CDATA[Dynamic CD8+ T cell responses to cancer immunotherapy in human regional lymph nodes are disrupted by metastasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.09.515836v1?rss=1">
<title>
<![CDATA[
memento: Generalized differential expression analysis of single-cell RNA-seq with method of moments estimation and efficient resampling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.09.515836v1?rss=1</link>
<description><![CDATA[
Differential expression analysis of scRNA-seq data is central for characterizing how experimental factors affect the distribution of gene expression. However, it remains challenging to distinguish biological and technical sources of cell-cell variability and to assess the statistical significance of quantitative comparisons between groups of cells. We introduce memento to address these limitations and enable accurate and efficient differential expression analysis of the mean, variability, and gene correlation from scRNA-seq. We used memento to analyze 70,000 tracheal epithelial cells to identify interferon response genes with distinct variability and correlation patterns, 160,000 T cells perturbed with CRISPR-Cas9 to reconstruct gene-regulatory networks that control T cell activation, and 1.2 million PMBCs to map cell-type-specific cis expression quantitative trait loci (eQTLs). In all cases, memento identified more significant and reproducible differences in mean expression but also identified differences in variability and gene correlation that suggest distinct modes of transcriptional regulation imparted by cytokines, genetic perturbations, and natural genetic variation. These results demonstrate memento as a first-in-class method for the quantitative comparisons of scRNA-seq data scalable to millions of cells and thousands of samples.
]]></description>
<dc:creator>Kim, M. C.</dc:creator>
<dc:creator>Gate, R. E.</dc:creator>
<dc:creator>Lee, D. S.</dc:creator>
<dc:creator>Chun, A. L.</dc:creator>
<dc:creator>Gordon, E.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Ntranos, V.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:date>2022-11-10</dc:date>
<dc:identifier>doi:10.1101/2022.11.09.515836</dc:identifier>
<dc:title><![CDATA[memento: Generalized differential expression analysis of single-cell RNA-seq with method of moments estimation and efficient resampling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.11.516204v1?rss=1">
<title>
<![CDATA[
Core Defense Hotspots within Pseudomonas aeruginosa are a consistent and rich source of anti-phage defense systems 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.11.516204v1?rss=1</link>
<description><![CDATA[
Bacteria use a diverse arsenal of anti-phage immune systems, including CRISPR-Cas and restriction enzymes. Identifying the full defense repertoire of a given species is still challenging, however. Here, we developed a computational tool to broadly identify anti-phage systems, which was applied to >180,000 genomes available on NCBI, revealing Pseudomonas aeruginosa to possess the most diverse anti-phage arsenal of any species with >200 sequenced genomes. Using network analysis to identify the common neighbors of anti-phage systems, we surprisingly identified two highly conserved core defense hotspot loci (cDHS1 and cDHS2). Across more than 1,000 P. aeruginosa strains, cDHS1 is up to 224 kb (mean: 34 kb) with varied arrangements of at least 31 immune systems while cDHS2 has 24 distinct systems (mean: 15.4 kb). cDHS1/2 are present in most P. aeruginosa isolates, in contrast to highly variable mobile DHSs. Most cDHS genes are of unknown function potentially representing new anti-phage systems, which we validated by identifying a novel anti-phage system (Shango) commonly encoded in cDHS1. Identification of core gene markers that flank immune islands could be a simple approach for immune system discovery and may represent popular landing spots for diverse MGEs carrying anti-phage systems.
]]></description>
<dc:creator>Johnson, M. C.</dc:creator>
<dc:creator>Laderman, E.</dc:creator>
<dc:creator>Huiting, E. C.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Davidson, A. R.</dc:creator>
<dc:creator>Bondy-Denomy, J. R.</dc:creator>
<dc:date>2022-11-11</dc:date>
<dc:identifier>doi:10.1101/2022.11.11.516204</dc:identifier>
<dc:title><![CDATA[Core Defense Hotspots within Pseudomonas aeruginosa are a consistent and rich source of anti-phage defense systems]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.11.516218v1?rss=1">
<title>
<![CDATA[
Circuit dissection and functional validation of a cross-species emotional biomarker 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.11.516218v1?rss=1</link>
<description><![CDATA[
Emotional responses arise from limbic circuits including the hippocampus and amygdala. In the human brain, beta-frequency communication between these structures correlates with self-reported mood and anxiety. However, both the mechanism and significance of this biomarker as a readout vs. driver of emotional state remain unknown. Here we show that beta-frequency communication between the ventral hippocampus and basolateral amygdala also predicts anxiety-related behavior in mice on both long timescales ([~]30 min) and immediately preceding behavioral choices. Genetically encoded voltage indicators reveal that this biomarker reflects synchronization between somatostatin interneurons across both structures. Indeed, synchrony between these neurons dynamically predicts approach vs. avoidance, and optogenetically shifting this synchronization by just 25 msec is sufficient to bidirectionally modulate anxiety-related behaviors. Thus, back-translation establishes a human biomarker as a causal determinant (not just predictor) of emotional state, revealing a novel mechanism whereby interregional synchronization that is frequency-, phase- and cell type-specific controls anxiety processing.
]]></description>
<dc:creator>Jackson, A.</dc:creator>
<dc:creator>Cohen, J.</dc:creator>
<dc:creator>Phensy, A.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Dawes, H.</dc:creator>
<dc:creator>Sohal, V.</dc:creator>
<dc:date>2022-11-11</dc:date>
<dc:identifier>doi:10.1101/2022.11.11.516218</dc:identifier>
<dc:title><![CDATA[Circuit dissection and functional validation of a cross-species emotional biomarker]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.11.515474v1?rss=1">
<title>
<![CDATA[
An RFX transcription factor regulated ciliogenesis in the progenitors of choanoflagellates and animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.11.515474v1?rss=1</link>
<description><![CDATA[
Little is known about the origins of the transcriptional modules that coordinate cell-type specific functions in animals. The controlled expression of one cellular feature - the cilium - was likely critical during early animal evolution. Two key transcription factors, RFX and FoxJ1, coordinate ciliogenesis in animals but are absent from the genomes of most other ciliated eukaryotes, raising the question of how the transcriptional regulation of ciliogenesis has evolved. To reconstruct the evolution of the RFX/FoxJ1 transcriptional module and its role in the regulation of ciliogenesis, we investigated RFX and FoxJ1 function in one of the closest living relatives of animals, the choanoflagellate Salpingoeca rosetta. Targeted disruption of the S. rosetta RFX homolog cRFXa resulted in delayed cell proliferation and aberrant ciliogenesis, marked by the collapse and resorption of nascent cilia. Ciliogenesis genes and foxJ1 were significantly down-regulated in cRFXa mutants, consistent with a pre-animal ancestry for this transcriptional module. We also found that cRFXa protein preferentially binds to a sequence motif that is enriched in the promoters of S. rosetta ciliary genes and matches the sequence motif bound by animal RFX proteins. These findings suggest that RFX coordinated ciliogenesis before the divergence of animals and choanoflagellates, and that the deployment of this module may have provided a mechanism to differentiate ciliated and non-ciliated cell types in early animal evolution.
]]></description>
<dc:creator>Coyle, M.</dc:creator>
<dc:creator>Tajima, A.</dc:creator>
<dc:creator>Leon, F.</dc:creator>
<dc:creator>Choksi, S. P.</dc:creator>
<dc:creator>Yang, A.</dc:creator>
<dc:creator>Espinoza, S.</dc:creator>
<dc:creator>Hughes, T. R.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:creator>Booth, D. S.</dc:creator>
<dc:creator>King, N.</dc:creator>
<dc:date>2022-11-14</dc:date>
<dc:identifier>doi:10.1101/2022.11.11.515474</dc:identifier>
<dc:title><![CDATA[An RFX transcription factor regulated ciliogenesis in the progenitors of choanoflagellates and animals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.15.516588v1?rss=1">
<title>
<![CDATA[
Pseudomonas aeruginosa mechanosensing controls cell polarity during twitching by activating two antagonistic response regulators 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.15.516588v1?rss=1</link>
<description><![CDATA[
The opportunistic pathogen Pseudomonas aeruginosa adapts to solid surfaces to enhance virulence and infect its host. Type IV pili (T4P), long and thin filaments that power surface-specific twitching motility, allow single cells to mechanosense surfaces. For example, cells sense T4P attachment to control the direction of twitching motility. In this process, they establish a local positive feedback that polarizes T4P distribution to the sensing pole. A complex chemotaxis-like system called Chp mediates this response. The signalling mechanism allowing for transduction of this spatially-resolved signal is however unresolved. Here we demonstrate that the two Chp response regulators PilG and PilH enable dynamic cell polarization by coupling their antagonistic functions on T4P extension. By precisely quantifying the localization of fluorescent protein fusions, we show that PilG polarizes in response to mechanosensing through phosphorylation by the histidine kinase ChpA. We find that PilH is not inherently required for reversals. However, PilH activation is necessary to break the local positive feedback established by PilG so that forward-twitching cells can reverse. To spatially resolve mechanical signals, Chp thus locally transduces signals with a main output response regulator, PilG. To respond to signal changes, Chp uses its second regulator PilH to break the local feedback. By identifying the molecular functions of two response regulators that dynamically control cell polarization, our work provides a rationale for the diversity of architectures often found in non-canonical chemotaxis systems.
]]></description>
<dc:creator>Kuhn, M. J.</dc:creator>
<dc:creator>Macmillan, H.</dc:creator>
<dc:creator>Tala, L.</dc:creator>
<dc:creator>Inclan, Y.</dc:creator>
<dc:creator>Patino, R.</dc:creator>
<dc:creator>Pierrat, X.</dc:creator>
<dc:creator>Al-Mayyah, Z.</dc:creator>
<dc:creator>Engel, J. N.</dc:creator>
<dc:creator>Persat, A.</dc:creator>
<dc:date>2022-11-15</dc:date>
<dc:identifier>doi:10.1101/2022.11.15.516588</dc:identifier>
<dc:title><![CDATA[Pseudomonas aeruginosa mechanosensing controls cell polarity during twitching by activating two antagonistic response regulators]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.13.516350v1?rss=1">
<title>
<![CDATA[
Targeted high throughput mutagenesis of the human spliceosome reveals its in vivo operating principles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.13.516350v1?rss=1</link>
<description><![CDATA[
The spliceosome is a staggeringly complex machine comprising, in humans, 5 snRNAs and >150 proteins. We scaled haploid CRISPR-Cas9 base editing to target the entire human spliceosome and interrogated the mutants using the U2 snRNP/SF3b inhibitor, pladienolide B. Hypersensitive substitutions define functional sites in the U1/U2-containing A-complex but also in components that act as late as the second chemical step after SF3b is dissociated. Viable resistance substitutions map not only to the pladienolide B binding site but also to the G-patch (ATPase activator) domain of SUGP1, which lacks orthologs in yeast. We used these mutants and biochemical approaches to identify the spliceosomal disassemblase DHX15/hPrp43 as the ATPase ligand for SUGP1. These and other data support a model in which SUGP1 promotes splicing fidelity by triggering early spliceosome disassembly in response to kinetic blocks. Our approach provides a template for the analysis of essential cellular machines in humans.
]]></description>
<dc:creator>Beusch, I.</dc:creator>
<dc:creator>Rao, B.</dc:creator>
<dc:creator>Studer, M.</dc:creator>
<dc:creator>Luhovska, T.</dc:creator>
<dc:creator>Sukyte, V.</dc:creator>
<dc:creator>Lei, S.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>SeGraves, E.</dc:creator>
<dc:creator>Burlingame, A.</dc:creator>
<dc:creator>Jonas, S.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:date>2022-11-16</dc:date>
<dc:identifier>doi:10.1101/2022.11.13.516350</dc:identifier>
<dc:title><![CDATA[Targeted high throughput mutagenesis of the human spliceosome reveals its in vivo operating principles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.15.516217v1?rss=1">
<title>
<![CDATA[
Uptake of tumor-derived microparticles induces metabolic reprogramming of macrophages in the early metastatic lung. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.15.516217v1?rss=1</link>
<description><![CDATA[
The formation of a pre-metastatic niche is a critical step during the metastatic spread of cancer. One way by which primary tumors prime host cells at future metastatic sites is through local shedding of tumor-derived microparticles as a consequence of vascular sheer flow. However, it remains unclear how the uptake of such particles by resident immune cells affects their phenotype and function. Here we show that ingestion of tumor-derived microparticles by macrophages induces a rapid metabolic and phenotypic switch that is characterized by enhanced mitochondrial mass and function, increased oxidative phosphorylation and upregulation of cellular adhesion molecules resulting in reduced motility in the early metastatic lung. We show that this reprogramming event is dependent on signaling through the mTORC1, but not mTORC2 pathway, and is unique to uptake of tumor-derived microparticles. Together, these data support a mechanism by which uptake of tumor-derived microparticles induces reprogramming of macrophages to shape their fate and function in the early metastatic lung.
]]></description>
<dc:creator>Kersten, K.</dc:creator>
<dc:creator>You, R.</dc:creator>
<dc:creator>Liang, S.</dc:creator>
<dc:creator>Tharp, K.</dc:creator>
<dc:creator>Pollack, J.</dc:creator>
<dc:creator>Weaver, V. M.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Headley, M. B.</dc:creator>
<dc:date>2022-11-17</dc:date>
<dc:identifier>doi:10.1101/2022.11.15.516217</dc:identifier>
<dc:title><![CDATA[Uptake of tumor-derived microparticles induces metabolic reprogramming of macrophages in the early metastatic lung.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.18.517047v1?rss=1">
<title>
<![CDATA[
Innate immune response to SARS-1 CoV-2 infection contributes to neuronal damage in human iPSC-derived peripheral neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.18.517047v1?rss=1</link>
<description><![CDATA[
Severe acute respiratory coronavirus 2 (SARS-CoV-2) infection causes neurological disease in some patients suggesting that infection can affect both the peripheral and central nervous system (PNS and CNS, respectively). It is not clear whether the outcome of SARS-CoV-2 infection of PNS and CNS neurons is similar, and which are the key factors that cause neurological disease: SARS-CoV-2 infection or the subsequent immune response. Here, we addressed these questions by infecting human induced-pluripotent stem cell-derived CNS and PNS neurons with the {beta} strain of SARS-CoV-2. Our results show that SARS-CoV-2 infects PNS neurons more efficiently than CNS neurons, despite lower expression levels of angiotensin converting enzyme 2. Infected PNS neurons produced interferon {lambda}1, several interferon stimulated genes and proinflammatory cytokines. They also displayed neurodegenerative-like alterations, as indicated by increased levels of sterile alpha and Toll/interleukin receptor motif-containing protein 1, amyloid precursor protein and -synuclein and lower levels of nicotinamide mononucleotide adenylyltransferase 2 and {beta}-III-tubulin. Interestingly, blockade of the Janus kinase and signal transducer and activator of transcription pathway by Ruxolitinib did not increase SARS-CoV-2 infection, but reduced neurodegeneration, suggesting that an exacerbated neuronal innate immune response contributes to pathogenesis in the PNS.
]]></description>
<dc:creator>Passos, V.</dc:creator>
<dc:creator>Henkel, L. M.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Zapatero-Belinchon, F. J.</dc:creator>
<dc:creator>Moeller, R.</dc:creator>
<dc:creator>Sun, G.</dc:creator>
<dc:creator>Waltl, I.</dc:creator>
<dc:creator>Ritter, B.</dc:creator>
<dc:creator>Kropp, K. A.</dc:creator>
<dc:creator>Zhu, S.</dc:creator>
<dc:creator>Deleidi, M.</dc:creator>
<dc:creator>Kalinke, U.</dc:creator>
<dc:creator>Hoeglinger, G.</dc:creator>
<dc:creator>Gerold, G.</dc:creator>
<dc:creator>Wegner, F.</dc:creator>
<dc:creator>Viejo-Borbolla, A.</dc:creator>
<dc:date>2022-11-18</dc:date>
<dc:identifier>doi:10.1101/2022.11.18.517047</dc:identifier>
<dc:title><![CDATA[Innate immune response to SARS-1 CoV-2 infection contributes to neuronal damage in human iPSC-derived peripheral neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.18.517147v1?rss=1">
<title>
<![CDATA[
Tracing the origin of pathologic pulmonary fibroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.18.517147v1?rss=1</link>
<description><![CDATA[
Fibroblasts substantially remodel extracellular matrix (ECM) in response to tissue injury and generate fibrotic scars in chronic diseases. Recent studies have identified diverse fibroblast subsets in healthy and injured tissues. However, the origin(s) and functional importance of injury-induced fibroblast lineages remain unclear. Here we show that alveolar fibroblasts, which provide a niche for maintaining alveolar type 2 cells in uninjured lungs, are the dominant source of multiple emergent fibroblast subsets that sequentially arise to facilitate fibrosis after lung injury. We demonstrate that Cthrc1+ fibroblasts, which express the highest levels of ECM proteins at injured sites, are effector cells for fibrogenesis using a novel mouse tool, Cthrc1-CreER. We use another novel mouse tool, Scube2-CreER, that uniquely targets alveolar fibroblasts, to reveal that alveolar fibroblasts are the dominant origin for multiple fibroblast subsets that emerge after lung injury. Pseudotime and in vitro analysis suggest that inflammatory cytokines initially induce chemokine-producing inflammatory fibroblasts from alveolar fibroblasts, which can differentiate into Cthrc1+ fibrotic fibroblasts in response to TGF-{beta}. We identify similar fibroblast lineages from scRNA-seq in human pulmonary fibrosis. These results elucidate the pathologic fibroblast lineage development in response to lung injury and suggest that targeting key steps in transitions among these subsets could provide novel strategies for the treatment of fibrotic diseases.
]]></description>
<dc:creator>Tsukui, T.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:date>2022-11-18</dc:date>
<dc:identifier>doi:10.1101/2022.11.18.517147</dc:identifier>
<dc:title><![CDATA[Tracing the origin of pathologic pulmonary fibroblasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.20.517219v1?rss=1">
<title>
<![CDATA[
Human endometrial KISS1R inhibits stromal cell decidualization in a manner associated with a reduction in ESR1 levels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.20.517219v1?rss=1</link>
<description><![CDATA[
Defective endometrial stromal cell decidualization is a major cause of recurrent implantation failure (RIF), a condition with a prevalence of [~]15%. To treat RIF, a stronger understanding of the endometrial factors that regulate decidualization is required. Here we studied the role of the kisspeptin receptor (KISS1R) in regulating human endometrial stromal cell (HESC) decidualization. Our data revealed KISS1R inhibits HESC decidualization in vitro in a manner associated with a striking reduction in ESR1 protein levels. To determine whether KISSR inhibition of decidualization results from reduced ESR1 levels we expressed the dominant negative ESR1-46 isoform in decidualizing HESCs. We found that expression of ESR1-46 in decidualizing HESCs ablated the expression of ESR1-66 and ESR1-54 isomers, and blocked decidualization. Interestingly, when ESR1-64 was co-expressed with ESR1-46, ESR1-66 and ESR1-54 expression was restored and decidualization was rescued. Taken together, these results suggest that KISS1R inhibits HESC decidualization by downregulating ESR1 levels. Based on our findings, we suggest that by inhibiting HESC decidualization, KISS1R regulates the depth of embryo invasion of the stroma, a requirement for a successful pregnancy.
]]></description>
<dc:creator>Schaefer, J.</dc:creator>
<dc:creator>Chadchan, S.</dc:creator>
<dc:creator>George, A.</dc:creator>
<dc:creator>Roan, N.</dc:creator>
<dc:creator>Bhattacharya, M.</dc:creator>
<dc:creator>Kommagaini, R.</dc:creator>
<dc:creator>Babwah, A.</dc:creator>
<dc:date>2022-11-20</dc:date>
<dc:identifier>doi:10.1101/2022.11.20.517219</dc:identifier>
<dc:title><![CDATA[Human endometrial KISS1R inhibits stromal cell decidualization in a manner associated with a reduction in ESR1 levels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.20.517228v1?rss=1">
<title>
<![CDATA[
Robust and Highly Efficient Extractions of Proteins from Bones enable Deep, High-Throughput Proteomic Quantification to Gain Insights into Bone Biology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.20.517228v1?rss=1</link>
<description><![CDATA[
Dysregulation of cell signaling in chondrocytes and in bone cells, such as osteocytes, osteoblasts, osteoclasts, and an elevated burden of senescent cells in cartilage and bone, are implicated in osteoarthritis (OA). Mass spectrometric analyses provides a crucial molecular tool-kit to understand complex signaling relationships in age-related diseases, such as OA. Here we introduce a novel mass spectrometric workflow to promote proteomic studies of bone and cartilage. This workflow uses highly specialized steps, including extensive overnight demineralization, pulverization, and incubation for 72 h in 6 M guanidine hydrochloride and EDTA, followed by proteolytic digestion. Analysis on a high-resolution Orbitrap Eclipse and Orbitrap Exploris 480 mass spectrometer using Data-Independent Acquisition (DIA) provides deep coverage of the bone proteome, and preserves post-translational modifications, such as hydroxyproline. A spectral library-free quantification strategy, directDIA, identified and quantified over 2,000 protein groups (with [&ge;] 2 unique peptides) from calcium-rich bone matrices. Key components identified were proteins of the extracellular matrix (ECM), bone-specific proteins (e.g., secreted protein acidic and cysteine rich, SPARC, and bone sialoprotein 2, IBSP), and signaling proteins (e.g., transforming growth factor beta-2, TGFB2), and lysyl oxidase homolog 2 (LOXL2), an important protein in collagen crosslinking. Post-translational modifications (PTMs) were identified without the need for specific enrichment. This includes collagen hydroxyproline modifications, chemical modifications for collagen self-assembly and network formation. Multiple senescence factors were identified, such as complement component 3 (C3) protein of the complement system and many matrix metalloproteinases, that might be monitored during age-related bone disease progression. Our innovative workflow yields in-depth protein coverage and quantification strategies to discover underlying biological mechanisms of bone aging and to provide tools to monitor therapeutic interventions. These novel tools to monitor the bone proteome open novel horizons to investigate bone-specific diseases, many of which are age-related.
]]></description>
<dc:creator>Rose, J. P.</dc:creator>
<dc:creator>Schurman, C. A.</dc:creator>
<dc:creator>King, C. D.</dc:creator>
<dc:creator>Bons, J.</dc:creator>
<dc:creator>Burton, J. B.</dc:creator>
<dc:creator>Patel, S. K.</dc:creator>
<dc:creator>O'Broin, A.</dc:creator>
<dc:creator>Alliston, T.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:date>2022-11-20</dc:date>
<dc:identifier>doi:10.1101/2022.11.20.517228</dc:identifier>
<dc:title><![CDATA[Robust and Highly Efficient Extractions of Proteins from Bones enable Deep, High-Throughput Proteomic Quantification to Gain Insights into Bone Biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.20.517270v1?rss=1">
<title>
<![CDATA[
Systematic Approach Identifies Multiple Transcription Factor Perturbations That Rejuvenate Replicatively Aged Human Skin Fibroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.20.517270v1?rss=1</link>
<description><![CDATA[
Rejuvenation, long a quixotic dream, recently became a possibility through exciting new approaches to counteract aging. For example, parabiosis and partial reprogramming through overexpressing four stem cell transcription factors (Yamanaka factors) both rejuvenate organisms and cells1-5. We hypothesize there are many other genetic solutions to human cell rejuvenation, and some solutions may be safer and more potent than current gene targets. We set out to develop a systematic approach to identify novel genes that, when overexpressed or repressed, reprogram the global gene expression of a cell back to a younger state. Using the Hayflick model of human cell replicative aging, we performed a Perturb-seq screen of 200 transcription factors (TFs) selected through a combination of bioinformatic analysis and literature search. We identified dozens of potentially rejuvenating TFs--those that when overexpressed or repressed in late passage cells reprogrammed global gene expression patterns back to an earlier passage state. We further validated four top TF perturbations through molecular phenotyping of various aging hallmarks. Late passage cells either overexpressing EZH2 or E2F3 or repressing STAT3 or ZFX had more cell division, less senescence, improved proteostasis, and enhanced mitochondrial function. These TF perturbations led to similar downstream gene expression programs. In addition, the rejuvenating effects of these TFs were independent of telomeres. We believe our general approach for identifying rejuvenating factors can be applied to other model systems, and some of the top TF perturbations we discovered will lead to future research in novel, safer rejuvenation therapies.
]]></description>
<dc:creator>Sengstack, J.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Mobaraki, M.</dc:creator>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Deng, C.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2022-11-20</dc:date>
<dc:identifier>doi:10.1101/2022.11.20.517270</dc:identifier>
<dc:title><![CDATA[Systematic Approach Identifies Multiple Transcription Factor Perturbations That Rejuvenate Replicatively Aged Human Skin Fibroblasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.21.517375v1?rss=1">
<title>
<![CDATA[
Fragment-Based Hit Discovery via Unsupervised Learning of Fragment-Protein Complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.21.517375v1?rss=1</link>
<description><![CDATA[
The process of finding molecules that bind to a target protein is a challenging first step in drug discovery. Crystallographic fragment screening is a strategy based on elucidating binding modes of small polar compounds and then building potency by expanding or merging them. Recent advances in high-throughput crystallography enable screening of large fragment libraries, reading out dense ensembles of fragments spanning the binding site. However, fragments typically have low affinity thus the road to potency is often long and fraught with false starts. Here, we take advantage of high-throughput crystallography to reframe fragment-based hit discovery as a denoising problem - identifying significant pharmacophore distributions from a fragment ensemble amid noise due to weak binders - and employ an unsupervised machine learning method to tackle this problem. Our method screens potential molecules by evaluating whether they recapitulate those fragment-derived pharmacophore distributions. We retrospectively validated our approach on an open science campaign against SARS-CoV-2 main protease (Mpro), showing that our method can distinguish active compounds from inactive ones using only structural data of fragment-protein complexes, without any activity data. Further, we prospectively found novel hits for Mpro and the Mac1 domain of SARS-CoV-2 non-structural protein 3. More broadly, our results demonstrate how unsupervised machine learning helps interpret high throughput crystallography data to rapidly discover of potent chemical modulators of protein function.
]]></description>
<dc:creator>McCorkindale, W. J.</dc:creator>
<dc:creator>Ahel, I.</dc:creator>
<dc:creator>Barr, H.</dc:creator>
<dc:creator>Correy, G. J.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>London, N.</dc:creator>
<dc:creator>Schuller, M.</dc:creator>
<dc:creator>Shurrush, K.</dc:creator>
<dc:creator>Lee, A. A.</dc:creator>
<dc:date>2022-11-24</dc:date>
<dc:identifier>doi:10.1101/2022.11.21.517375</dc:identifier>
<dc:title><![CDATA[Fragment-Based Hit Discovery via Unsupervised Learning of Fragment-Protein Complexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.21.517399v1?rss=1">
<title>
<![CDATA[
Anti-integrin αvβ6 autoantibodies are a novel predictive biomarker in ulcerative colitis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.21.517399v1?rss=1</link>
<description><![CDATA[
Background and AimsBetter biomarkers for prediction of ulcerative colitis (UC) development and prognostication are needed. Anti-integrin v{beta}6 autoantibodies (anti-v{beta}6) have been described in UC patients. Here, we tested for the presence of anti-v{beta}6 antibodies in the pre-clinical phase of UC and studied their association with disease-related outcomes after diagnosis.

MethodsAnti-v{beta}6 were measured in 4 longitudinal serum samples collected from 82 subjects who later developed UC and 82 matched controls from a Department of Defense pre-clinical cohort (PREDICTS). In a distinct, external validation cohort (GEM), we tested 12 pre-UC subjects and 49 matched controls. Further, anti-v{beta}6 were measured in 2 incident UC cohorts (COMPASS n=55 and OSCCAR n=104) and associations between anti-v{beta}6 and UC-related outcomes were defined using Cox proportional-hazards model.

ResultsAnti-v{beta}6 were significantly higher among individuals who developed UC compared to controls up to 10 years before diagnosis in PREDICTS. The anti-v{beta}6 seropositivity was 12.2% 10 years before diagnosis and increased to 52.4% at the time of diagnosis in subjects who developed UC compared with 2.7% in controls across the 4 timepoints. Anti-v{beta}6 predicted UC development with an AUC of at least 0.8 up to 10 years before diagnosis. The presence of anti-v{beta}6 in pre-clinical UC samples was validated in the GEM cohort. Finally, high anti-v{beta}6 was associated with a composite of adverse UC-outcomes including hospitalization, disease extension, colectomy, systemic steroid use and/or escalation to biologic therapy in recently diagnosed UC.

ConclusionAnti-integrin v{beta}6 auto-antibodies precede the clinical diagnosis of UC by up to 10 years and are associated with adverse UC-related outcomes.
]]></description>
<dc:creator>Livanos, A. E.</dc:creator>
<dc:creator>Dunn, A.</dc:creator>
<dc:creator>Fischer, J.</dc:creator>
<dc:creator>Ungaro, R. C.</dc:creator>
<dc:creator>Turpin, W.</dc:creator>
<dc:creator>Lee, S.-H.</dc:creator>
<dc:creator>Rui, S.</dc:creator>
<dc:creator>Del Valle, D. M.</dc:creator>
<dc:creator>Julia, J. J.</dc:creator>
<dc:creator>Martinez-Delgado, G.</dc:creator>
<dc:creator>Riddle, M. S.</dc:creator>
<dc:creator>Murray, J. A.</dc:creator>
<dc:creator>Laird, R. M.</dc:creator>
<dc:creator>Torres, J.</dc:creator>
<dc:creator>Agrawal, M.</dc:creator>
<dc:creator>Magee, J. S.</dc:creator>
<dc:creator>Dervieux, T.</dc:creator>
<dc:creator>Gnjatic, S.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:creator>Sands, B. E.</dc:creator>
<dc:creator>Porter, C. K.</dc:creator>
<dc:creator>Croitoru, K.</dc:creator>
<dc:creator>Petralia, F.</dc:creator>
<dc:creator>Colombel, J.-F.</dc:creator>
<dc:creator>Mehandru, S.</dc:creator>
<dc:date>2022-11-24</dc:date>
<dc:identifier>doi:10.1101/2022.11.21.517399</dc:identifier>
<dc:title><![CDATA[Anti-integrin αvβ6 autoantibodies are a novel predictive biomarker in ulcerative colitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.26.517738v1?rss=1">
<title>
<![CDATA[
Kinetochore-fiber lengths are maintained locally but coordinated globally by poles in the mammalian spindle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.26.517738v1?rss=1</link>
<description><![CDATA[
At each cell division, nanometer-scale components self-organize to build a micron-scale spindle. In mammalian spindles, microtubule bundles called kinetochore-fibers attach to chromosomes and focus into spindle poles. Despite evidence suggesting that poles can set spindle length, their role remains poorly understood. In fact, many species do not have spindle poles. Here, we probe the poles contribution to mammalian spindle length, dynamics, and function by inhibiting dynein to generate spindles whose kinetochore-fibers do not focus into poles, yet maintain a metaphase steady-state length. We find that unfocused kinetochore-fibers have a mean length indistinguishable from control, but a broader length distribution, and reduced length coordination between sisters and neighbors. Further, we show that unfocused kinetochore-fibers, like control, can grow back to their steady-state length if acutely shortened by drug treatment or laser ablation: they recover their length by tuning their end dynamics, albeit slower due to their reduced baseline dynamics. Thus, kinetochore-fiber dynamics are regulated by their length, not just pole-focusing forces. Finally, we show that spindles with unfocused kinetochore-fibers can segregate chromosomes but fail to correctly do so. We propose that mammalian spindle length emerges locally from individual k-fibers while spindle poles globally coordinate k-fibers across space and time.
]]></description>
<dc:creator>Richter, M.</dc:creator>
<dc:creator>Neahring, L.</dc:creator>
<dc:creator>Tao, J.</dc:creator>
<dc:creator>Sutanto, R.</dc:creator>
<dc:creator>Cho, N. H.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2022-11-26</dc:date>
<dc:identifier>doi:10.1101/2022.11.26.517738</dc:identifier>
<dc:title><![CDATA[Kinetochore-fiber lengths are maintained locally but coordinated globally by poles in the mammalian spindle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.27.518061v1?rss=1">
<title>
<![CDATA[
Evidence that SPIO Chain Formation is Essential for High-Resolution MPI 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.27.518061v1?rss=1</link>
<description><![CDATA[
Magnetic Particle Imaging (MPI) is a noninvasive imaging modality that exploits the saturation properties of superparamagnetic iron oxide particles (SPIOs). A major thrust of MPI research aims to sharpen the magnetic resolution of biocompatible SPIOs, which will be crucial for affordable and safe clinical translation. We recently reported on a new class of MPI tracers --called superferromagnetic iron oxide nanoparticles (SFMIOs) -- which offer much sharper magnetic saturation curves. SFMIOs experimentally demonstrate 10-fold improvement in both resolution and sensitivity. However, superferromagnetism is a relatively unexplored branch of physics and the nanoscale physics and dynamics of SFMIOs remain a mystery. Here we show experimentally that chaining of SPIOs can explain SFMIOs boost in SNR and resolution. We show how concentration, viscosity, transmit amplitude, and pre-polarization time can all affect SPIO chain formation and SFMIO behavior. These experiments will inform strategies on SFMIO chemical synthesis as well as SFMIO data acquisition pulse sequences.
]]></description>
<dc:creator>Colson, C.</dc:creator>
<dc:creator>Fung, K. L. B.</dc:creator>
<dc:creator>Bryan, J.</dc:creator>
<dc:creator>Tay, Z. W.</dc:creator>
<dc:creator>Fellows, B. D.</dc:creator>
<dc:creator>Sayuujya, C.</dc:creator>
<dc:creator>Kuo, R.</dc:creator>
<dc:creator>Chandrasekharan, P.</dc:creator>
<dc:creator>Conolly, S. M.</dc:creator>
<dc:date>2022-11-28</dc:date>
<dc:identifier>doi:10.1101/2022.11.27.518061</dc:identifier>
<dc:title><![CDATA[Evidence that SPIO Chain Formation is Essential for High-Resolution MPI]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.27.518106v1?rss=1">
<title>
<![CDATA[
Global Endometrial DNA Multi-omics Analysis Reveals Insights into mQTL Regulation and Associated Endometriosis Disease Risk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.27.518106v1?rss=1</link>
<description><![CDATA[
Endometriosis is a leading cause of pain and infertility affecting millions of women globally. Identifying biologic and genetic effects on DNA methylation (DNAm) in endometrium increases understanding of mechanisms that influence gene regulation predisposing to endometriosis and offers an opportunity for novel therapeutic target discovery. Herein, we characterize variation in endometrial DNAm and its association with menstrual cycle phase, endometriosis, and genetic variants through analysis of genome-wide genotype data and methylation at 759,345 DNAm sites in endometrial samples from 984 deeply-phenotyped participants. We identify significant differences in DNAm profiles between menstrual cycle phases and at four DNAm sites between stage III/IV endometriosis and controls. We estimate that 15.4% of the variation in endometriosis is captured by DNAm, and identify DNAm networks associated with endometriosis. DNAm quantitative trait locus (mQTL) analysis identified 118,185 independent cis-mQTL including some tissue-specific effects. We find significant differences in DNAm profiles between endometriosis sub- phenotypes and a significant association between genetic regulation of methylation in endometrium and disease risk, providing functional evidence for genomic targets contributing to endometriosis risk and pathogenesis.
]]></description>
<dc:creator>Mortlock, S.</dc:creator>
<dc:creator>Houshdaran, S.</dc:creator>
<dc:creator>Kosti, I.</dc:creator>
<dc:creator>Rahmioglu, N.</dc:creator>
<dc:creator>Nezhat, C.</dc:creator>
<dc:creator>Vitonis, A. F.</dc:creator>
<dc:creator>Andrews, S. V.</dc:creator>
<dc:creator>Grosjean, P.</dc:creator>
<dc:creator>Paranjpe, M.</dc:creator>
<dc:creator>Horne, A. W.</dc:creator>
<dc:creator>Jacoby, A.</dc:creator>
<dc:creator>Lager, J.</dc:creator>
<dc:creator>Opoku-Anane, J.</dc:creator>
<dc:creator>Vo, K. C.</dc:creator>
<dc:creator>Manvelyan, E.</dc:creator>
<dc:creator>Sen, S.</dc:creator>
<dc:creator>Ghukasyan, Z.</dc:creator>
<dc:creator>Collins, F.</dc:creator>
<dc:creator>Santamaria, X.</dc:creator>
<dc:creator>Saunders, P.</dc:creator>
<dc:creator>Kober, K.</dc:creator>
<dc:creator>McRae, A. F.</dc:creator>
<dc:creator>Terry, K. L.</dc:creator>
<dc:creator>Vallve-Juanico, J.</dc:creator>
<dc:creator>Becker, C.</dc:creator>
<dc:creator>Rogers, P. A. W.</dc:creator>
<dc:creator>Irwin, J. C.</dc:creator>
<dc:creator>Zondervan, K.</dc:creator>
<dc:creator>Montgomery, G. W.</dc:creator>
<dc:creator>Missmer, S.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Giudice, L.</dc:creator>
<dc:date>2022-11-28</dc:date>
<dc:identifier>doi:10.1101/2022.11.27.518106</dc:identifier>
<dc:title><![CDATA[Global Endometrial DNA Multi-omics Analysis Reveals Insights into mQTL Regulation and Associated Endometriosis Disease Risk]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.28.516756v1?rss=1">
<title>
<![CDATA[
The connectome of an insect brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.28.516756v1?rss=1</link>
<description><![CDATA[
Brains contain networks of interconnected neurons, so knowing the network architecture is essential for understanding brain function. We therefore mapped the synaptic-resolution connectome of an insect brain (Drosophila larva) with rich behavior, including learning, value-computation, and action-selection, comprising 3,013 neurons and 544,000 synapses. We characterized neuron-types, hubs, feedforward and feedback pathways, and cross-hemisphere and brain-nerve cord interactions. We found pervasive multisensory and interhemispheric integration, highly recurrent architecture, abundant feedback from descending neurons, and multiple novel circuit motifs. The brains most recurrent circuits comprised the input and output neurons of the learning center. Some structural features, including multilayer shortcuts and nested recurrent loops, resembled powerful machine learning architectures. The identified brain architecture provides a basis for future experimental and theoretical studies of neural circuits.

One-Sentence SummaryWe generated a synaptic-resolution brain connectome and characterized its connection types, neuron types, and circuit motifs.
]]></description>
<dc:creator>Winding, M.</dc:creator>
<dc:creator>Pedigo, B. D.</dc:creator>
<dc:creator>Barnes, C. L.</dc:creator>
<dc:creator>Patsolic, H. G.</dc:creator>
<dc:creator>Park, Y.</dc:creator>
<dc:creator>Kazimiers, T.</dc:creator>
<dc:creator>Fushiki, A.</dc:creator>
<dc:creator>Andrade, I. V.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Valdes-Aleman, J.</dc:creator>
<dc:creator>Khandelwal, A.</dc:creator>
<dc:creator>Randel, N.</dc:creator>
<dc:creator>Barsotti, E.</dc:creator>
<dc:creator>Correia, A.</dc:creator>
<dc:creator>Fetter, R. D.</dc:creator>
<dc:creator>Hartenstein, V.</dc:creator>
<dc:creator>Priebe, C. E.</dc:creator>
<dc:creator>Vogelstein, J. T.</dc:creator>
<dc:creator>Cardona, A.</dc:creator>
<dc:creator>Zlatic, M.</dc:creator>
<dc:date>2022-11-28</dc:date>
<dc:identifier>doi:10.1101/2022.11.28.516756</dc:identifier>
<dc:title><![CDATA[The connectome of an insect brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.29.518452v1?rss=1">
<title>
<![CDATA[
Translation-dependent downregulation of Cas12a mRNA by an anti-CRISPR protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.29.518452v1?rss=1</link>
<description><![CDATA[
Bacteria have evolved multiple defense systems, including CRISPR-Cas, to cleave the DNA of phage and mobile genetic elements (MGE). In turn, phage have evolved anti-CRISPR (Acr) proteins that use novel and co-opted mechanisms to block DNA binding or cleavage. Here, we report that an anti-CRISPR (AcrVA2) unexpectedly inhibits Cas12a biogenesis by triggering translation-dependent destruction of its mRNA. AcrVA2 specifically clears the mRNA of Cas12a by recognizing and binding its N-terminal polypeptide. Mutating conserved N-terminal amino acids in Cas12a prevents binding and inhibition by AcrVA2 but also decreases Cas12a anti-phage activity. This mechanism therefore enables AcrVA2 to specifically inhibit divergent Cas12a orthologs while constraining its ability to escape inhibition. AcrVA2 homologs are found on diverse MGEs across numerous bacterial classes, typically in the absence of Cas12a, suggesting that this protein family may induce similar molecular outcomes against other targets. These findings reveal a new gene regulatory strategy in bacteria and create opportunities for polypeptide-specific gene regulation in prokaryotes and beyond.
]]></description>
<dc:creator>Marino, N. D.</dc:creator>
<dc:creator>Talaie, A.</dc:creator>
<dc:creator>Carion, H.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Silas, S.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2022-11-30</dc:date>
<dc:identifier>doi:10.1101/2022.11.29.518452</dc:identifier>
<dc:title><![CDATA[Translation-dependent downregulation of Cas12a mRNA by an anti-CRISPR protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.29.518421v1?rss=1">
<title>
<![CDATA[
Acute adenoviral cardiac infection elicits an arrhythmogenic substrate prior to inflammatory myocardial remodeling and myocarditis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.29.518421v1?rss=1</link>
<description><![CDATA[
BackgroundViral cardiac infection represents a significant clinical challenge encompassing several etiological agents, disease stages, complex presentation, and a resulting lack of mechanistic understanding. Myocarditis is a major cause of sudden cardiac death in young adults, where current knowledge in the field is dominated by later disease phases, and pathological immune responses. However, little is known regarding how infection can acutely induce an arrhythmogenic substrate prior to significant immune responses. Adenovirus is a leading cause of myocarditis, but due to species-specificity, models of infection are lacking and it is not understood how adenoviral infection may underlie sudden cardiac arrest. Mouse Adenovirus Type-3 (MAdV-3) was previously reported as cardiotropic, yet has not been utilized to understand mechanisms of cardiac infection and pathology.

MethodsWe have developed MAdV-3 infection as a model to investigate acute cardiac infection and molecular alterations to the infected heart prior to an appreciable immune response or gross cardiomyopathy.

ResultsOptical mapping of infected hearts exposes decreases in conduction velocity concomitant with increased Cx43Ser368 phosphorylation, a residue known to regulate gap junction function. Hearts from animals harboring a phospho-null mutation at Cx43Ser368 are protected against MAdV-3 induced conduction velocity slowing. Additional to gap junction alterations, patch clamping of MAdV-3-infected adult mouse ventricular cardiomyocytes reveals prolonged action potential duration as a result of decreased IK1 and IKs current density. Turning to human systems, we find human adenovirus type-5 (HAdV-5) increases phosphorylation of Cx43Ser368 and disrupts synchrony in human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs), indicating common mechanisms with our mouse whole heart and adult cardiomyocyte data.

ConclusionsTogether, these findings demonstrate that adenoviral infection creates an arrhythmogenic substrate through direct targeting of gap junction and ion channel function in the heart. Such alterations are known to precipitate arrhythmias and likely contribute to sudden cardiac death in acutely infected patients.
]]></description>
<dc:creator>Padget, R. L.</dc:creator>
<dc:creator>Blair, G. A.</dc:creator>
<dc:creator>North, M. D.</dc:creator>
<dc:creator>Zeitz, M. J.</dc:creator>
<dc:creator>Tanenbaum, M. T.</dc:creator>
<dc:creator>King, D. R.</dc:creator>
<dc:creator>Hoeker, G. S.</dc:creator>
<dc:creator>Swanger, S. A.</dc:creator>
<dc:creator>Poelzing, S.</dc:creator>
<dc:creator>Smyth, J. W.</dc:creator>
<dc:date>2022-12-02</dc:date>
<dc:identifier>doi:10.1101/2022.11.29.518421</dc:identifier>
<dc:title><![CDATA[Acute adenoviral cardiac infection elicits an arrhythmogenic substrate prior to inflammatory myocardial remodeling and myocarditis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.01.518779v1?rss=1">
<title>
<![CDATA[
An ESCRT-dependent pathway of Nuclear and Cytoplasmic Spatial PQC is coordinated at Nuclear Vacuolar Junctions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.01.518779v1?rss=1</link>
<description><![CDATA[
Effective Protein Quality Control (PQC), essential for cellular health, relies on spatial sequestration of misfolded proteins into defined inclusions. Here we elucidate the coordination of nuclear and cytoplasmic spatial PQC. While cytoplasmic misfolded proteins concentrate in a cytoplasmic, perinuclear Juxta Nuclear Quality control compartment (JUNQ), nuclear misfolded proteins sequester into a perinucleolar IntraNuclear Quality control compartment (INQ). Particle tracking reveals the INQ and JUNQ converge to face each other across the nuclear envelope at a site proximal to the Nuclear-Vacuolar Junction (NVJ) marked by perinuclear ESCRT-II/-III protein Chm7. Strikingly, this ESCRT-dependent convergence facilitates VPS4-dependent vacuolar clearance of misfolded cytoplasmic and nuclear proteins, the latter entailing extrusion of nuclear INQ into the vacuole. We propose perinuclear ESCRT coordinates spatial PQC at nuclear-vacuolar contacts to facilitate vacuolar clearance of nuclear and cytoplasmic misfolded proteins.
]]></description>
<dc:creator>Sontag, E. M.</dc:creator>
<dc:creator>Morales-Polanco, F.</dc:creator>
<dc:creator>Chen, J.-H.</dc:creator>
<dc:creator>McDermott, G.</dc:creator>
<dc:creator>Dolan, P. T.</dc:creator>
<dc:creator>Gestaut, D. R.</dc:creator>
<dc:creator>LeGros, M. A.</dc:creator>
<dc:creator>Larabell, C.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:date>2022-12-02</dc:date>
<dc:identifier>doi:10.1101/2022.12.01.518779</dc:identifier>
<dc:title><![CDATA[An ESCRT-dependent pathway of Nuclear and Cytoplasmic Spatial PQC is coordinated at Nuclear Vacuolar Junctions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.01.518665v1?rss=1">
<title>
<![CDATA[
Allosteric SHP2 Inhibition Increases Apoptotic Dependency on BCL2 and Synergizes with Venetoclax in FLT3- and KIT- Mutant AML 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.01.518665v1?rss=1</link>
<description><![CDATA[
Mutations in receptor tyrosine kinases (RTKs) FLT3 and KIT are frequent and associated with poor outcomes in acute myeloid leukemia (AML). Although FLT3 inhibitors (FLT3i) are clinically effective, remissions are short-lived due to secondary resistance characterized by acquired mutations constitutively activating the RAS/MAPK pathway. Hereby, we report pre-clinical efficacy of co-targeting SHP2, a critical node in MAPK signaling, and BCL2 in RTK-driven AML. The allosteric SHP2 inhibitor RMC-4550 suppressed proliferation of AML cell lines with FLT3 and KIT mutations, including cell lines with acquired resistance to FLT3i. We demonstrate that SHP2 inhibition unveils an Achilles heel of AML, increasing apoptotic dependency on BCL2 via MAPK-dependent mechanisms, including upregulation of BMF and downregulation of MCL1. Consequently, RMC-4550 and venetoclax are synergistically lethal in FLT3- or KIT-mutant AML cell lines, and in clinically relevant xenograft models. Our results provide new mechanistic rationale and preclinical evidence for co-targeting SHP2 and BCL2 in RTK-driven AML.

SignificanceThere is an unmet need for effective therapies targeting the MAPK pathway to overcome resistance in RTK-driven AML. We report that pharmacologic co-inhibition of SHP2 and BCL2 has synergistic anti-leukemia activity in preclinical models of AML with FLT3 and KIT mutations and holds potential clinical utility.
]]></description>
<dc:creator>Popescu, B.</dc:creator>
<dc:creator>Stahlhut, C.</dc:creator>
<dc:creator>Tarver, T. C.</dc:creator>
<dc:creator>Wishner, S.</dc:creator>
<dc:creator>Lee, B. J.</dc:creator>
<dc:creator>Peretz, C. A. C.</dc:creator>
<dc:creator>Luck, C.</dc:creator>
<dc:creator>Phojanakong, P.</dc:creator>
<dc:creator>Camara Serrano, J. A.</dc:creator>
<dc:creator>Hongo, H.</dc:creator>
<dc:creator>Rivera, J. M.</dc:creator>
<dc:creator>Xirenayi, S.</dc:creator>
<dc:creator>Chukinas, J. A.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Tasian, S. K.</dc:creator>
<dc:creator>Stieglitz, E.</dc:creator>
<dc:creator>Smith, C. C.</dc:creator>
<dc:date>2022-12-03</dc:date>
<dc:identifier>doi:10.1101/2022.12.01.518665</dc:identifier>
<dc:title><![CDATA[Allosteric SHP2 Inhibition Increases Apoptotic Dependency on BCL2 and Synergizes with Venetoclax in FLT3- and KIT- Mutant AML]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.05.518892v1?rss=1">
<title>
<![CDATA[
Generation of adult hippocampal neural stem cells occurs in the early postnatal dentate gyrus and depends on cyclin D2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.05.518892v1?rss=1</link>
<description><![CDATA[
In the hippocampus, lifelong neurogenesis is maintained by a pool of multipotent adult neural stem cells (aNSCs) residing in the subgranular zone of the dentate gyrus (DG). Yet, the mechanisms guiding the transition of NSCs from developmental to adult remain unclear. By using nestin-reporter mice deficient for D2, a cyclin expressed mainly postnatally, we show that the aNSC pool is established through D2-dependent proliferation during the first two weeks of life. The absence of D2 allows the normal development of the DG until birth but prevents the postnatal formation of radial glia-like aNSCs. Additionally, retroviral fate mapping demonstrates that aNSCs are born on-site from precursors located in the DG shortly after birth. Altogether, our data suggest that aNSCs are a population distinct from developmental NSCs and thus imply that adult hippocampal neurogenesis is not a mere continuation of development.
]]></description>
<dc:creator>Pastor-Alonso, O.</dc:creator>
<dc:creator>Syeda Zahra, A.</dc:creator>
<dc:creator>Kaske, B.</dc:creator>
<dc:creator>Garcia-Moreno, F.</dc:creator>
<dc:creator>Tetzlaff, F.</dc:creator>
<dc:creator>Bockelmann, E.</dc:creator>
<dc:creator>Grunwald, V.</dc:creator>
<dc:creator>Martin-Suarez, S.</dc:creator>
<dc:creator>Riecken, K.</dc:creator>
<dc:creator>Witte, O. W.</dc:creator>
<dc:creator>Encinas, J. M.</dc:creator>
<dc:creator>Urbach, A.</dc:creator>
<dc:date>2022-12-05</dc:date>
<dc:identifier>doi:10.1101/2022.12.05.518892</dc:identifier>
<dc:title><![CDATA[Generation of adult hippocampal neural stem cells occurs in the early postnatal dentate gyrus and depends on cyclin D2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.05.519047v1?rss=1">
<title>
<![CDATA[
Structural basis for CaVα2δ:gabapentin binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.05.519047v1?rss=1</link>
<description><![CDATA[
Gabapentinoid drugs for pain and anxiety act on the CaV2{delta}-1 and CaV2{delta}-2 subunits of high-voltage activated calcium channels (CaV1s and CaV2s). Here, we present the cryo-EM structure of the gabapentin-bound brain and cardiac CaV1.2/CaV{beta}3/CaV2{delta}-1 channel. The data reveal a binding pocket in the CaV2{delta}-1 dCache1 domain that completely encapsulates gabapentin and define CaV2{delta} isoform sequence variations that explain gabapentin binding selectivity of CaV2{delta}-1 and CaV2{delta}-2.
]]></description>
<dc:creator>Minor, D. L.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Mondal, A.</dc:creator>
<dc:date>2022-12-05</dc:date>
<dc:identifier>doi:10.1101/2022.12.05.519047</dc:identifier>
<dc:title><![CDATA[Structural basis for CaVα2δ:gabapentin binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.05.519162v1?rss=1">
<title>
<![CDATA[
Visualizing somatic alterations in spatial transcriptomics data of skin cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.05.519162v1?rss=1</link>
<description><![CDATA[
Tools to visualize genetic alterations within tissues remain underdeveloped despite the growth of spatial transcriptomic technologies, which measure gene expression in different regions of tissues. Since genetic alterations can be detected in RNA-sequencing data, we explored the feasibility of observing somatic alterations in spatial transcriptomics data. Extracting genetic information from spatial transcriptomic data would illuminate the spatial distribution of clones and allow for correlations with regional changes in gene expression to support genotype-phenotype studies. Recent work demonstrates that copy number alterations can be inferred from spatial transcriptomics data1. Here, we describe new software to further enhance the inference of copy number from spatial transcriptomics data. Moreover, we demonstrate that single nucleotide variants are also detectable in spatial transcriptomic data. We applied these approaches to map the location of point mutations, copy number alterations, and allelic imbalances in spatial transcriptomic data of two cutaneous squamous cell carcinomas. We show that both tumors are dominated by a single clone of cells, suggesting that their regional variations in gene expression2 are likely driven by non-genetic factors. Furthermore, we observe mutant cells in histologically normal tissue surrounding one tumor, which were not discernible upon histopathologic evaluation. Finally, we detected mono-allelic expression of immunoglobulin heavy chains in B-cells, revealing clonal populations of plasma cells surrounding one tumor. In summary, we put forward solutions to add the genetic dimension to spatial transcriptomic datasets, augmenting the potential of this new technology.
]]></description>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Chang, D.</dc:creator>
<dc:creator>Tandukar, B.</dc:creator>
<dc:creator>Deivendran, D.</dc:creator>
<dc:creator>Cho, R.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Bastian, B. C.</dc:creator>
<dc:creator>Ji, A. L.</dc:creator>
<dc:creator>Shain, A. H.</dc:creator>
<dc:date>2022-12-07</dc:date>
<dc:identifier>doi:10.1101/2022.12.05.519162</dc:identifier>
<dc:title><![CDATA[Visualizing somatic alterations in spatial transcriptomics data of skin cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.05.519207v1?rss=1">
<title>
<![CDATA[
RNA conformational propensities determine cellular activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.05.519207v1?rss=1</link>
<description><![CDATA[
Cellular processes are the product of interactions between biomolecules, which associate to form biologically active complexes 1. These interactions are mediated by intermolecular contacts, which if disrupted, lead to alterations in cell physiology. Nevertheless, the formation of intermolecular contacts nearly universally requires changes in the conformations of the interacting biomolecules. As a result, binding affinity and cellular activity crucially depend not only on the strength of the contacts, but also on the inherent propensities to form binding-competent conformational states2,3. Thus, conformational penalties are ubiquitous in biology and must be known in order to quantitatively model binding energetics for protein and nucleic acid interactions4,5. However, conceptual and technological limitations have hindered our ability to dissect and quantitatively measure how conformational propensities impact cellular activity. Here, we systematically altered and determined the propensities for forming the protein-bound conformation of HIV-1 TAR RNA. These propensities quantitatively predicted the binding affinities of TAR to the RNA-binding region of the Tat protein and predicted the extent of HIV-1 Tat-dependent transactivation in cells. Our results establish the role of ensemble-based conformational propensities in cellular activity and reveal an example of a cellular process driven by an exceptionally rare and short-lived RNA conformational state.
]]></description>
<dc:creator>Kelly, M. L.</dc:creator>
<dc:creator>Roy, R.</dc:creator>
<dc:creator>Geng, A.</dc:creator>
<dc:creator>Ganser, L. R.</dc:creator>
<dc:creator>Manghrani, A.</dc:creator>
<dc:creator>Cullen, B. R.</dc:creator>
<dc:creator>Schulze-Gahmen, U.</dc:creator>
<dc:creator>Herschlag, D.</dc:creator>
<dc:creator>Al-Hashimi, H. M.</dc:creator>
<dc:date>2022-12-07</dc:date>
<dc:identifier>doi:10.1101/2022.12.05.519207</dc:identifier>
<dc:title><![CDATA[RNA conformational propensities determine cellular activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.07.519375v1?rss=1">
<title>
<![CDATA[
Pleiotropy of autism-associated chromatin regulators 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.07.519375v1?rss=1</link>
<description><![CDATA[
Gene ontology analyses of high confidence autism spectrum disorder (hcASD) risk genes have historically highlighted chromatin regulation and synaptic function as major contributors to pathobiology. Our recent functional work in vivo has additionally implicated microtubule biology and identified disrupted cellular proliferation as a convergent ASD phenotype. As many chromatin regulators, including ASD risk genes ADNP and CHD3, are known to directly regulate both tubulins and histones, we studied the five chromatin regulators most strongly associated with ASD (ADNP, CHD8, CHD2, POGZ, and SUV420H1/KMT5B) specifically with respect to microtubule biology. We observe that all five localize to microtubules of the mitotic spindle in vitro and in vivo. Further in-depth investigation of CHD2 provides evidence that patient-derived mutations lead to a range of microtubule-related phenotypes, including disrupted localization of the protein at the mitotic spindle, spindle defects, cell cycle stalling, DNA damage, and cell death. Lastly, we observe that ASD genetic risk is significantly enriched among microtubule-associated proteins, suggesting broader relevance. Together, these results provide further evidence that the role of tubulin biology and cellular proliferation in ASD warrant further investigation and highlight the pitfalls of relying solely on annotated gene functions in the search for pathological mechanisms.
]]></description>
<dc:creator>Lasser, M.</dc:creator>
<dc:creator>Sun, N.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Law, K.</dc:creator>
<dc:creator>Gonzalez, S.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Drury, V.</dc:creator>
<dc:creator>Drake, S.</dc:creator>
<dc:creator>Zaltsman, Y.</dc:creator>
<dc:creator>Dea, J.</dc:creator>
<dc:creator>Bader, E.</dc:creator>
<dc:creator>McCluskey, K. E.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Willsey, A. J.</dc:creator>
<dc:creator>Willsey, H. R.</dc:creator>
<dc:date>2022-12-07</dc:date>
<dc:identifier>doi:10.1101/2022.12.07.519375</dc:identifier>
<dc:title><![CDATA[Pleiotropy of autism-associated chromatin regulators]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.07.519455v1?rss=1">
<title>
<![CDATA[
Mega-scale movie-fields in the mouse visuo-hippocampal network 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.07.519455v1?rss=1</link>
<description><![CDATA[
Natural experience often involves a continuous series of related images while the subject is immobile. How does the cortico-hippocampal circuit process this information? The hippocampus is crucial for episodic memory1-3, but most rodent single unit studies require spatial exploration4-6 or active engagement7. Hence, we investigated neural responses to a silent, isoluminant, black and white movie in head-fixed mice without any task or locomotion demands, or rewards, from the Allen Brain Observatory. The activity of most neurons (97%, 6554/6785) in the thalamo-cortical visual areas was significantly modulated by the 30s long movie clip. Surprisingly, a third (33%, 3379/10263) of hippocampal -dentate gyrus, CA1 and subiculum- neurons showed movie-selectivity, with elevated firing in specific movie sub-segments, termed movie-fields. Movie-tuning remained intact when mice were immobile or ran spontaneously. On average, a tuned cell had more than 5 movie-fields in visual areas, but only 2 in hippocampal areas. The movie-field durations in all brain regions spanned an unprecedented 1000-fold range: from 0.02s to 20s, termed mega-scale coding. Yet, the total duration of all the movie-fields of a cell was comparable across neurons and brain regions. We hypothesize that hippocampal responses show greater continuous-sequence encoding than visual areas, as evidenced by fewer and broader movie-fields than in visual areas. Consistent with this hypothesis, repeated presentation of the movie images in a fixed, scrambled sequence virtually abolished hippocampal but not visual-cortical selectivity. The enhancement of continuous movie tuning compared to the scrambled sequence was eight-fold greater in hippocampal than visual areas, further supporting episodic-sequence encoding. Thus, all mouse-brain areas investigated encoded segments of the movie. Similar results are likely to hold in primates and humans. Hence, movies could provide a unified way to probe neural mechanisms of episodic information processing and memory, even in immobile subjects, across brain regions, and species.
]]></description>
<dc:creator>Purandare, C.</dc:creator>
<dc:creator>Mehta, M.</dc:creator>
<dc:date>2022-12-07</dc:date>
<dc:identifier>doi:10.1101/2022.12.07.519455</dc:identifier>
<dc:title><![CDATA[Mega-scale movie-fields in the mouse visuo-hippocampal network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.07.519508v1?rss=1">
<title>
<![CDATA[
Mild SARS-CoV-2 infection results in long-lasting microbiota instability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.07.519508v1?rss=1</link>
<description><![CDATA[
Viruses targeting mammalian cells can indirectly alter the gut microbiota, potentially compounding their phenotypic effects. Multiple studies have observed a disrupted gut microbiota in severe cases of SARS-CoV-2 infection that require hospitalization. Yet, despite demographic shifts in disease severity resulting in a large and continuing burden of non-hospitalized infections, we still know very little about the impact of mild SARS-CoV-2 infection on the gut microbiota in the outpatient setting. To address this knowledge gap, we longitudinally sampled 14 SARS-CoV-2 positive subjects who remained outpatient and 4 household controls. SARS-CoV-2 cases exhibited a significantly less stable gut microbiota relative to controls, as long as 154 days after their positive test. These results were confirmed and extended in the K18-hACE2 mouse model, which is susceptible to SARS-CoV-2 infection. All of the tested SARS-CoV-2 variants significantly disrupted the mouse gut microbiota, including USA-WA1/2020 (the original variant detected in the United States), Delta, and Omicron. Surprisingly, despite the fact that the Omicron variant caused the least severe symptoms in mice, it destabilized the gut microbiota and led to a significant depletion in Akkermansia muciniphila. Furthermore, exposure of wild-type C57BL/6J mice to SARS-CoV-2 disrupted the gut microbiota in the absence of severe lung pathology.

IMPORTANCETaken together, our results demonstrate that even mild cases of SARS-CoV-2 can disrupt gut microbial ecology. Our findings in non-hospitalized individuals are consistent with studies of hospitalized patients, in that reproducible shifts in gut microbial taxonomic abundance in response to SARS-CoV-2 have been difficult to identify. Instead, we report a long-lasting instability in the gut microbiota. Surprisingly, our mouse experiments revealed an impact of the Omicron variant, despite producing the least severe symptoms in genetically susceptible mice, suggesting that despite the continued evolution of SARS-CoV-2 it has retained its ability to perturb the intestinal mucosa. These results will hopefully renew efforts to study the mechanisms through which Omicron and future SARS-CoV-2 variants alter gastrointestinal physiology, while also considering the potentially broad consequences of SARS-CoV-2-induced microbiota instability for host health and disease.
]]></description>
<dc:creator>Upadhyay, V.</dc:creator>
<dc:creator>Suryawanshi, R.</dc:creator>
<dc:creator>Tasoff, P.</dc:creator>
<dc:creator>McCavitt-Malvido, M.</dc:creator>
<dc:creator>Kumar, G. R.</dc:creator>
<dc:creator>Murray, V.</dc:creator>
<dc:creator>Noecker, C.</dc:creator>
<dc:creator>Bisanz, J. E.</dc:creator>
<dc:creator>Hswen, Y.</dc:creator>
<dc:creator>Ha, C.</dc:creator>
<dc:creator>Sreekumar, B.</dc:creator>
<dc:creator>Chen, I. P.</dc:creator>
<dc:creator>Lynch, S. V.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:date>2022-12-08</dc:date>
<dc:identifier>doi:10.1101/2022.12.07.519508</dc:identifier>
<dc:title><![CDATA[Mild SARS-CoV-2 infection results in long-lasting microbiota instability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.06.519274v1?rss=1">
<title>
<![CDATA[
Novel Peptide Inhibitor of Human Tumor Necrosis Factor-α has Antiarthritic Activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.06.519274v1?rss=1</link>
<description><![CDATA[
The inhibition of tumor necrosis factor- (TNF) trimer formation renders it inactive for binding to its receptors thus mitigating the vicious cycle of inflammation. We designed a peptide (PIYLGGVFQ) that simulates a sequence strand of human TNF monomer using a series of in silico methods, such as active site finding (Acsite), protein-protein interaction (PPI), docking studies (GOLD and Modeller) followed by molecular dynamics (MD) simulation studies. The MD studies confirmed the intermolecular interaction of the peptide with the TNF. Fluorescence-activated cell sorting (FACS) and fluorescence microscopy revealed that the peptide effectively inhibited the binding of TNF to the cell surface receptors. The cell culture assays showed that the peptide significantly inhibited the TNF-mediated cell death. In addition, the nuclear translocation of the nuclear factor kappa B (NF{kappa}B) was significantly suppressed in the peptide-treated A549 cells as observed in immunofluorescence and gel mobility-shift assays. Furthermore, peptide protected against joint damage in collagen-induced arthritis (CIA) mouse model as revealed in the microfocal-CT scans. In conclusion, this TNF antagonist would be useful for the prevention and repair of inflammatory bone destruction and subsequent loss in the mouse model of CIA as well as human rheumatoid arthritis (RA) patients. This calls upon further clinical investigation to utilize its potential effect as an anti-arthritic drug.
]]></description>
<dc:creator>Sahu, D.</dc:creator>
<dc:creator>Gupta, C.</dc:creator>
<dc:creator>Yennamalli, R.</dc:creator>
<dc:creator>Sharma, S.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:creator>Hasan, S.</dc:creator>
<dc:creator>Gupta, P.</dc:creator>
<dc:creator>Sharma, V. K.</dc:creator>
<dc:creator>Kashyap, S.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Dwivedi, V. P.</dc:creator>
<dc:creator>Panda, A. K.</dc:creator>
<dc:creator>Das, H. R.</dc:creator>
<dc:creator>Liu, C.-J.</dc:creator>
<dc:date>2022-12-10</dc:date>
<dc:identifier>doi:10.1101/2022.12.06.519274</dc:identifier>
<dc:title><![CDATA[Novel Peptide Inhibitor of Human Tumor Necrosis Factor-α has Antiarthritic Activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.07.519545v1?rss=1">
<title>
<![CDATA[
Increased mRNA expression of CDKN2A is a transcriptomic marker of clinically aggressive meningiomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.07.519545v1?rss=1</link>
<description><![CDATA[
BackgroundHomozygous loss of CDKN2A/B is a genetic alteration found in many cancer types including meningiomas, where it is associated with poor clinical outcome. It is now also a diagnostic criterion for grade 3 meningiomas in the 2021 WHO classification for central nervous system tumors. However, as in other cancers, the relationship between copy number loss of CDKN2A/B and expression of its gene product is unclear and may be either commensurate or paradoxical in nature. Therefore, we aimed to investigate the association of CDKN2A mRNA expression with clinical prognosis, WHO grade, and other molecular biomarkers in meningiomas such as DNA methylation, molecular group, and proteomics.

MethodsWe used multidimensional molecular data of 490 meningioma samples from 4 independent cohorts to examine the relationship between mRNA expression of CDKN2A and copy number status, its correlation to clinical outcome, the transcriptomic pathways altered in differential CDKN2A expression, and its relationship with DNA methylation, and proteomics using an integrated molecular approach.

ResultsMeningiomas without any copy number loss were dichotomized into high (CDKN2Ahigh) and low (CDKN2Alow) CDKN2A mRNA expression groups. Patients with CDKN2Ahigh meningiomas had poorer progression free survival (PFS) compared to those with CDKN2Alow meningiomas. CDKN2A mRNA expression was increased in more aggressive molecular groups, and in higher WHO grade meningiomas across all cohorts. CDKN2Ahigh meningiomas and meningiomas with CDKN2A copy number loss shared common up-regulated cell cycling pathways. CDK4 mRNA expression was increased in CDKN2Ahigh meningiomas and both p16 and CDK4 protein were more abundant in CDKN2Ahigh meningiomas. CDKN2Ahigh meningiomas were frequently hypermethylated at the gene body and UTR compared to CDKN2Alow meningiomas and found be more commonly Rb-deficient.

ConclusionsAn intermediate level of CDKN2A mRNA expression appears to be optimal as significantly low (CDKN2A deleted) or high expression (CDKN2Ahigh) are associated with poorer outcomes clinically. Though CDK4 is elevated in CDKN2Ahigh meningiomas, Rb-deficiency may be more common in this group, leading to lack of response to CDK inhibitors.
]]></description>
<dc:creator>Wang, J. Z.</dc:creator>
<dc:creator>Patil, V.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Dogan, H.</dc:creator>
<dc:creator>Behling, F.</dc:creator>
<dc:creator>Skardelly, M.</dc:creator>
<dc:creator>Tatagiba, M.</dc:creator>
<dc:creator>Hoffman, E.</dc:creator>
<dc:creator>Bunda, S.</dc:creator>
<dc:creator>Yakubov, R.</dc:creator>
<dc:creator>Kaloti, R.</dc:creator>
<dc:creator>Brandner, S.</dc:creator>
<dc:creator>Gao, A.</dc:creator>
<dc:creator>Cohen-Gadol, A.</dc:creator>
<dc:creator>Barnholtz-Sloan, J.</dc:creator>
<dc:creator>Raleigh, D.</dc:creator>
<dc:creator>Sahm, F.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:creator>Tabatabai, G.</dc:creator>
<dc:creator>Aldape, K.</dc:creator>
<dc:creator>Nassiri, F.</dc:creator>
<dc:creator>Zadeh, G.</dc:creator>
<dc:creator>International Consortium on Meningiomas (ICOM),</dc:creator>
<dc:date>2022-12-10</dc:date>
<dc:identifier>doi:10.1101/2022.12.07.519545</dc:identifier>
<dc:title><![CDATA[Increased mRNA expression of CDKN2A is a transcriptomic marker of clinically aggressive meningiomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.10.519926v1?rss=1">
<title>
<![CDATA[
Bulk and Mosaic Deletions of Egfr Reveal Regionally Defined Gliogenesis in the Developing Mouse Forebrain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.10.519926v1?rss=1</link>
<description><![CDATA[
The Epidermal growth factor receptor (EGFR) plays a role in cell proliferation and differentiation during healthy development and tumor growth, however its requirement for brain development remains unclear. Here we used a conditional mouse allele for Egfr to examine its contributions to perinatal forebrain development at the tissue level. Subtractive bulk ventral and dorsal forebrain deletions of Egfr uncovered significant and permanent decreases in oligodendrogenesis and myelination in the cortex and corpus callosum. Additionally, an increase in astrogenesis or reactive astrocytes in effected regions was evident in response to cortical scarring. Sparse deletion using Mosaic Analysis with Double Markers (MADM) surprisingly revealed a regional requirement for EGFR in rostrodorsal, but not ventrocaudal glial lineages including both astrocytes and oligodendrocytes. The EGFR-independent ventral glial progenitors may compensate for the missing EGFR-dependent dorsal glia in the bulk Egfr-deleted forebrain, potentially exposing a regenerative population of gliogenic progenitors in the mouse forebrain.
]]></description>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Xiao, G.</dc:creator>
<dc:creator>Johnson, C. A.</dc:creator>
<dc:creator>Cai, Y.</dc:creator>
<dc:creator>Horowitz, Z.</dc:creator>
<dc:creator>Mennicke, C.</dc:creator>
<dc:creator>Coffey, R.</dc:creator>
<dc:creator>Haider, M.</dc:creator>
<dc:creator>Threadgill, D. W.</dc:creator>
<dc:creator>Eliscu, R.</dc:creator>
<dc:creator>Oldham, M. C.</dc:creator>
<dc:creator>Greenbaum, A. C.</dc:creator>
<dc:creator>Ghashghaei, H. T.</dc:creator>
<dc:date>2022-12-11</dc:date>
<dc:identifier>doi:10.1101/2022.12.10.519926</dc:identifier>
<dc:title><![CDATA[Bulk and Mosaic Deletions of Egfr Reveal Regionally Defined Gliogenesis in the Developing Mouse Forebrain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.09.519807v1?rss=1">
<title>
<![CDATA[
3D multiplexed tissue imaging reconstruction and optimized region-of-interest (ROI) selection through deep learning model of channels embedding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.09.519807v1?rss=1</link>
<description><![CDATA[
Tissue-based sampling and diagnosis are defined as the extraction of information from certain limited spaces and its diagnostic significance of a certain object. Pathologists deal with issues related to tumor heterogeneity since analyzing a single sample does not necessarily capture a representative depiction of cancer, and a tissue biopsy usually only presents a small fraction of the tumor. Many multiplex tissue imaging platforms (MTIs) make the assumption that tissue microarrays (TMAs) containing small core samples of 2-dimensional (2D) tissue sections are a good approximation of bulk tumors although tumors are not 2D. However, emerging whole slide imaging (WSI) or 3D tumor atlases that employ MTIs like cyclic immunofluorescence (CyCIF) strongly challenge this assumption. In spite of the additional insight gathered by measuring the tumor microenvironment in WSI or 3D, it can be prohibitively expensive and time-consuming to process tens or hundreds of tissue sections with CyCIF. Even when resources are not limited, the criteria for region-of-interest (ROI) selection in tissues for downstream analysis remain largely qualitative and subjective as stratified sampling requires the knowledge of objects and evaluates their features. Despite the fact TMAs fail to adequately approximate whole tissue features, a theoretical subsampling of tissue exists that can best represent the tumor in the whole slide image. To address these challenges, we propose deep learning approaches to learn multi-modal image translation tasks from two aspects: 1) generative modeling approach to reconstruct 3D CyCIF representation and 2) co-embedding CyCIF image and Hematoxylin and Eosin (H&E) section to learn multi-modal mappings by a cross-domain translation for minimum representative ROI selection. We demonstrate that generative modeling enables a 3D virtual CyCIF reconstruction of a colorectal cancer specimen given a small subset of the imaging data at training time. By co-embedding histology and MTI features, we propose a simple convex optimization for objective ROI selection. We demonstrate the potential application of ROI selection and the efficiency of its performance with respect to cellular heterogeneity.
]]></description>
<dc:creator>Burlingame, E.</dc:creator>
<dc:creator>Ternes, L.</dc:creator>
<dc:creator>Lin, J.-R.</dc:creator>
<dc:creator>Chen, Y.-A.</dc:creator>
<dc:creator>Kim, E. N.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Santagata, S.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2022-12-12</dc:date>
<dc:identifier>doi:10.1101/2022.12.09.519807</dc:identifier>
<dc:title><![CDATA[3D multiplexed tissue imaging reconstruction and optimized region-of-interest (ROI) selection through deep learning model of channels embedding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.13.520194v1?rss=1">
<title>
<![CDATA[
Neural circuit-wide analysis of gene expression during deafening-induced destabilization ofbirdsong 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.13.520194v1?rss=1</link>
<description><![CDATA[
Sensory feedback is required for stable execution of learned motor skills, and its loss can severely disrupt motor performance. The neural mechanisms that mediate sensorimotor stability have been extensively studied at systems and physiological levels, yet relatively little is known about how disruptions to sensory input alter the molecular properties of associated motor systems. Songbird courtship song, a model for skilled behavior, is a learned and highly structured vocalization that is destabilized following deafening. Here, we sought to determine how the loss of auditory feedback modifies gene expression and its coordination across the birdsong sensorimotor circuit. To facilitate this system-wide analysis of transcriptional responses, we developed a gene expression profiling approach that enables the construction of hundreds of spatially-defined RNA-sequencing libraries. Using this method, we found that deafening preferentially alters gene expression across birdsong neural circuitry relative to surrounding areas, particularly in premotor and striatal regions. Genes with altered expression are associated with synaptic transmission, neuronal spines, and neuromodulation and show a bias toward expression in glutamatergic neurons and Pvalb/Sst-class GABAergic interneurons. We also found that connected song regions exhibit correlations in gene expression that were reduced in deafened birds relative to hearing birds, suggesting that song destabilization alters the inter-region coordination of transcriptional state. Finally, lesioning LMAN, a forebrain afferent of RA required for deafening-induced song plasticity, had the largest effect on groups of genes that were also most affected by deafening. Combined, this integrated transcriptomics analysis demonstrates that the loss of peripheral sensory input drives a distributed gene expression response throughout associated sensorimotor neural circuitry and identifies specific candidate molecular and cellular mechanisms that support stability and plasticity of learned motor skills.
]]></description>
<dc:creator>Colquitt, B. M.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Green, F.</dc:creator>
<dc:creator>Veline, R. J.</dc:creator>
<dc:creator>Brainard, M. S.</dc:creator>
<dc:date>2022-12-14</dc:date>
<dc:identifier>doi:10.1101/2022.12.13.520194</dc:identifier>
<dc:title><![CDATA[Neural circuit-wide analysis of gene expression during deafening-induced destabilization ofbirdsong]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.14.518457v1?rss=1">
<title>
<![CDATA[
A multiomics approach reveals RNA dynamics promote cellular sensitivity to DNA hypomethylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.14.518457v1?rss=1</link>
<description><![CDATA[
The search for new approaches in cancer therapy requires a mechanistic understanding of cancer vulnerabilities and anti-cancer drug mechanisms of action. Problematically, some effective therapeutics target cancer vulnerabilities that have poorly defined mechanisms of anti-cancer activity. One such drug is decitabine, a frontline therapeutic approved for the treatment of high-risk acute myeloid leukemia (AML). Decitabine is thought to kill cancer cells selectively via inhibition of DNA methyltransferase enzymes, but the genes and mechanisms involved remain unclear. Here, we apply an integrated multiomics and CRISPR functional genomics approach to identify genes and processes associated with response to decitabine in AML cells. Our integrated multiomics approach reveals RNA dynamics are key regulators of DNA hypomethylation induced cell death. Specifically, regulation of RNA decapping, splicing and RNA methylation emerge as important regulators of cellular response to decitabine.
]]></description>
<dc:creator>Ge, A. Y.</dc:creator>
<dc:creator>Arab, A.</dc:creator>
<dc:creator>Dai, R.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Fish, L.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Asgharian, H.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Gilbert, L.</dc:creator>
<dc:date>2022-12-14</dc:date>
<dc:identifier>doi:10.1101/2022.12.14.518457</dc:identifier>
<dc:title><![CDATA[A multiomics approach reveals RNA dynamics promote cellular sensitivity to DNA hypomethylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.14.520438v1?rss=1">
<title>
<![CDATA[
Effects of DNA extraction, DNA integrity, and laboratory on the precision of qPCR-based telomere length measurement - a multi-lab impartial study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.14.520438v1?rss=1</link>
<description><![CDATA[
Measuring telomere length (TL) with high precision is challenging. Currently there is insufficient understanding of the causes of variation in measurement precision, particularly for qPCR-based measurement. To better understand how DNA extraction protocols and laboratory-specific analytical factors influence qPCR-based TL measurement precision, we conducted a multi-laboratory study involving four labs and six DNA extraction protocols assaying the same blinded human whole blood samples. DNA extraction protocols differed in underlying principles (magnetic beads, salting out, silica membrane) and commercial kits. A fifth lab performed Telomere Restriction Fragment (TRF) analysis using Southern Blot technique with one DNA extraction protocol. All labs performed TL measurement using their standard procedures on two sets of fifty double blinded samples. Data was sent to a central point for unblinding and statistical analyses. Precision was quantified using the Intraclass Correlation Coefficient (ICC). Correlations with TRF measurements were also calculated. Repeated qPCR-based measurements of the same DNA extraction yielded ICC values ranging from 0.24 to 0.94. ICC values calculated over measurements of repeated DNA extractions were on average 0.23 lower and ranged from 0.02 to 0.83. The latter ICC estimates more strongly predicted the association between qPCR- and Southern blot-based measurements across the protocol / lab combinations (R2=0.56 vs. R2=0.93). We conclude that ICC calculated over measurements on repeated DNA extractions reliably indicates measurement precision, while ICC calculated over multiple measurements of the same DNA extraction overestimates measurement precision. Variation in ICC was driven by variation between laboratories, with few consistent DNA extraction protocol effects. Values of DNA integrity and purity generally characterized as reflecting high sample quality, (e.g. OD 260/280 of 1.8 and OD 260/230 of 2.0) were associated with qPCR-based measurement precision, but did not always predict higher ICCs.
]]></description>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Verhulst, S.</dc:creator>
<dc:creator>Fernandez Alonso, C.</dc:creator>
<dc:creator>Dagnall, C.</dc:creator>
<dc:creator>Gadalla, S.</dc:creator>
<dc:creator>Hastings, W.</dc:creator>
<dc:creator>Lai, T.-P.</dc:creator>
<dc:creator>Zheng, Y.-L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Shalev, I.</dc:creator>
<dc:creator>Epel, E.</dc:creator>
<dc:creator>Drury, S.</dc:creator>
<dc:date>2022-12-14</dc:date>
<dc:identifier>doi:10.1101/2022.12.14.520438</dc:identifier>
<dc:title><![CDATA[Effects of DNA extraction, DNA integrity, and laboratory on the precision of qPCR-based telomere length measurement - a multi-lab impartial study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.13.520297v1?rss=1">
<title>
<![CDATA[
Functional network organization of the mouse cortex determined by wide-field fluorescence imaging shares some --but not all-- properties revealed with simultaneous fMRI-BOLD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.13.520297v1?rss=1</link>
<description><![CDATA[
Work in humans and animals shows that the brain can be decomposed into large-scale functional networks. Whereas most studies, especially in humans, use the blood-oxygenation-level-dependent (BOLD) signal, the relationship between BOLD and neuronal activity is complex and incompletely understood. This limits our ability to interpret and apply measures derived from fMRI-BOLD. Here, we employ wide-field Ca2+ imaging simultaneously recorded with fMRI-BOLD in highly-sampled mice expressing GCaMP6f in excitatory neurons. These unique data enabled us to characterize the similarities and differences between networks discoverable by each modality. Importantly, we applied a network partitioning approach that uses a mixed-membership algorithm, which allows brain regions to participate in multiple networks with varying strengths. This contrasts with assuming regions belong to only one network. Our findings demonstrate that (1) most BOLD networks are detected via Ca2+ signals. (2) There is considerable overlapping--as opposed to disjoint--network organization that is evident from both modalities. (3) Large-scale networks determined by Ca2+ signals at low temporal frequencies (0.01 - 0.5 Hz)--as opposed to higher frequencies (0.5 - 5 Hz)--are more similar to those determined by BOLD. (4) Despite many similarities, differences emerge across modes including the spatial distribution of membership diversity (the extent to which regions affiliate with multiple networks). In sum, Ca2+ imaging of excitatory neurons confirms that the mouse cortex is functionally organized into overlapping large-scale networks in a manner that reflects many, but not all, properties observable with simultaneous fMRI-BOLD; affirming the neural origins of patterns of brain organization that are evident in a clinically accessible neuroimaging modality.
]]></description>
<dc:creator>Vafaii, H.</dc:creator>
<dc:creator>Mandino, F.</dc:creator>
<dc:creator>Desrosiers-Gregoire, G.</dc:creator>
<dc:creator>O'Connor, D.</dc:creator>
<dc:creator>Shen, X.</dc:creator>
<dc:creator>Ge, X.</dc:creator>
<dc:creator>Herman, P.</dc:creator>
<dc:creator>Hyder, F.</dc:creator>
<dc:creator>Papademetris, X.</dc:creator>
<dc:creator>Chakravarty, M.</dc:creator>
<dc:creator>Crair, M.</dc:creator>
<dc:creator>Constable, R. T.</dc:creator>
<dc:creator>Lake, E. M.</dc:creator>
<dc:creator>Pessoa, L.</dc:creator>
<dc:date>2022-12-15</dc:date>
<dc:identifier>doi:10.1101/2022.12.13.520297</dc:identifier>
<dc:title><![CDATA[Functional network organization of the mouse cortex determined by wide-field fluorescence imaging shares some --but not all-- properties revealed with simultaneous fMRI-BOLD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.15.520648v1?rss=1">
<title>
<![CDATA[
The FMRF-NH2 Gated Sodium Channel of Biomphalaria glabrata: Localization and Expression Following Infection by Schistosoma mansoni 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.15.520648v1?rss=1</link>
<description><![CDATA[
The neglected tropical disease schistosomiasis impacts the lives of over 700 million people globally. Schistosoma mansoni, the trematode parasite that causes the most common type of schistosomiasis, requires planorbid pond snails of the genus Biomphalaria to support its larval development and transformation to the form that can infect humans. A greater understanding of neural signaling systems that are specific to the Biomphalaria intermediate host could lead to novel strategies for parasite or snail control. This study characterized a Biomphalaria glabrata neural receptor that is gated by the molluscan neuropeptide FMRF-NH2. The Biomphalaria glabrata FMRF-NH2 gated sodium channel (Bgl-FaNaC) amino acid sequence was highly conserved with FaNaCs found in related gastropods, especially the planorbid Planorbella trivolvis (91% sequence identity). In common with the P. trivolvis FaNaC, the B. glabrata receptor exhibited a low affinity (EC50: 3 x 10-4 M) and high specificity for the FMRF-NH2 agonist. Its expression in the central nervous system, detected by immunohistochemistry and in situ hybridization, was widespread, with the protein localized mainly to neuronal fibers and the mRNA confined to cell bodies. Colocalization was observed with the FMRF-NH2 tetrapeptide precursor in some neurons associated with male mating behavior. At the mRNA level, Bgl-FaNaC expression in the visceral and left parietal ganglia decreased at 20 days post infection by S. mansoni and in the shedding phase. Altered FMRF-NH2 signaling could be vital for parasite survival and proliferation in its snail intermediate host.
]]></description>
<dc:creator>Miller, M. W.</dc:creator>
<dc:creator>Vicente, L. C.</dc:creator>
<dc:creator>Behra, M.</dc:creator>
<dc:creator>Torres, A.</dc:creator>
<dc:creator>Mendez, P.</dc:creator>
<dc:creator>Rosa-Casillas, M.</dc:creator>
<dc:creator>Bracho, D. P.</dc:creator>
<dc:creator>Rosenthal, J. J.</dc:creator>
<dc:date>2022-12-15</dc:date>
<dc:identifier>doi:10.1101/2022.12.15.520648</dc:identifier>
<dc:title><![CDATA[The FMRF-NH2 Gated Sodium Channel of Biomphalaria glabrata: Localization and Expression Following Infection by Schistosoma mansoni]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.16.520768v1?rss=1">
<title>
<![CDATA[
Direct neural perturbations reveal a dynamical mechanism for robust computation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.16.520768v1?rss=1</link>
<description><![CDATA[
The rich repertoire of skilled mammalian behavior is the product of neural circuits that generate robust and flexible patterns of activity distributed across populations of neurons. Decades of associative studies have linked many behaviors to specific patterns of population activity, but association alone cannot reveal the dynamical mechanisms that shape those patterns. Are local neural circuits high-dimensional dynamical reservoirs able to generate arbitrary superpositions of patterns with appropriate excitation? Or might circuit dynamics be shaped in response to behavioral context so as to generate only the low-dimensional patterns needed for the task at hand? Here, we address these questions within primate motor cortex by delivering optogenetic and electrical microstimulation perturbations during reaching behavior. We develop a novel analytic approach that relates measured activity to theoretically tractable, dynamical models of excitatory and inhibitory neurons. This computational model captures the dynamical effects of these perturbations and demonstrates that motor cortical activity during reaching is shaped by a self-contained, low-dimensional dynamical system. The subspace containing task-relevant dynamics proves to be oriented so as to be robust to strong non-normal amplification within cortical circuits. This task dynamics space exhibits a privileged causal relationship with behavior, in that stimulation in motor cortex perturb reach kinematics only to the extent that it alters neural states within this subspace. Our results resolve long-standing questions about the dynamical structure of cortical activity associated with movement, and illuminate the dynamical perturbation experiments needed to understand how neural circuits throughout the brain generate complex behavior.
]]></description>
<dc:creator>O'Shea, D. J.</dc:creator>
<dc:creator>Duncker, L.</dc:creator>
<dc:creator>Goo, W.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Vyas, S.</dc:creator>
<dc:creator>Trautmann, E. M.</dc:creator>
<dc:creator>Diester, I.</dc:creator>
<dc:creator>Ramakrishnan, C.</dc:creator>
<dc:creator>Deisseroth, K.</dc:creator>
<dc:creator>Sahani, M.</dc:creator>
<dc:creator>Shenoy, K. V.</dc:creator>
<dc:date>2022-12-16</dc:date>
<dc:identifier>doi:10.1101/2022.12.16.520768</dc:identifier>
<dc:title><![CDATA[Direct neural perturbations reveal a dynamical mechanism for robust computation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.19.521083v1?rss=1">
<title>
<![CDATA[
Silencing neuroinflammatory reactive astrocyte activating factors ameliorates disease outcomes in perinatal white matter injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.19.521083v1?rss=1</link>
<description><![CDATA[
The role of reactive astrocytes in perinatal white matter injury (WMI) is unclear. In a mouse model of WMI, we provide evidence that impairing the formation of a C3-expressing neuroinflammatory reactive astrocyte sub-state rescues myelination and behavioral deficits. We further demonstrate the presence of C3-expressing reactive astrocytes in human WMI. Our data point to these cells as putative drivers of myelination failure in WMI and a potentially promising therapeutic target.
]]></description>
<dc:creator>Renz, P. V.</dc:creator>
<dc:creator>Surbek, D.</dc:creator>
<dc:creator>Haesler, V.</dc:creator>
<dc:creator>Tscherrig, V.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Chavali, M.</dc:creator>
<dc:creator>Liddelow, S. A.</dc:creator>
<dc:creator>Rowitch, D.</dc:creator>
<dc:creator>Schoeberlein, A.</dc:creator>
<dc:creator>Brosius Lutz, A. R.</dc:creator>
<dc:date>2022-12-20</dc:date>
<dc:identifier>doi:10.1101/2022.12.19.521083</dc:identifier>
<dc:title><![CDATA[Silencing neuroinflammatory reactive astrocyte activating factors ameliorates disease outcomes in perinatal white matter injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.20.520951v1?rss=1">
<title>
<![CDATA[
Structural basis of odorant recognition by a human odorant receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.20.520951v1?rss=1</link>
<description><![CDATA[
Our sense of smell enables us to navigate a vast space of chemically diverse odor molecules. This task is accomplished by the combinatorial activation of approximately 400 olfactory G protein-coupled receptors (GPCRs) encoded in the human genome1-3. How odorants are recognized by olfactory receptors (ORs) remains mysterious. Here we provide mechanistic insight into how an odorant binds a human olfactory receptor. Using cryogenic electron microscopy (cryo-EM), we determined the structure of active human OR51E2 bound to the fatty acid propionate. Propionate is bound within an occluded pocket in OR51E2 and makes specific contacts critical to receptor activation. Mutation of the odorant binding pocket in OR51E2 alters the recognition spectrum for fatty acids of varying chain length, suggesting that odorant selectivity is controlled by tight packing interactions between an odorant and an olfactory receptor. Molecular dynamics simulations demonstrate propionate-induced conformational changes in extracellular loop 3 to activate OR51E2. Together, our studies provide a high-resolution view of chemical recognition of an odorant by a vertebrate OR, providing insight into how this large family of GPCRs enables our olfactory sense.
]]></description>
<dc:creator>Billesboelle, C. B.</dc:creator>
<dc:creator>de March, C. A.</dc:creator>
<dc:creator>van der Velden, W. J. C.</dc:creator>
<dc:creator>Ma, N.</dc:creator>
<dc:creator>Tewari, J.</dc:creator>
<dc:creator>Linas del Torrent, C.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Faust, B.</dc:creator>
<dc:creator>Vaidehi, N.</dc:creator>
<dc:creator>Matsunami, H.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:date>2022-12-20</dc:date>
<dc:identifier>doi:10.1101/2022.12.20.520951</dc:identifier>
<dc:title><![CDATA[Structural basis of odorant recognition by a human odorant receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.21.521379v1?rss=1">
<title>
<![CDATA[
O-GlcNAc glycosylation orchestrates fate decision and niche function of bone marrow stromal progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.21.521379v1?rss=1</link>
<description><![CDATA[
In mammals, interactions between the bone marrow (BM) stroma and hematopoietic progenitors contribute to bone-BM homeostasis. Perinatal bone growth and ossification provide a microenvironment for the transition to definitive hematopoiesis; however, mechanisms and interactions orchestrating the development of skeletal and hematopoietic systems remain largely unknown. Here, we establish intracellular O-linked {beta}-N-acetylglucosamine (O-GlcNAc) modification as a posttranslational switch that dictates the differentiation fate and niche function of early BM stromal cells (BMSCs). By modifying and activating RUNX2, O-GlcNAcylation promotes osteogenic differentiation of BMSCs and stromal IL-7 expression to support lymphopoiesis. In contrast, C/EBP{beta}-dependent marrow adipogenesis and expression of myelopoietic stem cell factor (SCF) is inhibited by O-GlcNAcylation. Ablating O-GlcNAc transferase (OGT) in BMSCs leads to impaired bone formation, increased marrow adiposity, as well as defective B-cell lymphopoiesis and myeloid overproduction in mice. Thus, the balance of osteogenic and adipogenic differentiation of BMSCs is determined by reciprocal O-GlcNAc regulation of transcription factors, which simultaneously shapes the hematopoietic niche.
]]></description>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Huang, Z.</dc:creator>
<dc:creator>Awad, M.</dc:creator>
<dc:creator>Elsalanty, M.</dc:creator>
<dc:creator>Cray, J.</dc:creator>
<dc:creator>Ball, L. E.</dc:creator>
<dc:creator>Maynard, J.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Zeng, H.</dc:creator>
<dc:creator>Mansky, K. C.</dc:creator>
<dc:creator>Ruan, H.-B.</dc:creator>
<dc:date>2022-12-21</dc:date>
<dc:identifier>doi:10.1101/2022.12.21.521379</dc:identifier>
<dc:title><![CDATA[O-GlcNAc glycosylation orchestrates fate decision and niche function of bone marrow stromal progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.21.520137v1?rss=1">
<title>
<![CDATA[
Multi-center integrated analysis of non-coding CRISPR screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.21.520137v1?rss=1</link>
<description><![CDATA[
The ENCODE Consortiums efforts to annotate non-coding, cis-regulatory elements (CREs) have advanced our understanding of gene regulatory landscapes which play a major role in health and disease. Pooled, non-coding CRISPR screens are a promising approach for systematically investigating gene regulatory mechanisms. Here, the ENCODE Functional Characterization Centers report 109 screens comprising 346,970 individual perturbations across 13.3Mb of the genome, using a variety of methods, readouts, and statistical analyses. Across 332 functionally confirmed CRE-gene links, we identify principles for screening endogenous, non-coding elements for causal regulatory mechanisms. Nearly all CREs show strong evidence of open chromatin, and targeting accessibility peak summits is a critical component of our proposed sgRNA design rules. We provide experimental guidelines to accurately detect CREs with variable, often low, transcriptional effects. We discover a previously undescribed DNA strand-bias for CRISPRi in transcribed regions with implications for screen design and analysis. Benchmarking five screen analysis tools, we find CASA produces the most conservative CRE calls and is robust to artifacts of low-specificity sgRNAs. Together, we provide an accessible data resource, predesigned sgRNAs targeting 3,275,697 ENCODE SCREEN candidate CREs, and screening guidelines to accelerate functional characterization of the non-coding genome.
]]></description>
<dc:creator>Yao, D.</dc:creator>
<dc:creator>Tycko, J.</dc:creator>
<dc:creator>Oh, W.</dc:creator>
<dc:creator>Bounds, L. R.</dc:creator>
<dc:creator>Gosai, S. J.</dc:creator>
<dc:creator>Lataniotis, L.</dc:creator>
<dc:creator>Mackay-Smith, A.</dc:creator>
<dc:creator>Doughty, B. R.</dc:creator>
<dc:creator>Gabdank, I.</dc:creator>
<dc:creator>Schmidt, H.</dc:creator>
<dc:creator>Youngworth, I.</dc:creator>
<dc:creator>Andreeva, K.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Barrera, A.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Siklenka, K.</dc:creator>
<dc:creator>Yardimci, G. G.</dc:creator>
<dc:creator>The ENCODE4 Consortium,</dc:creator>
<dc:creator>Tewhey, R.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:creator>Greenleaf, W. J.</dc:creator>
<dc:creator>Sabeti, P. C.</dc:creator>
<dc:creator>Leslie, C.</dc:creator>
<dc:creator>Pritykin, Y.</dc:creator>
<dc:creator>Moore, J. E.</dc:creator>
<dc:creator>Beer, M. A.</dc:creator>
<dc:creator>Gersbach, C.</dc:creator>
<dc:creator>Reddy, T. E.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Engreitz, J. M.</dc:creator>
<dc:creator>Bassik, M. C.</dc:creator>
<dc:creator>Reilly, S. K.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.21.520137</dc:identifier>
<dc:title><![CDATA[Multi-center integrated analysis of non-coding CRISPR screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.22.521453v1?rss=1">
<title>
<![CDATA[
A central helical fulcrum in eIF2B coordinates allosteric regulation of Integrated Stress Response signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.22.521453v1?rss=1</link>
<description><![CDATA[
The Integrated Stress Response (ISR) enables cells to survive a variety of acute stresses, but chronic activation of the ISR underlies age-related diseases. ISR signaling down-regulates translation and activates expression of stress-responsive factors that promote return to homeostasis, and is initiated by inhibition of the decameric guanine nucleotide exchange factor eIF2B. Conformational and assembly transitions regulate eIF2B activity, but the allosteric mechanisms controlling these dynamic transitions are unknown. Using hydrogen deuterium exchange-mass spectrometry and cryo-EM, we identified a single alpha-helix whose orientation allosterically controls eIF2B conformation and assembly. Biochemical and signaling assays show that this "Switch-Helix" controls eIF2B activity and signaling in cells. In sum, the Switch-Helix acts as a fulcrum of eIF2B conformational regulation and is a highly conserved actuator of ISR signal transduction. This work uncovers a novel allosteric mechanism and unlocks new therapeutic possibilities for ISR-linked diseases.
]]></description>
<dc:creator>Lawrence, R.</dc:creator>
<dc:creator>Shoemaker, S.</dc:creator>
<dc:creator>Deal, A.</dc:creator>
<dc:creator>Sangwan, S.</dc:creator>
<dc:creator>Anand, A.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Marqusee, S.</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521453</dc:identifier>
<dc:title><![CDATA[A central helical fulcrum in eIF2B coordinates allosteric regulation of Integrated Stress Response signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.22.521501v1?rss=1">
<title>
<![CDATA[
Structural Basis of Prostaglandin Efflux by MRP4 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.22.521501v1?rss=1</link>
<description><![CDATA[
MRP4 is unique among the C family of ATP-binding cassette transporters for its role in translocating prostanoids, an important group of signaling molecules derived from unsaturated fatty acids. Using a reconstituted system, we report that a pair of prostaglandins (PGs) and the sulfonated-sterol DHEA-S preferentially enhance the ATPase activity of MRP4 over other previously proposed physiological substrates such as cyclic nucleotides or leukotrienes. We determined the cryo-EM structures of nanodisc embedded bovine MRP4 in (i) a nucleotide- and substrate-free state, (ii) in complex with PGE1, (iii) PGE2, and (iv) DHEA-S, and (v) a catalytically dead mutant E1202Q bound to ATP-Mg2+. The substrate-bound structures suggest unique features of the MRP4 binding site that distinguish its specificity for prostanoids from that of the related leukotriene transporter MRP1. The ATP-bound structure is in an outward-occluded conformation, revealing a novel state in the proposed alternate-access mechanism of MRP transport. Our study provides insights into the endogenous function of this versatile efflux transporter.
]]></description>
<dc:creator>Pourmal, S.</dc:creator>
<dc:creator>Green, E.</dc:creator>
<dc:creator>Bajaj, R.</dc:creator>
<dc:creator>Chemmama, I.</dc:creator>
<dc:creator>Knudsen, G.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Craik, C.</dc:creator>
<dc:creator>Kroetz, D.</dc:creator>
<dc:creator>Stroud, R.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521501</dc:identifier>
<dc:title><![CDATA[Structural Basis of Prostaglandin Efflux by MRP4]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.22.521524v1?rss=1">
<title>
<![CDATA[
Optimized CRISPR guide RNA library cloning reduces skew and enables more compact genetic screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.22.521524v1?rss=1</link>
<description><![CDATA[
The development of CRISPR genetic screening tools has improved functional genomics, as these tools enable precise genomic editing, provide broad access to genomic regions beyond protein-coding genes, and have fewer off-target effects than other functional genomics modalities, allowing for novel applications with smaller library sizes compared to prior technologies. Pooled functional genomics screens require high cellular coverage per perturbation to accurately quantify phenotypes and average out phenotype-independent variability across the population. While more compact libraries have decreased the number of cells needed for a given screen, the cell coverage required for large-scale CRISPR screens still poses technical hurdles to screen in more challenging systems, such as iPSC-derived and primary cells. A major factor that influences cell coverage is screening library uniformity, as larger variation in individual guide RNA abundance requires higher cell coverage to reliably measure low-abundance guides. In this work, we have systematically optimized guide RNA cloning procedures to decrease bias. We implement these protocols to demonstrate that CRISPRi screens using 10-fold fewer cells than the current standard provides equivalent statistically significant hit-calling results to screens run at higher coverage, opening the possibility of conducting genome-wide and other large-scale CRISPR screens in technically challenging models.
]]></description>
<dc:creator>Heo, S.-J.</dc:creator>
<dc:creator>Enriquez, L. D.</dc:creator>
<dc:creator>Federman, S.</dc:creator>
<dc:creator>Chang, A. Y.</dc:creator>
<dc:creator>Mace, R.</dc:creator>
<dc:creator>Sevade, K.</dc:creator>
<dc:creator>Nguyen, P.</dc:creator>
<dc:creator>Litterman, A. J.</dc:creator>
<dc:creator>Shafer, S.</dc:creator>
<dc:creator>Przybyla, L.</dc:creator>
<dc:creator>Chow, E. D.</dc:creator>
<dc:date>2022-12-23</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521524</dc:identifier>
<dc:title><![CDATA[Optimized CRISPR guide RNA library cloning reduces skew and enables more compact genetic screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.22.521678v1?rss=1">
<title>
<![CDATA[
Uncovering context-specific genetic-regulation of gene expression from single-cell RNA-sequencing using latent-factor models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.22.521678v1?rss=1</link>
<description><![CDATA[
Genetic regulation of gene expression is a complex process, with genetic effects known to vary across cellular contexts such as cell types and environmental conditions. We developed SURGE, a method for unsupervised discovery of context-specific expression quantitative trait loci (eQTLs) from single-cell transcriptomic data. This allows discovery of the contexts or cell types modulating genetic regulation without prior knowledge. Applied to peripheral blood single-cell eQTL data, SURGE contexts capture continuous representations of distinct cell types and groupings of biologically related cell types. We demonstrate the disease-relevance of SURGE context-specific eQTLs using colocalization analysis and stratified LD-score regression.
]]></description>
<dc:creator>Strober, B. J.</dc:creator>
<dc:creator>Tayeb, K.</dc:creator>
<dc:creator>Popp, J.</dc:creator>
<dc:creator>Qi, G.</dc:creator>
<dc:creator>Gordon, M. G.</dc:creator>
<dc:creator>Perez, R.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Battle, A.</dc:creator>
<dc:date>2022-12-23</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521678</dc:identifier>
<dc:title><![CDATA[Uncovering context-specific genetic-regulation of gene expression from single-cell RNA-sequencing using latent-factor models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.23.521842v1?rss=1">
<title>
<![CDATA[
Epigenetic reprogramming shapes the cellular landscape of schwannoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.23.521842v1?rss=1</link>
<description><![CDATA[
Cell state evolution underlies tumor development and response to therapy1, but mechanisms specifying cancer cell states and intratumor heterogeneity are incompletely understood. Schwannomas are the most common tumors of the peripheral nervous system and are treated with surgery and ionizing radiation2-5. Schwannomas can oscillate in size for many years after radiotherapy6,7, suggesting treatment may reprogram schwannoma cells or the tumor microenvironment. Here we show epigenetic reprogramming shapes the cellular landscape of schwannomas. We find schwannomas are comprised of 2 molecular groups distinguished by reactivation of neural crest development pathways or misactivation of nerve injury mechanisms that specify cancer cell states and the architecture of the tumor immune microenvironment. Schwannoma molecular groups can arise independently, but ionizing radiation is sufficient for epigenetic reprogramming of neural crest to immune-enriched schwannoma by remodeling chromatin accessibility, gene expression, and metabolism to drive schwannoma cell state evolution and immune cell infiltration. To define functional genomic mechanisms underlying epigenetic reprograming of schwannomas, we develop a technique for simultaneous interrogation of chromatin accessibility and gene expression coupled with genetic and therapeutic perturbations in single-nuclei. Our results elucidate a framework for understanding epigenetic drivers of cancer evolution and establish a paradigm of epigenetic reprograming of cancer in response to radiotherapy.
]]></description>
<dc:creator>Liu, S. J.</dc:creator>
<dc:creator>Casey-Clyde, T.</dc:creator>
<dc:creator>Cho, N. W.</dc:creator>
<dc:creator>Swinderman, J.</dc:creator>
<dc:creator>Pekmezci, M.</dc:creator>
<dc:creator>Dougherty, M. C.</dc:creator>
<dc:creator>Foster, K.</dc:creator>
<dc:creator>Chen, W. C.</dc:creator>
<dc:creator>Villanueva-Meyer, J. E.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Vasudevan, H. N.</dc:creator>
<dc:creator>Choudhury, A.</dc:creator>
<dc:creator>Breshears, J. D.</dc:creator>
<dc:creator>Lang, U. E.</dc:creator>
<dc:creator>Eaton, C. D.</dc:creator>
<dc:creator>Hiam-Galvez, K. J.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Chen, K.-H.</dc:creator>
<dc:creator>Lien, B. V.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Braunstein, S. E.</dc:creator>
<dc:creator>Sneed, P. K.</dc:creator>
<dc:creator>Magill, S. T.</dc:creator>
<dc:creator>Lim, D. A.</dc:creator>
<dc:creator>McDermott, M. W.</dc:creator>
<dc:creator>Berger, M. S.</dc:creator>
<dc:creator>Perry, A.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Hansen, M.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:creator>Theodosopoulos, P. V.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:date>2022-12-24</dc:date>
<dc:identifier>doi:10.1101/2022.12.23.521842</dc:identifier>
<dc:title><![CDATA[Epigenetic reprogramming shapes the cellular landscape of schwannoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.27.521999v1?rss=1">
<title>
<![CDATA[
A 3D Clinical Face Phenotype Space of Genetic Syndromes using a Triplet-Based Singular Geometric Autoencoder 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.27.521999v1?rss=1</link>
<description><![CDATA[
Clinical diagnosis of syndromes benefits strongly from objective facial phenotyping. This study introduces a novel approach to enhance clinical diagnosis through the development and exploration of a low-dimensional metric space referred to as the clinical face phenotypic space (CFPS). As a facial matching tool for clinical genetics, such CFPS can enhance clinical diagnosis. It helps to interpret facial dysmorphisms of a subject by placing them within the space of known dysmorphisms. In this paper, a triplet loss-based autoencoder developed by geometric deep learning (GDL) is trained using multi-task learning, which combines supervised and unsupervised learning approaches. Experiments are designed to illustrate the following properties of CFPSs that can aid clinicians in narrowing down their search space: a CFPS can 1) classify syndromes accurately, 2) generalize to novel syndromes, and 3) preserve the relatedness of genetic diseases, meaning that clusters of phenotypically similar disorders reflect functional relationships between genes. The proposed model consists of three main components: an encoder based on GDL optimizing distances between groups of individuals in the CFPS, a decoder enhancing classification by reconstructing faces, and a singular value decomposition layer maintaining orthogonality and optimal variance distribution across dimensions. This allows for the selection of an optimal number of CFPS dimensions as well as improving the classification capacity of the CFPS.
]]></description>
<dc:creator>Mahdi, S. S.</dc:creator>
<dc:creator>Matthews, H.</dc:creator>
<dc:creator>Vanneste, M.</dc:creator>
<dc:creator>Nauwelaers, N.</dc:creator>
<dc:creator>Gong, S.</dc:creator>
<dc:creator>Bouritsas, G.</dc:creator>
<dc:creator>Baynam, G. S.</dc:creator>
<dc:creator>Hammond, P.</dc:creator>
<dc:creator>Spritz, R.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Hallgrimsson, B. D.</dc:creator>
<dc:creator>Peeters, H.</dc:creator>
<dc:creator>Claes, P.</dc:creator>
<dc:date>2022-12-27</dc:date>
<dc:identifier>doi:10.1101/2022.12.27.521999</dc:identifier>
<dc:title><![CDATA[A 3D Clinical Face Phenotype Space of Genetic Syndromes using a Triplet-Based Singular Geometric Autoencoder]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.29.522180v1?rss=1">
<title>
<![CDATA[
Deficiency in Galectin-3, -8, and -9 impairs immunity to chronic Mycobacterium tuberculosis infection but not acute infection with multiple intracellular pathogens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.29.522180v1?rss=1</link>
<description><![CDATA[
Macrophages employ an array of pattern recognition receptors to detect and eliminate intracellular pathogens that access the cytosol. The cytosolic carbohydrate sensors Galectin-3, -8, and -9 (Gal-3, Gal-8, and Gal-9) recognize damaged pathogen-containing phagosomes, and Gal-3 and Gal-8 are reported to restrict bacterial growth via autophagy in cultured cells. However, the contribution of these galectins to host resistance during bacterial infection remains unclear. We found that Gal-9 binds directly to Mycobacterium tuberculosis (Mtb) and Salmonella enterica serovar Typhimurium (Stm) and localizes to Mtb in macrophages. To determine the combined contribution of membrane damage-sensing galectins to immunity in vivo, we generated Gal-3, -8, and - 9 triple knockout (TKO) mice. Mtb infection of primary macrophages from TKO mice resulted in defective lysosomal trafficking but normal bacterial replication. Surprisingly, these mice had no discernable defect in resistance to acute infection with Mtb, Stm or Listeria monocytogenes, and had only modest impairments in bacterial growth restriction and CD4 T cell activation during chronic Mtb infection. Collectively, these findings indicate that while Gal-3, -8, and -9 respond to an array of intracellular pathogens, together these membrane damage-sensing galectins play a limited role in host resistance to bacterial infection.

Author SummaryIntracellular bacterial pathogens cause many of the worlds most deadly infectious diseases. A common requirement for nearly all intracellular pathogens is the ability to damage the endomembrane compartments in which they reside, which allows pathogens access to the nutrient-rich cytosol of the host. However, membrane damage also creates a "pattern of pathogenesis" that triggers antimicrobial immune responses. Galectin-3, -8, and -9 (Gal-3, Gal-8, and Gal-9) act as a surveillance system for membrane damage and Gal-3 and Gal-8 inhibit bacterial growth by activating autophagy, a cellular pathway that can capture cytosolic bacteria and degrade them in lysosomes. Membrane damage-sensing galectins were hypothesized to promote bacterial killing during acute infection yet their role in the immune response of an infected animal remains unclear. Here, we show that mice deficient for Gal-3, -8, and -9 had no defects in resistance to acute infection with the pathogens Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and Mycobacterium tuberculosis (Mtb), and were only modestly susceptible to chronic Mtb infection. Our data suggest that Gal-3, -8 and -9 are not critical for innate immune responses during acute infection and may play a more prominent role in the adaptive immune response. These results broaden our understanding of the role of membrane damage-sensing pathways in host defense against bacterial infection.
]]></description>
<dc:creator>Morrison, H. M.</dc:creator>
<dc:creator>Craft, J.</dc:creator>
<dc:creator>Rivera-Lugo, R.</dc:creator>
<dc:creator>Johnson, J. R.</dc:creator>
<dc:creator>Golovkine, G. R.</dc:creator>
<dc:creator>Dodd, C. E.</dc:creator>
<dc:creator>Van Dis, E.</dc:creator>
<dc:creator>Beatty, W. L.</dc:creator>
<dc:creator>Margolis, S. R.</dc:creator>
<dc:creator>Repasy, T.</dc:creator>
<dc:creator>Shaker, I.</dc:creator>
<dc:creator>Lee, A. Y.</dc:creator>
<dc:creator>Vance, R. E.</dc:creator>
<dc:creator>Stanley, S. A.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Portnoy, D. A.</dc:creator>
<dc:creator>Penn, B. H.</dc:creator>
<dc:creator>Cox, J. S.</dc:creator>
<dc:date>2022-12-29</dc:date>
<dc:identifier>doi:10.1101/2022.12.29.522180</dc:identifier>
<dc:title><![CDATA[Deficiency in Galectin-3, -8, and -9 impairs immunity to chronic Mycobacterium tuberculosis infection but not acute infection with multiple intracellular pathogens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.30.521706v1?rss=1">
<title>
<![CDATA[
Rac negative feedback links local PIP3 rate-of-change to dynamic control of neutrophil guidance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.30.521706v1?rss=1</link>
<description><![CDATA[
To migrate efficiently, neutrophils must polarize their cytoskeletal regulators along a single axis of motion. This polarization process is thought to be mediated through local positive feedback that amplifies leading edge signals and global negative feedback that enables sites of positive feedback to compete for dominance. Though this two-component model efficiently establishes cell polarity, it has potential limitations, including a tendency to "lock" onto a particular direction, limiting the ability of cells to reorient. We use spatially-defined optogenetic control of a leading edge organizer (PI3K) to probe how cells balance "decisiveness" needed to polarize in a single direction with the flexibility needed to respond to new cues. Underlying this balancing act is a local Rac inhibitor that destabilizes the leading edge to promote exploration. We show that this local inhibitor enables cells to process input signal dynamics, linking front stability and orientation to local temporal increases in input signals.
]]></description>
<dc:creator>Town, J. P.</dc:creator>
<dc:creator>Weiner, O. P.</dc:creator>
<dc:date>2022-12-31</dc:date>
<dc:identifier>doi:10.1101/2022.12.30.521706</dc:identifier>
<dc:title><![CDATA[Rac negative feedback links local PIP3 rate-of-change to dynamic control of neutrophil guidance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.05.522832v1?rss=1">
<title>
<![CDATA[
Highly multiplexed ddPCR-amplicon sequencing reveals strong Plasmodium falciparum population structure and isolated populations amenable to local elimination efforts in Zanzibar 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.05.522832v1?rss=1</link>
<description><![CDATA[
Over the past 15 years, Zanzibar has made great strides towards malaria elimination; yet progress has stalled. Parasite genetic data of Plasmodium falciparum may inform strategies for malaria elimination by helping to identify contributory factors to parasite persistence. Here we elucidate fine-scale parasite population structure and infer relatedness and connectivity of infections using an identity-by-descent (IBD) approach. We sequenced 518 P. falciparum samples from 5 districts covering both main islands using a novel, highly multiplexed droplet digital PCR (ddPCR)-based amplicon deep sequencing method targeting 35 microhaplotypes and drug-resistance loci. Despite high genetic diversity, we observe strong fine-scale spatial and temporal structure of local parasite populations, including isolated populations on Pemba Island and genetically admixed populations on Unguja Island, providing evidence of ongoing local transmission. We observe a high proportion of highly related parasites in individuals living closer together, including between clinical index cases and the mostly asymptomatic cases surrounding them, consistent with isolation-by-distance. We identify a substantial fraction (2.9%) of related parasite pairs between Zanzibar, and mainland Tanzania and Kenya, consistent with recent importation. We identify haplotypes known to confer resistance to known antimalarials in all districts, including multidrug-resistant parasites, but most parasites remain sensitive to current first-line treatments. Our study provides a high-resolution view of parasite genetic structure across the Zanzibar archipelago and reveals actionable patterns, including isolated parasite populations, which may be prioritized for malaria elimination.
]]></description>
<dc:creator>Holzschuh, A.</dc:creator>
<dc:creator>Lerch, A.</dc:creator>
<dc:creator>Gerlovina, I.</dc:creator>
<dc:creator>Fakih, B. S.</dc:creator>
<dc:creator>Al-Mafazy, A.-W.</dc:creator>
<dc:creator>Reaves, E. J.</dc:creator>
<dc:creator>Ali, A.</dc:creator>
<dc:creator>Abbas, F.</dc:creator>
<dc:creator>Ali, M. H.</dc:creator>
<dc:creator>Ali, M. A.</dc:creator>
<dc:creator>Hetzel, M. W.</dc:creator>
<dc:creator>Yukich, J. O.</dc:creator>
<dc:creator>Koepfli, C.</dc:creator>
<dc:date>2023-01-06</dc:date>
<dc:identifier>doi:10.1101/2023.01.05.522832</dc:identifier>
<dc:title><![CDATA[Highly multiplexed ddPCR-amplicon sequencing reveals strong Plasmodium falciparum population structure and isolated populations amenable to local elimination efforts in Zanzibar]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.05.522921v1?rss=1">
<title>
<![CDATA[
Phenotypic subtyping via contrastive learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.05.522921v1?rss=1</link>
<description><![CDATA[
Defining and accounting for subphenotypic structure has the potential to increase statistical power and provide a deeper understanding of the heterogeneity in the molecular basis of complex disease. Existing phenotype subtyping methods primarily rely on clinically observed heterogeneity or metadata clustering. However, they generally tend to capture the dominant sources of variation in the data, which often originate from variation that is not descriptive of the mechanistic heterogeneity of the phenotype of interest; in fact, such dominant sources of variation, such as population structure or technical variation, are, in general, expected to be independent of subphenotypic structure. We instead aim to find a subspace with signal that is unique to a group of samples for which we believe that subphenotypic variation exists (e.g., cases of a disease). To that end, we introduce Phenotype Aware Components Analysis (PACA), a contrastive learning approach leveraging canonical correlation analysis to robustly capture weak sources of subphenotypic variation. In the context of disease, PACA learns a gradient of variation unique to cases in a given dataset, while leveraging control samples for accounting for variation and imbalances of biological and technical confounders between cases and controls. We evaluated PACA using an extensive simulation study, as well as on various subtyping tasks using genotypes, transcriptomics, and DNA methylation data. Our results provide multiple strong evidence that PACA allows us to robustly capture weak unknown variation of interest while being calibrated and well-powered, far superseding the performance of alternative methods. This renders PACA as a state-of-the-art tool for defining de novo subtypes that are more likely to reflect molecular heterogeneity, especially in challenging cases where the phenotypic heterogeneity may be masked by a myriad of strong unrelated effects in the data.

Code AvailabilityPACA is available as an open source R package on GitHub: https://github.com/Adigorla/PACA
]]></description>
<dc:creator>Gorla, A.</dc:creator>
<dc:creator>Sankararaman, S.</dc:creator>
<dc:creator>Burchard, E.</dc:creator>
<dc:creator>Flint, J.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:creator>Rahmani, E.</dc:creator>
<dc:date>2023-01-06</dc:date>
<dc:identifier>doi:10.1101/2023.01.05.522921</dc:identifier>
<dc:title><![CDATA[Phenotypic subtyping via contrastive learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.05.522960v1?rss=1">
<title>
<![CDATA[
Graph Convolutional learning of Multimodal Brain Connectome Data for Schizophrenia Classification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.05.522960v1?rss=1</link>
<description><![CDATA[
The long term goal of this work is to develop powerful tools for brain network analysis in order to study structural and functional connectivity abnormalities in psychiatric disorders like schizophrenia. Graph convolutional neural networks (GCNN) are quite effective for learning complex discriminate features in graph-structured data. Here, we explore the GCNN to learn the discriminating features in multimodal human brain connectomes for the purpose of schizophrenia disorder classification. In particular, we train and validate a network using both structural connectivity graphs obtained from diffusion tensor imaging data and functional connectivity from functional magnetic resonance imaging data.We compare the GCNN method with a support vector machine based classifier and other popular classification benchmarks. We demonstrate that the proposed graph convolution method has the best performance compared to existing benchmarks with F1 scores of 0.75 for schizophrenia classification. This demonstrates the potential of this approach for multimodal diagnosis and prognosis in mental health disorders.
]]></description>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Bhargava, E.</dc:creator>
<dc:creator>Lin, C.-T.</dc:creator>
<dc:creator>Nagarajan, S. S.</dc:creator>
<dc:date>2023-01-06</dc:date>
<dc:identifier>doi:10.1101/2023.01.05.522960</dc:identifier>
<dc:title><![CDATA[Graph Convolutional learning of Multimodal Brain Connectome Data for Schizophrenia Classification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.06.522937v1?rss=1">
<title>
<![CDATA[
Chromatin Remodeling Drives Immune-Fibroblast Crosstalk in Heart Failure Pathogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.06.522937v1?rss=1</link>
<description><![CDATA[
Chronic inflammation and tissue fibrosis are common stress responses that worsen organ function, yet the molecular mechanisms governing their crosstalk are poorly understood. In diseased organs, stress-induced changes in gene expression fuel maladaptive cell state transitions and pathological interaction between diverse cellular compartments. Although chronic fibroblast activation worsens dysfunction of lung, liver, kidney, and heart, and exacerbates many cancers, the stress-sensing mechanisms initiating the transcriptional activation of fibroblasts are not well understood. Here, we show that conditional deletion of the transcription co-activator Brd4 in Cx3cr1-positive myeloid cells ameliorates heart failure and is associated with a dramatic reduction in fibroblast activation. Analysis of single-cell chromatin accessibility and BRD4 occupancy in vivo in Cx3cr1-positive cells identified a large enhancer proximal to Interleukin-1 beta (Il1b), and a series of CRISPR deletions revealed the precise stress-dependent regulatory element that controlled expression of Il1b in disease. Secreted IL1B functioned non-cell autonomously to activate a p65/RELA-dependent enhancer near the transcription factor MEOX1, resulting in a profibrotic response in human cardiac fibroblasts. In vivo, antibody-mediated IL1B neutralization prevented stress-induced expression of MEOX1, inhibited fibroblast activation, and improved cardiac function in heart failure. The elucidation of BRD4-dependent crosstalk between a specific immune cell subset and fibroblasts through IL1B provides new therapeutic strategies for heart disease and other disorders of chronic inflammation and maladaptive tissue remodeling.
]]></description>
<dc:creator>Alexanian, M.</dc:creator>
<dc:creator>Padmanabhan, A.</dc:creator>
<dc:creator>Nishino, T.</dc:creator>
<dc:creator>Travers, J. G.</dc:creator>
<dc:creator>Ye, L.</dc:creator>
<dc:creator>Lee, C. Y.</dc:creator>
<dc:creator>Sadagopan, N.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Pelonero, A.</dc:creator>
<dc:creator>Auclair, K.</dc:creator>
<dc:creator>Zhu, A.</dc:creator>
<dc:creator>Gonzalez Teran, B.</dc:creator>
<dc:creator>Flanigan, W.</dc:creator>
<dc:creator>Kim, C. K.-S.</dc:creator>
<dc:creator>Lumbao-Conradson, K.</dc:creator>
<dc:creator>Costa, M.</dc:creator>
<dc:creator>Jain, R.</dc:creator>
<dc:creator>Charo, I.</dc:creator>
<dc:creator>Haldar, S. M.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Vagnozzi, R. J.</dc:creator>
<dc:creator>McKinsey, T. A.</dc:creator>
<dc:creator>Przytycki, P. F.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:date>2023-01-07</dc:date>
<dc:identifier>doi:10.1101/2023.01.06.522937</dc:identifier>
<dc:title><![CDATA[Chromatin Remodeling Drives Immune-Fibroblast Crosstalk in Heart Failure Pathogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.06.523027v1?rss=1">
<title>
<![CDATA[
M. tuberculosis antigen-responsive IL17+ CD4 T cells are disproportionately spared in ART-suppressed HIV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.06.523027v1?rss=1</link>
<description><![CDATA[
BackgroundInterleukin 17 producing CD4 T cells contribute to the control of Mycobacterium tuberculosis (Mtb) infection in humans; whether infection with Human Immunodeficiency Virus (HIV) disproportionately affects distinct Th17 cell subsets that respond to Mtb is incompletely defined.

MethodsWe performed high-definition characterization of circulating Mtb-specific Th17 cells by spectral flow cytometry in people with latent TB and treated HIV (HIV-ART). We also measured kynurenine pathway activity by LC/MS on plasma and tested the hypothesis that tryptophan catabolism influences Th17 cell frequencies in this context.

ResultsWe identified two subsets of Th17 cells: subset 1 defined as CD4+V7.2-CD161+CD26+ and subset 2 defined as CD4+V7.2-CCR6+CXCR3- cells of which subset 1 was significantly reduced in LTBI with HIV-ART, yet Mtb-responsive IL17-producing CD4 T cells were preserved; we found that IL17-producing CD4 T cells dominate the response to Mtb antigen but not CMV antigen or staphylococcal enterotoxin B (SEB); and tryptophan catabolism negatively correlates with both subset 1 and subset 2 Th17 cell frequencies.

ConclusionsWe found differential effects of ART-suppressed HIV on distinct subsets of Th17 cells, that IL17-producing CD4 T cells dominate responses to Mtb but not CMV antigen or SEB, and that kynurenine pathway activity is associated with decreases of circulating Th17 cells that may contribute to tuberculosis immunity.
]]></description>
<dc:creator>Ogongo, P.</dc:creator>
<dc:creator>Tran, A.</dc:creator>
<dc:creator>Marzan, F.</dc:creator>
<dc:creator>Gingrich, D.</dc:creator>
<dc:creator>Krone, M.</dc:creator>
<dc:creator>Aweeka, F.</dc:creator>
<dc:creator>Lindestam Arlehamn, C. S.</dc:creator>
<dc:creator>Martin, J. N.</dc:creator>
<dc:creator>Deeks, S.</dc:creator>
<dc:creator>Hunt, P. W.</dc:creator>
<dc:creator>Ernst, J. D.</dc:creator>
<dc:date>2023-01-07</dc:date>
<dc:identifier>doi:10.1101/2023.01.06.523027</dc:identifier>
<dc:title><![CDATA[M. tuberculosis antigen-responsive IL17+ CD4 T cells are disproportionately spared in ART-suppressed HIV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.07.522948v1?rss=1">
<title>
<![CDATA[
INKILN is a novel long noncoding RNA promoting vascular smooth muscle inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.07.522948v1?rss=1</link>
<description><![CDATA[
BackgroundActivation of vascular smooth muscle cells (VSMCs) inflammation is vital to initiate vascular disease. However, the role of human-specific long noncoding RNAs (lncRNAs) in VSMC inflammation is poorly understood.

MethodsBulk RNA-seq in differentiated human VSMCs revealed a novel human-specific lncRNA called INflammatory MKL1 Interacting Long Noncoding RNA (INKILN). INKILN expression was assessed in multiple in vitro and ex vivo models of VSMC phenotypic modulation and human atherosclerosis and abdominal aortic aneurysm (AAA) samples. The transcriptional regulation of INKILN was determined through luciferase reporter system and chromatin immunoprecipitation assay. Both loss- and gain-of-function approaches and multiple RNA-protein and protein-protein interaction assays were utilized to uncover the role of INKILN in VSMC proinflammatory gene program and underlying mechanisms. Bacterial Artificial Chromosome (BAC) transgenic (Tg) mice were utilized to study INKLIN expression and function in ligation injury-induced neointimal formation.

ResultsINKILN expression is downregulated in contractile VSMCs and induced by human atherosclerosis and abdominal aortic aneurysm. INKILN is transcriptionally activated by the p65 pathway, partially through a predicted NF-{kappa}B site within its proximal promoter. INKILN activates the proinflammatory gene expression in cultured human VSMCs and ex vivo cultured vessels. Mechanistically, INKILN physically interacts with and stabilizes MKL1, a key activator of VSMC inflammation through the p65/NF-{kappa}B pathway. INKILN depletion blocks ILI{beta}-induced nuclear localization of both p65 and MKL1. Knockdown of INKILN abolishes the physical interaction between p65 and MKL1, and the luciferase activity of an NF-{kappa}B reporter. Further, INKILN knockdown enhances MKL1 ubiquitination, likely through the reduced physical interaction with the deubiquitinating enzyme, USP10. INKILN is induced in injured carotid arteries and exacerbates ligation injury-induced neointimal formation in BAC Tg mice.

ConclusionsThese findings elucidate an important pathway of VSMC inflammation involving an INKILN/MKL1/USP10 regulatory axis. Human BAC Tg mice offer a novel and physiologically relevant approach for investigating human-specific lncRNAs under vascular disease conditions.
]]></description>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Deng, L.</dc:creator>
<dc:creator>Ishimwe, N.</dc:creator>
<dc:creator>Pauli, J.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Shan, S.</dc:creator>
<dc:creator>Kempf, W.</dc:creator>
<dc:creator>Ballantyne, M. D.</dc:creator>
<dc:creator>Kim, D.</dc:creator>
<dc:creator>Lyu, Q.</dc:creator>
<dc:creator>Bennett, M.</dc:creator>
<dc:creator>Rodor, J.</dc:creator>
<dc:creator>Turner, A. W.</dc:creator>
<dc:creator>Lu, Y. W.</dc:creator>
<dc:creator>Gao, P.</dc:creator>
<dc:creator>Choi, M.</dc:creator>
<dc:creator>Warthi, G.</dc:creator>
<dc:creator>Kim, H. W.</dc:creator>
<dc:creator>Barroso, M. M.</dc:creator>
<dc:creator>Bryant, W. B.</dc:creator>
<dc:creator>Miller, C. L.</dc:creator>
<dc:creator>Weintraub, N. L.</dc:creator>
<dc:creator>Maegdefessel, L.</dc:creator>
<dc:creator>Miano, J. M.</dc:creator>
<dc:creator>Baker, A. H.</dc:creator>
<dc:creator>Long, X.</dc:creator>
<dc:date>2023-01-08</dc:date>
<dc:identifier>doi:10.1101/2023.01.07.522948</dc:identifier>
<dc:title><![CDATA[INKILN is a novel long noncoding RNA promoting vascular smooth muscle inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.09.523344v1?rss=1">
<title>
<![CDATA[
Oncogene-like addiction to aneuploidy in human cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.09.523344v1?rss=1</link>
<description><![CDATA[
Most cancers exhibit aneuploidy, but its functional significance in tumor development is controversial. Here, we describe ReDACT (Restoring Disomy in Aneuploid cells using CRISPR Targeting), a set of chromosome engineering tools that allow us to eliminate specific aneuploidies from cancer genomes. Using ReDACT, we created a panel of isogenic cells that have or lack common aneuploidies, and we demonstrate that trisomy of chromosome 1q is required for malignant growth in cancers harboring this alteration. Mechanistically, gaining chromosome 1q increases the expression of MDM4 and suppresses TP53 signaling, and we show that TP53 mutations are mutually-exclusive with 1q aneuploidy in human cancers. Thus, specific aneuploidies play essential roles in tumorigenesis, raising the possibility that targeting these "aneuploidy addictions" could represent a novel approach for cancer treatment.
]]></description>
<dc:creator>Girish, V.</dc:creator>
<dc:creator>Lakhani, A. A.</dc:creator>
<dc:creator>Scaduto, C. M.</dc:creator>
<dc:creator>Thompson, S. L.</dc:creator>
<dc:creator>Brown, L. M.</dc:creator>
<dc:creator>Hagenson, R. A.</dc:creator>
<dc:creator>Sausville, E. L.</dc:creator>
<dc:creator>Mendelson, B. E.</dc:creator>
<dc:creator>Lukow, D. A.</dc:creator>
<dc:creator>Yuan, M. L.</dc:creator>
<dc:creator>Kandikuppa, P. K.</dc:creator>
<dc:creator>Stevens, E. C.</dc:creator>
<dc:creator>Lee, S. N.</dc:creator>
<dc:creator>Salovska, B.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Smith, J. C.</dc:creator>
<dc:creator>Taylor, A. M.</dc:creator>
<dc:creator>Martienssen, R. A.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Sun, R.</dc:creator>
<dc:creator>Sheltzer, J. M.</dc:creator>
<dc:date>2023-01-10</dc:date>
<dc:identifier>doi:10.1101/2023.01.09.523344</dc:identifier>
<dc:title><![CDATA[Oncogene-like addiction to aneuploidy in human cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.09.523364v1?rss=1">
<title>
<![CDATA[
Genome wide-analysis of anterior-posterior mRNA localization in Stentor coeruleus reveals a role for the microtubule cytoskeleton 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.09.523364v1?rss=1</link>
<description><![CDATA[
Cells have complex and beautiful structures that are important for their function. However, understanding the molecular mechanisms that produce these structures is a challenging problem due to the gap in size scales between molecular interactions and cellular structures. The giant ciliate Stentor coeruleus is a unicellular model organism whose large size, reproducible structure, and ability to heal wounds and regenerate have historically allowed the formation of structure in a single cell to be addressed using methods of experimental embryology. Such studies have shown that specific cellular structures, such as the oral apparatus, always form in particular regions of the cell, which raises the question: what is the source of positional information within this organism? By analogy with embryonic development, in which regionalized mRNA is often used to mark position, we asked whether specific regionalized mRNAs might mark position along the anterior-posterior axis of Stentor. By physically bisecting cells and conducting bulk RNA sequencing, we were able to identify sets of messages enriched in either the anterior or posterior half and show that RNAi-mediated knockdown of posterior-enriched transcripts corresponding to MYB genes results in a cells inability to regenerate the posterior portion of the cell body. We then conducted half-cell RNA-sequencing in paired anteriors and posteriors of cells in which the microtubule cytoskeleton was disrupted by RNAi of {beta}-tubulin or dynein intermediate chains. We found that many messages either lost their regionalized distribution or switched to an opposite distribution, such that anterior-enriched messages in control became posterior-enriched in the RNAi cells, or vice versa. This study indicates that mRNA can be regionalized within a single giant cell and that microtubules may play a role, possibly by serving as tracks for the movement of the messages.
]]></description>
<dc:creator>Albright, A. R.</dc:creator>
<dc:creator>Angeles-Albores, D.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2023-01-10</dc:date>
<dc:identifier>doi:10.1101/2023.01.09.523364</dc:identifier>
<dc:title><![CDATA[Genome wide-analysis of anterior-posterior mRNA localization in Stentor coeruleus reveals a role for the microtubule cytoskeleton]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.10.523504v1?rss=1">
<title>
<![CDATA[
Cryo-EM reveals how Hsp90 and FKBP immunophilins co-regulate the Glucocorticoid Receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.10.523504v1?rss=1</link>
<description><![CDATA[
Hsp90 is an essential molecular chaperone responsible for the folding and activation of hundreds of  client proteins, including the glucocorticoid receptor (GR)1-3. Previously, we revealed that GR ligand binding activity is inhibited by Hsp70 and restored by Hsp90, aided by co-chaperones4. We then presented cryo-EM structures mechanistically detailing how Hsp70 and Hsp90 remodel the conformation of GR to regulate ligand binding5,6. In vivo, GR-chaperone complexes are found associated with numerous Hsp90 co-chaperones, but the most enigmatic have been the immunophilins FKBP51 and FKBP52, which further regulate the activity of GR and other steroid receptors7-9. A molecular understanding of how FKBP51 and FKBP52 integrate with the GR chaperone cycle to differentially regulate GR activation in vivo is lacking due to difficulties reconstituting these interactions. Here, we present a 3.01 [A] cryo-EM structure of the GR:Hsp90:FKBP52 complex, revealing, for the first time, that FKBP52 directly binds to the folded, ligand-bound GR using three novel interfaces, each of which we demonstrate are critical for FKBP52-dependent potentiation of GR activity in vivo. In addition, we present a 3.23 [A] cryo-EM structure of the GR:Hsp90:FKBP51 complex, which, surprisingly, largely mimics the GR:Hsp90:FKBP52 structure. In both structures, FKBP51 and FKBP52 directly engage the folded GR and unexpectedly facilitate release of p23 through an allosteric mechanism. We also reveal that FKBP52, but not FKBP51, potentiates GR ligand binding in vitro, in a manner dependent on FKBP52-specific interactions. Altogether, we reveal how FKBP51 and FKBP52 integrate into the GR chaperone cycle to advance GR to the next stage of maturation and how FKBP51 and FKBP52 compete for GR:Hsp90 binding, leading to functional antagonism.
]]></description>
<dc:creator>Noddings, C. M.</dc:creator>
<dc:creator>Johnson, J. L.</dc:creator>
<dc:creator>Agard, D. A.</dc:creator>
<dc:date>2023-01-11</dc:date>
<dc:identifier>doi:10.1101/2023.01.10.523504</dc:identifier>
<dc:title><![CDATA[Cryo-EM reveals how Hsp90 and FKBP immunophilins co-regulate the Glucocorticoid Receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.10.523535v1?rss=1">
<title>
<![CDATA[
Differences in expression of tumor suppressor, innate immune, inflammasome, and potassium/gap junction channel host genes significantly predict viral reservoir size during treated HIV infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.10.523535v1?rss=1</link>
<description><![CDATA[
The major barrier to an HIV cure is the persistence of infected cells that evade host immune surveillance despite effective antiretroviral therapy (ART). Most prior host genetic HIV studies have focused on identifying DNA polymorphisms (e.g., CCR5{Delta}32, MHC class I alleles) associated with viral load among untreated "elite controllers" (~1% of HIV+ individuals who are able to control virus without ART). However, there have been few studies evaluating host genetic predictors of viral control for the majority of people living with HIV (PLWH) on ART. We performed host RNA sequencing and HIV reservoir quantification (total DNA, unspliced RNA, intact DNA) from peripheral CD4+ T cells from 191 HIV+ ART-suppressed non-controllers. Multivariate models included covariates for timing of ART initiation, nadir CD4+ count, age, sex, and ancestry. Lower HIV total DNA (an estimate of the total reservoir) was associated with upregulation of tumor suppressor genes NBL1 (q=0.012) and P3H3 (q=0.012). Higher HIV unspliced RNA (an estimate of residual HIV transcription) was associated with downregulation of several host genes involving inflammasome (IL1A, CSF3, TNFAIP5, TNFAIP6, TNFAIP9, CXCL3, CXCL10) and innate immune (TLR7) signaling, as well as novel associations with potassium (KCNJ2) and gap junction (GJB2) channels, all q<0.05. Gene set enrichment analyses identified significant associations with TLR4/microbial translocation (q=0.006), IL-1{beta}/NRLP3 inflammasome (q=0.008), and IL-10 (q=0.037) signaling. HIV intact DNA (an estimate of the "replication-competent" reservoir) demonstrated trends with thrombin degradation (PLGLB1) and glucose metabolism (AGL) genes, but data were (HIV intact DNA detected in only 42% of participants). Our findings demonstrate that among treated PLWH, that inflammation, innate immune responses, bacterial translocation, and tumor suppression/cell proliferation host signaling play a key role in the maintenance of the HIV reservoir during ART. Further data are needed to validate these findings, including functional genomic studies, and expanded epidemiologic studies in female, non-European cohorts.

Author SummaryAlthough lifelong HIV antiretroviral therapy (ART) suppresses virus, the major barrier to an HIV cure is the persistence of infected cells that evade host immune surveillance despite effective ART, "the HIV reservoir." HIV eradication strategies have focused on eliminating residual virus to allow for HIV remission, but HIV cure trials to date have thus far failed to show a clinically meaningful reduction in the HIV reservoir. There is an urgent need for a better understanding of the host-viral dynamics during ART suppression to identify potential novel therapeutic targets for HIV cure. This is the first epidemiologic host gene expression study to demonstrate a significant link between HIV reservoir size and several well-known immunologic pathways (e.g., IL-1{beta}, TLR7, TNF- signaling pathways), as well as novel associations with potassium and gap junction channels (Kir2.1, connexin 26). Further data are needed to validate these findings, including functional genomic studies and expanded epidemiologic studies in female, non-European cohorts.
]]></description>
<dc:creator>Dwivedi, A. K.</dc:creator>
<dc:creator>Siegel, D. A.</dc:creator>
<dc:creator>Thanh, C.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Hobbs, K. S.</dc:creator>
<dc:creator>Pan, T.</dc:creator>
<dc:creator>Gibson, E. A.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Hecht, F.</dc:creator>
<dc:creator>Pilcher, C.</dc:creator>
<dc:creator>Milush, J.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Huang, M.-L.</dc:creator>
<dc:creator>Levy, C. N.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Hladik, F.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Henrich, T. J.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Lee, S. A.</dc:creator>
<dc:date>2023-01-11</dc:date>
<dc:identifier>doi:10.1101/2023.01.10.523535</dc:identifier>
<dc:title><![CDATA[Differences in expression of tumor suppressor, innate immune, inflammasome, and potassium/gap junction channel host genes significantly predict viral reservoir size during treated HIV infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.10.523506v1?rss=1">
<title>
<![CDATA[
Cooperative hydrodynamics accompany multicellular-like colonial organization in the unicellular ciliate Stentor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.10.523506v1?rss=1</link>
<description><![CDATA[
Evolution of multicellularity from early unicellular ancestors is arguably one of the most important transitions since the origin of life1,2. Multicellularity is often associated with higher nutrient uptake3, better defense against predation, cell specialization and better division of labor4. While many single-celled organisms exhibit both solitary and colonial existence3,5,6, the organizing principles governing the transition and the benefits endowed are less clear. Using the suspension-feeding unicellular protist Stentor coeruleus, we show that hydrodynamic coupling between proximal neighbors results in faster feeding flows that depend on the separation between individuals. Moreover, we find that the accrued benefits in feeding current enhancement are typically asymmetric- individuals with slower solitary currents gain more from partnering than those with faster currents. We find that colony-formation is ephemeral in Stentor and individuals in colonies are highly dynamic unlike other colony-forming organisms like Volvox carteri 3. Our results demonstrate benefits endowed by the colonial organization in a simple unicellular organism and can potentially provide fundamental insights into the selective forces favoring early evolution of multicellular organization.
]]></description>
<dc:creator>Shekhar, S.</dc:creator>
<dc:creator>Guo, H.</dc:creator>
<dc:creator>Colin, S. P.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:creator>Kanso, E. A.</dc:creator>
<dc:creator>Costello, J. H.</dc:creator>
<dc:date>2023-01-12</dc:date>
<dc:identifier>doi:10.1101/2023.01.10.523506</dc:identifier>
<dc:title><![CDATA[Cooperative hydrodynamics accompany multicellular-like colonial organization in the unicellular ciliate Stentor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.11.523662v1?rss=1">
<title>
<![CDATA[
Selection of optimal cell lines for high-content phenotypic screening 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.11.523662v1?rss=1</link>
<description><![CDATA[
High-content microscopy offers a scalable approach to screen against multiple targets in a single pass. Prior work has focused on methods to select "optimal" cellular readouts in microscopy screens. However, methods to select optimal cell line models have garnered much less attention. Here, we provide a roadmap for how to select the cell line or lines that are best suited to identify bioactive compounds and their mechanism of action (MOA). We test our approach on compounds targeting cancer-relevant pathways, ranking cell lines in two tasks: detecting compound activity ("phenoactivity") and grouping compounds with similar MOA by similar phenotype ("phenosimilarity"). Evaluating six cell lines across 3214 well-annotated compounds, we show that optimal cell line selection depends on both the task of interest (e.g. detecting phenoactivity vs. inferring phenosimilarity) and distribution of MOAs within the compound library. Given a task of interest and set of compounds, we provide a systematic framework for choosing optimal cell line(s). Our framework can be used to reduce the number of cell lines required to identify hits within a compound library and help accelerate the pace of early drug discovery.
]]></description>
<dc:creator>Heinrich, L.</dc:creator>
<dc:creator>Kumbier, K.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Altschuler, S.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:date>2023-01-12</dc:date>
<dc:identifier>doi:10.1101/2023.01.11.523662</dc:identifier>
<dc:title><![CDATA[Selection of optimal cell lines for high-content phenotypic screening]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.12.523677v1?rss=1">
<title>
<![CDATA[
Structure-based discovery of inhibitors of the SARS-CoV-2 Nsp14 N7-methyltransferase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.12.523677v1?rss=1</link>
<description><![CDATA[
An under-explored target for SARS-CoV-2 is non-structural protein 14 (Nsp14), a crucial enzyme for viral replication that catalyzes the methylation of N7-guanosine of the viral RNA at 5'-end; this enables the virus to evade the host immune response by mimicking the eukaryotic post-transcriptional modification mechanism. We sought new inhibitors of the S-adenosyl methionine (SAM)-dependent methyltransferase (MTase) activity of Nsp14 with three large library docking strategies. First, up to 1.1 billion make-on-demand ("tangible") lead-like molecules were docked against the enzymes SAM site, seeking reversible inhibitors. On de novo synthesis and testing, three inhibitors emerged with IC50 values ranging from 6 to 43 M, each with novel chemotypes. Structure-guided optimization and in vitro characterization supported their non-covalent mechanism. In a second strategy, docking a library of 16 million tangible fragments revealed nine new inhibitors with IC50 values ranging from 12 to 341 M and ligand efficiencies from 0.29 to 0.42. In a third strategy, a newly created library of 25 million tangible, virtual electrophiles were docked to covalently modify Cys387 in the SAM binding site. Seven inhibitors emerged with IC50 values ranging from 3.2 to 39 M, the most potent being a reversible aldehyde. Initial optimization of a second series yielded a 7 M acrylamide inhibitor. Three inhibitors characteristic of the new series were tested for selectivity against 30 human protein and RNA MTases, with one showing partial selectivity and one showing high selectivity. Overall, 32 inhibitors encompassing eleven chemotypes had IC50 values <50 M and 5 inhibitors in four chemotypes had IC50 values <10 M. These molecules are among the first non-SAM-like inhibitors of Nsp14, providing multiple starting points for optimizing towards antiviral activity.
]]></description>
<dc:creator>Singh, I.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Fink, E.</dc:creator>
<dc:creator>Chau, I.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Rodriguez-Hernandez, A.</dc:creator>
<dc:creator>Glenn, I.</dc:creator>
<dc:creator>Zapatero-Belinchon, F. J.</dc:creator>
<dc:creator>Rodriguez, M.</dc:creator>
<dc:creator>Devkota, K.</dc:creator>
<dc:creator>Deng, Z.</dc:creator>
<dc:creator>White, K.</dc:creator>
<dc:creator>Wan, X.</dc:creator>
<dc:creator>Tolmachova, N. A.</dc:creator>
<dc:creator>Moroz, Y. S.</dc:creator>
<dc:creator>Kaniskan, H. U.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Gastia-Sastre, A.</dc:creator>
<dc:creator>Jin, J.</dc:creator>
<dc:creator>Fujimori, D. G.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Vedadi, M.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2023-01-13</dc:date>
<dc:identifier>doi:10.1101/2023.01.12.523677</dc:identifier>
<dc:title><![CDATA[Structure-based discovery of inhibitors of the SARS-CoV-2 Nsp14 N7-methyltransferase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.13.523943v1?rss=1">
<title>
<![CDATA[
Genome-wide screens identify SEL1L as an intracellular rheostat controlling collagen turnover 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.13.523943v1?rss=1</link>
<description><![CDATA[
Accumulating evidence has implicated impaired extracellular matrix (ECM) clearance as a key factor in fibrotic disease. Despite decades of research elucidating the effectors of ECM clearance, relatively little is understood regarding the upstream regulation of this process. Collagen is the most abundant constituent of normal and fibrotic ECM in mammalian tissues. Its catabolism occurs through extracellular proteolysis and cell-mediated uptake of collagen fragments for intracellular degradation. Given the paucity of information regarding the regulation of this latter process, we executed unbiased genome-wide screens to understand the molecular underpinnings of cell-mediated collagen clearance. Using this approach, we discovered a previously unappreciated mechanism through which collagen biosynthesis is sensed by cells internally and directly regulates clearance of extracellular collagen. The sensing mechanism is dependent on endoplasmic reticulum-resident protein SEL1L and occurs via a noncanonical function of SEL1L. This pathway functions as a homeostatic negative feedback loop that limits collagen accumulation in tissues. In human fibrotic lung disease, the induction of this collagen clearance pathway by collagen synthesis is impaired, thereby contributing to the pathological accumulation of collagen in lung tissue. Thus cell-autonomous, rheostatic collagen clearance is a previously unidentified pathway of tissue homeostasis.
]]></description>
<dc:creator>Podolsky, M. J.</dc:creator>
<dc:creator>Kheyfets, B.</dc:creator>
<dc:creator>Beigh, A. H.</dc:creator>
<dc:creator>Yang, C. D.</dc:creator>
<dc:creator>Lizama Valenzuela, C.</dc:creator>
<dc:creator>Datta, R.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:creator>McManus, M.</dc:creator>
<dc:creator>Qi, L.</dc:creator>
<dc:creator>Atabai, K.</dc:creator>
<dc:date>2023-01-14</dc:date>
<dc:identifier>doi:10.1101/2023.01.13.523943</dc:identifier>
<dc:title><![CDATA[Genome-wide screens identify SEL1L as an intracellular rheostat controlling collagen turnover]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.13.523996v1?rss=1">
<title>
<![CDATA[
Assembly of gut-derived bacterial communities follows "early-bird" resource utilization dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.13.523996v1?rss=1</link>
<description><![CDATA[
Diet can impact host health through changes to the gut microbiota, yet we lack mechanistic understanding linking nutrient availability and microbiota composition. Here, we use thousands of microbial communities cultured in vitro from human feces to uncover simple assembly rules and develop a predictive model of community composition upon addition of single nutrients from central carbon metabolism to a complex medium. Community membership was largely determined by the donor feces, whereas relative abundances were determined by the supplemental carbon source. The absolute abundance of most taxa was independent of the supplementing nutrient, due to the ability of fast-growing organisms to quickly exhaust their niche in the complex medium and then exploit and monopolize the supplemental carbon source. Relative abundances of dominant taxa could be predicted from the nutritional preferences and growth dynamics of species in isolation, and exceptions were consistent with strain-level variation in growth capabilities. Our study reveals that community assembly follows simple rules of nutrient utilization dynamics and provides a predictive framework for manipulating gut commensal communities through nutritional perturbations.
]]></description>
<dc:creator>Aranda-Diaz, A.</dc:creator>
<dc:creator>Willis, L.</dc:creator>
<dc:creator>Nguyen, T. H.</dc:creator>
<dc:creator>Ho, P.-Y.</dc:creator>
<dc:creator>Vila, J.</dc:creator>
<dc:creator>Thomsen, T.</dc:creator>
<dc:creator>Chavez, T.</dc:creator>
<dc:creator>Yan, R.</dc:creator>
<dc:creator>Yu, F. B.</dc:creator>
<dc:creator>Neff, N.</dc:creator>
<dc:creator>Sanchez, A.</dc:creator>
<dc:creator>Estrela, S.</dc:creator>
<dc:creator>Huang, K.</dc:creator>
<dc:date>2023-01-14</dc:date>
<dc:identifier>doi:10.1101/2023.01.13.523996</dc:identifier>
<dc:title><![CDATA[Assembly of gut-derived bacterial communities follows "early-bird" resource utilization dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.13.523954v1?rss=1">
<title>
<![CDATA[
Next-generation interaction proteomics for quantitative Jumbophage-bacteria interaction mapping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.13.523954v1?rss=1</link>
<description><![CDATA[
Host-pathogen interactions (HPIs) are pivotal in regulating establishment, progression, and outcome of an infection. Affinity-purification mass spectrometry has become instrumental for the characterization of HPIs, however the targeted nature of exogenously expressing individual viral proteins has limited its utility to the analysis of relatively small pathogens. Here we present the use of co-fractionation mass spectrometry (SEC-MS) for the high-throughput analysis of HPIs from native viral infections of two jumbophages ({phi}KZ and{phi} PA3) in Pseudomonas aeruginosa. This enabled the detection >6000 unique host-pathogen and >200 pathogen-pathogen interactions for each phage, encompassing >50% of the phage proteome. Interactome-wide comparison across phages showed similar perturbed protein interactions suggesting fundamentally conserved mechanisms of phage predation within the KZ-like phage family. Prediction of novel ORFs revealed a{phi} PA3 complex showing strong structural and sequence similarity to{phi} KZ nvRNAp, suggesting{phi} PA3 also possesses two RNA polymerases acting at different stages of the infection cycle. We further expanded our understanding on the molecular organization of the virion packaged and injected proteome by identifying 23 novel virion components and 5 novel injected proteins, as well as providing the first evidence for interactions between KZ-like phage proteins and the host ribosome. To enable accessibility to this data, we developed PhageMAP, an online resource for network query, visualization, and interaction prediction (https://phagemap.ucsf.edu/). We anticipate this study will lay the foundation for the application of co-fractionation mass spectrometry for the scalable profiling of hostpathogen interactomes and protein complex dynamics upon infection.
]]></description>
<dc:creator>Fossati, A.</dc:creator>
<dc:creator>Mozumdar, D.</dc:creator>
<dc:creator>Kokontis, C. Y.</dc:creator>
<dc:creator>Nieweglowska, E. S.</dc:creator>
<dc:creator>Mendez, M.</dc:creator>
<dc:creator>Pelin, A.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Guo, B.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Agard, D.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:date>2023-01-15</dc:date>
<dc:identifier>doi:10.1101/2023.01.13.523954</dc:identifier>
<dc:title><![CDATA[Next-generation interaction proteomics for quantitative Jumbophage-bacteria interaction mapping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.13.524005v1?rss=1">
<title>
<![CDATA[
Functional microRNA-Targeting Drug Discovery by Graph-Based Deep Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.13.524005v1?rss=1</link>
<description><![CDATA[
MicroRNAs are recognized as key drivers in many cancers, but targeting them with small molecules remains a challenge. We present RiboStrike, a deep learning framework that identifies small molecules against specific microRNAs. To demonstrate its capabilities, we applied it to microRNA-21 (miR-21), a known driver of breast cancer. To ensure the selected molecules only targeted miR-21 and not other microRNAs, we also performed a counter-screen against DICER, an enzyme involved in microRNA biogenesis. Additionally, we used auxiliary models to evaluate toxicity and select the best candidates. Using datasets from various sources, we screened a pool of nine million molecules and identified eight, three of which showed anti-miR-21 activity in both reporter assays and RNA sequencing experiments. One of these was also tested in mouse models of breast cancer, resulting in a significant reduction of lung metastases. These results demonstrate RiboStrikes ability to effectively screen for microRNA-targeting compounds in cancer.
]]></description>
<dc:creator>Keshavarzi Arshadi, A.</dc:creator>
<dc:creator>Salem, M.</dc:creator>
<dc:creator>Karner, H.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Arab, A.</dc:creator>
<dc:creator>Yuan, J. S.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2023-01-16</dc:date>
<dc:identifier>doi:10.1101/2023.01.13.524005</dc:identifier>
<dc:title><![CDATA[Functional microRNA-Targeting Drug Discovery by Graph-Based Deep Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.16.524285v1?rss=1">
<title>
<![CDATA[
Arp2/3 Complex Activity Enables Nuclear YAP for Naive Pluripotency of Human Embryonic Stem Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.16.524285v1?rss=1</link>
<description><![CDATA[
Our understanding of transitions of human embryonic stem cells between distinct stages of pluripotency relies predominantly on regulation by transcriptional and epigenetic programs with limited insight on the role of established morphological changes. We report remodeling of the actin cytoskeleton of human embryonic stem cells (hESCs) as they transition from primed to naive pluripotency that includes assembly of a ring of contractile actin filaments encapsulating colonies of naive hESCs. Activity of the Arp2/3 complex is required for the actin ring, uniform cell mechanics within naive colonies, nuclear translocation of the Hippo pathway effectors YAP and TAZ, and effective transition to naive pluripotency. RNA-sequencing analysis confirms that Arp2/3 complex activity regulates Hippo signaling in hESCs, and impaired naive pluripotency with inhibited Arp2/3 complex activity is rescued by expressing a constitutively active, nuclear-localized YAP-S127A. These new findings on the cell biology of hESCs reveal a mechanism for cytoskeletal dynamics coordinating cell mechanics to regulate gene expression and facilitate transitions between pluripotency states.
]]></description>
<dc:creator>Meyer, N. P.</dc:creator>
<dc:creator>Singh, T.</dc:creator>
<dc:creator>Kutys, M. L.</dc:creator>
<dc:creator>Nystul, T.</dc:creator>
<dc:creator>Barber, D. L.</dc:creator>
<dc:date>2023-01-16</dc:date>
<dc:identifier>doi:10.1101/2023.01.16.524285</dc:identifier>
<dc:title><![CDATA[Arp2/3 Complex Activity Enables Nuclear YAP for Naive Pluripotency of Human Embryonic Stem Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.15.524101v1?rss=1">
<title>
<![CDATA[
The phospho-regulated amphiphysin/endophilin interaction is required for synaptic vesicle endocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.15.524101v1?rss=1</link>
<description><![CDATA[
The multidomain adaptor protein amphiphysin-1 (Amph1) is an important coordinator of clathrin-mediated endocytosis in non-neuronal cells and synaptic vesicle (SV) endocytosis at central nerve terminals. Amph1 contains a lipid-binding N-BAR (Bin/Amphiphysin/Rvs) domain, central proline-rich (PRD) and clathrin/AP2 (CLAP) domains, and a C-terminal SH3 domain. All domains interact with either lipids or SV endocytosis proteins, with all of these interactions required for SV endocytosis, apart from the Amph1 PRD. In this study, we determined this role and confirmed requirements for established Amph1 interactions in SV endocytosis at typical small central synapses. Domain-specific interactions of Amph1 were validated using in vitro GST pull-down assays, with the role of these interactions in SV endocytosis determined in molecular replacement experiments in primary neuronal culture. Using this approach, we confirmed important roles for CLAP and SH3 domain interactions in the control of SV endocytosis. Furthermore, we identified an interaction site for the endocytosis protein endophilin A1 in the Amph1 PRD and revealed a key role for this interaction in SV endocytosis. Finally, we discovered that the phosphorylation status of Amph1-S293 within the PRD dictates the formation of the Amph1-endophilin A1 complex and is essential for efficient SV regeneration. This work therefore identifies an activity-dependent dephosphorylation-dependent interaction that is key for efficient SV endocytosis.
]]></description>
<dc:creator>Kontaxi, C.</dc:creator>
<dc:creator>Cousin, M. A.</dc:creator>
<dc:date>2023-01-18</dc:date>
<dc:identifier>doi:10.1101/2023.01.15.524101</dc:identifier>
<dc:title><![CDATA[The phospho-regulated amphiphysin/endophilin interaction is required for synaptic vesicle endocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.17.523348v1?rss=1">
<title>
<![CDATA[
Normative modeling of brain morphometry in Clinical High-Risk for Psychosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.17.523348v1?rss=1</link>
<description><![CDATA[
ImportanceThe lack of robust neuroanatomical markers of psychosis risk has been traditionally attributed to heterogeneity. A complementary hypothesis is that variation in neuroanatomical measures in the majority of individuals at psychosis risk may be nested within the range observed in healthy individuals.

ObjectiveTo quantify deviations from the normative range of neuroanatomical variation in individuals at clinical high-risk for psychosis (CHR-P) and evaluate their overlap with healthy variation and their association with positive symptoms, cognition, and conversion to a psychotic disorder.

Design, Setting, and ParticipantsClinical, IQ and FreeSurfer-derived regional measures of cortical thickness (CT), cortical surface area (SA), and subcortical volume (SV) from 1,340 CHR-P individuals [47.09% female; mean age: 20.75 (4.74) years] and 1,237 healthy individuals [44.70% female; mean age: 22.32 (4.95) years] from 29 international sites participating in the ENIGMA Clinical High Risk for Psychosis Working Group.

Main Outcomes and MeasuresFor each regional morphometric measure, z-scores were computed that index the degree of deviation from the normative means of that measure in a healthy reference population (N=37,407). Average deviation scores (ADS) for CT, SA, SV, and globally across all measures (G) were generated by averaging the respective regional z-scores. Regression analyses were used to quantify the association of deviation scores with clinical severity and cognition and two-proportion z-tests to identify case-control differences in the proportion of individuals with infranormal (z<-1.96) or supranormal (z>1.96) scores.

ResultsCHR-P and healthy individuals overlapped in the distributions of the observed values, regional z-scores, and all ADS vales. The proportion of CHR-P individuals with infranormal or supranormal values in any metric was low (<12%) and similar to that of healthy individuals. CHR-P individuals who converted to psychosis compared to those who did not convert had a higher percentage of infranormal values in temporal regions (5-7% vs 0.9-1.4%). In the CHR-P group, only the ADSSA showed significant but weak associations (|{beta}|<0.09; PFDR<0.05) with positive symptoms and IQ.

Conclusions and RelevanceThe study findings challenge the usefulness of macroscale neuromorphometric measures as diagnostic biomarkers of psychosis risk and suggest that such measures do not provide an adequate explanation for psychosis risk.

Key pointsO_ST_ABSQuestionC_ST_ABSIs the risk of psychosis associated with brain morphometric changes that deviate significantly from healthy variation?

FindingsIn this study of 1340 individuals high-risk for psychosis (CHR-P) and 1237 healthy participants, individual-level variation in macroscale neuromorphometric measures of the CHR-P group was largely nested within healthy variation and was not associated with the severity of positive psychotic symptoms or conversion to a psychotic disorder.

MeaningThe findings suggest the macroscale neuromorphometric measures have limited utility as diagnostic biomarkers of psychosis risk.
]]></description>
<dc:creator>Haas, S. S.</dc:creator>
<dc:creator>Ge, R.</dc:creator>
<dc:creator>Agartz, I.</dc:creator>
<dc:creator>Amminger, G. P.</dc:creator>
<dc:creator>Andreassen, O. A.</dc:creator>
<dc:creator>Bachman, P.</dc:creator>
<dc:creator>Baeza, I.</dc:creator>
<dc:creator>Choi, S.</dc:creator>
<dc:creator>Colibazzi, T.</dc:creator>
<dc:creator>Cropley, V. L.</dc:creator>
<dc:creator>de la Fuente-Sandoval, C.</dc:creator>
<dc:creator>Ebdrup, B. H.</dc:creator>
<dc:creator>Fortea, A.</dc:creator>
<dc:creator>Fusar-Poli, P.</dc:creator>
<dc:creator>Glenthoj, B. Y.</dc:creator>
<dc:creator>Glenthoj, L. B.</dc:creator>
<dc:creator>Haut, K. M.</dc:creator>
<dc:creator>Hayes, R. A.</dc:creator>
<dc:creator>Heekeren, K.</dc:creator>
<dc:creator>Hooker, C. I.</dc:creator>
<dc:creator>Hwang, W. J.</dc:creator>
<dc:creator>Jahanshad, N.</dc:creator>
<dc:creator>Kaess, M.</dc:creator>
<dc:creator>Kasai, K.</dc:creator>
<dc:creator>Katagiri, N.</dc:creator>
<dc:creator>Kim, M.</dc:creator>
<dc:creator>Kindler, J.</dc:creator>
<dc:creator>Koike, S.</dc:creator>
<dc:creator>Kristensen, T. D.</dc:creator>
<dc:creator>Kwon, J. S.</dc:creator>
<dc:creator>Lawrie, S. M.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Lemmers-Jansen, I. L.</dc:creator>
<dc:creator>Lin, A.</dc:creator>
<dc:creator>Ma, X.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>McGuire, P.</dc:creator>
<dc:creator>Michel, C.</dc:creator>
<dc:creator>Mizrahi, R.</dc:creator>
<dc:creator>Mizuno, M.</dc:creator>
<dc:creator>Moller, P.</dc:creator>
<dc:creator>Mora-Duran, R.</dc:creator>
<dc:creator>Nelson, B.</dc:creator>
<dc:creator>Nemot</dc:creator>
<dc:date>2023-01-18</dc:date>
<dc:identifier>doi:10.1101/2023.01.17.523348</dc:identifier>
<dc:title><![CDATA[Normative modeling of brain morphometry in Clinical High-Risk for Psychosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.16.523256v1?rss=1">
<title>
<![CDATA[
Extensive and Persistent Extravascular Dermal Fibrin Deposition Characterizes Systemic Sclerosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.16.523256v1?rss=1</link>
<description><![CDATA[
Systemic sclerosis (SSc) is an autoimmune disease characterized by progressive multiorgan fibrosis. While the cause of SSc remains unknown, a perturbed vasculature is considered a critical early step in the pathogenesis. Using fibrinogen as a marker of vascular leakage, we found extensive extravascular fibrinogen deposition in the dermis of both limited and diffuse systemic sclerosis disease, and it was present in both early and late-stage patients. Based on a timed series of excision wounds, retention on the fibrin deposit of the splice variant domain, fibrinogen EC, indicated a recent event, while fibrin networks lacking the EC domain were older. Application of this timing tool to SSc revealed considerable heterogeneity in EC domain distribution providing unique insight into disease activity. Intriguingly, the fibrinogen-EC domain also accumulated in macrophages. These observations indicate that systemic sclerosis is characterized by ongoing vascular leakage resulting in extensive interstitial fibrin deposition that is either continually replenished and/or there is impaired fibrin clearance. Unresolved fibrin deposition might then incite chronic tissue remodeling.
]]></description>
<dc:creator>Browning, J. L.</dc:creator>
<dc:creator>Bhawan, J.</dc:creator>
<dc:creator>Tseng, A.</dc:creator>
<dc:creator>Crossland, N.</dc:creator>
<dc:creator>Bujor, A. M.</dc:creator>
<dc:creator>Akassoglou, K.</dc:creator>
<dc:creator>Assassi, S.</dc:creator>
<dc:creator>Skaug, B.</dc:creator>
<dc:creator>Ho, J.</dc:creator>
<dc:date>2023-01-19</dc:date>
<dc:identifier>doi:10.1101/2023.01.16.523256</dc:identifier>
<dc:title><![CDATA[Extensive and Persistent Extravascular Dermal Fibrin Deposition Characterizes Systemic Sclerosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.17.524447v1?rss=1">
<title>
<![CDATA[
Molecular Logic of Synaptic Diversity Between Drosophila Tonic and Phasic Motoneurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.17.524447v1?rss=1</link>
<description><![CDATA[
Although neuronal subtypes display unique synaptic organization and function, the underlying transcriptional differences that establish these features is poorly understood. To identify molecular pathways that contribute to synaptic diversity, single neuron PatchSeq RNA profiling was performed on Drosophila tonic and phasic glutamatergic motoneurons. Tonic motoneurons form weaker facilitating synapses onto single muscles, while phasic motoneurons form stronger depressing synapses onto multiple muscles. Super-resolution microscopy and in vivo imaging demonstrated synaptic active zones in phasic motoneurons are more compact and display enhanced Ca2+ influx compared to their tonic counterparts. Genetic analysis identified unique synaptic properties that mapped onto gene expression differences for several cellular pathways, including distinct signaling ligands, post-translational modifications and intracellular Ca2+ buffers. These findings provide insights into how unique transcriptomes drive functional and morphological differences between neuronal subtypes.
]]></description>
<dc:creator>Jetti, S. J.</dc:creator>
<dc:creator>Crane, A. B.</dc:creator>
<dc:creator>Akbergenova, Y.</dc:creator>
<dc:creator>Aponte-Santiago, N. A.</dc:creator>
<dc:creator>Cunningham, K. L.</dc:creator>
<dc:creator>Whittaker, C. W.</dc:creator>
<dc:creator>Littleton, J. T.</dc:creator>
<dc:date>2023-01-19</dc:date>
<dc:identifier>doi:10.1101/2023.01.17.524447</dc:identifier>
<dc:title><![CDATA[Molecular Logic of Synaptic Diversity Between Drosophila Tonic and Phasic Motoneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.18.524601v1?rss=1">
<title>
<![CDATA[
Human Bone Marrow Plasma Cell Atlas: Maturation and Survival Pathways Unraveled by Single Cell Analyses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.18.524601v1?rss=1</link>
<description><![CDATA[
Human bone marrow (BM) plasma cells are heterogeneous, ranging from newly arrived antibody-secreting cells (ASC) to long-lived plasma cells (LLPC). We provide single cell transcriptional resolution of 17,347 BM ASC from 5 healthy adults. Fifteen clusters were identified ranging from newly minted ASC (cluster 1) expressing MKI67 and high MHC Class II that progressed to late clusters 5-8 through intermediate clusters 2-4. Additional clusters included early and late IgM-predominant ASC of likely extra-follicular origin; IFN-responsive; and high mitochondrial activity ASC. Late ASCs were distinguished by differences in G2M checkpoints, MTOR signaling, distinct metabolic pathways, CD38 expression, and utilization of TNF-receptor superfamily members. They mature through two distinct paths differentiated by the degree of TNF signaling through NFKB. This study provides the first single cell resolution atlas and molecular roadmap of LLPC maturation, thereby providing insight into differentiation trajectories and molecular regulation of these essential processes in the human BM microniche. This information enables investigation of the origin of protective and pathogenic antibodies in multiple diseases and development of new strategies targeted to the enhancement or depletion of the corresponding ASC.

One Sentence Summary: The single cell transcriptomic atlas of human bone marrow plasma cell heterogeneity shows maturation of class-switched early and late subsets, specific IgM and Interferon-driven clusters, and unique heterogeneity of the late subsets which encompass the long-lived plasma cells.
]]></description>
<dc:creator>Duan, M.</dc:creator>
<dc:creator>Nguyen, D. C.</dc:creator>
<dc:creator>Joyner, C. J.</dc:creator>
<dc:creator>Saney, C.</dc:creator>
<dc:creator>Tipton, C.</dc:creator>
<dc:creator>Andrews, J.</dc:creator>
<dc:creator>Lonial, S.</dc:creator>
<dc:creator>Kim, C.</dc:creator>
<dc:creator>Hentenaar, I.</dc:creator>
<dc:creator>Kosters, A.</dc:creator>
<dc:creator>Ghosn, E.</dc:creator>
<dc:creator>Jackson, A.</dc:creator>
<dc:creator>Knechtle, S.</dc:creator>
<dc:creator>Maruthamuthu, S.</dc:creator>
<dc:creator>Chandran, S.</dc:creator>
<dc:creator>Martin, T.</dc:creator>
<dc:creator>Rajalingam, R.</dc:creator>
<dc:creator>Vincenti, F.</dc:creator>
<dc:creator>Breeden, C.</dc:creator>
<dc:creator>Sanz, I.</dc:creator>
<dc:creator>Gibson, G.</dc:creator>
<dc:creator>Lee, F. E.-H.</dc:creator>
<dc:date>2023-01-20</dc:date>
<dc:identifier>doi:10.1101/2023.01.18.524601</dc:identifier>
<dc:title><![CDATA[Human Bone Marrow Plasma Cell Atlas: Maturation and Survival Pathways Unraveled by Single Cell Analyses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.18.524608v1?rss=1">
<title>
<![CDATA[
Translational predictions of phase 2a first-in patient efficacy studies for antituberculosis drugs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.18.524608v1?rss=1</link>
<description><![CDATA[
BackgroundPhase 2a trials in tuberculosis typically use early bactericidal activity (EBA), the decline in sputum colony forming units (CFU) over 14 days, as the primary outcome for testing the efficacy of drugs as monotherapy. However, the cost of phase 2a trials can range from 7 to 19.6 million dollars on average, while more than 30% of drugs fail to progress to phase 3. Better utilizing preclinical data to predict and prioritize the most likely drugs to succeed will thus help accelerate drug development and reduce costs. We aim to predict clinical EBA using preclinical in vivo pharmacokinetic-pharmacodynamic (PKPD) data and a model-based translational pharmacology approach.

Methods and FindingsFirst, mouse PK, PD and clinical PK models were compiled. Second, mouse PKPD models were built to derive an exposure response relationship. Third, translational prediction of clinical EBA studies was performed using mouse PKPD relationships and informed by clinical PK models and species-specific protein binding. Presence or absence of clinical efficacy was accurately predicted from the mouse model. Predicted daily decreases of CFU in the first 2 days of treatment and between day 2 and day 14 were consistent with clinical observations.

ConclusionThis platform provides an innovative solution to inform or even replace phase 2a EBA trials, to bridge the gap between mouse efficacy studies and phase 2b and phase 3 trials, and to substantially accelerate drug development.
]]></description>
<dc:creator>Savic, R. M.</dc:creator>
<dc:creator>Goh, J. J. N.</dc:creator>
<dc:creator>Ernest, J. P.</dc:creator>
<dc:creator>Strydom, N.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>van Wijk, R. C.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Deitchman, A.</dc:creator>
<dc:creator>Nuermberger, E.</dc:creator>
<dc:date>2023-01-20</dc:date>
<dc:identifier>doi:10.1101/2023.01.18.524608</dc:identifier>
<dc:title><![CDATA[Translational predictions of phase 2a first-in patient efficacy studies for antituberculosis drugs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.18.524611v1?rss=1">
<title>
<![CDATA[
Frontotemporal Dementia Patient Neurons With Progranulin Deficiency Display Protein Dyshomeostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.18.524611v1?rss=1</link>
<description><![CDATA[
Haploinsufficiency of progranulin (PGRN) causes frontotemporal dementia (FTD), a devastating neurodegenerative disease with no effective treatment. PGRN is required for efficient proteostasis, as loss of neuronal PGRN results in dysfunctional lysosomes and impaired clearance and cytoplasmic aggregation of TDP-43, a protein involved in neurodegeneration in FTD. These and other events lead to neurodegeneration and neuroinflammation. However, the detailed mechanisms leading to protein dyshomeostasis in PGRN-deficient cells remain unclear. We report here the development of human cell models of FTD with PGRN-deficiency to explore the molecular mechanisms underlying proteostasis breakdown and TDP-43 aggregation in FTD. Neurons differentiated from FTD patient induced pluripotent stem cells (iPSCs) have reduced PGRN levels, and the neurons recapitulate key disease features, including impaired lysosomal function, defective TDP-43 turnover and accumulation, neurodegeneration, and death. Proteomic analysis revealed altered levels of proteins linked to the autophagy-lysosome pathway (ALP) and the ubiquitin-proteasome system (UPS) in FTD patient neurons, providing new mechanistic insights into the link between PGRN-deficiency and disease pathobiology.
]]></description>
<dc:creator>Elia, L. P.</dc:creator>
<dc:creator>Herting, B.</dc:creator>
<dc:creator>Alijagic, A.</dc:creator>
<dc:creator>Buselli, C.</dc:creator>
<dc:creator>Wong, L.</dc:creator>
<dc:creator>Morrison, G.</dc:creator>
<dc:creator>Prado, M. A.</dc:creator>
<dc:creator>Paolo, J. A.</dc:creator>
<dc:creator>Gygi, S.</dc:creator>
<dc:creator>Finley, D.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:date>2023-01-20</dc:date>
<dc:identifier>doi:10.1101/2023.01.18.524611</dc:identifier>
<dc:title><![CDATA[Frontotemporal Dementia Patient Neurons With Progranulin Deficiency Display Protein Dyshomeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.19.524562v1?rss=1">
<title>
<![CDATA[
Development and characterization of new patient-derived xenograft (PDX) models of osteosarcoma with distinct metastatic capacities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.19.524562v1?rss=1</link>
<description><![CDATA[
Models to study metastatic disease in rare cancers are needed to advance preclinical therapeutics and to gain insight into disease biology, especially for highly aggressive cancers with a propensity for metastatic spread. Osteosarcoma is a rare cancer with a complex genomic landscape in which outcomes for patients with metastatic disease are poor. As osteosarcoma genomes are highly heterogeneous, a large panel of models is needed to fully elucidate key aspects of disease biology and to recapitulate clinically-relevant phenotypes. We describe the development and characterization of osteosarcoma patient-derived xenografts (PDXs) and a panel of PDX-derived cell lines. Matched patient samples, PDXs, and PDX-derived cell lines were comprehensively evaluated using whole genome sequencing and RNA sequencing. PDXs and PDX-derived cell lines largely maintained the expression profiles of the patient from which they were derived despite the emergence of whole-genome duplication (WGD) in a subset of cell lines. These cell line models were heterogeneous in their metastatic capacity and their tissue tropism as observed in both intravenous and orthotopic models. As proof-of-concept study, we used one of these models to test the preclinical effectiveness of a CDK inhibitor on the growth of metastatic tumors in an orthotopic amputation model. Single-agent dinaciclib was effective at dramatically reducing the metastatic burden in this model.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=74 SRC="FIGDIR/small/524562v1_ufig1.gif" ALT="Figure 1">
View larger version (32K):
org.highwire.dtl.DTLVardef@17ea2a8org.highwire.dtl.DTLVardef@e8fcc7org.highwire.dtl.DTLVardef@2b9f8corg.highwire.dtl.DTLVardef@1f7b802_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Schott, C. R.</dc:creator>
<dc:creator>Koehne, A. L.</dc:creator>
<dc:creator>Sayles, L. C.</dc:creator>
<dc:creator>Young, E. P.</dc:creator>
<dc:creator>Luck, C.</dc:creator>
<dc:creator>Yu, K.</dc:creator>
<dc:creator>Lee, A. G.</dc:creator>
<dc:creator>Breese, M. R.</dc:creator>
<dc:creator>Leung, S. G.</dc:creator>
<dc:creator>Shah, A. T.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Liu, H.-Y.</dc:creator>
<dc:creator>Spillinger, A.</dc:creator>
<dc:creator>Behroozfard, I. H.</dc:creator>
<dc:creator>Marini, K. D.</dc:creator>
<dc:creator>Dinh, P. T.</dc:creator>
<dc:creator>Pons Ventura, M. V.</dc:creator>
<dc:creator>Vanderboon, E. N.</dc:creator>
<dc:creator>Hazard, F. K.</dc:creator>
<dc:creator>Cho, S.-J.</dc:creator>
<dc:creator>Avedian, R. S.</dc:creator>
<dc:creator>Mohler, D. G.</dc:creator>
<dc:creator>Zimel, M.</dc:creator>
<dc:creator>Wustrack, R.</dc:creator>
<dc:creator>Curtis, C.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Sweet-Cordero, E. A.</dc:creator>
<dc:date>2023-01-20</dc:date>
<dc:identifier>doi:10.1101/2023.01.19.524562</dc:identifier>
<dc:title><![CDATA[Development and characterization of new patient-derived xenograft (PDX) models of osteosarcoma with distinct metastatic capacities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.19.524758v1?rss=1">
<title>
<![CDATA[
Mycobacterium tuberculosis resides in lysosome-poor monocyte-derived lung cells during persistent infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.19.524758v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) infects cells in multiple lung myeloid cell subsets and causes chronic infection despite innate and adaptive immune responses. However, the mechanisms allowing Mtb to evade elimination are not fully understood. Here, using new methods, we determined that after T cell responses have developed, CD11clo monocyte-derived lung cells termed MNC1 (mononuclear cell subset 1), harbor more live Mtb compared to alveolar macrophages (AM), neutrophils, and less permissive CD11chi MNC2. Bulk RNA sequencing of sorted cells revealed that the lysosome biogenesis pathway is underexpressed in MNC1. Functional assays confirmed that Mtb-permissive MNC1 have less lysosome content, acidification, and proteolytic activity than AM, and less nuclear TFEB, a master regulator of lysosome biogenesis. Mtb infection does not drive lysosome deficiency in MNC1 in vivo. Instead, Mtb recruits MNC1 and MNC2 to the lungs for its spread from AM to these cell subsets as a virulence mechanism that requires the Mtb ESX-1 secretion system. The c-Abl tyrosine kinase inhibitor nilotinib activates TFEB and enhances lysosome function of primary macrophages in vitro and MNC1 and MNC2 in vivo, improving control of Mtb infection. Our results indicate that Mtb exploits lysosome-poor monocyte-derived cells for in vivo persistence, suggesting a potential target for host-directed tuberculosis therapy.

One Sentence SummaryVirulent Mtb recruits and exploits intrinsically lysosome-deficient lung mononuclear cell subsets to resist elimination during chronic infection.
]]></description>
<dc:creator>Zheng, W.</dc:creator>
<dc:creator>Chang, I.-C.</dc:creator>
<dc:creator>Limberis, J.</dc:creator>
<dc:creator>Budzik, J. M.</dc:creator>
<dc:creator>Zha, B. S.</dc:creator>
<dc:creator>Howard, Z.</dc:creator>
<dc:creator>Ernst, J. D.</dc:creator>
<dc:date>2023-01-20</dc:date>
<dc:identifier>doi:10.1101/2023.01.19.524758</dc:identifier>
<dc:title><![CDATA[Mycobacterium tuberculosis resides in lysosome-poor monocyte-derived lung cells during persistent infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.20.524596v1?rss=1">
<title>
<![CDATA[
Dopamine D2 receptors in nucleus accumbens cholinergic interneurons increase impulsive choice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.20.524596v1?rss=1</link>
<description><![CDATA[
Impulsive choice, often characterized by excessive preference for small, short-term rewards over larger, long-term rewards, is a prominent feature of substance use and other neuropsychiatric disorders. The neural mechanisms underlying impulsive choice are not well understood, but growing evidence implicates nucleus accumbens (NAc) dopamine and its actions on dopamine D2 receptors (D2Rs). Because several NAc cell types and afferents express D2Rs, it has been difficult to determine the specific neural mechanisms linking NAc D2Rs to impulsive choice. Of these cell types, cholinergic interneurons (CINs) of the NAc, which express D2Rs, have emerged as key regulators of striatal output and local dopamine release. Despite these relevant functions, whether D2Rs expressed specifically in these neurons contribute to impulsive choice behavior is unknown. Here, we show that D2R upregulation in CINs of the mouse NAc increases impulsive choice as measured in a delay discounting task without affecting reward magnitude sensitivity or interval timing. Conversely, mice lacking D2Rs in CINs showed decreased delay discounting. Furthermore, CIN D2R manipulations did not affect probabilistic discounting, which measures a different form of impulsive choice. Together, these findings suggest that CIN D2Rs regulate impulsive decision-making involving delay costs, providing new insight into the mechanisms by which NAc dopamine influences impulsive behavior.
]]></description>
<dc:creator>Cavallaro, J.</dc:creator>
<dc:creator>Yeisley, J.</dc:creator>
<dc:creator>Akdogan, B.</dc:creator>
<dc:creator>Floeder, J. R.</dc:creator>
<dc:creator>Balsam, P. D.</dc:creator>
<dc:creator>Gallo, E. F.</dc:creator>
<dc:date>2023-01-20</dc:date>
<dc:identifier>doi:10.1101/2023.01.20.524596</dc:identifier>
<dc:title><![CDATA[Dopamine D2 receptors in nucleus accumbens cholinergic interneurons increase impulsive choice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.20.524922v1?rss=1">
<title>
<![CDATA[
Lipopolysaccharide integrity primes bacterial sensitivity to a cell wall-degrading intermicrobial toxin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.20.524922v1?rss=1</link>
<description><![CDATA[
Gram-negative bacteria can antagonize neighboring microbes using a type VI secretion system (T6SS) to deliver toxins that target different essential cellular features. Despite the conserved nature of these targets, T6SS potency can vary across recipient species. To understand the molecular basis of intrinsic T6SS susceptibility, we screened for essential Escherichia coli genes that affect its survival when antagonized by a cell wall-degrading T6SS toxin from Pseudomonas aeruginosa, Tae1. We revealed genes associated with both the cell wall and a separate layer of the cell envelope, surface lipopolysaccharide, that modulate Tae1 toxicity in vivo. Disruption of lipopolysaccharide synthesis provided Escherichia coli (Eco) with novel resistance to Tae1, despite significant cell wall degradation. These data suggest that Tae1 toxicity is determined not only by direct substrate damage, but also by indirect cell envelope homeostasis activities. We also found that Tae1-resistant Eco exhibited reduced cell wall synthesis and overall slowed growth, suggesting that reactive cell envelope maintenance pathways could promote, not prevent, self-lysis. Together, our study highlights the consequences of co-regulating essential pathways on recipient fitness during interbacterial competition, and how antibacterial toxins leverage cellular vulnerabilities that are both direct and indirect to their specific targets in vivo.
]]></description>
<dc:creator>Trotta, K. L.</dc:creator>
<dc:creator>Hayes, B.</dc:creator>
<dc:creator>Schneider, J. P.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Todor, H.</dc:creator>
<dc:creator>Grimes, P. R.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Hatleberg, W.</dc:creator>
<dc:creator>Silvis, M.</dc:creator>
<dc:creator>Kim, R.</dc:creator>
<dc:creator>Koo, B.-M.</dc:creator>
<dc:creator>Basler, M.</dc:creator>
<dc:creator>Chou, S.</dc:creator>
<dc:date>2023-01-21</dc:date>
<dc:identifier>doi:10.1101/2023.01.20.524922</dc:identifier>
<dc:title><![CDATA[Lipopolysaccharide integrity primes bacterial sensitivity to a cell wall-degrading intermicrobial toxin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.21.525036v1?rss=1">
<title>
<![CDATA[
Single-cell and spatial multi-omics identify innate and stromal modules targeted by anti-integrin therapy in ulcerative colitis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.21.525036v1?rss=1</link>
<description><![CDATA[
Ulcerative colitis (UC) is driven by immune and stromal subsets, culminating in epithelial injury. Vedolizumab (VDZ) is an anti-integrin antibody that is effective for treating UC. VDZ is known to inhibit lymphocyte trafficking to the intestine, but its broader effects on other cell subsets are less defined. To identify the inflammatory cells that contribute to colitis and are affected by VDZ, we performed single-cell transcriptomic and proteomic analyses of peripheral blood and colonic biopsies in healthy controls and patients with UC on VDZ or other therapies. Here we show that VDZ treatment is associated with alterations in circulating and tissue mononuclear phagocyte (MNP) subsets, along with modest shifts in lymphocytes. Spatial multi-omics of formalin-fixed biopsies demonstrates trends towards increased abundance and proximity of MNP and fibroblast subsets in active colitis. Spatial transcriptomics of archived specimens pre-treatment identifies epithelial-, MNP-, and fibroblast-enriched genes related to VDZ responsiveness, highlighting important roles for these subsets in UC.
]]></description>
<dc:creator>Mennillo, E.</dc:creator>
<dc:creator>Kim, Y. J.</dc:creator>
<dc:creator>Rusu, I.</dc:creator>
<dc:creator>Lee, G.</dc:creator>
<dc:creator>Dorman, L. C.</dc:creator>
<dc:creator>Bernard-Vazquez, F.</dc:creator>
<dc:creator>Bain, J. L.</dc:creator>
<dc:creator>Patel, R.</dc:creator>
<dc:creator>Andersen, C.</dc:creator>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Tamaki, S.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Shen, A.</dc:creator>
<dc:creator>Naser, M.</dc:creator>
<dc:creator>Eckalbar, W.</dc:creator>
<dc:creator>Cho, S.-j.</dc:creator>
<dc:creator>Beck, K.</dc:creator>
<dc:creator>El-Nachef, N.</dc:creator>
<dc:creator>Lewin, S.</dc:creator>
<dc:creator>Selvig, D. R.</dc:creator>
<dc:creator>Terdiman, J. P.</dc:creator>
<dc:creator>Mahadevan, U.</dc:creator>
<dc:creator>Oh, D.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Pisco, A.</dc:creator>
<dc:creator>Combes, A.</dc:creator>
<dc:creator>Kattah, M. G.</dc:creator>
<dc:date>2023-01-22</dc:date>
<dc:identifier>doi:10.1101/2023.01.21.525036</dc:identifier>
<dc:title><![CDATA[Single-cell and spatial multi-omics identify innate and stromal modules targeted by anti-integrin therapy in ulcerative colitis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.22.524916v1?rss=1">
<title>
<![CDATA[
Supporting central nervous system neuroprotection and remyelination by specific TLR4 antagonism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.22.524916v1?rss=1</link>
<description><![CDATA[
ApTOLL is an aptamer specifically designed to antagonize Toll-Like Receptor 4 (TLR4), a relevant actor for innate immunity involved in inflammatory responses in multiple sclerosis (MS) and other diseases. MS is a primary demyelinating, chronic, inmune and neurodegenerative disease of the central nervous system that normally debuts in young adults. The currently available therapeutic arsenal to treat MS is composed of immunomodulators but, to date, there are no (re)myelinating drugs available in clinics. Our present study shows cells expressing TLR4 in demyelinating lesions of MS patients (postmortem samples from cerebral cortex) and, as a derivative, we studied the effect of TLR4 inhibition with ApTOLL in animal models of MS (experimental autoimmune encephalomyelitis -EAE- and the cuprizone). The treatment with ApTOLL positively impacted the clinical symptomatology, and this was associated with better preservation plus restoration of myelin and oligodendrocytes in the demyelinated lesions of these animals, which suggests not only an immunomodulatory but also a remyelinating effect of the treatment with ApTOLL. This latter was corroborated on purified cultures of rodent and adult human oligodendrocyte precursor cells (OPCs), confirming the expression of TLR4 in this cell type. Altogether, the molecular nature of ApTOLL and its mechanism/s of action strongly supports this compound as a novel candidate to treat MS and other demyelinating scenarios.
]]></description>
<dc:creator>Fernandez-Gomez, B.</dc:creator>
<dc:creator>Marchena, M. A.</dc:creator>
<dc:creator>Pineiro, D.</dc:creator>
<dc:creator>Lao, Y.</dc:creator>
<dc:creator>Valencia, G.</dc:creator>
<dc:creator>Nocera, S.</dc:creator>
<dc:creator>Benitez-Fernandez, R.</dc:creator>
<dc:creator>Gomez-Martin, P.</dc:creator>
<dc:creator>Castano-Leon, A. M.</dc:creator>
<dc:creator>Lagares, A.</dc:creator>
<dc:creator>Hernandez-Jimenez, M.</dc:creator>
<dc:creator>de Castro, F.</dc:creator>
<dc:date>2023-01-22</dc:date>
<dc:identifier>doi:10.1101/2023.01.22.524916</dc:identifier>
<dc:title><![CDATA[Supporting central nervous system neuroprotection and remyelination by specific TLR4 antagonism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.22.525086v1?rss=1">
<title>
<![CDATA[
A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.22.525086v1?rss=1</link>
<description><![CDATA[
Mapping genetic interactions is essential for determining gene function and defining novel biological pathways. We report a simple to use CRISPR interference (CRISPRi) based platform, compatible with Fluorescence Activated Cell Sorting (FACS)-based reporter screens, to query epistatic relationships at scale. This is enabled by a flexible dual-sgRNA library design that allows for the simultaneous delivery and selection of a fixed sgRNA and a second randomized guide, comprised of a genome-wide library, with a single transduction. We use this approach to identify epistatic relationships for a defined biological pathway, showing both increased sensitivity and specificity than traditional growth screening approaches.
]]></description>
<dc:creator>Guna, A.</dc:creator>
<dc:creator>Page, K. R.</dc:creator>
<dc:creator>Replogle, J. R.</dc:creator>
<dc:creator>Esantsi, T. K.</dc:creator>
<dc:creator>Wang, M. L.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:creator>Voorhees, R. M.</dc:creator>
<dc:date>2023-01-22</dc:date>
<dc:identifier>doi:10.1101/2023.01.22.525086</dc:identifier>
<dc:title><![CDATA[A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.23.524428v1?rss=1">
<title>
<![CDATA[
Notch1 cortical signaling regulates epithelial architecture and cell-cell adhesion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.23.524428v1?rss=1</link>
<description><![CDATA[
Notch receptors control tissue morphogenic processes that involve coordinated changes in cell architecture and gene expression, but how a single receptor can produce these diverse biological outputs is unclear. Here we employ a 3D organotypic model of a ductal epithelium to reveal tissue morphogenic defects result from loss of Notch1, but not Notch1 transcriptional signaling. Instead, defects in duct morphogenesis are driven by dysregulated epithelial cell architecture and mitogenic signaling which result from loss of a transcription-independent Notch1 cortical signaling mechanism that ultimately functions to stabilize adherens junctions and cortical actin. We identify that Notch1 localization and cortical signaling are tied to apical-basal cell restructuring and discover a Notch1-FAM83H interaction underlies stabilization of adherens junctions and cortical actin. Together, these results offer new insights into Notch1 signaling and regulation, and advance a paradigm in which transcriptional and cell adhesive programs might be coordinated by a single receptor.
]]></description>
<dc:creator>White, M. J.</dc:creator>
<dc:creator>Jacobs, K. A.</dc:creator>
<dc:creator>Singh, T.</dc:creator>
<dc:creator>Kutys, M. L.</dc:creator>
<dc:date>2023-01-23</dc:date>
<dc:identifier>doi:10.1101/2023.01.23.524428</dc:identifier>
<dc:title><![CDATA[Notch1 cortical signaling regulates epithelial architecture and cell-cell adhesion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.23.525156v1?rss=1">
<title>
<![CDATA[
Insights into the mechanism of oligodendrocyte protection and remyelination enhancement by the integrated stress response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.23.525156v1?rss=1</link>
<description><![CDATA[
CNS inflammation triggers activation of the integrated stress response (ISR). We previously reported that prolonging the ISR protects remyelinating oligodendrocytes and promotes remyelination in the presence of inflammation (Chen et al., eLife, 2021). However, the exact mechanisms through which this occurs remain unknown. Here, we investigated whether the ISR modulator Sephin1 in combination with the oligodendrocyte differentiation enhancing reagent bazedoxifene (BZA) is able to accelerate remyelination under inflammation, and the underlying mechanisms mediating this pathway. We find that the combined treatment of Sephin1 and BZA is sufficient to accelerate early-stage remyelination in mice with ectopic IFN-{gamma} expression in the CNS. IFN-{gamma}, which is a critical inflammatory cytokine in multiple sclerosis (MS), inhibits oligodendrocyte precursor cell (OPC) differentiation in culture and triggers a mild ISR. Mechanistically, we further show that BZA promotes OPC differentiation in the presence of IFN-{gamma}, while Sephin1 enhances the IFN-{gamma}-induced ISR by reducing protein synthesis and increasing RNA stress granule formation in differentiating oligodendrocytes. Finally, the ISR suppressor 2BAct is able to partially lessen the beneficial effect of Sephin1 on disease progression, in an MS mouse model of experimental autoimmune encephalitis (EAE). Overall, our findings uncover distinct mechanisms of action of BZA and Sephin1 on oligodendrocyte lineage cells under inflammatory stress, suggesting that a combination therapy may effectively promote restoring neuronal function in MS patients.
]]></description>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Quan, S.</dc:creator>
<dc:creator>Patil, V.</dc:creator>
<dc:creator>Kunjamma, R. B.</dc:creator>
<dc:creator>Tokars, H. M.</dc:creator>
<dc:creator>Leisten, E. D.</dc:creator>
<dc:creator>Chan, J. R.</dc:creator>
<dc:creator>Wong, Y.</dc:creator>
<dc:creator>Popko, B. J.</dc:creator>
<dc:date>2023-01-23</dc:date>
<dc:identifier>doi:10.1101/2023.01.23.525156</dc:identifier>
<dc:title><![CDATA[Insights into the mechanism of oligodendrocyte protection and remyelination enhancement by the integrated stress response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.23.525251v1?rss=1">
<title>
<![CDATA[
Conformationally Adaptive Dyes Enable Protein-Adaptive Differential Scanning Fluorometry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.23.525251v1?rss=1</link>
<description><![CDATA[
Flexible in vitro methods alter the course of biological discoveries. Differential Scanning Fluorimetry (DSF) is a particularly versatile technique which reports protein thermal unfolding via fluorogenic dye. However, applications of DSF are limited by widespread protein incompatibilities with the available DSF dyes. Here, we enable DSF applications for 66 of 70 tested proteins (94%) including 10 from the SARS-CoV2 virus using a chemically diverse dye library, Aurora, to identify compatible dye-protein pairs in high throughput. We find that this protein-adaptive DSF platform (paDSF) not only triples the previous protein compatibility, but also fundamentally extends the processes observable by DSF, including interdomain allostery in O-GlcNAc Transferase (OGT). paDSF enables routine measurement of protein stability, dynamics, and ligand binding.

One-Sentence SummaryNext generation protein-adaptive DSF (paDSF) enables rapid and general measurements of protein stability and dynamics.
]]></description>
<dc:creator>Wu, T. S.</dc:creator>
<dc:creator>Yu, J. C.</dc:creator>
<dc:creator>Suresh, A.</dc:creator>
<dc:creator>Gale-Day, Z. J.</dc:creator>
<dc:creator>Alteen, M. G.</dc:creator>
<dc:creator>Woo, A. S.</dc:creator>
<dc:creator>Millbern, Z.</dc:creator>
<dc:creator>Johnson, O. T.</dc:creator>
<dc:creator>Carroll, E. C.</dc:creator>
<dc:creator>Partch, C. L.</dc:creator>
<dc:creator>Fourches, D.</dc:creator>
<dc:creator>Vilnueza, N. R.</dc:creator>
<dc:creator>Vocadlo, D. J.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:date>2023-01-24</dc:date>
<dc:identifier>doi:10.1101/2023.01.23.525251</dc:identifier>
<dc:title><![CDATA[Conformationally Adaptive Dyes Enable Protein-Adaptive Differential Scanning Fluorometry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.24.525392v1?rss=1">
<title>
<![CDATA[
Cholecystokinin-A Signaling Regulates Automaticity of Pacemaker Cardiomyocytes and Shortens Sinus Node Recovery Time 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.24.525392v1?rss=1</link>
<description><![CDATA[
AimsThe behavior of pacemaker cardiomyocytes (PCs) in the sinoatrial node (SAN) is modulated by neurohormonal and paracrine factors, many of which signal through G-protein coupled receptors (GPCRs). The aims of the present study are to catalog GPCRs that are differentially expressed in the mammalian SAN and to define the acute physiological consequences of activating the cholecystokinin-A signaling system in isolated PCs.

Methods and ResultsUsing bulk and single cell RNA sequencing datasets, we identify a set of GPCRs that are differentially expressed between SAN and right atrial tissue, including several whose roles in PCs and in the SAN have not been thoroughly characterized. Focusing on one such GPCR, Cholecystokinin-A receptor (CCKAR), we demonstrate expression of Cckar mRNA specifically in mouse PCs, and further demonstrate that subsets of SAN fibroblasts and neurons within the cardiac intrinsic nervous system express cholecystokinin, the ligand for CCKAR. Using mouse models, we find that while baseline SAN function is not dramatically affected by loss of CCKAR, the firing rate of individual PCs is slowed by exposure to sulfated cholecystokinin-8 (sCCK-8), the high affinity ligand for CCKAR. The effect of sCCK-8 on firing rate is mediated by reduction in the rate of spontaneous phase 4 depolarization of PCs and is mitigated by activation of beta-adrenergic signaling.

Conclusions(1) PCs express many GPCRs whose specific roles in SAN function have not been characterized, (2) Activation of the the cholecystokinin-A signaling pathway regulates PC automaticity.
]]></description>
<dc:creator>Ruan, H.</dc:creator>
<dc:creator>Mandla, R.</dc:creator>
<dc:creator>Ravi, N.</dc:creator>
<dc:creator>Galang, G.</dc:creator>
<dc:creator>Soe, A. W.</dc:creator>
<dc:creator>Olgin, J. E.</dc:creator>
<dc:creator>Lang, D.</dc:creator>
<dc:creator>Vedantham, V.</dc:creator>
<dc:date>2023-01-24</dc:date>
<dc:identifier>doi:10.1101/2023.01.24.525392</dc:identifier>
<dc:title><![CDATA[Cholecystokinin-A Signaling Regulates Automaticity of Pacemaker Cardiomyocytes and Shortens Sinus Node Recovery Time]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.25.525428v1?rss=1">
<title>
<![CDATA[
Structural variation across 138,134 samples in the TOPMed consortium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.25.525428v1?rss=1</link>
<description><![CDATA[
Ever larger Structural Variant (SV) catalogs highlighting the diversity within and between populations help researchers better understand the links between SVs and disease. The identification of SVs from DNA sequence data is non-trivial and requires a balance between comprehensiveness and precision. Here we present a catalog of 355,667 SVs (59.34% novel) across autosomes and the X chromosome (50bp+) from 138,134 individuals in the diverse TOPMed consortium. We describe our methodologies for SV inference resulting in high variant quality and >90% allele concordance compared to long-read de-novo assemblies of well-characterized control samples. We demonstrate utility through significant associations between SVs and important various cardio-metabolic and hemotologic traits. We have identified 690 SV hotspots and deserts and those that potentially impact the regulation of medically relevant genes. This catalog characterizes SVs across multiple populations and will serve as a valuable tool to understand the impact of SV on disease development and progression.
]]></description>
<dc:creator>Jun, G.</dc:creator>
<dc:creator>English, A. C.</dc:creator>
<dc:creator>Metcalf, G. A.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Chaisson, M. J.</dc:creator>
<dc:creator>Pankratz, N.</dc:creator>
<dc:creator>Menon, V. K.</dc:creator>
<dc:creator>Salerno, W. J.</dc:creator>
<dc:creator>Krasheninina, O.</dc:creator>
<dc:creator>Smith, A. V.</dc:creator>
<dc:creator>Lane, J. A.</dc:creator>
<dc:creator>Blackwell, T.</dc:creator>
<dc:creator>Kang, H. M.</dc:creator>
<dc:creator>Salvi, S.</dc:creator>
<dc:creator>Meng, Q.</dc:creator>
<dc:creator>Shen, H.</dc:creator>
<dc:creator>Pasham, D.</dc:creator>
<dc:creator>Bhamidipati, S.</dc:creator>
<dc:creator>Kottapalli, K.</dc:creator>
<dc:creator>Arnett, D. K.</dc:creator>
<dc:creator>Ashley-Koch, A.</dc:creator>
<dc:creator>Auer, P. L.</dc:creator>
<dc:creator>Beutel, K. M.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Bowden, D. W.</dc:creator>
<dc:creator>Brody, J. A.</dc:creator>
<dc:creator>Cade, B. E.</dc:creator>
<dc:creator>Chen, Y.-D. I.</dc:creator>
<dc:creator>Cho, M. H.</dc:creator>
<dc:creator>Curran, J. E.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Freedman, B. I.</dc:creator>
<dc:creator>Fingerlin, T.</dc:creator>
<dc:creator>Gelb, B. D.</dc:creator>
<dc:creator>Hou, L.</dc:creator>
<dc:creator>Hung, Y.-J.</dc:creator>
<dc:creator>Kane, J. P.</dc:creator>
<dc:creator>Kaplan, R.</dc:creator>
<dc:creator>Kim, W.</dc:creator>
<dc:creator>Loos, R. J. F.</dc:creator>
<dc:creator>Marcus,, G. M.</dc:creator>
<dc:creator>Mathias, R. A.</dc:creator>
<dc:creator>McGarv</dc:creator>
<dc:date>2023-01-26</dc:date>
<dc:identifier>doi:10.1101/2023.01.25.525428</dc:identifier>
<dc:title><![CDATA[Structural variation across 138,134 samples in the TOPMed consortium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.25.525576v1?rss=1">
<title>
<![CDATA[
Patterns of within-host spread of Chlamydia trachomatis between vagina, endocervix and rectum revealed by comparative genomic analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.25.525576v1?rss=1</link>
<description><![CDATA[
Chlamydia trachomatis, a gram-negative obligate intracellular bacterium, commonly causes sexually transmitted infections (STIs). Little is known about C. trachomatis transmission within the host, which is important for understanding disease epidemiology and progression. We used RNA-bait enrichment and whole-genome sequencing to compare rectal, vaginal and endocervical samples collected at the same time from 26 study participants who attended Fijian Ministry of Health and Medical Services clinics and tested positive for C. trachomatis at each anatomic site. The 78 C. trachomatis genomes from participants were from two major clades of the C. trachomatis phylogeny (the "prevalent urogenital and anorecta"l clade and "non-prevalent urogenital and anorectal" clade). For 21 participants, genome sequences were almost identical in each anatomic site. For the other five participants, two distinct C. trachomatis strains were present in different sites; in two cases, the vaginal sample was a mixture of strains. The absence of large numbers of fixed SNPs between C. trachomatis strains within many of the participants could indicate recent acquisition of infection prior to the clinic visit without sufficient time to accumulate significant variation in the different body sites. This model suggests that many C. trachomatis infections may be resolved relatively quickly in the Fijian population, possibly reflecting common prescription or over-the-counter antibiotics usage.

ImportanceChlamydia trachomatis is a bacterial pathogen that causes millions of sexually transmitted infections (STIs) annually across the globe. Because C. trachomatis lives inside human cells, it has historically been hard to study. We know little about how the bacterium spreads between body sites. Here, samples from 26 study participants who had simultaneous infections in their vagina, rectum and endocervix were genetically analyzed using an improved method to extract C. trachomatis DNA directly from clinical samples for genome sequencing. By analyzing patterns of mutations in the genomes, we found that 21 participants shared very similar C. trachomatis strains in all three anatomic sites, suggesting recent infection and spread. For five participants two C. trachomatis strains were evident, indicating multiple infections. This study is significant in that improved enrichment methods for genome sequencing provides robust data to genetically trace patterns of C. trachomatis infection and transmission within an individual for epidemiologic and pathogenesis interrogations.
]]></description>
<dc:creator>Joseph, S. J.</dc:creator>
<dc:creator>Bommona, S.</dc:creator>
<dc:creator>Ziklo, N.</dc:creator>
<dc:creator>Kama, M.</dc:creator>
<dc:creator>Dean, D.</dc:creator>
<dc:creator>Read, T. D.</dc:creator>
<dc:date>2023-01-26</dc:date>
<dc:identifier>doi:10.1101/2023.01.25.525576</dc:identifier>
<dc:title><![CDATA[Patterns of within-host spread of Chlamydia trachomatis between vagina, endocervix and rectum revealed by comparative genomic analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.27.525867v1?rss=1">
<title>
<![CDATA[
Mass cytometry identifies dysregulation of circulating and cutaneous immune cells in patients with hidradenitis suppurativa 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.27.525867v1?rss=1</link>
<description><![CDATA[
2.BackgroundHidradenitis suppurativa (HS) is a multifactorial, inflammatory skin disease. Increased systemic inflammatory comorbidities and serum cytokines highlight systemic inflammation as a feature of HS. However, the specific immune cell subsets contributing to systemic and cutaneous inflammation have not been resolved.

ObjectiveIdentify features of peripheral and cutaneous immune dysregulation.

MethodsHere, we generated whole-blood immunomes by mass cytometry. We performed a meta-analysis of RNA-seq data, immunohistochemistry, and imaging mass cytometry to characterize the immunological landscape of skin lesions and perilesions from patients with HS.

ResultsBlood from patients with HS exhibited lower frequencies of natural killer cells, dendritic cells, and classical (CD14+CD16-) and nonclassical (CD14-CD16+) monocytes, as well as higher frequencies of Th17 cells and intermediate (CD14+CD16+) monocytes than blood from healthy controls. Classical and intermediate monocytes from patients with HS had increased expression of skin-homing chemokine receptors. Furthermore, we identified a CD38+ intermediate monocyte subpopulation that was more abundant in the immunome of blood from patients with HS. Meta-analysis of RNA-seq data found higher CD38 expression in lesional HS skin than in perilesional skin, and markers of classical monocyte infiltration. Imaging mass cytometry showed that CD38+ classical monocytes and CD38+ monocyte-derived macrophages were more abundant in lesional HS skin.

ConclusionOverall, we report targeting CD38 may be worth pursuing in clinical trials.

3. Key MessagesO_LIMonocyte subsets express markers of activation in circulation and HS lesions
C_LIO_LITargeting CD38 may be a viable strategy for treating systemic and cutaneous inflammation in patients with HS
C_LI

4. Capsule SummaryDysregulated immune cells in patients with HS express CD38 and may be targeting by anti-CD38 immunotherapy.
]]></description>
<dc:creator>Dimitrion, P.</dc:creator>
<dc:creator>Hamzavi, I.</dc:creator>
<dc:creator>Yin, C.</dc:creator>
<dc:creator>Loveless, I.</dc:creator>
<dc:creator>Toor, J.</dc:creator>
<dc:creator>Subedi, K.</dc:creator>
<dc:creator>Khalasawi, N.</dc:creator>
<dc:creator>Miller, A.</dc:creator>
<dc:creator>Huggins, R.</dc:creator>
<dc:creator>Adrianto, I.</dc:creator>
<dc:creator>Veenstra, J.</dc:creator>
<dc:creator>Vellaichamy, G.</dc:creator>
<dc:creator>Hans, A.</dc:creator>
<dc:creator>Daveluy, S.</dc:creator>
<dc:creator>Athar, M.</dc:creator>
<dc:creator>Liao, W.</dc:creator>
<dc:creator>Lim, H.</dc:creator>
<dc:creator>Ozog, D.</dc:creator>
<dc:creator>Zhou, L.</dc:creator>
<dc:creator>Mi, Q.-S.</dc:creator>
<dc:date>2023-01-28</dc:date>
<dc:identifier>doi:10.1101/2023.01.27.525867</dc:identifier>
<dc:title><![CDATA[Mass cytometry identifies dysregulation of circulating and cutaneous immune cells in patients with hidradenitis suppurativa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.27.525930v1?rss=1">
<title>
<![CDATA[
Structure and dynamics of the essential endogenous mycobacterial polyketide synthasePks13 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.27.525930v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis is currently the leading cause of death by any bacterial infection1. The mycolic acid layer of the cell wall is essential for viability and virulence, and the enzymes responsible for its synthesis are therefore front line targets for antimycobacterial drug development2,3. Polyketide synthase 13 (Pks13) is a module comprised of a closely symmetric parallel dimer of chains, each encoding several enzymatic and transport functions, that carries out the condensation of two different very long chain fatty acids to produce mycolic acids that are essential components of the mycobacterial cell wall. Consequently individual enzymatic domains of Pks13 are targets for antimycobacterial drug development4. To understand this machinery, we sought to determine the structure and domain trajectories of the dimeric multi-enzyme Pks13, a 2x198,426 Dalton complex, from protein purified endogenously from mycobacteria under normal growth conditions, to capture it with normal substrates bound trapped  in action.

Structures of the multi-domain assembly revealed by cryogenic electron microscopy (cryoEM) define the ketosynthase (KS), linker, and acyltransferase (AT) domains, each at atomic resolution (1.8[A]), with bound substrates defined at 2.4[A] and 2.9[A] resolution. Image classification reveals two distinct structures with alternate locations of the N-terminal acyl carrier protein (termed ACP1a, ACP1b) seen at 3.6[A] and 4.6[A] resolution respectively. These two structures suggest plausible intermediate states, related by a ~60[A] movement of ACP1, on the pathway for substrate delivery from the fatty acyl-ACP ligase (FadD32) to the ketosynthase domain. The linking sequence between ACP1 and the KS includes an 11 amino acid sequence with 6 negatively charged side chains that lies in different positively charged grooves on the KS in ACP1a versus ACP1b structures. This charge complementarity between the extended chain and the grooves suggests some stabilization of these two distinct orientations. Other domains are visible at lower resolution and indicate flexibility relative to the KS-AT core. The chemical structures of three bound endogenous long chain fatty acid substrates with their proximal regions defined in the structures were determined by electrospray ionization mass spectrometry.

The domain proximities were probed by chemical cross-linking and identified by mass spectrometry. These were incorporated into integrative structure modeling to define multiple domain configurations that transport the very long fatty acid chains throughout the multistep Pks13 mediated synthetic pathway.
]]></description>
<dc:creator>Kim, S. K.</dc:creator>
<dc:creator>Dickinson, M. S.</dc:creator>
<dc:creator>Finer-Moore, J. S.</dc:creator>
<dc:creator>Guan, Z.</dc:creator>
<dc:creator>Kaake, R. M.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Pulido, E. H.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Rosenberg, O. S.</dc:creator>
<dc:creator>Stroud, R. M.</dc:creator>
<dc:date>2023-01-28</dc:date>
<dc:identifier>doi:10.1101/2023.01.27.525930</dc:identifier>
<dc:title><![CDATA[Structure and dynamics of the essential endogenous mycobacterial polyketide synthasePks13]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.29.526112v1?rss=1">
<title>
<![CDATA[
A Systematic Approach to the Discovery of Protein-Protein Inter-action Stabilizers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.29.526112v1?rss=1</link>
<description><![CDATA[
Protein-protein interactions (PPIs) are responsible for the proper function of biological processes and, when dysregulated, commonly lead to disease. PPI stabilization has only recently been systematically explored for drug discovery despite being a powerful approach to selectively target intrinsically disordered proteins and hub proteins, like 14-3-3, with multiple interaction partners. Disulfide tethering is a site-directed fragment-based drug discovery (FBDD) methodology for screening small molecules in a quantitative, high-throughput manner. We explore the scope of the disulfide tethering technology for the discovery of selective fragments as starting points for the development of potent small molecule PPI stabilizers and molecular glues using the hub protein 14-3-3{sigma}. The complexes with 5 biologically and structurally diverse phospho-peptides, derived from the 14-3-3 client proteins ER, FOXO1, C-RAF, USP8, and SOS1, were screened for hit identification. Stabilizing fragments could be found for 4/5 client complexes with a diversified hit-rate and stabilizing efficacy for the different 14-3-3/client phospho-peptides. Extensive structural elucidation revealed the ability and adaptivity of the peptide to make productive interactions with the tethered fragments as key criterion for cooperative complex formation. We validated eight fragment stabilizers, six of which showed selectivity for one phospho-peptide client, and structurally characterized two nonselective hits and four fragments that selectively stabilized C-RAF or FOXO1. The most efficacious of these fragments increased 14-3-3{sigma}/C-RAF phospho-peptide affinity by 430-fold. Disulfide tethering to the wildtype C38 in 14-3-3{sigma} provided diverse structures for future optimization of 14-3-3/client stabilizers and highlighted a systematic method to discover molecular glues.
]]></description>
<dc:creator>Kenanova, D. N.</dc:creator>
<dc:creator>Visser, E. J.</dc:creator>
<dc:creator>Virta, J. M.</dc:creator>
<dc:creator>Sijbesma, E.</dc:creator>
<dc:creator>Centorrino, F.</dc:creator>
<dc:creator>Vickery, H. R.</dc:creator>
<dc:creator>Zhong, M.</dc:creator>
<dc:creator>Neitz, J. R.</dc:creator>
<dc:creator>Brunsveld, L.</dc:creator>
<dc:creator>Ottmann, C.</dc:creator>
<dc:creator>Arkin, M. R.</dc:creator>
<dc:date>2023-01-30</dc:date>
<dc:identifier>doi:10.1101/2023.01.29.526112</dc:identifier>
<dc:title><![CDATA[A Systematic Approach to the Discovery of Protein-Protein Inter-action Stabilizers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.30.526384v1?rss=1">
<title>
<![CDATA[
Mass Spectrometry Imaging of Hair Identifies Daily Maraviroc Adherence in HPTN 069/ACTG A5305 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.30.526384v1?rss=1</link>
<description><![CDATA[
Objective measures of adherence for antiretrovirals used as pre-exposure prophylaxis (PrEP) are critical for improving preventative efficacy in both clinical trials and real-world application. Current objective adherence measures either reflect only recent behavior (eg days for plasma or urine) or cumulative behavior (eg months for dried blood spots). We measured the accumulation of the antiretroviral drug maraviroc (MVC) in hair strands by infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry imaging (MSI) to evaluate adherence behavior longitudinally at high temporal resolution. An MSI threshold for classifying daily adherence was established using clinical samples from healthy volunteers following directly observed dosing of 1 to 7 doses MVC/week. We then used the benchmarked MSI assay to classify adherence to MVC-based PrEP regimens in hair samples collected throughout the 49-week HPTN069/ACTGA5305 study. We found that only ~32% of investigated hair samples collected during the studys active dosing period showed consistent daily PrEP adherence throughout a retrospective period of 30 days, and also found that profiles of daily individual adherence from MSI hair analysis could identify when patients were and were not taking study drug. The assessment of adherence from MSI hair strand analysis was 62% lower than adherence classified using paired plasma samples, the latter of which may be influenced by white-coat adherence. These findings demonstrate the ability of MSI hair analysis to examine daily variability of adherence behavior over a longer-term measurement and offer the potential for longitudinal comparison with risk behavior to target patient-specific adherence interventions and improve outcomes.
]]></description>
<dc:creator>Rosen, E. P.</dc:creator>
<dc:creator>White, N. R.</dc:creator>
<dc:creator>Gilliland, W. M.</dc:creator>
<dc:creator>Gerona, R.</dc:creator>
<dc:creator>Gandhi, M.</dc:creator>
<dc:creator>Amico, K. R.</dc:creator>
<dc:creator>Mayer, K. H.</dc:creator>
<dc:creator>Gulick, R. M.</dc:creator>
<dc:creator>Kashuba, A. D. M.</dc:creator>
<dc:date>2023-02-02</dc:date>
<dc:identifier>doi:10.1101/2023.01.30.526384</dc:identifier>
<dc:title><![CDATA[Mass Spectrometry Imaging of Hair Identifies Daily Maraviroc Adherence in HPTN 069/ACTG A5305]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.31.526482v1?rss=1">
<title>
<![CDATA[
A thin-film optogenetic visual prosthesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.31.526482v1?rss=1</link>
<description><![CDATA[
Retinitis pigmentosa and macular degeneration lead to photoreceptor death and loss of visual perception. Despite recent progress, restorative technologies for photoreceptor degeneration remain largely unavailable. Here, we describe a novel optogenetic visual prosthesis (FlexLED) based on a combination of a thin-film retinal display and optogenetic activation of retinal ganglion cells (RGCs). The FlexLED implant is a 30 {micro}m thin, flexible, wireless {micro}LED display with 8,192 pixels, each with an emission area of 66 {micro}m2. The display is affixed to the retinal surface, and the electronics package is mounted under the conjunctiva in the form factor of a conventional glaucoma drainage implant. In a rabbit model of photoreceptor degeneration, optical stimulation of the retina using the FlexLED elicits activity in visual cortex. This technology is readily scalable to hundreds of thousands of pixels, providing a route towards an implantable optogenetic visual prosthesis capable of generating vision by stimulating RGCs at near-cellular resolution.
]]></description>
<dc:creator>Knudsen, E. B.</dc:creator>
<dc:creator>Zappitelli, K.</dc:creator>
<dc:creator>Brown, J.</dc:creator>
<dc:creator>Reeder, J.</dc:creator>
<dc:creator>Smith, K. S.</dc:creator>
<dc:creator>Rostov, M.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Rochford, A.</dc:creator>
<dc:creator>Slager, N.</dc:creator>
<dc:creator>Miura, S. K.</dc:creator>
<dc:creator>Rodgers, K.</dc:creator>
<dc:creator>Reed, A.</dc:creator>
<dc:creator>Lewis Israeli, Y. R.</dc:creator>
<dc:creator>Shiraga, S.</dc:creator>
<dc:creator>Seo, K. J.</dc:creator>
<dc:creator>Wolin, C.</dc:creator>
<dc:creator>Dawson, P.</dc:creator>
<dc:creator>Eltaeb, M.</dc:creator>
<dc:creator>Dasgupta, A.</dc:creator>
<dc:creator>Chong, P.</dc:creator>
<dc:creator>Charles, S.</dc:creator>
<dc:creator>Stewart, J. M.</dc:creator>
<dc:creator>Silva, R. A.</dc:creator>
<dc:creator>Kim, T.</dc:creator>
<dc:creator>Kong, Y.</dc:creator>
<dc:creator>Mardinly, A. R.</dc:creator>
<dc:creator>Hodak, M.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.01.31.526482</dc:identifier>
<dc:title><![CDATA[A thin-film optogenetic visual prosthesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.01.526692v1?rss=1">
<title>
<![CDATA[
EMBER multi-dimensional spectral microscopy enables quantitative determination of disease- and cell-specific amyloid strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.01.526692v1?rss=1</link>
<description><![CDATA[
In neurodegenerative diseases proteins fold into amyloid structures with distinct conformations (strains) that are characteristic of different diseases. However, there is a need to rapidly identify amyloid conformations in situ. Here we use machine learning on the full information available in fluorescent excitation/emission spectra of amyloid binding dyes to identify six distinct different conformational strains in vitro, as well as A{beta} deposits in different transgenic mouse models. Our EMBER (excitation multiplexed bright emission recording) imaging method rapidly identifies conformational differences in A{beta} and tau deposits from Down syndrome, sporadic and familial Alzheimers disease human brain slices. EMBER has in situ identified distinct conformational strains of tau inclusions in astrocytes, oligodendrocytes, and neurons from Picks disease. In future studies, EMBER should enable high-throughput measurements of the fidelity of strain transmission in cellular and animal neurodegenerative diseases models, time course of amyloid strain propagation, and identification of pathogenic versus benign strains.

SignificanceIn neurodegenerative diseases proteins fold into amyloid structures with distinct conformations (strains) that are characteristic of different diseases. There is a need to rapidly identify these amyloid conformations in situ. Here we use machine learning on the full information available in fluorescent excitation/emission spectra of amyloid binding dyes to identify six distinct different conformational strains in vitro, as well as A{beta} deposits in different transgenic mouse models. Our imaging method rapidly identifies conformational differences in A{beta} and tau deposits from Down syndrome, sporadic and familial Alzheimers disease human brain slices. We also identified distinct conformational strains of tau inclusions in astrocytes, oligodendrocytes, and neurons from Picks disease. These findings will facilitate the identification of pathogenic protein aggregates to guide research and treatment of protein misfolding diseases.
]]></description>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Yuan, P.</dc:creator>
<dc:creator>Wu, y.</dc:creator>
<dc:creator>Shi, M.</dc:creator>
<dc:creator>caro, C.</dc:creator>
<dc:creator>tengeiji, a.</dc:creator>
<dc:creator>yamanoi, s.</dc:creator>
<dc:creator>Inoue, M.</dc:creator>
<dc:creator>DeGrado, W.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.01.526692</dc:identifier>
<dc:title><![CDATA[EMBER multi-dimensional spectral microscopy enables quantitative determination of disease- and cell-specific amyloid strains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.01.526706v1?rss=1">
<title>
<![CDATA[
Optimizing anesthesia and delivery approaches for dosing into lungs of mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.01.526706v1?rss=1</link>
<description><![CDATA[
Microbes, toxins, therapeutics and cells are often instilled into lungs of mice to model diseases and test experimental interventions. Consistent pulmonary delivery is critical for experimental power and reproducibility, but we observed variation in outcomes between handlers using different anesthetic approaches for intranasal dosing into mice. We therefore used a radiotracer to quantify lung delivery after intranasal dosing under inhalational (isoflurane) versus injectable (ketamine/xylazine) anesthesia in C57BL/6 mice. We found that ketamine/xylazine anesthesia resulted in delivery of a greater proportion (52{+/-}9%) of an intranasal dose to lungs relative to isoflurane anesthesia (30{+/-}15%). This difference in pulmonary dose delivery altered key outcomes in models of viral and bacterial pneumonia, with mice anesthetized with ketamine/xylazine for intranasal infection with influenza A virus or Pseudomonas aeruginosa developing more robust lung inflammation responses relative to control animals randomized to isoflurane anesthesia. Pulmonary dosing efficiency through oropharyngeal aspiration was not affected by anesthetic method and resulted in delivery of 63{+/-}8% of dose to lungs, and a non-surgical intratracheal dosing approach further increased lung delivery to 92{+/-}6% of dose. Use of either of these more precise dosing methods yielded greater experimental power in the bacterial pneumonia model relative to intranasal infection. Both anesthetic approach and dosing route can impact pulmonary dosing efficiency. These factors affect experimental power and so should be considered when planning and reporting studies involving delivery of fluids to lungs of mice.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=158 SRC="FIGDIR/small/526706v2_ufig1.gif" ALT="Figure 1">
View larger version (51K):
org.highwire.dtl.DTLVardef@10e90forg.highwire.dtl.DTLVardef@34f8a5org.highwire.dtl.DTLVardef@1b02406org.highwire.dtl.DTLVardef@10d3f33_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>Qiu, L.</dc:creator>
<dc:creator>Magnen, M.</dc:creator>
<dc:creator>Conrad, C.</dc:creator>
<dc:creator>Moussavi-Harami, S. F.</dc:creator>
<dc:creator>Looney, M. R.</dc:creator>
<dc:creator>Cleary, S. J.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.01.526706</dc:identifier>
<dc:title><![CDATA[Optimizing anesthesia and delivery approaches for dosing into lungs of mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.03.526065v1?rss=1">
<title>
<![CDATA[
Clinically relevant concurrent BRAF and MEK inhibition alters differentiation states and sensitizes BRAF V600E-mutated high-grade gliomas to immune checkpoint blockade 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.03.526065v1?rss=1</link>
<description><![CDATA[
Resistance to BRAF plus MEK inhibition (BRAFi+MEKi) in BRAFV600E-mutant gliomas drives rebound, progression, and high mortality, yet it remains poorly understood. This study addresses the urgent need to develop treatments for BRAFi+MEKi-resistant glioma in novel mouse models and patient-derived materials. BRAFi+MEKi reveals glioma plasticity by heightening cell state transitions along glial differentiation trajectories, giving rise to astrocyte- and immunomodulatory oligodendrocyte (OL)-like states. PD-L1 upregulation in OL-like cells links cell state transitions to tumor evasion, possibly orchestrated by Galectin-3. BRAFi+MEKi induces interferon response signatures, tumor infiltration, and suppression of T cells. Combining BRAFi+MEKi with immune checkpoint inhibition enhances survival in a T cell-dependent manner, reinvigorates T cells, and outperforms individual or sequential therapies in mice. Elevated PD-L1 expression in BRAF-mutant versus BRAF-wildtype glioblastoma supports the rationale for PD-1 inhibition in patients. These findings underscore the potential of targeting glioma plasticity and highlight combination strategies to overcome therapy resistance in BRAFV600E-mutant HGG.

In briefXing et al. show that combined BRAF and MEK inhibitor (BRAFi+MEKi) treatment induces cell state transitions in BRAFV600E-mutant high-grade glioma cells linked with programmed death-ligand (PD-L1) upregulation and T cell suppression, potentially orchestrated through the secretion of galectin-3. These tumor-intrinsic adaptations may be overcome by concurrent immune checkpoint inhibition (ICI), as demonstrated in murine studies, offering novel therapeutic opportunities.

HighlightsO_LIBRAFV600E-mutant HGG exhibits cell plasticity induced by BRAFi+MEKi, which links cell state transitions towards glial differentiation with immune evasion
C_LIO_LIBRAFi+MEKi enhances anti-tumor immunity and simultaneously suppresses T cells via PD-L1 upregulation
C_LIO_LIBRAF-mutant glioblastoma has elevated PD-L1 expression compared to BRAF-wildtype counterparts, providing a criterion for PD-1 inhibition therapy
C_LIO_LIConcurrent BRAFi+MEKi and immune checkpoint inhibition enhance T cell-mediated anti-tumor activity and boost survival more effectively than sequential treatment in mice, guiding clinical translation
C_LI
]]></description>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Grossauer, S.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Xing, Y. L.</dc:creator>
<dc:creator>Koeck, K.</dc:creator>
<dc:creator>Garcia, C. A.</dc:creator>
<dc:creator>Nasajpour, E.</dc:creator>
<dc:creator>Wilson, C.</dc:creator>
<dc:creator>Lancero, H.</dc:creator>
<dc:creator>Harter, P. N.</dc:creator>
<dc:creator>Filipski, K.</dc:creator>
<dc:creator>Daynac, M.</dc:creator>
<dc:creator>Meyer, L.</dc:creator>
<dc:creator>Guinle, M. I. B.</dc:creator>
<dc:creator>Monje, M.</dc:creator>
<dc:creator>Vogel, H.</dc:creator>
<dc:creator>Lim, M.</dc:creator>
<dc:creator>Prolo, L. M.</dc:creator>
<dc:creator>Grant, G. A.</dc:creator>
<dc:creator>Petritsch, C. K.</dc:creator>
<dc:date>2023-02-04</dc:date>
<dc:identifier>doi:10.1101/2023.02.03.526065</dc:identifier>
<dc:title><![CDATA[Clinically relevant concurrent BRAF and MEK inhibition alters differentiation states and sensitizes BRAF V600E-mutated high-grade gliomas to immune checkpoint blockade]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.03.526887v1?rss=1">
<title>
<![CDATA[
The Clustered Gamma Protocadherin Pcdhγc4 Isoform Regulates Cortical Interneuron Programmed Cell Death in the Mouse Cortex. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.03.526887v1?rss=1</link>
<description><![CDATA[
Cortical function critically depends on inhibitory/excitatory balance. Cortical inhibitory interneurons (cINs) are born in the ventral forebrain and migrate into cortex, where their numbers are adjusted by programmed cell death. Previously, we showed that loss of clustered gamma protocadherins (Pcdh{gamma}), but not of genes in the alpha or beta clusters, increased dramatically cIN BAX-dependent cell death in mice. Here we show that the sole deletion of the Pcdh{gamma}c4 isoform, but not of the other 21 isoforms in the Pcdh{gamma} gene cluster, increased cIN cell death in mice during the normal period of programmed cell death. Viral expression of the Pcdh{gamma}c4 isoform rescued transplanted cINs lacking Pcdh{gamma} from cell death. We conclude that Pcdh{gamma}, specifically Pcdh{gamma}c4, plays a critical role in regulating the survival of cINs during their normal period of cell death. This demonstrates a novel specificity in the role of Pcdh{gamma} isoforms in cortical development.
]]></description>
<dc:creator>Mancia Leon, W. R.</dc:creator>
<dc:creator>Steffen, D. M.</dc:creator>
<dc:creator>Dale-Huang, F.</dc:creator>
<dc:creator>Rakela, B.</dc:creator>
<dc:creator>Breevoort, A.</dc:creator>
<dc:creator>Romero-Rodriguez, R.</dc:creator>
<dc:creator>Hasenstaub, A. R.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:creator>Weiner, J. A.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:date>2023-02-04</dc:date>
<dc:identifier>doi:10.1101/2023.02.03.526887</dc:identifier>
<dc:title><![CDATA[The Clustered Gamma Protocadherin Pcdhγc4 Isoform Regulates Cortical Interneuron Programmed Cell Death in the Mouse Cortex.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.03.527061v1?rss=1">
<title>
<![CDATA[
Neanderthal Introgression Shaped Human Circadian Traits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.03.527061v1?rss=1</link>
<description><![CDATA[
IntroductionWhen the ancestors of modern Eurasians migrated out of Africa and interbred with Eurasian archaic hominins, namely Neanderthals and Denisovans, DNA of archaic ancestry integrated into the genomes of anatomically modern humans. This process potentially accelerated adaptation to Eurasian environmental factors, including reduced ultra-violet radiation and increased variation in seasonal dynamics. However, whether these groups differed substantially in circadian biology, and whether archaic introgression adaptively contributed to human chronotypes remains unknown.

ResultsHere we traced the evolution of chronotype based on genomes from archaic hominins and present-day humans. First, we inferred differences in circadian gene sequences, splicing, and regulation between archaic hominins and modern humans. We identified 28 circadian genes containing variants with potential to alter splicing in archaics (e.g., CLOCK, PER2, RORB, RORC), and 16 circadian genes likely divergently regulated between present-day humans and archaic hominins, including RORA. These differences suggest the potential for introgression to modify circadian gene expression. Testing this hypothesis, we found that introgressed variants are enriched among eQTLs for circadian genes. Supporting the functional relevance of these regulatory effects, we found that many introgressed alleles have associations with chronotype. Strikingly, the strongest introgressed effects on chronotype increase morningness, consistent with adaptations to high latitude in other species. Finally, we identified several circadian loci with evidence of adaptive introgression or latitudinal clines in allele frequency.

ConclusionsThese findings identify differences in circadian gene regulation between modern humans and archaic hominins and support the contribution of introgression via coordinated effects on variation in human chronotype.

SIGNIFICANCE STATEMENTInterbreeding between humans and Neanderthals created the potential for adaptive introgression as humans moved into environments that had been populated by Neanderthals for hundreds of thousands of years. Here we discover lineage-specific genetic differences in circadian genes and their regulatory elements between humans and Neanderthals. We show that introgressed alleles are enriched for effects on circadian gene regulation, consistently increase propensity for morningness in Europeans, and show evidence of adaptive introgression or associations between latitude and frequency. These results expand our understanding of how the genomes of humans and our closest relatives responded to environments with different light/dark cycles, and demonstrate a coordinated contribution of admixture to human chronotype in a direction that is consistent with adaptation to higher latitudes.
]]></description>
<dc:creator>Velazquez-Arcelay, K.</dc:creator>
<dc:creator>Colbran, L. L.</dc:creator>
<dc:creator>McArthur, E.</dc:creator>
<dc:creator>Brand, C. M.</dc:creator>
<dc:creator>Siemann, J. K.</dc:creator>
<dc:creator>McMahon, D. G.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:date>2023-02-04</dc:date>
<dc:identifier>doi:10.1101/2023.02.03.527061</dc:identifier>
<dc:title><![CDATA[Neanderthal Introgression Shaped Human Circadian Traits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.03.527062v1?rss=1">
<title>
<![CDATA[
Lung microenvironments harbor  Mycobacterium tuberculosis phenotypes with distinct treatment responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.03.527062v1?rss=1</link>
<description><![CDATA[
Tuberculosis lung lesions are complex and harbor heterogeneous microenvironments that influence antibiotic effectiveness. Major strides have been made recently in understanding drug pharmacokinetics in pulmonary lesions, but the bacterial phenotypes that arise under these conditions and their contribution to drug tolerance is poorly understood. A pharmacodynamic marker called the RS ratio quantifies ongoing rRNA synthesis based on the abundance of newly-synthesized precursor rRNA relative to mature structural rRNA. Application of the RS ratio in the C3HeB/FeJ mouse model demonstrated that Mycobacterium tuberculosis populations residing in different tissue microenvironments are phenotypically distinct and respond differently to drug treatment with rifampin, isoniazid or bedaquiline. This work provides a foundational basis required to address how anatomic and pathologic microenvironmental niches may contribute to the long treatment duration and drug tolerance during treatment of human tuberculosis.
]]></description>
<dc:creator>Walter, N. D.</dc:creator>
<dc:creator>Ernest, J. P.</dc:creator>
<dc:creator>Dide-Agossou, C.</dc:creator>
<dc:creator>Bauman, A. A.</dc:creator>
<dc:creator>Ramey, M. E.</dc:creator>
<dc:creator>Rossmassler, K.</dc:creator>
<dc:creator>Massoudi, L. M.</dc:creator>
<dc:creator>Pauly, S.</dc:creator>
<dc:creator>Al Mubarak, R.</dc:creator>
<dc:creator>Voskuil, M. I.</dc:creator>
<dc:creator>Kaya, F.</dc:creator>
<dc:creator>Sarathy, J. P.</dc:creator>
<dc:creator>Zimmerman, M. D.</dc:creator>
<dc:creator>Dartois, V.</dc:creator>
<dc:creator>Podell, B. K.</dc:creator>
<dc:creator>Savic, R. M.</dc:creator>
<dc:creator>Robertson, G. T.</dc:creator>
<dc:date>2023-02-04</dc:date>
<dc:identifier>doi:10.1101/2023.02.03.527062</dc:identifier>
<dc:title><![CDATA[Lung microenvironments harbor  Mycobacterium tuberculosis phenotypes with distinct treatment responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.04.527139v1?rss=1">
<title>
<![CDATA[
Long lifetime and selective accumulation of the A-type lamins accounts for the tissue specificity of Hutchinson-Gilford progeria syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.04.527139v1?rss=1</link>
<description><![CDATA[
Mutations to the LMNA gene cause laminopathies including Hutchinson-Gilford progeria syndrome (HGPS) that severely affect the cardiovascular system. The origins of tissue specificity in these diseases are unclear, as the A-type Lamins are abundant and broadly expressed proteins. We show that A-type Lamin protein and transcript levels are uncorrelated across tissues. As protein-transcript discordance can be caused by variations in protein lifetime, we applied quantitative proteomics to profile protein turnover rates in healthy and progeroid tissues. We discover that tissue context and disease mutation each influence A-type Lamin protein lifetime. Lamin A/C has a weeks-long lifetime in the aorta, heart, and fat, where progeroid pathology is apparent, but a days-long lifetime in the liver and gastrointestinal tract, which are spared from disease. The A-type Lamins are insoluble and densely bundled in cardiovascular tissues, which may present an energetic barrier to degradation and promote long protein lifetime. Progerin is even more long-lived than Lamin A/C in the cardiovascular system and accumulates there over time. Progerin accumulation interferes broadly with protein homeostasis, as hundreds of abundant proteins turn over more slowly in progeroid tissues. These findings indicate that potential gene therapy interventions for HGPS will have significant latency and limited potency in disrupting the long-lived Progerin protein. Finally, we reveal that human disease alleles are significantly over-represented in the long-lived proteome, indicating that long protein lifetime may influence disease pathology and present a significant barrier to gene therapies for numerous human diseases.

Significance statementMany human diseases are caused by mutations to broadly expressed proteins, yet disease mysteriously manifests only in specific tissues. An example of this is Hutchinson-Gilford progeria syndrome (HGPS), which is caused by a mutation to the Lamin A/C protein. We show that this mutation slows the turnover of Lamin A/C proteins in disease-afflicted tissues, causing the mutant "Progerin" protein to accumulate over time and interfere with the normal turnover of hundreds of other proteins. Because Progerin is a long-lived protein, effective therapies for this disease will need to attack the protein and not just the gene that encodes it.
]]></description>
<dc:creator>Hasper, J.</dc:creator>
<dc:creator>Welle, K.</dc:creator>
<dc:creator>Swovick, K.</dc:creator>
<dc:creator>Hryhorenko, J.</dc:creator>
<dc:creator>Ghaemmaghami, S.</dc:creator>
<dc:creator>Buchwalter, A.</dc:creator>
<dc:date>2023-02-04</dc:date>
<dc:identifier>doi:10.1101/2023.02.04.527139</dc:identifier>
<dc:title><![CDATA[Long lifetime and selective accumulation of the A-type lamins accounts for the tissue specificity of Hutchinson-Gilford progeria syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.07.527432v1?rss=1">
<title>
<![CDATA[
Paclitaxel- and vincristine-induced neurotoxicity and drug transport in sensory neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.07.527432v1?rss=1</link>
<description><![CDATA[
Chemotherapy-induced peripheral neuropathy (CIPN) constitutes a significant health problem due to the increasing prevalence and the lack of therapies for treatment and prevention. Patients with CIPN primarily present with sensory symptoms, such as sensory disturbances that may progress to neuropathic pain in hands and feet. While pivotal for routine cancer treatment, paclitaxel and vincristine frequently cause CIPN and impact the quality of life among cancer patients and survivors. We utilized a model of human sensory neurons derived from induced pluripotent stem cells (iPSC-SNs) to provide mechanistic understanding of CIPN caused by paclitaxel and vincristine. The morphological phenotype of iPSC-SNs following paclitaxel exposure was characterized by retraction and thickening of axons while vincristine caused fragmentation and abolishment of axons. Both agents increased the mRNA expression of the pain receptor, transient receptor potential vanilloid (TRPV1), and highly induced neuronal damage, as measured by activating transcription factor 3 (ATF3) mRNA. iPSC-SNs express the efflux transporters, P-glycoprotein (P-gp, encoded by ABCB1) and multidrug resistance-associated protein 1 (MPR1, encoded by ABCC1). Inhibition of P-gp and MRP1 in iPSC-SNs exacerbated neurotoxicity of paclitaxel and vincristine respectively. We further show that pre-treatment with the P-gp inducer rifampicin alleviated chemotherapy-induced structural and transcriptional alterations in iPSC-SNs. iPSC-SNs are a valuable and robust model to study the role of efflux transporters and other mechanistic targets in CIPN. Efflux transporters play a critical role in CIPN pathogenesis as they regulate the disposition of chemotherapy to the peripheral nervous system.
]]></description>
<dc:creator>Mortensen, C.</dc:creator>
<dc:creator>Chua, K. C.</dc:creator>
<dc:creator>Hammer, H. S.</dc:creator>
<dc:creator>Nielsen, F.</dc:creator>
<dc:creator>Poetz, O.</dc:creator>
<dc:creator>Svenningsen, A. F.</dc:creator>
<dc:creator>Kroetz, D. L.</dc:creator>
<dc:creator>Stage, T. B.</dc:creator>
<dc:date>2023-02-07</dc:date>
<dc:identifier>doi:10.1101/2023.02.07.527432</dc:identifier>
<dc:title><![CDATA[Paclitaxel- and vincristine-induced neurotoxicity and drug transport in sensory neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.07.527484v1?rss=1">
<title>
<![CDATA[
IVT generation of guideRNAs for Cas9-enrichment Nanopore Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.07.527484v1?rss=1</link>
<description><![CDATA[
Generating high-coverage sequencing coverage at select genomic loci has extensive applications in both research science and genetic medicine. Long-read sequencing technologies (e.g. nanopore sequencing) have expanded our ability to generate sequencing data in regions (e.g. repetitive elements) that are difficult to interrogate with short-read sequencing methods. In work presented here, we expand on our previous work using CRISPR/Cas9 for targeted nanopore sequencing by using in vitro transcribed guideRNAs, with 1100 guideRNAs in a single experiment. This approach decreases the cost per guideRNA, increases the number of guideRNAs that can be multiplexed in a single experiment, and provides a way to rapidly screen numerous guideRNAs for cutting efficiency. We apply this strategy in multiple patient-derived pancreatic cancer cell lines, demonstrating its ability to unveil structural variation in "deletion hotspots" around the tumor suppressor genes p16 (CDKN2A), and SMAD4.
]]></description>
<dc:creator>Gilpatrick, T.</dc:creator>
<dc:creator>Wang, J. Z.</dc:creator>
<dc:creator>Weiss, D.</dc:creator>
<dc:creator>Norris, A. L.</dc:creator>
<dc:creator>Eshleman, J. R.</dc:creator>
<dc:creator>Timp, W.</dc:creator>
<dc:date>2023-02-07</dc:date>
<dc:identifier>doi:10.1101/2023.02.07.527484</dc:identifier>
<dc:title><![CDATA[IVT generation of guideRNAs for Cas9-enrichment Nanopore Sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.31.525914v1?rss=1">
<title>
<![CDATA[
Rapid assembly of SARS-CoV-2 genomes reveals attenuation of the Omicron BA.1 variant through NSP6 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.31.525914v1?rss=1</link>
<description><![CDATA[
Although the SARS-CoV-2 Omicron variant (BA.1) spread rapidly across the world and effectively evaded immune responses, its viral fitness in cell and animal models was reduced. The precise nature of this attenuation remains unknown as generating replication-competent viral genomes is challenging because of the length of the viral genome (30kb). Here, we designed a plasmid-based viral genome assembly and rescue strategy (pGLUE) that constructs complete infectious viruses or noninfectious subgenomic replicons in a single ligation reaction with >80% efficiency. Fully sequenced replicons and infectious viral stocks can be generated in 1 and 3 weeks, respectively. By testing a series of naturally occurring viruses as well as Delta-Omicron chimeric replicons, we show that Omicron nonstructural protein 6 harbors critical attenuating mutations, which dampen viral RNA replication and reduce lipid droplet consumption. Thus, pGLUE overcomes remaining barriers to broadly study SARS-CoV-2 replication and reveals deficits in nonstructural protein function underlying Omicron attenuation.
]]></description>
<dc:creator>Taha, T. Y.</dc:creator>
<dc:creator>Chen, I. P.</dc:creator>
<dc:creator>Hayashi, J. M.</dc:creator>
<dc:creator>Tabata, T.</dc:creator>
<dc:creator>Walcott, K.</dc:creator>
<dc:creator>Kimmerly, G. R.</dc:creator>
<dc:creator>Syed, A. M.</dc:creator>
<dc:creator>Ciling, A.</dc:creator>
<dc:creator>Suryawanshi, R. K.</dc:creator>
<dc:creator>Martin, H. S.</dc:creator>
<dc:creator>Bach, B. H.</dc:creator>
<dc:creator>Tsou, C.-L.</dc:creator>
<dc:creator>Montano, M.</dc:creator>
<dc:creator>Khalid, M. M.</dc:creator>
<dc:creator>Sreekumar, B. K.</dc:creator>
<dc:creator>Kumar, G. R.</dc:creator>
<dc:creator>Wyman, S.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:date>2023-02-09</dc:date>
<dc:identifier>doi:10.1101/2023.01.31.525914</dc:identifier>
<dc:title><![CDATA[Rapid assembly of SARS-CoV-2 genomes reveals attenuation of the Omicron BA.1 variant through NSP6]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.08.527527v1?rss=1">
<title>
<![CDATA[
Pharmacodynamic model of PARP1 inhibition and global sensitivity analyses can lead to cancer biomarker discovery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.08.527527v1?rss=1</link>
<description><![CDATA[
Pharmacodynamic models provide inroads to understanding key mechanisms of action and may significantly improve patient outcomes in cancer with improved ability to determine therapeutic benefit. Additionally, these models may also lead to insights into potential biomarkers that can be utilized for prediction in prognosis and therapeutic decisions. As an example of this potential, here we present an advanced computational Ordinary Differential Equation (ODE) model of PARP1 signalling and downstream effects due to its inhibition. The model has been validated experimentally and further evaluated through a global sensitivity analysis. The sensitivity analysis uncovered two model parameters related to protein synthesis and degradation rates that were also found to contribute the most variability to the therapeutic prediction. Because this variability may define cancer patient subpopulations, we interrogated genomic, transcriptomic, and clinical databases, to uncover a biomarker that may correspond to patient outcomes in the model. In particular, GSPT2, a GTPase with translation function, was discovered and if mutations serve to alter catalytic activity, its presence may explain the variability in the models parameters. This work offers an analysis of ODE models, inclusive of model development, sensitivity analysis, and ensuing experimental data analysis, and demonstrates the utility of this methodology in uncovering biomarkers in cancer.

Author summaryBecause biochemical reaction networks are complex, dynamic, and typically provide output that results from non-linear interactions, mathematical models of such offer insight into cell function. In the clinic, models including drug action further their usefulness in that they may predict therapeutic outcome and other useful markers such as those for prognosis. In this study, we report a model of drug action that targets a critical protein, that when inhibited, promotes tumor cell death and documented remissions. Because all patients do not respond to the described treatment, a means to find cancer patient subpopulations that might benefit continues to be a challenge. Therefore, we analyzed the pharmacodynamic model by defining the parameters of the greatest variability and interrogated genomic, transcriptomic, and clinical cohort databases with this information and discovered a novel biomarker associated with prognosis in some ovarian and uterine cancer patients and separately, associated with the potential to respond to treatment.
]]></description>
<dc:creator>Mertins, S. D.</dc:creator>
<dc:creator>Isenberg, N. M.</dc:creator>
<dc:creator>Reyes, K.-R.</dc:creator>
<dc:creator>Yoon, B.-J.</dc:creator>
<dc:creator>Urban, N.</dc:creator>
<dc:creator>Jogalekar, M. P.</dc:creator>
<dc:creator>Diolaiti, M. E.</dc:creator>
<dc:creator>Weil, M. R.</dc:creator>
<dc:creator>Stahlberg, E. A.</dc:creator>
<dc:date>2023-02-10</dc:date>
<dc:identifier>doi:10.1101/2023.02.08.527527</dc:identifier>
<dc:title><![CDATA[Pharmacodynamic model of PARP1 inhibition and global sensitivity analyses can lead to cancer biomarker discovery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.09.527892v1?rss=1">
<title>
<![CDATA[
Long COVID manifests with T cell dysregulation, inflammation, and an uncoordinated adaptive immune response to SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.09.527892v1?rss=1</link>
<description><![CDATA[
Long COVID (LC), a type of post-acute sequelae of SARS-CoV-2 infection (PASC), occurs after at least 10% of SARS-CoV-2 infections, yet its etiology remains poorly understood. Here, we used multiple "omics" assays (CyTOF, RNAseq/scRNAseq, Olink) and serology to deeply characterize both global and SARS-CoV-2-specific immunity from blood of individuals with clear LC and non-LC clinical trajectories, 8 months following infection and prior to receipt of any SARS-CoV-2 vaccine. Our analysis focused on deep phenotyping of T cells, which play important roles in immunity against SARS-CoV-2 yet may also contribute to COVID-19 pathogenesis. Our findings demonstrate that individuals with LC exhibit systemic inflammation and immune dysregulation. This is evidenced by global differences in T cell subset distribution in ways that imply ongoing immune responses, as well as by sex-specific perturbations in cytolytic subsets. Individuals with LC harbored increased frequencies of CD4+ T cells poised to migrate to inflamed tissues, and exhausted SARS-CoV-2-specific CD8+ T cells. They also harbored significantly higher levels of SARS-CoV-2 antibodies, and in contrast to non-LC individuals, exhibited a mis-coordination between their SARS-CoV-2-specific T and B cell responses. RNAseq/scRNAseq and Olink analyses similarly revealed immune dysregulatory mechanisms, along with non-immune associated perturbations, in individuals with LC. Collectively, our data suggest that proper crosstalk between the humoral and cellular arms of adaptive immunity has broken down in LC, and that this, perhaps in the context of persistent virus, leads to the immune dysregulation, inflammation, and clinical symptoms associated with this debilitating condition.
]]></description>
<dc:creator>Yin, K.</dc:creator>
<dc:creator>Peluso, M. J.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Shin, M.-G.</dc:creator>
<dc:creator>Neidleman, J.</dc:creator>
<dc:creator>Luo, X.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Anglin, K.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Argueta, U.</dc:creator>
<dc:creator>Lopez, M.</dc:creator>
<dc:creator>Valdivieso, D.</dc:creator>
<dc:creator>Asare, K.</dc:creator>
<dc:creator>Ibrahim, R.</dc:creator>
<dc:creator>Standker, L.</dc:creator>
<dc:creator>Lu, S.</dc:creator>
<dc:creator>Goldberg, S. A.</dc:creator>
<dc:creator>Lee, S. A.</dc:creator>
<dc:creator>Lynch, K. L.</dc:creator>
<dc:creator>Kelly, J. D.</dc:creator>
<dc:creator>Martin, J. N.</dc:creator>
<dc:creator>Muench, J.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Henrich, T. J.</dc:creator>
<dc:creator>Roan, N. R.</dc:creator>
<dc:date>2023-02-10</dc:date>
<dc:identifier>doi:10.1101/2023.02.09.527892</dc:identifier>
<dc:title><![CDATA[Long COVID manifests with T cell dysregulation, inflammation, and an uncoordinated adaptive immune response to SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.09.527927v1?rss=1">
<title>
<![CDATA[
Unified bursting strategies in ectopic and endogenous even-skipped expression patterns 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.09.527927v1?rss=1</link>
<description><![CDATA[
Transcription often occurs in bursts as gene promoters switch stochastically between active and inactive states. Enhancers can dictate transcriptional activity in animal development through the modulation of burst frequency, duration, or amplitude. Previous studies observed that different enhancers can achieve a wide range of transcriptional outputs through the same strategies of bursting control. For example, despite responding to different transcription factors, all even-skipped enhancers increase transcription by upregulating burst frequency and amplitude while burst duration remains largely constant. These shared bursting strategies suggest that a unified molecular mechanism constraints how enhancers modulate transcriptional output. Alternatively, different enhancers could have converged on the same bursting control strategy because of natural selection favoring one of these particular strategies. To distinguish between these two scenarios, we compared transcriptional bursting between endogenous and ectopic gene expression patterns. Because enhancers act under different regulatory inputs in ectopic patterns, dissimilar bursting control strategies between endogenous and ectopic patterns would suggest that enhancers adapted their bursting strategies to their trans-regulatory environment. Here, we generated ectopic even-skipped transcription patterns in fruit fly embryos and discovered that bursting strategies remain consistent in endogenous and ectopic even-skipped expression. These results provide evidence for a unified molecular mechanism shaping even-skipped bursting strategies and serve as a starting point to uncover the realm of strategies employed by other enhancers.
]]></description>
<dc:creator>Berrocal, A.</dc:creator>
<dc:creator>Lammers, N. C.</dc:creator>
<dc:creator>Garcia, H. G.</dc:creator>
<dc:creator>Eisen, M. B.</dc:creator>
<dc:date>2023-02-10</dc:date>
<dc:identifier>doi:10.1101/2023.02.09.527927</dc:identifier>
<dc:title><![CDATA[Unified bursting strategies in ectopic and endogenous even-skipped expression patterns]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.12.528204v1?rss=1">
<title>
<![CDATA[
Integrative single-cell characterization of frugivory adaptations in the bat kidney and pancreas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.12.528204v1?rss=1</link>
<description><![CDATA[
Frugivory evolved multiple times in mammals, including bats. However, the cellular and molecular components driving it remain largely unknown. Here, we used integrative single-cell sequencing on insectivorous and frugivorous bat kidneys and pancreases and identified key cell population, gene expression and regulatory element differences associated with frugivorous adaptation that also relate to human disease, particularly diabetes. We found an increase in collecting duct cells and differentially active genes and regulatory elements involved in fluid and electrolyte balance in the frugivore kidney. In the frugivorous pancreas, we observed an increase in endocrine and a decrease in exocrine cells and differences in genes and regulatory elements involved in insulin regulation. Combined, our work provides novel insights into frugivorous adaptation that also could be leveraged for therapeutic purposes.
]]></description>
<dc:creator>Gordon, W. E.</dc:creator>
<dc:creator>Baek, S.</dc:creator>
<dc:creator>Nguyen, H. P.</dc:creator>
<dc:creator>Kuo, Y.-M.</dc:creator>
<dc:creator>Bradley, R.</dc:creator>
<dc:creator>Galazyuk, A.</dc:creator>
<dc:creator>Lee, I.</dc:creator>
<dc:creator>Ingala, M.</dc:creator>
<dc:creator>Simmons, N. B.</dc:creator>
<dc:creator>Schountz, T.</dc:creator>
<dc:creator>Cooper, L.</dc:creator>
<dc:creator>Georgakopoulos-Soares, I.</dc:creator>
<dc:creator>Hemberg, M.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2023-02-13</dc:date>
<dc:identifier>doi:10.1101/2023.02.12.528204</dc:identifier>
<dc:title><![CDATA[Integrative single-cell characterization of frugivory adaptations in the bat kidney and pancreas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.13.526773v1?rss=1">
<title>
<![CDATA[
Designed Transmembrane Proteins Inhibit the Erythropoietin Receptor in a Custom Binding Topology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.13.526773v1?rss=1</link>
<description><![CDATA[
Transmembrane (TM) domains as simple as a single span can perform complex biological functions using entirely lipid-embedded chemical features. Computational design has potential to generate custom tool molecules directly targeting membrane proteins at their functional TM regions. Thus far, designed TM domain-targeting agents have been limited to mimicking binding modes and motifs of natural TM interaction partners. Here, we demonstrate the design of de novo TM proteins targeting the erythropoietin receptor (EpoR) TM domain in a custom binding topology competitive with receptor homodimerization. The TM proteins expressed in mammalian cells complex with EpoR and inhibit erythropoietin-induced cell proliferation. In vitro, the synthetic TM domain complex outcompetes EpoR homodimerization. Structural characterization reveals that the complex involves the intended amino acids and agrees with our designed molecular model of antiparallel TM helices at 1:1 stoichiometry. Thus, membrane protein TM regions can now be targeted in custom designed topologies.
]]></description>
<dc:creator>Mravic, M.</dc:creator>
<dc:creator>He, L.</dc:creator>
<dc:creator>Kratochvil, H.</dc:creator>
<dc:creator>Hu, H.</dc:creator>
<dc:creator>Nick, S. E.</dc:creator>
<dc:creator>Bai, W.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Jo, H.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>DiMaio, D.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:date>2023-02-15</dc:date>
<dc:identifier>doi:10.1101/2023.02.13.526773</dc:identifier>
<dc:title><![CDATA[Designed Transmembrane Proteins Inhibit the Erythropoietin Receptor in a Custom Binding Topology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.13.528406v1?rss=1">
<title>
<![CDATA[
Transmembrane protein CD69 acts as an S1PR1 agonist 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.13.528406v1?rss=1</link>
<description><![CDATA[
The activation of Sphingosine-1-phosphate receptor 1 (S1PR1) by S1P promotes lymphocyte egress from lymphoid organs, a process critical for immune surveillance and T cell effector activity 1-4. Multiple drugs that inhibit S1PR1 function are in use clinically for the treatment of autoimmune diseases. Cluster of Differentiation 69 (CD69) is an endogenous negative regulator of lymphocyte egress that interacts with S1PR1 in cis to facilitate internalization and degradation of the receptor 5,6. The mechanism by which CD69 causes S1PR1 internalization has been unclear. Moreover, although there are numerous class A GPCR structures determined with different small molecule agonists bound, it remains unknown whether a transmembrane protein per se can act as a class A GPCR agonist. Here, we present the cryo-EM structure of CD69-bound S1PR1 coupled to the heterotrimeric Gi complex. The transmembrane helix (TM) of one protomer of CD69 homodimer contacts the S1PR1-TM4. This interaction allosterically induces the movement of S1PR1-TMs 5-6, directly activating the receptor to engage the heterotrimeric Gi. Mutations in key residues at the interface affect the interactions between CD69 and S1PR1, as well as reduce the receptor internalization. Thus, our structural findings along with functional analyses demonstrate that CD69 acts in cis as a protein agonist of S1PR1, thereby promoting Gi-dependent S1PR1 internalization, loss of S1P gradient sensing, and inhibition of lymphocyte egress.
]]></description>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Qin, Y.</dc:creator>
<dc:creator>Chou, M.</dc:creator>
<dc:creator>Cyster, J. G.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:date>2023-02-15</dc:date>
<dc:identifier>doi:10.1101/2023.02.13.528406</dc:identifier>
<dc:title><![CDATA[Transmembrane protein CD69 acts as an S1PR1 agonist]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.14.528431v1?rss=1">
<title>
<![CDATA[
Insulin-mTOR hyperfunction drives C. elegans aging opposed by the megaprotein LPD-3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.14.528431v1?rss=1</link>
<description><![CDATA[
Insulin-mTOR signaling drives anabolic growth during organismal development, while its late-life dysregulation may detrimentally contribute to aging and limit lifespans. Age-related regulatory mechanisms and functional consequences of insulin-mTOR remain incompletely understood. Here we identify LPD-3 as a megaprotein that orchestrates the tempo of insulin-mTOR signaling during C. elegans aging. We find that an agonist insulin INS-7 is drastically over-produced in early life and shortens lifespan in lpd-3 mutants, a C. elegans model of human Alkuraya-Ku[c]inskas syndrome. LPD-3 forms a bridge-like tunnel megaprotein to facilitate phospholipid trafficking to plasma membranes. Lipidomic profiling reveals increased abundance of hexaceramide species in lpd-3 mutants, accompanied by up-regulation of hexaceramide biosynthetic enzymes, including HYL-1 (Homolog of Yeast Longevity). Reducing HYL-1 activity decreases INS-7 levels and rescues the lifespan of lpd-3 mutants through insulin receptor/DAF-2 and mTOR/LET-363. LPD-3 antagonizes SINH-1, a key mTORC2 component, and decreases expression with age in wild type animals. We propose that LPD-3 acts as a megaprotein brake for aging and its age-dependent decline restricts lifespan through the sphingolipid-hexaceramide and insulin-mTOR pathways.
]]></description>
<dc:creator>Pandey, T.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Zu, J.</dc:creator>
<dc:creator>Deng, H.</dc:creator>
<dc:creator>Shen, K.</dc:creator>
<dc:creator>Vale, G.</dc:creator>
<dc:creator>McDonald, J. G.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2023-02-15</dc:date>
<dc:identifier>doi:10.1101/2023.02.14.528431</dc:identifier>
<dc:title><![CDATA[Insulin-mTOR hyperfunction drives C. elegans aging opposed by the megaprotein LPD-3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.14.528504v1?rss=1">
<title>
<![CDATA[
Tertiary Lymphoid Structures Sustain Cutaneous B cell Activity in Hidradenitis Suppurativa 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.14.528504v1?rss=1</link>
<description><![CDATA[
Structured AbstractO_ST_ABSBackgroundC_ST_ABSHidradenitis suppurativa (HS) skin lesions are highly inflammatory and characterized by a large immune infiltrate. While B cells and plasma cells comprise a major component of this immune milieu the biology and contribution of these cells in HS pathogenesis is unclear.

ObjectiveWe aimed to investigate the dynamics and microenvironmental interactions of B cells within cutaneous HS lesions.

MethodsWe combined histological analysis, single-cell RNA-sequencing (scRNAseq), and spatial transcriptomic profiling of HS lesions to define the tissue microenvironment relative to B cell activity within this disease.

ResultsOur findings identify tertiary lymphoid structures (TLS) within HS lesions and describe organized interactions between T cells, B cells, antigen presenting cells and skin stroma. We find evidence that B cells within HS TLS actively undergo maturation, including participation in germinal center reactions and class switch recombination. Moreover, skin stroma and accumulating T cells are primed to support the formation of TLS and facilitate B cell recruitment during HS.

ConclusionOur data definitively demonstrate the presence of TLS in lesional HS skin and point to ongoing cutaneous B cell maturation through class switch recombination and affinity maturation during disease progression in this inflamed non-lymphoid tissue.
]]></description>
<dc:creator>Lowe, M. M.</dc:creator>
<dc:creator>Cohen, J. N.</dc:creator>
<dc:creator>Moss, M. I.</dc:creator>
<dc:creator>Clancy, S.</dc:creator>
<dc:creator>Adler, J.</dc:creator>
<dc:creator>Yates, A.</dc:creator>
<dc:creator>Naik, H. B.</dc:creator>
<dc:creator>Pauli, M.</dc:creator>
<dc:creator>Taylor, I.</dc:creator>
<dc:creator>McKay, A.</dc:creator>
<dc:creator>Harris, H.</dc:creator>
<dc:creator>Kim, E.</dc:creator>
<dc:creator>Hansen, S. L.</dc:creator>
<dc:creator>Rosenblum, M. D.</dc:creator>
<dc:creator>Moreau, J. M.</dc:creator>
<dc:date>2023-02-15</dc:date>
<dc:identifier>doi:10.1101/2023.02.14.528504</dc:identifier>
<dc:title><![CDATA[Tertiary Lymphoid Structures Sustain Cutaneous B cell Activity in Hidradenitis Suppurativa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.15.528663v1?rss=1">
<title>
<![CDATA[
Massively parallel characterization of psychiatric disorder-associated and cell-type-specific regulatory elements in the developing human cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.15.528663v1?rss=1</link>
<description><![CDATA[
Nucleotide changes in gene regulatory elements are important determinants of neuronal development and disease. Using massively parallel reporter assays in primary human cells from mid-gestation cortex and cerebral organoids, we interrogated the cis-regulatory activity of 102,767 sequences, including differentially accessible cell-type specific regions in the developing cortex and single-nucleotide variants associated with psychiatric disorders. In primary cells, we identified 46,802 active enhancer sequences and 164 disorder-associated variants that significantly alter enhancer activity. Activity was comparable in organoids and primary cells, suggesting that organoids provide an adequate model for the developing cortex. Using deep learning, we decoded the sequence basis and upstream regulators of enhancer activity. This work establishes a comprehensive catalog of functional gene regulatory elements and variants in human neuronal development.

One Sentence SummaryWe identify 46,802 enhancers and 164 psychiatric disorder variants with regulatory effects in the developing cortex and organoids.
]]></description>
<dc:creator>Deng, C.</dc:creator>
<dc:creator>Whalen, S.</dc:creator>
<dc:creator>Steyert, M.</dc:creator>
<dc:creator>Ziffra, R.</dc:creator>
<dc:creator>Przytycki, P. F.</dc:creator>
<dc:creator>Inoue, F.</dc:creator>
<dc:creator>Pereira, D. A.</dc:creator>
<dc:creator>Capauto, D.</dc:creator>
<dc:creator>Norton, S.</dc:creator>
<dc:creator>Vaccarino, F. M.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Ahituv, N. A.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2023-02-15</dc:date>
<dc:identifier>doi:10.1101/2023.02.15.528663</dc:identifier>
<dc:title><![CDATA[Massively parallel characterization of psychiatric disorder-associated and cell-type-specific regulatory elements in the developing human cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.14.528391v1?rss=1">
<title>
<![CDATA[
Generative Molecular Design and Experimental Validation of Selective Histamine H1 Inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.14.528391v1?rss=1</link>
<description><![CDATA[
Generative molecular design (GMD) is an increasingly popular strategy for drug discovery, using machine learning models to propose, evaluate and optimize chemical structures against a set of target design criteria. We present the ATOM-GMD platform, a scalable multiprocessing framework to optimize many parameters simultaneously over large populations of proposed molecules. ATOM-GMD uses a junction tree variational autoencoder mapping structures to latent vectors, along with a genetic algorithm operating on latent vector elements, to search a diverse molecular space for compounds that meet the design criteria. We used the ATOM-GMD framework in a lead optimization case study to develop potent and selective histamine H1 receptor antagonists. We synthesized 103 of the top scoring compounds and measured their properties experimentally. Six of the tested compounds bind H1 with Kis between 10 and 100 nM and are at least 100-fold selective relative to muscarinic M2 receptors, validating the effectiveness of our GMD approach.
]]></description>
<dc:creator>McLoughlin, K. S.</dc:creator>
<dc:creator>Shi, D.</dc:creator>
<dc:creator>Mast, J. E.</dc:creator>
<dc:creator>Bucci, J.</dc:creator>
<dc:creator>Williams, J. P.</dc:creator>
<dc:creator>Jones, W. D.</dc:creator>
<dc:creator>Miyao, D.</dc:creator>
<dc:creator>Nam, L.</dc:creator>
<dc:creator>Osswald, H. L.</dc:creator>
<dc:creator>Zegelman, L.</dc:creator>
<dc:creator>Allen, J. E.</dc:creator>
<dc:creator>Bennion, B. J.</dc:creator>
<dc:creator>Paulson, A. K.</dc:creator>
<dc:creator>Abagyan, R.</dc:creator>
<dc:creator>Head, M. S.</dc:creator>
<dc:creator>Brase, J. M.</dc:creator>
<dc:date>2023-02-16</dc:date>
<dc:identifier>doi:10.1101/2023.02.14.528391</dc:identifier>
<dc:title><![CDATA[Generative Molecular Design and Experimental Validation of Selective Histamine H1 Inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.14.528567v1?rss=1">
<title>
<![CDATA[
Prdm16 and Vcam1 regulate the postnatal disappearance of embryonic radial glia and the ending of cortical neurogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.14.528567v1?rss=1</link>
<description><![CDATA[
Embryonic neural stem cells (NSCs, i.e., radial glia) in the ventricular-subventricular zone (V-SVZ) generate the majority of neurons and glia in the forebrain. Postnatally, embryonic radial glia disappear and a subpopulation of radial glia transition into adult NSCs. As this transition occurs, widespread neurogenesis in brain regions such as the cerebral cortex ends. The mechanisms that regulate the postnatal disappearance of radial glia and the ending of embryonic neurogenesis remain poorly understood. Here, we show that PR domain-containing 16 (Prdm16) promotes the disappearance of radial glia and the ending of neurogenesis in the cerebral cortex. Genetic deletion of Prdm16 from NSCs leads to the persistence of radial glia in the adult V-SVZ and prolonged postnatal cortical neurogenesis. Mechanistically, Prdm16 induces the postnatal reduction in Vascular Cell Adhesion Molecule 1 (Vcam1). The postnatal disappearance of radial glia and the ending of cortical neurogenesis occur normally in Prdm16-Vcam1 double conditional knockout mice. These observations reveal novel molecular regulators of the postnatal disappearance of radial glia and the ending of embryonic neurogenesis, filling a key knowledge gap in NSC biology.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Godoy, M. I.</dc:creator>
<dc:creator>Zhang, A. J.</dc:creator>
<dc:creator>Diamante, G.</dc:creator>
<dc:creator>Ahn, I. S.</dc:creator>
<dc:creator>Cebrian-Silla, A.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Novitch, B.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:date>2023-02-16</dc:date>
<dc:identifier>doi:10.1101/2023.02.14.528567</dc:identifier>
<dc:title><![CDATA[Prdm16 and Vcam1 regulate the postnatal disappearance of embryonic radial glia and the ending of cortical neurogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.15.528738v1?rss=1">
<title>
<![CDATA[
Dual credit assignment processes underliedopamine signals in a complex spatialenvironment. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.15.528738v1?rss=1</link>
<description><![CDATA[
Dopamine in the nucleus accumbens helps motivate behavior based on expectations of future reward ("values"). These values need to be updated by experience: after receiving reward, the choices that led to reward should be assigned greater value. There are multiple theoretical proposals for how this credit assignment could be achieved, but the specific algorithms that generate updated dopamine signals remain uncertain. We monitored accumbens dopamine as freely behaving rats foraged for rewards in a complex, changing environment. We observed brief pulses of dopamine both when rats received reward (scaling with prediction error), and when they encountered novel path opportunities. Furthermore, dopamine ramped up as rats ran towards reward ports, in proportion to the value at each location. By examining the evolution of these dopamine place-value signals, we found evidence for two distinct update processes: progressive propagation along taken paths, as in temporal-difference learning, and inference of value throughout the maze, using internal models. Our results demonstrate that within rich, naturalistic environments dopamine conveys place values that are updated via multiple, complementary learning algorithms.
]]></description>
<dc:creator>Krausz, T. A.</dc:creator>
<dc:creator>Comrie, A. E.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:creator>Daw, N. D.</dc:creator>
<dc:creator>Berke, J. D.</dc:creator>
<dc:date>2023-02-16</dc:date>
<dc:identifier>doi:10.1101/2023.02.15.528738</dc:identifier>
<dc:title><![CDATA[Dual credit assignment processes underliedopamine signals in a complex spatialenvironment.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.15.528761v1?rss=1">
<title>
<![CDATA[
Molecular basis for potent B cell responses to antigen displayed on particles of viral size 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.15.528761v1?rss=1</link>
<description><![CDATA[
Although it has long been appreciated that multivalent antigens - and particularly viral epitope display - produce extremely rapid, robust, and T-independent humoral immune responses, the biochemical basis for such potency has been incompletely understood. Here we take advantage of a set of neutral liposomes of viral size that are engineered to display affinity mutants of the model antigen (Ag) hen egg lysozyme at precisely varied density. We show that particulate Ag display by liposomes induces highly potent B cell responses that are dose-and density-dependent but affinity-independent. Titrating dose of particulate, but not soluble, Ag reveals bimodal Erk phosphorylation and cytosolic calcium increases. Particulate Ag induces signal amplification downstream of the B cell receptor (BCR) by selectively evading LYN-dependent inhibitory pathways, but in vitro potency is independent of CD19. Importantly, Ag display on viral-sized particles signals independently of MYD88 and IRAK1/4, but activates NF-{kappa}B robustly in a manner that mimics T cell help. Together, such biased signaling by particulate Ag promotes MYC expression and reduces the threshold required for B cell proliferation relative to soluble Ag. These findings uncover a molecular basis for highly sensitive B cell response to viral Ag display and remarkable potency of virus-like particle vaccines that is not merely accounted for by avidity and BCR cross-linking, and is independent of the contribution of B cell nucleic acid-sensing machinery.
]]></description>
<dc:creator>Brooks, J. F.</dc:creator>
<dc:creator>Riggs, J.</dc:creator>
<dc:creator>Mueller, J. L.</dc:creator>
<dc:creator>Mathenge, R.</dc:creator>
<dc:creator>Wholey, W.-Y.</dc:creator>
<dc:creator>Yoda, S.-T.</dc:creator>
<dc:creator>Vykunta, V.</dc:creator>
<dc:creator>Cheng, W.</dc:creator>
<dc:creator>Zikherman, J.</dc:creator>
<dc:date>2023-02-16</dc:date>
<dc:identifier>doi:10.1101/2023.02.15.528761</dc:identifier>
<dc:title><![CDATA[Molecular basis for potent B cell responses to antigen displayed on particles of viral size]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.16.528701v1?rss=1">
<title>
<![CDATA[
Redox Regulation of m6A Methyltransferase METTL3 in Human β-cells Controls the Innate Immune Response in Type 1 Diabetes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.16.528701v1?rss=1</link>
<description><![CDATA[
Type 1 Diabetes (T1D) is characterized by autoimmune-mediated destruction of insulin-producing {beta}-cells. Several observations have renewed interest in the innate immune system as an initiator of the disease process against {beta}-cells. Here, we show that N6-Methyladenosine (m6A) is an adaptive {beta}-cell safeguard mechanism that accelerates mRNA decay of the 2-5-oligoadenylate synthetase (OAS) genes to control the antiviral innate immune response at T1D onset. m6A writer methyltransferase 3 (METTL3) levels increase drastically in human and mouse {beta}-cells at T1D onset but rapidly decline with disease progression. Treatment of human islets and EndoC-{beta}H1 cells with pro-inflammatory cytokines interleukin-1 {beta} and interferon  mimicked the METTL3 upregulation seen at T1D onset. Furthermore, m6A-sequencing revealed the m6A hypermethylation of several key innate immune mediators including OAS1, OAS2, and OAS3 in human islets and EndoC-{beta}H1 cells challenged with cytokines. METTL3 silencing in human pseudoislets or EndoC-{beta}H1 cells enhanced OAS levels by increasing its mRNA stability upon cytokine challenge. Consistently, in vivo gene therapy, to prolong Mettl3 overexpression specifically in {beta}-cells, delayed diabetes progression in the non-obese diabetic (NOD) mouse model of T1D by limiting the upregulation of Oas pointing to potential therapeutic relevance. Mechanistically, the accumulation of reactive oxygen species blocked METTL3 upregulation in response to cytokines, while physiological levels of nitric oxide promoted its expression in human islets. Furthermore, for the first time to our knowledge, we show that the cysteines in position C276 and C326 in the zinc finger domain of the METTL3 protein are sensitive to S-nitrosylation (SNO) and are significant for the METTL3 mediated regulation of OAS mRNA stability in human {beta}-cells in response to cytokines. Collectively, we report that m6A regulates human and mouse {beta}-cells to control the innate immune response during the onset of T1D and propose targeting METTL3 to prevent {beta}-cell death in T1D.
]]></description>
<dc:creator>De Jesus, D. F.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Brown, N. K.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Gaffrey, M. J.</dc:creator>
<dc:creator>Kahraman, S.</dc:creator>
<dc:creator>Wei, J.</dc:creator>
<dc:creator>Hu, J.</dc:creator>
<dc:creator>Basile, G.</dc:creator>
<dc:creator>Xiao, L.</dc:creator>
<dc:creator>Rana, T. M.</dc:creator>
<dc:creator>Mathews, C. E.</dc:creator>
<dc:creator>Powers, A. C.</dc:creator>
<dc:creator>Atkinson, M. A.</dc:creator>
<dc:creator>Eizirik, D. L.</dc:creator>
<dc:creator>Dhe-Paganon, S.</dc:creator>
<dc:creator>Parent, A. V.</dc:creator>
<dc:creator>Qian, W.-J.</dc:creator>
<dc:creator>He, C.</dc:creator>
<dc:creator>Kulkarni, R. N.</dc:creator>
<dc:date>2023-02-16</dc:date>
<dc:identifier>doi:10.1101/2023.02.16.528701</dc:identifier>
<dc:title><![CDATA[Redox Regulation of m6A Methyltransferase METTL3 in Human β-cells Controls the Innate Immune Response in Type 1 Diabetes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.16.528876v1?rss=1">
<title>
<![CDATA[
Prospectively predicting BPaMZ Phase IIb outcomes using a translational preclinical mouse to human platform 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.16.528876v1?rss=1</link>
<description><![CDATA[
Despite known treatments, tuberculosis (TB) remains the worlds top infectious killer, highlighting the pressing need for new drug regimens. To prioritize the most efficacious drugs for clinical testing, we previously developed a PK-PD translational platform with bacterial dynamics that reliably predicted short-term monotherapy outcomes in Phase IIa trials from preclinical mouse studies. In this study, we extended our platform to include PK-PD models that account for drug-drug interactions in combination regimens and bacterial regrowth in our bacterial dynamics model to predict cure at end of treatment and relapse 6 months post-treatment. The Phase III trial STAND, testing new regimen pretomanid (Pa), moxifloxacin (M), and pyrazinamide (Z) (PaMZ), predicted to shorten treatment duration by 2 months was put on hold after a separate ongoing trial showed adding bedaquiline (B) to the PaMZ regimen (SimpliciTB) suggested superior efficacy. To forecast if the addition of B would indeed benefit the PaMZ regimen, we applied an extended translational platform to both regimens. We predicted currently available short- and long-term clinical data well for drug combinations related to BPaMZ. We predict the addition of B to PaMZ will shorten treatment duration by 2 months and be non-inferior compared to control HRZE, both at the end of treatment for treatment efficacy and 6 months after treatment has ended in relapse prevention. Using BPaMZ as a case study, we have demonstrated our translational platform can predict Phase II and III outcomes prior to actual trials, allowing us to better prioritize the regimens most likely to succeed.
]]></description>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Goh, J. J. N.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Nuermberger, E.</dc:creator>
<dc:creator>Savic, R. M.</dc:creator>
<dc:date>2023-02-16</dc:date>
<dc:identifier>doi:10.1101/2023.02.16.528876</dc:identifier>
<dc:title><![CDATA[Prospectively predicting BPaMZ Phase IIb outcomes using a translational preclinical mouse to human platform]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.16.528905v1?rss=1">
<title>
<![CDATA[
Broad sensitivity of Candida auris strains to quinolones and mechanisms of resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.16.528905v1?rss=1</link>
<description><![CDATA[
The fungal pathogen Candida auris represents a severe threat to hospitalized patients. Its resistance to multiple classes of antifungal drugs and ability to spread and resist decontamination in health-care settings make it especially dangerous. We screened 1,990 clinically approved and late-stage investigational compounds for the potential to be repurposed as antifungal drugs targeting C. auris and narrowed our focus to five FDA-approved compounds with inhibitory concentrations under 10 {micro}M for C. auris and significantly lower toxicity to three human cell lines. These compounds, some of which had been previously identified in independent screens, include three dihalogenated 8-hydroxyquinolines: broxyquinoline, chloroxine, and clioquinol. A subsequent structure-activity study of 32 quinoline derivatives found that 8-hydroxyquinolines, especially those dihalogenated at the C5 and C7 positions, were the most effective inhibitors of C. auris. To pursue these compounds further, we exposed C. auris to clioquinol in an extended experimental evolution study and found that C. auris developed only 2- to 5-fold resistance to the compound. DNA sequencing of resistant strains and subsequent verification by directed mutation in naive strains revealed that resistance was due to mutations in the transcriptional regulator CAP1 (causing upregulation of the drug transporter MDR1) and in the drug transporter CDR1. These mutations had only modest effects on resistance to traditional antifungal agents, and the CDR1 mutation rendered C. auris more sensitive to posaconazole. This observation raises the possibility that a combination treatment involving an 8-hydroxyquinoline and posaconazole might prevent C. auris from developing resistance to this established antifungal agent.

Abstract ImportanceThe rapidly emerging fungal pathogen Candida auris represents a growing threat to hospitalized patients, in part due to frequent resistance to multiple classes of antifungal drugs. We identify a class of compounds, the dihalogenated hydroxyquinolines, with broad fungistatic ability against a diverse collection of 13 strains of C. auris. Although this compound has been identified in previous screens, we extended the analysis by showing that C. auris developed only modest 2- to 5-fold increases in resistance to this class of compounds despite long-term exposure; a noticeable difference from the 30- to 500- fold increases in resistance reported for similar studies with commonly used antifungal drugs. We also identify the mutations underlying the resistance. These results suggest that the dihalogenated hydroxyquinolines are working inside the fungal cell and should be developed further to combat C. auris and other fungal pathogens.

TweetLohse and colleagues characterize a class of compounds that inhibit the fungal pathogen C. auris. Unlike many other antifungal drugs, C. auris does not readily develop resistance to this class of compounds.
]]></description>
<dc:creator>Lohse, M. B.</dc:creator>
<dc:creator>Laurie, M. T.</dc:creator>
<dc:creator>Levan, S.</dc:creator>
<dc:creator>Ziv, N.</dc:creator>
<dc:creator>Ennis, C. L.</dc:creator>
<dc:creator>Nobile, C. J.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Johnson, A. D.</dc:creator>
<dc:date>2023-02-17</dc:date>
<dc:identifier>doi:10.1101/2023.02.16.528905</dc:identifier>
<dc:title><![CDATA[Broad sensitivity of Candida auris strains to quinolones and mechanisms of resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.20.528987v1?rss=1">
<title>
<![CDATA[
DeepHeme: A generalizable, bone marrow classifierwith hematopathologist-level performance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.20.528987v1?rss=1</link>
<description><![CDATA[
Morphology-based classification of cells in the bone marrow aspirate (BMA) is a key step in the diagnosis and management of hematologic malignancies. However, it is time-intensive and must be performed by expert hematopathologists and laboratory professionals. We curated a large, high-quality dataset of 41,595 hematopathologist consensus-annotated single-cell images extracted from BMA whole slide images (WSIs) containing 23 morphologic classes from the clinical archives of the University of California, San Francisco. We trained a convolutional neural network, DeepHeme, to classify images in this dataset, achieving a mean area under the curve (AUC) of 0.99. DeepHeme was then externally validated on WSIs from Memorial Sloan Kettering Cancer Center, with a similar AUC of 0.98, demonstrating robust generalization. When compared to individual hematopathologists from three different top academic medical centers, the algorithm outperformed all three. Finally, DeepHeme reliably identified cell states such as mitosis, paving the way for image-based quantification of mitotic index in a cell-specific manner, which may have important clinical applications.
]]></description>
<dc:creator>Goldgof, G.</dc:creator>
<dc:creator>Sun, S.</dc:creator>
<dc:creator>Cleaves, J.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Lucas, F.</dc:creator>
<dc:creator>Brown, L.</dc:creator>
<dc:creator>Spectors, J.</dc:creator>
<dc:creator>Boiocchi, L.</dc:creator>
<dc:creator>Baik, J.</dc:creator>
<dc:creator>Zhu, M.</dc:creator>
<dc:creator>Ardon, O.</dc:creator>
<dc:creator>Lu, C.</dc:creator>
<dc:creator>Dogan, A.</dc:creator>
<dc:creator>Goldgof, D.</dc:creator>
<dc:creator>Carmichael, I.</dc:creator>
<dc:creator>Prakash, S.</dc:creator>
<dc:creator>Butte, A.</dc:creator>
<dc:date>2023-02-21</dc:date>
<dc:identifier>doi:10.1101/2023.02.20.528987</dc:identifier>
<dc:title><![CDATA[DeepHeme: A generalizable, bone marrow classifierwith hematopathologist-level performance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.20.529175v1?rss=1">
<title>
<![CDATA[
Tracing cancer evolution and heterogeneity using Hi-C 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.20.529175v1?rss=1</link>
<description><![CDATA[
Chromosomal rearrangements can initiate and drive cancer progression, yet it has been challenging to evaluate their impact, especially in genetically heterogeneous solid cancers. To address this problem we developed HiDENSEC, a new computational framework for analyzing chromatin conformation capture in heterogeneous samples, which can infer somatic copy number alterations, characterize large-scale chromosomal rearrangements, and estimate cancer cell fractions. We validated HiDENSEC with in silico and in vitro controls, and then characterized chromosome-scale evolution during melanoma progression in formalin-fixed tumor samples from three patients. The resulting comprehensive annotation of the genomic events includes copy number neutral translocations that disrupt tumor suppressor genes such as NF1, whole chromosome arm exchanges that result in loss of CDKN2A, and whole-arm copy-number neutral loss of homozygosity involving PTEN. These findings show that large-scale chromosomal rearrangements occur throughout cancer evolution and characterizing these events yields insights into drivers of melanoma progression.
]]></description>
<dc:creator>Erdmann-Pham, D. D.</dc:creator>
<dc:creator>Batra, S. S.</dc:creator>
<dc:creator>Turkalo, T. K.</dc:creator>
<dc:creator>Durbin, J.</dc:creator>
<dc:creator>Blanchette, M.</dc:creator>
<dc:creator>Yeh, I.</dc:creator>
<dc:creator>Shain, H.</dc:creator>
<dc:creator>Bastian, B.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Rokhsar, D. S.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:date>2023-02-21</dc:date>
<dc:identifier>doi:10.1101/2023.02.20.529175</dc:identifier>
<dc:title><![CDATA[Tracing cancer evolution and heterogeneity using Hi-C]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.21.529426v1?rss=1">
<title>
<![CDATA[
TET1 Catalytic Activity is Required for Reprogramming of Imprinting Control Regions and Patterning of Sperm-Specific Hypomethylated Regions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.21.529426v1?rss=1</link>
<description><![CDATA[
DNA methylation erasure is required for mammalian primordial germ cell reprogramming. TET enzymes iteratively oxidize 5-methylcytosine to generate 5-hyroxymethylcytosine (5hmC), 5-formylcytosine, and 5-carboxycytosine to facilitate active genome demethylation. Whether these bases are required to promote replication-coupled dilution or activate base excision repair during germline reprogramming remains unresolved due to the lack of genetic models that decouple TET activities. Here, we generated two mouse lines expressing catalytically inactive TET1 (Tet1-HxD) and TET1 that stalls oxidation at 5hmC (Tet1-V). Tet1-/-, Tet1V/V, and Tet1HxD/HxD sperm methylomes show that TET1V and TET1HxD rescue most Tet1-/- hypermethylated regions, demonstrating the importance of TET1s extra-catalytic functions. Imprinted regions, in contrast, require iterative oxidation. We further reveal a broader class of hypermethylated regions in sperm of Tet1 mutant mice that are excluded from de novo methylation during male germline development and depend on TET oxidation for reprogramming. Our study underscores the link between TET1-mediated demethylation during reprogramming and sperm methylome patterning.
]]></description>
<dc:creator>Prasasya, R. D.</dc:creator>
<dc:creator>Caldwell, B. A.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Wu, S.</dc:creator>
<dc:creator>Leu, N. A. D.</dc:creator>
<dc:creator>Fowler, J. M.</dc:creator>
<dc:creator>Cincotta, S. A.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:creator>Kohli, R. M.</dc:creator>
<dc:creator>Bartolomei, M.</dc:creator>
<dc:date>2023-02-21</dc:date>
<dc:identifier>doi:10.1101/2023.02.21.529426</dc:identifier>
<dc:title><![CDATA[TET1 Catalytic Activity is Required for Reprogramming of Imprinting Control Regions and Patterning of Sperm-Specific Hypomethylated Regions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529616v1?rss=1">
<title>
<![CDATA[
MAP9/MAPH-9 supports axonemal microtubule doublets and modulates motor movement 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529616v1?rss=1</link>
<description><![CDATA[
Microtubule doublets (MTDs) are a well conserved compound microtubule structure found primarily in cilia. However, the mechanisms by which MTDs form and are maintained in vivo remain poorly understood. Here, we characterize microtubule-associated protein 9 (MAP9) as a novel MTD-associated protein. We demonstrate that C. elegans MAPH-9, a MAP9 homolog, is present during MTD assembly and localizes exclusively to MTDs, a preference that is in part mediated by tubulin polyglutamylation. Loss of MAPH-9 caused ultrastructural MTD defects, dysregulated axonemal motor velocity, and perturbed cilia function. As we found that the mammalian ortholog MAP9 localized to axonemes in cultured mammalian cells and mouse tissues, we propose that MAP9/MAPH-9 plays a conserved role in supporting the structure of axonemal MTDs and regulating ciliary motors.
]]></description>
<dc:creator>Tran, M. V.</dc:creator>
<dc:creator>Ferguson, J. W.</dc:creator>
<dc:creator>Cote, L. E.</dc:creator>
<dc:creator>Khuntsariya, D.</dc:creator>
<dc:creator>Fetter, R. D.</dc:creator>
<dc:creator>Wang, J. T.</dc:creator>
<dc:creator>Wellard, S. R.</dc:creator>
<dc:creator>Sallee, M. D.</dc:creator>
<dc:creator>Eskinazi, S.</dc:creator>
<dc:creator>Genova, M.</dc:creator>
<dc:creator>Magiera, M. M.</dc:creator>
<dc:creator>Janke, C.</dc:creator>
<dc:creator>Stearns, T.</dc:creator>
<dc:creator>Lansky, Z.</dc:creator>
<dc:creator>Shen, K.</dc:creator>
<dc:creator>Magescas, J.</dc:creator>
<dc:creator>Feldman, J. L.</dc:creator>
<dc:date>2023-02-23</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529616</dc:identifier>
<dc:title><![CDATA[MAP9/MAPH-9 supports axonemal microtubule doublets and modulates motor movement]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529756v1?rss=1">
<title>
<![CDATA[
Candida albicans stimulates the formation of a multi-receptor complex that mediates epithelial cell invasion during oropharyngeal infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529756v1?rss=1</link>
<description><![CDATA[
Fungal invasion of the oral epithelium is central to the pathogenesis of oropharyngeal candidiasis (OPC). Candida albicans invades the oral epithelium by receptor-induced endocytosis but this process is incompletely understood. We found that C. albicans infection of oral epithelial cells induces c-Met to form a multi-protein complex with E-cadherin and the epidermal growth factor receptor (EGFR). E-cadherin is necessary for C. albicans to activate both c-Met and EGFR and to induce the endocytosis of C. albicans. Proteomics analysis revealed that c-Met interacts with C. albicans Hyr1, Als3 and Ssa1. Both Hyr1 and Als3 were required for C. albicans stimulation of c-Met and EGFR in oral epithelial cells in vitro and for full virulence during OPC in mice. Treating mice with small molecule inhibitors of c-Met and EGFR ameliorated OPC, demonstrating the potential therapeutic efficacy of blocking these host receptors for C. albicans.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=161 HEIGHT=200 SRC="FIGDIR/small/529756v1_ufig1.gif" ALT="Figure 1">
View larger version (30K):
org.highwire.dtl.DTLVardef@1448372org.highwire.dtl.DTLVardef@111b411org.highwire.dtl.DTLVardef@144f3b8org.highwire.dtl.DTLVardef@1dcf548_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LIc-Met is an oral epithelial cell receptor for Candida albicans
C_LIO_LIC. albicans infection causes c-Met and the epidermal growth factor receptor (EGFR) to form a complex with E-cadherin, which is required for c-Met and EGFR function
C_LIO_LIC. albicans Hyr1 and Als3 interact with c-Met and EGFR, inducing oral epithelial cell endocytosis and virulence during oropharyngeal candidiasis
C_LIO_LIDual blockade of c-Met and EGFR ameliorates oropharyngeal candidiasis
C_LI
]]></description>
<dc:creator>Phan, Q. T.</dc:creator>
<dc:creator>Solis, N. V.</dc:creator>
<dc:creator>Cravener, M. V.</dc:creator>
<dc:creator>Swidergall, M.</dc:creator>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Huang, M. Y.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Ibrahim, A. S.</dc:creator>
<dc:creator>Mazzone, M.</dc:creator>
<dc:creator>Mitchell, A. P.</dc:creator>
<dc:creator>Filler, S. G.</dc:creator>
<dc:date>2023-02-23</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529756</dc:identifier>
<dc:title><![CDATA[Candida albicans stimulates the formation of a multi-receptor complex that mediates epithelial cell invasion during oropharyngeal infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529575v1?rss=1">
<title>
<![CDATA[
Multi-omic screening of invasive GBM cells in engineered biomaterials and patient biopsies reveals targetable transsulfuration pathway alterations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529575v1?rss=1</link>
<description><![CDATA[
While the poor prognosis of glioblastoma arises from the invasion of a subset of tumor cells, little is known of the metabolic alterations within these cells that fuel invasion. We integrated spatially addressable hydrogel biomaterial platforms, patient site-directed biopsies, and multi-omics analyses to define metabolic drivers of invasive glioblastoma cells. Metabolomics and lipidomics revealed elevations in the redox buffers cystathionine, hexosylceramides, and glucosyl ceramides in the invasive front of both hydrogel-cultured tumors and patient site-directed biopsies, with immunofluorescence indicating elevated reactive oxygen species (ROS) markers in invasive cells. Transcriptomics confirmed upregulation of ROS-producing and response genes at the invasive front in both hydrogel models and patient tumors. Amongst oncologic ROS, hydrogen peroxide specifically promoted glioblastoma invasion in 3D hydrogel spheroid cultures. A CRISPR metabolic gene screen revealed cystathionine gamma lyase (CTH), which converts cystathionine to the non-essential amino acid cysteine in the transsulfuration pathway, to be essential for glioblastoma invasion. Correspondingly, supplementing CTH knockdown cells with exogenous cysteine rescued invasion. Pharmacologic CTH inhibition suppressed glioblastoma invasion, while CTH knockdown slowed glioblastoma invasion in vivo. Our studies highlight the importance of ROS metabolism in invasive glioblastoma cells and support further exploration of the transsulfuration pathway as a mechanistic and therapeutic target.
]]></description>
<dc:creator>Garcia, J. H.</dc:creator>
<dc:creator>Akins, E. A.</dc:creator>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Wolf, K. J.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Choudhary, N.</dc:creator>
<dc:creator>Lad, M.</dc:creator>
<dc:creator>Shukla, P.</dc:creator>
<dc:creator>Gill, S.</dc:creator>
<dc:creator>Carson, W.</dc:creator>
<dc:creator>Carette, L.</dc:creator>
<dc:creator>Zheng, A.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:date>2023-02-24</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529575</dc:identifier>
<dc:title><![CDATA[Multi-omic screening of invasive GBM cells in engineered biomaterials and patient biopsies reveals targetable transsulfuration pathway alterations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529747v1?rss=1">
<title>
<![CDATA[
Vagus nerve stimulation using an endovascular electrode array 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529747v1?rss=1</link>
<description><![CDATA[
Vagus nerve stimulation (VNS), which involves a surgical procedure to place electrodes directly on the vagus nerve (VN), is approved clinically for the treatment of epilepsy, depression, and to facilitate rehabilitation in stroke. VNS at surgically implanted electrodes is often limited by activation of motor nerve fibers near and within the VN that cause neck muscle contraction. In this study we investigated endovascular VNS that may allow activation of the VN at locations where the motor nerve fibers are not localized. We used endovascular electrodes within the nearby internal jugular vein (IJV) to electrically stimulate the VN while recording VN compound action potentials and neck muscle motor evoked potentials in an acute intraoperative swine experiment. We show that the stimulation electrode position within the IJV is critical for efficient activation of the VN. We also demonstrate use of fluoroscopy (cone beam CT mode) and ultrasound to determine the position of the endovascular stimulation electrode with respect to the VN and IJV. At the most effective endovascular stimulation locations tested, thresholds for VN activation were several times higher than direct stimulation of the nerve using a cuff electrode; however, this work demonstrates the feasibility of VNS with endovascular electrodes and provides tools to optimize endovascular electrode positions for VNS. The ability to stimulate the VN with endovascular electrodes creates the possibility to stimulate at VN locations that would be otherwise too invasive and at VN locations where structures such as motor nerve fibers do not exist.
]]></description>
<dc:creator>Nicolai, E. N.</dc:creator>
<dc:creator>Arturo Larco, J.</dc:creator>
<dc:creator>Madhani, S. I.</dc:creator>
<dc:creator>Asirvatham, S. J.</dc:creator>
<dc:creator>Chang, S.-y.</dc:creator>
<dc:creator>Ludwig, K. A.</dc:creator>
<dc:creator>Savastano, L. E.</dc:creator>
<dc:creator>Worrell, G. A.</dc:creator>
<dc:date>2023-02-24</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529747</dc:identifier>
<dc:title><![CDATA[Vagus nerve stimulation using an endovascular electrode array]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.24.529917v1?rss=1">
<title>
<![CDATA[
Fate of telomere entanglements is dictated by the timing of anaphase midregion nuclear envelope breakdown 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.24.529917v1?rss=1</link>
<description><![CDATA[
Persisting replication intermediates can confer mitotic catastrophe if left unresolved. Loss of the fission yeast telomere protein Taz1 (ortholog of mammalian TRF1/TRF2) causes telomeric replication fork stalling and in turn, telomere entanglements that stretch between the segregating chromosomes at anaphase. At [&le;]20{degrees}C, these entanglements fail to resolve, resulting in lethality. Rif1, a conserved DNA replication/repair protein, localizes between the segregating chromosomes and specifically hinders the resolution of telomere entanglements without affecting their formation. During anaphase, the spindle and the last segments of segregating chromatin are encased by the nuclear envelope (NE), creating a microdomain termed the anaphase midregion. In the final stages of fission yeast mitosis, this midregion undergoes local NE breakdown. Here we demonstrate that in response to taz1{Delta} telomeric entanglements, Rif1 delays midregion NE breakdown, and this delay disfavors entanglement resolution. Accordingly, gene deletions that hasten midregion NE breakdown phenocopy, and are epistatic with, rif1+ deletion. Conversely, gene deletions that delay midregion NE breakdown block the taz1{Delta} telomere detanglement afforded by loss of Rif1. Overexpression of Rif1 in a wild type background causes cold-specific NE defects and lethality, which are rescued by treatment with a membrane fluidizing agent. We propose that delayed NE breakdown normally favors the resolution of simple entanglements, arising from incomplete replication, by delaying exposure to the cytoplasm. In contrast, resolution of more complex entanglements involving strand invasion, like entanglements between nonsister taz1{Delta} telomeres, requires more rapid exposure to the cytoplasm. These observations uncover an unexpected coordination between NE remodeling and DNA processing events that can prevent or promote aneuploidy.
]]></description>
<dc:creator>Nageshan, R. K.</dc:creator>
<dc:creator>Ortega, R.</dc:creator>
<dc:creator>Krogan, N.</dc:creator>
<dc:creator>Cooper, J. P.</dc:creator>
<dc:date>2023-02-25</dc:date>
<dc:identifier>doi:10.1101/2023.02.24.529917</dc:identifier>
<dc:title><![CDATA[Fate of telomere entanglements is dictated by the timing of anaphase midregion nuclear envelope breakdown]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529813v1?rss=1">
<title>
<![CDATA[
Dynamic functional connectivity MEG features of Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529813v1?rss=1</link>
<description><![CDATA[
Dynamic resting state functional connectivity (RSFC) characterizes time-varying fluctuations of functional brain network activity. While many studies have investigated static functional connectivity, it has been unclear whether features of dynamic functional connectivity are associated with neurodegenerative diseases. Popular sliding-window and clustering methods for extracting dynamic RSFC have various limitations that prevent extracting reliable features to address this question. Here, we use a novel and robust time-varying dynamic network (TVDN) approach to extract the dynamic RSFC features from high resolution magnetoencephalography (MEG) data of participants with Alzheimers disease (AD) and matched controls. The TVDN algorithm automatically and adaptively learns the low-dimensional spatiotemporal manifold of dynamic RSFC and detects dynamic state transitions in data. We show that amongst all the functional features we investigated, the dynamic manifold features are the most predictive of AD. These include: the temporal complexity of the brain network, given by the number of state transitions and their dwell times, and the spatial complexity of the brain network, given by the number of eigenmodes. These dynamic features have high sensitivity and specificity in distinguishing AD from healthy subjects. Intriguingly, we found that AD patients generally have higher spatial complexity but lower temporal complexity compared with healthy controls. We also show that graph theoretic metrics of dynamic component of TVDN are significantly different in AD versus controls, while static graph metrics are not statistically different. These results indicate that dynamic RSFC features are impacted in neurodegenerative disease like Alzheimers disease, and may be crucial to understanding the pathophysiological trajectory of these diseases.
]]></description>
<dc:creator>Jin, H.</dc:creator>
<dc:creator>Ranasinghe, K.</dc:creator>
<dc:creator>Prahu, P.</dc:creator>
<dc:creator>Dale, C.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Kudo, K.</dc:creator>
<dc:creator>Vossel, K.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Jiang, F.</dc:creator>
<dc:date>2023-02-26</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529813</dc:identifier>
<dc:title><![CDATA[Dynamic functional connectivity MEG features of Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.24.529939v1?rss=1">
<title>
<![CDATA[
Idioblasts accumulating anticancer alkaloids in Catharanthus roseus leaves are a unique cell type 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.24.529939v1?rss=1</link>
<description><![CDATA[
Catharanthus roseus leaves produce a range of monoterpenoid indole alkaloids (MIAs) that include low levels of the anticancer drugs vinblastine and vincristine. The MIA pathway displays a complex architecture spanning different subcellular and cell-type localizations and is under complex regulation. As a result, the development of strategies to increase the levels of the anticancer MIAs has remained elusive. The pathway involves mesophyll specialised idioblasts where the late unsolved biosynthetic steps are thought to occur. Here, protoplasts of C. roseus leaf idioblasts were isolated by fluorescence-activated cell sorting, and their differential alkaloid and transcriptomic profiles were characterised. This involved the assembly of an improved C. roseus transcriptome from short- and long-read data, IDIO+. It was observed that C. roseus mesophyll idioblasts possess a distinctive transcriptomic profile associated with protection against biotic and abiotic stresses, and indicative that this cell type is a carbon sink, in contrast with surrounding mesophyll cells. Moreover, it is shown that idioblasts are a hotspot of alkaloid accumulation, suggesting that their transcriptome may hold the keys to the in-depth understanding of the MIA pathway and the success of strategies leading to higher levels of the anticancer drugs.

HighlightCatharanthus mesophyll idioblasts are a hotspot of anticancer alkaloid accumulation. The idioblast transcriptome reveals commitment with stress responses and provides a roadmap towards the increase of anticancer alkaloid levels.
]]></description>
<dc:creator>Guedes, J. G.</dc:creator>
<dc:creator>Ribeiro, R.</dc:creator>
<dc:creator>Carqueijeiro, I.</dc:creator>
<dc:creator>Guimaraes, A. L.</dc:creator>
<dc:creator>Bispo, C.</dc:creator>
<dc:creator>Archer, J.</dc:creator>
<dc:creator>Azevedo, H.</dc:creator>
<dc:creator>Fonseca, N. A.</dc:creator>
<dc:creator>Sottomayor, M.</dc:creator>
<dc:date>2023-02-27</dc:date>
<dc:identifier>doi:10.1101/2023.02.24.529939</dc:identifier>
<dc:title><![CDATA[Idioblasts accumulating anticancer alkaloids in Catharanthus roseus leaves are a unique cell type]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.26.530135v1?rss=1">
<title>
<![CDATA[
Acetylcholine engages distinct amygdala microcircuits to gate internal theta rhythm 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.26.530135v1?rss=1</link>
<description><![CDATA[
Acetylcholine (ACh) is released from basal forebrain cholinergic neurons in response to salient stimuli and engages brain states supporting attention and memory. These high ACh states are associated with theta oscillations, which synchronize neuronal ensembles. Theta oscillations in basolateral amygdala (BLA) underlie emotional memory, yet their mechanism remains unclear. Using brain slice electrophysiology in mice, we show large ACh stimuli evoke prolonged theta oscillations in BLA local field potential that depend upon activation of cholecystokinin (CCK) interneurons (INs). Somatostatin (SOM) INs inhibit CCK INs and are themselves inhibited by ACh, gating BLA theta. ACh-induced theta activity is more readily evoked in BLA over cortex or hippocampus, suggesting preferential activation of BLA during high ACh states. These data reveal a SOM-CCK IN circuit in BLA that gates internal theta oscillations and suggest a mechanism by which salient stimuli acting through ACh switch the BLA into a network state enabling emotional memory.
]]></description>
<dc:creator>Bratsch-Prince, J. X.</dc:creator>
<dc:creator>Warren, J. W.</dc:creator>
<dc:creator>Jones, G. C.</dc:creator>
<dc:creator>McDonald, A. J.</dc:creator>
<dc:creator>Mott, D. D.</dc:creator>
<dc:date>2023-02-27</dc:date>
<dc:identifier>doi:10.1101/2023.02.26.530135</dc:identifier>
<dc:title><![CDATA[Acetylcholine engages distinct amygdala microcircuits to gate internal theta rhythm]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.27.529682v1?rss=1">
<title>
<![CDATA[
Uncovering the essential roles of human GCP 2 orthologs in Caenorhabditis elegans. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.27.529682v1?rss=1</link>
<description><![CDATA[
Human glutamate carboxypeptidase 2 (GCP2) from the M28B metalloprotease group is an important target for therapy in neurological disorders and an established tumor marker. However, its physiological functions remain unclear. To better understand general roles, we used the model organism Caenorhabditis elegans genetically manipulate its three existing orthologous genes and evaluate the impact on worm physiology. The results of gene knockout studies showed that C. elegans GCP2 orthologs affect the pharyngeal physiology, reproduction, and structural integrity of the organism. Promoter-driven GFP expression revealed distinct localization for each of the three gene paralogs, with gcp-2.1 being most abundant in muscles, intestine, and pharyngeal interneurons, gcp-2.2 restricted to the phasmid neurons, and gcp-2.3 located in the excretory cell. This study provides new insight into the unique phenotypic effects of GCP2 gene knockouts in C. elegans, and the specific tissue localizations. We believe that elucidation of particular roles in a non-mammalian organism can help to explain important questions linked to human GCP2 physiology and in extension to GCP2 involvement in pathophysiological processes.
]]></description>
<dc:creator>Panska, L.</dc:creator>
<dc:creator>Nedvedova, S.</dc:creator>
<dc:creator>Vacek, V.</dc:creator>
<dc:creator>Krivska, D.</dc:creator>
<dc:creator>Konecny, L.</dc:creator>
<dc:creator>Knop, F.</dc:creator>
<dc:creator>Kutil, Z.</dc:creator>
<dc:creator>Skultetyova, L.</dc:creator>
<dc:creator>Leontovyc, A.</dc:creator>
<dc:creator>Ulrychova, L.</dc:creator>
<dc:creator>Sakanari, J.</dc:creator>
<dc:creator>Asahina, M.</dc:creator>
<dc:creator>Barinka, C.</dc:creator>
<dc:creator>Macurkova, M.</dc:creator>
<dc:creator>Dvorak, J.</dc:creator>
<dc:date>2023-02-27</dc:date>
<dc:identifier>doi:10.1101/2023.02.27.529682</dc:identifier>
<dc:title><![CDATA[Uncovering the essential roles of human GCP 2 orthologs in Caenorhabditis elegans.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.27.530345v1?rss=1">
<title>
<![CDATA[
Systematic Identification of Post-Transcriptional Regulatory Modules 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.27.530345v1?rss=1</link>
<description><![CDATA[
In our cells, a limited number of RNA binding proteins (RBPs) are responsible for all aspects of RNA metabolism across the entire transcriptome. To accomplish this, RBPs form regulatory units that act on specific target regulons. However, the landscape of RBP combinatorial interactions remains poorly explored. Here, we performed a systematic annotation of RBP combinatorial interactions via multimodal data integration. We built a large-scale map of RBP protein neighborhoods by generating in vivo proximity-dependent biotinylation datasets of 50 human RBPs. In parallel, we used CRISPR interference with single-cell readout to capture transcriptomic changes upon RBP knockdowns. By combining these physical and functional interaction readouts, along with the atlas of RBP mRNA targets from eCLIP assays, we generated an integrated map of functional RBP interactions. We then used this map to match RBPs to their context-specific functions and validated the predicted functions biochemically for four RBPs. This study highlights the previously underappreciated scale of the inter-RBP interactions, be it genetic or physical, and is a first step towards a more comprehensive understanding of post-transcriptional regulatory processes and their underlying molecular grammar.
]]></description>
<dc:creator>Khoroshkin, M. S.</dc:creator>
<dc:creator>Buyan, A.</dc:creator>
<dc:creator>Dodel, M.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Trejo, F.</dc:creator>
<dc:creator>Doty, A.</dc:creator>
<dc:creator>Baratam, R.</dc:creator>
<dc:creator>Zhou, S.</dc:creator>
<dc:creator>Joshi, T.</dc:creator>
<dc:creator>Miglani, S.</dc:creator>
<dc:creator>Choi, B.</dc:creator>
<dc:creator>Subramanyam, V.</dc:creator>
<dc:creator>Modi, H.</dc:creator>
<dc:creator>Corces, R.</dc:creator>
<dc:creator>Markett, D.</dc:creator>
<dc:creator>Kulakovskiy, I. V.</dc:creator>
<dc:creator>Mardakheh, F. K.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2023-03-01</dc:date>
<dc:identifier>doi:10.1101/2023.02.27.530345</dc:identifier>
<dc:title><![CDATA[Systematic Identification of Post-Transcriptional Regulatory Modules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.28.530341v1?rss=1">
<title>
<![CDATA[
Far From Home: Evolution of haploid and diploid populations reveals common, strong, and variable pleiotropic effects in non-home environments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.28.530341v1?rss=1</link>
<description><![CDATA[
Adaptation is driven by the selection for beneficial mutations that provide a fitness advantage in the specific environment in which a population is evolving. However, environments are rarely constant or predictable. When an organism well adapted to one environment finds itself in another, pleiotropic effects of mutations that made it well adapted to its former environment will affect its success. To better understand such pleiotropic effects, we evolved both haploid and diploid barcoded budding yeast populations in multiple environments, isolated adaptive clones, and then determined the fitness effects of adaptive mutations in "non-home" environments in which they were not selected. We find that pleiotropy is common, with most adaptive evolved lineages showing fitness effects in non-home environments. Consistent with other studies, we find that these pleiotropic effects are unpredictable: they are beneficial in some environments and deleterious in others. However, we do find that lineages with adaptive mutations in the same genes tend to show similar pleiotropic effects. We also find that ploidy influences the observed adaptive mutational spectra in a condition-specific fashion. In some conditions, haploids and diploids are selected with adaptive mutations in identical genes, while in others they accumulate mutations in almost completely disjoint sets of genes.
]]></description>
<dc:creator>Chen, V. K.</dc:creator>
<dc:creator>Johnson, M. S.</dc:creator>
<dc:creator>Herissant, L.</dc:creator>
<dc:creator>Humphrey, P. T.</dc:creator>
<dc:creator>Yuan, D. C.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Agarwala, A.</dc:creator>
<dc:creator>Hoelscher, S. B.</dc:creator>
<dc:creator>Petrov, D.</dc:creator>
<dc:creator>Desai, M. M.</dc:creator>
<dc:creator>Sherlock, G. J.</dc:creator>
<dc:date>2023-03-01</dc:date>
<dc:identifier>doi:10.1101/2023.02.28.530341</dc:identifier>
<dc:title><![CDATA[Far From Home: Evolution of haploid and diploid populations reveals common, strong, and variable pleiotropic effects in non-home environments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.01.530698v1?rss=1">
<title>
<![CDATA[
Inhibitory control of speech production in the human premotor frontal cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.01.530698v1?rss=1</link>
<description><![CDATA[
Natural speech is full of starts and stops. Here, we studied the neural mechanisms that underlie the inhibitory control of speech, specifically the ability to stop speaking on demand. We recorded direct cortical activity while participants made continuous speech production and were given a visual cue to stop speaking. Neural recordings revealed activity in the premotor frontal cortex associated with speech stopping. Cortical sites showing stop activity were largely distinct from sites involved in active speech production or, more specifically, encoding articulatory movements. Electrocortical stimulation mapping at many premotor sites with stop activity caused involuntary speech arrest, an immediate inability to speak or vocalize. Furthermore, many speech arrest sites did not co-localize with neural activity correlating with speech motor planning or execution, contrary to this long-assumed function in clinical brain mapping. Together, these results suggest a previously unknown premotor cortical network that underlies the inhibitory control of speech, which has significant implications for understanding the dynamics of normal and altered speech production, as well as clinical brain mapping.
]]></description>
<dc:creator>Zhao, L.</dc:creator>
<dc:creator>Silva, A. B.</dc:creator>
<dc:creator>Kurteff, G. L.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:date>2023-03-01</dc:date>
<dc:identifier>doi:10.1101/2023.03.01.530698</dc:identifier>
<dc:title><![CDATA[Inhibitory control of speech production in the human premotor frontal cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.01.530704v1?rss=1">
<title>
<![CDATA[
Bayesian Inference of a Spectral Graph Model for Brain Oscillations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.01.530704v1?rss=1</link>
<description><![CDATA[
The relationship between brain functional connectivity and structural connectivity has caught extensive attention of the neuroscience community, commonly inferred using mathematical modeling. Among many modeling approaches, spectral graph model (SGM) is distinctive as it has a closed-form solution of the wide-band frequency spectra of brain oscillations, requiring only global biophysically interpretable parameters. While SGM is parsimonious in parameters, the determination of SGM parameters is non-trivial. Prior works on SGM determine the parameters through a computational intensive annealing algorithm, which only provides a point estimate with no confidence intervals for parameter estimates. To fill this gap, we incorporate the simulation-based inference (SBI) algorithm and develop a Bayesian procedure for inferring the posterior distribution of the SGM parameters. Furthermore, using SBI dramatically reduces the computational burden for inferring the SGM parameters. We evaluate the proposed SBI-SGM framework on the resting-state magnetoencephalography recordings from healthy subjects and show that the proposed procedure has similar performance to the annealing algorithm in recovering power spectra and the spatial distribution of the alpha frequency band. In addition, we also analyze the correlations among the parameters and their uncertainty with the posterior distribution which can not be done with annealing inference. These analyses provide a richer understanding of the interactions among biophysical parameters of the SGM. In general, the use of simulation-based Bayesian inference enables robust and efficient computations of generative model parameter uncertainties and may pave the way for the use of generative models in clinical translation applications.
]]></description>
<dc:creator>Jin, H.</dc:creator>
<dc:creator>Verma, P.</dc:creator>
<dc:creator>Jiang, F.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:date>2023-03-02</dc:date>
<dc:identifier>doi:10.1101/2023.03.01.530704</dc:identifier>
<dc:title><![CDATA[Bayesian Inference of a Spectral Graph Model for Brain Oscillations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.27.530254v1?rss=1">
<title>
<![CDATA[
Structure-based discovery of cannabinoid-1 receptor agonists with reduced side effects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.27.530254v1?rss=1</link>
<description><![CDATA[
Large library docking can reveal unexpected chemotypes that complement the structures of biological targets. Seeking new agonists for the cannabinoid-1 receptor (CB1R), we docked 74 million tangible molecules, prioritizing 46 high ranking ones for de novo synthesis and testing. Nine were active by radioligand competition, a 20% hit-rate. Structure-based optimization of one of the most potent of these (Ki = 0.7 {micro}M) led to  4042, a 1.9 nM ligand and a full CB1R agonist. A cryo-EM structure of the purified enantiomer of  4042 ( 1350) in complex with CB1R-Gi1 confirmed its docked pose. The new agonist was strongly analgesic, with generally a 5-10-fold therapeutic window over sedation and catalepsy and no observable conditioned place preference. These findings suggest that new cannabinoid chemotypes may disentangle characteristic cannabinoid side-effects from their analgesia, supporting the further development of cannabinoids as pain therapeutics.
]]></description>
<dc:creator>Tummino, T. A.</dc:creator>
<dc:creator>Iliopoulos-Tsoutsouvas, C.</dc:creator>
<dc:creator>Braz, J. M.</dc:creator>
<dc:creator>O'Brien, E. S.</dc:creator>
<dc:creator>Stein, R. M.</dc:creator>
<dc:creator>Craik, V.</dc:creator>
<dc:creator>Tran, N. K.</dc:creator>
<dc:creator>Ganapathy, S.</dc:creator>
<dc:creator>Shiimura, Y.</dc:creator>
<dc:creator>Tong, F.</dc:creator>
<dc:creator>Ho, T. C.</dc:creator>
<dc:creator>Radchenko, D. S.</dc:creator>
<dc:creator>Moroz, Y. S.</dc:creator>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>Rosado, S. R.</dc:creator>
<dc:creator>Bhardwaj, K.</dc:creator>
<dc:creator>Benitez, J.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Kandasamy, H.</dc:creator>
<dc:creator>Normand, C.</dc:creator>
<dc:creator>Semache, M.</dc:creator>
<dc:creator>Sabbagh, L.</dc:creator>
<dc:creator>Glenn, I.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Kumar, K. K.</dc:creator>
<dc:creator>Makriyannis, A.</dc:creator>
<dc:creator>Basbaum, A. I.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2023-03-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.27.530254</dc:identifier>
<dc:title><![CDATA[Structure-based discovery of cannabinoid-1 receptor agonists with reduced side effects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.28.530401v1?rss=1">
<title>
<![CDATA[
Multivariate prediction of cognitive performance from the sleep electroencephalogram 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.28.530401v1?rss=1</link>
<description><![CDATA[
Human cognitive performance is a key function whose biological foundations have been partially revealed by genetic and brain imaging studies. The sleep electroencephalogram (EEG) is tightly linked to structural and functional features of the central nervous system and serves as another promising biomarker. We used data from MrOS, a large cohort of older men and cross- validated regularized regression to link sleep EEG features to cognitive performance in cross- sectional analyses. In independent validation samples 2.5-10% of variance in cognitive performance can be accounted for by sleep EEG features, depending on the covariates used. Demographic characteristics accounted for more covariance between sleep EEG and cognition than health variables, and consequently reduced this association by a greater degree, but even with the strictest covariate sets a statistically significant association was present. Sigma power in NREM and beta power in REM sleep were associated with better cognitive performance, while theta power in REM sleep was associated with worse performance, with no substantial effect of coherence and other sleep EEG metrics. Our findings show that cognitive performance is associated with the sleep EEG (r=0.283), with the strongest effect ascribed to spindle- frequency activity. This association becomes weaker after adjusting for demographic (r=0.186) and health variables (r=0.155), but its resilience to covariate inclusion suggest that it also partially reflects trait-like differences in cognitive ability.
]]></description>
<dc:creator>Ujma, P. P.</dc:creator>
<dc:creator>Bodizs, R.</dc:creator>
<dc:creator>Dresler, M.</dc:creator>
<dc:creator>Simor, P.</dc:creator>
<dc:creator>Purcell, S.</dc:creator>
<dc:creator>Stone, K.</dc:creator>
<dc:creator>Yaffe, K.</dc:creator>
<dc:creator>Redline, S.</dc:creator>
<dc:date>2023-03-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.28.530401</dc:identifier>
<dc:title><![CDATA[Multivariate prediction of cognitive performance from the sleep electroencephalogram]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.05.531055v1?rss=1">
<title>
<![CDATA[
Zfp106 binds to G-quadruplex RNAs and inhibits RAN translation and formation of RNA foci caused by G4C2 repeats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.05.531055v1?rss=1</link>
<description><![CDATA[
Expansion of intronic GGGGCC repeats in the C9orf72 gene causes amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Transcription of the expanded repeats results in the formation of RNA-containing nuclear foci and altered RNA metabolism. In addition, repeat-associated non-AUG (RAN) translation of the expanded GGGGCC-repeat sequence results in the production of highly toxic dipeptide-repeat (DPR) proteins. GGGGCC-repeat-containing transcripts form G-quadruplexes, which are associated with formation of RNA foci and RAN translation. Zfp106, an RNA-binding protein essential for motor neuron survival in mice, suppresses neurotoxicity in a Drosophila model of C9orf72 ALS via a previously unknown mechanism. Here, we show that Zfp106 inhibits formation of RNA foci and significantly reduces RAN translation caused by GGGGCC-repeats in mammalian cells. Further, we show that Zfp106 binds to RNA G-quadruplexes and causes a conformational change in the G-quadruplex structure formed by GGGGCC-repeats. These data suggest that Zfp106 suppresses GGGGCC repeat-mediated cytotoxicity through alteration of the repeats G-quadruplex structure.
]]></description>
<dc:creator>Celona, B.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Dang, B.</dc:creator>
<dc:creator>Kratochvil, H. T.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:creator>Black, B. L.</dc:creator>
<dc:date>2023-03-05</dc:date>
<dc:identifier>doi:10.1101/2023.03.05.531055</dc:identifier>
<dc:title><![CDATA[Zfp106 binds to G-quadruplex RNAs and inhibits RAN translation and formation of RNA foci caused by G4C2 repeats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.03.531032v1?rss=1">
<title>
<![CDATA[
PI3K block restores age-dependent neurovascular coupling defects associated with cerebral small vessel disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.03.531032v1?rss=1</link>
<description><![CDATA[
Neurovascular coupling (NVC), a vital physiological process that rapidly and precisely directs localized blood flow to the most active regions of the brain, is accomplished in part by the vast network of cerebral capillaries acting as a sensory web capable of detecting increases in neuronal activity and orchestrating the dilation of upstream parenchymal arterioles. Here, we report a Col4a1 mutant mouse model of cerebral small vessel disease (cSVD) with age-dependent defects in capillary-to-arteriole dilation, functional hyperemia in the brain, and memory. The fundamental defect in aged mutant animals was the depletion of the minor membrane phospholipid phosphatidylinositol 4,5 bisphosphate (PIP2) in brain capillary endothelial cells, leading to the loss of inwardly rectifier K+ (Kir2.1) channel activity. Blocking phosphatidylinositol-3-kinase (PI3K), an enzyme that diminishes the bioavailability of PIP2 by converting it to phosphatidylinositol (3,4,5)-trisphosphate (PIP3), restored Kir2.1 channel activity, capillary-to-arteriole dilation, and functional hyperemia. In longitudinal studies, chronic PI3K inhibition also improved the memory function of aged Col4a1 mutant mice. Our data suggest that PI3K inhibition is a viable therapeutic strategy for treating defective NVC and cognitive impairment associated with cSVD.

One-sentence summaryPI3K inhibition rescues neurovascular coupling defects in cerebral small vessel disease.
]]></description>
<dc:creator>Thakore, P.</dc:creator>
<dc:creator>Yamasaki, E.</dc:creator>
<dc:creator>Ali, S.</dc:creator>
<dc:creator>Sanchez Solano, A.</dc:creator>
<dc:creator>Labelle-Dumais, C.</dc:creator>
<dc:creator>Gao, X.</dc:creator>
<dc:creator>Chaumeil, M. M.</dc:creator>
<dc:creator>Gould, D. B.</dc:creator>
<dc:creator>Earley, S.</dc:creator>
<dc:date>2023-03-06</dc:date>
<dc:identifier>doi:10.1101/2023.03.03.531032</dc:identifier>
<dc:title><![CDATA[PI3K block restores age-dependent neurovascular coupling defects associated with cerebral small vessel disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.05.531189v1?rss=1">
<title>
<![CDATA[
Massively parallel characterization of transcriptional regulatory elements in three diverse human cell types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.05.531189v1?rss=1</link>
<description><![CDATA[
The human genome contains millions of candidate cis-regulatory elements (CREs) with cell-type-specific activities that shape both health and myriad disease states. However, we lack a functional understanding of the sequence features that control the activity and cell-type-specific features of these CREs. Here, we used lentivirus-based massively parallel reporter assays (lentiMPRAs) to test the regulatory activity of over 680,000 sequences, representing a nearly comprehensive set of all annotated CREs among three cell types (HepG2, K562, and WTC11), finding 41.7% to be functional. By testing sequences in both orientations, we find promoters to have significant strand orientation effects. We also observe that their 200 nucleotide cores function as non-cell-type-specific  on switches providing similar expression levels to their associated gene. In contrast, enhancers have weaker orientation effects, but increased tissue-specific characteristics. Utilizing our lentiMPRA data, we develop sequence-based models to predict CRE function with high accuracy and delineate regulatory motifs. Testing an additional lentiMPRA library encompassing 60,000 CREs in all three cell types, we further identified factors that determine cell-type specificity. Collectively, our work provides an exhaustive catalog of functional CREs in three widely used cell lines, and showcases how large-scale functional measurements can be used to dissect regulatory grammar.
]]></description>
<dc:creator>Agarwal, V.</dc:creator>
<dc:creator>Inoue, F.</dc:creator>
<dc:creator>Schubach, M.</dc:creator>
<dc:creator>Martin, B.</dc:creator>
<dc:creator>Dash, P.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Sohota, A.</dc:creator>
<dc:creator>Noble, W.</dc:creator>
<dc:creator>Yardimci, G.</dc:creator>
<dc:creator>Kircher, M.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2023-03-06</dc:date>
<dc:identifier>doi:10.1101/2023.03.05.531189</dc:identifier>
<dc:title><![CDATA[Massively parallel characterization of transcriptional regulatory elements in three diverse human cell types]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.05.531224v1?rss=1">
<title>
<![CDATA[
KSHV uses viral IL6 to expand infected immunosuppressive macrophages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.05.531224v1?rss=1</link>
<description><![CDATA[
Kaposis sarcoma-associated herpesvirus (KSHV) is an oncogenic double-stranded DNA virus and the etiologic agent of Kaposis sarcoma and hyperinflammatory lymphoproliferative disorders. Understanding the mechanism by which KSHV increases the infected cell population is crucial for curing KSHV-associated diseases. Here we demonstrate that KSHV preferentially infects CD14+ monocytes and sustains viral replication through the viral interleukin-6 (vIL6)-mediated activation of STAT1 and 3. Using vIL6-sufficient and vIL6-deficient recombinant KSHV, we demonstrated that vIL6 plays a critical role in promoting the proliferation and differentiation of KSHV-infected monocytes into macrophages. The macrophages derived from vIL6-sufficient KSHV infection showed a distinct transcriptional profile of elevated IFN-pathway activation with immune suppression and were compromised in T-cell stimulation function compared to those from vIL6-deficient KSHV infection or uninfected control. These results highlight a clever strategy, in which KSHV utilizes vIL6 to secure its viral pool by expanding infected dysfunctional macrophages. This mechanism also facilitates KSHV to escape from host immune surveillance and to establish a lifelong infection. 160

SummaryKSHV causes multiple inflammatory diseases, however, the underlying mechanism is not clear. Shimoda et al. demonstrate that KSHV preferentially infects monocytes and utilizes virally encoded interleukin-6 to expand and deregulate infected monocytes. This helps the virus escape from host immune surveillance.
]]></description>
<dc:creator>Shimoda, M.</dc:creator>
<dc:creator>Inagaki, T.</dc:creator>
<dc:creator>Davis, R. R.</dc:creator>
<dc:creator>Merleev, A.</dc:creator>
<dc:creator>Tepper, C.</dc:creator>
<dc:creator>Maverakis, E. M.</dc:creator>
<dc:creator>Izumiya, Y.</dc:creator>
<dc:date>2023-03-06</dc:date>
<dc:identifier>doi:10.1101/2023.03.05.531224</dc:identifier>
<dc:title><![CDATA[KSHV uses viral IL6 to expand infected immunosuppressive macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.06.531356v1?rss=1">
<title>
<![CDATA[
Transcriptional adaptation of drug-tolerant Mycobacterium tuberculosis in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.06.531356v1?rss=1</link>
<description><![CDATA[
Transcriptome evaluation of Mycobacterium tuberculosis in the lungs of laboratory animals during long-term treatment has been limited by extremely low abundance of bacterial mRNA relative to eukaryotic RNA. Here we report a targeted amplification RNA sequencing method called SEARCH-TB. After confirming that SEARCH-TB recapitulates conventional RNA-seq in vitro, we applied SEARCH-TB to Mycobacterium tuberculosis-infected BALB/c mice treated for up to 28 days with the global standard isoniazid, rifampin, pyrazinamide, and ethambutol regimen. We compared results in mice with 8-day exposure to the same regimen in vitro. After treatment of mice for 28 days, SEARCH-TB suggested broad suppression of genes associated with bacterial growth, transcription, translation, synthesis of rRNA proteins and immunogenic secretory peptides. Adaptation of drug-stressed Mycobacterium tuberculosis appeared to include a metabolic transition from ATP-maximizing respiration towards lower-efficiency pathways, modification and recycling of cell wall components, large-scale regulatory reprogramming, and reconfiguration of efflux pumps expression. Despite markedly different expression at pre-treatment baseline, murine and in vitro samples had broadly similar transcriptional change during treatment. The differences observed likely indicate the importance of immunity and pharmacokinetics in the mouse. By elucidating the long-term effect of tuberculosis treatment on bacterial cellular processes in vivo, SEARCH-TB represents a highly granular pharmacodynamic monitoring tool with potential to enhance evaluation of new regimens and thereby accelerate progress towards a new generation of more effective tuberculosis treatment.
]]></description>
<dc:creator>Wynn, E. A.</dc:creator>
<dc:creator>Dide-Agossou, C.</dc:creator>
<dc:creator>Reichlen, M.</dc:creator>
<dc:creator>Rossmassler, K.</dc:creator>
<dc:creator>Al Mubarak, R.</dc:creator>
<dc:creator>Reid, J. J.</dc:creator>
<dc:creator>Tabor, S. T.</dc:creator>
<dc:creator>Born, S. E.</dc:creator>
<dc:creator>Ransom, M. R.</dc:creator>
<dc:creator>Davidson, R. M.</dc:creator>
<dc:creator>Walton, K. N.</dc:creator>
<dc:creator>Benoit, J. B.</dc:creator>
<dc:creator>Bauman, A. A.</dc:creator>
<dc:creator>Massoudi, L. M.</dc:creator>
<dc:creator>Dolganov, G.</dc:creator>
<dc:creator>Nahid, P.</dc:creator>
<dc:creator>Voskuil, M. I.</dc:creator>
<dc:creator>Robertson, G. T.</dc:creator>
<dc:creator>Moore, C. M.</dc:creator>
<dc:creator>Walter, N. D.</dc:creator>
<dc:date>2023-03-06</dc:date>
<dc:identifier>doi:10.1101/2023.03.06.531356</dc:identifier>
<dc:title><![CDATA[Transcriptional adaptation of drug-tolerant Mycobacterium tuberculosis in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.06.531220v1?rss=1">
<title>
<![CDATA[
Generation of Schwann cell derived melanocytes from hPSCs identifies pro-metastatic factors in melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.06.531220v1?rss=1</link>
<description><![CDATA[
Summary/AbstractThe neural crest (NC) is highly multipotent and generates diverse lineages in the developing embryo. However, spatiotemporally distinct NC populations display differences in fate potential, such as increased gliogenic and parasympathetic potential from later migrating, nerve-associated Schwann cell precursors (SCPs). Interestingly, while melanogenic potential is shared by both early migrating NC and SCPs, differences in melanocyte identity resulting from differentiation through these temporally distinct progenitors have not been determined. Here, we leverage a human pluripotent stem cell (hPSC) model of NC temporal patterning to comprehensively characterize human NC heterogeneity, fate bias, and lineage development. We captured the transition of NC differentiation between temporally and transcriptionally distinct melanogenic progenitors and identified modules of candidate transcription factor and signaling activity associated with this transition. For the first time, we established a protocol for the directed differentiation of melanocytes from hPSCs through a SCP intermediate, termed trajectory 2 (T2) melanocytes. Leveraging an existing protocol for differentiating early NC-derived melanocytes, termed trajectory 1 (T1), we performed the first comprehensive comparison of transcriptional and functional differences between these distinct melanocyte populations, revealing differences in pigmentation and unique expression of transcription factors, ligands, receptors and surface markers. We found a significant link between the T2 melanocyte transcriptional signature and decreased survival in melanoma patients in the cancer genome atlas (TCGA). We performed an in vivo CRISPRi screen of T1 and T2 melanocyte signature genes in a human melanoma cell line and discovered several T2-specific markers that promote lung metastasis in mice. We further demonstrated that one of these factors, SNRPB, regulates the splicing of transcripts involved in metastasis relevant functions such as migration, cell adhesion and proliferation. Overall, this study identifies distinct developmental trajectories as a source of diversity in melanocytes and implicates the unique molecular signature of SCP-derived melanocytes in metastatic melanoma.
]]></description>
<dc:creator>Samuel, R. M.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Maynard, A.</dc:creator>
<dc:creator>Gaylord, E. A.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Bhat, S.</dc:creator>
<dc:creator>Majd, H.</dc:creator>
<dc:creator>Richter, M. N.</dc:creator>
<dc:creator>Elder, N.</dc:creator>
<dc:creator>Le, D.</dc:creator>
<dc:creator>Nguyen, P.</dc:creator>
<dc:creator>Shibata, B.</dc:creator>
<dc:creator>Losa Llabata, M.</dc:creator>
<dc:creator>Selleri, L.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:creator>Darmanis, S.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:date>2023-03-07</dc:date>
<dc:identifier>doi:10.1101/2023.03.06.531220</dc:identifier>
<dc:title><![CDATA[Generation of Schwann cell derived melanocytes from hPSCs identifies pro-metastatic factors in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.06.531355v1?rss=1">
<title>
<![CDATA[
DNAJB8 oligomerization is mediated by an aromatic-rich motif that is dispensable for substrate activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.06.531355v1?rss=1</link>
<description><![CDATA[
J-domain protein (JDP) molecular chaperones have emerged as central players that maintain a healthy proteome. The diverse members of the JDP family function as monomers/dimers and a small subset assemble into micron-sized oligomers. The oligomeric JDP members have eluded structural characterization due to their low-complexity, intrinsically disordered middle domains. This in turn, obscures the biological significance of these larger oligomers in protein folding processes. Here, we identified a short, aromatic motif within DNAJB8, that drives self-assembly through {nu}-{nu} stacking and determined its X-ray structure. We show that mutations in the motif disrupt DNAJB8 oligomerization in vitro and in cells. DNAJB8 variants that are unable to assemble bind to misfolded tau seeds more specifically and retain capacity to reduce protein aggregation in vitro and in cells. We propose a new model for DNAJB8 function in which the sequences in the low-complexity domains play distinct roles in assembly and substrate activity.

HIGHLIGHTSDNAJB8 oligomerization is mediated by a short phenylalanine-based motif in the S/T domain

Mutation of a single phenylalanine yields a monomeric form of DNAJB8

Monomeric DNABJ8 binds to an aggregation-prone substrate

Monomeric DNAJB8 retains substrate aggregation prevention activity
]]></description>
<dc:creator>Ryder, B. D.</dc:creator>
<dc:creator>Boyer, D. R.</dc:creator>
<dc:creator>Ustyantseva, E.</dc:creator>
<dc:creator>Mendoza-Oliva, A.</dc:creator>
<dc:creator>Kuska, M.</dc:creator>
<dc:creator>Wydorski, P. M.</dc:creator>
<dc:creator>Sawaya, M.</dc:creator>
<dc:creator>Diamond, M. I.</dc:creator>
<dc:creator>Eisenberg, D.</dc:creator>
<dc:creator>Kampinga, H. H.</dc:creator>
<dc:creator>Joachimiak, L. A.</dc:creator>
<dc:date>2023-03-07</dc:date>
<dc:identifier>doi:10.1101/2023.03.06.531355</dc:identifier>
<dc:title><![CDATA[DNAJB8 oligomerization is mediated by an aromatic-rich motif that is dispensable for substrate activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.09.531574v1?rss=1">
<title>
<![CDATA[
Evolutionary constraint and innovation across hundreds of placental mammals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.09.531574v1?rss=1</link>
<description><![CDATA[
Evolutionary constraint and acceleration are powerful, cell-type agnostic measures of functional importance. Previous studies in mammals were limited by species number and reliance on human-referenced alignments. We explore the evolution of placental mammals, including humans, through reference-free whole-genome alignment of 240 species and protein-coding alignments for 428 species. We estimate 10.7% of the human genome is evolutionarily constrained. We resolve constraint to single nucleotides, pinpointing functional positions, and refine and expand by over seven-fold the catalog of ultraconserved elements. Overall, 48.5% of constrained bases are as yet unannotated, suggesting yet-to-be-discovered functional importance. Using species-level phenotypes and an updated phylogeny, we associate coding and regulatory variation with olfaction and hibernation. Focusing on biodiversity conservation, we identify genomic metrics that predict species at risk of extinction.
]]></description>
<dc:creator>Christmas, M. J.</dc:creator>
<dc:creator>Kaplow, I. M.</dc:creator>
<dc:creator>Genereux, D. P.</dc:creator>
<dc:creator>Dong, M. X.</dc:creator>
<dc:creator>Hughes, G. M.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Sullivan, P. F.</dc:creator>
<dc:creator>Hindle, A. G.</dc:creator>
<dc:creator>Andrews, G.</dc:creator>
<dc:creator>Armstrong, J. C.</dc:creator>
<dc:creator>Bianchi, M.</dc:creator>
<dc:creator>Breit, A. M.</dc:creator>
<dc:creator>Diekhans, M.</dc:creator>
<dc:creator>Fanter, C.</dc:creator>
<dc:creator>Foley, N. M.</dc:creator>
<dc:creator>Goodman, D. B.</dc:creator>
<dc:creator>Goodman, L.</dc:creator>
<dc:creator>Keough, K. C.</dc:creator>
<dc:creator>Kirilenko, B.</dc:creator>
<dc:creator>Kowalczyk, A.</dc:creator>
<dc:creator>Lawless, C.</dc:creator>
<dc:creator>Lind, A. L.</dc:creator>
<dc:creator>Meadows, J. R. S.</dc:creator>
<dc:creator>Moreira, L. R.</dc:creator>
<dc:creator>Redlich, R. W.</dc:creator>
<dc:creator>Ryan, L.</dc:creator>
<dc:creator>Swofford, R.</dc:creator>
<dc:creator>Valenzuela, A.</dc:creator>
<dc:creator>Wagner, F.</dc:creator>
<dc:creator>Wallerman, O.</dc:creator>
<dc:creator>Brown, A. R.</dc:creator>
<dc:creator>Damas, J.</dc:creator>
<dc:creator>Fan, K.</dc:creator>
<dc:creator>Gatesy, J.</dc:creator>
<dc:creator>Grimshaw, J.</dc:creator>
<dc:creator>Johnson, J.</dc:creator>
<dc:creator>Kozyrev, S. V.</dc:creator>
<dc:creator>Lawler, A. J.</dc:creator>
<dc:creator>Marinescu, V. D.</dc:creator>
<dc:creator>Morrill, K. M.</dc:creator>
<dc:creator>Osmanski, A.</dc:creator>
<dc:creator>Paulat, N. S.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2023-03-09</dc:date>
<dc:identifier>doi:10.1101/2023.03.09.531574</dc:identifier>
<dc:title><![CDATA[Evolutionary constraint and innovation across hundreds of placental mammals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.09.531820v1?rss=1">
<title>
<![CDATA[
Aberrant phase separation is a common killing strategy of positively charged peptides in biology and human disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.09.531820v1?rss=1</link>
<description><![CDATA[
Positively charged repeat peptides are emerging as key players in neurodegenerative diseases. These peptides can perturb diverse cellular pathways but a unifying framework for how such promiscuous toxicity arises has remained elusive. We used mass-spectrometry-based proteomics to define the protein targets of these neurotoxic peptides and found that they all share similar sequence features that drive their aberrant condensation with these positively charged peptides. We trained a machine learning algorithm to detect such sequence features and unexpectedly discovered that this mode of toxicity is not limited to human repeat expansion disorders but has evolved countless times across the tree of life in the form of cationic antimicrobial and venom peptides. We demonstrate that an excess in positive charge is necessary and sufficient for this killer activity, which we name  polycation poisoning. These findings reveal an ancient and conserved mechanism and inform ways to leverage its design rules for new generations of bioactive peptides.
]]></description>
<dc:creator>Boeynaems, S.</dc:creator>
<dc:creator>Ma, X. R.</dc:creator>
<dc:creator>Yeong, V.</dc:creator>
<dc:creator>Ginell, G. M.</dc:creator>
<dc:creator>Chen, J.-H.</dc:creator>
<dc:creator>Blum, J. A.</dc:creator>
<dc:creator>Nakayama, L.</dc:creator>
<dc:creator>Sanyal, A.</dc:creator>
<dc:creator>Briner, A.</dc:creator>
<dc:creator>Van Haver, D.</dc:creator>
<dc:creator>Pauwels, J.</dc:creator>
<dc:creator>Ekman, A.</dc:creator>
<dc:creator>Schmidt, H. B.</dc:creator>
<dc:creator>Sundararajan, K.</dc:creator>
<dc:creator>Porta, L.</dc:creator>
<dc:creator>Lasker, K.</dc:creator>
<dc:creator>Larabell, C.</dc:creator>
<dc:creator>Hayashi, M. A. F.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:creator>Impens, F.</dc:creator>
<dc:creator>Obermeyer, A.</dc:creator>
<dc:creator>Holehouse, A. S.</dc:creator>
<dc:creator>Gitler, A. D.</dc:creator>
<dc:date>2023-03-09</dc:date>
<dc:identifier>doi:10.1101/2023.03.09.531820</dc:identifier>
<dc:title><![CDATA[Aberrant phase separation is a common killing strategy of positively charged peptides in biology and human disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.07.531597v1?rss=1">
<title>
<![CDATA[
Torch-eCpG: A fast and scalable eQTM mapper for thousands of molecular phenotypes with graphical processing units 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.07.531597v1?rss=1</link>
<description><![CDATA[
BackgroundGene expression may be regulated by the DNA methylation of regulatory elements in cis, distal, and trans regions. One method to evaluate the relationship between DNA methylation and gene expression is the mapping of expression quantitative trait methylation (eQTM) loci (also called expression associated CpG loci, eCpG). However, no open-source tools are available to provide eQTM mapping. In addition, eQTM mapping can involve a large number of comparisons which may prevent the analyses due to limitations of computational resources. Here, we describe Torch-eCpG, an open-source tool to perform eQTM mapping that includes an optimized implementation that can use the graphical processing unit (GPU) to reduce runtime.

ResultsWe demonstrate the analyses using the tool are reproducible, up to 18x faster using the GPU, and scale linearly with increasing methylation loci.

ConclusionsTorch-eCpG is a fast, reliable, and scalable tool to perform eQTM mapping. Source code for Torch-eCpG is available at https://github.com/kordk/torch-ecpg.
]]></description>
<dc:creator>Kober, K. M.</dc:creator>
<dc:creator>Berger, L. G.</dc:creator>
<dc:creator>Roy, R.</dc:creator>
<dc:creator>Olshen, A.</dc:creator>
<dc:date>2023-03-10</dc:date>
<dc:identifier>doi:10.1101/2023.03.07.531597</dc:identifier>
<dc:title><![CDATA[Torch-eCpG: A fast and scalable eQTM mapper for thousands of molecular phenotypes with graphical processing units]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.08.531806v1?rss=1">
<title>
<![CDATA[
Inhibition of muscarinic receptor signaling protects human enteric inhibitory neurons against platin chemotherapy toxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.08.531806v1?rss=1</link>
<description><![CDATA[
GI toxicity is a common dose-limiting adverse effect of platin chemotherapy treatment. Up to 50% of cancer survivors continue to experience symptoms of chronic constipation or diarrhea induced by their chemotherapy for many years after their treatment. This drug toxicity is largely attributed to damage to enteric neurons that innervate the GI tract and control GI motility. The mechanisms responsible for platin-induced enteric neurotoxicity and potential preventative strategies have remained unknown. Here, we use human pluripotent stem cell derived enteric neurons to establish a new model system capable of uncovering the mechanism of platin-induced enteric neuropathy. Utilizing this scalable system, we performed a high throughput screen and identified drug candidates and pathways involved in the disease. Our analyses revealed that excitotoxicity through muscarinic cholinergic signaling is a key driver of platin-induced enteric neuropathy. Using single nuclei transcriptomics and functional assays, we discovered that this disease mechanism leads to increased susceptibility of specific neuronal subtypes, including inhibitory nitrergic neurons, to platins. Histological assessment of the enteric nervous system in platin-treated patients confirmed the selective loss of nitrergic neurons. Finally, we demonstrated that pharmacological and genetic inhibition of muscarinic cholinergic signaling is sufficient to rescue enteric neurons from platin excitotoxicity in vitro and can prevent platin-induced constipation and degeneration of nitrergic neurons in mice. These studies define the mechanisms of platin-induced enteric neuropathy and serve as a framework for uncovering cell type-specific manifestations of cellular stress underlying numerous intractable peripheral neuropathies.
]]></description>
<dc:creator>Richter, M. N.</dc:creator>
<dc:creator>Farahvashi, S.</dc:creator>
<dc:creator>Samuel, R. M.</dc:creator>
<dc:creator>Majd, H.</dc:creator>
<dc:creator>Chemel, A. K.</dc:creator>
<dc:creator>Ramirez, J. T.</dc:creator>
<dc:creator>Majd, A.</dc:creator>
<dc:creator>Scantlen, M. D.</dc:creator>
<dc:creator>Elder, N.</dc:creator>
<dc:creator>Cesiulis, A.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Joshi, T.</dc:creator>
<dc:creator>Keefe, M. G.</dc:creator>
<dc:creator>Samiakalantari, B.</dc:creator>
<dc:creator>Turkalj, E. M.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Arab, A.</dc:creator>
<dc:creator>Yin, K.</dc:creator>
<dc:creator>Culbertson, B.</dc:creator>
<dc:creator>Vora, B.</dc:creator>
<dc:creator>Xiong, C.</dc:creator>
<dc:creator>Kattah, M. G.</dc:creator>
<dc:creator>Irannejad, R.</dc:creator>
<dc:creator>Kroetz, D. L.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:date>2023-03-10</dc:date>
<dc:identifier>doi:10.1101/2023.03.08.531806</dc:identifier>
<dc:title><![CDATA[Inhibition of muscarinic receptor signaling protects human enteric inhibitory neurons against platin chemotherapy toxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.10.531987v1?rss=1">
<title>
<![CDATA[
Title: Leveraging Base Pair Mammalian Constraint to Understand Genetic Variation and Human Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.10.531987v1?rss=1</link>
<description><![CDATA[
Although thousands of genomic regions have been associated with heritable human diseases, attempts to elucidate biological mechanisms are impeded by a general inability to discern which genomic positions are functionally important. Evolutionary constraint is a powerful predictor of function that is agnostic to cell type or disease mechanism. Here, single base phyloP scores from the whole genome alignment of 240 placental mammals identified 3.5% of the human genome as significantly constrained, and likely functional. We compared these scores to large-scale genome annotation, genome-wide association studies (GWAS), copy number variation, clinical genetics findings, and cancer data sets. Evolutionarily constrained positions are enriched for variants explaining common disease heritability (more than any other functional annotation). Our results improve variant annotation but also highlight that the regulatory landscape of the human genome still needs to be further explored and linked to disease.
]]></description>
<dc:creator>Sullivan, P. F.</dc:creator>
<dc:creator>Meadows, J. R. S.</dc:creator>
<dc:creator>Gazal, S.</dc:creator>
<dc:creator>Phan, B. N.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Genereux, D. P.</dc:creator>
<dc:creator>Dong, M. X.</dc:creator>
<dc:creator>Bianchi, M.</dc:creator>
<dc:creator>Andrews, G.</dc:creator>
<dc:creator>Sakthikumar, S.</dc:creator>
<dc:creator>Nordin, J.</dc:creator>
<dc:creator>Roy, A.</dc:creator>
<dc:creator>Christmas, M. J.</dc:creator>
<dc:creator>Marinescu, V. D.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Wallerman, O.</dc:creator>
<dc:creator>Xue, J. R.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Yao, S.</dc:creator>
<dc:creator>Sun, Q.</dc:creator>
<dc:creator>Szatkiewicz, J.</dc:creator>
<dc:creator>Wen, J.</dc:creator>
<dc:creator>Huckins, L. M.</dc:creator>
<dc:creator>Lawler, A. J.</dc:creator>
<dc:creator>Keough, K. C.</dc:creator>
<dc:creator>Zheng, Z.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:creator>Wray, N. R.</dc:creator>
<dc:creator>Johnson, J.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Zoonomia Consortium,</dc:creator>
<dc:creator>Paten, B.</dc:creator>
<dc:creator>Reilly, S. K.</dc:creator>
<dc:creator>Hughes, G. M.</dc:creator>
<dc:creator>Weng, Z.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Pfenning, A. R.</dc:creator>
<dc:creator>Forsberg-Nilsson, K.</dc:creator>
<dc:creator>Karlsson, E. K.</dc:creator>
<dc:creator>Lindblad-Toh, K.</dc:creator>
<dc:date>2023-03-10</dc:date>
<dc:identifier>doi:10.1101/2023.03.10.531987</dc:identifier>
<dc:title><![CDATA[Title: Leveraging Base Pair Mammalian Constraint to Understand Genetic Variation and Human Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.08.531782v1?rss=1">
<title>
<![CDATA[
Cyclone: an accessible pipeline to analyze, evaluate and optimize multiparametric cytometry data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.08.531782v1?rss=1</link>
<description><![CDATA[
In the past decade, high-dimensional single cell technologies have revolutionized basic and translational immunology research and are now a key element of the toolbox used by scientists to study the immune system. However, analysis of the data generated by these approaches often requires clustering algorithms and dimensionality reduction representation which are computationally intense and difficult to evaluate and optimize. Here we present Cyclone, an analysis pipeline integrating dimensionality reduction, clustering, evaluation and optimization of clustering resolution, and downstream visualization tools facilitating the analysis of a wide range of cytometry data. We benchmarked and validated Cyclone on mass cytometry (CyTOF), full spectrum fluorescence-based cytometry, and multiplexed immunofluorescence (IF) in a variety of biological contexts, including infectious diseases and cancer. In each instance, Cyclone not only recapitulates gold standard immune cell identification, but also enables the unsupervised identification of lymphocytes and mononuclear phagocytes subsets that are associated with distinct biological features. Altogether, the Cyclone pipeline is a versatile and accessible pipeline for performing, optimizing, and evaluating clustering on variety of cytometry datasets which will further power immunology research and provide a scaffold for biological discovery.
]]></description>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Jaszczak, R. G.</dc:creator>
<dc:creator>Kwok, I.</dc:creator>
<dc:creator>Carey, N. D.</dc:creator>
<dc:creator>Courau, T.</dc:creator>
<dc:creator>Bunis, D.</dc:creator>
<dc:creator>Samad, B.</dc:creator>
<dc:creator>Avanesyan, L.</dc:creator>
<dc:creator>Chew, N. W.</dc:creator>
<dc:creator>Stenske, S.</dc:creator>
<dc:creator>Jespersen, J. M.</dc:creator>
<dc:creator>Publicover, J.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Naser, M.</dc:creator>
<dc:creator>Rao, A. A.</dc:creator>
<dc:creator>Lupin-Jimenez, L.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Cooper, S.</dc:creator>
<dc:creator>Baron, J.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:date>2023-03-11</dc:date>
<dc:identifier>doi:10.1101/2023.03.08.531782</dc:identifier>
<dc:title><![CDATA[Cyclone: an accessible pipeline to analyze, evaluate and optimize multiparametric cytometry data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.10.531984v1?rss=1">
<title>
<![CDATA[
Unsupervised pattern discovery in spatial gene expression atlas reveals mouse brain regions beyond established ontology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.10.531984v1?rss=1</link>
<description><![CDATA[
The rapid growth of large-scale spatial gene expression data demands efficient and reliable computational tools to extract major trends of gene expression in their native spatial context. Here, we used stability-driven unsupervised learning (i.e. staNMF) to identify principal patterns (PPs) of 3D gene expression profiles and understand spatial gene distribution and anatomical localization at the whole mouse brain level. Our subsequent spatial correlation analysis systematically compared the PPs to known anatomical regions and ontology from the Allen Mouse Brain Atlas using spatial neighborhoods. We demonstrate that our stable and spatially coherent PPs, whose linear combinations accurately approximate the spatial gene data, are highly correlated with combinations of expert-annotated brain regions. These PPs yield a new brain ontology based purely on spatial gene expression. Our PP identification approach outperforms principal component analysis (PCA) and typical clustering algorithms on the same task. Moreover, we show that the stable PPs reveal marked regional imbalance of brainwide genetic architecture, leading to region-specific marker genes and gene co-expression networks. Our findings highlight the advantages of stability-driven machine learning for plausible biological discovery from dense spatial gene expression data that are infeasible by conventional manual approaches.
]]></description>
<dc:creator>Cahill, R.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Lee, A.</dc:creator>
<dc:creator>Zeng, H.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Tasic, B.</dc:creator>
<dc:creator>Abbasi-Asl, R.</dc:creator>
<dc:date>2023-03-12</dc:date>
<dc:identifier>doi:10.1101/2023.03.10.531984</dc:identifier>
<dc:title><![CDATA[Unsupervised pattern discovery in spatial gene expression atlas reveals mouse brain regions beyond established ontology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.11.532161v1?rss=1">
<title>
<![CDATA[
A systematic search for RNA structural switches across the human transcriptome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.11.532161v1?rss=1</link>
<description><![CDATA[
RNA structural switches are key regulators of gene expression in bacteria, yet their characterization in Metazoa remains limited. Here we present SwitchSeeker, a comprehensive computational and experimental approach for systematic identification of functional RNA structural switches. We applied SwitchSeeker to the human transcriptome and identified 245 putative RNA switches. To validate our approach, we characterized a previously unknown RNA switch in the 3UTR of the RORC transcript. In vivo DMS-MaPseq, coupled with cryogenic electron microscopy, confirmed its existence as two alternative structural conformations. Furthermore, we used genome-scale CRISPR screens to identify trans factors that regulate gene expression through this RNA structural switch. We found that nonsense-mediated mRNA decay acts on this element in a conformation-specific manner. SwitchSeeker provides an unbiased, experimentally-driven method for discovering RNA structural switches that shape the eukaryotic gene expression landscape.
]]></description>
<dc:creator>Khoroshkin, M.</dc:creator>
<dc:creator>Asarnow, D.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Winters, A.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Zhou, S. K.</dc:creator>
<dc:creator>Zhou, S.</dc:creator>
<dc:creator>Palka, C.</dc:creator>
<dc:creator>Fish, L.</dc:creator>
<dc:creator>Ansel, K. M.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2023-03-12</dc:date>
<dc:identifier>doi:10.1101/2023.03.11.532161</dc:identifier>
<dc:title><![CDATA[A systematic search for RNA structural switches across the human transcriptome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.11.532186v1?rss=1">
<title>
<![CDATA[
Pharmacologic Activation of an Integrated Stress Response Kinase Promotes Mitochondrial Remodeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.11.532186v1?rss=1</link>
<description><![CDATA[
The integrated stress response (ISR) comprises the eIF2 kinases PERK, GCN2, HRI, and PKR, which induce translational and transcriptional signaling in response to diverse insults. Deficiencies in PERK signaling lead to mitochondrial dysfunction and contribute to the pathogenesis of numerous diseases. We define the potential for pharmacologic activation of compensatory eIF2 kinases to rescue ISR signaling and promote mitochondrial adaptation in PERK-deficient cells. We show that the HRI activator BtdCPU and GCN2 activator halofuginone promote ISR signaling and rescue ER stress sensitivity in PERK-deficient cells. However, BtdCPU induces mitochondrial depolarization, leading to mitochondrial fragmentation and activation of the OMA1-DELE1-HRI signaling axis. In contrast, halofuginone promotes mitochondrial elongation and adaptive mitochondrial respiration, mimicking regulation induced by PERK. This shows halofuginone can compensate for deficiencies in PERK signaling and promote adaptive mitochondrial remodeling, highlighting the potential for pharmacologic ISR activation to mitigate mitochondrial dysfunction and motivating the pursuit of highly-selective ISR activators.
]]></description>
<dc:creator>Perea, V.</dc:creator>
<dc:creator>Baron, K. R.</dc:creator>
<dc:creator>Dolina, V.</dc:creator>
<dc:creator>Aviles, G.</dc:creator>
<dc:creator>Rosarda, J. D.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Wiseman, L.</dc:creator>
<dc:date>2023-03-12</dc:date>
<dc:identifier>doi:10.1101/2023.03.11.532186</dc:identifier>
<dc:title><![CDATA[Pharmacologic Activation of an Integrated Stress Response Kinase Promotes Mitochondrial Remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.12.531757v1?rss=1">
<title>
<![CDATA[
A molecular glue approach to control the half-life of CRISPR-based technologies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.12.531757v1?rss=1</link>
<description><![CDATA[
Cas9 is a programmable nuclease that has furnished transformative technologies, including base editors and transcription modulators (e.g., CRISPRi/a), but several applications of these technologies, including therapeutics, mandatorily require precision control of their half-life. For example, such control can help avert any potential immunological and adverse events in clinical trials. Current genome editing technologies to control the half-life of Cas9 are slow, have lower activity, involve fusion of large response elements (> 230 amino acids), utilize expensive controllers with poor pharmacological attributes, and cannot be implemented in vivo on several CRISPR-based technologies. We report a general platform for half-life control using the molecular glue, pomalidomide, that binds to a ubiquitin ligase complex and a response-element bearing CRISPR-based technology, thereby causing the latters rapid ubiquitination and degradation. Using pomalidomide, we were able to control the half-life of large CRISPR-based technologies (e.g., base editors, CRISPRi) and small anti-CRISPRs that inhibit such technologies, allowing us to build the first examples of on-switch for base editors. The ability to switch on, fine-tune and switch-off CRISPR-based technologies with pomalidomide allowed complete control over their activity, specificity, and genome editing outcome. Importantly, the miniature size of the response element and favorable pharmacological attributes of the drug pomalidomide allowed control of activity of base editor in vivo using AAV as the delivery vehicle. These studies provide methods and reagents to precisely control the dosage and half-life of CRISPR-based technologies, propelling their therapeutic development.
]]></description>
<dc:creator>Sreekanth, V.</dc:creator>
<dc:creator>Jan, M.</dc:creator>
<dc:creator>Zhao, K. T.</dc:creator>
<dc:creator>Lim, D.</dc:creator>
<dc:creator>Davis, J. R.</dc:creator>
<dc:creator>McConkey, M.</dc:creator>
<dc:creator>Kovalcik, V.</dc:creator>
<dc:creator>Barkal, S.</dc:creator>
<dc:creator>Law, B. K.</dc:creator>
<dc:creator>Fife, J.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Vinyard, M. E.</dc:creator>
<dc:creator>Becerra, B.</dc:creator>
<dc:creator>Kampmann, M. E.</dc:creator>
<dc:creator>Sherwood, R. I.</dc:creator>
<dc:creator>Pinello, L.</dc:creator>
<dc:creator>Liu, D. R.</dc:creator>
<dc:creator>Ebert, B. L.</dc:creator>
<dc:creator>Choudhary, A.</dc:creator>
<dc:date>2023-03-12</dc:date>
<dc:identifier>doi:10.1101/2023.03.12.531757</dc:identifier>
<dc:title><![CDATA[A molecular glue approach to control the half-life of CRISPR-based technologies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.13.532469v1?rss=1">
<title>
<![CDATA[
Aversive experience drives offline ensemble reactivation to link memories across days 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.13.532469v1?rss=1</link>
<description><![CDATA[
Memories are encoded in neural ensembles during learning and stabilized by post-learning reactivation. Integrating recent experiences into existing memories ensures that memories contain the most recently available information, but how the brain accomplishes this critical process remains unknown. Here we show that in mice, a strong aversive experience drives the offline ensemble reactivation of not only the recent aversive memory but also a neutral memory formed two days prior, linking the fear from the recent aversive memory to the previous neutral memory. We find that fear specifically links retrospectively, but not prospectively, to neutral memories across days. Consistent with prior studies, we find reactivation of the recent aversive memory ensemble during the offline period following learning. However, a strong aversive experience also increases co-reactivation of the aversive and neutral memory ensembles during the offline period. Finally, the expression of fear in the neutral context is associated with reactivation of the shared ensemble between the aversive and neutral memories. Taken together, these results demonstrate that strong aversive experience can drive retrospective memory-linking through the offline co-reactivation of recent memory ensembles with memory ensembles formed days prior, providing a neural mechanism by which memories can be integrated across days.
]]></description>
<dc:creator>Zaki, Y.</dc:creator>
<dc:creator>Pennington, Z. T.</dc:creator>
<dc:creator>Morales-Rodriguez, D.</dc:creator>
<dc:creator>Francisco, T. R.</dc:creator>
<dc:creator>LaBanca, A. R.</dc:creator>
<dc:creator>Dong, Z.</dc:creator>
<dc:creator>Segura, S. C.</dc:creator>
<dc:creator>Silva, A. J.</dc:creator>
<dc:creator>Shuman, T.</dc:creator>
<dc:creator>Fenton, A.</dc:creator>
<dc:creator>Rajan, K.</dc:creator>
<dc:creator>Cai, D. J.</dc:creator>
<dc:date>2023-03-14</dc:date>
<dc:identifier>doi:10.1101/2023.03.13.532469</dc:identifier>
<dc:title><![CDATA[Aversive experience drives offline ensemble reactivation to link memories across days]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.14.532632v1?rss=1">
<title>
<![CDATA[
Comparative analysis between RNA-seq and single-molecule RNA FISH indicates that the pyrimidine nucleobase idoxuridine (IdU) globally amplifies transcriptional noise 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.14.532632v1?rss=1</link>
<description><![CDATA[
Stochastic fluctuations (noise) in transcription generate substantial cell-to-cell variability, but the physiological roles of noise have remained difficult to determine in the absence of generalized noise-modulation approaches. Previous single-cell RNA-sequencing (scRNA-seq) suggested that the pyrimidine-base analog (5-iodo-2-deoxyuridine, IdU) could generally amplify noise without substantially altering mean-expression levels but scRNA-seq technical drawbacks potentially obscured the penetrance of IdU-induced transcriptional noise amplification. Here we quantify global-vs.-partial penetrance of IdU-induced noise amplification by assessing scRNA-seq data using numerous normalization algorithms and directly quantifying noise using single-molecule RNA FISH (smFISH) for a panel of genes from across the transcriptome. Alternate scRNA-seq analyses indicate IdU-induced noise amplification for ~90% of genes, and smFISH data verified noise amplification for ~90% of tested genes. Collectively, this analysis indicates which scRNA-seq algorithms are appropriate for quantifying noise and argues that IdU is a globally penetrant noise-enhancer molecule that could enable investigations of the physiological impacts of transcriptional noise.

Graphical Abstract O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY
]]></description>
<dc:creator>Calia, G. P.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Zuckerman, B.</dc:creator>
<dc:creator>Weinberger, L.</dc:creator>
<dc:date>2023-03-15</dc:date>
<dc:identifier>doi:10.1101/2023.03.14.532632</dc:identifier>
<dc:title><![CDATA[Comparative analysis between RNA-seq and single-molecule RNA FISH indicates that the pyrimidine nucleobase idoxuridine (IdU) globally amplifies transcriptional noise]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.15.532602v1?rss=1">
<title>
<![CDATA[
Hox genes modulate physical forces to differentially shape small and large intestinal epithelia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.15.532602v1?rss=1</link>
<description><![CDATA[
The small and large intestines arise from a common primordial gut tube but ultimately become specialized in both form and function. While the midgut forms villi, the hindgut develops flat, brain-like sulci that resolve into heterogeneous outgrowths. Gut compartment identities are demarcated early in development via Hox genes, which are highly conserved, master regulators of spatial patterning in the embryo. Yet, how these factors trigger regional morphogenesis has remained a mystery. Combining mechanical measurements and mathematical modeling, we demonstrate that the posterior Hox gene Hoxd13 regulates biophysical phenomena that shape the hindgut lumen. We further show that Hoxd13 acts through the TGF{beta} pathway to thicken, stiffen, and promote isotropic growth of the subepithelial mesenchyme; together, these features generate hindgut surface patterns. TGF{beta}, in turn, promotes collagen deposition to affect mesenchymal geometry and growth. We thus identify a cascade of events downstream of genetic identity that direct posterior intestinal morphogenesis.
]]></description>
<dc:creator>Gill, H. K.</dc:creator>
<dc:creator>Yin, S.</dc:creator>
<dc:creator>Nerurkar, N. L.</dc:creator>
<dc:creator>Lawlor, J. C.</dc:creator>
<dc:creator>Huycke, T. R.</dc:creator>
<dc:creator>Mahadevan, L.</dc:creator>
<dc:creator>Tabin, C. J.</dc:creator>
<dc:date>2023-03-15</dc:date>
<dc:identifier>doi:10.1101/2023.03.15.532602</dc:identifier>
<dc:title><![CDATA[Hox genes modulate physical forces to differentially shape small and large intestinal epithelia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.15.532818v1?rss=1">
<title>
<![CDATA[
A protein hydroxylase couples epithelial membrane biology to nucleolar ribosome biogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.15.532818v1?rss=1</link>
<description><![CDATA[
Jumonji-C (JmjC) ribosomal protein hydroxylases are an ancient class of oxygen- and Fe(II)-dependent oxygenases that spawned the wider JmjC family and Histone Lysine Demethylases (KDMs) in eukaryotes. Myc-induced Antigen (MINA) has been implicated in ribosome biogenesis and was assigned as a nucleolar-localized JmjC histidyl hydroxylase of the large ribosomal subunit protein RPL27A, consistent with reports that it supports cell growth and viability in a variety of tumor cell types. Reported roles in diverse aspects of disease biology may be consistent with additional MINA functions, although the molecular mechanisms involved remain unclear. Here, we describe an extra-nucleolar interaction of MINA with the Hinge domain of the membrane-associated guanylate kinase, MPP6. We show that MINA promotes the expression and membrane localization of MPP6 and that the MINA-MPP6 pathway is required for epithelial tight junction integrity and barrier function. The function of MINA in this novel pathway is suppressed by ribosomal RNA transcription and the nucleolar MINA interactome. In this way, MINA couples epithelial membrane biology to nucleolar ribosome biogenesis. Our work sheds light on how quiescent cells lose adhesion as they switch to proliferative states associated with increased ribosome biogenesis.
]]></description>
<dc:creator>Hendrix, E.</dc:creator>
<dc:creator>Andrijes, R.</dc:creator>
<dc:creator>Boora, U.</dc:creator>
<dc:creator>Kaur, A.</dc:creator>
<dc:creator>Bundred, J. R.</dc:creator>
<dc:creator>Zayer, A.</dc:creator>
<dc:creator>Heilig, R.</dc:creator>
<dc:creator>Westrip, C. A. E.</dc:creator>
<dc:creator>Fletcher, S. F.</dc:creator>
<dc:creator>Eaton, C. D.</dc:creator>
<dc:creator>Kennedy, T. J.</dc:creator>
<dc:creator>Piasecka, S.</dc:creator>
<dc:creator>Fischer, R.</dc:creator>
<dc:creator>Smerdon, S. J.</dc:creator>
<dc:creator>Coleman, M. L.</dc:creator>
<dc:date>2023-03-16</dc:date>
<dc:identifier>doi:10.1101/2023.03.15.532818</dc:identifier>
<dc:title><![CDATA[A protein hydroxylase couples epithelial membrane biology to nucleolar ribosome biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.15.532845v1?rss=1">
<title>
<![CDATA[
HBV rewires liver cancer signaling by altering PP2A complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.15.532845v1?rss=1</link>
<description><![CDATA[
Infection by hepatitis B virus (HBV) increases risk for liver cancer by inducing inflammation, cellular stress and cell death. To elucidate the molecular pathways by which HBV promotes cancer development and progression, we used affinity purification mass spectrometry to comprehensively map a network of 145 physical interactions between HBV and human host proteins in hepatocellular carcinoma (HCC). We find that viral proteins target host factors that are preferentially mutated in non-HBV-associated HCC, implicating cancer pathways whose interaction with HBV plays a role in HCC. Focusing on proteins that directly interact with the HBV oncoprotein X (HBx), we show that HBx remodels the PP2A phosphatase complex, altering its effect on tumor signaling. HBx excludes striatin-family regulatory subunits from PP2A, causing Hippo kinase activation and unmasking a requirement for mTOR complex 2 to maintain expression of the YAP oncoprotein in HCC. Thus, HBV rewires HCC to expose potentially targetable signaling dependencies.

SignificancePrecision medicine has revolutionized cancer treatment but remains elusive for HCC. We used proteomics to define HBV/host interactions and integrated them with HCC mutations. The results implicate modifiers of HCC behavior via remodeling of host complexes and illuminate new biological mechanisms in advanced disease for therapeutic investigation.
]]></description>
<dc:creator>Pitea, A.</dc:creator>
<dc:creator>Turnham, R.</dc:creator>
<dc:creator>Eckhardt, M.</dc:creator>
<dc:creator>Jang, G. M.</dc:creator>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Lim, H. C.</dc:creator>
<dc:creator>Choi, A.</dc:creator>
<dc:creator>Von Dollen, J.</dc:creator>
<dc:creator>Levin, R. S.</dc:creator>
<dc:creator>Webber, J. T.</dc:creator>
<dc:creator>McCarthy, E.</dc:creator>
<dc:creator>Hu, J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Che, L.</dc:creator>
<dc:creator>Chan, G.</dc:creator>
<dc:creator>Kelley, R. K.</dc:creator>
<dc:creator>Swaney, D.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Bandyopadhyay, S.</dc:creator>
<dc:creator>Theis, F. J.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:creator>Ideker, T.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Gordan, J.</dc:creator>
<dc:date>2023-03-16</dc:date>
<dc:identifier>doi:10.1101/2023.03.15.532845</dc:identifier>
<dc:title><![CDATA[HBV rewires liver cancer signaling by altering PP2A complexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.16.532850v1?rss=1">
<title>
<![CDATA[
Group 2 innate lymphoid cells promote inhibitory synapse development and social behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.16.532850v1?rss=1</link>
<description><![CDATA[
The innate immune system plays essential roles in brain synaptic development, and immune dysregulation is implicated in neurodevelopmental diseases. Here we show that a subset of innate lymphocytes (group 2 innate lymphoid cells, ILC2s) is required for cortical inhibitory synapse maturation and adult social behavior. ILC2s expanded in the developing meninges and produced a surge of their canonical cytokine Interleukin-13 (IL-13) between postnatal days 5-15. Loss of ILC2s decreased cortical inhibitory synapse numbers in the postnatal period where as ILC2 transplant was sufficient to increase inhibitory synapse numbers. Deletion of the IL-4/IL-13 receptor (Il4ra) from inhibitory neurons phenocopied the reduction inhibitory synapses. Both ILC2 deficient and neuronal Il4ra deficient animals had similar and selective impairments in adult social behavior. These data define a type 2 immune circuit in early life that shapes adult brain function.

One sentence summaryType 2 innate lymphoid cells and Interleukin-13 promote inhibitory synapse development.
]]></description>
<dc:creator>Barron, J. J.</dc:creator>
<dc:creator>Mroz, N. M.</dc:creator>
<dc:creator>Taloma, S. E.</dc:creator>
<dc:creator>Dahlgren, M. W.</dc:creator>
<dc:creator>Ortiz-Carpena, J. F.</dc:creator>
<dc:creator>Dorman, L. C.</dc:creator>
<dc:creator>Vainchtein, I. D.</dc:creator>
<dc:creator>Escoubas, C. C.</dc:creator>
<dc:creator>Molofsky, A. B.</dc:creator>
<dc:creator>Molofsky, A. V.</dc:creator>
<dc:date>2023-03-17</dc:date>
<dc:identifier>doi:10.1101/2023.03.16.532850</dc:identifier>
<dc:title><![CDATA[Group 2 innate lymphoid cells promote inhibitory synapse development and social behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.17.533028v1?rss=1">
<title>
<![CDATA[
Sensory cortical ensembles exhibit differential coupling to ripples in distinct hippocampal subregions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.17.533028v1?rss=1</link>
<description><![CDATA[
Cortical neurons activated during recent experiences often reactivate with dorsal hippocampal CA1 sharp-wave ripples (SWRs) during subsequent rest. Less is known about cortical interactions with intermediate hippocampal CA1, whose connectivity, functions, and SWRs differ from those of dorsal CA1. We identified three clusters of visual cortical excitatory neurons that are excited together with either dorsal or intermediate CA1 SWRs, or suppressed before both SWRs. Neurons in each cluster were distributed across primary and higher visual cortices and co-active even in the absence of SWRs. These ensembles exhibited similar visual responses but different coupling to thalamus and pupil-indexed arousal. We observed a consistent activity sequence: (i) suppression of SWR-suppressed cortical neurons, (ii) thalamic silence, and (iii) activation of the cortical ensemble preceding and predicting intermediate CA1 SWRs. We propose that the coordinated dynamics of these ensembles relay visual experiences to distinct hippocampal subregions for incorporation into different cognitive maps.
]]></description>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>K Namboodiri, V. M.</dc:creator>
<dc:creator>Jung, M. W.</dc:creator>
<dc:creator>Andermann, M. L.</dc:creator>
<dc:date>2023-03-18</dc:date>
<dc:identifier>doi:10.1101/2023.03.17.533028</dc:identifier>
<dc:title><![CDATA[Sensory cortical ensembles exhibit differential coupling to ripples in distinct hippocampal subregions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.17.533206v1?rss=1">
<title>
<![CDATA[
Cryo-EM structure of the human Sirtuin 6-nucleosome complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.17.533206v1?rss=1</link>
<description><![CDATA[
Sirtuin 6 (SIRT6) is a multifaceted protein deacetylase/deacylase and a major target for small-molecule modulators of longevity and cancer. In the context of chromatin, SIRT6 removes acetyl groups from histone H3 in nucleosomes, but the molecular basis for its nucleosomal substrate preference is unknown. Our cryo-electron microscopy structure of human SIRT6 in complex with the nucleosome shows that the catalytic domain of SIRT6 pries DNA from the nucleosomal entry-exit site and exposes the histone H3 N-terminal helix, while the SIRT6 zinc-binding domain binds to the histone acidic patch using an arginine anchor. In addition, SIRT6 forms an inhibitory interaction with the C-terminal tail of histone H2A. The structure provides insights into how SIRT6 can deacetylate both H3 K9 and H3 K56.

TeaserThe structure of the SIRT6 deacetylase/nucleosome complex suggests how the enzyme acts on both histone H3 K9 and K56 residues.
]]></description>
<dc:creator>Chio, U. S.</dc:creator>
<dc:creator>Rechiche, O.</dc:creator>
<dc:creator>Bryll, A. R.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Feldman, J.</dc:creator>
<dc:creator>Peterson, C.</dc:creator>
<dc:creator>Tan, S.</dc:creator>
<dc:creator>Armache, J.-P.</dc:creator>
<dc:date>2023-03-18</dc:date>
<dc:identifier>doi:10.1101/2023.03.17.533206</dc:identifier>
<dc:title><![CDATA[Cryo-EM structure of the human Sirtuin 6-nucleosome complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.17.533220v1?rss=1">
<title>
<![CDATA[
Catalytic and non-catalytic mechanisms of histone H4 lysine 20 methyltransferase SUV420H1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.17.533220v1?rss=1</link>
<description><![CDATA[
The intricate regulation of chromatin plays a key role in controlling genome architecture and accessibility. Histone lysine methyltransferases regulate chromatin by catalyzing the methylation of specific histone residues but are also hypothesized to have equally important non-catalytic roles. SUV420H1 di- and tri-methylates histone H4 lysine 20 (H4K20me2/me3) and plays crucial roles in DNA replication, repair, and heterochromatin formation, and is dysregulated in several cancers. Many of these processes were linked to its catalytic activity. However, deletion and inhibition of SUV420H1 have shown distinct phenotypes suggesting the enzyme likely has uncharacterized non-catalytic activities. To characterize the catalytic and non-catalytic mechanisms SUV420H1 uses to modify chromatin, we determined cryo- EM structures of SUV420H1 complexes with nucleosomes containing histone H2A or its variant H2A.Z. Our structural, biochemical, biophysical, and cellular analyses reveal how both SUV420H1 recognizes its substrate and H2A.Z stimulates its activity, and show that SUV420H1 binding to nucleosomes causes a dramatic detachment of nucleosomal DNA from histone octamer. We hypothesize that this detachment increases DNA accessibility to large macromolecular complexes, a prerequisite for DNA replication and repair. We also show that SUV420H1 can promote chromatin condensates, another non-catalytic role that we speculate is needed for its heterochromatin functions. Together, our studies uncover and characterize the catalytic and non-catalytic mechanisms of SUV420H1, a key histone methyltransferase that plays an essential role in genomic stability.
]]></description>
<dc:creator>Abini-Agbomson, S.</dc:creator>
<dc:creator>Gretarsson, K.</dc:creator>
<dc:creator>Shih, R. M.</dc:creator>
<dc:creator>Hsieh, L.</dc:creator>
<dc:creator>Lou, T.</dc:creator>
<dc:creator>De Ioannes, P.</dc:creator>
<dc:creator>Vasilyev, N.</dc:creator>
<dc:creator>Lee, R.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Simon, M.</dc:creator>
<dc:creator>Armache, J.-P.</dc:creator>
<dc:creator>Nudler, E.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Lu, C.</dc:creator>
<dc:creator>Armache, K.-J.</dc:creator>
<dc:date>2023-03-18</dc:date>
<dc:identifier>doi:10.1101/2023.03.17.533220</dc:identifier>
<dc:title><![CDATA[Catalytic and non-catalytic mechanisms of histone H4 lysine 20 methyltransferase SUV420H1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.19.533346v1?rss=1">
<title>
<![CDATA[
Comparative landscape of genetic dependencies in human and chimpanzee stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.19.533346v1?rss=1</link>
<description><![CDATA[
Comparative studies of great apes provide a window into our evolutionary past, but the extent and identity of cellular differences that emerged during hominin evolution remain largely unexplored. We established a comparative loss-of-function approach to evaluate whether changes in human cells alter requirements for essential genes. By performing genome-wide CRISPR interference screens in human and chimpanzee pluripotent stem cells, we identified 75 genes with species-specific effects on cellular proliferation. These genes comprised coherent processes, including cell cycle progression and lysosomal signaling, which we determined to be human-derived by comparison with orangutan cells. Human-specific robustness to CDK2 and CCNE1 depletion persisted in neural progenitor cells, providing support for the G1-phase length hypothesis as a potential evolutionary mechanism in human brain expansion. Our findings demonstrate that evolutionary changes in human cells can reshape the landscape of essential genes and establish a platform for systematically uncovering latent cellular and molecular differences between species.
]]></description>
<dc:creator>She, R.</dc:creator>
<dc:creator>Fair, T.</dc:creator>
<dc:creator>Schaefer, N. K.</dc:creator>
<dc:creator>Saunders, R. A.</dc:creator>
<dc:creator>Pavlovic, B. J.</dc:creator>
<dc:creator>Weissman, J. S.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:date>2023-03-20</dc:date>
<dc:identifier>doi:10.1101/2023.03.19.533346</dc:identifier>
<dc:title><![CDATA[Comparative landscape of genetic dependencies in human and chimpanzee stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.17.533227v1?rss=1">
<title>
<![CDATA[
A Spatiotemporal Map of Co-Receptor Signaling Networks Underlying B Cell Activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.17.533227v1?rss=1</link>
<description><![CDATA[
The B cell receptor (BCR) signals together with a multi-component co-receptor complex to initiate B cell activation in response to antigen binding. This process underlies nearly every aspect of proper B cell function. Here, we take advantage of peroxidase-catalyzed proximity labeling combined with quantitative mass spectrometry to track B cell co-receptor signaling dynamics from 10 seconds to 2 hours after BCR stimulation. This approach enables tracking of 2,814 proximity-labeled proteins and 1,394 quantified phosphosites and provides an unbiased and quantitative molecular map of proteins recruited to the vicinity of CD19, the key signaling subunit of the co-receptor complex. We detail the recruitment kinetics of essential signaling effectors to CD19 following activation, and then identify new mediators of B cell activation. In particular, we show that the glutamate transporter SLC1A1 is responsible for mediating rapid metabolic reprogramming immediately downstream of BCR stimulation and for maintaining redox homeostasis during B cell activation. This study provides a comprehensive map of the BCR signaling pathway and a rich resource for uncovering the complex signaling networks that regulate B cell activation.
]]></description>
<dc:creator>Susa, K. J.</dc:creator>
<dc:creator>Bradshaw, G. A.</dc:creator>
<dc:creator>Eisert, R. J.</dc:creator>
<dc:creator>Schillling, C. M.</dc:creator>
<dc:creator>Kalocsay, M.</dc:creator>
<dc:creator>Blacklow, S. C.</dc:creator>
<dc:creator>Kruse, A. C.</dc:creator>
<dc:date>2023-03-21</dc:date>
<dc:identifier>doi:10.1101/2023.03.17.533227</dc:identifier>
<dc:title><![CDATA[A Spatiotemporal Map of Co-Receptor Signaling Networks Underlying B Cell Activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.22.533709v1?rss=1">
<title>
<![CDATA[
Mitigation of chromosome loss in clinical CRISPR-Cas9-engineered T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.22.533709v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas9 genome editing has enabled advanced T cell therapies, but occasional loss of the targeted chromosome remains a safety concern. To investigate whether Cas9-induced chromosome loss is a universal phenomenon and evaluate its clinical significance, we conducted a systematic analysis in primary human T cells. Arrayed and pooled CRISPR screens revealed that chromosome loss was generalizable across the genome and resulted in partial and entire loss of the chromosome, including in pre-clinical chimeric antigen receptor T cells. T cells with chromosome loss persisted for weeks in culture, implying the potential to interfere with clinical use. A modified cell manufacturing process, employed in our first-in-human clinical trial of Cas9-engineered T cells,1 dramatically reduced chromosome loss while largely preserving genome editing efficacy. Expression of p53 correlated with protection from chromosome loss observed in this protocol, suggesting both a mechanism and strategy for T cell engineering that mitigates this genotoxicity in the clinic.
]]></description>
<dc:creator>Tsuchida, C. A.</dc:creator>
<dc:creator>Brandes, N.</dc:creator>
<dc:creator>Bueno, R.</dc:creator>
<dc:creator>Trinidad, M.</dc:creator>
<dc:creator>Mazumder, T.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Hwang, B.</dc:creator>
<dc:creator>Chang, C.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Hopkins, C. R.</dc:creator>
<dc:creator>Parker, K. R.</dc:creator>
<dc:creator>Qi, Y.</dc:creator>
<dc:creator>Satpathy, A.</dc:creator>
<dc:creator>Stadtmauer, E.</dc:creator>
<dc:creator>Cate, J. H. D.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Fraietta, J. A.</dc:creator>
<dc:creator>June, C. H.</dc:creator>
<dc:creator>Chang, H. Y.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:date>2023-03-22</dc:date>
<dc:identifier>doi:10.1101/2023.03.22.533709</dc:identifier>
<dc:title><![CDATA[Mitigation of chromosome loss in clinical CRISPR-Cas9-engineered T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.19.533348v1?rss=1">
<title>
<![CDATA[
Bayesian Inference for Brain Source Imaging with Joint Estimation of Structured Low-rank Noise 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.19.533348v1?rss=1</link>
<description><![CDATA[
The inverse problem in brain source imaging is the reconstruction of brain activity from non-invasive recordings of electroencephalography (EEG) and magnetoencephalography (MEG). One key challenge is the efficient recovery of sparse brain activity when the data is corrupted by structured noise that is low-rank noise. This is often the case when there are a few active sources of environmental noise and the MEG/EEG sensor noise is highly correlated. In this paper, we propose a novel robust empirical Bayesian framework which provides us a tractable algorithm for jointly estimating a low-rank noise covariance and brain source activity. Specifically, we use a factor analysis model for the structured noise, and infer a sparse set of variance parameters for source activity, while performing Variational Bayesian inference for the noise. One key aspect of this algorithm is that it does not require any additional baseline measurements to estimate the noise covariance from the sensor data. We perform exhaustive experiments on both simulated and real datasets. Our algorithm achieves superior performance as compared to several existing benchmark algorithms.
]]></description>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Cai, C.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Hashemi, A.</dc:creator>
<dc:creator>Haufe, S.</dc:creator>
<dc:creator>Sekihara, K.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2023-03-23</dc:date>
<dc:identifier>doi:10.1101/2023.03.19.533348</dc:identifier>
<dc:title><![CDATA[Bayesian Inference for Brain Source Imaging with Joint Estimation of Structured Low-rank Noise]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.23.533424v1?rss=1">
<title>
<![CDATA[
Functional identification of language-responsive channels in individual participants in MEG investigations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.23.533424v1?rss=1</link>
<description><![CDATA[
Making meaningful inferences about the functional architecture of the language system requires the ability to refer to the same neural units across individuals and studies. Traditional brain imaging approaches align and average brains together in a common space. However, lateral frontal and temporal cortices, where the language system resides, is characterized by high structural and functional inter-individual variability, which reduces the sensitivity and functional resolution of group-averaging analyses. This issue is compounded by the fact that language areas lay in close proximity to regions of other large-scale networks with different functional profiles. A solution inspired by visual neuroscience is to identify language areas functionally in each individual brain using a  localizer task (e.g., a language comprehension task). This approach has proven productive in fMRI, yielding a number of robust and replicable findings about the language system. Here, we extend this approach to MEG. Across two experiments (one in Dutch speakers, n=19; one in English speakers, n=23), we examined neural responses to the processing of sentences and a control condition (nonword sequences). In both the time and frequency domains, we demonstrated that the topography of neural responses to language is spatially stable within individuals but varies across individuals. Consequently, analyses that take this inter-individual variability into account are characterized by greater sensitivity, compared to the group-level analyses. In summary, similar to fMRI, functional identification within individuals yields benefits in MEG, thus opening the door to future investigations of language processing including questions where whole-brain coverage and temporal resolution are both critical.
]]></description>
<dc:creator>Bruffaerts, R.</dc:creator>
<dc:creator>Pongos, A.</dc:creator>
<dc:creator>Shain, C.</dc:creator>
<dc:creator>Lipkin, B.</dc:creator>
<dc:creator>Siegelman, M.</dc:creator>
<dc:creator>Wens, V.</dc:creator>
<dc:creator>Sjogard, M.</dc:creator>
<dc:creator>Pantazis, D.</dc:creator>
<dc:creator>Blank, I.</dc:creator>
<dc:creator>Goldman, S.</dc:creator>
<dc:creator>De Tiege, X.</dc:creator>
<dc:creator>Fedorenko, E.</dc:creator>
<dc:date>2023-03-23</dc:date>
<dc:identifier>doi:10.1101/2023.03.23.533424</dc:identifier>
<dc:title><![CDATA[Functional identification of language-responsive channels in individual participants in MEG investigations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.22.533675v1?rss=1">
<title>
<![CDATA[
A Comprehensive Enumeration of the Human Proteostasis Network. 2. Components of the Autophagy-Lysosome Pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.22.533675v1?rss=1</link>
<description><![CDATA[
The condition of having a healthy, functional proteome is known as protein homeostasis, or proteostasis. Establishing and maintaining proteostasis is the province of the proteostasis network, approximately 2,700 components that regulate protein synthesis, folding, localization, and degradation. The proteostasis network is a fundamental entity in biology that is essential for cellular health and has direct relevance to many diseases of protein conformation. However, it is not well defined or annotated, which hinders its functional characterization in health and disease. In this series of manuscripts, we aim to operationally define the human proteostasis network by providing a comprehensive, annotated list of its components. We provided in a previous manuscript a list of chaperones and folding enzymes as well as the components that make up the machineries for protein synthesis, protein trafficking into and out of organelles, and organelle-specific degradation pathways. Here, we provide a curated list of 838 unique high-confidence components of the autophagy-lysosome pathway, one of the two major protein degradation systems in human cells.
]]></description>
<dc:creator>The Proteostasis Consortium,</dc:creator>
<dc:creator>Elsasser, S.</dc:creator>
<dc:creator>Elia, L. P.</dc:creator>
<dc:creator>Morimoto, R. I.</dc:creator>
<dc:creator>Powers, E. T.</dc:creator>
<dc:creator>Finley, D.</dc:creator>
<dc:creator>Costa, B.</dc:creator>
<dc:creator>Budron, M.</dc:creator>
<dc:creator>Tokuno, Z.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Iyer, R. G.</dc:creator>
<dc:creator>Barth, B.</dc:creator>
<dc:creator>Mockler, E.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Richardson, R. A. K.</dc:creator>
<dc:creator>Stoeger, T.</dc:creator>
<dc:creator>Tan, E. P.</dc:creator>
<dc:creator>Xiao, Q.</dc:creator>
<dc:creator>Cole, C. M.</dc:creator>
<dc:creator>Massey, L. A.</dc:creator>
<dc:creator>Garza, D.</dc:creator>
<dc:creator>Kelly, J. W.</dc:creator>
<dc:creator>Rainbolt, T. K.</dc:creator>
<dc:creator>Chou, C.-C.</dc:creator>
<dc:creator>Masto, V. B.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:creator>Nixon, R. A.</dc:creator>
<dc:date>2023-03-24</dc:date>
<dc:identifier>doi:10.1101/2023.03.22.533675</dc:identifier>
<dc:title><![CDATA[A Comprehensive Enumeration of the Human Proteostasis Network. 2. Components of the Autophagy-Lysosome Pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.22.533843v1?rss=1">
<title>
<![CDATA[
DIP-MS: A novel ultra-deep interaction proteomics for the deconvolution of protein complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.22.533843v1?rss=1</link>
<description><![CDATA[
Most, if not all, proteins are organized in macromolecular assemblies, which represent key functional units regulating and catalyzing the majority of cellular processes in health and disease. Ever-advancing analytical capabilities promise to pinpoint lesions in proteome modularity driving disease phenotypes. Affinity purification of the protein of interest combined with LC-MS/MS (AP-MS) represents the method of choice to identify interacting proteins. The composition of complex isoforms concurrently present in the AP sample can however not be resolved from a single AP-MS experiment but requires computational inference from multiple time-and resource-intensive reciprocal AP-MS experiments.

In this study we introduce Deep Interactome Profiling by Mass Spectrometry (DIP-MS) which combines affinity enrichment with BN-PAGE separation, DIA mass spectrometry and deep-learning-based signal processing to resolve complex isoforms sharing the same bait protein in a single experiment.

We applied DIP-MS to probe the organisation of the human prefoldin (PFD) family of complexes, resolving distinct PFD holo- and sub-complex variants, complex-complex interactions and complex isoforms with new subunits that were experimentally validated. Our results demonstrate that DIP-MS can reveal proteome modularity at unprecedented depth and resolution and thus represents a critical steppingstone to relate a proteome state to phenotype in both healthy and diseased conditions.
]]></description>
<dc:creator>Frommelt, F.</dc:creator>
<dc:creator>Fossati, A.</dc:creator>
<dc:creator>Uliana, F.</dc:creator>
<dc:creator>Wendt, F.</dc:creator>
<dc:creator>Xue, P.</dc:creator>
<dc:creator>Heusel, M.</dc:creator>
<dc:creator>Wollscheid, B.</dc:creator>
<dc:creator>Aebersold, R.</dc:creator>
<dc:creator>Ciuffa, R.</dc:creator>
<dc:creator>Gstaiger, M.</dc:creator>
<dc:date>2023-03-24</dc:date>
<dc:identifier>doi:10.1101/2023.03.22.533843</dc:identifier>
<dc:title><![CDATA[DIP-MS: A novel ultra-deep interaction proteomics for the deconvolution of protein complexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.23.533974v1?rss=1">
<title>
<![CDATA[
Analysis of evolutionary dynamics and clonal architecture in prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.23.533974v1?rss=1</link>
<description><![CDATA[
The extent to which clinical and genomic characteristics associate with prostate cancer clonal architecture, tumor evolution, and therapeutic response remains unclear. Here, we reconstructed the clonal architecture and evolutionary trajectories of 845 prostate cancer tumors with harmonized clinical and molecular data. We observed that tumors from patients who self-reported as Black had more linear and monoclonal architectures, despite these men having higher rates of biochemical recurrence. This finding contrasts with prior observations relating polyclonal architecture to adverse clinical outcomes. Additionally, we utilized a novel approach to mutational signature analysis that leverages clonal architecture to uncover additional cases of homologous recombination and mismatch repair deficiency in primary and metastatic tumors and link the origin of mutational signatures to specific subclones. Broadly, prostate cancer clonal architecture analysis reveals novel biological insights that may be immediately clinically actionable and provide multiple opportunities for subsequent investigation.

Statement of significanceTumors from patients who self-reported as Black demonstrate linear and monoclonal evolutionary trajectories yet experience higher rates of biochemical recurrence. In addition, analysis of clonal and subclonal mutational signatures identifies additional tumors with potentially actionable alterations such as deficiencies in mismatch repair and homologous recombination.
]]></description>
<dc:creator>Conway, J.</dc:creator>
<dc:creator>Tewari, A. K.</dc:creator>
<dc:creator>Camp, S. Y.</dc:creator>
<dc:creator>Han, S.</dc:creator>
<dc:creator>Crowdis, J.</dc:creator>
<dc:creator>He, M. X.</dc:creator>
<dc:creator>Nyame, Y. A.</dc:creator>
<dc:creator>AlDubayan, S. H.</dc:creator>
<dc:creator>Schultz, N.</dc:creator>
<dc:creator>Szallasi, Z.</dc:creator>
<dc:creator>Pomerantz, M. M.</dc:creator>
<dc:creator>Freedman, M. L.</dc:creator>
<dc:creator>Fong, L.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:creator>Salari, K.</dc:creator>
<dc:creator>Van Allen, E.</dc:creator>
<dc:date>2023-03-25</dc:date>
<dc:identifier>doi:10.1101/2023.03.23.533974</dc:identifier>
<dc:title><![CDATA[Analysis of evolutionary dynamics and clonal architecture in prostate cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.24.534024v1?rss=1">
<title>
<![CDATA[
Continuous Multiplexed Phage Genome Editing Using Recombitrons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.24.534024v1?rss=1</link>
<description><![CDATA[
Bacteriophages, which naturally shape bacterial communities, can be co-opted as a biological technology to help eliminate pathogenic bacteria from our bodies and food supply1. Phage genome editing is a critical tool to engineer more effective phage technologies. However, editing phage genomes has traditionally been a low efficiency process that requires laborious screening, counter selection, or in vitro construction of modified genomes2. These requirements impose limitations on the type and throughput of phage modifications, which in turn limit our knowledge and potential for innovation. Here, we present a scalable approach for engineering phage genomes using recombitrons: modified bacterial retrons3 that generate recombineering donor DNA paired with single stranded binding and annealing proteins to integrate those donors into phage genomes. This system can efficiently create genome modifications in multiple phages without the need for counterselection. Moreover, the process is continuous, with edits accumulating in the phage genome the longer the phage is cultured with the host, and multiplexable, with different editing hosts contributing distinct mutations along the genome of a phage in a mixed culture. In lambda phage, as an example, recombitrons yield single-base substitutions at up to 99% efficiency and up to 5 distinct mutations installed on a single phage genome, all without counterselection and only a few hours of hands-on time.
]]></description>
<dc:creator>Fishman, C. B.</dc:creator>
<dc:creator>Crawford, K. D.</dc:creator>
<dc:creator>Bhattarai-Kline, S.</dc:creator>
<dc:creator>Zhang, K. A.</dc:creator>
<dc:creator>Delgado-Gonzalez, A.</dc:creator>
<dc:creator>Shipman, S. L.</dc:creator>
<dc:date>2023-03-25</dc:date>
<dc:identifier>doi:10.1101/2023.03.24.534024</dc:identifier>
<dc:title><![CDATA[Continuous Multiplexed Phage Genome Editing Using Recombitrons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.24.534105v1?rss=1">
<title>
<![CDATA[
Glioblastoma induces the recruitment and differentiation of hybrid neutrophils from skull bone marrow 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.24.534105v1?rss=1</link>
<description><![CDATA[
Tumor-associated neutrophil (TAN) effects on glioblastoma biology remain under-characterized. We show here that  hybrid neutrophils with dendritic features - including morphological complexity, expression of antigen presentation genes, and the ability to process exogenous peptide and stimulate MHCII-dependent T cell activation - accumulate intratumorally and suppress tumor growth in vivo. Trajectory analysis of patient TAN scRNA-seq identifies this phenotype as a polarization state which is distinct from canonical cytotoxic TANs and differentiates intratumorally from immature precursors absent in circulation. Rather, these hybrid-inducible immature neutrophils - which we identified in patient and murine glioblastomas - arise from local skull marrow. Through labeled skull flap transplantation and targeted ablation, we characterize calvarial marrow as a potent contributor of antitumoral myeloid APCs, including hybrid TANs and dendritic cells, which elicit T cell cytotoxicity and memory. As such, agents augmenting neutrophil egress from skull marrow - such as intracalvarial AMD3100 whose survival prolonging-effect in GBM we demonstrate - present therapeutic potential.
]]></description>
<dc:creator>Lad, M.</dc:creator>
<dc:creator>Beniwal, A.</dc:creator>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Shukla, P.</dc:creator>
<dc:creator>Jung, J.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Yagnik, G.</dc:creator>
<dc:creator>Babikir, H.</dc:creator>
<dc:creator>Nguyen, A. T.</dc:creator>
<dc:creator>Gill, S.</dc:creator>
<dc:creator>Young, J. S.</dc:creator>
<dc:creator>Lui, A.</dc:creator>
<dc:creator>Salha, D.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:date>2023-03-25</dc:date>
<dc:identifier>doi:10.1101/2023.03.24.534105</dc:identifier>
<dc:title><![CDATA[Glioblastoma induces the recruitment and differentiation of hybrid neutrophils from skull bone marrow]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.26.533356v1?rss=1">
<title>
<![CDATA[
An essential role for miR-15/16 in Treg suppression and restriction of proliferation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.26.533356v1?rss=1</link>
<description><![CDATA[
The miR-15/16 family is a highly expressed group of tumor suppressor miRNAs that target a large network of genes in T cells to restrict their cell cycle, memory formation and survival. Upon T cell activation, miR-15/16 are downregulated, allowing rapid expansion of differentiated effector T cells to mediate a sustained immune response. Here, using conditional deletion of miR-15/16 in immunosuppressive regulatory T cells (Tregs) that express FOXP3, we identify new functions of the miR-15/16 family in T cell immunity. miR-15/16 are indispensable to maintain peripheral tolerance by securing efficient suppression by a limited number of Tregs. miR-15/16-deficiency alters Treg expression of critical functional proteins including FOXP3, IL2R/CD25, CTLA4, PD-1 and IL7R/CD127, and results in accumulation of functionally impaired FOXP3loCD25loCD127hi Tregs. Excessive proliferation in the absence of miR-15/16 inhibition of cell cycle programs shifts Treg diversity and produces an effector Treg phenotype characterized by low expression of TCF1, CD25 and CD62L, and high expression of CD44. These Tregs fail to control immune activation of CD4+ effector T cells, leading to spontaneous multi-organ inflammation and increased allergic airway inflammation in a mouse model of asthma. Together, our results demonstrate that miR-15/16 expression in Tregs is essential to maintain immune tolerance.

HighlightsO_LITreg-specific miR-15/16 expression is essential to prevent systemic tissue inflammation
C_LIO_LImiR-15/16 restrict Treg proliferation and regulate expression of the key functional Treg molecules FOXP3, IL2R, CTLA4, PD-1 and IL7R
C_LIO_LImiR-15/16 limit formation of effector Tregs and is necessary for high suppressive capacity
C_LI
]]></description>
<dc:creator>Johansson, K.</dc:creator>
<dc:creator>Gagnon, J. D.</dc:creator>
<dc:creator>Zhou, S.</dc:creator>
<dc:creator>Fassett, M. S.</dc:creator>
<dc:creator>Schroeder, A. W.</dc:creator>
<dc:creator>Kageyama, R.</dc:creator>
<dc:creator>Bautista, R. A.</dc:creator>
<dc:creator>Pham, H.</dc:creator>
<dc:creator>Woodruff, P. G.</dc:creator>
<dc:creator>Ansel, K. M.</dc:creator>
<dc:date>2023-03-26</dc:date>
<dc:identifier>doi:10.1101/2023.03.26.533356</dc:identifier>
<dc:title><![CDATA[An essential role for miR-15/16 in Treg suppression and restriction of proliferation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.26.534250v1?rss=1">
<title>
<![CDATA[
SLAPSHOT reveals rapid dynamics of extracellularly exposed proteome in response to calcium-activated plasma membrane phospholipid scrambling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.26.534250v1?rss=1</link>
<description><![CDATA[
To facilitate our understanding of the often rapid and nuanced dynamics of extracellularly exposed proteomes during signaling events, it is important to devise robust workflows affording fast time resolution without biases and confounding factors. Here, we present Surface-exposed protein Labeling using PeroxidaSe, H2O2, and Tyramide-derivative (SLAPSHOT), to label extracellularly exposed proteins in a rapid, sensitive, and specific manner, while preserving cellular integrity. This experimentally simple and flexible method utilizes recombinant soluble APEX2 peroxidase that is applied to cells, thus circumventing biological perturbations, tedious engineering of tools and cells, and labeling biases. APEX2 neither requires metal cations for activity nor contains disulfide bonds, conferring versatility for a wide spectrum of experimental setups. We applied SLAPSHOT followed by quantitative mass spectrometry-based proteomics analysis to examine the immediate and extensive cell surface expansion and ensuing restorative membrane shedding upon the activation of Scott syndrome-linked TMEM16F, a ubiquitously expressed calcium-dependent phospholipid scramblase and ion channel. Time-course data ranging from one to thirty minutes of calcium stimulation using wild-type and TMEM16F deficient cells revealed intricate co-regulation of known protein families, including those in the integrin and ICAM families. Crucially, we identified proteins that are known to reside in intracellular organelles, including ER, as occupants of the freshly deposited membrane, and mitovesicles as an abundant component and contributor to the extracellularly exposed proteome. Our study not only provides the first accounts of the immediate consequences of calcium signaling on the extracellularly exposed proteome, but also presents a blueprint for the application of SLAPSHOT as a general approach for monitoring extracellularly exposed protein dynamics.

HighlightsAn enzyme-driven method to tag extracellularly exposed proteins in an unbiased manner with a superior combination of temporal resolution, spatial specificity, and sensitivity

A general approach applicable to primary and scarce cells without involving cellular engineering

Short time scale proteome dynamics of Jurkat cells with and without TMEM16F revealed by SLAPSHOT coupled with quantitative mass spectrometry provide insights into phospholipid scrambling-mediated plasma membrane remodeling
]]></description>
<dc:creator>Tuomivaara, S. T.</dc:creator>
<dc:creator>Teo, C. F.</dc:creator>
<dc:creator>Jan, Y. N.</dc:creator>
<dc:creator>Jan, L. Y.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:date>2023-03-26</dc:date>
<dc:identifier>doi:10.1101/2023.03.26.534250</dc:identifier>
<dc:title><![CDATA[SLAPSHOT reveals rapid dynamics of extracellularly exposed proteome in response to calcium-activated plasma membrane phospholipid scrambling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.27.534317v1?rss=1">
<title>
<![CDATA[
A dual, catalytic role for the fission yeast Ccr4-Not complex in gene silencing and heterochromatin spreading 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.27.534317v1?rss=1</link>
<description><![CDATA[
AbstractHeterochromatic gene silencing relies on combinatorial control by specific histone modifications, the occurrence of transcription, and/or RNA degradation. Once nucleated, heterochromatin propagates within defined chromosomal regions and is maintained throughout cell divisions to warrant proper genome expression and integrity. The fission yeast Ccr4-Not complex has been involved in gene silencing, but its relative contribution to distinct heterochromatin domains and its role in nucleation versus spreading have remained elusive. Here, we unveil major functions for Ccr4-Not in silencing and heterochromatin spreading at the mating type locus and subtelomeres. Mutations of the catalytic subunits Caf1 or Mot2, involved in RNA deadenylation and protein ubiquitinylation respectively, result in impaired propagation of H3K9me3 and massive accumulation of nucleation-distal heterochromatic transcripts. Both silencing and spreading defects are suppressed upon disruption of the heterochromatin antagonizing factor Epe1. Overall, our results position the Ccr4-Not complex as a critical, dual regulator of heterochromatic gene silencing and spreading.

Author SummaryEukaryotic genomes are partitioned into relaxed, gene-rich regions, and condensed, gene-poor domains called heterochromatin. The maintenance of heterochromatin is crucial for proper genome expression and integrity, and requires multiple factors regulating histone modifications and/or the levels of RNA molecules produced from these regions. Such effectors not only promote heterochromatin assembly but also ensure its propagation from specific nucleation sites to defined domain boundaries. However, while the mechanisms involved in initiation of heterochromatin formation have been well documented, the molecular and biochemical properties underlying its spreading remain largely elusive. By combining genetic and single-cell approaches, we report here that the fission yeast Ccr4-Not complex, a multisubunit complex conserved throughout eukaryotes, is essential for efficient heterochromatin spreading to repress expression of nucleation-distal RNAs. The two catalytic activities of the complex, RNA deadenylation and protein ubiquitinylation, are each critical, thereby defining a dual enzymatic requirement in the process.
]]></description>
<dc:creator>Challal, D.</dc:creator>
<dc:creator>Menant, A.</dc:creator>
<dc:creator>Goksal, C.</dc:creator>
<dc:creator>Leroy, E.</dc:creator>
<dc:creator>Al-Sady, B.</dc:creator>
<dc:creator>Rougemaille, M.</dc:creator>
<dc:date>2023-03-27</dc:date>
<dc:identifier>doi:10.1101/2023.03.27.534317</dc:identifier>
<dc:title><![CDATA[A dual, catalytic role for the fission yeast Ccr4-Not complex in gene silencing and heterochromatin spreading]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.28.534017v1?rss=1">
<title>
<![CDATA[
Multiplex, single-cell CRISPRa screening for cell type specific regulatory elements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.28.534017v1?rss=1</link>
<description><![CDATA[
CRISPR-based gene activation (CRISPRa) is a promising therapeutic approach for gene therapy, upregulating gene expression by targeting promoters or enhancers in a tissue/cell-type specific manner. Here, we describe an experimental framework that combines highly multiplexed perturbations with single-cell RNA sequencing (sc-RNA-seq) to identify cell-type-specific, CRISPRa-responsive cis-regulatory elements and the gene(s) they regulate. Random combinations of many gRNAs are introduced to each of many cells, which are then profiled and partitioned into test and control groups to test for effect(s) of CRISPRa perturbations of both enhancers and promoters on the expression of neighboring genes. Applying this method to a library of 493 gRNAs targeting candidate cis-regulatory elements in both K562 cells and iPSC-derived excitatory neurons, we identify gRNAs capable of specifically upregulating intended target genes and no other neighboring genes within 1 Mb, including gRNAs yielding upregulation of six autism spectrum disorder (ASD) and neurodevelopmental disorder (NDD) risk genes in neurons. A consistent pattern is that the responsiveness of individual enhancers to CRISPRa is restricted by cell type, implying a dependency on either chromatin landscape and/or additional trans-acting factors for successful gene activation. The approach outlined here may facilitate large-scale screens for gRNAs that activate therapeutically relevant genes in a cell type-specific manner.
]]></description>
<dc:creator>Chardon, F. M.</dc:creator>
<dc:creator>McDiarmid, T. A.</dc:creator>
<dc:creator>Page, N. F.</dc:creator>
<dc:creator>Martin, B. K.</dc:creator>
<dc:creator>Domcke, S.</dc:creator>
<dc:creator>Regalado, S. G.</dc:creator>
<dc:creator>Lalanne, J.-B.</dc:creator>
<dc:creator>Calderon, D.</dc:creator>
<dc:creator>Starita, L. M.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:date>2023-03-28</dc:date>
<dc:identifier>doi:10.1101/2023.03.28.534017</dc:identifier>
<dc:title><![CDATA[Multiplex, single-cell CRISPRa screening for cell type specific regulatory elements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.27.534352v1?rss=1">
<title>
<![CDATA[
Ipsilateral restriction of chromosome movement along a centrosome, and apical-basal axis during the cell cycle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.27.534352v1?rss=1</link>
<description><![CDATA[
Little is known about how distance between homologous chromosomes are controlled during the cell cycle. Here, we show that the distribution of centromere components display two discrete clusters placed to either side of the centrosome and apical/basal axis from prophase to G1 interphase. 4-Dimensional live cell imaging analysis of centromere and centrosome tracking reveals that centromeres oscillate largely within one cluster, but do not cross over to the other cluster. We propose a model of an axis-dependent ipsilateral restriction of chromosome oscillations throughout mitosis.
]]></description>
<dc:creator>Cai, P.</dc:creator>
<dc:creator>Casas, C. J.</dc:creator>
<dc:creator>Hua, L. L.</dc:creator>
<dc:creator>Mikawa, T.</dc:creator>
<dc:date>2023-03-29</dc:date>
<dc:identifier>doi:10.1101/2023.03.27.534352</dc:identifier>
<dc:title><![CDATA[Ipsilateral restriction of chromosome movement along a centrosome, and apical-basal axis during the cell cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.28.534564v1?rss=1">
<title>
<![CDATA[
Implantation of engineered adipocytes that outcompete tumors for resources suppresses cancer progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.28.534564v1?rss=1</link>
<description><![CDATA[
Tumors acquire an increased ability to obtain and metabolize nutrients. Here, we engineered and implanted adipocytes to outcompete tumors for nutrients and show that they can substantially reduce cancer progression. Growing cells or xenografts from several cancers (breast, colon, pancreas, prostate) alongside engineered human adipocytes or adipose organoids significantly suppresses cancer progression and reduces hypoxia and angiogenesis. Transplanting modulated adipocyte organoids in pancreatic or breast cancer mouse models nearby or distal from the tumor significantly suppresses its growth. To further showcase therapeutic potential, we demonstrate that co-culturing tumor organoids derived from human breast cancers with engineered patient-derived adipocytes significantly reduces cancer growth. Combined, our results introduce a novel cancer therapeutic approach, termed adipose modulation transplantation (AMT), that can be utilized for a broad range of cancers.
]]></description>
<dc:creator>Nguyen, H. P.</dc:creator>
<dc:creator>Sheng, R.</dc:creator>
<dc:creator>Murray, E.</dc:creator>
<dc:creator>Ito, Y.</dc:creator>
<dc:creator>Bruck, M.</dc:creator>
<dc:creator>Biellak, C.</dc:creator>
<dc:creator>An, K.</dc:creator>
<dc:creator>Lynce, F.</dc:creator>
<dc:creator>Dillon, D. A.</dc:creator>
<dc:creator>Magbanua, M. J. M.</dc:creator>
<dc:creator>Huppert, L. A.</dc:creator>
<dc:creator>Hammerlindl, H.</dc:creator>
<dc:creator>Esserman, L.</dc:creator>
<dc:creator>Rosenbluth, J. M.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2023-03-29</dc:date>
<dc:identifier>doi:10.1101/2023.03.28.534564</dc:identifier>
<dc:title><![CDATA[Implantation of engineered adipocytes that outcompete tumors for resources suppresses cancer progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.29.534798v1?rss=1">
<title>
<![CDATA[
Coordinated stimulation of axon regenerative and neurodegenerative transcriptional programs by Atf4 following optic nerve injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.29.534798v1?rss=1</link>
<description><![CDATA[
Stress signaling is important for determining the fates of neurons following axonal insults. Previously we showed that the stress-responsive kinase PERK contributes to injury-induced neurodegeneration (Larhammar et al., 2017). Here we show that PERK acts primarily through Activating Transcription Factor-4 (ATF4) to stimulate not only pro-apoptotic but also pro-regenerative responses following optic nerve damage. Using conditional knockout mice, we find an extensive PERK/ATF4-dependent transcriptional response that includes canonical ATF4 target genes and modest contributions by C/EBP Homologous Protein (CHOP). Overlap with c-Jun-dependent transcription suggests interplay with a parallel stress pathway that orchestrates regenerative and apoptotic responses. Accordingly, neuronal knockout of ATF4 recapitulates the neuroprotection afforded by PERK deficiency, and PERK or ATF4 knockout impairs optic axon regeneration enabled by disrupting the tumor suppressor PTEN. These findings reveal an integral role for PERK/ATF4 in coordinating neurodegenerative and regenerative responses to CNS axon injury.
]]></description>
<dc:creator>Somasundaram, P.</dc:creator>
<dc:creator>Farley, M. M.</dc:creator>
<dc:creator>Rudy, M. A.</dc:creator>
<dc:creator>Stefanoff, D. G.</dc:creator>
<dc:creator>Shah, M.</dc:creator>
<dc:creator>Goli, P.</dc:creator>
<dc:creator>Heo, J.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Tran, N. M.</dc:creator>
<dc:creator>Watkins, T. A.</dc:creator>
<dc:date>2023-03-31</dc:date>
<dc:identifier>doi:10.1101/2023.03.29.534798</dc:identifier>
<dc:title><![CDATA[Coordinated stimulation of axon regenerative and neurodegenerative transcriptional programs by Atf4 following optic nerve injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.31.535173v1?rss=1">
<title>
<![CDATA[
Few-shot learning: temporal scaling in behavioral and dopaminergic learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.31.535173v1?rss=1</link>
<description><![CDATA[
Learning the causes of rewards is necessary for survival. Thus, it is critical to understand the mechanisms of such a vital biological process. Cue-reward learning is controlled by mesolimbic dopamine and improves with spacing of cue-reward pairings. However, whether a mathematical rule governs such improvements in learning rate, and if so, whether a unifying mechanism captures this rule and dopamine dynamics during learning remain unknown. Here, we investigate the behavioral, algorithmic, and dopaminergic mechanisms governing cuereward learning rate. Across a range of conditions in mice, we show a strong, mathematically proportional relationship between both behavioral and dopaminergic learning rates and the duration between rewards. Due to this relationship, removing up to 19 out of 20 cue-reward pairings over a fixed duration has no influence on overall learning. These findings are explained by a dopamine-based model of retrospective learning, thereby providing a unified account of the biological mechanisms of learning.
]]></description>
<dc:creator>Burke, D. A.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Wu, B.</dc:creator>
<dc:creator>Lee, S. A.</dc:creator>
<dc:creator>Floeder, J. R.</dc:creator>
<dc:creator>K Namboodiri, V. M.</dc:creator>
<dc:date>2023-03-31</dc:date>
<dc:identifier>doi:10.1101/2023.03.31.535173</dc:identifier>
<dc:title><![CDATA[Few-shot learning: temporal scaling in behavioral and dopaminergic learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.30.534992v1?rss=1">
<title>
<![CDATA[
A Suite of Mouse Reagents for Studying Amelogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.30.534992v1?rss=1</link>
<description><![CDATA[
Amelogenesis, the formation of dental enamel, is driven by specialized epithelial cells called ameloblasts, which undergo successive stages of differentiation. Ameloblasts secrete enamel matrix proteins (EMPs), proteases, calcium, and phosphate ions in a stage-specific manner to form mature tooth enamel. Developmental defects in tooth enamel are common in humans, and they can greatly impact the well-being of affected individuals. Our understanding of amelogenesis and developmental pathologies is rooted in past studies using epithelial Cre driver and knockout alleles. However, the available mouse models are limited, as most do not allow targeting different ameloblast sub-populations, and constitutive loss of EMPs often results in severe phenotype in the mineral, making it difficult to interpret defect mechanisms. Herein, we report on the design and verification of a toolkit of twelve mouse alleles that include ameloblast-stage specific Cre recombinases, fluorescent reporter alleles, and conditional flox alleles for the major EMPs. We show how these models may be used for applications such as sorting of live stage specific ameloblasts, whole mount imaging, and experiments with incisor explants. The full list of new alleles is available at https://dev.facebase.org/enamelatlas/mouse-models/.
]]></description>
<dc:creator>Wald, T.</dc:creator>
<dc:creator>Verma, A.</dc:creator>
<dc:creator>Cooley, V.</dc:creator>
<dc:creator>Marangoni, P.</dc:creator>
<dc:creator>Cazares, O.</dc:creator>
<dc:creator>Sharir, A.</dc:creator>
<dc:creator>Sandoval, E. J.</dc:creator>
<dc:creator>Sung, D.</dc:creator>
<dc:creator>Najibi, H.</dc:creator>
<dc:creator>Drennon, T. Y.</dc:creator>
<dc:creator>Bush, J. O.</dc:creator>
<dc:creator>Joester, D.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:date>2023-04-02</dc:date>
<dc:identifier>doi:10.1101/2023.03.30.534992</dc:identifier>
<dc:title><![CDATA[A Suite of Mouse Reagents for Studying Amelogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.03.534475v1?rss=1">
<title>
<![CDATA[
Paradoxical increases in anterior cingulate cortex activity during nitrous oxide-induced analgesia reveal a signature of pain affect 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.03.534475v1?rss=1</link>
<description><![CDATA[
The general consensus is that increases in neuronal activity in the anterior cingulate cortex (ACC) contribute to pains negative affect. Here, using in vivo imaging of neuronal calcium dynamics in mice, we report that nitrous oxide, a general anesthetic that reduces pain affect, paradoxically, increases ACC spontaneous activity. As expected, a noxious stimulus also increased ACC activity. However, as nitrous oxide increases baseline activity, the relative change in activity from pre-stimulus baseline was significantly less than the change in the absence of the general anesthetic. We suggest that this relative change in activity represents a neural signature of the affective pain experience. Furthermore, this signature of pain persists under general anesthesia induced by isoflurane, at concentrations in which the mouse is unresponsive. We suggest that this signature underlies the phenomenon of connected consciousness, in which use of the isolated forelimb technique revealed that pain percepts can persist in anesthetized patients.
]]></description>
<dc:creator>Weinrich, J. A.</dc:creator>
<dc:creator>Liu, C. D.</dc:creator>
<dc:creator>Jewell, M. E.</dc:creator>
<dc:creator>Andolina, C. R.</dc:creator>
<dc:creator>Bernstein, M. X.</dc:creator>
<dc:creator>Benitez, J.</dc:creator>
<dc:creator>Rodriguez-Rosado, S.</dc:creator>
<dc:creator>Braz, J. M.</dc:creator>
<dc:creator>Maze, M.</dc:creator>
<dc:creator>Nemenov, M. I.</dc:creator>
<dc:creator>Basbaum, A. I.</dc:creator>
<dc:date>2023-04-03</dc:date>
<dc:identifier>doi:10.1101/2023.04.03.534475</dc:identifier>
<dc:title><![CDATA[Paradoxical increases in anterior cingulate cortex activity during nitrous oxide-induced analgesia reveal a signature of pain affect]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.03.535288v1?rss=1">
<title>
<![CDATA[
High-throughput colocalization pipeline quantifies efficacy of mitochondrial targeting signals across different protein types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.03.535288v1?rss=1</link>
<description><![CDATA[
Efficient metabolic engineering and the development of mitochondrial therapeutics often rely upon the specific and strong import of foreign proteins into mitochondria. Fusing a protein to a mitochondria-bound signal peptide is a common method to localize proteins to mitochondria, but this strategy is not universally effective with particular proteins empirically failing to localize. To help overcome this barrier, this work develops a generalizable and open-source framework to design proteins for mitochondrial import and quantify their specific localization. By using a Python-based pipeline to quantitatively assess the colocalization of different proteins previously used for precise genome editing in a high-throughput manner, we reveal signal peptide-protein combinations that localize well in mitochondria and, more broadly, general trends about the overall reliability of commonly used mitochondrial targeting signals.
]]></description>
<dc:creator>Lear, S. K.</dc:creator>
<dc:creator>Nunez, J. A.</dc:creator>
<dc:creator>Shipman, S. L.</dc:creator>
<dc:date>2023-04-03</dc:date>
<dc:identifier>doi:10.1101/2023.04.03.535288</dc:identifier>
<dc:title><![CDATA[High-throughput colocalization pipeline quantifies efficacy of mitochondrial targeting signals across different protein types]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.30.534178v1?rss=1">
<title>
<![CDATA[
Computational drug repositioning for the identification of new agents to sensitize drug-resistant breast tumors across treatments and receptor subtypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.30.534178v1?rss=1</link>
<description><![CDATA[
Drug resistance is a major obstacle in cancer treatment and can involve a variety of different factors. Identifying effective therapies for drug resistant tumors is integral for improving patient outcomes. In this study, we applied a computational drug repositioning approach to identify potential agents to sensitize primary drug resistant breast cancers. We extracted drug resistance profiles from the I-SPY 2 TRIAL, a neoadjuvant trial for early stage breast cancer, by comparing gene expression profiles of responder and non-responder patients stratified into treatments within HR/HER2 receptor subtypes, yielding 17 treatment-subtype pairs. We then used a rank-based pattern-matching strategy to identify compounds in the Connectivity Map, a database of cell line derived drug perturbation profiles, that can reverse these signatures in a breast cancer cell line. We hypothesize that reversing these drug resistance signatures will sensitize tumors to treatment and prolong survival. We found that few individual genes are shared among the drug resistance profiles of different agents. At the pathway level, however, we found enrichment of immune pathways in the responders in 8 treatments within the HR+HER2+, HR+HER2-, and HR-HER2-receptor subtypes. We also found enrichment of estrogen response pathways in the non-responders in 10 treatments primarily within the hormone receptor positive subtypes. Although most of our drug predictions are unique to treatment arms and receptor subtypes, our drug repositioning pipeline identified the estrogen receptor antagonist fulvestrant as a compound that can potentially reverse resistance across 13/17 of the treatments and receptor subtypes including HR+ and triple negative. While fulvestrant showed limited efficacy when tested in a panel of 5 paclitaxel-resistant breast cancer cell lines, it did increase drug response in combination with paclitaxel in HCC-1937, a triple negative breast cancer cell line.
]]></description>
<dc:creator>Yu, K.</dc:creator>
<dc:creator>Basu, A.</dc:creator>
<dc:creator>Yau, C.</dc:creator>
<dc:creator>Wolf, D. M.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Bandyopadhyay, S.</dc:creator>
<dc:creator>Korkola, J. E.</dc:creator>
<dc:creator>Hirst, G. L.</dc:creator>
<dc:creator>Asare, S.</dc:creator>
<dc:creator>I-SPY 2 TRIAL,</dc:creator>
<dc:creator>Demichele, A.</dc:creator>
<dc:creator>Hylton, N.</dc:creator>
<dc:creator>Yee, D.</dc:creator>
<dc:creator>Esserman, L.</dc:creator>
<dc:creator>van 't Veer, L.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2023-04-04</dc:date>
<dc:identifier>doi:10.1101/2023.03.30.534178</dc:identifier>
<dc:title><![CDATA[Computational drug repositioning for the identification of new agents to sensitize drug-resistant breast tumors across treatments and receptor subtypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.03.535302v1?rss=1">
<title>
<![CDATA[
Piezo inhibition prevents and rescues scarring by targeting the adipocyte to fibroblast transition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.03.535302v1?rss=1</link>
<description><![CDATA[
While past studies have suggested that plasticity exists between dermal fibroblasts and adipocytes, it remains unknown whether fat actively contributes to fibrosis in scarring. We show that adipocytes convert to scar-forming fibroblasts in response to Piezo-mediated mechanosensing to drive wound fibrosis. We establish that mechanics alone are sufficient to drive adipocyte-to- fibroblast conversion. By leveraging clonal-lineage-tracing in combination with scRNA-seq, Visium, and CODEX, we define a "mechanically naive" fibroblast-subpopulation that represents a transcriptionally intermediate state between adipocytes and scar-fibroblasts. Finally, we show that Piezo1 or Piezo2-inhibition yields regenerative healing by preventing adipocytes activation to fibroblasts, in both mouse-wounds and a novel human-xenograft-wound model. Importantly, Piezo1-inhibition induced wound regeneration even in pre-existing established scars, a finding that suggests a role for adipocyte-to-fibroblast transition in wound remodeling, the least-understood phase of wound healing. Adipocyte-to-fibroblast transition may thus represent a therapeutic target for minimizing fibrosis via Piezo-inhibition in organs where fat contributes to fibrosis.
]]></description>
<dc:creator>Griffin, M.</dc:creator>
<dc:creator>Talbott, H.</dc:creator>
<dc:creator>Guardino, N.</dc:creator>
<dc:creator>Guo, J.</dc:creator>
<dc:creator>Spielman, A.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Mascharak, S.</dc:creator>
<dc:creator>Parker, J.</dc:creator>
<dc:creator>Henn, D.</dc:creator>
<dc:creator>Liang, N.</dc:creator>
<dc:creator>King, M.</dc:creator>
<dc:creator>Cotterell, A.</dc:creator>
<dc:creator>Bauer-Rowe, K.</dc:creator>
<dc:creator>Abbas, D.</dc:creator>
<dc:creator>Deleon, N. M. D.</dc:creator>
<dc:creator>Fahy, E.</dc:creator>
<dc:creator>Sivaraj, D.</dc:creator>
<dc:creator>Downer, M.</dc:creator>
<dc:creator>Akras, D.</dc:creator>
<dc:creator>Berry, C.</dc:creator>
<dc:creator>Cook, J.</dc:creator>
<dc:creator>Quarto, N.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Lorenz, P.</dc:creator>
<dc:creator>Gurtner, G.</dc:creator>
<dc:creator>Januszyk, M.</dc:creator>
<dc:creator>Wan, D. C.</dc:creator>
<dc:creator>Longaker, M. T.</dc:creator>
<dc:date>2023-04-04</dc:date>
<dc:identifier>doi:10.1101/2023.04.03.535302</dc:identifier>
<dc:title><![CDATA[Piezo inhibition prevents and rescues scarring by targeting the adipocyte to fibroblast transition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.04.535480v1?rss=1">
<title>
<![CDATA[
Comparing chromatin contact maps at scale: methods and insights 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.04.535480v1?rss=1</link>
<description><![CDATA[
Comparing chromatin contact maps is an essential step in quantifying how three-dimensional (3D) genome organization shapes development, evolution, and disease. However, no gold standard exists for comparing contact maps, and even simple methods often disagree. In this study, we propose novel comparison methods and evaluate them alongside existing approaches using genome-wide Hi-C data and 22,500 in silico predicted contact maps. We also quantify the robustness of methods to common sources of biological and technical variation, such as boundary size and noise. We find that simple difference-based methods such as mean squared error are suitable for initial screening, but biologically informed methods are necessary to identify why maps diverge and propose specific functional hypotheses. We provide a reference guide, codebase, and benchmark for rapidly comparing chromatin contact maps at scale to enable biological insights into the 3D organization of the genome.
]]></description>
<dc:creator>Gunsalus, L. M.</dc:creator>
<dc:creator>McArthur, E.</dc:creator>
<dc:creator>Gjoni, K.</dc:creator>
<dc:creator>Kuang, S.</dc:creator>
<dc:creator>Pittman, M.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2023-04-04</dc:date>
<dc:identifier>doi:10.1101/2023.04.04.535480</dc:identifier>
<dc:title><![CDATA[Comparing chromatin contact maps at scale: methods and insights]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.04.535379v1?rss=1">
<title>
<![CDATA[
Spatial analysis of human lung cancer reveals organized immune hubs enriched for stem-like CD8 T cells and associated with immunotherapy response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.04.535379v1?rss=1</link>
<description><![CDATA[
The organization of immune cells in human tumors is not well understood. Immunogenic tumors harbor spatially-localized multicellular  immunity hubs defined by expression of the T cell-attracting chemokines CXCL10/CXCL11 and abundant T cells. Here, we examined immunity hubs in human pre-immunotherapy lung cancer specimens, and found that they were associated with beneficial responses to PD-1-blockade. Immunity hubs were enriched for many interferon-stimulated genes, T cells in multiple differentiation states, and CXCL9/10/11+ macrophages that preferentially interact with CD8 T cells. Critically, we discovered the stem-immunity hub, a subtype of immunity hub strongly associated with favorable PD-1-blockade outcomes, distinct from mature tertiary lymphoid structures, and enriched for stem-like TCF7+PD-1+ CD8 T cells and activated CCR7+LAMP3+ dendritic cells, as well as chemokines that organize these cells. These results elucidate the spatial organization of the human intratumoral immune response and its relevance to patient immunotherapy outcomes.
]]></description>
<dc:creator>Chen, J. H.</dc:creator>
<dc:creator>Nieman, L. T.</dc:creator>
<dc:creator>Spurrell, M.</dc:creator>
<dc:creator>Jorgji, V.</dc:creator>
<dc:creator>Richieri, P. M.</dc:creator>
<dc:creator>Xu, K. H.</dc:creator>
<dc:creator>Madhu, R.</dc:creator>
<dc:creator>Parikh, M.</dc:creator>
<dc:creator>Zamora, I.</dc:creator>
<dc:creator>Mehta, A.</dc:creator>
<dc:creator>Nabel, C. S.</dc:creator>
<dc:creator>Freeman, S. S.</dc:creator>
<dc:creator>Pirl, J. D.</dc:creator>
<dc:creator>Lu, C.</dc:creator>
<dc:creator>Meador, C. B.</dc:creator>
<dc:creator>Barth, J.</dc:creator>
<dc:creator>Sakhi, M.</dc:creator>
<dc:creator>Tang, A. L.</dc:creator>
<dc:creator>Sarkizova, S.</dc:creator>
<dc:creator>Price, C.</dc:creator>
<dc:creator>Yizhak, K.</dc:creator>
<dc:creator>Hofree, M.</dc:creator>
<dc:creator>Shih, A.</dc:creator>
<dc:creator>Sade-Feldman, M.</dc:creator>
<dc:creator>Boland, G. M.</dc:creator>
<dc:creator>Pelka, K.</dc:creator>
<dc:creator>Aryee, M. J.</dc:creator>
<dc:creator>Korsunsky, I.</dc:creator>
<dc:creator>Mino-Kenudson, M.</dc:creator>
<dc:creator>Gainor, J.</dc:creator>
<dc:creator>Hacohen, N.</dc:creator>
<dc:date>2023-04-06</dc:date>
<dc:identifier>doi:10.1101/2023.04.04.535379</dc:identifier>
<dc:title><![CDATA[Spatial analysis of human lung cancer reveals organized immune hubs enriched for stem-like CD8 T cells and associated with immunotherapy response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.04.535452v1?rss=1">
<title>
<![CDATA[
Revealing the Grammar of Small RNA Secretion Using Interpretable Machine Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.04.535452v1?rss=1</link>
<description><![CDATA[
Small non-coding RNAs can be secreted through a variety of mechanisms, including exosomal sorting, in small extracellular vesicles, and within lipoprotein complexes 1,2. However, the mechanisms that govern their sorting and secretion are still not well understood. In this study, we present ExoGRU, a machine learning model that predicts small RNA secretion probabilities from primary RNA sequence. We experimentally validated the performance of this model through ExoGRU-guided mutagenesis and synthetic RNA sequence analysis, and confirmed that primary RNA sequence is a major determinant in small RNA secretion. Additionally, we used ExoGRU to reveal cis and trans factors that underlie small RNA secretion, including known and novel RNA-binding proteins, e.g., YBX1, HNRNPA2B1, and RBM24. We also developed a novel technique called exoCLIP, which reveals the RNA interactome of RBPs within the cell-free space. We used exoCLIP to reveal the RNA interactome of HNRNPA2B1 and RBM24 in extracellular vesicles. Together, our results demonstrate the power of machine learning in revealing novel biological mechanisms. In addition to providing deeper insight into complex processes such as small RNA secretion, this knowledge can be leveraged in therapeutic and synthetic biology applications.
]]></description>
<dc:creator>Zirak, B.</dc:creator>
<dc:creator>Naghipourfar, M.</dc:creator>
<dc:creator>Saberi, A.</dc:creator>
<dc:creator>Pouyabahar, D.</dc:creator>
<dc:creator>Zarezadeh, A.</dc:creator>
<dc:creator>Fish, L.</dc:creator>
<dc:creator>Luo, L.</dc:creator>
<dc:creator>Huh, D.</dc:creator>
<dc:creator>Sharifi-Zarchi, A.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2023-04-06</dc:date>
<dc:identifier>doi:10.1101/2023.04.04.535452</dc:identifier>
<dc:title><![CDATA[Revealing the Grammar of Small RNA Secretion Using Interpretable Machine Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.06.535777v1?rss=1">
<title>
<![CDATA[
Parallelized screening of virus accessory genes reveals diverse defense and counter-defense mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.06.535777v1?rss=1</link>
<description><![CDATA[
Viruses have been evolving host-modifying factors for billions of years. Genomes of bacterial and archaeal viruses are replete with fast-evolving, uncharacterized accessory genes (AGs), most of which likely antagonize host defenses or other viruses1, 2. Systematic investigation of AGs could uncover a multitude of biological mechanisms involved in virus-host competition, but AG identification in genomic databases remains a challenge. We developed an integrated computational and high-throughput discovery platform to identify AGs in virus genomes and assay their functions in complementary phage infection-dependent and -independent contexts. Our approach showcases how phages interact with the principal layers of antiviral immunity, including cell surface modifications, restriction systems, and abortive infection (Abi) mechanisms, which operate simultaneously in the same host. We discovered multiple Enterobacteriophage AGs associated with counter-defense functions that activate rather than inhibit antiviral immunity in cells, including the surprising finding that anti-restriction AGs elicit programmed cell death (PCD) activity of some restriction-modification (R-M) systems. We propose that counter-defense AGs that trigger PCD create a conundrum for phages whereby keeping the AGs causes PCD but losing them exposes the phage to restriction by bacteria. Strategies employed by viruses to avoid this double jeopardy could be an important factor in virus evolution that remains to be explored.
]]></description>
<dc:creator>Silas, S.</dc:creator>
<dc:creator>Carion, H.</dc:creator>
<dc:creator>Makarova, K. S.</dc:creator>
<dc:creator>Laderman, E.</dc:creator>
<dc:creator>Godinez, D. S.</dc:creator>
<dc:creator>Johnson, M.</dc:creator>
<dc:creator>Fossati, A.</dc:creator>
<dc:creator>Swaney, D.</dc:creator>
<dc:creator>Bocek, M.</dc:creator>
<dc:creator>Koonin, E. V.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2023-04-06</dc:date>
<dc:identifier>doi:10.1101/2023.04.06.535777</dc:identifier>
<dc:title><![CDATA[Parallelized screening of virus accessory genes reveals diverse defense and counter-defense mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.06.535844v1?rss=1">
<title>
<![CDATA[
Progranulin inhibits phospholipase sPLA2-IIA to control neuroinflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.06.535844v1?rss=1</link>
<description><![CDATA[
Withdrawal StatementThe authors have withdrawn this manuscript because more work is needed to fully define the role of sPLA2-IIA. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.
]]></description>
<dc:creator>Du, H.</dc:creator>
<dc:creator>Yang, C.</dc:creator>
<dc:creator>Nana, A. L.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Smolka, M. B.</dc:creator>
<dc:creator>Hu, F.</dc:creator>
<dc:date>2023-04-06</dc:date>
<dc:identifier>doi:10.1101/2023.04.06.535844</dc:identifier>
<dc:title><![CDATA[Progranulin inhibits phospholipase sPLA2-IIA to control neuroinflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.07.536070v1?rss=1">
<title>
<![CDATA[
A critical role for PLCG1 in RAS activation by BCR-ABL1 and FLT3-ITD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.07.536070v1?rss=1</link>
<description><![CDATA[
Myeloid leukemias are frequently associated with pathologically activating mutations in tyrosine kinases [BCR-ABL1 in chronic myeloid leukemia (CML); FLT3 juxtamembrane internal tandem duplication (ITD) mutations, FLT3 and KIT activation loop mutations in acute myeloid leukemia (AML)]. Mutations in these kinases activate RAS, which initiates multiple downstream signaling pathways that regulate cell proliferation, differentiation, and apoptosis. The mechanisms whereby RAS is activated by these kinases is incompletely understood, and a better understanding of the molecular mediators involved in RAS activation may uncover new therapeutic strategies. Here we identify a biologically and therapeutically important novel mechanism whereby BCR-ABL1 and FLT3-ITD activate the critical downstream effector RAS in part through phospholipase C gamma-1 (PLCG1). PLCG1 knockout decreases proliferation of CML and FLT3-ITD-expressing AML cells, reduces RAS nucleotide exchange factor activity, and increases sensitivity of CML cells to BCR-ABL1 tyrosine kinase inhibitors (TKIs). Collectively, these studies suggest that PLCG1 inhibition may augment clinical responses to BCR-ABL1 and FLT3 TKIs in CML and AML.
]]></description>
<dc:creator>Szeto, M. Y.</dc:creator>
<dc:creator>Mase, A.</dc:creator>
<dc:creator>Kulhanek, K.</dc:creator>
<dc:creator>Banerjee, S.</dc:creator>
<dc:creator>Rubio, I.</dc:creator>
<dc:creator>Roose, J.</dc:creator>
<dc:creator>Shah, N. P.</dc:creator>
<dc:date>2023-04-10</dc:date>
<dc:identifier>doi:10.1101/2023.04.07.536070</dc:identifier>
<dc:title><![CDATA[A critical role for PLCG1 in RAS activation by BCR-ABL1 and FLT3-ITD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.10.536290v1?rss=1">
<title>
<![CDATA[
Secreted HLA Fc-Fusion Profiles Immunopeptidome in Hypoxic PDAC and Cellular Senescence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.10.536290v1?rss=1</link>
<description><![CDATA[
Human leukocyte antigens (HLA) display peptides largely from intracellular proteins on the surface of cells in major histocompatibility complex (MHC)-peptide complexes. These complexes provide a biological window into the cell, and peptides derived from disease-associated antigens can serve as biomarkers and therapeutic targets. Thus, proper identification of peptides and the corresponding presenting HLA allele in disease phenotypes is important for the design and execution of therapeutic strategies using engineered T-cell receptors or antibodies. Yet, current mass spectrometry methods for profiling the immunopeptidome typically require large and complex sample inputs, complicating the study of several disease phenotypes and lowering the confidence of both peptide and allele identification. Here, we describe a novel secreted HLA (sHLA) Fc-fusion construct that allows for simple peptide identification from single HLA alleles in two important disease models: hypoxic pancreatic ductal adenocarcinoma (PDAC) and cellular senescence. We identify hypoxia and senescence-associated peptides that could act as future targets for immunotherapy. More generally, the method streamlines the time between sample preparation and injection from days to hours, yielding allele-restricted target identification in a temporally controlled manner. Overall, this method identified >30,000 unique HLA-associated peptides across two different HLA alleles and seven cell lines. Notably, [~]9,300 of these unique HLA-associated peptides had previously not been identified in the Immune Epitope Database. We believe the sHLA Fc-fusion capture technology will accelerate the study of the immunopeptidome as therapeutic interest in HLA-peptide complexes increases in cancer and beyond.
]]></description>
<dc:creator>Rettko, N. J.</dc:creator>
<dc:creator>Kirkemo, L. L.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2023-04-12</dc:date>
<dc:identifier>doi:10.1101/2023.04.10.536290</dc:identifier>
<dc:title><![CDATA[Secreted HLA Fc-Fusion Profiles Immunopeptidome in Hypoxic PDAC and Cellular Senescence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.11.536275v1?rss=1">
<title>
<![CDATA[
Demultiplex2: robust sample demultiplexing for scRNA-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.11.536275v1?rss=1</link>
<description><![CDATA[
Single-cell sample multiplexing technologies function by associating sample-specific barcode tags with cell-specific barcode tags, thereby increasing sample throughput, reducing batch effects, and decreasing reagent costs. Computational methods must then correctly associate cell-tags with sample-tags, but their performance deteriorates rapidly when working with datasets that are large, have imbalanced cell numbers across samples, or are noisy due to cross-contamination among sample tags - unavoidable features of many real-world experiments. Here we introduce deMULTIplex2, a mechanism-guided classification algorithm for multiplexed scRNA-seq data that successfully recovers many more cells across a spectrum of challenging datasets compared to existing methods. deMULTIplex2 is built on a statistical model of tag read counts derived from the physical mechanism of tag cross-contamination. Using generalized linear models and expectation-maximization, deMULTIplex2 probabilistically infers the sample identity of each cell and classifies singlets with high accuracy. Using Randomized Quantile Residuals, we show the model fits both simulated and real datasets. Benchmarking analysis suggests that deMULTIplex2 outperforms existing algorithms, especially when handling large and noisy single-cell datasets or those with unbalanced sample compositions.
]]></description>
<dc:creator>Zhu, Q.</dc:creator>
<dc:creator>Conrad, D. N.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:date>2023-04-12</dc:date>
<dc:identifier>doi:10.1101/2023.04.11.536275</dc:identifier>
<dc:title><![CDATA[Demultiplex2: robust sample demultiplexing for scRNA-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.11.536358v1?rss=1">
<title>
<![CDATA[
An automated proximity proteomics pipeline for subcellular proteome and protein interaction mapping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.11.536358v1?rss=1</link>
<description><![CDATA[
Proximity labeling (PL) through biotinylation coupled with mass spectrometry (MS) has emerged as a powerful technique for capturing spatial proteomes within living cells. Large-scale sample processing for proximity proteomics requires a workflow that minimizes hands-on time while enhancing quantitative reproducibility. Here, we present a scalable PL pipeline integrating automated enrichment of biotinylated proteins in a 96-well plate format. By combining this pipeline with an optimized quantitative MS acquisition method based on data-independent acquisition (DIA), we not only significantly increased sample throughput but also improved the reproducibility of protein identification and quantification. We applied this pipeline to delineate subcellular proteomes across various cellular compartments, including endosomes, late endosomes/lysosomes, the Golgi apparatus, and the plasma membrane. Moreover, employing 5HT2A serotonin receptor as a model, we investigated temporal changes of proximal interaction networks induced by the receptors activation with serotonin. Finally, to demonstrate the applicability of our PL pipeline across multiple experimental conditions, we further modified the PL pipeline for reduced sample input amounts to accommodate CRISPR-based gene knockout, and assessed the dynamics of the 5HT2A network in response to the perturbation of selected proximal interactors. Importantly, the presented PL approach is universally applicable to PL proteomics using biotinylation-based PL enzymes, increasing both throughput and reproducibility of standard protocols.
]]></description>
<dc:creator>Huttenhain, R.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Eckhardt, M.</dc:creator>
<dc:creator>von Zastrow, M.</dc:creator>
<dc:creator>Roth, B.</dc:creator>
<dc:creator>Foussard, H.</dc:creator>
<dc:creator>Marley, A.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Vartak, R.</dc:creator>
<dc:creator>DiBerto, J. F.</dc:creator>
<dc:creator>Patil, T.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Zhong, X.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:date>2023-04-12</dc:date>
<dc:identifier>doi:10.1101/2023.04.11.536358</dc:identifier>
<dc:title><![CDATA[An automated proximity proteomics pipeline for subcellular proteome and protein interaction mapping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.12.536497v1?rss=1">
<title>
<![CDATA[
Deletion of Pax1 scoliosis-associated regulatory elements leads to a female-biased tail abnormality 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.12.536497v1?rss=1</link>
<description><![CDATA[
Adolescent idiopathic scoliosis (AIS), a sideways curvature of the spine, is sexually dimorphic, with increased incidence in females. A GWAS identified a female-specific AIS susceptibility locus near the PAX1 gene. Here, we used mouse enhancer assays, three mouse enhancer knockouts and subsequent phenotypic analyses to characterize this region. Using mouse enhancer assays, we characterized a sequence, PEC7, that overlaps the AIS-associated variant, and found it to be active in the tail tip and intervertebral disc. Removal of PEC7 or Xe1, a known sclerotome enhancer nearby, and deletion of both sequences led to a kinky phenotype only in the Xe1 and combined (Xe1+PEC7) knockouts, with only the latter showing a female sex dimorphic phenotype. Extensive phenotypic characterization of these mouse lines implicated several differentially expressed genes and estrogen signaling in the sex dimorphic bias. In summary, our work functionally characterizes an AIS-associated locus and dissects the mechanism for its sexual dimorphism.
]]></description>
<dc:creator>Ushiki, A.</dc:creator>
<dc:creator>Sheng, R. R.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Nobuhara, M.</dc:creator>
<dc:creator>Murray, E.</dc:creator>
<dc:creator>Ruan, X.</dc:creator>
<dc:creator>Rios, J. J.</dc:creator>
<dc:creator>Wise, C. A.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2023-04-13</dc:date>
<dc:identifier>doi:10.1101/2023.04.12.536497</dc:identifier>
<dc:title><![CDATA[Deletion of Pax1 scoliosis-associated regulatory elements leads to a female-biased tail abnormality]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.12.536655v1?rss=1">
<title>
<![CDATA[
Non-specific recognition of histone modifications by H3K9bhb antibody 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.12.536655v1?rss=1</link>
<description><![CDATA[
Ketone bodies are short chain fatty acids produced in the liver during periods of limited glucose availability that provide an alternative source of energy for the brain, heart, and skeletal muscle. Beyond this classical metabolic role, {beta}-hydroxybutyrate (BHB), is gaining recognition as a pleiotropic signaling molecule. Lysine {beta}-hydroxybutyrylation (Kbhb) is a newly discovered post-translational modification in which BHB is covalently attached to lysine {varepsilon}-amino groups. This novel protein adduct is metabolically sensitive, dependent on BHB concentration, and found on proteins in multiple intracellular compartments, including the mitochondria and nucleus. Therefore, Kbhb is hypothesized to be an important component of ketone body-regulated physiology. Kbhb on histones is proposed to be an epigenetic regulator, which links metabolic alterations to gene expression. However, we found that the widely used antibody against the {beta}-hydroxybutyrylated lysine 9 on histone H3 (H3K9bhb) also recognizes other modification(s), which are increased by deacetylation inhibition and include likely acetylations. Therefore, caution must be used when interpreting gene regulation data acquired with the H3K9bhb antibody.
]]></description>
<dc:creator>Tsusaka, T.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>DeFelice, B.</dc:creator>
<dc:creator>Burlingame, A.</dc:creator>
<dc:creator>Goldberg, E. L.</dc:creator>
<dc:date>2023-04-13</dc:date>
<dc:identifier>doi:10.1101/2023.04.12.536655</dc:identifier>
<dc:title><![CDATA[Non-specific recognition of histone modifications by H3K9bhb antibody]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.13.536796v1?rss=1">
<title>
<![CDATA[
SEL1L-HRD1 ER-associated degradation is a new ataxia gene 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.13.536796v1?rss=1</link>
<description><![CDATA[
The SEL1L-HRD1 protein complex represents the most conserved branch of endoplasmic reticulum (ER)-associated degradation (ERAD); however, definitive evidence for the importance of SEL1L in HRD1 ERAD is lacking. Here we report that attenuation of the interaction between SEL1L and HRD1 impairs HRD1 ERAD function and has pathological consequences in mice. Our data show that SEL1L variant p.Ser658Pro (SEL1LS658P) previously identified in Finnish Hound suffering cerebellar ataxia is a recessive hypomorphic mutation, causing partial embryonic lethality, developmental delay, and early-onset cerebellar ataxia in homozygous mice carrying the bi-allelic variant. Mechanistically, SEL1LS658P variant attenuates the SEL1L-HRD1 interaction and causes HRD1 dysfunction by generating electrostatic repulsion between SEL1L F668 and HRD1 Y30 residues. Proteomic screens of SEL1L and HRD1 interactomes revealed that the SEL1L-HRD1 interaction is prerequisite for the formation of a functional HRD1 ERAD complex, as SEL1L recruits not only the lectins OS9 and ERLEC1, but the E2 UBE2J1 and retrotranslocon DERLIN, to HRD1. These data underscore the pathophysiological importance and disease relevance of the SEL1L-HRD1 complex, and identify a key step in organizing the HRD1 ERAD complex.
]]></description>
<dc:creator>Lin, L. L.</dc:creator>
<dc:creator>Torres, M.</dc:creator>
<dc:creator>Pederson, B.</dc:creator>
<dc:creator>Wang, H. H.</dc:creator>
<dc:creator>Wei, X.</dc:creator>
<dc:creator>Li, Z. J.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Mao, H.</dc:creator>
<dc:creator>Hanzel, M.</dc:creator>
<dc:creator>Govek, E. E.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Hatten, M. E.</dc:creator>
<dc:creator>Sun, S. E.</dc:creator>
<dc:creator>Qi, L.</dc:creator>
<dc:date>2023-04-14</dc:date>
<dc:identifier>doi:10.1101/2023.04.13.536796</dc:identifier>
<dc:title><![CDATA[SEL1L-HRD1 ER-associated degradation is a new ataxia gene]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.14.536909v1?rss=1">
<title>
<![CDATA[
Total Chemical Synthesis of Glycosylated TREM2 Ectodomain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.14.536909v1?rss=1</link>
<description><![CDATA[
Mutations in a microglia-associated gene TREM2 increase the risk of Alzheimers disease. Currently, structural and functional studies of TREM2 mainly rely on recombinant TREM2 proteins expressed from mammalian cells. However, using this method, it is difficult to achieve site-specific labeling. Here we present the total chemical synthesis of the 116 amino-acid TREM2 ectodomain. Rigorous structural analysis ensured correct structural fold after refolding. Treating microglial cells with refolded synthetic TREM2 enhanced microglial phagocytosis, proliferation, and survival. We also prepared TREM2 constructs with defined glycosylation patterns and found that glycosylation at N79 is critical to the thermal stability of TREM2. This method will provide access to TREM2 constructs with site-specific labeling, such as fluorescent labeling, reactive chemical handles, and enrichment handles, to further advance our understanding of TREM2 in Alzheimers disease.
]]></description>
<dc:creator>Wijegunawardena, G.</dc:creator>
<dc:creator>Castillo, E.</dc:creator>
<dc:creator>Henrickson, B.</dc:creator>
<dc:creator>Davis, R.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:date>2023-04-14</dc:date>
<dc:identifier>doi:10.1101/2023.04.14.536909</dc:identifier>
<dc:title><![CDATA[Total Chemical Synthesis of Glycosylated TREM2 Ectodomain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.14.535921v1?rss=1">
<title>
<![CDATA[
Integrative identification of non-coding regulatory regions driving metastatic prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.14.535921v1?rss=1</link>
<description><![CDATA[
Large-scale sequencing efforts of thousands of tumor samples have been undertaken to understand the mutational landscape of the coding genome. However, the vast majority of germline and somatic variants occur within non-coding portions of the genome. These genomic regions do not directly encode for specific proteins, but can play key roles in cancer progression, for example by driving aberrant gene expression control. Here, we designed an integrative computational and experimental framework to identify recurrently mutated non-coding regulatory regions that drive tumor progression. Application of this approach to whole-genome sequencing (WGS) data from a large cohort of metastatic castration-resistant prostate cancer (mCRPC) revealed a large set of recurrently mutated regions. We used (i) in silico prioritization of functional non-coding mutations, (ii) massively parallel reporter assays, and (iii) in vivo CRISPR-interference (CRISPRi) screens in xenografted mice to systematically identify and validate driver regulatory regions that drive mCRPC. We discovered that one of these enhancer regions, GH22I030351, acts on a bidirectional promoter to simultaneously modulate expression of U2-associated splicing factor SF3A1 and chromosomal protein CCDC157. We found that both SF3A1 and CCDC157 are promoters of tumor growth in xenograft models of prostate cancer. We nominated a number of transcription factors, including SOX6, to be responsible for higher expression of SF3A1 and CCDC157. Collectively, we have established and confirmed an integrative computational and experimental approach that enables the systematic detection of non-coding regulatory regions that drive the progression of human cancers.
]]></description>
<dc:creator>Woo, B. J.</dc:creator>
<dc:creator>Moussavi-Baygi, R.</dc:creator>
<dc:creator>Karner, H.</dc:creator>
<dc:creator>Karimzadeh, M.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Joshi, T.</dc:creator>
<dc:creator>Yin, K.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Asgharian, H.</dc:creator>
<dc:creator>Feng, F. Y.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2023-04-16</dc:date>
<dc:identifier>doi:10.1101/2023.04.14.535921</dc:identifier>
<dc:title><![CDATA[Integrative identification of non-coding regulatory regions driving metastatic prostate cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.16.537084v1?rss=1">
<title>
<![CDATA[
Axon initial segment GABA inhibits action potential generation throughout periadolescent development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.16.537084v1?rss=1</link>
<description><![CDATA[
Neurons are remarkably polarized structures: dendrites spread and branch to receive synaptic inputs while a single axon extends and transmits action potentials to downstream targets. Neuronal polarity is maintained by the axon initial segment (AIS), a region between the soma and axon proper that is also the site of action potential (AP) generation. This polarization between dendrites and axons extends to inhibitory neurotransmission. In adulthood, the neurotransmitter GABA hyperpolarizes dendrites but instead depolarizes axons. These differences in function collide at the AIS. Multiple studies have shown that GABAergic signaling in this region can share properties of either the mature axon or mature dendrite, and that these properties evolve over a protracted period encompassing periadolescent development. Here, we explored how developmental changes in GABAergic signaling affect AP initiation. We show that GABA at the axon initial segment inhibits action potential initiation in Layer 2/3 pyramidal neurons in prefrontal cortex from mice of either sex across GABA reversal potentials observed in periadolescence. These actions occur largely through current shunts generated by GABAA receptors and changes in voltage-gated channel properties that affected the number of channels that could be recruited for AP electrogenesis. These results suggest that GABAergic neurons targeting the axon initial segment provide an inhibitory "veto" across the range of GABA polarity observed in normal adolescent development, regardless of GABAergic synapse reversal potential.
]]></description>
<dc:creator>Lipkin, A. M.</dc:creator>
<dc:creator>Bender, K. J.</dc:creator>
<dc:date>2023-04-16</dc:date>
<dc:identifier>doi:10.1101/2023.04.16.537084</dc:identifier>
<dc:title><![CDATA[Axon initial segment GABA inhibits action potential generation throughout periadolescent development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.14.536843v1?rss=1">
<title>
<![CDATA[
The lncRNA Malat1 Inhibits miR-15/16 to Enhance Cytotoxic T Cell Activation and Memory Cell Formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.14.536843v1?rss=1</link>
<description><![CDATA[
Proper activation of cytotoxic T cells via the T cell receptor and the costimulatory receptor CD28 is essential for adaptive immunity against viruses, many intracellular bacteria and cancers. Through biochemical analysis of RNA:protein interactions, we uncovered a non-coding RNA circuit regulating activation and differentiation of cytotoxic T cells composed of the long non-coding RNA Malat1 (Metastasis Associated Lung Adenocarcinoma Transcript 1) and the microRNA family miR-15/16. miR-15/16 is a widely and highly expressed tumor suppressor miRNA family important for cell proliferation and survival. miR-15/16 also play important roles in T cell responses to viral infection, including the regulation of antigen-specific T cell expansion and T cell memory. Comparative Argonaute-2 high throughput sequencing of crosslinking immunoprecipitation (Ago2 HITS-CLIP, or AHC) combined with gene expression profiling in normal and miR-15/16-deficient T cells revealed a large network of several hundred direct miR-15/16 target mRNAs, many with functional relevance for T cell activation, survival and memory formation. Among these targets, the long non-coding RNA Malat1 contained the largest absolute magnitude miR-15/16-dependent AHC peak in T cells. This binding site was also among the strongest lncRNA:miRNA interactions detected in the T cell transcriptome. We used CRISPR targeting with homology directed repair to generate mice with a 5-nucleotide mutation in the miR-15/16 binding site in Malat1. This mutation interrupted Malat1:miR-15/16 interaction, and enhanced the repression of other miR-15/16 target genes, including CD28. Interrupting Malat1 interaction with miR-15/16 decreased cytotoxic T cell activation, including the expression of IL-2 and a broader CD28-responsive gene program. Accordingly, Malat1 mutation diminished memory cell persistence following LCMV Armstrong and Listeria monocytogenes infection. This study marks a significant advance in the study of long noncoding RNAs in the immune system by ascribing cell-intrinsic, sequence-specific in vivo function to Malat1. These findings have implications for T cell-mediated autoimmune diseases, antiviral and anti-tumor immunity, as well as lung adenocarcinoma and other malignancies where Malat1 is overexpressed.



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]]></description>
<dc:creator>Wheeler, B. D.</dc:creator>
<dc:creator>Gagnon, J. D.</dc:creator>
<dc:creator>Zhu, W. S.</dc:creator>
<dc:creator>Munoz-Sandoval, P.</dc:creator>
<dc:creator>Wong, S. K.</dc:creator>
<dc:creator>Simeonov, D. S.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Debarge, R.</dc:creator>
<dc:creator>Spitzer, M.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Ansel, K. M.</dc:creator>
<dc:date>2023-04-17</dc:date>
<dc:identifier>doi:10.1101/2023.04.14.536843</dc:identifier>
<dc:title><![CDATA[The lncRNA Malat1 Inhibits miR-15/16 to Enhance Cytotoxic T Cell Activation and Memory Cell Formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.17.536878v1?rss=1">
<title>
<![CDATA[
Finding the right tool: a comprehensive evaluation of microglial inducible cre mouse models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.17.536878v1?rss=1</link>
<description><![CDATA[
The recent proliferation of new Cre and CreER recombinase lines provides researchers with a diverse toolkit to study microglial gene function. To determine how best to apply these lines in studies of microglial gene function, a thorough and detailed comparison of their properties is needed. Here, we examined four different microglial CreER lines (Cx3cr1CreER(Litt), Cx3cr1CreER(Jung), P2ry12CreER, Tmem119CreER), focusing on (1) recombination specificity; (2) leakiness - degree of non-tamoxifen recombination in microglia and other cells; (3) efficiency of tamoxifen-induced recombination; (4) extra-neural recombination -the degree of recombination in cells outside the CNS, particularly myelo/monocyte lineages (5) off-target effects in the context of neonatal brain development. We identify important caveats and strengths for these lines which will provide broad significance for researchers interested in performing conditional gene deletion in microglia. We also provide data emphasizing the potential of these lines for injury models that result in the recruitment of splenic immune cells.
]]></description>
<dc:creator>Bedolla, A. M.</dc:creator>
<dc:creator>McKinsey, G.</dc:creator>
<dc:creator>Ware, K.</dc:creator>
<dc:creator>Santander, N.</dc:creator>
<dc:creator>Arnold, T.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:date>2023-04-17</dc:date>
<dc:identifier>doi:10.1101/2023.04.17.536878</dc:identifier>
<dc:title><![CDATA[Finding the right tool: a comprehensive evaluation of microglial inducible cre mouse models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.18.537104v1?rss=1">
<title>
<![CDATA[
A single inactivating amino acid change in the SARS-CoV-2 NSP3 Mac1 domain attenuates viral replication and pathogenesis in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.18.537104v1?rss=1</link>
<description><![CDATA[
Despite unprecedented efforts, our therapeutic arsenal against SARS-CoV-2 remains limited. The conserved macrodomain 1 (Mac1) in NSP3 is an enzyme exhibiting ADP-ribosylhydrolase activity and a possible drug target. To determine the therapeutic potential of Mac1 inhibition, we generated recombinant viruses and replicons encoding a catalytically inactive NSP3 Mac1 domain by mutating a critical asparagine in the active site. While substitution to alanine (N40A) reduced catalytic activity by [~]10-fold, mutations to aspartic acid (N40D) reduced activity by [~]100-fold relative to wildtype. Importantly, the N40A mutation rendered Mac1 unstable in vitro and lowered expression levels in bacterial and mammalian cells. When incorporated into SARS-CoV-2 molecular clones, the N40D mutant only modestly affected viral fitness in immortalized cell lines, but reduced viral replication in human airway organoids by 10-fold. In mice, N40D replicated at >1000-fold lower levels compared to the wildtype virus while inducing a robust interferon response; all animals infected with the mutant virus survived infection and showed no signs of lung pathology. Our data validate the SARS-CoV-2 NSP3 Mac1 domain as a critical viral pathogenesis factor and a promising target to develop antivirals.
]]></description>
<dc:creator>Taha, T. Y.</dc:creator>
<dc:creator>Suryawanshi, R. K.</dc:creator>
<dc:creator>Chen, I. P.</dc:creator>
<dc:creator>Correy, G. J.</dc:creator>
<dc:creator>O'Leary, P. C.</dc:creator>
<dc:creator>Jogalekar, M. P.</dc:creator>
<dc:creator>McCavitt-Malvido, M.</dc:creator>
<dc:creator>Diolaiti, M.</dc:creator>
<dc:creator>Kimmerly, G. R.</dc:creator>
<dc:creator>Tsou, C.-L.</dc:creator>
<dc:creator>Martinez-Sobrido, L.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:date>2023-04-18</dc:date>
<dc:identifier>doi:10.1101/2023.04.18.537104</dc:identifier>
<dc:title><![CDATA[A single inactivating amino acid change in the SARS-CoV-2 NSP3 Mac1 domain attenuates viral replication and pathogenesis in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.17.537229v1?rss=1">
<title>
<![CDATA[
In Vitro Modeling of CD8 T Cell Exhaustion Enables CRISPR Screening to Reveal a Role for BHLHE40 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.17.537229v1?rss=1</link>
<description><![CDATA[
Identifying novel molecular mechanisms of exhausted CD8 T cells (Tex) is a key goal of improving immunotherapy of cancer and other diseases. However, high-throughput interrogation of in vivo Tex can be costly and inefficient. In vitro models of Tex are easily customizable and quickly generate high cellular yield, offering an opportunity to perform CRISPR screening and other high-throughput assays. We established an in vitro model of chronic stimulation and benchmarked key phenotypic, functional, transcriptional, and epigenetic features against bona fide in vivo Tex. We leveraged this model of in vitro chronic stimulation in combination with pooled CRISPR screening to uncover transcriptional regulators of T cell exhaustion. This approach identified several transcription factors, including BHLHE40. In vitro and in vivo validation defined a role for BHLHE40 in regulating a key differentiation checkpoint between progenitor and intermediate subsets of Tex. By developing and benchmarking an in vitro model of Tex, we demonstrate the utility of mechanistically annotated in vitro models of Tex, in combination with high-throughput approaches, as a discovery pipeline to uncover novel Tex biology.
]]></description>
<dc:creator>Wu, J. E.</dc:creator>
<dc:creator>Manne, S.</dc:creator>
<dc:creator>Ngiow, S. F.</dc:creator>
<dc:creator>Baxter, A. E.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Freilich, E.</dc:creator>
<dc:creator>Clark, M. L.</dc:creator>
<dc:creator>Lee, J. H.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Khan, O.</dc:creator>
<dc:creator>Staupe, R. P.</dc:creator>
<dc:creator>Huang, Y. J.</dc:creator>
<dc:creator>Shi, J.</dc:creator>
<dc:creator>Giles, J. R.</dc:creator>
<dc:creator>Wherry, E. J.</dc:creator>
<dc:date>2023-04-19</dc:date>
<dc:identifier>doi:10.1101/2023.04.17.537229</dc:identifier>
<dc:title><![CDATA[In Vitro Modeling of CD8 T Cell Exhaustion Enables CRISPR Screening to Reveal a Role for BHLHE40]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.20.536761v1?rss=1">
<title>
<![CDATA[
Conserved enhancer logic controls the notochord expression of vertebrate Brachyury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.20.536761v1?rss=1</link>
<description><![CDATA[
The cell type-specific expression of key transcription factors is central to development. Brachyury/T/TBXT is a major transcription factor for gastrulation, tailbud patterning, and notochord formation; however, how its expression is controlled in the mammalian notochord has remained elusive. Here, we identify the complement of notochord-specific enhancers in the mammalian Brachyury/T/TBXT gene. Using transgenic assays in zebrafish, axolotl, and mouse, we discover three Brachyury-controlling notochord enhancers T3, C, and I in human, mouse, and marsupial genomes. Acting as Brachyury-responsive, auto-regulatory shadow enhancers, deletion of all three enhancers in mouse abolishes Brachyury/T expression selectively in the notochord, causing specific trunk and neural tube defects without gastrulation or tailbud defects. Sequence and functional conservation of Brachyury-driving notochord enhancers with the brachyury/tbxtb loci from diverse lineages of fishes dates their origin to the last common ancestor of jawed vertebrates. Our data define the enhancers for Brachyury/T/TBXTB notochord expression as ancient mechanism in axis development.
]]></description>
<dc:creator>Kemmler, C. L.</dc:creator>
<dc:creator>Smolikova, J.</dc:creator>
<dc:creator>Moran, H. R.</dc:creator>
<dc:creator>Mannion, B. J.</dc:creator>
<dc:creator>Knapp, D.</dc:creator>
<dc:creator>Lim, F.</dc:creator>
<dc:creator>Czarkwiani, A.</dc:creator>
<dc:creator>Hermosilla Aguayo, V.</dc:creator>
<dc:creator>Rapp, V.</dc:creator>
<dc:creator>Fitch, O. E.</dc:creator>
<dc:creator>Boetschi, S.</dc:creator>
<dc:creator>Selleri, L.</dc:creator>
<dc:creator>Farley, E.</dc:creator>
<dc:creator>Braasch, I.</dc:creator>
<dc:creator>Yun, M.</dc:creator>
<dc:creator>Visel, A.</dc:creator>
<dc:creator>Osterwalder, M.</dc:creator>
<dc:creator>Mosimann, C.</dc:creator>
<dc:creator>Kozmik, Z.</dc:creator>
<dc:creator>Burger, A.</dc:creator>
<dc:date>2023-04-20</dc:date>
<dc:identifier>doi:10.1101/2023.04.20.536761</dc:identifier>
<dc:title><![CDATA[Conserved enhancer logic controls the notochord expression of vertebrate Brachyury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.19.537364v1?rss=1">
<title>
<![CDATA[
D-SPIN constructs gene regulatory network models from multiplexed scRNA-seq data revealing organizing principles of cellular perturbation response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.19.537364v1?rss=1</link>
<description><![CDATA[
Gene regulatory networks within cells modulate the expression of the genome in response to signals and changing environmental conditions. Reconstructions of gene regulatory networks can reveal the information processing and control principles used by cells to maintain homeostasis and execute cell-state transitions. Here, we introduce a computational framework, D-SPIN, that generates quantitative models of gene regulatory networks from single-cell mRNA-seq datasets collected across thousands of distinct perturbation conditions. D-SPIN constructs probabilistic models of regulatory interactions between genes or gene-expression programs to fit the cell state distributions under different perturbations. Using large Perturb-seq and drug-response datasets, we demonstrate that D-SPIN models reveal key regulators of cell fate decisions and the coordination of distant cellular pathways in response to gene knockdown perturbations. D-SPIN also dissects gene-level drug response mechanisms in heterogeneous cell populations, elucidating how combinations of immunomodulatory drugs acting on distinct regulators induce novel cell states through additive recruitment of gene expression programs. The D-SPIN model further predicts cell state distributions under drug dosage combinations beyond the training data. D-SPIN provides a computational framework for constructing interpretable models of gene regulatory networks to reveal principles of cellular information processing and physiological control.
]]></description>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Tsou, T.</dc:creator>
<dc:creator>McGinnis, C. S.</dc:creator>
<dc:creator>Khazaei, T.</dc:creator>
<dc:creator>Zhu, Q.</dc:creator>
<dc:creator>Park, J. H.</dc:creator>
<dc:creator>Strazhnik, I.-M.</dc:creator>
<dc:creator>Hanna, J.</dc:creator>
<dc:creator>Chow, E. D.</dc:creator>
<dc:creator>Sivak, D. A.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:date>2023-04-21</dc:date>
<dc:identifier>doi:10.1101/2023.04.19.537364</dc:identifier>
<dc:title><![CDATA[D-SPIN constructs gene regulatory network models from multiplexed scRNA-seq data revealing organizing principles of cellular perturbation response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.19.537546v1?rss=1">
<title>
<![CDATA[
Disease Tolerance Acquired Through Repeated Plasmodium Infection Involves Epigenetic Reprogramming of Innate Immune Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.19.537546v1?rss=1</link>
<description><![CDATA[
The regulation of inflammation is a critical aspect of disease tolerance and naturally acquired immunity to malaria. Here, we demonstrate using RNA sequencing and epigenetic landscape profiling by cytometry by Time-Of-Flight (EpiTOF), that the regulation of inflammatory pathways during asymptomatic parasitemia occurs downstream of pathogen sensing--at the epigenetic level. The abundance of certain epigenetic markers (methylation of H3K27 and dimethylation of arginine residues) and decreased prevalence of histone variant H3.3 correlated with suppressed cytokine responses among monocytes of Ugandan children. Such an epigenetic signature was observed across diverse immune cell populations and not only characterized active asymptomatic parasitemia but also predicted long-term future disease tolerance when observed in uninfected children. This broad methylated signature likely develops gradually and was associated with age and recent parasite exposure. Our data support a model whereby exposure to Plasmodium falciparum induces epigenetic changes that regulate excessive inflammation and contribute to naturally acquired immunity to malaria.
]]></description>
<dc:creator>Nideffer, J.</dc:creator>
<dc:creator>Ty, M.</dc:creator>
<dc:creator>Donato, M.</dc:creator>
<dc:creator>Rek, J.</dc:creator>
<dc:creator>Kajubi, R.</dc:creator>
<dc:creator>Ji, X.</dc:creator>
<dc:creator>Maecker, H.</dc:creator>
<dc:creator>Nankya, F.</dc:creator>
<dc:creator>Musinguzi, K.</dc:creator>
<dc:creator>Press, K.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:creator>Kamya, M.</dc:creator>
<dc:creator>Feeney, M. E.</dc:creator>
<dc:creator>Dorsey, G.</dc:creator>
<dc:creator>Utz, P.</dc:creator>
<dc:creator>Pulendran, B.</dc:creator>
<dc:creator>Khatri, P.</dc:creator>
<dc:creator>Jagannathan, P.</dc:creator>
<dc:date>2023-04-21</dc:date>
<dc:identifier>doi:10.1101/2023.04.19.537546</dc:identifier>
<dc:title><![CDATA[Disease Tolerance Acquired Through Repeated Plasmodium Infection Involves Epigenetic Reprogramming of Innate Immune Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.21.537729v1?rss=1">
<title>
<![CDATA[
Chemical stimuli override a temperature-dependent morphological program by reprogramming the transcriptome of a fungal pathogen. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.21.537729v1?rss=1</link>
<description><![CDATA[
The human fungal pathogen Histoplasma changes its morphology in response to temperature. At 37{degrees}C it grows as a budding yeast whereas at room temperature it transitions to hyphal growth. Prior work has demonstrated that 15-20% of transcripts are temperature-regulated, and that transcription factors Ryp1-4 are necessary to establish yeast growth. However, little is known about transcriptional regulators of the hyphal program. To identify TFs that regulate filamentation, we utilize chemical inducers of hyphal growth. We show that addition of cAMP analogs or an inhibitor of cAMP breakdown overrides yeast morphology, yielding inappropriate hyphal growth at 37{degrees}C. Additionally, butyrate supplementation triggers hyphal growth at 37{degrees}C. Transcriptional profiling of cultures filamenting in response to cAMP or butyrate reveals that a limited set of genes respond to cAMP while butyrate dysregulates a larger set. Comparison of these profiles to previous temperature- or morphology-regulated gene sets identifies a small set of morphology-specific transcripts. This set contains 9 TFs of which we characterized three, STU1, FBC1, and PAC2, whose orthologs regulate development in other fungi. We found that each of these TFs is individually dispensable for room-temperature (RT) induced filamentation but each is required for other aspects of RT development. FBC1 and PAC2, but not STU1, are necessary for filamentation in response to cAMP at 37{degrees}C. Ectopic expression of each of these TFs is sufficient to induce filamentation at 37{degrees}C. Finally, PAC2 induction of filamentation at 37{degrees}C is dependent on STU1, suggesting these TFs form a regulatory circuit that, when activated at RT, promotes the hyphal program.

ImportanceFungal illnesses pose a significant disease burden. However, the regulatory circuits that govern the development and virulence of fungi remain largely unknown. This study utilizes chemicals that can override the normal growth morphology of the human pathogen Histoplasma. Using transcriptomic approaches, we identify novel regulators of hyphal morphology and refine our understanding of the transcriptional circuits governing morphology in Histoplasma.
]]></description>
<dc:creator>Assa, D.</dc:creator>
<dc:creator>Voorhies, M.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:date>2023-04-22</dc:date>
<dc:identifier>doi:10.1101/2023.04.21.537729</dc:identifier>
<dc:title><![CDATA[Chemical stimuli override a temperature-dependent morphological program by reprogramming the transcriptome of a fungal pathogen.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.23.537997v1?rss=1">
<title>
<![CDATA[
Codon affinity in mitochondrial DNA shapes evolutionary and somatic fitness 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.23.537997v1?rss=1</link>
<description><![CDATA[
Summary ParagraphSomatic variation contributes to biological heterogeneity by modulating cellular proclivity to differentiate, expand, adapt, or die. While large-scale sequencing efforts have revealed the foundational role of somatic variants to drive human tumor evolution, our understanding of the contribution of mutations to modulate cellular fitness in non-malignant contexts remains understudied. Here, we identify a mosaic synonymous variant (m.7076A>G) in the mitochondrial DNA (mtDNA) encoded cytochrome c-oxidase subunit 1 gene (MT-CO1, p.Gly391=), which was present at homoplasmy in 47% of immune cells from a healthy donor. Using single-cell multi-omics, we discover highly specific selection against the m.7076G mutant allele in the CD8+ effector memory T cell compartment in vivo, reminiscent of selection observed for pathogenic mtDNA alleles1, 2 and indicative of lineage-specific metabolic requirements. While the wildtype m.7076A allele is translated via Watson-Crick-Franklin base-pairing, the anticodon diversity of the mitochondrial transfer RNA pool is limited, requiring wobble-dependent translation of the m.7076G mutant allele. Notably, mitochondrial ribosome profiling revealed altered codon-anticodon affinity at the wobble position as evidenced by stalled translation of the synonymous m.7076G mutant allele encoding for glycine. Generalizing this observation, we provide a new ontogeny of the 8,482 synonymous variants in the human mitochondrial genome that enables interpretation of functional mtDNA variation. Specifically, via inter- and intra-species evolutionary analyses, population-level complex trait associations, and the occurrence of germline and somatic mtDNA mutations from large-scale sequencing studies, we demonstrate that synonymous variation impacting codon:anticodon affinity is actively evolving across the entire mitochondrial genome and has broad functional and phenotypic effects. In summary, our results introduce a new ontogeny for mitochondrial genetic variation and support a model where organismal principles can be discerned from somatic evolution via single-cell genomics.
]]></description>
<dc:creator>Lareau, C. A.</dc:creator>
<dc:creator>Yin, Y.</dc:creator>
<dc:creator>Gutierrez, J. C.</dc:creator>
<dc:creator>Dhindsa, R. S.</dc:creator>
<dc:creator>Gribling-Burrer, A.-S.</dc:creator>
<dc:creator>Hsieh, Y.-H.</dc:creator>
<dc:creator>Nitsch, L.</dc:creator>
<dc:creator>Buquicchio, F. A.</dc:creator>
<dc:creator>Abay, T.</dc:creator>
<dc:creator>Zielinski, S.</dc:creator>
<dc:creator>Stickels, R. R.</dc:creator>
<dc:creator>Ulirsch, J. C.</dc:creator>
<dc:creator>Yan, P.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Miao, Z.</dc:creator>
<dc:creator>Sandor, K.</dc:creator>
<dc:creator>Daniel, B.</dc:creator>
<dc:creator>Liu, V.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Hu, F.</dc:creator>
<dc:creator>Smith, K. R.</dc:creator>
<dc:creator>Deevi, S. V. V.</dc:creator>
<dc:creator>Maschmeyer, P.</dc:creator>
<dc:creator>Petrovski, S.</dc:creator>
<dc:creator>Smyth, R. P.</dc:creator>
<dc:creator>Greenleaf, W. J.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:creator>Munschauer, M.</dc:creator>
<dc:creator>Ludwig, L. S.</dc:creator>
<dc:creator>Satpathy, A. T.</dc:creator>
<dc:date>2023-04-23</dc:date>
<dc:identifier>doi:10.1101/2023.04.23.537997</dc:identifier>
<dc:title><![CDATA[Codon affinity in mitochondrial DNA shapes evolutionary and somatic fitness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.24.538091v1?rss=1">
<title>
<![CDATA[
PhIP-Seq uncovers novel autoantibodies and unique endotypes in interstitial lung disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.24.538091v1?rss=1</link>
<description><![CDATA[
Interstitial lung diseases (ILDs) are a heterogeneous group of disorders that can develop in patients with connective tissue diseases (CTD). Establishing autoimmunity in ILD impacts prognosis and treatment. ILD patients are screened for autoimmunity by assaying for anti-nuclear autoantibodies, rheumatoid factors and other non-specific tests. However, this approach has not been rigorously validated and may miss autoimmunity that manifests as autoantibodies to tissue antigens not previously defined in ILD. Here, we use Phage Immunoprecipitation-Sequencing (PhIP-Seq) to conduct a large, multi-center unbiased autoantibody discovery screen of ILD patients and controls. PhIP-Seq identified 17 novel autoreactive targets, and machine learning classifiers derived from these targets discriminated ILD serum from controls. Among these 17 candidates, we validated Cadherin Related Family Member 5 (CDHR5) as an autoantigen and found CDHR5 autoantibodies in patients with rheumatologic disorders and importantly, subjects not previously diagnosed with autoimmunity. Lung tissue of CDHR5 autoreactive patients showed transcriptional profiles consistent with activation of NF{kappa}B signaling and upregulation of chitotriosidase (CHIT1), a molecular pathway linked to fibrosis. Our study shows PhIP-Seq uncovers novel autoantibodies in ILD patients not revealed by standard clinical tests. Furthermore, CDHR5 autoantibodies may define a novel molecular endotype of ILD characterized by inflammation and fibrosis.
]]></description>
<dc:creator>Upadhyay, V.</dc:creator>
<dc:creator>Yoon, Y. m.</dc:creator>
<dc:creator>Vazquez, S. E.</dc:creator>
<dc:creator>Velez, T. E.</dc:creator>
<dc:creator>Jones, K. D.</dc:creator>
<dc:creator>Lee, C. T.</dc:creator>
<dc:creator>Law, C. S.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Yang, M. M.</dc:creator>
<dc:creator>Farrand, E.</dc:creator>
<dc:creator>Noth, I.</dc:creator>
<dc:creator>Strek, M. E.</dc:creator>
<dc:creator>Anderson, M.</dc:creator>
<dc:creator>DeRisi, J.</dc:creator>
<dc:creator>Sperling, A. I.</dc:creator>
<dc:creator>Shum, A. K.</dc:creator>
<dc:date>2023-04-24</dc:date>
<dc:identifier>doi:10.1101/2023.04.24.538091</dc:identifier>
<dc:title><![CDATA[PhIP-Seq uncovers novel autoantibodies and unique endotypes in interstitial lung disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.24.537717v1?rss=1">
<title>
<![CDATA[
Transcriptome Analysis Reveals Organ-Specific Effects of 2-Deoxyglucose Treatment in Healthy Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.24.537717v1?rss=1</link>
<description><![CDATA[
ObjectiveGlycolytic inhibition via 2-deoxy-D-glucose (2DG) has potential therapeutic benefits for a range of diseases, including cancer, epilepsy, systemic lupus erythematosus (SLE), and rheumatoid arthritis (RA), and COVID-19, but the systemic effects of 2DG on gene function across different tissues are unclear.

MethodsThis study analyzed the transcriptional profiles of nine tissues from C57BL/6J mice treated with 2DG to understand how it modulates pathways systemically. Principal component analysis (PCA), weighted gene co-network analysis (WGCNA), analysis of variance, and pathway analysis were all performed to identify modules altered by 2DG treatment.

ResultsPCA revealed that samples clustered predominantly by tissue, suggesting that 2DG affects each tissue uniquely. Unsupervised clustering and WGCNA revealed six distinct tissue-specific modules significantly affected by 2DG, each with unique key pathways and genes. 2DG predominantly affected mitochondrial metabolism in the heart, while in the small intestine, it affected immunological pathways.

ConclusionsThese findings suggest that 2DG has a systemic impact that varies across organs, potentially affecting multiple pathways and functions. The study provides insights into the potential therapeutic benefits of 2DG across different diseases and highlights the importance of understanding its systemic effects for future research and clinical applications.
]]></description>
<dc:creator>Wells, A. E.</dc:creator>
<dc:creator>Wilson, J. J.</dc:creator>
<dc:creator>Sears, J. D.</dc:creator>
<dc:creator>Wei, J.</dc:creator>
<dc:creator>Heuer, S.</dc:creator>
<dc:creator>Pandey, R.</dc:creator>
<dc:creator>Costa, M. W.</dc:creator>
<dc:creator>Roopenian, D. C.</dc:creator>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Carter, G. W.</dc:creator>
<dc:date>2023-04-25</dc:date>
<dc:identifier>doi:10.1101/2023.04.24.537717</dc:identifier>
<dc:title><![CDATA[Transcriptome Analysis Reveals Organ-Specific Effects of 2-Deoxyglucose Treatment in Healthy Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.25.538140v1?rss=1">
<title>
<![CDATA[
Tumor cell heterogeneity drives spatial organization of the intratumoral immune response in squamous cell skin carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.25.538140v1?rss=1</link>
<description><![CDATA[
Intratumoral heterogeneity (ITH)--defined as genetic and cellular diversity within a tumor--is linked to failure of immunotherapy and an inferior anti-tumor immune response. The underlying mechanism of this association is unknown. To address this question, we modeled heterogeneous tumors comprised of a pro-inflammatory ("hot") and an immunosuppressive ("cold") tumor population, labeled with YFP and RFP tags respectively to enable precise spatial tracking. The resulting mixed-population tumors exhibited distinct regions comprised of YFP+ (hot) cells, RFP+ (cold) cells, or a mixture. We found that tumor regions occupied by hot tumor cells (YFP+) harbored more total T cells and a higher frequency of Th1 cells and IFN{gamma}+ CD8 T cells compared to regions occupied by cold tumor cells (RFP+), whereas immunosuppressive macrophages showed the opposite spatial pattern. We identified the chemokine CX3CL1, produced at higher levels by our cold tumors, as a mediator of intratumoral macrophage accumulation, particularly immunosuppressive CD206Hi macrophages. Furthermore, we examined the response of heterogeneous tumors to a therapeutic combination of PD-1 blockade and CD40 agonist on a region-by-region basis. While the combination successfully increases Th1 abundance in "cold" tumor regions, it fails to bring overall T cell activity to the same level as seen in "hot" regions. The presence of the "cold" cells thus ultimately leads to a failure of the therapy to induce tumor rejection. Collectively, our results demonstrate that the organization of heterogeneous tumor cells has a profound impact on directing the spatial organization and function of tumor-infiltrating immune cells as well as on responses to immunotherapy.
]]></description>
<dc:creator>Tanaka, M.</dc:creator>
<dc:creator>Lum, L.</dc:creator>
<dc:creator>Hu, K. H.</dc:creator>
<dc:creator>Ledezma-Soto, C.</dc:creator>
<dc:creator>Superville, D. A.</dc:creator>
<dc:creator>Ng, K. M.</dc:creator>
<dc:creator>Adams, Z.</dc:creator>
<dc:creator>Kersten, K. H.</dc:creator>
<dc:creator>Fong, L.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Reeves, M. Q.</dc:creator>
<dc:date>2023-04-26</dc:date>
<dc:identifier>doi:10.1101/2023.04.25.538140</dc:identifier>
<dc:title><![CDATA[Tumor cell heterogeneity drives spatial organization of the intratumoral immune response in squamous cell skin carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.26.538498v1?rss=1">
<title>
<![CDATA[
CRISPRi-based screens in iAssembloids to elucidate neuron-glia interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.26.538498v1?rss=1</link>
<description><![CDATA[
The sheer complexity of the brain has complicated our ability to understand the cellular and molecular mechanisms underlying its function in health and disease. Genome-wide association studies have uncovered genetic variants associated with specific neurological phenotypes and diseases. In addition, single-cell transcriptomics have provided molecular descriptions of specific brain cell types and the changes they undergo during disease. Although these approaches provide a giant leap forward towards understanding how genetic variation can lead to functional changes in the brain, they do not establish molecular mechanisms. To address this need, we developed a 3D co-culture system termed iAssembloids (induced multi-lineage assembloids) that enables the rapid generation of homogenous neuron-glia spheroids. We characterize these iAssembloids with immunohistochemistry and single-cell transcriptomics and combine them with large-scale CRISPRi-based screens. In our first application, we ask how glial and neuronal cells interact to control neuronal death and survival. Our CRISPRi-based screens identified that GSK3{beta} inhibits the protective NRF2-mediated oxidative stress response in the presence of reactive oxygen species elicited by high neuronal activity, which was not previously found in 2D monoculture neuron screens. We also apply the platform to investigate the role of APOE- 4, a risk variant for Alzheimers Disease, in its effect on neuronal survival. We find that APOE- 4-expressing astrocytes may promote more neuronal activity as compared to APOE- 3- expressing astrocytes. This platform expands the toolbox for the unbiased identification of mechanisms of cell-cell interactions in brain health and disease.
]]></description>
<dc:creator>Li, E.</dc:creator>
<dc:creator>Benitez, C.</dc:creator>
<dc:creator>Boggess, S. C.</dc:creator>
<dc:creator>Koontz, M.</dc:creator>
<dc:creator>Rose, I. V. L.</dc:creator>
<dc:creator>Draeger, N.</dc:creator>
<dc:creator>Teter, O. M.</dc:creator>
<dc:creator>Samelson, A. J.</dc:creator>
<dc:creator>Ullian, E. M.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:date>2023-04-27</dc:date>
<dc:identifier>doi:10.1101/2023.04.26.538498</dc:identifier>
<dc:title><![CDATA[CRISPRi-based screens in iAssembloids to elucidate neuron-glia interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.05.539444v1?rss=1">
<title>
<![CDATA[
Up-regulation of cholesterol synthesis pathways and limited neurodegeneration in a knock-in Sod1 mutant mouse model of ALS. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.05.539444v1?rss=1</link>
<description><![CDATA[
Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative disorder affecting brain and spinal cord motor neurons. Mutations in the copper/zinc superoxide dismutase gene (SOD1) are associated with [~]20% of inherited and 1-2% of sporadic ALS cases. Much has been learned from mice expressing transgenic copies of mutant SOD1, which typically involve high-level transgene expression, thereby differing from ALS patients expressing one mutant gene copy. To generate a model that more closely represents patient gene expression, we created a knock-in point mutation (G85R, a human ALS-causing mutation) in the endogenous mouse Sod1 gene, leading to mutant SOD1G85R protein expression. Heterozygous Sod1G85R mutant mice resemble wild type, whereas homozygous mutants have reduced body weight and lifespan, a mild neurodegenerative phenotype, and express very low mutant SOD1 protein levels with no detectable SOD1 activity. Homozygous mutants exhibit partial neuromuscular junction denervation at 3-4 months of age. Spinal cord motor neuron transcriptome analyses of homozygous Sod1G85R mice revealed up-regulation of cholesterol synthesis pathway genes compared to wild type. Transcriptome and phenotypic features of these mice are similar to Sod1 knock-out mice, suggesting the Sod1G85R phenotype is largely driven by loss of SOD1 function. By contrast, cholesterol synthesis genes are down-regulated in severely affected human TgSOD1G93A transgenic mice at 4 months. Our analyses implicate dysregulation of cholesterol or related lipid pathway genes in ALS pathogenesis. The Sod1G85R knock-in mouse is a useful ALS model to examine the importance of SOD1 activity in control of cholesterol homeostasis and motor neuron survival.

SIGNIFICANCE STATEMENTAmyotrophic lateral sclerosis is a devastating disease involving the progressive loss of motor neurons and motor function for which there is currently no cure. Understanding biological mechanisms leading to motor neuron death is critical for developing new treatments. Using a new knock-in mutant mouse model carrying a Sod1 mutation that causes ALS in patients, and in the mouse, causes a limited neurodegenerative phenotype similar to Sod1 loss-of-function, we show that cholesterol synthesis pathway genes are up-regulated in mutant motor neurons, whereas the same genes are down-regulated in transgenic SOD1 mice with a severe phenotype. Our data implicate dysregulation of cholesterol or other related lipid genes in ALS pathogenesis and provide new insights that could contribute to strategies for disease intervention.
]]></description>
<dc:creator>Dominov, J. A.</dc:creator>
<dc:creator>Madigan, L. A.</dc:creator>
<dc:creator>Whitt, J. P.</dc:creator>
<dc:creator>Rademacher, K. L.</dc:creator>
<dc:creator>Webster, K. M.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Banno, H. A.</dc:creator>
<dc:creator>Tang, S.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Wightman, N.</dc:creator>
<dc:creator>Shychuck, E. M.</dc:creator>
<dc:creator>Page, J.</dc:creator>
<dc:creator>Weiss, A.</dc:creator>
<dc:creator>Kelly, K.</dc:creator>
<dc:creator>Kucukural, A. A.</dc:creator>
<dc:creator>Brodsky, M. H.</dc:creator>
<dc:creator>Jaworski, A.</dc:creator>
<dc:creator>Fallon, J. R.</dc:creator>
<dc:creator>Lipscombe, D.</dc:creator>
<dc:creator>Brown, R. H.</dc:creator>
<dc:date>2023-05-05</dc:date>
<dc:identifier>doi:10.1101/2023.05.05.539444</dc:identifier>
<dc:title><![CDATA[Up-regulation of cholesterol synthesis pathways and limited neurodegeneration in a knock-in Sod1 mutant mouse model of ALS.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.05.539601v1?rss=1">
<title>
<![CDATA[
Connectome-based prediction of functional impairment in experimental stroke models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.05.539601v1?rss=1</link>
<description><![CDATA[
Experimental rat models of stroke and hemorrhage are important tools to investigate cerebrovascular disease pathophysi- ology mechanisms, yet how significant patterns of functional impairment induced in various models of stroke are related to changes in connectivity at the level of neuronal populations and mesoscopic parcellations of rat brains remain unresolved. To address this gap in knowledge, we employed two middle cerebral artery occlusion models and one intracerebral hemorrhage model with variant extent and location of neuronal dysfunction. Motor and spatial memory function was assessed and the level of hippocampal activation via Fos immunohistochemistry. Contribution of connectivity change to functional impairment was analyzed for connection similarities, graph distances and spatial distances as well as the importance of regions in terms of network architecture based on the neuroVIISAS rat connectome. We found that functional impairment correlated with not only the extent but also the locations of the injury among the models. In addition, via coactivation analysis in dynamic rat brain models, we found that lesioned regions led to stronger coactivations with motor function and spatial learning regions than with other unaffected regions of the connectome. Dynamic modeling with the weighted bilateral connectome detected changes in signal propagation in the remote hippocampus in all 3 stroke types, predicting the extent of hippocampal hypoactivation and impairment in spatial learning and memory function. Our study provides a comprehensive analytical framework in predictive identification of remote regions not directly altered by stroke events and their functional implication.
]]></description>
<dc:creator>Schmitt, O.</dc:creator>
<dc:creator>Eipert, P.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Kanoke, A.</dc:creator>
<dc:creator>Rabiller, G.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:date>2023-05-05</dc:date>
<dc:identifier>doi:10.1101/2023.05.05.539601</dc:identifier>
<dc:title><![CDATA[Connectome-based prediction of functional impairment in experimental stroke models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.08.539503v1?rss=1">
<title>
<![CDATA[
Anatomical registration of intracranial electrodes. Robust model-based localization and deformable smooth brain-shift compensation methods 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.08.539503v1?rss=1</link>
<description><![CDATA[
Precise electrode localization is important for maximizing the utility of intracranial EEG data. Electrodes are typically localized from post-implantation CT artifacts, but algorithms can fail due to low signal-to-noise ratio, unrelated artifacts, or high-density electrode arrays. Minimizing these errors usually requires time-consuming visual localization and can still result in inaccurate localizations. In addition, surgical implantation of grids and strips typically introduces non-linear brain deformations, which result in anatomical registration errors when post-implantation CT images are fused with the pre-implantation MRI images. Several projection methods are currently available, but they either fail to produce smooth solutions or do not account for brain deformations.

To address these shortcomings, we propose two novel algorithms for the anatomical registration of intracranial electrodes that are almost fully automatic and provide highly accurate results. We first present GridFit, an algorithm that simultaneously localizes all contacts in grids, strips, or depth arrays by fitting flexible models to the electrodes CT artifacts. We observed localization errors of less than one millimeter (below 8% relative to the inter-electrode distance) and robust performance under the presence of noise, unrelated artifacts, and high-density implants when we ran [~]6000 simulated scenarios. Furthermore, we validated the method with real data from 20 intracranial patients.

As a second registration step, we introduce CEPA, a brain-shift compensation algorithm that combines orthogonal-based projections, spring-mesh models, and spatial regularization constraints. When tested with real data from 15 patients, anatomical registration errors were smaller than those obtained for well-established alternatives. Additionally, CEPA accounted simultaneously for simple mechanical deformation principles, which is not possible with other available methods. Inter-electrode distances of projected coordinates smoothly changed across neighbor electrodes, while changes in inter-electrode distances linearly increased with projection distance. Moreover, in an additional validation procedure, we found that modeling resting-state high-frequency activity (75-145 Hz) in five patients further supported our new algorithm.

Together, GridFit and CEPA constitute a versatile set of tools for the registration of subdural grid, strip, and depth electrode coordinates that provide highly accurate results even in the most challenging implantation scenarios. The methods presented here are implemented in the iElectrodes open-source toolbox, making their use simple, accessible, and straightforward to integrate with other popular toolboxes used for analyzing electrophysiological data.
]]></description>
<dc:creator>Blenkmann, A. O.</dc:creator>
<dc:creator>Leske, S. L.</dc:creator>
<dc:creator>Llorens, A.</dc:creator>
<dc:creator>Lin, J. J.</dc:creator>
<dc:creator>Chang, E.</dc:creator>
<dc:creator>Brunner, P.</dc:creator>
<dc:creator>Schalk, G.</dc:creator>
<dc:creator>Ivanovic, J.</dc:creator>
<dc:creator>Larsson, P. G.</dc:creator>
<dc:creator>Knight, R. T.</dc:creator>
<dc:creator>Endestad, T.</dc:creator>
<dc:creator>Solbakk, A.-K.</dc:creator>
<dc:date>2023-05-09</dc:date>
<dc:identifier>doi:10.1101/2023.05.08.539503</dc:identifier>
<dc:title><![CDATA[Anatomical registration of intracranial electrodes. Robust model-based localization and deformable smooth brain-shift compensation methods]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.09.539589v1?rss=1">
<title>
<![CDATA[
The developmental timing of spinal touch processing alterations and its relation to ASD-associated behaviors in mouse models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.09.539589v1?rss=1</link>
<description><![CDATA[
Altered somatosensory reactivity is frequently observed among individuals with autism spectrum disorders (ASDs). Here, we report that while multiple mouse models of ASD exhibit aberrant somatosensory behaviors in adulthood, some models exhibit altered tactile reactivity as early as embryonic development, while in others, altered reactivity emerges later in life. Additionally, tactile over-reactivity during neonatal development is associated with anxiety-like behaviors and social interaction deficits in adulthood, whereas tactile over-reactivity that emerges later in life is not. The locus of circuit disruption dictates the timing of aberrant tactile behaviors: altered feedback or presynaptic inhibition of peripheral mechanosensory neurons leads to abnormal tactile reactivity during neonatal development, while disruptions in feedforward inhibition in the spinal cord lead to touch reactivity alterations that manifest later in life. Thus, the developmental timing of aberrant touch processing can predict the manifestation of ASD-associated behaviors in mouse models, and differential timing of sensory disturbance onset may contribute to phenotypic diversity across individuals with ASD.
]]></description>
<dc:creator>Tasnim, A.</dc:creator>
<dc:creator>Alkislar, I.</dc:creator>
<dc:creator>Hakim, R.</dc:creator>
<dc:creator>Turecek, J.</dc:creator>
<dc:creator>Abdelaziz, A.</dc:creator>
<dc:creator>Orefice, L. L.</dc:creator>
<dc:creator>Ginty, D. D.</dc:creator>
<dc:date>2023-05-09</dc:date>
<dc:identifier>doi:10.1101/2023.05.09.539589</dc:identifier>
<dc:title><![CDATA[The developmental timing of spinal touch processing alterations and its relation to ASD-associated behaviors in mouse models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.08.539863v1?rss=1">
<title>
<![CDATA[
Gene networks reveal stem-cell state convergence during preneoplasia and progression to malignancy in multistage skin carcinogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.08.539863v1?rss=1</link>
<description><![CDATA[
Adult mammalian stem cells play critical roles in normal tissue homeostasis, as well as in tumor development, by contributing to cell heterogeneity, plasticity, and development of drug resistance. The relationship between different types of normal and cancer stem cells is highly controversial and poorly understood. Here, we carried out gene expression network analysis of normal and tumor samples from genetically heterogeneous mice to create network metagenes for visualization of stem-cell networks, rather than individual stem-cell markers, at the single-cell level during multistage carcinogenesis. We combined this approach with lineage tracing and single-cell RNASeq of stem cells and their progeny, identifying a previously unrecognized hierarchy in which Lgr6+ stem cells from tumors generate progeny that express a range of other stem-cell markers including Sox2, Pitx1, Foxa1, Klf5, and Cd44. Our data identify a convergence of multiple stem-cell and tumor-suppressor pathways in benign tumor cells expressing markers of lineage plasticity and oxidative stress. This same single-cell population expresses network metagenes corresponding to markers of cancer drug resistance in human tumors of the skin, lung and prostate. Treatment of mouse squamous carcinomas in vivo with the chemotherapeutic cis-platin resulted in elevated expression of the genes that mark this cell population. Our data have allowed us to create a simplified model of multistage carcinogenesis that identifies distinct stem-cell states at different stages of tumor progression, thereby identifying networks involved in lineage plasticity, drug resistance, and immune surveillance, providing a rich source of potential targets for cancer therapy.

One-Sentence SummaryGenes act in networks to drive cancer, and we identify these groups of genes from bulk-tissue and trace them at single-cell resolution.
]]></description>
<dc:creator>Taylor, M. A.</dc:creator>
<dc:creator>Kandyba, E.</dc:creator>
<dc:creator>Halliwell, K.</dc:creator>
<dc:creator>Del Rosario, R.</dc:creator>
<dc:creator>Khoroshkin, M. S.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Quigley, D.</dc:creator>
<dc:creator>Li, Y. R.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Bollam, S.</dc:creator>
<dc:creator>Mirzoeva, O.</dc:creator>
<dc:creator>Akhurst, R.</dc:creator>
<dc:creator>Balmain, A.</dc:creator>
<dc:date>2023-05-10</dc:date>
<dc:identifier>doi:10.1101/2023.05.08.539863</dc:identifier>
<dc:title><![CDATA[Gene networks reveal stem-cell state convergence during preneoplasia and progression to malignancy in multistage skin carcinogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.09.539936v1?rss=1">
<title>
<![CDATA[
Anatomical and behavioral correlates of auditory perception in developmental dyslexia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.09.539936v1?rss=1</link>
<description><![CDATA[
Developmental dyslexia is typically associated with difficulties in basic auditory processing and in manipulating speech sounds. However, the neuroanatomical correlates of auditory difficulties in developmental dyslexia (DD) and their contribution to individual clinical phenotypes are still unknown. Recent intracranial electrocorticography findings associated processing of sound amplitude rises and speech sounds with posterior and middle superior temporal gyrus (STG), respectively. We hypothesize that regional STG anatomy will relate to specific auditory abilities in DD, and that auditory processing abilities will relate to behavioral difficulties with speech and reading. One hundred and ten children (78 DD, 32 typically developing, age 7-15 years) completed amplitude rise time and speech in noise discrimination tasks. They also underwent a battery of cognitive tests. Anatomical MRI scans were used to identify regions in which local cortical gyrification complexity correlated with auditory behavior. Behaviorally, amplitude rise time but not speech in noise performance was impaired in DD. Neurally, amplitude rise time and speech in noise performance correlated with gyrification in posterior and middle STG, respectively. Furthermore, amplitude rise time significantly contributed to reading impairments in DD, while speech in noise only explained variance in phonological awareness. Finally, amplitude rise time and speech in noise performance were not correlated, and each task was correlated with distinct neuropsychological measures, emphasizing their unique contributions to DD. Overall, we provide a direct link between the neurodevelopment of the left STG and individual variability in auditory processing abilities in neurotypical and dyslexic populations.
]]></description>
<dc:creator>Qi, T.</dc:creator>
<dc:creator>Mandelli, M. L.</dc:creator>
<dc:creator>Watson Pereira, C. L.</dc:creator>
<dc:creator>Wellman, E.</dc:creator>
<dc:creator>Bogley, R.</dc:creator>
<dc:creator>Licata, A. E.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Oganian, Y.</dc:creator>
<dc:creator>Gorno-Tempini, M. L.</dc:creator>
<dc:date>2023-05-10</dc:date>
<dc:identifier>doi:10.1101/2023.05.09.539936</dc:identifier>
<dc:title><![CDATA[Anatomical and behavioral correlates of auditory perception in developmental dyslexia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.11.540281v1?rss=1">
<title>
<![CDATA[
Allosteric regulation of TRPV1 by the inflammatory lipid, LPA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540281v1?rss=1</link>
<description><![CDATA[
TRP ion channels are modulated by phosphoinositide lipids, but the underlying structural mechanisms remain unclear. The capsaicin- and heat-activated receptor, TRPV1, has served as a model for deciphering lipid modulation, which is relevant to understanding how pro-algesic agents enhance channel activity in the setting of inflammatory pain. Identification of a pocket within the TRPV1 transmembrane core has provided initial clues as to how phosphoinositide lipids bind to and regulate the channel. Here we show that this regulatory pocket can accommodate diverse lipid species, including the inflammatory lipid lysophosphatidic acid (LPA), whose actions are determined by their specific modes of binding. Furthermore, we show that an  empty pocket channel lacking an endogenous phosphoinositide lipid assumes an agonist-like state, even at low temperature, substantiating the concept that phosphoinositide lipids serve as negative TRPV1 modulators whose ejection from the binding pocket is a critical step towards activation by thermal or chemical stimuli.
]]></description>
<dc:creator>Arnold, W. R.</dc:creator>
<dc:creator>Julius, D.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:date>2023-05-11</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540281</dc:identifier>
<dc:title><![CDATA[Allosteric regulation of TRPV1 by the inflammatory lipid, LPA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.11.540338v1?rss=1">
<title>
<![CDATA[
Circadian PERIOD complex regulates TC-DSB repair through anchoring to the nuclear envelope 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540338v1?rss=1</link>
<description><![CDATA[
Repair of DNA Double-Strand Breaks (DSBs) produced in transcriptionally active chromatin occurs through a poorly characterized pathway called Transcription-Coupled DSB repair (TC-DSBR). Here, using a screening approach scoring multiple outputs in human cells, we identified proteins from the PERIOD complex, a key module ensuring circadian oscillations, as novel TC-DSBR players. We show that the core PER complex protein PER2 is recruited at TC-DSBs and that it contributes to the targeting of TC-DSBs at the nuclear envelope (NE). At the NE, SUN1 and the Nuclear Pore Complex (NPC) act as docking sites for TC-DSBs and TC-DSB anchoring fosters RAD51 assembly. Impaired DSB localization at the NE results in elevated DSB clustering and translocation rate. In agreement, the circadian clock regulates TC-DSB anchoring to the NE, RAD51 assembly, and DSB clustering. Our study shows that DSB localization to the NPC is a conserved molecular pathway that also occurs in human cells and provides a direct link between the circadian rhythm and the response to DSBs occurring in active genes. This opens new therapeutic strategies for chemotherapies based on drugs that are inducing DSBs in active loci such as topoisomerase poisons.
]]></description>
<dc:creator>Le Bozec, B.</dc:creator>
<dc:creator>Guitton-Sert, L.</dc:creator>
<dc:creator>Collins, S.</dc:creator>
<dc:creator>Rocher, V.</dc:creator>
<dc:creator>Guillou, E.</dc:creator>
<dc:creator>Payrault, C.</dc:creator>
<dc:creator>Arnould, C.</dc:creator>
<dc:creator>Guenole, A.</dc:creator>
<dc:creator>Aguirrebengoa, M.</dc:creator>
<dc:creator>Finoux, A.-L.</dc:creator>
<dc:creator>Mangeat, T.</dc:creator>
<dc:creator>Puget, N.</dc:creator>
<dc:creator>Legube, G.</dc:creator>
<dc:date>2023-05-11</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540338</dc:identifier>
<dc:title><![CDATA[Circadian PERIOD complex regulates TC-DSB repair through anchoring to the nuclear envelope]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.10.539944v1?rss=1">
<title>
<![CDATA[
Respiratory Complex I Regulates Dendritic Cell Maturation in Explant Model of Human Tumor Immune Microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.10.539944v1?rss=1</link>
<description><![CDATA[
Combining cytotoxic chemotherapy or novel anticancer drugs with T-cell modulators holds great promise in treating advanced cancers. However, the response varies depending on the tumor immune microenvironment (TIME). Therefore, there is a clear need for pharmacologically tractable models of the TIME to dissect its influence on mono- and combination treatment response at the individual level. Here we establish a Patient-Derived Explant Culture (PDEC) model of breast cancer, which retains the immune contexture of the primary tumor, recapitulating cytokine profiles and CD8+ T cell cytotoxic activity. We explored the immunomodulatory action of a synthetic lethal BCL2 inhibitor venetoclax + metformin drug combination ex vivo, discovering metformin cannot overcome the lymphocyte-depleting action of venetoclax. Instead, metformin promotes dendritic cell maturation through inhibition of mitochondrial complex I, increasing their capacity to co-stimulate CD4+ T cells and thus facilitating anti-tumor immunity. Our results establish PDECs as a feasible model to identify immunomodulatory functions of anticancer drugs in the context of patient-specific TIME.
]]></description>
<dc:creator>Turpin, R.</dc:creator>
<dc:creator>Liu, R.</dc:creator>
<dc:creator>Munne, P.</dc:creator>
<dc:creator>Peura, A.</dc:creator>
<dc:creator>Rannikko, J. H.</dc:creator>
<dc:creator>Philips, G.</dc:creator>
<dc:creator>Boeckx, B.</dc:creator>
<dc:creator>Salmelin, N.</dc:creator>
<dc:creator>Hurskainen, E.</dc:creator>
<dc:creator>Suleymanova, I.</dc:creator>
<dc:creator>Vuorinen, E. M.</dc:creator>
<dc:creator>Lehtinen, L.</dc:creator>
<dc:creator>Mutka, M.</dc:creator>
<dc:creator>Kovanen, P. E.</dc:creator>
<dc:creator>Niinikoski, L.</dc:creator>
<dc:creator>Meretoja, T.</dc:creator>
<dc:creator>Mattson, J.</dc:creator>
<dc:creator>Mustjoki, S.</dc:creator>
<dc:creator>Saavalainen, P.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:creator>Lambrechts, D.</dc:creator>
<dc:creator>Pouwels, J.</dc:creator>
<dc:creator>Hollmen, M.</dc:creator>
<dc:creator>Klefstrom, J.</dc:creator>
<dc:date>2023-05-12</dc:date>
<dc:identifier>doi:10.1101/2023.05.10.539944</dc:identifier>
<dc:title><![CDATA[Respiratory Complex I Regulates Dendritic Cell Maturation in Explant Model of Human Tumor Immune Microenvironment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.11.540322v1?rss=1">
<title>
<![CDATA[
A ubiquitous GC content signature underlies multimodal mRNA regulation by DDX3X 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540322v1?rss=1</link>
<description><![CDATA[
The road from transcription to protein synthesis is paved with many obstacles, allowing for several modes of post-transcriptional regulation of gene expression. A fundamental player in mRNA biology is DDX3X, an RNA binding protein that canonically regulates mRNA translation. By monitoring dynamics of mRNA abundance and translation following DDX3X depletion, we observe stabilization of translationally suppressed mRNAs. We use interpretable statistical learning models to uncover GC content in the coding sequence as the major feature underlying RNA stabilization. This result corroborates GC content-related mRNA regulation detectable in other studies, including hundreds of ENCODE datasets and recent work focusing on mRNA dynamics in the cell cycle. We provide further evidence for mRNA stabilization by detailed analysis of RNA-seq profiles in hundreds of samples, including a Ddx3x conditional knockout mouse model exhibiting cell cycle and neurogenesis defects. Our study identifies a ubiquitous feature underlying mRNA regulation and highlights the importance of quantifying multiple steps of the gene expression cascade, where RNA abundance and protein production are often uncoupled.
]]></description>
<dc:creator>Jowhar, Z.</dc:creator>
<dc:creator>Xu, A.</dc:creator>
<dc:creator>Venkataramanan, S.</dc:creator>
<dc:creator>Hoye, M. L.</dc:creator>
<dc:creator>Silver, D. L.</dc:creator>
<dc:creator>Floor, S. N.</dc:creator>
<dc:creator>Calviello, L.</dc:creator>
<dc:date>2023-05-12</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540322</dc:identifier>
<dc:title><![CDATA[A ubiquitous GC content signature underlies multimodal mRNA regulation by DDX3X]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.11.540409v1?rss=1">
<title>
<![CDATA[
Time-resolved network control analysis links reduced control energy under DMT with the serotonin 2a receptor, signal diversity, and subjective experience. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540409v1?rss=1</link>
<description><![CDATA[
Psychedelics offer a profound window into the functioning of the human brain and mind through their robust acute effects on perception, subjective experience, and brain activity patterns. In recent work using a receptor-informed network control theory framework, we demonstrated that the serotonergic psychedelics lysergic acid diethylamide (LSD) and psilocybin flatten the brains control energy landscape in a manner that covaries with more dynamic and entropic brain activity. Contrary to LSD and psilocybin, whose effects last for hours, the serotonergic psychedelic N,N-dimethyltryptamine (DMT) rapidly induces a profoundly immersive altered state of consciousness lasting less than 20 minutes, allowing for the entirety of the drug experience to be captured during a single resting-state fMRI scan. Using network control theory, which quantifies the amount of input necessary to drive transitions between functional brain states, we integrate brain structure and function to map the energy trajectories of 14 individuals undergoing fMRI during DMT and placebo. Consistent with previous work, we find that global control energy is reduced following injection with DMT compared to placebo. We additionally show longitudinal trajectories of global control energy correlate with longitudinal trajectories of EEG signal diversity (a measure of entropy) and subjective ratings of drug intensity. We interrogate these same relationships on a regional level and find that the spatial patterns of DMTs effects on these metrics are correlated with serotonin 2a receptor density (obtained from separately acquired PET data). Using receptor distribution and pharmacokinetic information, we were able to successfully recapitulate the effects of DMT on global control energy trajectories, demonstrating a proof-of-concept for the use of control models in predicting pharmacological intervention effects on brain dynamics.
]]></description>
<dc:creator>Singleton, S. P.</dc:creator>
<dc:creator>Timmermann, C.</dc:creator>
<dc:creator>Luppi, A. I.</dc:creator>
<dc:creator>Eckernas, E.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Carhart-Harris, R. L.</dc:creator>
<dc:creator>Kuceyeski, A.</dc:creator>
<dc:date>2023-05-12</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540409</dc:identifier>
<dc:title><![CDATA[Time-resolved network control analysis links reduced control energy under DMT with the serotonin 2a receptor, signal diversity, and subjective experience.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.12.540515v1?rss=1">
<title>
<![CDATA[
Quantifying the relationship between cell proliferation and morphology during development of the face 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.12.540515v1?rss=1</link>
<description><![CDATA[
Morphogenesis requires highly coordinated, complex interactions between cellular processes: proliferation, migration, and apoptosis, along with physical tissue interactions. How these cellular and tissue dynamics drive morphogenesis remains elusive. Three dimensional (3D) microscopic imaging poses great promise, and generates elegant images. However, generating even moderate through-put quantified images is challenging for many reasons. As a result, the association between morphogenesis and cellular processes in 3D developing tissues has not been fully explored. To address this critical gap, we have developed an imaging and image analysis pipeline to enable 3D quantification of cellular dynamics along with 3D morphology for the same individual embryo. Specifically, we focus on how 3D distribution of proliferation relates to morphogenesis during mouse facial development. Our method involves imaging with light-sheet microscopy, automated segmentation of cells and tissues using machine learning-based tools, and quantification of external morphology via geometric morphometrics. Applying this framework, we show that changes in proliferation are tightly correlated to changes in morphology over the course of facial morphogenesis. These analyses illustrate the potential of this pipeline to investigate mechanistic relationships between cellular dynamics and morphogenesis during embryonic development.
]]></description>
<dc:creator>Green, R. M.</dc:creator>
<dc:creator>Lo Vercio, L.</dc:creator>
<dc:creator>Dauter, A.</dc:creator>
<dc:creator>Barretto, E. C.</dc:creator>
<dc:creator>Devine, J.</dc:creator>
<dc:creator>Vidal Garcia, M.</dc:creator>
<dc:creator>Marchini, M.</dc:creator>
<dc:creator>Robertson, S.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Mahika, A.</dc:creator>
<dc:creator>Shakir, M. B.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:creator>Boughner, J. C.</dc:creator>
<dc:creator>Dean, W.</dc:creator>
<dc:creator>Lander, A. D.</dc:creator>
<dc:creator>Marcucio, R. S.</dc:creator>
<dc:creator>Forkert, N. D.</dc:creator>
<dc:creator>Hallgrimsson, B.</dc:creator>
<dc:date>2023-05-12</dc:date>
<dc:identifier>doi:10.1101/2023.05.12.540515</dc:identifier>
<dc:title><![CDATA[Quantifying the relationship between cell proliferation and morphology during development of the face]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.15.540864v1?rss=1">
<title>
<![CDATA[
The p97/VCP adapter UBXD1 drives AAA+ remodeling and ring opening through multi-domain tethered interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.15.540864v1?rss=1</link>
<description><![CDATA[
p97/VCP is an essential cytosolic AAA+ ATPase hexamer that extracts and unfolds substrate polypeptides during protein homeostasis and degradation. Distinct sets of p97 adapters guide cellular functions but their roles in direct control of the hexamer are unclear. The UBXD1 adapter localizes with p97 in critical mitochondria and lysosome clearance pathways and contains multiple p97-interacting domains. We identify UBXD1 as a potent p97 ATPase inhibitor and report structures of intact p97:UBXD1 complexes that reveal extensive UBXD1 contacts across p97 and an asymmetric remodeling of the hexamer. Conserved VIM, UBX, and PUB domains tether adjacent protomers while a connecting strand forms an N-terminal domain lariat with a helix wedged at the interprotomer interface. An additional VIM-connecting helix binds along the second AAA+ domain. Together these contacts split the hexamer into a ring-open conformation. Structures, mutagenesis, and comparisons to other adapters further reveal how adapters containing conserved p97-remodeling motifs regulate p97 ATPase activity and structure.
]]></description>
<dc:creator>Braxton, J. R.</dc:creator>
<dc:creator>Altobelli, C. R.</dc:creator>
<dc:creator>Tucker, M. R.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Thwin, A. C.</dc:creator>
<dc:creator>Arkin, M. R.</dc:creator>
<dc:creator>Southworth, D. R.</dc:creator>
<dc:date>2023-05-15</dc:date>
<dc:identifier>doi:10.1101/2023.05.15.540864</dc:identifier>
<dc:title><![CDATA[The p97/VCP adapter UBXD1 drives AAA+ remodeling and ring opening through multi-domain tethered interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.15.540872v1?rss=1">
<title>
<![CDATA[
Asymmetric apical domain states of mitochondrial Hsp60 coordinate substrate engagement and chaperonin assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.15.540872v1?rss=1</link>
<description><![CDATA[
The mitochondrial chaperonin, mtHsp60, promotes the folding of newly imported and transiently misfolded proteins in the mitochondrial matrix, assisted by its co-chaperone mtHsp10. Despite its essential role in mitochondrial proteostasis, structural insights into how this chaperonin binds to clients and progresses through its ATP-dependent reaction cycle are not clear. Here, we determined cryo-electron microscopy (cryo-EM) structures of a hyperstable disease-associated mtHsp60 mutant, V72I, at three stages in this cycle. Unexpectedly, client density is identified in all states, revealing interactions with mtHsp60s apical domains and C-termini that coordinate client positioning in the folding chamber. We further identify a striking asymmetric arrangement of the apical domains in the ATP state, in which an alternating up/down configuration positions interaction surfaces for simultaneous recruitment of mtHsp10 and client retention. Client is then fully encapsulated in mtHsp60/mtHsp10, revealing prominent contacts at two discrete sites that potentially support maturation. These results identify a new role for the apical domains in coordinating client capture and progression through the cycle, and suggest a conserved mechanism of group I chaperonin function.
]]></description>
<dc:creator>Braxton, J. R.</dc:creator>
<dc:creator>Shao, H.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Southworth, D. R.</dc:creator>
<dc:date>2023-05-15</dc:date>
<dc:identifier>doi:10.1101/2023.05.15.540872</dc:identifier>
<dc:title><![CDATA[Asymmetric apical domain states of mitochondrial Hsp60 coordinate substrate engagement and chaperonin assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.15.540704v1?rss=1">
<title>
<![CDATA[
Neutrophils actively swell to potentiate rapid migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.15.540704v1?rss=1</link>
<description><![CDATA[
While the involvement of actin polymerization in cell migration is well-established, much less is known about the role of transmembrane water flow in cell motility. Here, we investigate the role of water influx in a prototypical migrating cell, the neutrophil, which undergoes rapid, directed movement to sites of injury and infection. Chemoattractant exposure both increases cell volume and potentiates migration, but the causal link between these processes is not known. We combine single cell volume measurements and a genome-wide CRISPR screen to identify the regulators of chemoattractant-induced neutrophil swelling, including NHE1, AE2, PI3K-gamma, and CA2. Through NHE1 inhibition in primary human neutrophils, we show that cell swelling is both necessary and sufficient for the potentiation of migration following chemoattractant stimulation. Our data demonstrate that chemoattractant-driven cell swelling complements cytoskeletal rearrangements to enhance migration speed.
]]></description>
<dc:creator>Nagy, T. L.</dc:creator>
<dc:creator>Strickland, J. D.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:date>2023-05-16</dc:date>
<dc:identifier>doi:10.1101/2023.05.15.540704</dc:identifier>
<dc:title><![CDATA[Neutrophils actively swell to potentiate rapid migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.15.540305v1?rss=1">
<title>
<![CDATA[
Multiomic Single Cell Sequencing Identifies Stemlike Nature of Mixed Phenotype Acute Leukemia and Provides Novel Risk Stratification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.15.540305v1?rss=1</link>
<description><![CDATA[
Mixed phenotype acute leukemia (MPAL) is a leukemia whose biologic drivers are poorly understood, therapeutic strategy remains unclear, and prognosis is poor. We performed multiomic single cell (SC) profiling of 14 newly diagnosed adult MPAL patients to characterize the immunophenotypic, genetic, and transcriptional landscapes of MPAL. We show that neither genetic profile nor transcriptome reliably correlate with specific MPAL immunophenotypes. However, progressive acquisition of mutations is associated with increased expression of immunophenotypic markers of immaturity. Using SC transcriptional profiling, we find that MPAL blasts express a stem cell-like transcriptional profile distinct from other acute leukemias and indicative of high differentiation potential. Further, patients with the highest differentiation potential demonstrated inferior survival in our dataset. A gene set score, MPAL95, derived from genes highly enriched in this cohort, is applicable to bulk RNA sequencing data and was predictive of survival in an independent patient cohort, suggesting utility for clinical risk stratification.
]]></description>
<dc:creator>Peretz, C. A. C.</dc:creator>
<dc:creator>Kennedy, V. E.</dc:creator>
<dc:creator>Walia, A.</dc:creator>
<dc:creator>Delley, C. L.</dc:creator>
<dc:creator>Koh, A.</dc:creator>
<dc:creator>Tran, E.</dc:creator>
<dc:creator>Clark, I.</dc:creator>
<dc:creator>Hayford, C. E.</dc:creator>
<dc:creator>D'Amato, C.</dc:creator>
<dc:creator>Xue, Y.</dc:creator>
<dc:creator>Fontanez, K. M.</dc:creator>
<dc:creator>Roy, R.</dc:creator>
<dc:creator>Logan, A. C.</dc:creator>
<dc:creator>Perl, A. E.</dc:creator>
<dc:creator>Abate, A.</dc:creator>
<dc:creator>Olshen, A.</dc:creator>
<dc:creator>Smith, C. C.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.15.540305</dc:identifier>
<dc:title><![CDATA[Multiomic Single Cell Sequencing Identifies Stemlike Nature of Mixed Phenotype Acute Leukemia and Provides Novel Risk Stratification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.15.540834v1?rss=1">
<title>
<![CDATA[
Multivalent coiled-coil interactions enable full-scale centrosome assembly and strength 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.15.540834v1?rss=1</link>
<description><![CDATA[
During mitotic spindle assembly, microtubules generate tensile stresses on pericentriolar material (PCM), the outermost layer of centrosomes. The molecular interactions that enable PCM to assemble rapidly and resist external forces are unknown. Here we use cross-linking mass spectrometry to identify interactions underlying supramolecular assembly of SPD-5, the main PCM scaffold protein in C. elegans. Crosslinks map primarily to alpha helices within the phospho-regulated region (PReM), a long C-terminal coiled-coil, and a series of four N-terminal coiled-coils. PLK-1 phosphorylation of SPD-5 creates new homotypic contacts, including two between PReM and the CM2-like domain, and eliminates numerous contacts in disordered linker regions, thus favoring coiled-coil-specific interactions. Mutations within these interacting regions cause PCM assembly defects that are partly rescued by eliminating microtubule-mediated forces. Thus, PCM assembly and strength are interdependent. In vitro, self-assembly of SPD-5 scales with coiled-coil content, although there is a defined hierarchy of association. We propose that multivalent interactions among coiled-coil regions of SPD-5 build the PCM scaffold and contribute sufficient strength to resist microtubule-mediated forces.
]]></description>
<dc:creator>Rios, M. U.</dc:creator>
<dc:creator>Bagnucka, M. A.</dc:creator>
<dc:creator>Ryder, B. D.</dc:creator>
<dc:creator>Gomes, B. F.</dc:creator>
<dc:creator>Familiari, N.</dc:creator>
<dc:creator>Yaguchi, K.</dc:creator>
<dc:creator>Amato, M.</dc:creator>
<dc:creator>Joachimiak, Łukasz A.</dc:creator>
<dc:creator>Woodruff, J. B.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.15.540834</dc:identifier>
<dc:title><![CDATA[Multivalent coiled-coil interactions enable full-scale centrosome assembly and strength]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.16.540868v1?rss=1">
<title>
<![CDATA[
A central helical hairpin in SPD-5 enables centrosome strength and assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.16.540868v1?rss=1</link>
<description><![CDATA[
Centrosomes organize microtubules for mitotic spindle assembly and positioning. Forces mediated by these microtubules create tensile stresses on pericentriolar material (PCM), the outermost layer of centrosomes. How PCM resists these stresses is unclear at the molecular level. Here, we use cross-linking mass spectrometry (XL-MS) to map interactions underlying multimerization of SPD-5, an essential PCM scaffold component in C. elegans. We identified an interaction hotspot in an alpha helical hairpin motif in SPD-5 (a.a. 541-677). XL-MS data, ab initio structural predictions, and mass photometry suggest that this region dimerizes to form a tetrameric coiled-coil. Mutating a helical section (a.a. 610-640) or a single residue (R592) inhibited PCM assembly in embryos. This phenotype was rescued by eliminating microtubule pulling forces, revealing that PCM assembly and material strength are interrelated. We propose that interactions mediated by the helical hairpin strongly bond SPD-5 molecules to each other, thus enabling PCM to assemble fully and withstand stresses generated by microtubules.
]]></description>
<dc:creator>Rios, M. U.</dc:creator>
<dc:creator>Ryder, B. D.</dc:creator>
<dc:creator>Familiari, N.</dc:creator>
<dc:creator>Joachimiak, Łukasz A.</dc:creator>
<dc:creator>Woodruff, J. B.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.16.540868</dc:identifier>
<dc:title><![CDATA[A central helical hairpin in SPD-5 enables centrosome strength and assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.17.541057v1?rss=1">
<title>
<![CDATA[
Recovery of a learned behavior despite partial restoration of neuronal dynamics after chronic inactivation of inhibitory neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.17.541057v1?rss=1</link>
<description><![CDATA[
Maintaining motor behaviors throughout life is crucial for an individuals survival and reproductive success. The neuronal mechanisms that preserve behavior are poorly understood. To address this question, we focused on the zebra finch, a bird that produces a highly stereotypical song after learning it as a juvenile. Using cell-specific viral vectors, we chronically silenced inhibitory neurons in the pre-motor song nucleus called the high vocal center (HVC), which caused drastic song degradation. However, after producing severely degraded vocalizations for around 2 months, the song rapidly improved, and animals could sing songs that highly resembled the original. In adult birds, single-cell RNA sequencing of HVC revealed that silencing interneurons elevated markers for microglia and increased expression of the Major Histocompatibility Complex I (MHC I), mirroring changes observed in juveniles during song learning. Interestingly, adults could restore their songs despite lesioning the lateral magnocellular nucleus of the anterior neostriatum (LMAN), a brain nucleus crucial for juvenile song learning. This suggests that while molecular mechanisms may overlap, adults utilize different neuronal mechanisms for song recovery. Chronic and acute electrophysiological recordings within HVC and its downstream target, the robust nucleus of the archistriatum (RA), revealed that neuronal activity in the circuit permanently altered with higher spontaneous firing in RA and lower in HVC compared to control even after the song had fully recovered. Together, our findings show that a complex learned behavior can recover despite extended periods of perturbed behavior and permanently altered neuronal dynamics. These results show that loss of inhibitory tone can be compensated for by recovery mechanisms partly local to the perturbed nucleus and do not require circuits necessary for learning.
]]></description>
<dc:creator>Torok, Z.</dc:creator>
<dc:creator>Luebbert, L.</dc:creator>
<dc:creator>Feldman, J.</dc:creator>
<dc:creator>Duffy, A.</dc:creator>
<dc:creator>Nevue, A.</dc:creator>
<dc:creator>Wongso, S.</dc:creator>
<dc:creator>Mello, C.</dc:creator>
<dc:creator>Fairhall, A.</dc:creator>
<dc:creator>Pachter, L.</dc:creator>
<dc:creator>Gonzalez, W.</dc:creator>
<dc:creator>Lois, C.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.17.541057</dc:identifier>
<dc:title><![CDATA[Recovery of a learned behavior despite partial restoration of neuronal dynamics after chronic inactivation of inhibitory neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.19.541497v1?rss=1">
<title>
<![CDATA[
Engineering Programmable Material-To-Cell Pathways Via Synthetic Notch Receptors To Spatially Control Cellular Phenotypes In Multi-Cellular Constructs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.19.541497v1?rss=1</link>
<description><![CDATA[
Synthetic Notch (synNotch) receptors are modular synthetic components that are genetically engineered into mammalian cells to detect signals presented by neighboring cells and respond by activating prescribed transcriptional programs. To date, synNotch has been used to program therapeutic cells and pattern morphogenesis in multicellular systems. However, cell-presented ligands have limited versatility for applications that require spatial precision, such as tissue engineering. To address this, we developed a suite of materials to activate synNotch receptors and serve as generalizable platforms for generating user-defined material-to-cell signaling pathways. First, we demonstrate that synNotch ligands, such as GFP, can be conjugated to cell- generated ECM proteins via genetic engineering of fibronectin produced by fibroblasts. We then used enzymatic or click chemistry to covalently link synNotch ligands to gelatin polymers to activate synNotch receptors in cells grown on or within a hydrogel. To achieve microscale control over synNotch activation in cell monolayers, we microcontact printed synNotch ligands onto a surface. We also patterned tissues comprising cells with up to three distinct phenotypes by engineering cells with two distinct synthetic pathways and culturing them on surfaces microfluidically patterned with two synNotch ligands.

We showcase this technology by co-transdifferentiating fibroblasts into skeletal muscle or endothelial cell precursors in user-defined spatial patterns towards the engineering of muscle tissue with prescribed vascular networks. Collectively, this suite of approaches extends the synNotch toolkit and provides novel avenues for spatially controlling cellular phenotypes in mammalian multicellular systems, with many broad applications in developmental biology, synthetic morphogenesis, human tissue modeling, and regenerative medicine.
]]></description>
<dc:creator>Garibyan, M.</dc:creator>
<dc:creator>Hoffman, T.</dc:creator>
<dc:creator>Makaske, T.</dc:creator>
<dc:creator>Do, S.</dc:creator>
<dc:creator>March, A. R.</dc:creator>
<dc:creator>Cho, N.</dc:creator>
<dc:creator>Pedroncelli, N.</dc:creator>
<dc:creator>Lima, R. E.</dc:creator>
<dc:creator>Soto, J.</dc:creator>
<dc:creator>Jackson, B. A.</dc:creator>
<dc:creator>Khademhosseini, A.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>McCain, M.</dc:creator>
<dc:creator>Morsut, L.</dc:creator>
<dc:date>2023-05-20</dc:date>
<dc:identifier>doi:10.1101/2023.05.19.541497</dc:identifier>
<dc:title><![CDATA[Engineering Programmable Material-To-Cell Pathways Via Synthetic Notch Receptors To Spatially Control Cellular Phenotypes In Multi-Cellular Constructs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.19.541499v1?rss=1">
<title>
<![CDATA[
The mitochondrial matrix protein cyclophilin D contributes to deficits in parvalbumin interneurons in schizophrenia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.19.541499v1?rss=1</link>
<description><![CDATA[
BACKGROUNDCognitive deficits in schizophrenia are linked to dysfunctions of the dorsolateral prefrontal cortex (DLPFC), including alterations in parvalbumin (PV)-expressing interneurons (PVIs). Redox dysregulation and oxidative stress may represent convergence points in the pathology of schizophrenia, causing dysfunction of GABAergic interneurons and loss of PV. Here, we show that the mitochondrial matrix protein cyclophilin-D (CypD), a critical initiator of the mitochondrial permeability transition pore (mPTP) and modulator of the intracellular redox state, is altered in PVIs in schizophrenia.

METHODSWestern blotting was used to measure CypD protein levels in postmortem DLPFC specimens of schizophrenic (n=27) and matched comparison subjects with no known history of psychiatric or neurological disorders (n=26). In a subset of this cohort, multilabel immunofluorescent confocal microscopy with unbiased stereological sampling methods were used to quantify 1) numbers of PVI across the cortical mantle (20 control, 15 schizophrenia) and 2) PV and CypD protein levels from PVIs in the cortical layers 2-4 (23 control, 19 schizophrenia).

RESULTSIn schizophrenic subjects, the overall number of PVIs in the DLPFC was not significantly altered, but in individual PVIs of layers 2-4 PV protein levels decreased along a superficial-to-deep gradient when compared to unaffected comparison subjects. These laminar-specific PVI alterations were reciprocally linked to significant CypD elevations both in PVIs and within total DLPFC gray matter.

CONCLUSIONSOur findings support previously reported PVI anomalies in schizophrenia and suggest that CypD-mediated mPTP formation could be a potential contributor to PVI dysfunction in schizophrenia.
]]></description>
<dc:creator>O'Brien, J. T.</dc:creator>
<dc:creator>Jalilvand, S. P.</dc:creator>
<dc:creator>Suji, N. A.</dc:creator>
<dc:creator>Phensy, A.</dc:creator>
<dc:creator>Jupelly, R. K.</dc:creator>
<dc:creator>Mwirigi, J. M.</dc:creator>
<dc:creator>Elahi, H.</dc:creator>
<dc:creator>Price, T. J.</dc:creator>
<dc:creator>Kroener, S.</dc:creator>
<dc:date>2023-05-20</dc:date>
<dc:identifier>doi:10.1101/2023.05.19.541499</dc:identifier>
<dc:title><![CDATA[The mitochondrial matrix protein cyclophilin D contributes to deficits in parvalbumin interneurons in schizophrenia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.20.541529v1?rss=1">
<title>
<![CDATA[
Chemoenzymatic syntheses of fluorine-18-labeled disaccharides from FDG yield potent sensors of living bacteria in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.20.541529v1?rss=1</link>
<description><![CDATA[
Chemoenzymatic techniques have been applied extensively to pharmaceutical development, most effectively when routine synthetic methods fail. The regioselective and stereoselective construction of structurally complex glycans is an elegant application of this approach, that is seldom applied to positron emission tomography (PET) tracers. We sought a method to dimerize 2-deoxy-[18F]-fluoro-D-glucose ([18F]FDG), the most common tracer used in clinical imaging, to form [18F]-labeled disaccharides for detecting microorganisms in vivo based on their bacteria-specific glycan incorporation. When [18F]FDG was reacted with {beta}-D-glucose-1-phosphate in the presence of maltose phosphorylase, both the -1,4 and -1,3-linked products 2-deoxy-[18F]-fluoro-maltose ([18F]FDM) and 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK) were obtained. This method was further extended with the use of trehalose (,-1,1), laminaribiose ({beta}-1,3), and cellobiose ({beta}-1,4) phosphorylases to synthesize 2-deoxy-2-[18F]fluoro-trehalose ([18F]FDT), 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), and 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC). We subsequently tested [18F]FDM and [18F]FSK in vitro, showing accumulation by several clinically relevant pathogens including Staphylococcus aureus and Acinetobacter baumannii, and demonstrated their specific uptake in vivo. The lead sakebiose-derived tracer [18F]FSK was stable in human serum and showed high uptake in preclinical models of myositis and vertebral discitis-osteomyelitis. Both the synthetic ease, and high sensitivity of [18F]FSK to S. aureus including methicillin-resistant (MRSA) strains strongly justify clinical translation of this tracer to infected patients. Furthermore, this work suggests that chemoenzymatic radiosyntheses of complex [18F]FDG-derived oligomers will afford a wide array of PET radiotracers for infectious and oncologic applications.
]]></description>
<dc:creator>Sorlin, A.</dc:creator>
<dc:creator>Lopez Alvarez, M.</dc:creator>
<dc:creator>Rabbitt, S.</dc:creator>
<dc:creator>Alanizi, A.</dc:creator>
<dc:creator>Shuere, B.</dc:creator>
<dc:creator>Bobba, K. N.</dc:creator>
<dc:creator>Blecha, J.</dc:creator>
<dc:creator>Sakhamuri, S.</dc:creator>
<dc:creator>Evans, M.</dc:creator>
<dc:creator>Bayles, K. J.</dc:creator>
<dc:creator>Flavell, R.</dc:creator>
<dc:creator>Rosenberg, O.</dc:creator>
<dc:creator>Sriram, R.</dc:creator>
<dc:creator>Desmet, T.</dc:creator>
<dc:creator>Nidetzky, B.</dc:creator>
<dc:creator>Engel, J.</dc:creator>
<dc:creator>Ohliger, M.</dc:creator>
<dc:creator>Fraser, J.</dc:creator>
<dc:creator>Wilson, D. M.</dc:creator>
<dc:date>2023-05-20</dc:date>
<dc:identifier>doi:10.1101/2023.05.20.541529</dc:identifier>
<dc:title><![CDATA[Chemoenzymatic syntheses of fluorine-18-labeled disaccharides from FDG yield potent sensors of living bacteria in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.18.541379v1?rss=1">
<title>
<![CDATA[
Neurophysiological trajectories in Alzheimer's disease progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.18.541379v1?rss=1</link>
<description><![CDATA[
Alzheimers disease (AD) is characterized by the accumulation of amyloid-{beta} and misfolded tau proteins causing synaptic dysfunction, and progressive neurodegeneration and cognitive decline. Altered neural oscillations have been consistently demonstrated in AD. However, the trajectories of abnormal neural oscillations in AD progression and their relationship to neurodegeneration and cognitive decline are unknown. Here, we deployed robust event-based sequencing models (EBMs) to investigate the trajectories of long-range and local neural synchrony across AD stages, estimated from resting-state magnetoencephalography. The increases in neural synchrony in the delta-theta band and the decreases in the alpha and beta bands showed progressive changes throughout the stages of the EBM. Decreases in alpha and beta band synchrony preceded both neurodegeneration and cognitive decline, indicating that frequency-specific neuronal synchrony abnormalities are early manifestations of AD pathophysiology. The long-range synchrony effects were greater than the local synchrony, indicating a greater sensitivity of connectivity metrics involving multiple regions of the brain. These results demonstrate the evolution of functional neuronal deficits along the sequence of AD progression.
]]></description>
<dc:creator>Kudo, K.</dc:creator>
<dc:creator>Ranasinghe, K. G.</dc:creator>
<dc:creator>Morise, H.</dc:creator>
<dc:creator>Syed, F.</dc:creator>
<dc:creator>Sekihara, K.</dc:creator>
<dc:creator>Rankin, K. P.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Rabinovici, G.</dc:creator>
<dc:creator>Vossel, K.</dc:creator>
<dc:creator>Kirsch, H. E.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.18.541379</dc:identifier>
<dc:title><![CDATA[Neurophysiological trajectories in Alzheimer's disease progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.19.541376v1?rss=1">
<title>
<![CDATA[
A Scalable High Throughput Fully Automated Pipeline for the Quantification of Amyloid Pathology in Alzheimer's Disease Using Deep Learning Algorithms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.19.541376v1?rss=1</link>
<description><![CDATA[
The most common approach to characterize neuropathology in Alzheimers disease (AD) involves a manual survey and inspection by an expert neuropathologist of postmortem tissue that has been immunolabeled to visualize the presence of amyloid {beta} in plaques and around blood vessels and neurofibrillary tangles of the tau protein. In the case of amyloid {beta} pathology, a semiquantitative score is given that is based on areas of densest pathology. The approach has been well-validated but the process is laborious and time consuming, and inherently susceptible to intra- and inter-observer variability. Moreover, the tremendous growth in genetic, transcriptomic and proteomic data from AD patients has created new opportunities to link clinical features of AD to molecular pathogenesis through pathology, but the lack of high throughput quantitative and comprehensive approaches to assess neuropathology limits the associations that can be discovered. To address these limitations, we designed a computational pipeline to analyze postmortem tissue from AD patients in a fully automated, unbiased and high throughput manner. We used deep learning to train algorithms with a Mask Regional-Convolutional Neural Network to detect and classify different types of amyloid pathology with human level accuracy. After training on pathology slides from a Mt Sinai cohort, our algorithms identified amyloid pathology in samples made at an independent brain bank and from an unrelated cohort of patients, indicating that the algorithms were detecting reproducible and generalizable pathology features. We designed the pipeline to retain the position of the pathology it detects, making it possible to reconstruct a map of pathology across the entire whole slide image, facilitating neuropathological analyses at multiple scales. Quantitative measurements of amyloid pathology correlated positively and significantly with the severity of AD as measured by standard approaches. We conclude that we have developed a computational pipeline to analyze digitized images of neuropathology in high throughput and algorithms to detect types of amyloid pathology with human level accuracy that should enable neuropathological analysis of large tissue collections and integration of those results with orthogonal clinical and multiomic measurements.
]]></description>
<dc:creator>Gopal Ramaswamy, V.</dc:creator>
<dc:creator>Ahirwar, M.</dc:creator>
<dc:creator>Ryan, G.</dc:creator>
<dc:creator>Signaevski, M.</dc:creator>
<dc:creator>Haroutunian, V.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.19.541376</dc:identifier>
<dc:title><![CDATA[A Scalable High Throughput Fully Automated Pipeline for the Quantification of Amyloid Pathology in Alzheimer's Disease Using Deep Learning Algorithms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.22.541795v1?rss=1">
<title>
<![CDATA[
Identification of an anti-CRISPR protein that inhibits the CRISPR-Cas type I-B system in Clostridioides difficile 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.22.541795v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas systems provide their prokaryotic hosts with adaptive immunity against mobile genetic elements. Many bacteriophages encode anti-CRISPR (Acr) proteins that inhibit host defense. The identification of Acr proteins is challenging due to their small size and high sequence diversity, and only a limited number has been characterized to date. In this study, we report the discovery of a novel Acr protein, AcrIB2, encoded by the {varphi}CD38-2 Clostridioides difficile phage that efficiently inhibits interference by the type I-B CRISPR-Cas system of the host and likely acts as a DNA mimic. Most C. difficile strains contain two cas operons, one encoding a full set of interference and adaptation proteins and another encoding interference proteins only. Unexpectedly, we show that only the partial operon is required for interference and is subject to inhibition by AcrIB2.
]]></description>
<dc:creator>Muzyukina, P.</dc:creator>
<dc:creator>Shkaruta, A.</dc:creator>
<dc:creator>Guzman, N. M.</dc:creator>
<dc:creator>Andreani, J.</dc:creator>
<dc:creator>Borges, A. L.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Maikova, A.</dc:creator>
<dc:creator>Semenova, E.</dc:creator>
<dc:creator>Severinov, K.</dc:creator>
<dc:creator>SOUTOURINA, O.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.22.541795</dc:identifier>
<dc:title><![CDATA[Identification of an anti-CRISPR protein that inhibits the CRISPR-Cas type I-B system in Clostridioides difficile]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.22.541477v1?rss=1">
<title>
<![CDATA[
Long-term optical imaging of the spinal cord in awake, behaving animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.22.541477v1?rss=1</link>
<description><![CDATA[
Advances in optical imaging approaches and fluorescent biosensors have enabled an understanding of the spatiotemporal and long-term neural dynamics in the brain of awake animals. However, methodological difficulties and the persistence of post-laminectomy fibrosis have greatly limited similar advances in the spinal cord. To overcome these technical obstacles, we combined in vivo application of fluoropolymer membranes that inhibit fibrosis; a redesigned, cost-effective implantable spinal imaging chamber; and improved motion correction methods that together permit imaging of the spinal cord in awake, behaving mice, for months to over a year. We also demonstrate a robust ability to monitor axons, identify a spinal cord somatotopic map, conduct Ca2+ imaging of neural dynamics in behaving animals responding to pain-provoking stimuli, and observe persistent microglial changes after nerve injury. The ability to couple neural activity and behavior at the spinal cord level will drive insights not previously possible at a key location for somatosensory transmission to the brain.
]]></description>
<dc:creator>Ahanonu, B.</dc:creator>
<dc:creator>Crowther, A.</dc:creator>
<dc:creator>Kania, A.</dc:creator>
<dc:creator>Casillas, M. R.</dc:creator>
<dc:creator>Basbaum, A.</dc:creator>
<dc:date>2023-05-24</dc:date>
<dc:identifier>doi:10.1101/2023.05.22.541477</dc:identifier>
<dc:title><![CDATA[Long-term optical imaging of the spinal cord in awake, behaving animals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.23.540554v1?rss=1">
<title>
<![CDATA[
GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.23.540554v1?rss=1</link>
<description><![CDATA[
The orphan G protein-coupled receptor (GPCR) GPR161 is enriched in primary cilia, where it plays a central role in suppressing Hedgehog signaling1. GPR161 mutations lead to developmental defects and cancers2,3,4. The fundamental basis of how GPR161 is activated, including potential endogenous activators and pathway-relevant signal transducers, remains unclear. To elucidate GPR161 function, we determined a cryogenic-electron microscopy structure of active GPR161 bound to the heterotrimeric G protein complex Gs. This structure revealed an extracellular loop 2 that occupies the canonical GPCR orthosteric ligand pocket. Furthermore, we identify a sterol that binds to a conserved extrahelical site adjacent to transmembrane helices 6 and 7 and stabilizes a GPR161 conformation required for Gs coupling. Mutations that prevent sterol binding to GPR161 suppress cAMP pathway activation. Surprisingly, these mutants retain the ability to suppress GLI2 transcription factor accumulation in cilia, a key function of ciliary GPR161 in Hedgehog pathway suppression. By contrast, a protein kinase A-binding site in the GPR161 C-terminus is critical in suppressing GLI2 ciliary accumulation. Our work highlights how unique structural features of GPR161 interface with the Hedgehog pathway and sets a foundation to understand the broader role of GPR161 function in other signaling pathways.
]]></description>
<dc:creator>Hoppe, N.</dc:creator>
<dc:creator>Harrison, S.</dc:creator>
<dc:creator>Hwang, S.-H.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Karelina, M.</dc:creator>
<dc:creator>Deshpande, I.</dc:creator>
<dc:creator>Suomivuori, C.-M.</dc:creator>
<dc:creator>Palicharla, V. R.</dc:creator>
<dc:creator>Berry, S. P.</dc:creator>
<dc:creator>Tschaikner, P.</dc:creator>
<dc:creator>Regele, D.</dc:creator>
<dc:creator>Covey, D. F.</dc:creator>
<dc:creator>Stefan, E.</dc:creator>
<dc:creator>Marks, D. S.</dc:creator>
<dc:creator>Reiter, J.</dc:creator>
<dc:creator>Dror, R. O.</dc:creator>
<dc:creator>Evers, A. S.</dc:creator>
<dc:creator>Mukhopadhyay, S.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:date>2023-05-24</dc:date>
<dc:identifier>doi:10.1101/2023.05.23.540554</dc:identifier>
<dc:title><![CDATA[GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.24.542054v1?rss=1">
<title>
<![CDATA[
Comparison of histological delineations of medial temporal lobe cortices by four independent neuroanatomy laboratories 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.24.542054v1?rss=1</link>
<description><![CDATA[
The medial temporal lobe (MTL) cortex, located adjacent to the hippocampus, is crucial for memory and prone to the accumulation of certain neuropathologies such as Alzheimers disease neurofibrillary tau tangles. The MTL cortex is composed of several subregions which differ in their functional and cytoarchitectonic features. As neuroanatomical schools rely on different cytoarchitectonic definitions of these subregions, it is unclear to what extent their delineations of MTL cortex subregions overlap. Here, we provide an overview of cytoarchitectonic definitions of the cortices that make up the parahippocampal gyrus (entorhinal and parahippocampal cortices) and the adjacent Brodmann areas (BA) 35 and 36, as provided by four neuroanatomists from different laboratories, aiming to identify the rationale for overlapping and diverging delineations.

Nissl-stained series were acquired from the temporal lobes of three human specimens (two right and one left hemisphere). Slices (50 {micro}m thick) were prepared perpendicular to the long axis of the hippocampus spanning the entire longitudinal extent of the MTL cortex. Four neuroanatomists annotated MTL cortex subregions on digitized (20X resolution) slices with 5 mm spacing. Parcellations, terminology, and border placement were compared among neuroanatomists. Cytoarchitectonic features of each subregion are described in detail.

Qualitative analysis of the annotations showed higher agreement in the definitions of the entorhinal cortex and BA35, while definitions of BA36 and the parahippocampal cortex exhibited less overlap among neuroanatomists. The degree of overlap of cytoarchitectonic definitions was partially reflected in the neuroanatomists agreement on the respective delineations. Lower agreement in annotations was observed in transitional zones between structures where seminal cytoarchitectonic features are expressed more gradually.

The results highlight that definitions and parcellations of the MTL cortex differ among neuroanatomical schools and thereby increase understanding of why these differences may arise. This work sets a crucial foundation to further advance anatomically-informed human neuroimaging research on the MTL cortex.
]]></description>
<dc:creator>Wuestefeld, A.</dc:creator>
<dc:creator>Baumeister, H.</dc:creator>
<dc:creator>Adams, J. N.</dc:creator>
<dc:creator>de Flores, R.</dc:creator>
<dc:creator>Hodgetts, C.</dc:creator>
<dc:creator>Mazloum-Farzaghi, N.</dc:creator>
<dc:creator>Olsen, R. K.</dc:creator>
<dc:creator>Puliyadi, V.</dc:creator>
<dc:creator>Tran, T. T.</dc:creator>
<dc:creator>Bakker, A.</dc:creator>
<dc:creator>Canada, K. L.</dc:creator>
<dc:creator>Dalton, M. A.</dc:creator>
<dc:creator>Daugherty, A. M.</dc:creator>
<dc:creator>La Joie, R.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Bedard, M.</dc:creator>
<dc:creator>Buendia, E.</dc:creator>
<dc:creator>Denning, A.</dc:creator>
<dc:creator>Irwin, D. J.</dc:creator>
<dc:creator>Ittyerah, R.</dc:creator>
<dc:creator>Lee, E. B.</dc:creator>
<dc:creator>Lim, S.</dc:creator>
<dc:creator>Iniguez de Onzono Martin, M. M.</dc:creator>
<dc:creator>Munoz Lopez, M.</dc:creator>
<dc:creator>Schuck, T.</dc:creator>
<dc:creator>Trotman, W.</dc:creator>
<dc:creator>Vela, A.</dc:creator>
<dc:creator>Yushkevich, P.</dc:creator>
<dc:creator>Amunts, K.</dc:creator>
<dc:creator>Augustinack, J. C.</dc:creator>
<dc:creator>Ding, S.-L.</dc:creator>
<dc:creator>Insausti, R.</dc:creator>
<dc:creator>Kedo, O.</dc:creator>
<dc:creator>Berron, D.</dc:creator>
<dc:creator>Wisse, L. E. M.</dc:creator>
<dc:date>2023-05-24</dc:date>
<dc:identifier>doi:10.1101/2023.05.24.542054</dc:identifier>
<dc:title><![CDATA[Comparison of histological delineations of medial temporal lobe cortices by four independent neuroanatomy laboratories]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.25.542288v1?rss=1">
<title>
<![CDATA[
Type-2 diabetes alters hippocampal neural oscillations and disrupts synchrony between hippocampus and cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.25.542288v1?rss=1</link>
<description><![CDATA[
Type 2 diabetes mellitus (T2DM) increases the risk of neurological diseases, yet how brain oscillations change as age and T2DM interact is not well characterized. To delineate the age and diabetic effect on neurophysiology, we recorded local field potentials with multichannel electrodes spanning the somatosensory cortex and hippocampus (HPC) under urethane anesthesia in diabetic and normoglycemic control mice, at 200 and 400 days of age. We analyzed the signal power of brain oscillations, brain state, sharp wave associate ripples (SPW-Rs), and functional connectivity between the cortex and HPC. We found that while both age and T2DM were correlated with a breakdown in long-range functional connectivity and reduced neurogenesis in the dentate gyrus and subventricular zone, T2DM further slowed brain oscillations and reduced theta-gamma coupling. Age and T2DM also prolonged the duration of SPW-Rs and increased gamma power during SPW-R phase. Our results have identified potential electrophysiological substrates of hippocampal changes associated with T2DM and age. The perturbed brain oscillation features and diminished neurogenesis may underlie T2DM-accelerated cognitive impairment.
]]></description>
<dc:creator>Rabiller, G.</dc:creator>
<dc:creator>Ip, Z.</dc:creator>
<dc:creator>Zarrabian, S.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Sato, Y.</dc:creator>
<dc:creator>Yazdan-Shahmorad, A.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:date>2023-05-26</dc:date>
<dc:identifier>doi:10.1101/2023.05.25.542288</dc:identifier>
<dc:title><![CDATA[Type-2 diabetes alters hippocampal neural oscillations and disrupts synchrony between hippocampus and cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.25.542333v1?rss=1">
<title>
<![CDATA[
Contrasting synaptic roles of MDGA1 and MDGA2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.25.542333v1?rss=1</link>
<description><![CDATA[
Neurodevelopmental disorders are frequently linked to mutations in synaptic organizing molecules. MAM domain containing glycosylphosphatidylinositol anchor 1 and 2 (MDGA1 and MDGA2) are a family of synaptic organizers suggested to play an unusual role as synaptic repressors, but studies offer conflicting evidence for their localization. Using epitope-tagged MDGA1 and MDGA2 knock-in mice, we found that native MDGAs are expressed throughout the brain, peaking early in postnatal development. Surprisingly, endogenous MDGA1 was enriched at excitatory, but not inhibitory, synapses. Both shRNA knockdown and CRISPR/Cas9 knockout of MDGA1 resulted in cell-autonomous, specific impairment of AMPA receptor- mediated synaptic transmission, without affecting GABAergic transmission. Conversely, MDGA2 knockdown/knockout selectively depressed NMDA receptor-mediated transmission but enhanced inhibitory transmission. Our results establish that MDGA2 acts as a synaptic repressor, but only at inhibitory synapses, whereas both MDGAs are required for excitatory transmission. This nonoverlapping division of labor between two highly conserved synaptic proteins is unprecedented.

TeaserMDGAs 1 and 2 independently localize to and modulate excitatory and inhibitory hippocampal synapses by different mechanisms.
]]></description>
<dc:creator>Bemben, M. A.</dc:creator>
<dc:creator>Sandoval, M. A.</dc:creator>
<dc:creator>Le, A. A.</dc:creator>
<dc:creator>Won, S.</dc:creator>
<dc:creator>Chau, V. N.</dc:creator>
<dc:creator>Lauterborn, J. C.</dc:creator>
<dc:creator>Incontro, S.</dc:creator>
<dc:creator>Li, K. H.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Roche, K. W.</dc:creator>
<dc:creator>Gall, C. M.</dc:creator>
<dc:creator>Nicoll, R. A.</dc:creator>
<dc:creator>Diaz-Alonso, J.</dc:creator>
<dc:date>2023-05-26</dc:date>
<dc:identifier>doi:10.1101/2023.05.25.542333</dc:identifier>
<dc:title><![CDATA[Contrasting synaptic roles of MDGA1 and MDGA2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.26.542293v1?rss=1">
<title>
<![CDATA[
Association of an estrogen-sensitive Pax1-Col11a1-Mmp3 signaling axis with adolescent idiopathic scoliosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.26.542293v1?rss=1</link>
<description><![CDATA[
Adolescent idiopathic scoliosis (AIS) is a common and progressive spinal deformity in children that exhibits striking sexual dimorphism, with girls at more than five-fold greater risk of severe disease compared to boys. Despite its medical impact, the molecular mechanisms that drive AIS are largely unknown. We previously defined a female-specific AIS genetic risk locus in an enhancer near the PAX1 gene. Here we sought to define the roles of PAX1 and newly-identified AIS-associated genes in the developmental mechanism of AIS. In a genetic study of 10,519 individuals with AIS and 93,238 unaffected controls, significant association was identified with a variant in COL11A1 encoding collagen (1) XI (rs3753841; NM_080629.2_c.4004C>T; p.(Pro1335Leu); P=7.07e-11, OR=1.118). Using CRISPR mutagenesis we generated Pax1 knockout mice (Pax1-/-). In postnatal spines we found that PAX1 and collagen (1) XI protein both localize within the intervertebral disc (IVD)-vertebral junction region encompassing the growth plate, with less collagen (1) XI detected in Pax1-/- spines compared to wildtype. By genetic targeting we found that wildtype Col11a1 expression in costal chondrocytes suppresses expression of Pax1 and of Mmp3, encoding the matrix metalloproteinase 3 enzyme implicated in matrix remodeling. However, this suppression was abrogated in the presence of the AIS-associated COL11A1P1335L mutant. Further, we found that either knockdown of the estrogen receptor gene Esr2, or tamoxifen treatment, significantly altered Col11a1 and Mmp3 expression in chondrocytes. We propose a new molecular model of AIS pathogenesis wherein genetic variation and estrogen signaling increase disease susceptibility by altering a Pax1-Col11a1-Mmp3 signaling axis in spinal chondrocytes.
]]></description>
<dc:creator>Wise, C.</dc:creator>
<dc:creator>Yu, H.</dc:creator>
<dc:creator>Khanshour, A. M.</dc:creator>
<dc:creator>Ushiki, A.</dc:creator>
<dc:creator>Otomo, N.</dc:creator>
<dc:creator>Koike, Y.</dc:creator>
<dc:creator>Einarsdottir, E.</dc:creator>
<dc:creator>Fan, Y.</dc:creator>
<dc:creator>Antunes, L.</dc:creator>
<dc:creator>Kidane, Y. H.</dc:creator>
<dc:creator>Sheng, R.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Pei, J.</dc:creator>
<dc:creator>Grishin, N. V.</dc:creator>
<dc:creator>Evers, B. M. V.</dc:creator>
<dc:creator>Cheung, J. P. Y.</dc:creator>
<dc:creator>Herring, J. A.</dc:creator>
<dc:creator>Terao, C.</dc:creator>
<dc:creator>Song, Y.-Q.</dc:creator>
<dc:creator>Gurnett, C.</dc:creator>
<dc:creator>Gerdhem, P.</dc:creator>
<dc:creator>Ikegawa, S.</dc:creator>
<dc:creator>Rios, J. J.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:date>2023-05-27</dc:date>
<dc:identifier>doi:10.1101/2023.05.26.542293</dc:identifier>
<dc:title><![CDATA[Association of an estrogen-sensitive Pax1-Col11a1-Mmp3 signaling axis with adolescent idiopathic scoliosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.27.542570v1?rss=1">
<title>
<![CDATA[
Expression and purification of the mitochondrial transmembrane protein FAM210A in Escherichia coli 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.27.542570v1?rss=1</link>
<description><![CDATA[
The protein Family with sequence similarity 210 member A (FAM210A) is a mitochondrial inner membrane protein that regulates the protein synthesis of mitochondrial DNA encoded genes. However, how it functions in this process is not well understood. Developing and optimizing a protein purification strategy will facilitate biochemical and structural studies of FAM210A. Here, we developed a method to purify human FAM210A with deleted mitochondrial targeting signal sequence using the MBP-His10 fusion in Escherichia coli. The recombinant FAM210A protein was inserted into the E. coli cell membrane and purified from isolated bacterial cell membranes, followed by a two-step process using Ni-NTA resin-based immobilized-metal affinity chromatography (IMAC) and ion exchange purification. A pulldown assay validated the functionality of purified FAM210A protein interacting with human mitochondrial elongation factor EF-Tu in HEK293T cell lysates. Taken together, this study developed a method for purification of the mitochondrial transmembrane protein FAM210A partially complexed with E.coli derived EF-Tu and provides an opportunity for future potential biochemical and structural studies of recombinant FAM210A protein.
]]></description>
<dc:creator>Hollinger, J.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Awayda, K. M.</dc:creator>
<dc:creator>O'Connell, M. R.</dc:creator>
<dc:creator>Yao, P.</dc:creator>
<dc:date>2023-05-27</dc:date>
<dc:identifier>doi:10.1101/2023.05.27.542570</dc:identifier>
<dc:title><![CDATA[Expression and purification of the mitochondrial transmembrane protein FAM210A in Escherichia coli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.26.542486v1?rss=1">
<title>
<![CDATA[
Coupling and uncoupling of midline morphogenesis and cell flow in amniote gastrulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.26.542486v1?rss=1</link>
<description><![CDATA[
Large-scale cell flow characterizes gastrulation in animal development. In amniote gastrulation, particularly in avian gastrula, a bilateral vortex-like counter-rotating cell flow, called  polonaise movements, appears along the midline. Here, through experimental manipulations, we addressed relationships between the polonaise movements and morphogenesis of the primitive streak, the earliest midline structure in amniotes. Suppression of the Wnt/planar cell polarity (PCP) signaling pathway maintains the polonaise movements along a deformed primitive streak. Mitotic arrest leads to diminished extension and development of the primitive streak and maintains the early phase of the polonaise movements. Ectopically induced Vg1, an axis-inducing morphogen, generates the polonaise movements, aligned to the induced midline, but disturbs the stereotypical cell flow pattern at the authentic midline. Despite the altered cell flow, induction and extension of the primitive streak are preserved along both authentic and induced midlines. Finally, we show that ectopic axis-inducing morphogen, Vg1, is capable of initiating the polonaise movements without concomitant PS extension under mitotic arrest conditions. These results are consistent with a model wherein primitive streak morphogenesis is required for the maintenance of the polonaise movements, but the polonaise movements are not necessarily responsible for primitive streak morphogenesis. Our data describe a previously undefined relationship between the large-scale cell flow and midline morphogenesis in gastrulation.
]]></description>
<dc:creator>Asai, R.</dc:creator>
<dc:creator>Prakash, V. N.</dc:creator>
<dc:creator>Sinha, S.</dc:creator>
<dc:creator>Prakash, M.</dc:creator>
<dc:creator>Mikawa, T.</dc:creator>
<dc:date>2023-05-28</dc:date>
<dc:identifier>doi:10.1101/2023.05.26.542486</dc:identifier>
<dc:title><![CDATA[Coupling and uncoupling of midline morphogenesis and cell flow in amniote gastrulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.27.542563v1?rss=1">
<title>
<![CDATA[
Sequential Switching Through IgG1 is Redundant for Allergic Reactivity and Memory to Allergens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.27.542563v1?rss=1</link>
<description><![CDATA[
Allergic reactions to foods are driven by allergen-binding immunoglobulin (Ig)E antibodies. IgE- expressing cells can be generated through a sequential class switching pathway where activated B cells first switch to an intermediary isotype, most frequently IgG1, and then to IgE. It has been proposed that sequential class switch recombination is important in generating high affinity IgE, augmenting anaphylactic reactions, and in holding the memory of IgE responses. Here, we observed surprising redundancy of sequential switching through IgG1 for the functional affinity of the IgE repertoire against multiple food allergens as well as for the ability of IgE to elicit anaphylaxis. We further found that sequential switching via IgG1 was irrelevant for allergic memory. These results indicate that allergen-specific IgG1 B cells are redundant in sensitization, anaphylaxis, and food allergy persistence, thereby implicating other switching pathways as important considerations in the development of therapeutics for allergic diseases.
]]></description>
<dc:creator>Koenig, J. F.</dc:creator>
<dc:creator>Wade-Vallance, A. K.</dc:creator>
<dc:creator>Jimenez-Saiz, R.</dc:creator>
<dc:creator>Bruton, K.</dc:creator>
<dc:creator>Gadkar, S.</dc:creator>
<dc:creator>Grydziuszko, E.</dc:creator>
<dc:creator>Walker, T.</dc:creator>
<dc:creator>Gordon, M. E.</dc:creator>
<dc:creator>Waserman, S.</dc:creator>
<dc:creator>Jordana, M.</dc:creator>
<dc:date>2023-05-28</dc:date>
<dc:identifier>doi:10.1101/2023.05.27.542563</dc:identifier>
<dc:title><![CDATA[Sequential Switching Through IgG1 is Redundant for Allergic Reactivity and Memory to Allergens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.29.542707v1?rss=1">
<title>
<![CDATA[
Exomap1 mouse: a transgenic model for in vivo studies of exosome biology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.29.542707v1?rss=1</link>
<description><![CDATA[
Exosomes are small extracellular vesicles (sEVs) of [~]30-150 nm in diameter that have the same topology as the cell, are enriched in selected exosome cargo proteins, and play important roles in health and disease. To address large unanswered questions regarding exosome biology in vivo, we created the exomap1 transgenic mouse model. In response to Cre recombinase, exomap1 mice express HsCD81mNG, a fusion protein between human CD81, the most highly enriched exosome protein yet described, and the bright green fluorescent protein mNeonGreen. As expected, cell type-specific expression of Cre induced the cell type-specific expression of HsCD81mNG in diverse cell types, correctly localized HsCD81mNG to the plasma membrane, and selectively loaded HsCD81mNG into secreted vesicles that have the size ([~]80 nm), topology (outside out), and content (presence of mouse exosome markers) of exosomes. Furthermore, mouse cells expressing HsCD81mNG released HsCD81mNG-marked exosomes into blood and other biofluids. Using high-resolution, single-exosome analysis by quantitative single molecule localization microscopy, we show here that that hepatocytes contribute [~]15% of the blood exosome population whereas neurons contribute <1% of blood exosomes. These estimates of cell type-specific contributions to blood EV population are consistent with the porosity of liver sinusoidal endothelial cells to particles of [~]50-300 nm in diameter, as well as with the impermeability of blood-brain and blood-neuron barriers to particles >5 nm in size. Taken together, these results establish the exomap1 mouse as a useful tool for in vivo studies of exosome biology, and for mapping cell type-specific contributions to biofluid exosome populations. In addition, our data confirm that CD81 is a highly-specific marker for exosomes and is not enriched in the larger microvesicle class of EVs.
]]></description>
<dc:creator>Fordjour, F. K.</dc:creator>
<dc:creator>Abuelreich, S.</dc:creator>
<dc:creator>Hong, X.</dc:creator>
<dc:creator>Chatterjee, E.</dc:creator>
<dc:creator>Lallai, V.</dc:creator>
<dc:creator>Ng, M.</dc:creator>
<dc:creator>Saftics, A.</dc:creator>
<dc:creator>Deng, F.</dc:creator>
<dc:creator>Carnel-Amar, N.</dc:creator>
<dc:creator>Wakimoto, H.</dc:creator>
<dc:creator>Shimizu, K.</dc:creator>
<dc:creator>Anh Phu, T.</dc:creator>
<dc:creator>Vu, N. K.</dc:creator>
<dc:creator>Geiger, P. C.</dc:creator>
<dc:creator>Raffai, R. L.</dc:creator>
<dc:creator>Fowler, C. D.</dc:creator>
<dc:creator>Shimizu, K.</dc:creator>
<dc:creator>Das, S.</dc:creator>
<dc:creator>Christenson, L. K.</dc:creator>
<dc:creator>Jovanovic-Talisman, T.</dc:creator>
<dc:creator>Gould, S.</dc:creator>
<dc:date>2023-05-29</dc:date>
<dc:identifier>doi:10.1101/2023.05.29.542707</dc:identifier>
<dc:title><![CDATA[Exomap1 mouse: a transgenic model for in vivo studies of exosome biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.30.542893v1?rss=1">
<title>
<![CDATA[
The oncogene cyclin D1 promotes bipolar spindle integrity under compressive force 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.30.542893v1?rss=1</link>
<description><![CDATA[
The mitotic spindle is the bipolar, microtubule-based structure that segregates chromosomes at each cell division. Aberrant spindles are frequently observed in cancer cells, but how oncogenic transformation affects spindle mechanics and function, particularly in the mechanical context of solid tumors, remains poorly understood. Here, we constitutively overexpress the oncogene cyclin D1 in human MCF10A cells to probe its effects on spindle architecture and response to compressive force. We find that cyclin D1 overexpression increases the incidence of spindles with extra poles, centrioles, and chromosomes. However, it also protects spindle poles from fracturing under compressive force, a deleterious outcome linked to multipolar cell divisions. Our findings suggest that cyclin D1 overexpression may adapt cells to increased compressive stress, contributing to its prevalence in cancers such as breast cancer by allowing continued proliferation in mechanically challenging environments.
]]></description>
<dc:creator>Sutanto, R.</dc:creator>
<dc:creator>Neahring, L.</dc:creator>
<dc:creator>Marques, A. S.</dc:creator>
<dc:creator>Kilinc, S.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2023-05-31</dc:date>
<dc:identifier>doi:10.1101/2023.05.30.542893</dc:identifier>
<dc:title><![CDATA[The oncogene cyclin D1 promotes bipolar spindle integrity under compressive force]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.29.542756v1?rss=1">
<title>
<![CDATA[
Reference-free multiplexed single-cell sequencing identifies genetic modifiers of the human immune response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.29.542756v1?rss=1</link>
<description><![CDATA[
Multiplexed single-cell sequencing (mux-seq) using single-nucleotide polymorphisms (SNPs) has emerged as an efficient approach to perform expression quantitative trait loci (eQTL) studies that map interactions between genetic variants and cell types, cell states, or experimental perturbations. Here we introduce the clue framework, a novel approach to encode mux-seq experiments that eliminates the need for reference genotypes and experimental barcoding. The clue framework is made possible by the development of freemuxlet, an algorithm that clusters cells based on SNPs called from single-cell RNA-seq or ATAC-seq data. To demonstrate the feasibility of clue, we profiled the surface protein and RNA abundances of peripheral blood mononuclear cells from 64 individuals, stimulated with 5 distinct extracellular stimuli -- all within a single day. Our analysis of the demultiplexed data identified rare immune cell types and cell type-specific responses to interferon and toll-like receptor stimulation. Furthermore, by integrating genotyping data, we mapped response eQTLs specific to certain cell types. These findings showcase the potential and scalability of the clue framework for reference-free multiplexed single-cell sequencing studies.
]]></description>
<dc:creator>Hartoularos, G. C.</dc:creator>
<dc:creator>Si, Y.</dc:creator>
<dc:creator>Zhang, F. C.</dc:creator>
<dc:creator>Kathail, P.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Ogorodnikov, A.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Kang, H. M.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.05.29.542756</dc:identifier>
<dc:title><![CDATA[Reference-free multiplexed single-cell sequencing identifies genetic modifiers of the human immune response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.30.542928v1?rss=1">
<title>
<![CDATA[
MFGE8 inhibits insulin signaling through PTP1B 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.30.542928v1?rss=1</link>
<description><![CDATA[
The role of integrins in regulating insulin signaling is incompletely understood. We have previously shown that binding of the integrin ligand milk fat globule epidermal growth factor like 8 (MFGE8) to the v{beta}5 integrin promotes termination of insulin receptor signaling in mice. Upon ligation of MFGE8, {beta}5 complexes with the insulin receptor beta (IR{beta}) in skeletal muscle resulting in dephosphorylation of IR{beta} and reduction of insulin-stimulated glucose uptake. Here we investigate the mechanism by which the interaction between {beta}5 and IR{beta} impacts IR{beta} phosphorylation status. We show that {beta}5 blockade inhibits and MFGE8 promotes PTP1B binding to and dephosphorylation of IR{beta} resulting in reduced or increased insulin-stimulated myotube glucose uptake respectively. The {beta}5-PTP1B complex is recruited by MFGE8 to IR{beta} leading to termination of canonical insulin signaling. {beta}5 blockade enhances insulin-stimulated glucose uptake in wild type but not Ptp1b KO mice indicating that PTP1B functions downstream of MFGE8 in modulating insulin receptor signaling. Furthermore, in a human cohort, we report serum MFGE8 levels correlate with indices of insulin resistance. These data provide mechanistic insights into the role of MFGE8 and {beta}5 in regulating insulin signaling.
]]></description>
<dc:creator>Datta, R.</dc:creator>
<dc:creator>Podolsky, M. J.</dc:creator>
<dc:creator>Yang, C. D.</dc:creator>
<dc:creator>Alba, D.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Koliwad, S.</dc:creator>
<dc:creator>Lizama, C. O.</dc:creator>
<dc:creator>Atabai, K.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.05.30.542928</dc:identifier>
<dc:title><![CDATA[MFGE8 inhibits insulin signaling through PTP1B]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.31.543114v1?rss=1">
<title>
<![CDATA[
Evolution of the mutation spectrum across a mammalian phylogeny 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.31.543114v1?rss=1</link>
<description><![CDATA[
Little is known about how the spectrum and etiology of germline mutagenesis might vary among mammalian species. To shed light on this mystery, we quantify variation in mutational sequence context biases using polymorphism data from thirteen species of mice, apes, bears, wolves, and cetaceans. After normalizing the mutation spectrum for reference genome accessibility and k-mer content, we use the Mantel test to deduce that mutation spectrum divergence is highly correlated with genetic divergence between species, whereas life history traits like reproductive age are weaker predictors of mutation spectrum divergence. Potential bioinformatic confounders are only weakly related to a small set of mutation spectrum features. We find that clocklike mutational signatures previously inferred from human cancers cannot explain the phylogenetic signal exhibited by the mammalian mutation spectrum, despite the ability of these clocklike signatures to fit each species 3-mer spectrum with high cosine similarity. In contrast, parental aging signatures inferred from human de novo mutation data appear to explain much of the mutation spectrums phylogenetic signal when fit to non-context-dependent mutation spectrum data in combination with a novel mutational signature. We posit that future models purporting to explain the etiology of mammalian mutagenesis need to capture the fact that more closely related species have more similar mutation spectra; a model that fits each marginal spectrum with high cosine similarity is not guaranteed to capture this hierarchy of mutation spectrum variation among species.
]]></description>
<dc:creator>Beichman, A. C.</dc:creator>
<dc:creator>Robinson, J. A.</dc:creator>
<dc:creator>Lin, M.</dc:creator>
<dc:creator>Moreno-Estrada, A.</dc:creator>
<dc:creator>Nigenda-Morales, S.</dc:creator>
<dc:creator>Harris, K.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.05.31.543114</dc:identifier>
<dc:title><![CDATA[Evolution of the mutation spectrum across a mammalian phylogeny]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.01.543220v1?rss=1">
<title>
<![CDATA[
Phage anti-CBASS protein simultaneously sequesters cyclic trinucleotides and dinucleotides 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.01.543220v1?rss=1</link>
<description><![CDATA[
CBASS is a common anti-phage immune system that uses cyclic oligonucleotide signals to activate effectors and limit phage replication. In turn, phages encode anti-CBASS (Acb) proteins. We recently uncovered a widespread phage anti-CBASS protein Acb2 that acts as a "sponge" by forming a hexamer complex with three cGAMP molecules. Here, we identified that Acb2 binds and sequesters many CBASS and cGAS-produced cyclic dinucleotides in vitro and inhibits cGAMP-mediated STING activity in human cells. Surprisingly, Acb2 also binds CBASS cyclic trinucleotides 333-cyclic AMP-AMP-AMP (cA3) and 333-cAAG with high affinity. Structural characterization identified a distinct binding pocket within the Acb2 hexamer that binds two cyclic trinucleotide molecules and another binding pocket that binds to cyclic dinucleotides. Binding in one pocket does not allosterically alter the other, such that one Acb2 hexamer can simultaneously bind two cyclic trinucleotides and three cyclic dinucleotides. Phage-encoded Acb2 provides protection from Type III-C CBASS that uses cA3 signaling molecules in vivo and blocks cA3-mediated activation of the endonuclease effector in vitro. Altogether, Acb2 sequesters nearly all known CBASS signaling molecules through two distinct binding pockets and therefore serves as a broad-spectrum inhibitor of cGAS-based immunity.
]]></description>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Xiao, Y.</dc:creator>
<dc:creator>Huiting, E.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Ren, J.</dc:creator>
<dc:creator>Guan, L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Feng, Y.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.06.01.543220</dc:identifier>
<dc:title><![CDATA[Phage anti-CBASS protein simultaneously sequesters cyclic trinucleotides and dinucleotides]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.30.542978v1?rss=1">
<title>
<![CDATA[
Deep learning from harmonized peptide libraries enables retention time prediction of diverse post translational modifications 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.30.542978v1?rss=1</link>
<description><![CDATA[
In proteomics experiments, peptide retention time (RT) is an orthogonal property to fragmentation when assessing detection confidence. Advances in deep learning enable accurate RT prediction for any peptide from sequence alone, including those yet to be experimentally observed. Here we present Chronologer, an open-source software tool for rapid and accurate peptide RT prediction. Using new approaches to harmonize and false-discovery correct across independently collected datasets, Chronologer is built on a massive database with >2.2 million peptides including 10 common post-translational modification (PTM) types. By linking knowledge learned across diverse peptide chemistries, Chronologer predicts RTs with less than two-thirds the error of other deep learning tools. We show how RT for rare PTMs, such as OGlcNAc, can be learned with high accuracy using as few as 10-100 example peptides in newly harmonized datasets. This iteratively updatable workflow enables Chronologer to comprehensively predict RTs for PTM-marked peptides across entire proteomes.
]]></description>
<dc:creator>Wilburn, D. B.</dc:creator>
<dc:creator>Shannon, A. E.</dc:creator>
<dc:creator>Spicer, V.</dc:creator>
<dc:creator>Richards, A. L.</dc:creator>
<dc:creator>Yeung, D.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Krokhin, O. V.</dc:creator>
<dc:creator>Searle, B. C.</dc:creator>
<dc:date>2023-06-02</dc:date>
<dc:identifier>doi:10.1101/2023.05.30.542978</dc:identifier>
<dc:title><![CDATA[Deep learning from harmonized peptide libraries enables retention time prediction of diverse post translational modifications]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.03.542675v1?rss=1">
<title>
<![CDATA[
Structural characterization of ligand binding and pH-specific enzymatic activity of mouse Acidic Mammalian Chitinase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.03.542675v1?rss=1</link>
<description><![CDATA[
Chitin is an abundant biopolymer and pathogen-associated molecular pattern that stimulates a host innate immune response. Mammals express chitin-binding and chitin-degrading proteins to remove chitin from the body. One of these proteins, Acidic Mammalian Chitinase (AMCase), is an enzyme known for its ability to function under acidic conditions in the stomach but is also active in tissues with more neutral pHs, such as the lung. Here, we used a combination of biochemical, structural, and computational modeling approaches to examine how the mouse homolog (mAMCase) can act in both acidic and neutral environments. We measured kinetic properties of mAMCase activity across a broad pH range, quantifying its unusual dual activity optima at pH 2 and 7. We also solved high resolution crystal structures of mAMCase in complex with oligomeric GlcNAcn, the building block of chitin, where we identified extensive conformational ligand heterogeneity. Leveraging these data, we conducted molecular dynamics simulations that suggest how a key catalytic residue could be protonated via distinct mechanisms in each of the two environmental pH ranges. These results integrate structural, biochemical, and computational approaches to deliver a more complete understanding of the catalytic mechanism governing mAMCase activity at different pH. Engineering proteins with tunable pH optima may provide new opportunities to develop improved enzyme variants, including AMCase, for therapeutic purposes in chitin degradation.
]]></description>
<dc:creator>Diaz, R. E.</dc:creator>
<dc:creator>Ecker, A. K.</dc:creator>
<dc:creator>Correy, G. J.</dc:creator>
<dc:creator>Asthana, P.</dc:creator>
<dc:creator>Young, I. D.</dc:creator>
<dc:creator>Faust, B.</dc:creator>
<dc:creator>Thompson, M. C.</dc:creator>
<dc:creator>Seiple, I. B.</dc:creator>
<dc:creator>Van Dyken, S. J.</dc:creator>
<dc:creator>Locksley, R. M.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2023-06-03</dc:date>
<dc:identifier>doi:10.1101/2023.06.03.542675</dc:identifier>
<dc:title><![CDATA[Structural characterization of ligand binding and pH-specific enzymatic activity of mouse Acidic Mammalian Chitinase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.02.543290v1?rss=1">
<title>
<![CDATA[
DNAJB1-PRKACA fusion protein-regulated LINC00473 promotes tumor growth and alters mitochondrial fitness in fibrolamellar carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.02.543290v1?rss=1</link>
<description><![CDATA[
Fibrolamellar carcinoma (FLC) is a rare liver cancer that disproportionately affects adolescents and young adults. Currently, no standard of care is available and there remains a dire need for new therapeutics. Most patients harbor the fusion oncogene DNAJB1-PRKACA (DP fusion), but clinical inhibitors are not yet developed and it is critical to identify downstream mediators of FLC pathogenesis. Here, we identify long non-coding RNA LINC00473 among the most highly upregulated genes in FLC tumors and determine that it is strongly suppressed by RNAi-mediated inhibition of the DP fusion in FLC tumor epithelial cells. We show by loss- and gain-of-function studies that LINC00473 suppresses apoptosis, increases the expression of FLC marker genes, and promotes FLC growth in cell-based and in vivo models of disease. Mechanistically, LINC00473 plays an important role in promoting glycolysis and altering mitochondrial activity. Specifically, LINC00473 knockdown leads to increased spare respiratory capacity, an indicator of mitochondrial fitness. Overall, we propose that LINC00473 could be a viable target for this devastating disease.

HighlightsFibrolamellar carcinoma (FLC) is a lethal liver cancer lacking effective therapeutic options. Ma et al. demonstrate that primate-specific RNA LINC00473 is enriched in tumor epithelial cells and functions to promote FLC growth and dysregulate cellular energetics, unveiling an important mechanism downstream of the fusion oncogene, DNAJB1-PRKACA, in FLC pathogenesis.

In BriefO_LILINC00473 is consistently elevated in primary FLC tumor tissue from different patient cohorts and in multiple disease models.
C_LIO_LIDP fusion, the signature oncoprotein of FLC, drives LINC00473 expression.
C_LIO_LILINC00473 promotes FLC growth via anti-apoptotic function.
C_LIO_LILINC00473 modulates FLC energetics by promoting glycolysis and altering mitochondrial fitness.
C_LI



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/543290v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Ma, R. K.</dc:creator>
<dc:creator>Tsai, P.-Y.</dc:creator>
<dc:creator>Farghli, A. R.</dc:creator>
<dc:creator>Shumway, A. J.</dc:creator>
<dc:creator>Kanke, M.</dc:creator>
<dc:creator>Gordan, J. D.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:creator>Vakili, K.</dc:creator>
<dc:creator>Nukaya, M.</dc:creator>
<dc:creator>Noetzli, L.</dc:creator>
<dc:creator>Ronnekleiv-Kelly, S.</dc:creator>
<dc:creator>Broom, W.</dc:creator>
<dc:creator>Barrow, J.</dc:creator>
<dc:creator>Sethupathy, P.</dc:creator>
<dc:date>2023-06-04</dc:date>
<dc:identifier>doi:10.1101/2023.06.02.543290</dc:identifier>
<dc:title><![CDATA[DNAJB1-PRKACA fusion protein-regulated LINC00473 promotes tumor growth and alters mitochondrial fitness in fibrolamellar carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.02.543391v1?rss=1">
<title>
<![CDATA[
Mechanism of substrate binding and transport in BASS transporters 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.02.543391v1?rss=1</link>
<description><![CDATA[
The Bile Acid Sodium Symporter (BASS) family transports a wide array of molecules across membranes, including bile acids in humans, and small metabolites in plants. These transporters, many of which are sodium-coupled, have been shown to use an elevator mechanism of transport, but exactly how substrate binding is coupled to sodium ion binding and transport is not clear. Here we solve the crystal structure at 2.3 [A] of a transporter from Neisseria Meningitidis (ASBTNM) in complex with pantoate, a potential substrate of ASBTNM. The BASS family is characterised by two helices that cross-over in the centre of the protein in an arrangement that is intricately held together by two sodium ions. We observe that the pantoate binds, specifically, between the N-termini of two of the opposing helices in this cross-over region. During molecular dynamics simulations the pantoate remains in this position when sodium ions are present but is more mobile in their absence. Comparison of structures in the presence and absence of pantoate demonstrates that pantoate elicits a conformational change in one of the cross-over helices. This modifies the interface between the two domains that move relative to one another to elicit the elevator mechanism. These results have implications, not only for ASBTNM but for the BASS family as a whole and indeed other transporters that work through the elevator mechanism.
]]></description>
<dc:creator>Becker, P.</dc:creator>
<dc:creator>Naughton, F. B.</dc:creator>
<dc:creator>Brotherton, D. H.</dc:creator>
<dc:creator>Pacheco-Gomez, R.</dc:creator>
<dc:creator>Beckstein, O.</dc:creator>
<dc:creator>Cameron, A. D.</dc:creator>
<dc:date>2023-06-04</dc:date>
<dc:identifier>doi:10.1101/2023.06.02.543391</dc:identifier>
<dc:title><![CDATA[Mechanism of substrate binding and transport in BASS transporters]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.01.543325v1?rss=1">
<title>
<![CDATA[
A novel small molecule PKC epsilon inhibitor reduces hyperalgesia induced by paclitaxel or opioid withdrawal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.01.543325v1?rss=1</link>
<description><![CDATA[
Aggressive marketing and increased prescribing of opioids for treating pain have fueled a prominent increase in opioid use disorder. This acute public health problem has led to calls for the development of non-opioid alternatives to treat chronic pain. The enzyme protein kinase C epsilon (PKC{varepsilon}) plays an important role in nociceptor sensitization, in inflammatory and neuropathic pain. Here we investigated the effects of a novel small molecule that inhibits PKC{varepsilon} in a rodent model of chemotherapy-induced neuropathic pain produced by administration of the cancer chemotherapy, paclitaxel. Because transition of opioid-dependent individuals to a non-opioid pain medication can increase pain due to opioid withdrawal, we also investigated if the PKC{varepsilon} inhibitor alters features of opioid withdrawal and opioid self-administration. This novel PKC{varepsilon} inhibitor attenuated paclitaxel-induced hyperalgesia, reversed hyperalgesia produced by opioid withdrawal, and reduced somatic signs of opioid withdrawal. This PKC{varepsilon} inhibitor did not modify opioid self-administration, nor produce self-administration. These findings suggest that PKC{varepsilon} inhibition is an effective, non-addictive strategy to treat chemotherapy-induced neuropathic pain with the added benefit of limiting hyperalgesia due to opioid withdrawal, which could facilitate switching treatment of chronic pain, in opioid-dependent individuals.
]]></description>
<dc:creator>Gregory-Flores, A.</dc:creator>
<dc:creator>Magayewski Bonet, I. J.</dc:creator>
<dc:creator>Desaivre, S.</dc:creator>
<dc:creator>Levine, J. D.</dc:creator>
<dc:creator>McHardy, S. F.</dc:creator>
<dc:creator>de Kraker, H.</dc:creator>
<dc:creator>Russell, N.</dc:creator>
<dc:creator>Fleischer, C.</dc:creator>
<dc:creator>Messing, R.</dc:creator>
<dc:creator>Marinelli, M.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.01.543325</dc:identifier>
<dc:title><![CDATA[A novel small molecule PKC epsilon inhibitor reduces hyperalgesia induced by paclitaxel or opioid withdrawal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.05.543690v1?rss=1">
<title>
<![CDATA[
Mesostriatal dopamine is sensitive to specific cue-reward contingencies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.05.543690v1?rss=1</link>
<description><![CDATA[
Learning causal relationships relies on understanding how often one event precedes another. To gain an understanding of how dopamine neuron activity and neurotransmitter release change when a retrospective relationship is degraded for a specific pair of events, we used outcome-selective Pavlovian contingency degradation in rats. Two cues were paired with distinct food rewards, one of which was also delivered in the absence of either cue. Conditioned approach was attenuated for the cue-reward contingency that was degraded. Dopamine neuron activity in the midbrain and dopamine release in the ventral striatum showed a profile of changes in cue- and reward-evoked responding that was not easily explained by a standard reinforcement learning model. An alternative model based on learning causal relationships was better able to capture evoked dopamine responses during contingency degradation, as well as conditioned behavior following optogenetic manipulations of dopamine during noncontingent rewards. Our results suggest that mesostriatal dopamine encodes the contingencies between meaningful events during learning.
]]></description>
<dc:creator>Garr, E.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Brooke, S.</dc:creator>
<dc:creator>Castell, L.</dc:creator>
<dc:creator>Bal, A.</dc:creator>
<dc:creator>Magnard, R.</dc:creator>
<dc:creator>Namboodiri, V. M. K.</dc:creator>
<dc:creator>Janak, P. H.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.05.543690</dc:identifier>
<dc:title><![CDATA[Mesostriatal dopamine is sensitive to specific cue-reward contingencies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.05.543428v1?rss=1">
<title>
<![CDATA[
NEMO reshapes the protein aggregate interface and promotes aggrephagy by co-condensation with p62 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.05.543428v1?rss=1</link>
<description><![CDATA[
NEMO is a ubiquitin-binding protein which regulates canonical NF-{kappa}B pathway activation in innate immune signaling, cell death regulation and host-pathogen interactions. Here we identified an NF-{kappa}B-independent function of NEMO in proteostasis regulation by promoting autophagosomal clearance of protein aggregates. NEMO-deficient cells accumulate misfolded proteins upon proteotoxic stress and are vulnerable to proteostasis challenges. Moreover, a patient with a mutation in the NEMO gene resulting in defective binding of NEMO to linear ubiquitin chains, developed a widespread mixed brain proteinopathy, including -synuclein, tau and TDP-43 pathology. NEMO amplifies linear ubiquitylation at -synuclein aggregates and promotes the local concentration of p62 into foci. In vitro, NEMO lowers the threshold concentrations required for ubiquitin-dependent phase transition of p62. In summary, NEMO reshapes the aggregate surface for efficient autophagosomal clearance by providing a mobile phase at the aggregate interphase favoring co-condensation with p62.
]]></description>
<dc:creator>Furthmann, N.</dc:creator>
<dc:creator>Angersbach, L.</dc:creator>
<dc:creator>Bader, V.</dc:creator>
<dc:creator>Blusch, A.</dc:creator>
<dc:creator>Goel, S.</dc:creator>
<dc:creator>Sanchez-Vicente, A.</dc:creator>
<dc:creator>Krause, L. J.</dc:creator>
<dc:creator>Grover, P.</dc:creator>
<dc:creator>Trinkaus, V. A.</dc:creator>
<dc:creator>van Well, E. M.</dc:creator>
<dc:creator>Jaugstetter, M.</dc:creator>
<dc:creator>Tschulik, K.</dc:creator>
<dc:creator>Damgaard, R. B.</dc:creator>
<dc:creator>Saft, C.</dc:creator>
<dc:creator>Ellrichmann, G.</dc:creator>
<dc:creator>Gold, R.</dc:creator>
<dc:creator>Koch, A.</dc:creator>
<dc:creator>Englert, B.</dc:creator>
<dc:creator>Glatzel, M.</dc:creator>
<dc:creator>Hartl, F.-U.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Christine, C.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Tatzelt, J.</dc:creator>
<dc:creator>Winklhofer, K. F.</dc:creator>
<dc:date>2023-06-06</dc:date>
<dc:identifier>doi:10.1101/2023.06.05.543428</dc:identifier>
<dc:title><![CDATA[NEMO reshapes the protein aggregate interface and promotes aggrephagy by co-condensation with p62]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.05.543758v1?rss=1">
<title>
<![CDATA[
Post-acute immunological and behavioral sequelae in mice after Omicron infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.05.543758v1?rss=1</link>
<description><![CDATA[
Progress in understanding long COVID and developing effective therapeutics is hampered in part by the lack of suitable animal models. Here we used ACE2-transgenic mice recovered from Omicron (BA.1) infection to test for pulmonary and behavioral post-acute sequelae. Through in-depth phenotyping by CyTOF, we demonstrate that naive mice experiencing a first Omicron infection exhibit profound immune perturbations in the lung after resolving acute infection. This is not observed if mice were first vaccinated with spike-encoding mRNA. The protective effects of vaccination against post-acute sequelae were associated with a highly polyfunctional SARS-CoV-2-specific T cell response that was recalled upon BA.1 breakthrough infection but not seen with BA.1 infection alone. Without vaccination, the chemokine receptor CXCR4 was uniquely upregulated on multiple pulmonary immune subsets in the BA.1 convalescent mice, a process previously connected to severe COVID-19. Taking advantage of recent developments in machine learning and computer vision, we demonstrate that BA.1 convalescent mice exhibited spontaneous behavioral changes, emotional alterations, and cognitive-related deficits in context habituation. Collectively, our data identify immunological and behavioral post-acute sequelae after Omicron infection and uncover a protective effect of vaccination against post-acute pulmonary immune perturbations.
]]></description>
<dc:creator>Ma, T.</dc:creator>
<dc:creator>Suryawanshi, R. K.</dc:creator>
<dc:creator>Miller, S. R.</dc:creator>
<dc:creator>Ly, K. K.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Elphick, N.</dc:creator>
<dc:creator>Yin, K.</dc:creator>
<dc:creator>Luo, X.</dc:creator>
<dc:creator>Kaliss, N.</dc:creator>
<dc:creator>Chen, I. P.</dc:creator>
<dc:creator>Montano, M.</dc:creator>
<dc:creator>Sreekumar, B.</dc:creator>
<dc:creator>Standker, L.</dc:creator>
<dc:creator>Munch, J.</dc:creator>
<dc:creator>Damron, F. H.</dc:creator>
<dc:creator>Palop, J. J.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Roan, N. R.</dc:creator>
<dc:date>2023-06-07</dc:date>
<dc:identifier>doi:10.1101/2023.06.05.543758</dc:identifier>
<dc:title><![CDATA[Post-acute immunological and behavioral sequelae in mice after Omicron infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.05.543814v1?rss=1">
<title>
<![CDATA[
Impaired cerebellar plasticity hypersensitizessensory reflexes in SCN2A-associated ASD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.05.543814v1?rss=1</link>
<description><![CDATA[
Children diagnosed with autism spectrum disorder (ASD) commonly present with sensory hypersensitivity, or abnormally strong reactions to sensory stimuli. Such hypersensitivity can be overwhelming, causing high levels of distress that contribute markedly to the negative aspects of the disorder. Here, we identify the mechanisms that underlie hypersensitivity in a sensorimotor reflex found to be altered in humans and in mice with loss-of-function in the ASD risk-factor gene SCN2A. The cerebellum-dependent vestibulo-ocular reflex (VOR), which helps maintain ones gaze during movement, was hypersensitized due to deficits in cerebellar synaptic plasticity. Heterozygous loss of SCN2A-encoded NaV1.2 sodium channels in granule cells impaired high-frequency transmission to Purkinje cells and long-term potentiation, a form of synaptic plasticity important for modulating VOR gain. VOR plasticity could be rescued in adolescent mice via a CRISPR-activator approach that increases Scn2a expression, highlighting how evaluation of simple reflexes can be used as quantitative readout of therapeutic interventions.
]]></description>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Derderian, K. D.</dc:creator>
<dc:creator>Hamada, E.</dc:creator>
<dc:creator>Zhou, X.</dc:creator>
<dc:creator>Nelson, A. D.</dc:creator>
<dc:creator>Kyoung, H.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Bouvier, G.</dc:creator>
<dc:creator>Bender, K.</dc:creator>
<dc:date>2023-06-07</dc:date>
<dc:identifier>doi:10.1101/2023.06.05.543814</dc:identifier>
<dc:title><![CDATA[Impaired cerebellar plasticity hypersensitizessensory reflexes in SCN2A-associated ASD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.06.543768v1?rss=1">
<title>
<![CDATA[
The Neonatal Gyrencephalic Cortex Maintains Regionally Distinct Streams of Neuroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.06.543768v1?rss=1</link>
<description><![CDATA[
Neurodevelopmental mechanisms have evolved to support the formation of diverse brain structures, such as in humans, during the perinatal period. Here, we demonstrate that neonatal gyrencephalic brains harbor an expanded subventricular zone, termed the Arc, defined by tiered arrangement of doublecortin (DCX)-expressing neuroblasts and vascular enrichment at the ventricular wall. The Arc is the origin of dorsal and ventral populations of migratory neuroblasts that target multiple regions involved in higher cognitive functions. Arc-derived migratory streams, primarily from the caudal ganglionic eminence, are composed of diverse neuronal subtypes with distinct spatial and migratory-receptor profiles. Our findings indicate the Arc is a structure present in phylogenetically divergent species that supports the expansion of postnatal neuronal migration, contributing to a protracted formation of cortical circuits in gyrencephalic brains.

One-Sentence SummaryThe ventricular cytoarchitecture of gyrencephalic brains supports an ongoing supply of migratory neurons to the neonatal cortex.
]]></description>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Sandoval, K.</dc:creator>
<dc:creator>Poddar, A.</dc:creator>
<dc:creator>Chu, J.</dc:creator>
<dc:creator>Horton, E.</dc:creator>
<dc:creator>Cui, D.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Bartels, T.</dc:creator>
<dc:creator>Wood, C.</dc:creator>
<dc:creator>Rowitch, D. H.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Sherwood, C. C.</dc:creator>
<dc:creator>Kramer, B. W.</dc:creator>
<dc:creator>Roberts, A. C.</dc:creator>
<dc:creator>ROSS, P. J.</dc:creator>
<dc:creator>Xu, D.</dc:creator>
<dc:creator>Robertson, N. J.</dc:creator>
<dc:creator>Ji, P.</dc:creator>
<dc:creator>Maga, E. A.</dc:creator>
<dc:creator>Paredes, M. F.</dc:creator>
<dc:date>2023-06-07</dc:date>
<dc:identifier>doi:10.1101/2023.06.06.543768</dc:identifier>
<dc:title><![CDATA[The Neonatal Gyrencephalic Cortex Maintains Regionally Distinct Streams of Neuroblasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.07.544129v1?rss=1">
<title>
<![CDATA[
Quantifying turgor pressure in budding and fission yeasts based upon osmotic properties 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.07.544129v1?rss=1</link>
<description><![CDATA[
Walled cells, such as plants, fungi, and bacteria cells, possess a high internal hydrostatic pressure, termed turgor pressure, that drives volume growth and contributes to cell shape determination. Rigorous measurement of turgor pressure, however, remains challenging, and reliable quantitative measurements, even in budding yeast are still lacking. Here, we present a simple and robust experimental approach to access turgor pressure in yeasts based upon the determination of isotonic concentration using protoplasts as osmometers. We propose three methods to identify the isotonic condition - 3D cell volume, cytoplasmic fluorophore intensity, and mobility of a cytGEMs nano-rheology probe - that all yield consistent values. Our results provide turgor pressure estimates of 1.0 {+/-} 0.1 MPa for S. pombe, 0.49 {+/-} 0.01 MPa for S. japonicus, 0.5 {+/-} 0.1 MPa for S. cerevisiae W303a and 0.31 {+/-} 0.03 MPa for S. cerevisiae BY4741. Large differences in turgor pressure and nano-rheology measurements between the S. cerevisiae strains demonstrate how fundamental biophysical parameters can vary even among wildtype strains of the same species. These side-by-side measurements of turgor pressure in multiple yeast species provide critical values for quantitative studies on cellular mechanics and comparative evolution.
]]></description>
<dc:creator>LEMIERE, J.</dc:creator>
<dc:creator>Chang, F.</dc:creator>
<dc:date>2023-06-07</dc:date>
<dc:identifier>doi:10.1101/2023.06.07.544129</dc:identifier>
<dc:title><![CDATA[Quantifying turgor pressure in budding and fission yeasts based upon osmotic properties]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.06.543909v1?rss=1">
<title>
<![CDATA[
Neuronal Activation of the Gαq Protein EGL-30/GNAQ Late in Life Rejuvenates Cognition Across Species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.06.543909v1?rss=1</link>
<description><![CDATA[
Cognitive decline is perhaps the most devastating aging loss. EGL-30/GNAQ and Gq signaling pathways are highly conserved between C. elegans and mammals. We find that activation of EGL-30 in aged worms at least triples memory span, and we wondered if this highly conserved pathway could also improve memory in very old mice. Murine Gnaq is enriched in hippocampal excitatory neurons and declines with age. Furthermore, GNAQ gain-of-function significantly improved memory in aged mice: GNAQ(gf) in hippocampal neurons of 24-month-old mice rescued age-related impairments in health metrics and long-term memory. Single-nucleus RNAseq revealed gene expression changes related to synaptic function, axon guidance, and learning and memory pathways. Several worm orthologs of mouse genes upregulated by GNAQ(gf) overexpression are required for EGL-30(gf)-dependent memory improvement. These results demonstrate that the molecular and genetic pathways between C. elegans and mammals are highly conserved, as activation of EGL-30/GNAQ, a pathway first identified in worms, rejuvenates cognitive function in two-year old mice (the equivalent of 70-80 yo humans). To our knowledge, this is the oldest age an intervention has successfully improved age-related cognitive decline.

One-Sentence SummaryNeuronal activation of the Gq protein EGL-30/GNAQ restores long-term memory at old age in worms and mice.
]]></description>
<dc:creator>Stevenson, M. E.</dc:creator>
<dc:creator>Bieri, G.</dc:creator>
<dc:creator>Kaletsky, R.</dc:creator>
<dc:creator>Ange, J. S.</dc:creator>
<dc:creator>Remesal, L.</dc:creator>
<dc:creator>Pratt, K. J. B.</dc:creator>
<dc:creator>Zhou, S.</dc:creator>
<dc:creator>Weng, Y.</dc:creator>
<dc:creator>Murphy, C. T.</dc:creator>
<dc:creator>Villeda, S. A.</dc:creator>
<dc:date>2023-06-11</dc:date>
<dc:identifier>doi:10.1101/2023.06.06.543909</dc:identifier>
<dc:title><![CDATA[Neuronal Activation of the Gαq Protein EGL-30/GNAQ Late in Life Rejuvenates Cognition Across Species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.09.544373v1?rss=1">
<title>
<![CDATA[
Neuropathy target esterase activity predicts retinopathy among PNPLA6 disorders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.09.544373v1?rss=1</link>
<description><![CDATA[
Biallelic pathogenic variants in the PNPLA6 gene cause a broad spectrum of disorders leading to gait disturbance, visual impairment, anterior hypopituitarism, and hair anomalies. PNPLA6 encodes Neuropathy target esterase (NTE), yet the role of NTE dysfunction on affected tissues in the large spectrum of associated disease remains unclear. We present a clinical meta-analysis of a novel cohort of 23 new patients along with 95 reported individuals with PNPLA6 variants that implicate missense variants as a driver of disease pathogenesis. Measuring esterase activity of 46 disease-associated and 20 common variants observed across PNPLA6-associated clinical diagnoses unambiguously reclassified 10 variants as likely pathogenic and 36 variants as pathogenic, establishing a robust functional assay for classifying PNPLA6 variants of unknown significance. Estimating the overall NTE activity of affected individuals revealed a striking inverse relationship between NTE activity and the presence of retinopathy and endocrinopathy. This phenomenon was recaptured in vivo in an allelic mouse series, where a similar NTE threshold for retinopathy exists. Thus, PNPLA6 disorders, previously considered allelic, are a continuous spectrum of pleiotropic phenotypes defined by an NTE genotype:activity:phenotype relationship. This relationship and the generation of a preclinical animal model pave the way for therapeutic trials, using NTE as a biomarker.
]]></description>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Lwin, C.</dc:creator>
<dc:creator>Han, M.</dc:creator>
<dc:creator>Guan, B.</dc:creator>
<dc:creator>Niak, A.</dc:creator>
<dc:creator>Bender, C.</dc:creator>
<dc:creator>Moore, N.</dc:creator>
<dc:creator>Huryn, L. A.</dc:creator>
<dc:creator>Sergeev, Y.</dc:creator>
<dc:creator>Qian, H.</dc:creator>
<dc:creator>Zeng, Y.</dc:creator>
<dc:creator>Dong, L.</dc:creator>
<dc:creator>Liu, P.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Haugen, C. J.</dc:creator>
<dc:creator>Prasov, L.</dc:creator>
<dc:creator>Shi, R.</dc:creator>
<dc:creator>Dollfus, H.</dc:creator>
<dc:creator>Aristodemou, P.</dc:creator>
<dc:creator>Laich, Y.</dc:creator>
<dc:creator>Nemeth, A. H.</dc:creator>
<dc:creator>Taylor, J.</dc:creator>
<dc:creator>Downes, S.</dc:creator>
<dc:creator>Krawczynski, M.</dc:creator>
<dc:creator>Meunier, I.</dc:creator>
<dc:creator>Strassberg, M.</dc:creator>
<dc:creator>Tenney, J.</dc:creator>
<dc:creator>Gao, J.</dc:creator>
<dc:creator>Shear, M. A.</dc:creator>
<dc:creator>Moore, A. T.</dc:creator>
<dc:creator>Duncan, J. L.</dc:creator>
<dc:creator>Menendez, B.</dc:creator>
<dc:creator>Hull, S.</dc:creator>
<dc:creator>Vincent, A.</dc:creator>
<dc:creator>Siskind, C. E.</dc:creator>
<dc:creator>Traboulsi, E. I.</dc:creator>
<dc:creator>Blackstone, C.</dc:creator>
<dc:creator>Sisk, R.</dc:creator>
<dc:creator>Utz, V.</dc:creator>
<dc:creator>Webster, A. R.</dc:creator>
<dc:creator>Michaelides, M.</dc:creator>
<dc:creator>Arno, G.</dc:creator>
<dc:creator>Synofzik, M.</dc:creator>
<dc:creator>Hufnagel, R. B.</dc:creator>
<dc:date>2023-06-11</dc:date>
<dc:identifier>doi:10.1101/2023.06.09.544373</dc:identifier>
<dc:title><![CDATA[Neuropathy target esterase activity predicts retinopathy among PNPLA6 disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.09.544427v1?rss=1">
<title>
<![CDATA[
Homologous chromosome recognition via nonspecific interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.09.544427v1?rss=1</link>
<description><![CDATA[
In many organisms, most notably Drosophila, homologous chromosomes in somatic cells associate with each other, a phenomenon known as somatic homolog pairing. Unlike in meiosis, where homology is read out at the level of DNA sequence complementarity, somatic homolog pairing takes place without double strand breaks or strand invasion, thus requiring some other mechanism for homologs to recognize each other. Several studies have suggested a "specific button" model, in which a series of distinct regions in the genome, known as buttons, can associate with each other, presumably mediated by different proteins that bind to these different regions. Here we consider an alternative model, which we term the "button barcode" model, in which there is only one type of recognition site or adhesion button, present in many copies in the genome, each of which can associate with any of the others with equal affinity. An important component of this model is that the buttons are non-uniformly distributed, such that alignment of a chromosome with its correct homolog, compared with a non-homolog, is energetically favored; since to achieve nonhomologous alignment, chromosomes would be required to mechanically deform in order to bring their buttons into mutual register. We investigated several types of barcodes and examined their effect on pairing fidelity. We found that high fidelity homolog recognition can be achieved by arranging chromosome pairing buttons according to an actual industrial barcode used for warehouse sorting. By simulating randomly generated non-uniform button distributions, many highly effective button barcodes can be easily found, some of which achieve virtually perfect pairing fidelity. This model is consistent with existing literature on the effect of translocations of different sizes on homolog pairing. We conclude that a button barcode model can attain highly specific homolog recognition, comparable to that seen in actual cells undergoing somatic homolog pairing, without the need for specific interactions. This model may have implications for how meiotic pairing is achieved.
]]></description>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:creator>Fung, J. C.</dc:creator>
<dc:date>2023-06-11</dc:date>
<dc:identifier>doi:10.1101/2023.06.09.544427</dc:identifier>
<dc:title><![CDATA[Homologous chromosome recognition via nonspecific interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.12.543777v1?rss=1">
<title>
<![CDATA[
KIBRA repairs synaptic plasticity and promotes resilience to tauopathy-related memory loss 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.12.543777v1?rss=1</link>
<description><![CDATA[
Synaptic plasticity is obstructed by pathogenic tau in the brain, representing a key mechanism that underlies memory loss in Alzheimers disease (AD) and related tauopathies. Here, we define a mechanism for plasticity repair in vulnerable neurons using the C-terminus of the KIdney/BRAin (KIBRA) protein (CT-KIBRA). We show that CT-KIBRA restores plasticity and memory in transgenic mice expressing pathogenic human tau; however, CT-KIBRA did not alter tau levels or prevent tau-induced synapse loss. Instead, we find that CT-KIBRA binds to and stabilizes protein kinase M{zeta} (PKM{zeta}) to maintain synaptic plasticity and memory despite tau mediated pathogenesis. In humans we find that reduced KIBRA in brain and increased KIBRA in cerebrospinal fluid are associated with cognitive impairment and pathological tau levels in disease. Thus, our results distinguish KIBRA both as a novel biomarker of synapse dysfunction in AD and as the foundation for a synapse repair mechanism to reverse cognitive impairment in tauopathy.
]]></description>
<dc:creator>Kauwe, G.</dc:creator>
<dc:creator>Pareja-Navarro, K.</dc:creator>
<dc:creator>Yao, L.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Wong, I.</dc:creator>
<dc:creator>Saloner, R.</dc:creator>
<dc:creator>Cifuentes, H.</dc:creator>
<dc:creator>Nana, A. L.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Le, D.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Sacktor, T.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:creator>Casaletto, K. B.</dc:creator>
<dc:creator>Tracy, T. E.</dc:creator>
<dc:date>2023-06-13</dc:date>
<dc:identifier>doi:10.1101/2023.06.12.543777</dc:identifier>
<dc:title><![CDATA[KIBRA repairs synaptic plasticity and promotes resilience to tauopathy-related memory loss]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.12.544677v1?rss=1">
<title>
<![CDATA[
Polarized Desmosome and Hemidesmosome Shedding via Exosomes is an Early Indicator of Outer Blood-Retina Barrier Dysfunction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.12.544677v1?rss=1</link>
<description><![CDATA[
The retinal pigmented epithelium (RPE) constitutes the outer blood-retinal barrier, enables photoreceptor function of the eye, and is constantly exposed to oxidative stress. As such, dysfunction of the RPE underlies pathology leading to development of age-related macular degeneration (AMD), the leading cause of vision loss among the elderly in industrialized nations. A major responsibility of the RPE is to process photoreceptor outer segments, which relies on the proper functioning of its endocytic pathways and endosomal trafficking. Exosomes and other extracellular vesicles from RPE are an essential part of these pathways and may be early indicators of cellular stress. To test the role of exosomes that may underlie the early stages of AMD, we used a polarized primary RPE cell culture model under chronic subtoxic oxidative stress. Unbiased proteomic analyses of highly purified basolateral exosomes from oxidatively stressed RPE cultures revealed changes in proteins involved in epithelial barrier integrity. There were also significant changes in proteins accumulating in the basal-side sub-RPE extracellular matrix during oxidative stress, that could be prevented with an inhibitor of exosome release. Thus, chronic subtoxic oxidative stress in primary RPE cultures induces changes in exosome content, including basal-side specific desmosome and hemidesmosome shedding via exosomes. These findings provide novel biomarkers of early cellular dysfunction and opportunity for therapeutic intervention in age-related retinal diseases, (e.g., AMD) and broadly from blood-CNS barriers in other neurodegenerative diseases.
]]></description>
<dc:creator>Hernandez, B.</dc:creator>
<dc:creator>Skiba, N. P.</dc:creator>
<dc:creator>Ploessl, K.</dc:creator>
<dc:creator>Strain, M.</dc:creator>
<dc:creator>Grigsby, D.</dc:creator>
<dc:creator>Kelly, U.</dc:creator>
<dc:creator>Cady, M. A.</dc:creator>
<dc:creator>Manocha, V.</dc:creator>
<dc:creator>Maminishkis, A.</dc:creator>
<dc:creator>Watkins, T.</dc:creator>
<dc:creator>Miller, S. S.</dc:creator>
<dc:creator>Ashley-Koch, A.</dc:creator>
<dc:creator>Stamer, W. D.</dc:creator>
<dc:creator>Weber, B. H.</dc:creator>
<dc:creator>Rickman, C. B.</dc:creator>
<dc:creator>Klingeborn, M.</dc:creator>
<dc:date>2023-06-13</dc:date>
<dc:identifier>doi:10.1101/2023.06.12.544677</dc:identifier>
<dc:title><![CDATA[Polarized Desmosome and Hemidesmosome Shedding via Exosomes is an Early Indicator of Outer Blood-Retina Barrier Dysfunction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.13.544831v1?rss=1">
<title>
<![CDATA[
Scalable, cell type-selective in vivo AAV-based CRISPR screens in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.13.544831v1?rss=1</link>
<description><![CDATA[
There is a significant need for scalable CRISPR-based genetic screening methods that can be applied directly in mammalian tissues in vivo while enabling cell type-specific analysis. To address this, we developed an adeno-associated virus (AAV)-based CRISPR screening platform, CrAAVe- seq, that incorporates a Cre-sensitive sgRNA construct for pooled screening within targeted cell populations in mouse tissues. We demonstrate the utility of this approach by screening two distinct large sgRNA libraries, together targeting over 5,000 genes, in mouse brains to create a robust profile of neuron-essential genes. We validate two genes as strongly neuron-essential in both primary mouse neurons and in vivo, confirming the predictive power of our platform. By comparing results from individual mice and across different cell populations, we highlight the reproducibility and scalability of the platform and show that it is highly sensitive even for screening smaller neuronal subpopulations. We systematically characterize the impact of sgRNA library size, mouse cohort size, the size of the targeted cell population, viral titer, and multiplicity of infection on screen performance to establish general guidelines for large-scale in vivo screens.
]]></description>
<dc:creator>Ramani, B.</dc:creator>
<dc:creator>Rose, I. V. L.</dc:creator>
<dc:creator>Pan, A.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Ma, K.</dc:creator>
<dc:creator>Palop, J. J.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:date>2023-06-13</dc:date>
<dc:identifier>doi:10.1101/2023.06.13.544831</dc:identifier>
<dc:title><![CDATA[Scalable, cell type-selective in vivo AAV-based CRISPR screens in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.13.544829v1?rss=1">
<title>
<![CDATA[
Intrinsic Homeostatic Plasticity in Mouse and Human Sensory Neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.13.544829v1?rss=1</link>
<description><![CDATA[
In response to changes in activity induced by environmental cues, neurons in the central nervous system undergo homeostatic plasticity to sustain overall network function during abrupt changes in synaptic strengths. Homeostatic plasticity involves changes in synaptic scaling and regulation of intrinsic excitability. Increases in spontaneous firing and excitability of sensory neurons are evident in some forms of chronic pain in animal models and human patients. However, whether mechanisms of homeostatic plasticity are engaged in sensory neurons under normal conditions or altered after chronic pain is unknown. Here, we showed that sustained depolarization induced by 30mM KCl induces a compensatory decrease in the excitability in mouse and human sensory neurons. Moreover, voltage-gated sodium currents are robustly reduced in mouse sensory neurons contributing to the overall decrease in neuronal excitability. Decreased efficacy of these homeostatic mechanisms could potentially contribute to the development of the pathophysiology of chronic pain.
]]></description>
<dc:creator>Mcllvried, L. A.</dc:creator>
<dc:creator>Del Rosario, J. S.</dc:creator>
<dc:creator>Pullen, M. Y.</dc:creator>
<dc:creator>Wangzhou, A.</dc:creator>
<dc:creator>Sheahan, T. D.</dc:creator>
<dc:creator>Shepherd, A. J.</dc:creator>
<dc:creator>Slivicki, R. A.</dc:creator>
<dc:creator>Lemen, J. A.</dc:creator>
<dc:creator>Price, T. J.</dc:creator>
<dc:creator>Copits, B. A.</dc:creator>
<dc:creator>Gereau, R. W.</dc:creator>
<dc:date>2023-06-14</dc:date>
<dc:identifier>doi:10.1101/2023.06.13.544829</dc:identifier>
<dc:title><![CDATA[Intrinsic Homeostatic Plasticity in Mouse and Human Sensory Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.14.545005v1?rss=1">
<title>
<![CDATA[
Hyperglycosylation of prosaposin in tumor DCs promotes immune escape in cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.14.545005v1?rss=1</link>
<description><![CDATA[
Tumors develop strategies to evade immunity by suppressing antigen presentation. Here, we show that prosaposin drives CD8 T cell-mediated tumor immunity and that its hyperglycosylation in tumor DCs leads to cancer immune escape. We found that lysosomal prosaposin and its single saposin cognates mediated disintegration of tumor cell-derived apoptotic bodies to facilitate presentation of membrane-associated antigen and T cell activation. In the tumor microenvironment, TGF-{beta} induced hyperglycosylation of prosaposin and its subsequent secretion, which ultimately caused depletion of lysosomal saposins. In melanoma patients, we found similar prosaposin hyperglycosylation in tumor-associated DCs, and reconstitution with prosaposin rescued activation of tumor-infiltrating T cells. Targeting tumor DCs with recombinant prosaposin triggered cancer protection and enhanced immune checkpoint therapy. Our studies demonstrate a critical function of prosaposin in tumor immunity and escape and introduce a novel principle of prosaposin-based cancer immunotherapy.

One Sentence SummaryProsaposin facilitates antigen cross-presentation and tumor immunity and its hyperglycosylation leads to immune evasion.
]]></description>
<dc:creator>Sharma, P.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Ly, K.</dc:creator>
<dc:creator>Kim, J. H.</dc:creator>
<dc:creator>Wan, Q.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Lou, M.</dc:creator>
<dc:creator>Kain, L.</dc:creator>
<dc:creator>Teyton, L.</dc:creator>
<dc:creator>Winau, F.</dc:creator>
<dc:date>2023-06-14</dc:date>
<dc:identifier>doi:10.1101/2023.06.14.545005</dc:identifier>
<dc:title><![CDATA[Hyperglycosylation of prosaposin in tumor DCs promotes immune escape in cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.14.544860v1?rss=1">
<title>
<![CDATA[
Participant-derived cell line transcriptomic analyses and mouse studies reveal a role for ZNF335 in plasma cholesterol statin response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.14.544860v1?rss=1</link>
<description><![CDATA[
BackgroundStatins lower circulating low-density lipoprotein cholesterol (LDLC) levels and reduce cardiovascular disease risk. Though highly efficacious in general, there is considerable inter-individual variation in statin efficacy that remains largely unexplained.

MethodsTo identify novel genes that may modulate statin-induced LDLC lowering, we used RNA-sequencing data from 426 control- and 2 M simvastatin-treated lymphoblastoid cell lines (LCLs) derived from European and African American ancestry participants of the Cholesterol and Pharmacogenetics (CAP) 40 mg/day 6-week simvastatin clinical trial (ClinicalTrials.gov Identifier: NCT00451828). We correlated statin-induced changes in LCL gene expression with plasma LDLC statin response in the corresponding CAP participants. For the most correlated gene identified (ZNF335), we followed up in vivo by comparing plasma cholesterol levels, lipoprotein profiles, and lipid statin response between wild-type mice and carriers of a hypomorphic (partial loss of function) missense mutation in Zfp335 (the mouse homolog of ZNF335).

ResultsThe statin-induced expression changes of 147 human LCL genes were significantly correlated to the plasma LDLC statin responses of the corresponding CAP participants in vivo (FDR=5%). The two genes with the strongest correlations were zinc finger protein 335 (ZNF335 aka NIF-1, rho=0.237, FDR-adj p=0.0085) and CCR4-NOT transcription complex subunit 3 (CNOT3, rho=0.233, FDR-adj p=0.0085). Chow-fed mice carrying a hypomorphic missense (R1092W; aka bloto) mutation in Zfp335 had significantly lower non-HDL cholesterol levels than wild type C57BL/6J mice in a sex combined model (p=0.04). Furthermore, male (but not female) mice carrying the Zfp335R1092W allele had significantly lower total and HDL cholesterol levels than wild-type mice. In a separate experiment, wild-type mice fed a control diet for 4 weeks and a matched simvastatin diet for an additional 4 weeks had significant statin-induced reductions in non-HDLC (-43{+/-}18% and -23{+/-}19% for males and females, respectively). Wild-type male (but not female) mice experienced significant reductions in plasma LDL particle concentrations, while male mice carrying Zfp335R1092W allele(s) exhibited a significantly blunted LDL statin response.

ConclusionsOur in vitro and in vivo studies identified ZNF335 as a novel modulator of plasma cholesterol levels and statin response, suggesting that variation in ZNF335 activity could contribute to inter-individual differences in statin clinical efficacy.
]]></description>
<dc:creator>Theusch, E.</dc:creator>
<dc:creator>Ting, F. Y.</dc:creator>
<dc:creator>Qin, Y.</dc:creator>
<dc:creator>Stevens, K.</dc:creator>
<dc:creator>Naidoo, D.</dc:creator>
<dc:creator>King, S. M.</dc:creator>
<dc:creator>Yang, N.</dc:creator>
<dc:creator>Orr, J.</dc:creator>
<dc:creator>Han, B. Y.</dc:creator>
<dc:creator>Cyster, J. G.</dc:creator>
<dc:creator>Chen, Y.-D. I.</dc:creator>
<dc:creator>Rotter, J. I.</dc:creator>
<dc:creator>Krauss, R. M.</dc:creator>
<dc:creator>Medina, M. W.</dc:creator>
<dc:date>2023-06-15</dc:date>
<dc:identifier>doi:10.1101/2023.06.14.544860</dc:identifier>
<dc:title><![CDATA[Participant-derived cell line transcriptomic analyses and mouse studies reveal a role for ZNF335 in plasma cholesterol statin response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.16.545386v1?rss=1">
<title>
<![CDATA[
CRISPR screens in iPSC-derived neurons reveal principles of tau proteostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.16.545386v1?rss=1</link>
<description><![CDATA[
Aggregation of the protein tau is a hallmark of Alzheimers disease and other tauopathies. Specific neuronal subtypes are selectively vulnerable to tau aggregation, but the underlying mechanisms are unknown. To systematically uncover the cellular factors controlling the accumulation of tau aggregates in human neurons, we conducted a genome-wide CRISPRi-based modifier screen in iPSC-derived neurons. The screen uncovered expected pathways, including autophagy, but also unexpected pathways, including UFMylation and GPI anchor synthesis, that control tau oligomer levels. We discover that the E3 ubiquitin ligase CUL5SOCS4 is a potent modifier of tau levels in human neurons, ubiquitinates tau, and is correlated with resilience to tauopathies in human disease. Disruption of mitochondrial function promotes proteasomal misprocessing of tau, which generates tau proteolytic fragments like those in disease and changes tau aggregation in vitro. These results reveal new principles of tau proteostasis in human neurons and pinpoint potential therapeutic targets for tauopathies.
]]></description>
<dc:creator>Samelson, A. J.</dc:creator>
<dc:creator>Ariqat, N.</dc:creator>
<dc:creator>McKetney, J.</dc:creator>
<dc:creator>Rohanitazangi, G.</dc:creator>
<dc:creator>Parra Bravo, C.</dc:creator>
<dc:creator>Goodness, D.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Grosjean, P.</dc:creator>
<dc:creator>Abskharon, R.</dc:creator>
<dc:creator>Eisenberg, D.</dc:creator>
<dc:creator>Kanaan, N. M.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:date>2023-06-17</dc:date>
<dc:identifier>doi:10.1101/2023.06.16.545386</dc:identifier>
<dc:title><![CDATA[CRISPR screens in iPSC-derived neurons reveal principles of tau proteostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.17.545426v1?rss=1">
<title>
<![CDATA[
Non-coding sequence variation reveals fragility within interleukin 2 feedback circuitry and shapes autoimmune disease risk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.17.545426v1?rss=1</link>
<description><![CDATA[
Genetic variants associated with human autoimmune diseases commonly map to non-coding control regions, particularly enhancers that function selectively in immune cells and fine-tune gene expression within a relatively narrow range of values. How such modest, cell-type-selective changes can meaningfully shape organismal disease risk remains unclear. To explore this issue, we experimentally manipulated species-conserved enhancers within the disease-associated IL2RA locus and studied accompanying changes in the progression of autoimmunity. Perturbing distinct enhancers with restricted activity in conventional T cells (Tconvs) or regulatory T cells (Tregs)--two functionally antagonistic T cell subsets--caused only modest, cell-type-selective decreases in IL2ra expression parameters. However, these same perturbations had striking and opposing effects in vivo, completely preventing or severely accelerating disease in a murine model of type 1 diabetes. Quantitative tissue imaging and computational modelling revealed that each enhancer manipulation impinged on distinct IL-2-dependent feedback circuits. These imbalances altered the intracellular signaling and intercellular communication dynamics of activated Tregs and Tconvs, producing opposing spatial domains that amplified or constrained ongoing autoimmune responses. These findings demonstrate how subtle changes in gene regulation stemming from non-coding variation can propagate across biological scales due to non-linearities in intra- and intercellular feedback circuitry, dramatically shaping disease risk at the organismal level.
]]></description>
<dc:creator>Simeonov, D. R.</dc:creator>
<dc:creator>Park, K.</dc:creator>
<dc:creator>Cortez, J. T.</dc:creator>
<dc:creator>Young, A.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Nguyen, V.</dc:creator>
<dc:creator>Umhoefer, J.</dc:creator>
<dc:creator>Indart, A. C.</dc:creator>
<dc:creator>Woo, J.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>Tsang, J. S.</dc:creator>
<dc:creator>Germain, R. N.</dc:creator>
<dc:creator>Wong, H. S.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2023-06-18</dc:date>
<dc:identifier>doi:10.1101/2023.06.17.545426</dc:identifier>
<dc:title><![CDATA[Non-coding sequence variation reveals fragility within interleukin 2 feedback circuitry and shapes autoimmune disease risk]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.19.545641v1?rss=1">
<title>
<![CDATA[
Stimulus-specific enhancement of responses in mouse primary visual cortex mediated by GABA release from VIP cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.19.545641v1?rss=1</link>
<description><![CDATA[
When adult mice are repeatedly exposed to a particular visual stimulus for as little as one hour per day for several days while their visual cortex (V1) is in the high-gain state produced by locomotion, that specific stimulus elicits much stronger responses in V1 neurons for the following several weeks, even when measured in anesthetized animals. Such stimulus-specific enhancement (SSE) is not seen if locomotion is prevented. The effect of locomotion on cortical responses is mediated by vasoactive intestinal peptide (VIP) positive interneurons, which can release both the peptide and the inhibitory neurotransmitter GABA. Here we used genetic ablation to determine which of those molecules secreted by VIP-ergic neurons is responsible for SSE. SSE was not impaired by VIP deletion but was prevented by compromising release of GABA from VIP cells. This finding suggests that SSE may result from Hebbian mechanisms that remain present in adult V1.

SIGNIFICANCEMany neurons package and release a peptide along with a conventional neurotransmitter. The conventional view is that such peptides exert late, slow effects on plasticity. We studied a form of cortical plasticity that depends on the activity of neurons that express both vasoactive intestinal peptide (VIP) and the inhibitory neurotransmitter GABA. GABA release accounted for their action on plasticity, with no effect of deleting the peptide on this phenomenon.
]]></description>
<dc:creator>Kaneko, M.</dc:creator>
<dc:creator>Hoseini, M. S.</dc:creator>
<dc:creator>Waschek, J. A.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:date>2023-06-20</dc:date>
<dc:identifier>doi:10.1101/2023.06.19.545641</dc:identifier>
<dc:title><![CDATA[Stimulus-specific enhancement of responses in mouse primary visual cortex mediated by GABA release from VIP cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.20.545796v1?rss=1">
<title>
<![CDATA[
Functionalized graphene-oxide grids enable high-resolution cryo-EM structures of the SNF2h-nucleosome complex without crosslinking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.20.545796v1?rss=1</link>
<description><![CDATA[
Single-particle cryo-EM is widely used to determine enzyme-nucleosome complex structures. However, cryo-EM sample preparation remains challenging and inconsistent due to complex denaturation at the air-water interface (AWI). To address this issue, we developed graphene-oxide-coated EM grids functionalized with either single-stranded DNA (ssDNA) or thiol-poly(acrylic acid-co-styrene) (TAASTY) co-polymer. These grids protect complexes between the chromatin remodeler SNF2h and nucleosomes from the AWI and facilitated collection of high-quality micrographs of intact SNF2h-nucleosome complexes in the absence of crosslinking. The data yields maps ranging from 2.3 to 3 [A] in resolution. 3D variability analysis reveals nucleotide-state linked conformational changes in SNF2h bound to a nucleosome. In addition, the analysis provides structural evidence for asymmetric coordination between two SNF2h protomers acting on the same nucleosome. We envision these grids will enable similar detailed structural analyses for other enzyme-nucleosome complexes and possibly other protein-nucleic acid complexes in general.
]]></description>
<dc:creator>Chio, U. S.</dc:creator>
<dc:creator>Palovcak, E.</dc:creator>
<dc:creator>Autzen, A. A. A.</dc:creator>
<dc:creator>Autzen, H. E.</dc:creator>
<dc:creator>Munoz, E. N.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Agard, D. A.</dc:creator>
<dc:creator>Armache, J.-P.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:date>2023-06-20</dc:date>
<dc:identifier>doi:10.1101/2023.06.20.545796</dc:identifier>
<dc:title><![CDATA[Functionalized graphene-oxide grids enable high-resolution cryo-EM structures of the SNF2h-nucleosome complex without crosslinking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.19.544278v1?rss=1">
<title>
<![CDATA[
Human iPSC 4R tauopathy model uncovers modifiers of tau propagation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.19.544278v1?rss=1</link>
<description><![CDATA[
Tauopathies are age-associated neurodegenerative diseases whose mechanistic underpinnings remain elusive, partially due to lack of appropriate human models. Current human induced pluripotent stem cell (hiPSC)-derived neurons express very low levels of 4-repeat (4R)-tau isoforms that are normally expressed in adult brain. Here, we engineered new iPSC lines to express 4R-tau and 4R-tau carrying the P301S MAPT mutation when differentiated into neurons. 4R-P301S neurons display progressive Tau inclusions upon seeding with Tau fibrils and recapitulate features of tauopathy phenotypes, including shared transcriptomic signatures, autophagic body accumulation, and impaired neuronal activity. A CRISPRi screen of genes associated with Tau pathobiology identified over 500 genetic modifiers of Tau-seeding-induced Tau propagation, including retromer VPS29 and the UFMylation cascade as top modifiers. In AD brains, the UFMylation cascade is altered in neurofibrillary-tangle-bearing neurons. Inhibiting the UFMylation cascade suppressed seeding-induced Tau propagation. This model provides a powerful platform to identify novel therapeutic strategies for 4R tauopathy.
]]></description>
<dc:creator>Parra Bravo, C.</dc:creator>
<dc:creator>Giani, A. M.</dc:creator>
<dc:creator>Madero-Perez, J.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Samelson, A. J.</dc:creator>
<dc:creator>Wong, M. Y.</dc:creator>
<dc:creator>Evangelisti, A.</dc:creator>
<dc:creator>Fan, L.</dc:creator>
<dc:creator>Pozner, T.</dc:creator>
<dc:creator>Mercedes, M.</dc:creator>
<dc:creator>Ye, P.</dc:creator>
<dc:creator>Patel, T.</dc:creator>
<dc:creator>Yarahmady, A.</dc:creator>
<dc:creator>Carling, G.</dc:creator>
<dc:creator>Lee, V. M. Y.</dc:creator>
<dc:creator>Sharma, M.</dc:creator>
<dc:creator>Mok, S.-A.</dc:creator>
<dc:creator>Luo, W.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Gong, S.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:date>2023-06-22</dc:date>
<dc:identifier>doi:10.1101/2023.06.19.544278</dc:identifier>
<dc:title><![CDATA[Human iPSC 4R tauopathy model uncovers modifiers of tau propagation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.19.545618v1?rss=1">
<title>
<![CDATA[
GATA-regulated transcriptional program dictate cell fate equilibrium to establish the maternal-fetal exchange interface and fetal development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.19.545618v1?rss=1</link>
<description><![CDATA[
The placenta establishes a maternal-fetal exchange interface to transport nutrients and gases between the mother and the fetus. Establishment of this exchange interface relies on the development of multinucleated syncytiotrophoblasts (SynT) from trophoblast progenitors and defect in SynT development often leads to pregnancy failure and impaired embryonic development. Here, we show that mouse embryos with conditional deletion of GATA2 and GATA3 in labyrinth trophoblast progenitors have underdeveloped placenta and die [~] embryonic day 9.5 (E9.5). Single cell RNA Seq (scRNA-Seq) analysis revealed excessive accumulation of multipotent labyrinth trophoblast progenitors upon conditional deletion of GATA factors. The GATA factor-deleted multipotent progenitors were unable to differentiate to matured SynTs. We also show that the GATA factor-mediated priming of trophoblast progenitors for SynT differentiation is a conserved event during human placentation. Loss of either GATA2 or GATA3 in cytotrophoblast (CTB)-derived human trophoblast stem cells (human TSCs) drastically inhibits SynT differentiation potential. Identification of GATA2 and GATA3 target genes along with comparative bioinformatics analyses revealed that GATA factors directly regulate hundreds of common genes in human TSCs, including genes that are essential for SynT development and implicated in preeclampsia and fetal growth retardation. Thus, our study uncovers a conserved molecular mechanism, in which coordinated function of GATA2 and GATA3 promote trophoblast progenitor-to-SynT commitment ensuring establishment of the maternal-fetal exchange interface.
]]></description>
<dc:creator>Ghosh, A.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Kumar, R. P.</dc:creator>
<dc:creator>Ray, S.</dc:creator>
<dc:creator>Saha, A.</dc:creator>
<dc:creator>Roy, N.</dc:creator>
<dc:creator>Dasgupta, P.</dc:creator>
<dc:creator>Marsh, C.</dc:creator>
<dc:creator>Paul, S.</dc:creator>
<dc:date>2023-06-22</dc:date>
<dc:identifier>doi:10.1101/2023.06.19.545618</dc:identifier>
<dc:title><![CDATA[GATA-regulated transcriptional program dictate cell fate equilibrium to establish the maternal-fetal exchange interface and fetal development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.21.546024v1?rss=1">
<title>
<![CDATA[
Small-field visual projection neurons detect translational optic flow and support walking control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.21.546024v1?rss=1</link>
<description><![CDATA[
Animals rely on visual motion for navigating the world, and research in flies has clarified how neural circuits extract information from moving visual scenes. However, the major pathways connecting these patterns of optic flow to behavior remain poorly understood. Using a high-throughput quantitative assay of visually guided behaviors and genetic neuronal silencing, we discovered a region in Drosophilas protocerebrum critical for visual motion following. We used neuronal silencing, calcium imaging, and optogenetics to identify a single cell type, LPC1, that innervates this region, detects translational optic flow, and plays a key role in regulating forward walking. Moreover, the population of LPC1s can estimate the travelling direction, such as when gaze direction diverges from body heading. By linking specific cell types and their visual computations to specific behaviors, our findings establish a foundation for understanding how the nervous system uses vision to guide navigation.
]]></description>
<dc:creator>Isaacson, M. D.</dc:creator>
<dc:creator>Eliason, J. L.</dc:creator>
<dc:creator>Nern, A.</dc:creator>
<dc:creator>Rogers, E. M.</dc:creator>
<dc:creator>Lott, G. K.</dc:creator>
<dc:creator>Tabachnik, T.</dc:creator>
<dc:creator>Rowell, W. J.</dc:creator>
<dc:creator>Edwards, A. W.</dc:creator>
<dc:creator>Korff, W. L.</dc:creator>
<dc:creator>Rubin, G. M.</dc:creator>
<dc:creator>Branson, K.</dc:creator>
<dc:creator>Reiser, M. B.</dc:creator>
<dc:date>2023-06-22</dc:date>
<dc:identifier>doi:10.1101/2023.06.21.546024</dc:identifier>
<dc:title><![CDATA[Small-field visual projection neurons detect translational optic flow and support walking control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.21.543782v1?rss=1">
<title>
<![CDATA[
Tiny Earth CURE improves student persistence in science 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.21.543782v1?rss=1</link>
<description><![CDATA[
Course-based undergraduate research experiences (CUREs) enhance student retention in STEM, particularly among students who belong to historically excluded communities. Yet the mechanisms by which CUREs contribute to student integration and persistence are poorly understood. Utilizing the Tripartite Integration Model of Social Influence (TIMSI), this longitudinal study examines how Tiny Earth, an antibiotic-discovery CURE, impacts students scientific self-efficacy, scientific identity, endorsement of scientific community values, and intentions to persist in science. The study also explores how gains in TIMSI factors vary as a function of student demographics and course characteristics. Results of pre-and post-course measurements show that scientific self-efficacy and identity increased among students in Tiny Earth, and some student demographics and course characteristics moderated these gains. Gains in scientific self-efficacy, identity, and values correlated with gains in persistence intentions, whereas student demographics and course characteristics did not. Results of this study show that the Tiny Earth CURE enhanced students integration into the scientific community, which was linked to intentions of students of both historically underrepresented and majority groups to persist in STEM. We discuss how courses that provide opportunity to learn science skills in the context of a CURE can contribute toward enlarging and diversifying the STEM workforce.
]]></description>
<dc:creator>Miller, S.</dc:creator>
<dc:creator>Aldana, C. C.</dc:creator>
<dc:creator>Du, W.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Maldonado, N.</dc:creator>
<dc:creator>Sandoval, P.</dc:creator>
<dc:creator>Vong, J.</dc:creator>
<dc:creator>Young, G.</dc:creator>
<dc:creator>Handelsman, J.</dc:creator>
<dc:creator>Broderick, N. A.</dc:creator>
<dc:creator>Hernandez, P. R.</dc:creator>
<dc:creator>Estrada, M.</dc:creator>
<dc:date>2023-06-23</dc:date>
<dc:identifier>doi:10.1101/2023.06.21.543782</dc:identifier>
<dc:title><![CDATA[Tiny Earth CURE improves student persistence in science]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.23.546285v1?rss=1">
<title>
<![CDATA[
Activation of Shh/Smo is sufficient to maintain oligodendrocyte precursor cells in an undifferentiated state but is not necessary for differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.23.546285v1?rss=1</link>
<description><![CDATA[
Myelination is the terminal step in a complex and precisely timed program that orchestrates the proliferation, migration and differentiation of oligodendroglial cells. It is thought that Sonic Hedgehog (Shh) acting on Smoothened (Smo) participates in regulating this process, but that these effects are highly context dependent. Here, we investigate oligodendroglial development and remyelination from three specific transgenic lines: NG2-CreERT2 (control), Smofl/fl/NG2-CreERT2 (loss of function) and SmoM2/NG2-CreERT2 (gain of function), as well as pharmacological manipulation that enhance or inhibit the Smo pathway (SAG or cyclopamine treatment respectively). To explore the effects of Shh/Smo on differentiation and myelination in vivo, we developed a highly quantifiable model by transplanting OPCs in the retina. We find that myelination is greatly enhanced upon cyclopamine treatment and hypothesize that Shh/Smo could promote OPC proliferation to subsequently inhibit differentiation. Consistent with this hypothesis, we find that the genetic activation of Smo significantly increased numbers of OPCs and decreased oligodendrocyte differentiation when we examined the corpus callosum during development and after cuprizone demyelination and remyelination. However, upon loss of function with the conditional ablation of Smo, myelination in the same scenarios are unchanged. Taken together, our present findings suggest that the Shh pathway is sufficient to maintain OPCs in an undifferentiated state, but is not necessary for myelination and remyelination.
]]></description>
<dc:creator>Nocera, S.</dc:creator>
<dc:creator>Marchena, M. A.</dc:creator>
<dc:creator>Fernandez-Gomez, B.</dc:creator>
<dc:creator>Lao, Y.</dc:creator>
<dc:creator>Cordano, C.</dc:creator>
<dc:creator>Gomez-Torres, O.</dc:creator>
<dc:creator>Lujan, R.</dc:creator>
<dc:creator>de Castro, F.</dc:creator>
<dc:date>2023-06-23</dc:date>
<dc:identifier>doi:10.1101/2023.06.23.546285</dc:identifier>
<dc:title><![CDATA[Activation of Shh/Smo is sufficient to maintain oligodendrocyte precursor cells in an undifferentiated state but is not necessary for differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.23.546186v1?rss=1">
<title>
<![CDATA[
The unique microbial diversity, community composition and networks of Pacific Islander endocervical and vaginal microbiomes in the presence or absence of Chlamydia trachomatis infection using metagenomic shotgun sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.23.546186v1?rss=1</link>
<description><![CDATA[
BackgroundPacific Islanders are a vulnerable population with a high prevalence of Chlamydia trachomatis (Ct) sexually transmitted infections (STIs) and remain underrepresented in research. Here, 258 vaginal and 92 paired endocervical samples from women of diverse ethnicities in Fiji were evaluated using metagenomics to characterize microbial relative abundance, composition and networks including associations with Neisseria gonorrhoeae, human papilloma virus (HPV), Mycoplasma genitalium, Candida and bacterial vaginosis (BV).

ResultsPacific Islander ethnicities and age <25 years were significantly associated with Ct infection. Using VALENCIA, a sub-community state type (subCST) classifier, 93 (36.04%) vaginal and 38 (41.30%) endocervical microbiomes did not match reference subCSTs. Four unique subCSTs were developed to better classify Pacific Islander microbiomes: IV-D0, dominated by Gardnerella vaginalis; IV-D1, high/moderate G. vaginalis relative abundance with Prevotella spp.; IV-D2, high/moderate G. vaginalis with Lactobacillus iners; and IV-E, moderate Prevotella spp. Ct positive endocervical and paired vaginal microbiomes were significantly more likely to have differential species relative abundance (81.58%) than Ct negative pairs (35.84%; AOR: 7.93; 95% CI: 2.93-21.93; P<0.0001). A significantly higher alpha diversity was found for iTaukei ethnicity, all subCST-IV types, BV, and Ct for vaginal microbiomes. For endocervical microbiomes, higher diversity was significant for subCST-IV-A, subCST-IV-D1, and subCST-IV-E, and high-risk HPV types. Overall, there was a significantly higher diversity for the endocervix in paired microbiomes. Vaginal microbiomes showed significant divergence in community composition as above and for Candida. Endocervical composition varied significantly by subCST type and Ct status. Gut and BV-associated bacterial clusters were present in Ct positive and negative paired endocervical and vaginal microbiome networks but were smaller with fewer bacterial and no Lactobacillus spp. interactions in Ct-infected endocervical networks where G. vaginalis generated polymicrobial biofilms along with Ct likely influence pathogenicity.

ConclusionsFijian endocervical and vaginal microbiomes represent divergent microbial abundance and compositions, especially for Pacific Islander ethnicities, with distinct subCSTs compared to other global populations. The higher microbial diversity of the endocervix with prevalent G. vaginalis, L. iners and Prevotella spp. suggest that these microbiomes/networks may predispose to and/or promote chlamydial and HPV pathogenesis. Prospective studies are needed to further define causal associations to develop successful interventions.
]]></description>
<dc:creator>Bommana, S.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Kama, M.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Kodimerla, R.</dc:creator>
<dc:creator>Jijakli, K.</dc:creator>
<dc:creator>Read, T.</dc:creator>
<dc:creator>Dean, D.</dc:creator>
<dc:date>2023-06-24</dc:date>
<dc:identifier>doi:10.1101/2023.06.23.546186</dc:identifier>
<dc:title><![CDATA[The unique microbial diversity, community composition and networks of Pacific Islander endocervical and vaginal microbiomes in the presence or absence of Chlamydia trachomatis infection using metagenomic shotgun sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.24.545427v1?rss=1">
<title>
<![CDATA[
Diverse viral cas genes antagonize CRISPR immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.24.545427v1?rss=1</link>
<description><![CDATA[
Prokaryotic CRISPR-Cas immunity is subverted via anti-CRISPRs (Acrs), small proteins that inhibit Cas protein activities when expressed during the phage lytic cycle or from resident prophages or plasmids. CRISPR-Cas defenses are classified into 6 types and 33 subtypes, which employ a diverse suite of Cas effectors and differ in their mechanisms of interference. As Acrs often work via binding to a cognate Cas protein, inhibition is almost always limited to a single CRISPR type. Furthermore, while acr genes are frequently organized together in phage-associated gene clusters, how such inhibitors initially evolve has remained unclear. Here we have investigated the Acr content and inhibition specificity of a collection of Listeria isolates, which naturally harbor four diverse CRISPR-Cas systems (types I-B, II-A, II-C, and VI-A). We observed widespread antagonism of CRISPR, which we traced to 12 novel and 4 known Acr gene families encoded on endogenous mobile genetic elements. Among these were two Acrs that possess sequence homology to type I-B Cas proteins and assemble into a defective interference complex. Surprisingly, an additional type I-B Cas homolog did not affect type I immunity, but instead inhibited the RNA-targeting type VI CRISPR system through sequestration of crRNA. By probing the IMGVR database of viral genomes, we detected abundant orphan cas genes located within putative anti-defense gene clusters. We experimentally verified the Acr activity of one viral cas gene, a particularly broad-spectrum cas3 homolog that inhibits type I-B, II-A, and VI-A CRISPR immunity. Our observations provide direct evidence of Acr evolution via cas gene co-option, and new genes with potential for broad-spectrum control of genome editing technologies.
]]></description>
<dc:creator>Katz, M. A.</dc:creator>
<dc:creator>Sawyer, E. M.</dc:creator>
<dc:creator>Kozlova, A.</dc:creator>
<dc:creator>Williams, M. C.</dc:creator>
<dc:creator>Margolis, S. R.</dc:creator>
<dc:creator>Oriolt, L.</dc:creator>
<dc:creator>Johnson, M.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Meeske, A. J.</dc:creator>
<dc:date>2023-06-24</dc:date>
<dc:identifier>doi:10.1101/2023.06.24.545427</dc:identifier>
<dc:title><![CDATA[Diverse viral cas genes antagonize CRISPR immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.22.546152v1?rss=1">
<title>
<![CDATA[
Enabling Success Through Transformative Practices in STEM: The Effects of Applying an Anti-Deficit Framework to Diversity and Equity Programming 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.22.546152v1?rss=1</link>
<description><![CDATA[
Diversity, equity, and inclusion (DEI) programming and literature for historically underrepresented and marginalized students in science, technology, engineering, and mathematics degree programs often focuses on illuminating the challenges they face. The repeated emphasis on negative experiences creates a deficit-focused thread of inquiry that may unintentionally reinscribe persistent disparities and inequities. In this practice brief, we describe the positive effect of adapting anti-deficit framework with social career cognitive theory in developing and evaluating an Initiative for Maximizing Student Development program at a biomedical sciences graduate school, as well as how other institutions can explore, implement, and evaluate transformative DEI practices. In identifying and emphasizing the enablers of success while facilitating structured opportunities for personal and professional identity development, students and program leadership align values and goals to increase academic and scientific development, as well as community and social support.
]]></description>
<dc:creator>Duncan, D. S.</dc:creator>
<dc:creator>Jackson, J. L.</dc:creator>
<dc:creator>Collins, S.</dc:creator>
<dc:creator>Teherani, A.</dc:creator>
<dc:date>2023-06-25</dc:date>
<dc:identifier>doi:10.1101/2023.06.22.546152</dc:identifier>
<dc:title><![CDATA[Enabling Success Through Transformative Practices in STEM: The Effects of Applying an Anti-Deficit Framework to Diversity and Equity Programming]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.23.546317v1?rss=1">
<title>
<![CDATA[
The microenvironment dictates glycocalyx construction and immune surveillance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.23.546317v1?rss=1</link>
<description><![CDATA[
Efforts to identify anti-cancer therapeutics and understand tumor-immune interactions are built with in vitro models that do not match the microenvironmental characteristics of human tissues. Using in vitro models which mimic the physical properties of healthy or cancerous tissues and a physiologically relevant culture medium, we demonstrate that the chemical and physical properties of the microenvironment regulate the composition and topology of the glycocalyx. Remarkably, we find that cancer and age-related changes in the physical properties of the microenvironment are sufficient to adjust immune surveillance via the topology of the glycocalyx, a previously unknown phenomenon observable only with a physiologically relevant culture medium.

Key PointsO_LICulture medium dictates cellular mechanoresponse signatures in vitro
C_LIO_LIEpithelial glycocalyx construction is mediated by Heat Shock Factor 1 (HSF1)
C_LIO_LISialic acid topology dictates Natural Killer cell cytotoxicity
C_LIO_LIPhysiological microenvironments reveal distinct glycobiology
C_LI
]]></description>
<dc:creator>Tharp, K.</dc:creator>
<dc:creator>Park, S.</dc:creator>
<dc:creator>Timblin, G. A.</dc:creator>
<dc:creator>Richards, A.</dc:creator>
<dc:creator>Berg, J.</dc:creator>
<dc:creator>Twells, N.</dc:creator>
<dc:creator>Riley, N. M.</dc:creator>
<dc:creator>Peltan, E.</dc:creator>
<dc:creator>Shon, D. J.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Tsui, C. K.</dc:creator>
<dc:creator>Palomba, F.</dc:creator>
<dc:creator>Lefebvre, A. E.</dc:creator>
<dc:creator>Soens, R.</dc:creator>
<dc:creator>Ayad, N.</dc:creator>
<dc:creator>ten Hoeve-Scott, J.</dc:creator>
<dc:creator>Healy, K.</dc:creator>
<dc:creator>Digman, M.</dc:creator>
<dc:creator>Dillin, A.</dc:creator>
<dc:creator>Bertozzi, C.</dc:creator>
<dc:creator>Mahal, L. K.</dc:creator>
<dc:creator>Swaney, D.</dc:creator>
<dc:creator>Cantor, J. R.</dc:creator>
<dc:creator>Paszek, M.</dc:creator>
<dc:creator>Weaver, V.</dc:creator>
<dc:date>2023-06-25</dc:date>
<dc:identifier>doi:10.1101/2023.06.23.546317</dc:identifier>
<dc:title><![CDATA[The microenvironment dictates glycocalyx construction and immune surveillance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.25.546454v1?rss=1">
<title>
<![CDATA[
Prolonged Kaposi's Sarcoma-associated Herpesvirus vIL-6 Exposure Enhances Inflammatory Responses by Epigenetic Reprogramming 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.25.546454v1?rss=1</link>
<description><![CDATA[
Kaposi sarcoma-associated herpesvirus (KSHV) inflammatory cytokine syndrome (KICS) is a newly described chronic inflammatory disease condition caused by KSHV infection and is characterized by high KSHV viral load and sustained elevations of serum KSHV-encoded IL-6 (vIL-6) and human IL-6 (hIL-6). KICS has significant immortality and possesses greater risks of having other complications, which include malignancies. Although prolonged inflammatory vIL-6 exposure by persistent KSHV infection is expected to have key roles in subsequent disease development, the biological effects of prolonged vIL-6 exposure remain elusive. Using thiol-Linked Alkylation for the Metabolic Sequencing and Cleavage Under Target & Release Using Nuclease analysis, we studied the effect of prolonged vIL-6 exposure in chromatin landscape and resulting cytokine production. The studies showed that prolonged vIL-6 exposure increased Bromodomain containing 4 (BRD4) and histone H3 lysine 27 acetylation co-occupancies on chromatin, and the recruitment sites were frequently co-localized with poised RNAPII with associated enzymes. Increased BRD4 recruitment on promoters was associated with increased and prolonged NF-KB p65 binding after the lipopolysaccharide stimulation. The p65 binding resulted in quicker and sustained transcription bursts from the promoters; this mechanism increased total amounts of hIL-6 and IL-10 in tissue culture. Pretreatment with the BRD4 inhibitor, OTX015, eliminated the enhanced inflammatory cytokine production. These findings suggest that persistent vIL-6 exposure may establish a chromatin landscape favorable for the reactivation of inflammatory responses in monocytes. This epigenetic memory may explain the greater risk of chronic inflammatory disease development in KSHV-infected individuals.

Author summaryCombined and continuous cytokine stimulation triggers transcription reprogramming and is often used for specific tissue development. Continuous vIL-6 exposure occurs in KSHV-infected patients and leads to inflammatory cytokine storm with high mortality. However, possible epigenetic reprogramming by the vIL-6 and its association with pathogenesis remain unclear. Here we demonstrate the establishment of a new chromatin landscape mediated by BRD4 through prolonged vIL-6 exposure which contributes to more robust and rapid transcription and increased cytokines production. Inhibition of BRD4 suppressed this inflammatory response. Our results indicate that targeting the epigenetic effect of viral cytokines may lead to novel therapies for KSHV-induced inflammatory cytokine storms.
]]></description>
<dc:creator>Inagaki, T.</dc:creator>
<dc:creator>Wang, K.-H.</dc:creator>
<dc:creator>Kumar, A.</dc:creator>
<dc:creator>Izumiya, C.</dc:creator>
<dc:creator>Miura, H.</dc:creator>
<dc:creator>Komaki, S.</dc:creator>
<dc:creator>Davis, R. R.</dc:creator>
<dc:creator>Tepper, C.</dc:creator>
<dc:creator>Katano, H.</dc:creator>
<dc:creator>Shimoda, M.</dc:creator>
<dc:creator>Izumiya, Y.</dc:creator>
<dc:date>2023-06-25</dc:date>
<dc:identifier>doi:10.1101/2023.06.25.546454</dc:identifier>
<dc:title><![CDATA[Prolonged Kaposi's Sarcoma-associated Herpesvirus vIL-6 Exposure Enhances Inflammatory Responses by Epigenetic Reprogramming]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.23.546302v1?rss=1">
<title>
<![CDATA[
Multi-night naturalistic cortico-basal recordings reveal mechanisms of NREM slow wave suppression and spontaneous awakenings in Parkinson's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.23.546302v1?rss=1</link>
<description><![CDATA[
BackgroundSleep disturbance is a prevalent and highly disabling comorbidity in individuals with Parkinsons disease (PD) that leads to worsening of daytime symptoms, accelerated disease progression and reduced quality of life.

ObjectivesWe aimed to investigate changes in sleep neurophysiology in PD particularly during non-rapid eye movement (NREM) sleep, both in the presence and absence of deep brain stimulation (DBS).

MethodsMulti-night (n=58) intracranial recordings were performed at-home, from chronic electrocorticography and subcortical electrodes, with sensing-enabled DBS pulse generators, paired with portable polysomnography. Four people with PD and one person with cervical dystonia were evaluated to determine the neural structures, signals and connections modulated during NREM sleep and prior to spontaneous awakenings. Recordings were performed both ON and OFF DBS in the presence of conventional dopaminergic replacement medications.

ResultsWe demonstrate an increase in cortico-basal slow wave activity in delta (1-4 Hz) and a decrease in beta (13-31 Hz) during NREM (N2 and N3) versus wakefulness in PD. Cortical-subcortical coherence was also found to be higher in the delta range and lower in the beta range during NREM versus wakefulness. DBS stimulation resulted in a further elevation in cortical delta and a decrease in alpha (8-13 Hz) and low beta (13-15 Hz) power compared to the OFF stimulation state. During NREM sleep, we observed a strong inverse interaction between subcortical beta and cortical slow wave activity and found that subcortical beta increases prior to spontaneous awakenings.

ConclusionsChronic, multi-night recordings in PD reveal opposing sleep stage specific modulations of cortico-basal slow wave activity in delta and subcortical beta power and connectivity in NREM, effects that are enhanced in the presence of DBS. Within NREM specifically, subcortical beta and cortical delta are strongly inversely correlated and subcortical beta power is found to increase prior to and predict spontaneous awakenings. We find that DBS therapy appears to improve sleep in PD partially through direct modulation of cortico-basal beta and delta oscillations. Our findings help elucidate a contributory mechanism responsible for sleep disturbances in PD and highlight potential biomarkers for future precision neuromodulation therapies targeting sleep and spontaneous awakenings.
]]></description>
<dc:creator>Anjum, M. F.</dc:creator>
<dc:creator>Smyth, C.</dc:creator>
<dc:creator>Dijk, D.-J.</dc:creator>
<dc:creator>Starr, P.</dc:creator>
<dc:creator>Denison, T.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.23.546302</dc:identifier>
<dc:title><![CDATA[Multi-night naturalistic cortico-basal recordings reveal mechanisms of NREM slow wave suppression and spontaneous awakenings in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.24.546411v1?rss=1">
<title>
<![CDATA[
TOMM40 and TOMM22 of the Translocase Outer Mitochondrial Membrane Complex rescue statin-impaired mitochondrial dynamics, morphology, and mitophagy in skeletal myotubes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.24.546411v1?rss=1</link>
<description><![CDATA[
BackgroundStatins are the drugs most commonly used for lowering plasma low-density lipoprotein (LDL) cholesterol levels and reducing cardiovascular disease risk. Although generally well tolerated, statins can induce myopathy, a major cause of non-adherence to treatment. Impaired mitochondrial function has been implicated as a cause of statin-induced myopathy, but the underlying mechanism remains unclear. We have shown that simvastatin downregulates transcription of TOMM40 and TOMM22, genes that encode major subunits of the translocase of outer mitochondrial membrane (TOM) complex which is responsible for importing nuclear-encoded proteins and maintaining mitochondrial function. We therefore investigated the role of TOMM40 and TOMM22 in mediating statin effects on mitochondrial function, dynamics, and mitophagy.

MethodsCellular and biochemical assays and transmission electron microscopy were used to investigate effects of simvastatin and TOMM40 and TOMM22 expression on measures of mitochondrial function and dynamics in C2C12 and primary human skeletal cell myotubes.

ResultsKnockdown of TOMM40 and TOMM22 in skeletal cell myotubes impaired mitochondrial oxidative function, increased production of mitochondrial superoxide, reduced mitochondrial cholesterol and CoQ levels, disrupted mitochondrial dynamics and morphology, and increased mitophagy, with similar effects resulting from simvastatin treatment. Overexpression of TOMM40 and TOMM22 in simvastatin-treated muscle cells rescued statin effects on mitochondrial dynamics, but not on mitochondrial function or cholesterol and CoQ levels. Moreover, overexpression of these genes resulted in an increase in number and density of cellular mitochondria.

ConclusionThese results confirm that TOMM40 and TOMM22 are central in regulating mitochondrial homeostasis and demonstrate that downregulation of these genes by statin treatment mediates disruption of mitochondrial dynamics, morphology, and mitophagy, effects that may contribute to statin-induced myopathy.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=102 SRC="FIGDIR/small/546411v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Yang, N. V.</dc:creator>
<dc:creator>Rogers, S.</dc:creator>
<dc:creator>Guerra, R.</dc:creator>
<dc:creator>Pagliarini, D. J.</dc:creator>
<dc:creator>Theusch, E.</dc:creator>
<dc:creator>Krauss, R. M.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.24.546411</dc:identifier>
<dc:title><![CDATA[TOMM40 and TOMM22 of the Translocase Outer Mitochondrial Membrane Complex rescue statin-impaired mitochondrial dynamics, morphology, and mitophagy in skeletal myotubes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.25.546328v1?rss=1">
<title>
<![CDATA[
Patterning and folding of intestinal villi by active mesenchymal dewetting 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.25.546328v1?rss=1</link>
<description><![CDATA[
Tissue folding generates structural motifs critical to organ function. In the intestine, bending of a flat epithelium into a periodic pattern of folds gives rise to villi, the numerous finger-like protrusions that are essential for nutrient absorption. However, the molecular and mechanical mechanisms driving the initiation and morphogenesis of villi remain a matter of debate. Here, we identify an active mechanical mechanism that simultaneously patterns and folds intestinal villi. We find that PDGFRA+ subepithelial mesenchymal cells generate myosin II-dependent forces sufficient to produce patterned curvature in neighboring tissue interfaces. At the cell-level, this occurs through a process dependent upon matrix metalloproteinase-mediated tissue fluidization and altered cell-ECM adhesion. By combining computational models with in vivo experiments, we reveal these cellular features manifest at the tissue-level as differences in interfacial tensions that promote mesenchymal aggregation and interface bending through a process analogous to the active de-wetting of a thin liquid film.
]]></description>
<dc:creator>Huycke, T. R.</dc:creator>
<dc:creator>Miyazaki, H.</dc:creator>
<dc:creator>Häkkinen, T.</dc:creator>
<dc:creator>Srivastava, V.</dc:creator>
<dc:creator>Barruet, E.</dc:creator>
<dc:creator>McGinnis, C. S.</dc:creator>
<dc:creator>Kalantari, A.</dc:creator>
<dc:creator>Cornwall-Scoones, J.</dc:creator>
<dc:creator>Vaka, D.</dc:creator>
<dc:creator>Zhu, Q.</dc:creator>
<dc:creator>Jo, H.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:creator>Garikipati, K.</dc:creator>
<dc:creator>Boffelli, D.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.25.546328</dc:identifier>
<dc:title><![CDATA[Patterning and folding of intestinal villi by active mesenchymal dewetting]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.26.546582v1?rss=1">
<title>
<![CDATA[
Cross regional coordination of neural activity in the human brain during autobiographical self-referential processing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.26.546582v1?rss=1</link>
<description><![CDATA[
For the human mind to operate, populations of neurons across remote regions of the brain need to coordinate their activity in the subsecond temporal scale. To date, our knowledge of such fast interactions involving cortical and subcortical structures in large brains, such as the human brain, remains limited. Here, we used stereo-electroencephalography (sEEG) recordings across four brain regions that are known, from decades of work, to be important for autobiographical memory processing. Our recordings involved 31 human participants implanted with intracranial electrodes in the hippocampus (HPC), posteromedial cortex (PMC), and ventromedial, as well as orbital subregions of the prefrontal cortex (OFC). In 14 subjects, we also recorded simultaneously in the anterior thalamus (ANT) across various experimental conditions and with direct electrical stimulations. Our observations provide new lines of correlative and causal evidence about the spatiotemporal profile of oscillatory coordination of cortical and subcortical activity during self-referential memory-based processing.
]]></description>
<dc:creator>Stieger, J.</dc:creator>
<dc:creator>Pinheiro-Chagas, P.</dc:creator>
<dc:creator>Fang, Y.</dc:creator>
<dc:creator>Lusk, Z.</dc:creator>
<dc:creator>Perry, C.</dc:creator>
<dc:creator>Wagner, A. D.</dc:creator>
<dc:creator>Contreras, D.</dc:creator>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>Huguenard, J.</dc:creator>
<dc:creator>Buch, V.</dc:creator>
<dc:creator>Parvizi, J.</dc:creator>
<dc:date>2023-06-27</dc:date>
<dc:identifier>doi:10.1101/2023.06.26.546582</dc:identifier>
<dc:title><![CDATA[Cross regional coordination of neural activity in the human brain during autobiographical self-referential processing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.27.545450v1?rss=1">
<title>
<![CDATA[
Population encoding of stimulus features along the visual hierarchy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.27.545450v1?rss=1</link>
<description><![CDATA[
The retina and primary visual cortex (V1) both exhibit diverse neural populations sensitive to diverse visual features. Yet it remains unclear how neural populations in each area partition stimulus space to span these features. One possibility is that neural populations are organized into discrete groups of neurons, with each group signaling a particular constellation of features. Alternatively, neurons could be continuously distributed across feature-encoding space. To distinguish these possibilities, we presented a battery of visual stimuli to mouse retina and V1 while measuring neural responses with multi-electrode arrays. Using machine learning approaches, we developed a manifold embedding technique that captures how neural populations partition feature space and how visual responses correlate with physiological and anatomical properties of individual neurons. We show that retinal populations discretely encode features, while V1 populations provide a more continuous representation. Applying the same analysis approach to convolutional neural networks that model visual processing, we demonstrate that they partition features much more similarly to the retina, indicating they are more like big retinas than little brains.
]]></description>
<dc:creator>Dyballa, L.</dc:creator>
<dc:creator>Rudzite, A. M.</dc:creator>
<dc:creator>Hoseini, M. S.</dc:creator>
<dc:creator>Thapa, M.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:creator>Field, G. D.</dc:creator>
<dc:creator>Zucker, S. W.</dc:creator>
<dc:date>2023-06-27</dc:date>
<dc:identifier>doi:10.1101/2023.06.27.545450</dc:identifier>
<dc:title><![CDATA[Population encoding of stimulus features along the visual hierarchy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.27.546744v1?rss=1">
<title>
<![CDATA[
Self-extinguishing relay waves enable homeostaticcontrol of human neutrophil swarming 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.27.546744v1?rss=1</link>
<description><![CDATA[
Neutrophils exhibit self-amplified swarming to sites of injury and infection. How swarming is controlled to ensure the proper level of neutrophil recruitment is unknown. Using an ex vivo model of infection, we find that human neutrophils use active relay to generate multiple pulsatile waves of swarming signals. Unlike classic active relay systems such as action potentials, neutrophil swarming relay waves are self-extinguishing, limiting the spatial range of cell recruitment. We identify an NADPH-oxidase-based negative feedback loop that is needed for this self-extinguishing behavior. Through this circuit, neutrophils adjust the number and size of swarming waves for homeostatic levels of cell recruitment over a wide range of initial cell densities. We link a broken homeostat to neutrophil over-recruitment in the context of human chronic granulomatous disease.
]]></description>
<dc:creator>Strickland, J.</dc:creator>
<dc:creator>Pan, D.</dc:creator>
<dc:creator>Godfrey, C.</dc:creator>
<dc:creator>Kim, J. S.</dc:creator>
<dc:creator>Hopke, A.</dc:creator>
<dc:creator>Degrange, M.</dc:creator>
<dc:creator>Villavicencio, B.</dc:creator>
<dc:creator>Mansour, M. K.</dc:creator>
<dc:creator>Zerbe, C. S.</dc:creator>
<dc:creator>Irimia, D.</dc:creator>
<dc:creator>Amir, A.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:date>2023-06-28</dc:date>
<dc:identifier>doi:10.1101/2023.06.27.546744</dc:identifier>
<dc:title><![CDATA[Self-extinguishing relay waves enable homeostaticcontrol of human neutrophil swarming]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.28.546958v1?rss=1">
<title>
<![CDATA[
Recruitment of BAG2 to DNAJ-PKAc scaffolds promotes cell survival and resistance to drug-induced apoptosis in fibrolamellar carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.28.546958v1?rss=1</link>
<description><![CDATA[
The DNAJ-PKAc fusion kinase is a defining feature of the adolescent liver cancer fibrolamellar carcinoma (FLC). A single lesion on chromosome 19 generates this mutant kinase by creating a fused gene encoding the chaperonin binding domain of Hsp40 (DNAJ) in frame with the catalytic core of protein kinase A (PKAc). FLC tumors are notoriously resistant to standard chemotherapies. Aberrant kinase activity is assumed to be a contributing factor. Yet recruitment of binding partners, such as the chaperone Hsp70, implies that the scaffolding function of DNAJ- PKAc may also underlie pathogenesis. By combining proximity proteomics with biochemical analyses and photoactivation live-cell imaging we demonstrate that DNAJ-PKAc is not constrained by A-kinase anchoring proteins. Consequently, the fusion kinase phosphorylates a unique array of substrates. One validated DNAJ-PKAc target is the Bcl-2 associated athanogene 2 (BAG2), a co-chaperone recruited to the fusion kinase through association with Hsp70. Immunoblot and immunohistochemical analyses of FLC patient samples correlate increased levels of BAG2 with advanced disease and metastatic recurrences. BAG2 is linked to Bcl-2, an anti-apoptotic factor that delays cell death. Pharmacological approaches tested if the DNAJ- PKAc/Hsp70/BAG2 axis contributes to chemotherapeutic resistance in AML12DNAJ-PKAc hepatocyte cell lines using the DNA damaging agent etoposide and the Bcl-2 inhibitor navitoclax. Wildtype AML12 cells were susceptible to each drug alone and in combination. In contrast, AML12DNAJ-PKAc cells were moderately affected by etoposide, resistant to navitoclax, but markedly susceptible to the drug combination. These studies implicate BAG2 as a biomarker for advanced FLC and a chemotherapeutic resistance factor in DNAJ-PKAc signaling scaffolds.
]]></description>
<dc:creator>Lauer, S. M.-L.</dc:creator>
<dc:creator>Omar, M. H.</dc:creator>
<dc:creator>Golkowski, M. G.</dc:creator>
<dc:creator>Kenerson, H. L.</dc:creator>
<dc:creator>Pascual, B. C.</dc:creator>
<dc:creator>Forbush, K.</dc:creator>
<dc:creator>Smith, F. D.</dc:creator>
<dc:creator>Gordan, J.</dc:creator>
<dc:creator>Ong, S.-E. M.</dc:creator>
<dc:creator>Yeung, R. S.</dc:creator>
<dc:creator>Scott, J. D.</dc:creator>
<dc:date>2023-06-28</dc:date>
<dc:identifier>doi:10.1101/2023.06.28.546958</dc:identifier>
<dc:title><![CDATA[Recruitment of BAG2 to DNAJ-PKAc scaffolds promotes cell survival and resistance to drug-induced apoptosis in fibrolamellar carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.28.546894v1?rss=1">
<title>
<![CDATA[
Combinatorial transcription factor binding encodes cis-regulatory wiring of forebrain GABAergic neurogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.28.546894v1?rss=1</link>
<description><![CDATA[
Transcription factors (TFs) bind combinatorially to genomic cis-regulatory elements (cREs), orchestrating transcription programs. While studies of chromatin state and chromosomal interactions have revealed dynamic neurodevelopmental cRE landscapes, parallel understanding of the underlying TF binding lags. To elucidate the combinatorial TF-cRE interactions driving mouse basal ganglia development, we integrated ChIP-seq for twelve TFs, H3K4me3-associated enhancer-promoter interactions, chromatin and transcriptional state, and transgenic enhancer assays. We identified TF-cREs modules with distinct chromatin features and enhancer activity that have complementary roles driving GABAergic neurogenesis and suppressing other developmental fates. While the majority of distal cREs were bound by one or two TFs, a small proportion were extensively bound, and these enhancers also exhibited exceptional evolutionary conservation, motif density, and complex chromosomal interactions. Our results provide new insights into how modules of combinatorial TF-cRE interactions activate and repress developmental expression programs and demonstrate the value of TF binding data in modeling gene regulatory wiring.
]]></description>
<dc:creator>Catta-Preta, R.</dc:creator>
<dc:creator>Lindtner, S.</dc:creator>
<dc:creator>Ypsilanti, A.</dc:creator>
<dc:creator>Price, J.</dc:creator>
<dc:creator>Abnousi, A.</dc:creator>
<dc:creator>Su-Feher, L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Juric, I.</dc:creator>
<dc:creator>Jones, I.</dc:creator>
<dc:creator>Akiyama, J.</dc:creator>
<dc:creator>Hu, M.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Visel, A.</dc:creator>
<dc:creator>Pennacchio, L.</dc:creator>
<dc:creator>Dickel, D.</dc:creator>
<dc:creator>Rubenstein, J.</dc:creator>
<dc:creator>Nord, A.</dc:creator>
<dc:date>2023-06-29</dc:date>
<dc:identifier>doi:10.1101/2023.06.28.546894</dc:identifier>
<dc:title><![CDATA[Combinatorial transcription factor binding encodes cis-regulatory wiring of forebrain GABAergic neurogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.28.546819v1?rss=1">
<title>
<![CDATA[
Discovery of New Broad-Spectrum Anti-Infectives for Eukaryotic Pathogens Using Bioorganometallic Chemistry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.28.546819v1?rss=1</link>
<description><![CDATA[
Drug resistance observed with many anti-infectives clearly highlights the need for new broad-spectrum agents to treat especially neglected tropical diseases (NTDs) caused by eukaryotic parasitic pathogens including fungal infections. Since these diseases target the most vulnerable communities who are disadvantaged by health and socio-economic factors, new agents should be, if possible, easy-to-prepare to allow for commercialization based on their low cost. In this study, we show that simple modification of one of the most well-known antifungal drugs, fluconazole, with organometallic moieties not only improves the activity of the parent drug but also broadens the scope of application of the new derivatives. These compounds were highly effective in vivo against pathogenic fungal infections and potent against parasitic worms such as Brugia, which causes lymphatic filariasis and Trichuris, one of the soil-transmitted helminths that infects millions of people globally. Notably, the identified molecular targets indicate a mechanism of action that differs greatly from the parental antifungal drug, including targets involved in biosynthetic pathways that are absent in humans, offering great potential to expand our armamentarium against drug-resistant fungal infections and NTDs targeted for elimination by 2030. Overall, the discovery of these new compounds with broad-spectrum activity opens new avenues for the development of treatments for several current human infections, either caused by fungi or by parasites, including other NTDs, as well as newly emerging diseases.

ONE-SENTENCE SUMMARYSimple derivatives of the well-known antifungal drug fluconazole were found to be highly effective in vivo against fungal infections, and also potent against the parasitic nematode Brugia, which causes lymphatic filariasis and against Trichuris, one of the soil-transmitted helminths that infects millions of people globally.
]]></description>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Jung, H.</dc:creator>
<dc:creator>Bulman, C. A.</dc:creator>
<dc:creator>Ng, J.</dc:creator>
<dc:creator>Vinck, R.</dc:creator>
<dc:creator>O'Beirne, C.</dc:creator>
<dc:creator>Moser, M. S.</dc:creator>
<dc:creator>Tricoche, N.</dc:creator>
<dc:creator>Peguero, R.</dc:creator>
<dc:creator>Li, R. W.</dc:creator>
<dc:creator>Urban, J. F.</dc:creator>
<dc:creator>Le Pape, P.</dc:creator>
<dc:creator>Pagniez, F.</dc:creator>
<dc:creator>Moretto, M.</dc:creator>
<dc:creator>Weil, T.</dc:creator>
<dc:creator>Lustigman, S.</dc:creator>
<dc:creator>Cariou, K.</dc:creator>
<dc:creator>Mitreva, M.</dc:creator>
<dc:creator>Sakanari, J.</dc:creator>
<dc:creator>Gasser, G.</dc:creator>
<dc:date>2023-06-30</dc:date>
<dc:identifier>doi:10.1101/2023.06.28.546819</dc:identifier>
<dc:title><![CDATA[Discovery of New Broad-Spectrum Anti-Infectives for Eukaryotic Pathogens Using Bioorganometallic Chemistry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.30.547215v1?rss=1">
<title>
<![CDATA[
Differential susceptibility of male and female germ cells to glucocorticoid-mediated signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.30.547215v1?rss=1</link>
<description><![CDATA[
While physiologic stress has long been known to impair mammalian reproductive capacity through hormonal dysregulation, mounting evidence now suggests that stress experienced prior to or during gestation may also negatively impact the health of future offspring. Rodent models of gestational physiologic stress can induce neurologic and behavioral changes that persist for up to three generations, suggesting that stress signals can induce lasting epigenetic changes in the germline. Treatment with glucocorticoid stress hormones is sufficient to recapitulate the transgenerational changes seen in physiologic stress models. These hormones are known to bind and activate the glucocorticoid receptor (GR), a ligand-inducible transcription factor, thus implicating GR-mediated signaling as a potential contributor to the transgenerational inheritance of stress-induced phenotypes. Here we demonstrate dynamic spatiotemporal regulation of GR expression in the mouse germline, showing expression in the fetal oocyte as well as the perinatal and adult spermatogonia. Functionally, we find that fetal oocytes are intrinsically buffered against changes in GR signaling, as neither genetic deletion of GR nor GR agonism with dexamethasone altered the transcriptional landscape or the progression of fetal oocytes through meiosis. In contrast, our studies revealed that the male germline is susceptible to glucocorticoid-mediated signaling, specifically by regulating RNA splicing within the spermatogonia, although this does not abrogate fertility. Together, our work suggests a sexually dimorphic function for GR in the germline, and represents an important step towards understanding the mechanisms by which stress can modulate the transmission of genetic information through the germline.
]]></description>
<dc:creator>Cincotta, S. A.</dc:creator>
<dc:creator>Richardson, N.</dc:creator>
<dc:creator>Foecke, M. H.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:date>2023-07-01</dc:date>
<dc:identifier>doi:10.1101/2023.06.30.547215</dc:identifier>
<dc:title><![CDATA[Differential susceptibility of male and female germ cells to glucocorticoid-mediated signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.01.547335v1?rss=1">
<title>
<![CDATA[
Single Cell Multi-Omics of an iPSC Model of Human Sinoatrial Node Development Reveals Genetic Determinants of Heart Rate and Arrhythmia Susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.01.547335v1?rss=1</link>
<description><![CDATA[
Cellular heterogeneity within the sinoatrial node (SAN) is functionally important but has been difficult to model in vitro, presenting a major obstacle to studies of heart rate regulation and arrhythmias. Here we describe a scalable method to derive sinoatrial node pacemaker cardiomyocytes (PCs) from human induced pluripotent stem cells that recapitulates differentiation into distinct PC subtypes, including SAN Head, SAN Tail, transitional zone cells, and sinus venosus myocardium. Single cell (sc) RNA-sequencing, sc-ATAC-sequencing, and trajectory analyses were used to define epigenetic and transcriptomic signatures of each cell type, and to identify novel transcriptional pathways important for PC subtype differentiation. Integration of our multi-omics datasets with genome wide association studies uncovered cell type-specific regulatory elements that associated with heart rate regulation and susceptibility to atrial fibrillation. Taken together, these datasets validate a novel, robust, and realistic in vitro platform that will enable deeper mechanistic exploration of human cardiac automaticity and arrhythmia.
]]></description>
<dc:creator>Engel, J. L.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Lu, D. R.</dc:creator>
<dc:creator>Vila, O. F.</dc:creator>
<dc:creator>Arias, V.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Hale, C.</dc:creator>
<dc:creator>Hsu, Y.-H.</dc:creator>
<dc:creator>Li, C.-M. K.</dc:creator>
<dc:creator>Vedantham, V.</dc:creator>
<dc:creator>Ang, Y.-S.</dc:creator>
<dc:date>2023-07-01</dc:date>
<dc:identifier>doi:10.1101/2023.07.01.547335</dc:identifier>
<dc:title><![CDATA[Single Cell Multi-Omics of an iPSC Model of Human Sinoatrial Node Development Reveals Genetic Determinants of Heart Rate and Arrhythmia Susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.01.546933v1?rss=1">
<title>
<![CDATA[
Configurational entropy is an intrinsic driver of tissue structural heterogeneity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.01.546933v1?rss=1</link>
<description><![CDATA[
Tissues comprise ordered arrangements of cells that can be surprisingly disordered in their details. How the properties of single cells and their microenvironment contribute to the balance between order and disorder at the tissue-scale remains poorly understood. Here, we address this question using the self-organization of human mammary organoids as a model. We find that organoids behave like a dynamic structural ensemble at the steady state. We apply a maximum entropy formalism to derive the ensemble distribution from three measurable parameters - the degeneracy of structural states, interfacial energy, and tissue activity (the energy associated with positional fluctuations). We link these parameters with the molecular and microenvironmental factors that control them to precisely engineer the ensemble across multiple conditions. Our analysis reveals that the entropy associated with structural degeneracy sets a theoretical limit to tissue order and provides new insight for tissue engineering, development, and our understanding of disease progression.
]]></description>
<dc:creator>Srivastava, V.</dc:creator>
<dc:creator>Hu, J. L.</dc:creator>
<dc:creator>Garbe, J. C.</dc:creator>
<dc:creator>Veytsman, B.</dc:creator>
<dc:creator>Shalabi, S. F.</dc:creator>
<dc:creator>Yllanes, D.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:creator>LaBarge, M. A.</dc:creator>
<dc:creator>Huber, G.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:date>2023-07-02</dc:date>
<dc:identifier>doi:10.1101/2023.07.01.546933</dc:identifier>
<dc:title><![CDATA[Configurational entropy is an intrinsic driver of tissue structural heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.01.547331v1?rss=1">
<title>
<![CDATA[
VTA dopamine neurons drive spatiotemporally heterogeneous striatal dopamine signals during learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.01.547331v1?rss=1</link>
<description><![CDATA[
Environmental cues, through Pavlovian learning, become conditioned stimuli that invigorate and guide animals toward acquisition of rewards. Dopamine neurons in the ventral tegmental area (VTA) and substantia nigra (SNC) are crucial for this process. Dopamine neurons are embedded in a reciprocally connected network with their striatal targets, the functional organization of which remains poorly understood. Here, we investigated how learning during optogenetic Pavlovian cue conditioning of VTA or SNC dopamine neurons directs cue-evoked behavior and shapes subregion-specific striatal dopamine dynamics. We used a fluorescent dopamine biosensor to monitor dopamine in the nucleus accumbens (NAc) core and shell, dorsomedial striatum (DMS), and dorsolateral striatum (DLS). We demonstrate spatially heterogeneous, learning-dependent dopamine changes across striatal regions. While VTA stimulation evoked robust dopamine release in NAc core, shell, and DMS, cues predictive of this activation preferentially recruited dopamine release in NAc core, starting early in training, and DMS, late in training. Corresponding negative prediction error signals, reflecting a violation in the expectation of dopamine neuron activation, only emerged in the NAc core and DMS, and not the shell. Despite development of vigorous movement late in training, conditioned dopamine signals did not similarly emerge in the DLS, even during Pavlovian conditioning with SNC dopamine neuron activation, which elicited robust DLS dopamine release. Together, our studies show broad dissociation in the fundamental prediction and reward-related information generated by different dopamine neuron populations and signaled by dopamine across the striatum. Further, they offer new insight into how larger-scale plasticity across the striatal network emerges during Pavlovian learning to coordinate behavior.
]]></description>
<dc:creator>Engel, L.</dc:creator>
<dc:creator>Wolff, A. R.</dc:creator>
<dc:creator>Blake, M.</dc:creator>
<dc:creator>Collins, V. L.</dc:creator>
<dc:creator>Sinha, S.</dc:creator>
<dc:creator>Saunders, B. T.</dc:creator>
<dc:date>2023-07-02</dc:date>
<dc:identifier>doi:10.1101/2023.07.01.547331</dc:identifier>
<dc:title><![CDATA[VTA dopamine neurons drive spatiotemporally heterogeneous striatal dopamine signals during learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.01.547359v1?rss=1">
<title>
<![CDATA[
Chemical alkylation of Asp12 enables mutant selective targeting of K-Ras(G12D) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.01.547359v1?rss=1</link>
<description><![CDATA[
K-Ras is the most commonly mutated oncogene in human cancer, yet direct small-molecule targeting of K-Ras mutants has been mostly unsuccessful until recently. The discovery of an allosteric pocket under Switch-II with covalent cysteine-crosslinking molecules has allowed for the development of targeted therapies that selectively engage the highly reactive acquired cysteine in the K-Ras(G12C) mutation without affecting the wild-type protein. Sotorasib and adagrasib, two advanced Switch-II Pocket inhibitors, have received FDA approval to treat K-Ras(G12C)-driven non-small cell lung cancer. However, the most frequent K-Ras mutation G12D particularly prevalent in pancreatic ductal adenocarcinoma has remained untargetable with covalent drugs due to the poor nucleophilicity of the somatic aspartate residue. Here we present a set of malolactone-based electrophiles which exploit ring strain to crosslink K-Ras(G12D) at the mutant aspartate to form stable covalent complexes. Structural insights from x-ray crystallography and exploitation of the stereoelectronic requirements for attack of the electrophile allowed development of a substituted malolactone which resisted attack by aqueous buffer but rapidly crosslink with the aspartate-12 of K-Ras in both GDP- and GTP-state. The signaling-competent GTP-state targeting allowed effective suppression of downstream signaling and proliferation of cancer cells harboring K-Ras(G12D) mutation, and tumor growth of cell line-derive xenograft in mice. Our results demonstrate the rational design of covalent inhibitors to target a non-catalytic carboxylic acid side chain in K-Ras(G12D) which has resisted traditional drug discovery efforts.
]]></description>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Zheng, Q.</dc:creator>
<dc:creator>Guiley, K. Z.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:date>2023-07-02</dc:date>
<dc:identifier>doi:10.1101/2023.07.01.547359</dc:identifier>
<dc:title><![CDATA[Chemical alkylation of Asp12 enables mutant selective targeting of K-Ras(G12D)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.30.547223v1?rss=1">
<title>
<![CDATA[
Inference for entomological semi-field experiments: Fitting a mathematical model assessing personal and community protection of vector-control interventions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.30.547223v1?rss=1</link>
<description><![CDATA[
The effectiveness of vector-control tools is often assessed by experiments as a reduction in mosquito landings using human landing catches (HLCs). However, HLCs alone only quantify a single characteristic and therefore do not provide information on the overall impacts of the intervention product. Using data from a recent semi-field study which used time-stratified HLCs, aspiration of non-landing mosquitoes, and blood feeding, we suggest a Bayesian inference approach for fitting such data to a stochastic model. This model considers both personal protection, through a reduction in biting, and community protection, from mosquito mortality and disarming (prolonged inhibition of blood feeding). Parameter estimates are then used to predict the reduction of vectorial capacity induced by etofenpox-treated clothing, picaridin topical repellents, transfluthrin spatial repellents and metofluthrin spatial repellents, as well as combined interventions for Plasmodium falciparum malaria in Anopleles minimus. Overall, all interventions had both personal and community effects, preventing biting and killing or disarming mosquitoes. This led to large estimated reductions in the vectorial capacity, with substantial impact even at low coverage. As the interventions aged, fewer mosquitoes were killed; however the impact of some interventions changed from killing to disarming mosquitoes. Overall, this inference method allows for additional modes of action, rather than just reduction in biting, to be parameterised and highlights the tools assessed as promising malaria interventions.
]]></description>
<dc:creator>Fairbanks, E. L.</dc:creator>
<dc:creator>Saeung, M.</dc:creator>
<dc:creator>Pongsiri, A.</dc:creator>
<dc:creator>Vajda, E.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>McIver, D. J.</dc:creator>
<dc:creator>Tatarsky, A.</dc:creator>
<dc:creator>Lobo, N. F.</dc:creator>
<dc:creator>Moore, S. J.</dc:creator>
<dc:creator>Ponlawat, A.</dc:creator>
<dc:creator>Chareonviriyaphap, T.</dc:creator>
<dc:creator>Ross, A.</dc:creator>
<dc:creator>Chitnis, N.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.06.30.547223</dc:identifier>
<dc:title><![CDATA[Inference for entomological semi-field experiments: Fitting a mathematical model assessing personal and community protection of vector-control interventions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.01.547283v1?rss=1">
<title>
<![CDATA[
KRAS-mediated upregulation of CIP2A promotes suppression of PP2A-B56α to initiate pancreatic cancer development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.01.547283v1?rss=1</link>
<description><![CDATA[
Oncogenic mutations in KRAS are present in approximately 95% of patients diagnosed with pancreatic ductal adenocarcinoma (PDAC) and are considered the initiating event of pancreatic intraepithelial neoplasia (PanIN) precursor lesions. While it is well established that KRAS mutations drive the activation of oncogenic kinase cascades during pancreatic oncogenesis, the effects of oncogenic KRAS signaling on regulation of phosphatases during this process is not fully appreciated. Protein Phosphatase 2A (PP2A) has been implicated in suppressing KRAS-driven cellular transformation. However, low PP2A activity is observed in PDAC cells compared to non-transformed cells, suggesting that suppression of PP2A activity is an important step in the overall development of PDAC. In the current study, we demonstrate that KRASG12D induces the expression of both an endogenous inhibitor of PP2A activity, Cancerous Inhibitor of PP2A (CIP2A), and the PP2A substrate, c-MYC. Consistent with these findings, KRASG12D sequestered the specific PP2A subunit responsible for c-MYC degradation, B56, away from the active PP2A holoenzyme in a CIP2A-dependent manner. During PDAC initiation in vivo, knockout of B56 promoted KRASG12D tumorigenesis by accelerating acinar-to-ductal metaplasia (ADM) and the formation of PanIN lesions. The process of ADM was attenuated ex vivo in response to pharmacological re-activation of PP2A utilizing direct small molecule activators of PP2A (SMAPs). Together, our results suggest that suppression of PP2A-B56 through KRAS signaling can promote the MYC-driven initiation of pancreatic tumorigenesis.
]]></description>
<dc:creator>Tinsley, S. L.</dc:creator>
<dc:creator>Shelley, R. A.</dc:creator>
<dc:creator>Mall, G. K.</dc:creator>
<dc:creator>Chianis, E. R. D.</dc:creator>
<dc:creator>Dhiman, A.</dc:creator>
<dc:creator>Baral, G.</dc:creator>
<dc:creator>Kothandaraman, H.</dc:creator>
<dc:creator>Thoma, M. C.</dc:creator>
<dc:creator>Daniel, C.</dc:creator>
<dc:creator>Lanman, N. A.</dc:creator>
<dc:creator>Pasca di Magliano, M.</dc:creator>
<dc:creator>Narla, G.</dc:creator>
<dc:creator>Solorio, L.</dc:creator>
<dc:creator>Dykhuizen, E.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Allen-Petersen, B.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.01.547283</dc:identifier>
<dc:title><![CDATA[KRAS-mediated upregulation of CIP2A promotes suppression of PP2A-B56α to initiate pancreatic cancer development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.03.547434v1?rss=1">
<title>
<![CDATA[
Solute Transport in Engineered Living Materials using Bone Inspired Microscale Channel Networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.03.547434v1?rss=1</link>
<description><![CDATA[
Engineered living materials (ELMs) are an emerging class of materials that is synthesized and/or populated by living cells. Maintaining living cells within an ELM over prolonged periods remains a major technical challenge that limits the service life of a material. Biological materials regularly maintain robust populations of living cells. Bone maintains living cells for decades by delivering nutrients through a network of nanoscale channels punctuated by microscale pores. Nutrient transfer in bone is enabled by mechanical loading experienced during regular use. Here we identify the characteristics of channel-pore network geometries and external mechanical loading that can be used in engineered living materials to deliver nutrients to resident cell populations. Transport occurs when deformation in the microscale pore network exceeds the volume of the connecting channels. Computational models show that transport is enhanced at greater load magnitudes and lower loading frequencies and are consistent with experimental validation using microfluidic systems. Our findings provide quantitative design principles for channel-pore networks capable of delivering nutrients to materials designed to house living cells.
]]></description>
<dc:creator>van Wijngaarden, E. W.</dc:creator>
<dc:creator>Bratcher, S.</dc:creator>
<dc:creator>Lewis, K. J.</dc:creator>
<dc:creator>Hernandez, C. J.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.03.547434</dc:identifier>
<dc:title><![CDATA[Solute Transport in Engineered Living Materials using Bone Inspired Microscale Channel Networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.03.547549v1?rss=1">
<title>
<![CDATA[
Activity-Induced MeCP2 Phosphorylation Regulates Retinogeniculate Synapse Refinement 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.03.547549v1?rss=1</link>
<description><![CDATA[
Mutations in MECP2 give rise to Rett syndrome (RTT), an X-linked neurodevelopmental disorder that results in broad cognitive impairments in females. While the exact etiology of RTT symptoms remains unknown, one possible explanation for its clinical presentation is that loss of MeCP2 causes miswiring of neural circuits due to defects in the brains capacity to respond to changes in neuronal activity and sensory experience. Here we show that MeCP2 is phosphorylated at four residues in the brain (S86, S274, T308, and S421) in response to neuronal activity, and we generate a quadruple knock-in (QKI) mouse line in which all four activity-dependent sites are mutated to alanines to prevent phosphorylation. QKI mice do not display overt RTT phenotypes or detectable gene expression changes in two brain regions. However, electrophysiological recordings from the retinogeniculate synapse of QKI mice reveal that while synapse elimination is initially normal at P14, it is significantly compromised at P20. Notably, this phenotype is distinct from that previously reported for Mecp2 null mice, where synapses initially refine but then regress after the third postnatal week. We thus propose a model in which activity-induced phosphorylation of MeCP2 is critical for the proper timing of retinogeniculate synapse maturation specifically during the early postnatal period.

SIGNIFICANCE STATEMENTRett syndrome (RTT) is an X-linked neurodevelopmental disorder that predominantly affects girls. RTT is caused by loss of function mutations in a single gene MeCP2. Girls with RTT develop normally during their first year of life, but then experience neurological abnormalities including breathing and movement difficulties, loss of speech, and seizures. This study investigates the function of the MeCP2 protein in the brain, and how MeCP2 activity is modulated by sensory experience in early life. Evidence is presented that sensory experience affects MeCP2 function, and that this is required for synaptic pruning in the brain. These findings provide insight into MeCP2 function, and clues as to what goes awry in the brain when the function of MeCP2 is disrupted.
]]></description>
<dc:creator>Tzeng, C. P.</dc:creator>
<dc:creator>Whitwam, T.</dc:creator>
<dc:creator>Boxer, L. D.</dc:creator>
<dc:creator>Li, E.</dc:creator>
<dc:creator>Silberfeld, A.</dc:creator>
<dc:creator>Trowbridge, S.</dc:creator>
<dc:creator>Mei, K.</dc:creator>
<dc:creator>Lin, C.</dc:creator>
<dc:creator>Shamah, R.</dc:creator>
<dc:creator>Griffith, E. C.</dc:creator>
<dc:creator>Renthal, W.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Greenberg, M. E.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.03.547549</dc:identifier>
<dc:title><![CDATA[Activity-Induced MeCP2 Phosphorylation Regulates Retinogeniculate Synapse Refinement]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.04.547736v1?rss=1">
<title>
<![CDATA[
Cryo-EM structure of TMEM63C suggests it functions as a monomer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.04.547736v1?rss=1</link>
<description><![CDATA[
The TMEM63 family proteins (A, B, and C), calcium-permeable channels in animals that are preferentially activated by hypo-osmolality, have been implicated in various physiological functions. Deficiency of these channels would cause many diseases including hearing loss. However, their structures and physiological roles are not yet well understood. In this study, we determined the cryo-EM structure of the mouse TMEM63C at 3.56 [A], and revealed structural differences compared to its plant orthologues OSCAs. Further structural guided mutagenesis and electrophysiological studies demonstrated the important roles of the coupling of TM0 and TM6 in channel activity. Additionally, we confirmed that the physiological state of TMEM63C is monomer, while TMEM63B is a mix of monomer and dimer in cells, suggesting that oligomerization is a novel regulatory mechanism for TMEM63 proteins.
]]></description>
<dc:creator>Qin, Y.</dc:creator>
<dc:creator>Yu, D.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Dong, J.</dc:creator>
<dc:creator>Li, W. T.</dc:creator>
<dc:creator>Ye, C.</dc:creator>
<dc:creator>Cheung, K. C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Shi, Y. S.</dc:creator>
<dc:creator>Dang, S.</dc:creator>
<dc:date>2023-07-04</dc:date>
<dc:identifier>doi:10.1101/2023.07.04.547736</dc:identifier>
<dc:title><![CDATA[Cryo-EM structure of TMEM63C suggests it functions as a monomer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.05.547774v1?rss=1">
<title>
<![CDATA[
Recyclable CRISPR/Cas9 mediated gene disruption and deletions in Histoplasma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.05.547774v1?rss=1</link>
<description><![CDATA[
Targeted gene disruption is challenging in the dimorphic fungal pathogen Histoplasma due to the low frequency of homologous recombination. Transformed DNA is either integrated ectopically into the genome or maintained extra chromosomally by de novo addition of telomeric sequences. Based on a system developed in Blastomyces, we adapted a CRISPR/Cas9 system to facilitate targeted gene disruption in Histoplasma with high efficiency. We express a codon-optimized version of Cas9 as well as guide RNAs from a single ectopic vector carrying a selectable marker. Once the desired mutation is verified, one can screen for isolates that have lost the Cas9 vector by simply removing the selective pressure. Multiple mutations can then be generated in the same strain by retransforming the Cas9 vector carrying different guides. We used this system to disrupt a number of target genes including RYP2 and SRE1 where loss-of-function mutations could be monitored visually by colony morphology or color, respectively. Interestingly, expression of two guide RNAs targeting the 5 and 3 ends of a gene allowed isolation of deletion mutants where the sequence between the guide RNAs was removed from the genome. Whole-genome sequencing showed that the frequency of off-target mutations associated with the Cas9 nuclease was negligible. Finally, we increased the frequency of gene disruption by using an endogenous Histoplasma regulatory sequence to drive guide RNA expression. These tools transform our ability to generate targeted mutations in Histoplasma.

ImportanceHistoplasma is a primary fungal pathogen with the ability to infect otherwise healthy mammalian hosts, causing systemic and sometimes life-threatening disease. Thus far, molecular genetic manipulation of this organism has utilized RNA interference, random insertional mutagenesis, and a homologous recombination protocol that is highly variable and often inefficient. Targeted gene manipulations have been challenging due to poor rates of homologous recombination events in Histoplasma. Interrogation of the virulence strategies of this organism would be highly accelerated by a means of efficiently generating targeted mutations. We have developed a recyclable CRISPR/Cas9 system that can be used to introduce gene disruptions in Histoplasma with high efficiency, thereby allowing disruption of multiple genes.
]]></description>
<dc:creator>Joehnk, B.</dc:creator>
<dc:creator>Ali, N.</dc:creator>
<dc:creator>Voorhies, M.</dc:creator>
<dc:creator>Walcott, K.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:date>2023-07-05</dc:date>
<dc:identifier>doi:10.1101/2023.07.05.547774</dc:identifier>
<dc:title><![CDATA[Recyclable CRISPR/Cas9 mediated gene disruption and deletions in Histoplasma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.05.547902v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 Nsp1 regulates translation start site fidelity to promote infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.05.547902v1?rss=1</link>
<description><![CDATA[
A better mechanistic understanding of virus-host interactions can help reveal vulnerabilities and identify opportunities for therapeutic interventions. Of particular interest are essential interactions that enable production of viral proteins, as those could target an early step in the virus lifecycle. Here, we use subcellular proteomics, ribosome profiling analyses and reporter assays to detect changes in polysome composition and protein synthesis during SARS-CoV-2 (CoV2) infection. We identify specific translation factors and molecular chaperones whose inhibition impairs infectious particle production without major toxicity to the host. We find that CoV2 non-structural protein Nsp1 selectively enhances virus translation through functional interactions with initiation factor EIF1A. When EIF1A is depleted, more ribosomes initiate translation from an upstream CUG start codon, inhibiting translation of non-structural genes and reducing viral titers. Together, our work describes multiple dependencies of CoV2 on host biosynthetic networks and identifies druggable targets for potential antiviral development.
]]></description>
<dc:creator>Aviner, R.</dc:creator>
<dc:creator>Lidsky, P. V.</dc:creator>
<dc:creator>Xiao, Y.</dc:creator>
<dc:creator>Tasseto, M.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>McAlpine, P. L.</dc:creator>
<dc:creator>Elias, J.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:date>2023-07-07</dc:date>
<dc:identifier>doi:10.1101/2023.07.05.547902</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 Nsp1 regulates translation start site fidelity to promote infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.06.547567v1?rss=1">
<title>
<![CDATA[
Mapping general anesthesia states based on electro-encephalogram transition phases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.06.547567v1?rss=1</link>
<description><![CDATA[
Cortical electro-encephalography (EEG) has become the clinical reference for monitoring unconsciousness during general anesthesia. The current EEG-based monitors classify general anesthesia states simply as underdosed, adequate, or overdosed, with no transition phases among these states, and therefore no predictive power. To address the issue of transition phases, we analyzed EEG signal of isoflurane-induced general anesthesia in mice. We adopted a data-driven approach and utilized signal processing to track{theta} - and{delta} - band dynamics as well as iso-electric suppressions. By combining this approach with machine learning, we developed a fully-automated algorithm. We found that the dampening of the{delta} -band occurred several minutes before significant iso-electric suppression episodes. Additionally, we observed a distinct{gamma} -frequency oscillation that persisted for several minutes during the recovery phase following isoflurane-induced overdose. Finally, we constructed a map summarizing multiple states and their transitions which can be utilized to predict and prevent overdose during general anesthesia. The transition phases we identified and algorithm we developed may allow clinicians to prevent inadequate anesthesia, and thus individually tailor anesthetic regimens.

1 Significance statementIn human patients, overdosing during general anesthesia can lead to cognitive impairment. Cortical electro-encephalograms are used to measure the depth of anesthesia. They allow for correction, but not prevention, of overdose. However, data-driven approaches open new possibilities to predict the depth of anesthesia. We established an electro-encephalogram signalprocessing pipeline, and constructed a predictive map representing an ensemble of gradual sedation states during general anesthesia in mice. In particular, we identified key electroencephalogram patterns which anticipate signs of overdose several minutes before they occur. Our results bring a novel paradigm to the medical community, allowing for the development of individually tailored and predictive anesthetic regimens.
]]></description>
<dc:creator>Loison, V.</dc:creator>
<dc:creator>Voskobiynyk, Y.</dc:creator>
<dc:creator>Lindquist, B.</dc:creator>
<dc:creator>Necula, D.</dc:creator>
<dc:creator>Longrois, D.</dc:creator>
<dc:creator>Paz, J. T.</dc:creator>
<dc:creator>Holcman, D.</dc:creator>
<dc:date>2023-07-07</dc:date>
<dc:identifier>doi:10.1101/2023.07.06.547567</dc:identifier>
<dc:title><![CDATA[Mapping general anesthesia states based on electro-encephalogram transition phases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.06.548027v1?rss=1">
<title>
<![CDATA[
A meta-analysis of mouse models of tauopathy reveals the cellular underpinnings of selective regional vulnerability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.06.548027v1?rss=1</link>
<description><![CDATA[
Neurodegenerative diseases such as Alzheimers disease (AD) exhibit pathological changes in the brain that proceed in a stereotyped and regionally specific fashion, but the cellular and molecular underpinnings of regional vulnerability are currently poorly understood. Recent work has identified certain subpopulations of neurons in a few focal regions of interest, such as the entorhinal cortex, that are selectively vulnerable to tau pathology in AD. However, the cellular underpinnings of regional susceptibility to tau pathology are currently unknown, primarily because whole-brain maps of a comprehensive collection of cell types have been inaccessible. Here, we deployed a recent cell-type mapping pipeline, Matrix Inversion and Subset Selection (MISS), to determine the brain-wide distributions of pan-hippocampal and neocortical neuronal and non-neuronal cells in the mouse using recently available single-cell RNA sequencing (scRNAseq) data. We then performed a robust set of analyses to identify general principles of cell-type-based selective vulnerability using these cell-type distributions, utilizing 5 transgenic mouse studies that quantified regional tau in 12 distinct PS19 mouse models. Using our approach, which constitutes the broadest exploration of whole-brain selective vulnerability to date, we were able to discover cell types and cell-type classes that conferred vulnerability and resilience to tau pathology. Hippocampal glutamatergic neurons as a whole were strongly positively associated with regional tau deposition, suggesting vulnerability, while cortical glutamatergic and GABAergic neurons were negatively associated. Among glia, we identified oligodendrocytes as the single-most strongly negatively associated cell type, whereas microglia were consistently positively correlated. Strikingly, we found that there was no association between the gene expression relationships between cell types and their vulnerability or resilience to tau pathology. When we looked at the explanatory power of cell types versus GWAS-identified AD risk genes, cell type distributions were consistently more predictive of end-timepoint tau pathology than regional gene expression. To understand the functional enrichment patterns of the genes that were markers of the identified vulnerable or resilient cell types, we performed gene ontology analysis. We found that the genes that are directly correlated to tau pathology are functionally distinct from those that constitutively embody the vulnerable cells. In short, we have demonstrated that regional cell-type composition is a compelling explanation for the selective vulnerability observed in tauopathic diseases at a whole-brain level and is distinct from that conferred by risk genes. These findings may have implications in identifying cell-type-based therapeutic targets.
]]></description>
<dc:creator>Torok, J.</dc:creator>
<dc:creator>Maia, P.</dc:creator>
<dc:creator>Anand, C.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:date>2023-07-07</dc:date>
<dc:identifier>doi:10.1101/2023.07.06.548027</dc:identifier>
<dc:title><![CDATA[A meta-analysis of mouse models of tauopathy reveals the cellular underpinnings of selective regional vulnerability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.06.548037v1?rss=1">
<title>
<![CDATA[
Using Published Pathway Figures in Enrichment Analysis and Machine Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.06.548037v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWPathway Figure OCR (PFOCR) is a novel kind of pathway database approaching the breadth and depth of Gene Ontology while providing rich, mechanistic diagrams and direct literature support. PFOCR content is extracted from published pathway figures currently emerging at a rate of 1000 new pathways each month. Here, we compare the pathway information contained in PFOCR against popular pathway databases with respect to overall and disease-specific coverage. In addition to common pathways analysis use cases, we present two advanced case studies demonstrating unique advantages of PFOCR in terms of cancer subtype and grade prediction analyses.
]]></description>
<dc:creator>SHIN, M.-G.</dc:creator>
<dc:creator>Pico, A.</dc:creator>
<dc:date>2023-07-07</dc:date>
<dc:identifier>doi:10.1101/2023.07.06.548037</dc:identifier>
<dc:title><![CDATA[Using Published Pathway Figures in Enrichment Analysis and Machine Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.09.548214v1?rss=1">
<title>
<![CDATA[
Tetracycline transactivator overexpression in keratinocytes triggers a TRPV1 primary sensory neuron-dependent neuropathic itch 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.09.548214v1?rss=1</link>
<description><![CDATA[
Mouse models that combine tetracycline-controlled gene expression systems and conditional genetic activation can tightly regulate transgene expression in discrete cell types and tissues. However, the commonly used Tet-Off variant, tetracycline transactivator (tTA), when overexpressed and fully active, can lead to developmental lethality, disease, or more subtle behavioral phenotypes. Here we describe a profound itch phenotype in mice expressing a genetically encoded tTA that is conditionally activated within the Phox2a lineage. Phox2a; tTA mice develop intense, localized scratching and regional skin lesions that can be controlled by the tTA inhibitor, doxycycline. As gabapentin, but not morphine, relieved the scratching, we consider this phenotype to result from chronic neuropathic itch, not pain. In contrast to the Phox2a lineage, mice with tTA activated within the Phox2b lineage, which has many similar areas of recombination within the nervous system, did not recapitulate the scratching phenotype. In Phox2a-Cre mice, but not Phox2b-Cre, intense Cre-dependent reporter expression was found in skin keratinocytes which formed the area at which skin lesions developed. Most interestingly, repeated topical application of the DREADD agonist, CNO, which chronically induced Gi signaling in Phox2a-keratinocytes, completely reversed the localized scratching and skin lesions. Furthermore, ablation of TRPV1-expressing, primary afferent neurons reduced the scratching with a time course comparable to that produced by Gi-DREADD inhibition. These temporal properties suggest that the neuropathic itch condition arises not only from localized keratinocyte activation of peripheral nerves but also from a persistent, gabapentin-sensitive state of central sensitization.
]]></description>
<dc:creator>Crowther, A.</dc:creator>
<dc:creator>Kashem, S.</dc:creator>
<dc:creator>Jewell, M. E.</dc:creator>
<dc:creator>Chang, H. L.</dc:creator>
<dc:creator>Casillas, M. R.</dc:creator>
<dc:creator>Midavaine, E.</dc:creator>
<dc:creator>Rodriguez, S.</dc:creator>
<dc:creator>Braz, J.</dc:creator>
<dc:creator>Kania, A.</dc:creator>
<dc:creator>Basbaum, A.</dc:creator>
<dc:date>2023-07-10</dc:date>
<dc:identifier>doi:10.1101/2023.07.09.548214</dc:identifier>
<dc:title><![CDATA[Tetracycline transactivator overexpression in keratinocytes triggers a TRPV1 primary sensory neuron-dependent neuropathic itch]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.10.548456v1?rss=1">
<title>
<![CDATA[
NOTCH3 drives meningioma tumorigenesis and resistance to radiotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.10.548456v1?rss=1</link>
<description><![CDATA[
Meningiomas are the most common primary intracranial tumors1-3. Treatments for patients with meningiomas are limited to surgery and radiotherapy, and systemic therapies remain ineffective or experimental4,5. Resistance to radiotherapy is common in high-grade meningiomas6, and the cell types and signaling mechanisms driving meningioma tumorigenesis or resistance to radiotherapy are incompletely understood. Here we report NOTCH3 drives meningioma tumorigenesis and resistance to radiotherapy and find NOTCH3+ meningioma mural cells are conserved across meningiomas from humans, dogs, and mice. NOTCH3+ cells are restricted to the perivascular niche during meningeal development and homeostasis and in low-grade meningiomas but are expressed throughout high-grade meningiomas that are resistant to radiotherapy. Integrating single-cell transcriptomics with lineage tracing and imaging approaches across mouse genetic and xenograft models, we show NOTCH3 drives tumor initiating capacity, cell proliferation, angiogenesis, and resistance to radiotherapy to increase meningioma growth and reduce survival. An antibody stabilizing the extracellular negative regulatory region of NOTCH37,8 blocks meningioma tumorigenesis and sensitizes meningiomas to radiotherapy, reducing tumor growth and improving survival in preclinical models. In summary, our results identify a conserved cell type and signaling mechanism that underlie meningioma tumorigenesis and resistance to radiotherapy, revealing a new therapeutic vulnerability to treat meningiomas that are resistant to standard interventions.
]]></description>
<dc:creator>Choudhury, A.</dc:creator>
<dc:creator>Cady, M. A.</dc:creator>
<dc:creator>Lucas, C.-H. G.</dc:creator>
<dc:creator>Najem, H.</dc:creator>
<dc:creator>Phillips, J. J.</dc:creator>
<dc:creator>Palikuqi, B.</dc:creator>
<dc:creator>Zakimi, N.</dc:creator>
<dc:creator>Joseph, T.</dc:creator>
<dc:creator>Birreuta, J. O.</dc:creator>
<dc:creator>Chen, W. C.</dc:creator>
<dc:creator>Bush, N. A. O.</dc:creator>
<dc:creator>Hervey-Jumper, S. L.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Toedebusch, C. M.</dc:creator>
<dc:creator>Horbinski, C. M.</dc:creator>
<dc:creator>Magill, S. T.</dc:creator>
<dc:creator>Bhaduri, A.</dc:creator>
<dc:creator>Perry, A.</dc:creator>
<dc:creator>Dickinson, P. J.</dc:creator>
<dc:creator>Heimberger, A. B.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Crouch, E. E.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:date>2023-07-11</dc:date>
<dc:identifier>doi:10.1101/2023.07.10.548456</dc:identifier>
<dc:title><![CDATA[NOTCH3 drives meningioma tumorigenesis and resistance to radiotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.11.548615v1?rss=1">
<title>
<![CDATA[
Resolving Non-identifiability Mitigates Bias in Models of Neural Tuning and Functional Coupling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.11.548615v1?rss=1</link>
<description><![CDATA[
In the brain, all neurons are driven by the activity of other neurons, some of which maybe simultaneously recorded, but most are not. As such, models of neuronal activity need to account for simultaneously recorded neurons and the influences of unmeasured neurons. This can be done through inclusion of model terms for observed external variables (e.g., tuning to stimuli) as well as terms for latent sources of variability. Determining the influence of groups of neurons on each other relative to other influences is important to understand brain functioning. The parameters of statistical models fit to data are commonly used to gain insight into the relative importance of those influences. Scientific interpretation of models hinge upon unbiased parameter estimates. However, evaluation of biased inference is rarely performed and sources of bias are poorly understood. Through extensive numerical study and analytic calculation, we show that common inference procedures and models are typically biased. We demonstrate that accurate parameter selection before estimation resolves model non-identifiability and mitigates bias. In diverse neurophysiology data sets, we found that contributions of coupling to other neurons are often overestimated while tuning to exogenous variables are underestimated in common methods. We explain heterogeneity in observed biases across data sets in terms of data statistics. Finally, counter to common intuition, we found that model non-identifiability contributes to bias, not variance, making it a particularly insidious form of statistical error. Together, our results identify the causes of statistical biases in common models of neural data, provide inference procedures to mitigate that bias, and reveal and explain the impact of those biases in diverse neural data sets.

1 Author SummaryExperimental data of interacting cells under the influence of external as well as unobserved factors are ubiquitous. Parametric models are often used to gain understanding of the processes that generated such data. As such, biological understanding hinges upon accurate inference of model parameters. Whether and how systemic parameter bias manifests in such models is poorly understood. We study this issue in the specific context of estimating the static and dynamic interactions of simultaneously recorded neurons influenced by stimuli and unobserved neurons. Through extensive numerical study and analytic calculations, we identify and mitigate bias in such models. When applied to diverse neural data sets, we found that common models and inference procedures often overestimate the importance of coupling and underestimate tuning. In contrast to common intuition, we find that model non-identifiability contributes to estimation bias, not variance, making it a particularly insidious form of statistical error. As the experimental and statistical issues examined here are common, the insights and solutions we developed will likely impact many fields of biology.
]]></description>
<dc:creator>Sachdeva, P.</dc:creator>
<dc:creator>Bak, J. H.</dc:creator>
<dc:creator>Livezey, J. A.</dc:creator>
<dc:creator>Frank, L.</dc:creator>
<dc:creator>Kirst, C.</dc:creator>
<dc:creator>Bhattacharyya, S.</dc:creator>
<dc:creator>Bouchard, K.</dc:creator>
<dc:date>2023-07-12</dc:date>
<dc:identifier>doi:10.1101/2023.07.11.548615</dc:identifier>
<dc:title><![CDATA[Resolving Non-identifiability Mitigates Bias in Models of Neural Tuning and Functional Coupling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.12.548736v1?rss=1">
<title>
<![CDATA[
High throughput PRIME editing screens identify functional DNA variants in the human genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.12.548736v1?rss=1</link>
<description><![CDATA[
Despite tremendous progress in detecting DNA variants associated with human disease, interpreting their functional impact in a high-throughput and base-pair resolution manner remains challenging. Here, we develop a novel pooled prime editing screen method, PRIME, which can be applied to characterize thousands of coding and non-coding variants in a single experiment with high reproducibility. To showcase its applications, we first identified essential nucleotides for a 716 bp MYC enhancer via PRIME-mediated saturation mutagenesis. Next, we applied PRIME to functionally characterize 1,304 non-coding variants associated with breast cancer and 3,699 variants from ClinVar. We discovered that 103 non-coding variants and 156 variants of uncertain significance are functional via affecting cell fitness. Collectively, we demonstrate PRIME capable of characterizing genetic variants at base-pair resolution and scale, advancing accurate genome annotation for disease risk prediction, diagnosis, and therapeutic target identification.
]]></description>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Nierenberg, J. L.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Beaman, C.</dc:creator>
<dc:creator>Pham, T.</dc:creator>
<dc:creator>Nobuhara, M.</dc:creator>
<dc:creator>Takagi, M. A.</dc:creator>
<dc:creator>Narayan, V.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Ziv, E.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:date>2023-07-12</dc:date>
<dc:identifier>doi:10.1101/2023.07.12.548736</dc:identifier>
<dc:title><![CDATA[High throughput PRIME editing screens identify functional DNA variants in the human genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.12.548753v1?rss=1">
<title>
<![CDATA[
The h-current controls cortical recurrent network activity through modulation of dendrosomatic communication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.12.548753v1?rss=1</link>
<description><![CDATA[
A fundamental feature of the cerebral cortex is the ability to rapidly turn on and off maintained activity within ensembles of neurons through recurrent excitation balanced by inhibition. Here we demonstrate that reduction of the h-current, which is especially prominent in pyramidal cell dendrites, strongly increases the ability of local cortical networks to generate maintained recurrent activity. Reduction of the h-current resulted in hyperpolarization and increase in input resistance of both the somata and apical dendrites of layer 5 pyramidal cells, while strongly increasing the dendrosomatic transfer of low (<20 Hz) frequencies, causing an increased responsiveness to dynamic clamp-induced recurrent network-like activity injected into the dendrites and substantially increasing the duration of spontaneous Up states. We propose that modulation of the h-current may strongly control the ability of cortical networks to generate recurrent persistent activity and the formation and dissolution of neuronal ensembles.
]]></description>
<dc:creator>McCormick, D. A.</dc:creator>
<dc:creator>Shu, Y.</dc:creator>
<dc:creator>Hasenstaub, A. R.</dc:creator>
<dc:date>2023-07-12</dc:date>
<dc:identifier>doi:10.1101/2023.07.12.548753</dc:identifier>
<dc:title><![CDATA[The h-current controls cortical recurrent network activity through modulation of dendrosomatic communication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.13.548459v1?rss=1">
<title>
<![CDATA[
Radial glia control microglial differentiation via integrin avb8-dependent trans-activation of TGFB1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.13.548459v1?rss=1</link>
<description><![CDATA[
Microglia diversity emerges from interactions between intrinsic genetic programs and environment-derived signals, but how these processes unfold and interact in the developing brain remains unclear. Here, we show that radial glia-expressed integrin beta 8 (ITGB8) expressed in radial glia progenitors activates microglia-expressed TGF{beta}1, permitting microglial development. Domain-restricted deletion of Itgb8 in these progenitors establishes complementary regions with developmentally arrested "dysmature" microglia that persist into adulthood. In the absence of autocrine TGF{beta}1 signaling, we find that microglia adopt a similar dysmature phenotype, leading to neuromotor symptoms almost identical to Itgb8 mutant mice. In contrast, microglia lacking the TGF{beta} signal transducers Smad2 and Smad3 have a less polarized dysmature phenotype and correspondingly less severe neuromotor dysfunction. Finally, we show that non-canonical (Smad-independent) signaling partially suppresses disease and development associated gene expression, providing compelling evidence for the adoption of microglial developmental signaling pathways in the context of injury or disease.
]]></description>
<dc:creator>McKinsey, G. L.</dc:creator>
<dc:creator>Santander, N.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Kleemann, K.</dc:creator>
<dc:creator>Katewa, A.</dc:creator>
<dc:creator>Tran, L.</dc:creator>
<dc:creator>Conant, K.</dc:creator>
<dc:creator>Barraza, M.</dc:creator>
<dc:creator>Lizama, C. O.</dc:creator>
<dc:creator>Koo, H. J.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Mukherjee, D.</dc:creator>
<dc:creator>Paidassi, H.</dc:creator>
<dc:creator>Anton, E. S.</dc:creator>
<dc:creator>Atabai, K.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:creator>Butovsky, O.</dc:creator>
<dc:creator>Arnold, T. D.</dc:creator>
<dc:date>2023-07-13</dc:date>
<dc:identifier>doi:10.1101/2023.07.13.548459</dc:identifier>
<dc:title><![CDATA[Radial glia control microglial differentiation via integrin avb8-dependent trans-activation of TGFB1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.12.548702v1?rss=1">
<title>
<![CDATA[
Validation of human telomere length trans-ancestry meta-analysis association signals identifies POP5 and KBTBD6 as novel human telomere length regulation genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.12.548702v1?rss=1</link>
<description><![CDATA[
Telomere length genome-wide association studies (GWAS) have become well-powered to detect novel genes in telomere length regulation. However, no prior work has validated these putative novel genes to confirm the contribution of GWAS loci to telomere length regulation. We conducted a trans-ancestry meta-analysis of 211,369 individuals. Through enrichment analyses of chromatin state and cell-type heritability we identified blood and immune cells as the most relevant cell type to examine telomere length association signals. We validated specific GWAS associations by overexpressing KBTBD6, a component of an E3 ubiquitin ligase complex, and POP5, a component of the Ribonuclease P/MRP complex, and demonstrating that both lengthened telomeres as predicted by our statistical analyses. CRISPR/Cas9 deletion of the predicted causal regions of these association peaks in K562 immortalized blood cells reduced expression of these genes, demonstrating that these loci are related to transcriptional regulation of KBTBD6 and POP5, respectively. Together our results demonstrate the utility of telomere length GWAS in the identification of novel telomere length regulation mechanisms and highlight the importance of the proteasome-ubiquitin pathway in telomere length regulation.
]]></description>
<dc:creator>Keener, R.</dc:creator>
<dc:creator>Chhetri, S. B.</dc:creator>
<dc:creator>Connelly, C. J.</dc:creator>
<dc:creator>Taub, M. A.</dc:creator>
<dc:creator>Conomos, M. P.</dc:creator>
<dc:creator>Weinstock, J. S.</dc:creator>
<dc:creator>Ni, B.</dc:creator>
<dc:creator>Strober, B. J.</dc:creator>
<dc:creator>Aslibekyan, S.</dc:creator>
<dc:creator>Auer, P. L.</dc:creator>
<dc:creator>Barwick, L.</dc:creator>
<dc:creator>Becker, L. C.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Bleecker, E. R.</dc:creator>
<dc:creator>Brody, J. A.</dc:creator>
<dc:creator>Cade, B. E.</dc:creator>
<dc:creator>Celedon, J. C.</dc:creator>
<dc:creator>Chang, Y.-C.</dc:creator>
<dc:creator>Cupples, L. A.</dc:creator>
<dc:creator>Custer, B.</dc:creator>
<dc:creator>Freedman, B. I.</dc:creator>
<dc:creator>Gladwin, M. T.</dc:creator>
<dc:creator>Heckbert, S. R.</dc:creator>
<dc:creator>Hou, L.</dc:creator>
<dc:creator>Irvin, M. R.</dc:creator>
<dc:creator>Isasi, C. R.</dc:creator>
<dc:creator>Johnsen, J. M.</dc:creator>
<dc:creator>Kenny, E. E.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Minster, R. L.</dc:creator>
<dc:creator>Nekhai, S.</dc:creator>
<dc:creator>Pankratz, N.</dc:creator>
<dc:creator>Peyser, P. A.</dc:creator>
<dc:creator>Rotter, J. I.</dc:creator>
<dc:creator>Taylor, K. D.</dc:creator>
<dc:creator>Telen, M. J.</dc:creator>
<dc:creator>Wu, B.</dc:creator>
<dc:creator>Yanek, L. R.</dc:creator>
<dc:creator>Yang, I. V.</dc:creator>
<dc:creator>Albert, C.</dc:creator>
<dc:creator>Arnett, D. K.</dc:creator>
<dc:creator>Ashley-Koch, A.</dc:creator>
<dc:date>2023-07-14</dc:date>
<dc:identifier>doi:10.1101/2023.07.12.548702</dc:identifier>
<dc:title><![CDATA[Validation of human telomere length trans-ancestry meta-analysis association signals identifies POP5 and KBTBD6 as novel human telomere length regulation genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.13.548942v1?rss=1">
<title>
<![CDATA[
Monitoring integrated stress response in live Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.13.548942v1?rss=1</link>
<description><![CDATA[
Cells exhibit stress responses to various environmental changes. Among these responses, the integrated stress response (ISR) plays a pivotal role as a crucial stress signaling pathway. While extensive ISR research has been conducted on cultured cells, our understanding of its implications in multicellular organisms remains limited, largely due to the constraints of current techniques that hinder our ability to track and manipulate the ISR in vivo. To overcome these limitations, we have successfully developed an internal ribosome entry site (IRES)-based fluorescent reporter system. This innovative reporter enables us to label Drosophila cells, within the context of a living organism, that exhibit eIF2 phosphorylation-dependent translational shutoff - a characteristic feature of the ISR and viral infections. Through this methodology, we have unveiled tissue- and cell-specific regulation of stress response in Drosophila flies and have even been able to detect stressed tissues in vivo during virus and bacterial infections. To further validate the specificity of our reporter, we have engineered ISR-null eIF2S50A mutant flies for stress response analysis. Our results shed light on the tremendous potential of this technique for investigating a broad range of developmental, stress, and infection-related experimental conditions. Combining the reporter tool with ISR-null mutants establishes Drosophila as an exceptionally powerful model for studying the ISR in the context of multicellular organisms.
]]></description>
<dc:creator>Lidsky, P. V.</dc:creator>
<dc:creator>Yuan, J.</dc:creator>
<dc:creator>Lashkevich, K. A.</dc:creator>
<dc:creator>Dmitriev, S. E.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:date>2023-07-14</dc:date>
<dc:identifier>doi:10.1101/2023.07.13.548942</dc:identifier>
<dc:title><![CDATA[Monitoring integrated stress response in live Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.14.549074v1?rss=1">
<title>
<![CDATA[
Tissue tension permits β-catenin phosphorylation to drive mesoderm specification in human embryonic stem cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.14.549074v1?rss=1</link>
<description><![CDATA[
The role of morphogenetic forces in cell fate specification is an area of intense interest. Our prior studies suggested that the development of high cell-cell tension in human embryonic stem cells (hESC) colonies permits the Src-mediated phosphorylation of junctional {beta}-catenin that accelerates its release to potentiate Wnt-dependent signaling critical for initiating mesoderm specification. Using an ectopically expressed nonphosphorylatable mutant of {beta}-catenin (Y654F), we now provide direct evidence that impeding tension-dependent Src-mediated {beta}-catenin phosphorylation impedes the expression of Brachyury (T) and the epithelial-to-mesenchymal transition (EMT) necessary for mesoderm specification. Addition of exogenous Wnt3a or inhibiting GSK3{beta} activity rescued mesoderm expression, emphasizing the importance of force dependent Wnt signaling in regulating mechanomorphogenesis. Our work provides a framework for understanding tension-dependent {beta}-catenin/Wnt signaling in the self-organization of tissues during developmental processes including gastrulation.
]]></description>
<dc:creator>Ayad, N. M. E.</dc:creator>
<dc:creator>Lakins, J. N.</dc:creator>
<dc:creator>Ghagre, A.</dc:creator>
<dc:creator>Ehrlicher, A. J.</dc:creator>
<dc:creator>Weaver, V. M.</dc:creator>
<dc:date>2023-07-15</dc:date>
<dc:identifier>doi:10.1101/2023.07.14.549074</dc:identifier>
<dc:title><![CDATA[Tissue tension permits β-catenin phosphorylation to drive mesoderm specification in human embryonic stem cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.14.548818v1?rss=1">
<title>
<![CDATA[
Distinct senescence mechanisms restrain progression of dysplastic nevi 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.14.548818v1?rss=1</link>
<description><![CDATA[
TERT promoter mutations (TPMs) are frequently found in different cancer types, including approximately 70% of sun-exposed skin melanomas. In melanoma, TPMs are among the earliest mutations and can be present during the transition from nevus to melanoma. However, the specific factors that contribute to the selection of TPMs in certain nevi subsets are not well understood. To investigate this, we analyzed a group of dysplastic nevi (DN) by sequencing genes commonly mutated in melanocytic neoplasms. We examined the relationship between the identified mutations, patient age, telomere length, histological features, and the expression of p16. Our findings reveal that TPMs are more prevalent in DN from older patients and are associated with shorter telomeres. Importantly, these TPMs were not found in nevi with BRAF V600E mutations. Conversely, DN with BRAF V600E mutations were observed in younger patients, had longer telomeres, and a higher proportion of p16-positive cells. This suggests that these nevi arrest growth independently of telomere shortening through a mechanism known as oncogene-induced senescence (OIS). These characteristics extend to melanoma sequencing data sets, where melanomas with BRAF V600E mutations were more likely to have CDKN2A inactivation, overriding OIS. In contrast, melanomas without BRAF V600E mutations showed a higher frequency of TPMs. Our data imply that TPMs are selected to bypass replicative senescence (RS) in cells that were not arrested by OIS. Overall, our results indicate that a subset of melanocytic neoplasms face constraints from RS, while others encounter OIS and RS. The order in which these barriers are overcome during progression to melanoma depends on the mutational context.
]]></description>
<dc:creator>Lorbeer, F. K.</dc:creator>
<dc:creator>Rieser, G.</dc:creator>
<dc:creator>Goel, A.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Oh, A.</dc:creator>
<dc:creator>Yeh, I.</dc:creator>
<dc:creator>Bastian, B. C.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:date>2023-07-16</dc:date>
<dc:identifier>doi:10.1101/2023.07.14.548818</dc:identifier>
<dc:title><![CDATA[Distinct senescence mechanisms restrain progression of dysplastic nevi]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.17.549397v1?rss=1">
<title>
<![CDATA[
Simultaneous multi-site editing of individual genomes using retron arrays 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.17.549397v1?rss=1</link>
<description><![CDATA[
Our understanding of genomics is limited by the scale of our genomic technologies. While libraries of genomic manipulations scaffolded on CRISPR gRNAs have been transformative, these existing approaches are typically multiplexed across genomes. Yet much of the complexity of real genomes is encoded within a genome across sites. Unfortunately, building cells with multiple, non-adjacent precise mutations remains a laborious cycle of editing, isolating an edited cell, and editing again. Here, we describe a technology for precisely modifying multiple sites on a single genome simultaneously. This technology - termed a multitron - is built from a heavily modified retron, in which multiple donor-encoding msds are produced from a single transcript. The multitron architecture is compatible with both recombineering in prokaryotic cells and CRISPR editing in eukaryotic cells. We demonstrate applications for this approach in molecular recording, genetic element minimization, and metabolic engineering.
]]></description>
<dc:creator>Gonzalez-Delgado, A.</dc:creator>
<dc:creator>Lopez, S. C.</dc:creator>
<dc:creator>Rojas-Montero, M.</dc:creator>
<dc:creator>Fishman, C. B.</dc:creator>
<dc:creator>Shipman, S. L.</dc:creator>
<dc:date>2023-07-17</dc:date>
<dc:identifier>doi:10.1101/2023.07.17.549397</dc:identifier>
<dc:title><![CDATA[Simultaneous multi-site editing of individual genomes using retron arrays]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.17.549407v1?rss=1">
<title>
<![CDATA[
Interaction with the membrane-anchored protein CHIC2 constrains the ubiquitin ligase activity of CHIP 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.17.549407v1?rss=1</link>
<description><![CDATA[
Maintenance of cellular health requires the proper regulation of E3 ubiquitin ligases. The E3 ligase CHIP is canonically regulated by its interactions with the molecular chaperones Hsp70 and Hsp90, which focus CHIPs ubiquitination activity on misfolded proteins. Here, we report a chaperone-independent interaction of CHIP with the membrane-anchored protein CHIC2, which strongly attenuates CHIPs ligase activity. We show that CHIC2 outcompetes abundant, cytosolic chaperones through its exquisite CHIP selectivity, rather than through enhanced affinity. In proteomic experiments, we find that CHIC2 knockout phenocopies CHIP knockout in certain cell types, implying that chaperone-independent interactions can sometimes predominate CHIPs functions. Furthermore, loss of the CHIP-CHIC2 interaction induces neurodegeneration and shortens lifespan in C. elegans, demonstrating that formation of this chaperone-independent complex is important in animals. We propose that CHIC2 attenuates CHIP activity at the membrane, offering a novel mechanism by which this ubiquitin ligase can be regulated.
]]></description>
<dc:creator>Callahan, M. D.</dc:creator>
<dc:creator>Hodul, M.</dc:creator>
<dc:creator>Carroll, E. C.</dc:creator>
<dc:creator>Ravalin, M.</dc:creator>
<dc:creator>Nadel, C. M.</dc:creator>
<dc:creator>de Silva, A. R.</dc:creator>
<dc:creator>Cupo, A. R.</dc:creator>
<dc:creator>McDermott, L. E.</dc:creator>
<dc:creator>Nix, J. C.</dc:creator>
<dc:creator>Page, R. C.</dc:creator>
<dc:creator>Kao, A. W.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:date>2023-07-17</dc:date>
<dc:identifier>doi:10.1101/2023.07.17.549407</dc:identifier>
<dc:title><![CDATA[Interaction with the membrane-anchored protein CHIC2 constrains the ubiquitin ligase activity of CHIP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.16.549192v1?rss=1">
<title>
<![CDATA[
Sexually dimorphic differences in angiogenesis markers predict brain aging trajectories 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.16.549192v1?rss=1</link>
<description><![CDATA[
Aberrant angiogenesis could contribute to cognitive impairment, representing a therapeutic target for preventing dementia. However, most angiogenesis studies focus on model organisms. To test the relevance of angiogenesis to human cognitive aging, we evaluated associations of circulating blood markers of angiogenesis with brain aging trajectories in two deeply phenotyped human cohorts (n=435, age 74+9) with longitudinal cognitive assessments, biospecimens, structural brain imaging, and clinical data. Machine learning and traditional statistics revealed sex dimorphic associations of plasma angiogenic growth factors with brain aging outcomes. Specifically, angiogenesis is associated with higher executive function and less brain atrophy in younger women (not men), a directionality of association that reverses around age 75. Higher levels of basic fibroblast growth factor, known for pleiotropic effects on multiple cell types, predicted favorable cognitive trajectories. This work demonstrates the relevance of angiogenesis to brain aging with important therapeutic implications for vascular cognitive impairment and dementia.
]]></description>
<dc:creator>Torres-Espin, A.</dc:creator>
<dc:creator>Rabadaugh, H.</dc:creator>
<dc:creator>Fitzsimons, S.</dc:creator>
<dc:creator>Chou, A.</dc:creator>
<dc:creator>Harvey, D.</dc:creator>
<dc:creator>Lindberg, C.</dc:creator>
<dc:creator>Goldberger, L.</dc:creator>
<dc:creator>Casaletto, K. B.</dc:creator>
<dc:creator>Staffaroni, A.</dc:creator>
<dc:creator>Maillard, P.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Hinman, J. D.</dc:creator>
<dc:creator>DeCarli, C.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Elahi, F. M.</dc:creator>
<dc:date>2023-07-18</dc:date>
<dc:identifier>doi:10.1101/2023.07.16.549192</dc:identifier>
<dc:title><![CDATA[Sexually dimorphic differences in angiogenesis markers predict brain aging trajectories]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.17.549377v1?rss=1">
<title>
<![CDATA[
C9orf72 gene networks in the human brain correlate with cortical thickness in C9-FTD and implicate vulnerable cell types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.17.549377v1?rss=1</link>
<description><![CDATA[
IntroductionA hexanucleotide repeat expansion (HRE) intronic to chromosome 9 open reading frame 72 (C9orf72) is recognized as the most common genetic cause of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), and ALS-FTD. Identifying genes that show similar regional co-expression patterns to C9orf72 may help identify novel gene targets and biological mechanisms that mediate selective vulnerability to ALS and FTD pathogenesis.

MethodsWe leveraged mRNA expression data in healthy brain from the Allen Human Brain Atlas to evaluate C9orf72 co-expression patterns. To do this, we correlated average C9orf72 expression values in 51 regions across different anatomical divisions (cortex, subcortex, cerebellum) with average gene expression values for 15,633 protein-coding genes, including 50 genes known to be associated with ALS, FTD, or ALS-FTD. We then evaluated whether the identified C9orf72 co-expressed genes correlated with patterns of cortical thickness in symptomatic C9orf72 pathogenic HRE carriers (n=19). Lastly, we explored whether genes with significant C9orf72 radiogenomic correlations (i.e.,  C9orf72 gene network) were enriched in specific cell populations in the brain and enriched for specific biological and molecular pathways.

ResultsA total of 1,748 genes showed an anatomical distribution of gene expression in the brain similar to C9orf72 and significantly correlated with patterns of cortical thickness in C9orf72 HRE carriers. This C9orf72 gene network was differentially expressed in cell populations previously implicated in ALS and FTD, including layer 5b cells, cholinergic motor neurons in the spinal cord, and medium spiny neurons of the striatum, and was enriched for biological and molecular pathways associated with multiple neurotransmitter systems, protein ubiquitination, autophagy, and MAPK signaling, among others.

ConclusionsConsidered together, we identified a network of C9orf72-associated genes that may influence selective regional and cell-type-specific vulnerabilities in ALS/FTD.
]]></description>
<dc:creator>Broce, I.</dc:creator>
<dc:creator>Sirkis, D.</dc:creator>
<dc:creator>Nillo, R. M.</dc:creator>
<dc:creator>Bonham, L. W.</dc:creator>
<dc:creator>Lee, S. E.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Sturm, V.</dc:creator>
<dc:creator>Sugrue, L. S.</dc:creator>
<dc:creator>Desikan, R.</dc:creator>
<dc:creator>Yokoyama, J. S.</dc:creator>
<dc:date>2023-07-19</dc:date>
<dc:identifier>doi:10.1101/2023.07.17.549377</dc:identifier>
<dc:title><![CDATA[C9orf72 gene networks in the human brain correlate with cortical thickness in C9-FTD and implicate vulnerable cell types]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.18.549551v1?rss=1">
<title>
<![CDATA[
High-throughput complement component 4 genomic sequence analysis with C4Investigator 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.18.549551v1?rss=1</link>
<description><![CDATA[
The complement component 4 gene locus, composed of the C4A and C4B genes and located on chromosome 6, encodes for C4 protein, a key intermediate in the classical and lectin pathways of the complement system. The complement system is an important modulator of immune system activity and is also involved in the clearance of immune complexes and cellular debris. The C4 gene locus exhibits copy number variation, with each composite gene varying between 0-5 copies per haplotype, C4 genes also vary in size depending on the presence of the HERV retrovirus in intron 9, denoted by C4(L) for long-form and C4(S) for short-form, which modulates expression and is found in both C4A and C4B. Additionally, human blood group antigens Rodgers and Chido are located on the C4 protein, with the Rodger epitope generally found on C4A protein, and the Chido epitope generally found on C4B protein. C4 copy number variation has been implicated in numerous autoimmune and pathogenic diseases. Despite the central role of C4 in immune function and regulation, high-throughput genomic sequence analysis of C4 variants has been impeded by the high degree of sequence similarity and complex genetic variation exhibited by these genes. To investigate C4 variation using genomic sequencing data, we have developed a novel bioinformatic pipeline for comprehensive, high-throughput characterization of human C4 sequence from short-read sequencing data, named C4Investigator. Using paired-end targeted or whole genome sequence data as input, C4Investigator determines gene copy number for overall C4, C4A, C4B, C4(Rodger), C4(Ch), C4(L), and C4(S), additionally, C4Ivestigator reports the full overall C4 aligned sequence, enabling nucleotide level analysis of C4. To demonstrate the utility of this workflow we have analyzed C4 variation in the 1000 Genomes Project Dataset, showing that the C4 genes are highly poly-allelic with many variants that have the potential to impact C4 protein function.
]]></description>
<dc:creator>Marin, W. M.</dc:creator>
<dc:creator>Augusto, D. G.</dc:creator>
<dc:creator>Wade, K. J.</dc:creator>
<dc:creator>Hollenbach, J. A.</dc:creator>
<dc:date>2023-07-19</dc:date>
<dc:identifier>doi:10.1101/2023.07.18.549551</dc:identifier>
<dc:title><![CDATA[High-throughput complement component 4 genomic sequence analysis with C4Investigator]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.19.549443v1?rss=1">
<title>
<![CDATA[
Genome editing induces the transfer of mitochondrial DNA into the nuclear genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.19.549443v1?rss=1</link>
<description><![CDATA[
Mitochondria serve as the cellular powerhouse, and their distinct DNA makes them a prospective target for gene editing to treat genetic disorders. However, the impact of genome editing on mitochondrial DNA (mtDNA) stability remains a mystery. Our study reveals previously unknown risks of genome editing that both nuclear and mitochondrial editing cause broad transfer of mitochondrial DNA segments into the nuclear genome in various cell types including human cell lines, primary T cells, retinal cells, and mouse embryos. Furthermore, drug-induced mitochondrial stresses and mtDNA breaks exacerbate this transfer of mtDNA into the nuclear genome. Notably, we observe that the newly developed mitochondrial base editor DdCBE can also cause widespread mtDNA integrations. However, we provide a practical solution to suppress the transfer of mtDNA by co-expressing TREX1 or TREX2 exonucleases during DdCBE editing. These findings also shed light on the origins of mitochondrial-nuclear DNA segments.
]]></description>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Ou, L.</dc:creator>
<dc:creator>Gan, T.</dc:creator>
<dc:creator>Zhangding, Z.</dc:creator>
<dc:creator>Yuan, S.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Yin, J.</dc:creator>
<dc:creator>Xin, C.</dc:creator>
<dc:creator>Tian, Y.</dc:creator>
<dc:creator>Hu, J.</dc:creator>
<dc:date>2023-07-20</dc:date>
<dc:identifier>doi:10.1101/2023.07.19.549443</dc:identifier>
<dc:title><![CDATA[Genome editing induces the transfer of mitochondrial DNA into the nuclear genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.20.549807v1?rss=1">
<title>
<![CDATA[
Genetic complexity alters drug susceptibility of asexual and gametocyte stages of Plasmodium falciparum to antimalarial candidates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.20.549807v1?rss=1</link>
<description><![CDATA[
Withdrawal statementThe authors have withdrawn their manuscript owing to internal reviewing and agreement to submission by all parties involved in the work not yet completed on this version of the paper. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.
]]></description>
<dc:creator>Greyling, N.</dc:creator>
<dc:creator>van der Watt, M.</dc:creator>
<dc:creator>Gwarinda, H.</dc:creator>
<dc:creator>van Heerden, A.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:creator>Leroy, D.</dc:creator>
<dc:creator>Niemand, J.</dc:creator>
<dc:creator>Birkholtz, L.-M.</dc:creator>
<dc:date>2023-07-21</dc:date>
<dc:identifier>doi:10.1101/2023.07.20.549807</dc:identifier>
<dc:title><![CDATA[Genetic complexity alters drug susceptibility of asexual and gametocyte stages of Plasmodium falciparum to antimalarial candidates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.21.550046v1?rss=1">
<title>
<![CDATA[
Structure of a tripartite protein complex that targets toxins to the type VII secretion system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.21.550046v1?rss=1</link>
<description><![CDATA[
Type VII secretion systems are membrane-embedded nanomachines used by Gram-positive bacteria to export effector proteins from the cytoplasm to the extracellular environment. Many of these effectors are polymorphic toxins comprised of an N-terminal Leu-x-Gly (LXG) domain of unknown function and a C-terminal toxin domain that inhibits the growth of bacterial competitors. In recent work, it was shown that LXG effectors require two cognate Lap proteins for T7SS-dependent export. Here, we present the 2.6[A] structure of the LXG domain of the TelA toxin from the opportunistic pathogen Streptococcus intermedius in complex with both of its cognate Lap targeting factors. The structure reveals an elongated -helical bundle within which each Lap protein makes extensive hydrophobic contacts with either end of the LXG domain. Remarkably, despite low overall sequence identity, we identify striking structural similarity between our LXG complex and PE-PPE heterodimers exported by the distantly related ESX type VII secretion systems of Mycobacteria implying a conserved mechanism of effector export among diverse Gram-positive bacteria. Overall, our findings demonstrate that LXG domains, in conjunction with their cognate Lap targeting factors, represent a tripartite secretion signal for a widespread family of T7SS toxins.
]]></description>
<dc:creator>Klein, T. A.</dc:creator>
<dc:creator>Shah, P. Y.</dc:creator>
<dc:creator>Gkragkopoulou, P.</dc:creator>
<dc:creator>Grebenc, D. W.</dc:creator>
<dc:creator>Kim, Y.</dc:creator>
<dc:creator>Whitney, J. C.</dc:creator>
<dc:date>2023-07-21</dc:date>
<dc:identifier>doi:10.1101/2023.07.21.550046</dc:identifier>
<dc:title><![CDATA[Structure of a tripartite protein complex that targets toxins to the type VII secretion system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.21.549930v1?rss=1">
<title>
<![CDATA[
APOE4/4 is linked to damaging lipid droplets in Alzheimer's microglia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.21.549930v1?rss=1</link>
<description><![CDATA[
Several genetic risk factors for Alzheimers Disease (AD) implicate genes involved in lipid metabolism and many of these lipid genes are highly expressed in glial cells. However, the relationship between lipid metabolism in glia and AD pathology remains poorly understood. Through single-nucleus RNA-sequencing of AD brain tissue, we have identified a microglial state defined by the expression of the lipid droplet (LD) associated enzyme ACSL1 with ACSL1-positive microglia most abundant in AD patients with the APOE4/4 genotype. In human iPSC-derived microglia (iMG) fibrillar A{beta} (fA{beta}) induces ACSL1 expression, triglyceride synthesis, and LD accumulation in an APOE-dependent manner. Additionally, conditioned media from LD-containing microglia leads to Tau phosphorylation and neurotoxicity in an APOE-dependent manner. Our findings suggest a link between genetic risk factors for AD with microglial LD accumulation and neurotoxic microglial-derived factors, potentially providing novel therapeutic strategies for AD.
]]></description>
<dc:creator>Haney, M.</dc:creator>
<dc:creator>Palovics, R.</dc:creator>
<dc:creator>Munson, C.</dc:creator>
<dc:creator>Long, C.</dc:creator>
<dc:creator>Johansson, P.</dc:creator>
<dc:creator>Yip, O.</dc:creator>
<dc:creator>Dong, W.</dc:creator>
<dc:creator>Rawat, E.</dc:creator>
<dc:creator>West, E.</dc:creator>
<dc:creator>Schlachetzki, J.</dc:creator>
<dc:creator>Tsai, A. P.-Y.</dc:creator>
<dc:creator>Guldner, I.</dc:creator>
<dc:creator>Lamichhane, B.</dc:creator>
<dc:creator>Smith, A.</dc:creator>
<dc:creator>Schaum, N.</dc:creator>
<dc:creator>Calcuttawala, K.</dc:creator>
<dc:creator>Shin, A.</dc:creator>
<dc:creator>Wang, Y.-H.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Koutsodendris, N.</dc:creator>
<dc:creator>Serrano, G. E.</dc:creator>
<dc:creator>Beach, T.</dc:creator>
<dc:creator>Reiman, E. M.</dc:creator>
<dc:creator>Glass, C.</dc:creator>
<dc:creator>Abu-Remaileh, M. M.</dc:creator>
<dc:creator>Enejder, A.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Wyss-Coray, T.</dc:creator>
<dc:date>2023-07-25</dc:date>
<dc:identifier>doi:10.1101/2023.07.21.549930</dc:identifier>
<dc:title><![CDATA[APOE4/4 is linked to damaging lipid droplets in Alzheimer's microglia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.25.550452v1?rss=1">
<title>
<![CDATA[
Early-life stress triggers long-lasting organismal resilience and longevity via tetraspanin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.25.550452v1?rss=1</link>
<description><![CDATA[
Early-life stress experiences can produce lasting impacts on organismal adaptation and fitness. How transient stress elicits memory-like physiological effects is largely unknown. Here we show that early-life thermal stress strongly up-regulates tsp-1, a gene encoding the conserved transmembrane tetraspanin in C. elegans. TSP-1 forms prominent multimers and stable web- like structures critical for membrane barrier functions in adults and during aging. The up- regulation of TSP-1 persists even after transient early-life stress. Such regulation requires CBP- 1, a histone acetyl-transferase that facilitates initial tsp-1 transcription. Tetraspanin webs form regular membrane structures and mediate resilience-promoting effects of early-life thermal stress. Gain-of-function TSP-1 confers marked C. elegans longevity extension and thermal resilience in human cells. Together, our results reveal a cellular mechanism by which early-life thermal stress produces long-lasting memory-like impact on organismal resilience and longevity.

TeaserStudies reveal mechanisms of how early-life heat exposure produces long-lasting benefits on longevity in the nematode C. elegans.
]]></description>
<dc:creator>Jiang, W. I.</dc:creator>
<dc:creator>Belly, H. D.</dc:creator>
<dc:creator>Wang, B. K.</dc:creator>
<dc:creator>Wong, A.</dc:creator>
<dc:creator>Kim, M.</dc:creator>
<dc:creator>Oh, F.</dc:creator>
<dc:creator>DeGeorge, J.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Guang, S.</dc:creator>
<dc:creator>Weiner, O. K.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2023-07-25</dc:date>
<dc:identifier>doi:10.1101/2023.07.25.550452</dc:identifier>
<dc:title><![CDATA[Early-life stress triggers long-lasting organismal resilience and longevity via tetraspanin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.24.550427v1?rss=1">
<title>
<![CDATA[
Direct reprogramming of oral epithelial progenitor cells to cancer stem cells at single cell resolution in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.24.550427v1?rss=1</link>
<description><![CDATA[
Tumor initiation represents the first step in tumorigenesis during which normal progenitor cells undergo cell fate transition to cancer. Capturing this process as it occurs in vivo, however, remains elusive. Here we employ spatiotemporally controlled oncogene activation and tumor suppressor inhibition together with multiomics to unveil the processes underlying oral epithelial progenitor cell reprogramming into tumor initiating cells (TIC) at single cell resolution. TIC displayed a distinct stem-like state, defined by aberrant proliferative, hypoxic, squamous differentiation, and partial epithelial to mesenchymal (pEMT) invasive gene programs. YAP-mediated TIC programs included the activation of oncogenic transcriptional networks and mTOR signaling, and the recruitment of myeloid cells to the invasive front contributing to tumor infiltration. TIC transcriptional programs are conserved in human head and neck cancer and associated with poor patient survival. These findings illuminate processes underlying cancer initiation at single cell resolution, and identify candidate targets for early cancer detection and prevention.
]]></description>
<dc:creator>Faraji, F.</dc:creator>
<dc:creator>Ramirez, S. I.</dc:creator>
<dc:creator>Clubb, L.</dc:creator>
<dc:creator>Sato, K.</dc:creator>
<dc:creator>Anguiano Quiroz, P. Y.</dc:creator>
<dc:creator>Galloway, W. M.</dc:creator>
<dc:creator>Mikulski, Z.</dc:creator>
<dc:creator>Hoang, T. S.</dc:creator>
<dc:creator>Medetgul-Ernar, K.</dc:creator>
<dc:creator>Marangoni, P.</dc:creator>
<dc:creator>Jones, K. B.</dc:creator>
<dc:creator>Officer, A.</dc:creator>
<dc:creator>Molinolo, A. A.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Sakaguchi, K.</dc:creator>
<dc:creator>Califano, J. A.</dc:creator>
<dc:creator>Smith, Q.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Tamayo, P.</dc:creator>
<dc:creator>Gutkind, J. S.</dc:creator>
<dc:date>2023-07-26</dc:date>
<dc:identifier>doi:10.1101/2023.07.24.550427</dc:identifier>
<dc:title><![CDATA[Direct reprogramming of oral epithelial progenitor cells to cancer stem cells at single cell resolution in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.25.550557v1?rss=1">
<title>
<![CDATA[
Human Plasma Proteomic Profile of Clonal Hematopoiesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.25.550557v1?rss=1</link>
<description><![CDATA[
Plasma proteomic profiles associated with subclinical somatic mutations in blood cells may offer novel insights into downstream clinical consequences. Here, we explore such patterns in clonal hematopoiesis of indeterminate potential (CHIP), which is linked to several cancer and non-cancer outcomes, including coronary artery disease (CAD). Among 61,833 ancestrally diverse participants (3,881 with CHIP) from NHLBI TOPMed and UK Biobank with blood-based DNA sequencing and proteomic measurements (1,148 proteins by SomaScan in TOPMed and 2,917 proteins by Olink in UK Biobank), we identified 32 and 345 unique proteins from TOPMed and UK Biobank, respectively, associated with the most prevalent driver genes (DNMT3A, TET2, and ASXL1). These associations showed substantial heterogeneity by driver genes, sex, and race, and were enriched for immune response and inflammation pathways. Mendelian randomization in humans, coupled with ELISA in hematopoietic Tet2-/- vs wild-type mice validation, disentangled causal proteomic perturbations from TET2 CHIP. Lastly, we identified plasma proteins shared between CHIP and CAD.
]]></description>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Vromman, A.</dc:creator>
<dc:creator>Nguyen, N. Q. H.</dc:creator>
<dc:creator>Vellarikkal, S. K.</dc:creator>
<dc:creator>Uddin, M. M.</dc:creator>
<dc:creator>Niroula, A.</dc:creator>
<dc:creator>Griffin, G.</dc:creator>
<dc:creator>Honigberg, M. C.</dc:creator>
<dc:creator>Lin, A. E.</dc:creator>
<dc:creator>Gibson, C. J.</dc:creator>
<dc:creator>Katz, D. H.</dc:creator>
<dc:creator>Tahir, U.</dc:creator>
<dc:creator>Haidermota, S.</dc:creator>
<dc:creator>Ganesh, S.</dc:creator>
<dc:creator>Antoine, T.</dc:creator>
<dc:creator>Weinstock, J.</dc:creator>
<dc:creator>Austin, T. R.</dc:creator>
<dc:creator>Vasan, R. S.</dc:creator>
<dc:creator>peloso, G. M.</dc:creator>
<dc:creator>Hornsby, W.</dc:creator>
<dc:creator>Ganz, P.</dc:creator>
<dc:creator>Manson, J. E.</dc:creator>
<dc:creator>Haring, B.</dc:creator>
<dc:creator>Kooperberg, C. L.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Psaty, B. M.</dc:creator>
<dc:creator>Min, Y.-I.</dc:creator>
<dc:creator>Correa, A.</dc:creator>
<dc:creator>Lange, L. A.</dc:creator>
<dc:creator>Post, W.</dc:creator>
<dc:creator>Rotter, J. I.</dc:creator>
<dc:creator>Rich, S. S.</dc:creator>
<dc:creator>Wilson, J. G.</dc:creator>
<dc:creator>Ebert, B. L.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Ballantyne, C. M.</dc:creator>
<dc:creator>Coresh, J.</dc:creator>
<dc:creator>Sankaran, V. G.</dc:creator>
<dc:creator>Bick, A. G.</dc:creator>
<dc:creator>Jaiswal, S.</dc:creator>
<dc:creator>Libby, P.</dc:creator>
<dc:creator>Gerszten, R. E.</dc:creator>
<dc:creator>NHLBI T</dc:creator>
<dc:date>2023-07-27</dc:date>
<dc:identifier>doi:10.1101/2023.07.25.550557</dc:identifier>
<dc:title><![CDATA[Human Plasma Proteomic Profile of Clonal Hematopoiesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.27.550738v1?rss=1">
<title>
<![CDATA[
Site-specific proximity labeling at single residue resolution for identification of protein partners in vitro and on cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.27.550738v1?rss=1</link>
<description><![CDATA[
The cell surface proteome, or surfaceome, is encoded by more than 4000 genes, but we are only beginning to understand the complexes they form. Rapid proximity labeling around specific membrane targets allows for capturing weak and transient interactions expected in the crowded and dynamic environment of the surfaceome. Recently, a high-resolution approach called Map has been described (Geri, J. B., Oakley, J. V., Reyes-Robles, T., Wang, T., McCarver, S. J., White, C. H., Rodriguez-Rivera, F. P., Parker, D. L., Hett, E. C., Fadeyi, O. O., Oslund, R. C., and MacMillan, D. W. C. (2020) Science 367, 1091-1097) in which an iridium (Ir)-based photocatalyst is attached to a specific antibody to target labeling of neighbors utilizing light-activated generation of carbenes from diazirine compounds via Dexter Energy Transfer (DET). Here we studied and optimized the spatial resolution for the method using an oncoprotein complex between the antibody drug, trastuzumab (Traz), and its target HER2. A set of eight single site-specific Ir-catalytic centers were engineered into Traz to study intra- and inter-molecular labeling in vitro and on cells by mass spectrometry. From this structurally well-characterized complex we observed a maximum distance of [~]110 [A] for labeling. Labeling occurred almost uniformly over the full range of amino acids, unlike the residue specific labeling of other techniques. To examine on cell labeling that is specific to HER2 as opposed to simply being on the membrane, we compared the labeling patterns for the eight Traz-catalyst species to random labeling of membrane proteins using a metabolically integrated fatty acid catalyst. Our results identified 20 high confidence HER2 neighbors, many novel, that were more than 6-fold enriched compared to the non-specific membrane tethered catalyst. These studies define distance labeling parameters from single-site catalysts placed directly on the membrane target of interest, and more accurately compare to non-specific labeling to identify membrane complexes with higher confidence.
]]></description>
<dc:creator>Bartholow, T. G.</dc:creator>
<dc:creator>Burroughs, P.</dc:creator>
<dc:creator>Elledge, S. K.</dc:creator>
<dc:creator>Byrnes, J. R.</dc:creator>
<dc:creator>Kirkemo, L. L.</dc:creator>
<dc:creator>Garda, V.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2023-07-27</dc:date>
<dc:identifier>doi:10.1101/2023.07.27.550738</dc:identifier>
<dc:title><![CDATA[Site-specific proximity labeling at single residue resolution for identification of protein partners in vitro and on cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.26.550648v1?rss=1">
<title>
<![CDATA[
Ribosome subunit attrition and activation of the p53-MDM4 axis dominate the response of MLL-rearranged cancer cells to WDR5 WIN site inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.26.550648v1?rss=1</link>
<description><![CDATA[
The chromatin-associated protein WD Repeat Domain 5 (WDR5) is a promising target for cancer drug discovery, with most efforts blocking an arginine-binding cavity on the protein called the "WIN" site that tethers WDR5 to chromatin. WIN site inhibitors (WINi) are active against multiple cancer cell types in vitro, the most notable of which are those derived from MLL-rearranged (MLLr) leukemias. Peptidomimetic WINi were originally proposed to inhibit MLLr cells via dysregulation of genes connected to hematopoietic stem cell expansion. Our discovery and interrogation of small molecule WIN site inhibitors, however, revealed that they act in MLLr cell lines to suppress ribosome protein gene (RPG) transcription, induce nucleolar stress, and activate p53. Because there is no precedent for an anti-cancer strategy that specifically targets RPG expression, we took an integrated multi-omics approach to further interrogate the mechanism of action of WINi in MLLr cancer cells. We show that WINi induce depletion of the stock of ribosomes, accompanied by a broad yet modest translational choke and changes in alternative mRNA splicing that inactivate the p53 antagonist MDM4. We also show that WINi are synergistic with agents including venetoclax and BET-bromodomain inhibitors. Together, these studies reinforce the concept that WINi are a novel type of ribosome-directed anti-cancer therapy and provide a resource to support their clinical implementation in MLLr leukemias and other malignancies.
]]></description>
<dc:creator>Howard, G. C.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Rose, K. L.</dc:creator>
<dc:creator>Patel, P.</dc:creator>
<dc:creator>Tsui, T.</dc:creator>
<dc:creator>Florian, A. C.</dc:creator>
<dc:creator>Lorey, S. L.</dc:creator>
<dc:creator>Grieb, B. C.</dc:creator>
<dc:creator>Smith, B. N.</dc:creator>
<dc:creator>Slota, M. J.</dc:creator>
<dc:creator>Reynolds, E. M.</dc:creator>
<dc:creator>Goswami, S.</dc:creator>
<dc:creator>Savona, M. R.</dc:creator>
<dc:creator>Lee, T.</dc:creator>
<dc:creator>Fesik, S. W.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Tansey, W. P.</dc:creator>
<dc:date>2023-07-29</dc:date>
<dc:identifier>doi:10.1101/2023.07.26.550648</dc:identifier>
<dc:title><![CDATA[Ribosome subunit attrition and activation of the p53-MDM4 axis dominate the response of MLL-rearranged cancer cells to WDR5 WIN site inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.28.551038v1?rss=1">
<title>
<![CDATA[
Distinct human stem cell subpopulations drive adipogenesis and fibrosis in musculoskeletal injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.28.551038v1?rss=1</link>
<description><![CDATA[
Fibroadipogenic progenitors (FAPs) maintain healthy skeletal muscle in homeostasis but drive muscle degeneration in chronic injuries by promoting adipogenesis and fibrosis. To uncover how these stem cells switch from a pro-regenerative to pro-degenerative role we perform single-cell mRNA sequencing of human FAPs from healthy and injured human muscles across a spectrum of injury, focusing on rotator cuff tears. We identify multiple subpopulations with progenitor, adipogenic, or fibrogenic gene signatures. We utilize full spectrum flow cytometry to identify distinct FAP subpopulations based on highly multiplexed protein expression. Injury severity increases adipogenic commitment of FAP subpopulations and is driven by the downregulation of DLK1. Treatment of FAPs both in vitro and in vivo with DLK1 reduces adipogenesis and fatty infiltration, suggesting that during injury, reduced DLK1 within a subpopulation of FAPs may drive degeneration. This work highlights how stem cells perform varied functions depending on tissue context, by dynamically regulating subpopulation fate commitment, which can be targeted improve patient outcomes after injury.
]]></description>
<dc:creator>Garcia, S. M.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:creator>Lau, J.</dc:creator>
<dc:creator>Chi, H. M.</dc:creator>
<dc:creator>Lizarraga, M.</dc:creator>
<dc:creator>Davies, M. R.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Feeley, B. T.</dc:creator>
<dc:date>2023-07-29</dc:date>
<dc:identifier>doi:10.1101/2023.07.28.551038</dc:identifier>
<dc:title><![CDATA[Distinct human stem cell subpopulations drive adipogenesis and fibrosis in musculoskeletal injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.27.550388v1?rss=1">
<title>
<![CDATA[
Computational prediction of protein interactions on single cells by proximity sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.27.550388v1?rss=1</link>
<description><![CDATA[
Proximity sequencing (Prox-seq) measures gene expression, protein expression, and protein complexes at the single cell level, using information from dual-antibody binding events and a single cell sequencing readout. Prox-seq provides multi-dimensional phenotyping of single cells and was recently used to track the formation of receptor complexes during inflammatory signaling in macrophages and to discover a new interaction between CD9/CD8 proteins on naive T cells. The distribution of protein abundance affects identification of protein complexes in a complicated manner in dual-binding assays like Prox-seq. These effects are difficult to explore with experiments, yet important for accurate quantification of protein complexes. Here, we introduce a physical model for protein dimer formation on single cells and computationally evaluate several different methods for reducing background noise when quantifying protein complexes. Furthermore, we developed an improved method for analysis of Prox-seq single-cell data, which resulted in more accurate and robust quantification of protein complexes. Finally, our model offers a simple way to investigate the behavior of Prox-seq under various biological conditions and guide users toward selecting the best analysis method for their data.
]]></description>
<dc:creator>Xia, J.</dc:creator>
<dc:creator>Phan, H. V.</dc:creator>
<dc:creator>Vistain, L.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Khan, A. A.</dc:creator>
<dc:creator>Tay, S.</dc:creator>
<dc:date>2023-07-30</dc:date>
<dc:identifier>doi:10.1101/2023.07.27.550388</dc:identifier>
<dc:title><![CDATA[Computational prediction of protein interactions on single cells by proximity sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.29.551032v1?rss=1">
<title>
<![CDATA[
Conserved long noncoding RNA TILAM promotes liver fibrosis through interaction with PML in hepatic stellate cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.29.551032v1?rss=1</link>
<description><![CDATA[
Background & AimsFibrosis is the common endpoint for all forms of chronic liver injury, and progression of fibrosis leads to the development of end-stage liver disease. Activation of hepatic stellate cells (HSCs) and their transdifferentiation to myofibroblasts results in the accumulation of extracellular matrix (ECM) proteins that form the fibrotic scar. Long noncoding (lnc) RNAs regulate the activity of HSCs and may provide targets for fibrotic therapies.

MethodsWe identified lncRNA TILAM as expressed near COL1A1 in human HSCs and performed loss-of-function studies in human HSCs and liver organoids. Transcriptomic analyses of HSCs isolated from mice defined the murine ortholog of TILAM. We then generated Tilam-deficient GFP reporter mice and quantified fibrotic responses to carbon tetrachloride (CCl4) and choline-deficient L-amino acid defined high fat diet (CDA-HFD). Co-precipitation studies, mass spectrometry, and gene expression analyses identified protein partners of TILAM.

ResultsTILAM is conserved between human and mouse HSCs and regulates expression of ECM proteins, including collagen. Tilam is selectively induced in HSCs during the development of fibrosis in vivo. In both male and female mice, loss of Tilam results in reduced fibrosis in the setting of CCl4 and CDA-HFD injury models. TILAM interacts with promyelocytic leukemia protein (PML) to stabilize PML protein levels and promote the fibrotic activity of HSCs.

ConclusionTILAM is activated in HSCs and interacts with PML to drive the development of liver fibrosis. Depletion of TILAM may serve as a therapeutic approach to combat the development of end stage liver disease.
]]></description>
<dc:creator>Sun, C.</dc:creator>
<dc:creator>Zhou, C.</dc:creator>
<dc:creator>Daneshvar, K.</dc:creator>
<dc:creator>Kratkiewicz, A. J.</dc:creator>
<dc:creator>Ben Saad, A.</dc:creator>
<dc:creator>Hess, A.</dc:creator>
<dc:creator>Chen, J. Y.</dc:creator>
<dc:creator>Pondick, J. V.</dc:creator>
<dc:creator>York, S. R.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Moran, S. P.</dc:creator>
<dc:creator>Gentile, S.</dc:creator>
<dc:creator>Ur Rahman, R.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Sparks, R.</dc:creator>
<dc:creator>Habboub, T.</dc:creator>
<dc:creator>Kim, B.-M.</dc:creator>
<dc:creator>Choi, M.</dc:creator>
<dc:creator>Affo, S.</dc:creator>
<dc:creator>Schwabe, R. F.</dc:creator>
<dc:creator>Popov, Y. V.</dc:creator>
<dc:creator>Mullen, A. C.</dc:creator>
<dc:date>2023-07-30</dc:date>
<dc:identifier>doi:10.1101/2023.07.29.551032</dc:identifier>
<dc:title><![CDATA[Conserved long noncoding RNA TILAM promotes liver fibrosis through interaction with PML in hepatic stellate cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.31.551045v1?rss=1">
<title>
<![CDATA[
EZH2 inhibition sensitizes IDH1R132H mutant gliomas to histone deacetylase inhibitor. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.31.551045v1?rss=1</link>
<description><![CDATA[
Isocitrate Dehydrogenase-1 (IDH1) is commonly mutated in lower grade diffuse gliomas. The IDH1R132H mutation is an important diagnostic tool for tumor diagnosis and prognosis, however its role in glioma development, and its impact on response to therapy, is not fully understood. We developed a murine model of proneural IDH1R132H mutated glioma that shows elevated production of 2-Hydroxyglutarate (2-HG) and increased tri-methylation of lysine residue K27 on histone H3 (H3K27me3) compared to IDH1 wild-type tumors. We found that using Tazemetostat to inhibit the methyltransferase for H3K27, Enhancer of Zeste 2 (EZH2), reduced H3K27me3 levels and increased acetylation on H3K27. We also found that, although the histone deacetylase inhibitor (HDACi) Panobinostat was less cytotoxic in IDH1R132H mutated cells (either isolated from murine glioma or oligodendrocyte progenitor cells infected in vitro with a retrovirus expressing IDH1R132H) compared to IDH1-wildtype cells, combination treatment with Tazemetostat is synergistic in both mutant and wildtype models. These findings indicate a novel therapeutic strategy for IDH1-mutated gliomas that targets the specific epigenetic alteration in these tumors.

Main PointsMurine gliomas initiated by the IDH1R132H mutation (in the presence of additional genetic alterations, such as p53 loss and PDGF overexpression) recapitulate the metabolic and transcriptional features of the proneural subtype, as they are characterized by increased 2HG levels, and are enriched for OPC lineage-restricted genes compared to IDH-wildtype murine gliomas. In murine IDH1-R132H glioma cells, EZH2 inhibition is not cytotoxic as a monotherapy but reduces levels of H3K27me3 and increases levels of H3K27ac. IDH1R132H cells are relatively resistant to Panobinostat cytotoxicity compared to IDH-wildtype cells, but combining treatment with EZH2 inhibition synergistically kills glioma cells and increases H3K27ac.
]]></description>
<dc:creator>Sprinzen, L.</dc:creator>
<dc:creator>Garcia, F. G.</dc:creator>
<dc:creator>Mela, A.</dc:creator>
<dc:creator>Lei, L.</dc:creator>
<dc:creator>Upadhyayula, P.</dc:creator>
<dc:creator>Mahajan, A.</dc:creator>
<dc:creator>Humala, N.</dc:creator>
<dc:creator>Manier, L.</dc:creator>
<dc:creator>Caprioli, R. M.</dc:creator>
<dc:creator>Quinones-Hinojosa, A.</dc:creator>
<dc:creator>Casaccia, P.</dc:creator>
<dc:creator>Canoll, P.</dc:creator>
<dc:date>2023-08-02</dc:date>
<dc:identifier>doi:10.1101/2023.07.31.551045</dc:identifier>
<dc:title><![CDATA[EZH2 inhibition sensitizes IDH1R132H mutant gliomas to histone deacetylase inhibitor.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.03.551750v1?rss=1">
<title>
<![CDATA[
Cross-family small GTPase ubiquitination by the intracellular pathogen Legionella pneumophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.03.551750v1?rss=1</link>
<description><![CDATA[
The intracellular bacterial pathogen Legionella pneumophila (L.p.) manipulates eukaryotic host ubiquitination machinery to form its replicative vacuole. While nearly 10% of L.p.s arsenal of [~]330 secreted effector proteins have been biochemically characterized as ubiquitin ligases or deubiquitinases, a comprehensive measure of temporally resolved changes in the endogenous host ubiquitinome during infection has not been undertaken. To elucidate how L.p hijacks ubiquitin signaling within the host cell, we undertook a proteome-wide analysis of changes in protein ubiquitination during infection. We discover that L.p. infection results in increased ubiquitination of host proteins regulating subcellular trafficking and membrane dynamics, most notably 63 of [~]160 mammalian Ras superfamily small GTPases. We determine that these small GTPases predominantly undergo non-degradative monoubiquitination, and link ubiquitination to recruitment to the Legionella-containing vacuole membrane. Finally, we find that the bacterial effectors SidC/SdcA play a central, but likely indirect, role in cross-family small GTPase ubiquitination. This work highlights the extensive reconfiguration of host ubiquitin signaling by bacterial effectors during infection and establishes simultaneous ubiquitination of small GTPases across the Ras superfamily as a novel consequence of L.p. infection. This work positions L.p. as a tool to better understand how small GTPases can be regulated by ubiquitination in uninfected contexts.
]]></description>
<dc:creator>Steinbach, A. M.</dc:creator>
<dc:creator>Bhadkamkar, V. L.</dc:creator>
<dc:creator>Jimenez-Morales, D.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Jang, G. M.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:date>2023-08-03</dc:date>
<dc:identifier>doi:10.1101/2023.08.03.551750</dc:identifier>
<dc:title><![CDATA[Cross-family small GTPase ubiquitination by the intracellular pathogen Legionella pneumophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.02.551695v1?rss=1">
<title>
<![CDATA[
Ciliary ARL13B prevents obesity in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.02.551695v1?rss=1</link>
<description><![CDATA[
Cilia are near ubiquitous cellular appendages critical for cell-to-cell communication and involved in diverse developmental and homeostatic processes. ARL13B is a regulatory GTPase enriched in cilia. We engineered an Arl13b mouse allele, Arl13bV358A, which retains ARL13B biochemical activities but renders ARL13B undetectable in cilia. Surprisingly, these mice are hyperphagic and become obese and insulin resistant. In addition to its GTPase function, ARL13B acts as a guanine nucleotide exchange factor (GEF) for ARL3. To test whether ARL13Bs GEF activity is required to regulate body weight, we analyzed the body weight of mice expressing an ARL13B variant lacking ARL3 GEF activity (Arl13bR79Q). We found no difference in body weight, indicating ARL13B is unlikely to regulate weight via its ARL3 GEF activity. Ciliary ARL13B could control energy homeostasis through a role in development or in adult mice. We induced wildtype ARL13B expression, which localizes to cilia, in 4-week-old Arl13bV358A/V358A mice and found the obesity phenotype and associated metabolic impairments were rescued, consistent with ARL13B regulating homeostatic signaling within cilia in adult mice. These results show that ciliary ARL13B functions to control body weight. Our ability to genetically control the subcellular localization of ARL13B by removing and introducing it into cilia enables us to define the cilia-specific role of ARL13B and provides key information for understanding how cilia act as a signaling hub critical for energy homeostasis.

Author SummaryPrimary cilia are essential for energy homeostasis, and their disruption leads to syndromic obesity. However, the mechanisms by which ciliary components regulate energy balance remain unclear. Here, we identify a key role for the ciliary GTPase ARL13B in energy homeostasis. Using a mouse model expressing the ARL13BV358A variant, which is excluded from cilia but retains biochemical activity, we show that ciliary ARL13B regulates body weight, as Arl13bV358A/V358A mice become obese and hyperphagic. Remarkably, restoring ciliary ARL13B in these mice rescues obesity, demonstrating its crucial role in acute ciliary signaling for energy balance. This study directly links ciliary ARL13B with energy balance in adult animals.
]]></description>
<dc:creator>Terry, T. T.</dc:creator>
<dc:creator>Gigante, E. D.</dc:creator>
<dc:creator>Alexandre, C. M.</dc:creator>
<dc:creator>Brewer, K. M.</dc:creator>
<dc:creator>Engle, S. E.</dc:creator>
<dc:creator>Yue, X.</dc:creator>
<dc:creator>Berbari, N. F.</dc:creator>
<dc:creator>Vaisse, C.</dc:creator>
<dc:creator>Caspary, T.</dc:creator>
<dc:date>2023-08-04</dc:date>
<dc:identifier>doi:10.1101/2023.08.02.551695</dc:identifier>
<dc:title><![CDATA[Ciliary ARL13B prevents obesity in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.04.552008v1?rss=1">
<title>
<![CDATA[
Cell growth and nutrient availability control the mitotic exit signaling network in budding yeast 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.04.552008v1?rss=1</link>
<description><![CDATA[
Cell growth is required for cell cycle progression. The amount of growth required for cell cycle progression is reduced in poor nutrients, which leads to a reduction in cell size. In budding yeast, nutrients influence cell size by modulating the duration and extent of bud growth, which occurs predominantly in mitosis. However, the mechanisms are unknown. Here, we used mass spectrometry to identify proteins that mediate the effects of nutrients on bud growth. This led to the discovery that nutrients regulate numerous components of the Mitotic Exit Network (MEN), which controls exit from mitosis. A key component of the MEN undergoes gradual multi-site phosphorylation during bud growth that is dependent upon growth and correlated with the extent of growth. Furthermore, activation of the MEN is sufficient to over-ride a growth requirement for mitotic exit. The data suggest a model in which the MEN integrates signals regarding cell growth and nutrient availability to ensure that mitotic exit occurs only when sufficient growth has occurred.
]]></description>
<dc:creator>Talavera, R. A.</dc:creator>
<dc:creator>Prichard, B. E.</dc:creator>
<dc:creator>Sommer, R. A.</dc:creator>
<dc:creator>Leitao, R. M.</dc:creator>
<dc:creator>Sarabia, C. J.</dc:creator>
<dc:creator>Hazir, S.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Gygi, S.</dc:creator>
<dc:creator>Kellogg, D.</dc:creator>
<dc:date>2023-08-04</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.552008</dc:identifier>
<dc:title><![CDATA[Cell growth and nutrient availability control the mitotic exit signaling network in budding yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.04.552064v1?rss=1">
<title>
<![CDATA[
Using human genetics to develop strategies to increase erythropoietic output from genome-edited hematopoietic stem and progenitor cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.04.552064v1?rss=1</link>
<description><![CDATA[
Human genetic polymorphisms result in a diversity of phenotypes. Some sequences are pathologic and lead to monogenic diseases, while others may confer beneficial traits. Genome editing is a powerful tool to recreate genotypes found in the population, including the ability to correct pathologic mutations. One of the best characterized naturally occurring mutations causing congenital erythrocytosis arises from a truncation in the erythropoietin receptor (tEPOR) which can result in non-pathogenic hyper-production of red blood cells (RBCs). Using the precision of CRISPR/Cas9 genome editing, we have recreated tEPOR and studied the effect of variations of the genotype on RBC development. We then combined tEPOR with a correction strategy developed for {beta}-thalassemia and demonstrated that coupling the two genome editing events gave RBCs a significant selective advantage. This demonstrates the potential of combining human genetics with the precision of genome editing to enable safer and more effective genome editing therapies for patients with serious genetic diseases.
]]></description>
<dc:creator>Camarena, J.</dc:creator>
<dc:creator>Luna, S. E.</dc:creator>
<dc:creator>Hampton, J. P.</dc:creator>
<dc:creator>Majeti, K. R.</dc:creator>
<dc:creator>Charlesworth, C. T.</dc:creator>
<dc:creator>Soupene, E.</dc:creator>
<dc:creator>Sheehan, V. A.</dc:creator>
<dc:creator>Cromer, M. K.</dc:creator>
<dc:creator>Porteus, M. H.</dc:creator>
<dc:date>2023-08-04</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.552064</dc:identifier>
<dc:title><![CDATA[Using human genetics to develop strategies to increase erythropoietic output from genome-edited hematopoietic stem and progenitor cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.04.552066v1?rss=1">
<title>
<![CDATA[
Sulfolobus acidocaldarius adhesion pili power twitching motility in the absence of a dedicated retraction ATPase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.04.552066v1?rss=1</link>
<description><![CDATA[
Type IV pili are ancient and widespread filamentous organelles found in most bacterial and archaeal phyla where they support a wide range of functions, including substrate adhesion, DNA uptake, self aggregation, and cell motility. In most bacteria, PilT-family ATPases disassemble adhesion pili, causing them to rapidly retract and produce twitching motility, important for surface colonization. As archaea do not possess homologs of PilT, it was thought that archaeal pili cannot retract. Here, we employ live-cell imaging under native conditions (75{degrees}C and pH 2), together with automated single-cell tracking, high-temperature fluorescence imaging, and genetic manipulation to demonstrate that S. acidocaldarius exhibits bona fide twitching motility, and that this behavior depends specifically on retractable adhesion pili. Our results demonstrate that archaeal adhesion pili are capable of retraction in the absence of a PilT retraction ATPase and suggests that the ancestral type IV pilus machinery in the last universal common ancestor (LUCA) relied on such a bifunctional ATPase for both extension and retraction.
]]></description>
<dc:creator>Charles-Orszag, A.</dc:creator>
<dc:creator>van Wolferen, M.</dc:creator>
<dc:creator>Lord, S. J.</dc:creator>
<dc:creator>Albers, S.-V.</dc:creator>
<dc:creator>Mullins, D.</dc:creator>
<dc:date>2023-08-04</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.552066</dc:identifier>
<dc:title><![CDATA[Sulfolobus acidocaldarius adhesion pili power twitching motility in the absence of a dedicated retraction ATPase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.02.551383v1?rss=1">
<title>
<![CDATA[
A fibroblast-derived TGFβ/sFRP2 noncanonical Wnt signaling axis underlies epithelial metaplasia in idiopathic pulmonary fibrosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.02.551383v1?rss=1</link>
<description><![CDATA[
Reciprocal interactions between alveolar fibroblasts and epithelial cells are crucial for lung homeostasis, injury repair, and fibrogenesis, but underlying mechanisms remain unclear. To investigate this, we administered the fibroblast-selective TGF{beta}1 signaling inhibitor, epigallocatechin gallate (EGCG), to Interstitial Lung Disease (ILD) patients undergoing diagnostic lung biopsy and conducted single-cell RNA sequencing on spare tissue. Unexposed biopsy samples showed higher fibroblast TGF{beta}1 signaling compared to non-disease donor or end-stage ILD tissues. In vivo, EGCG significantly downregulated TGF{beta}1 signaling and several pro-inflammatory and stress pathways in biopsy samples. Notably, EGCG reduced fibroblast secreted Frizzle-like Receptor Protein 2 (sFRP2), an unrecognized TGF{beta}1 fibroblast target gene induced near type II alveolar epithelial cells (AEC2s). In human AEC2-fibroblast coculture organoids, sFRP2 was essential for AEC2 trans-differentiation to basal cells. Precision cut lung slices (PCLS) from normal donors demonstrated that TGF{beta}1 promoted KRT17 expression and AEC2 morphological change, while sFRP2 was necessary for KRT5 expression in AEC2-derived basaloid cells. Wnt-receptor Frizzled 5 (Fzd5) expression and downstream calcineurin-related signaling in AEC2s were required for sFRP2-induced KRT5 expression. These findings highlight stage-specific TGF{beta}1 signaling in ILD, the therapeutic potential of EGCG in reducing IPF-related transcriptional changes, and identify the TGF{beta}1-non-canonical Wnt pathway crosstalk via sFRP2 as a novel mechanism for dysfunctional epithelial signaling in Idiopathic Pulmonary Fibrosis/ILD.
]]></description>
<dc:creator>Cohen, M. L.</dc:creator>
<dc:creator>Brumwell, A. N.</dc:creator>
<dc:creator>Ho, T. C.</dc:creator>
<dc:creator>Montas, G.</dc:creator>
<dc:creator>Golden, J. A.</dc:creator>
<dc:creator>Jones, K. D.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Chapman, H. A.</dc:creator>
<dc:creator>Le Saux, C. J.</dc:creator>
<dc:date>2023-08-05</dc:date>
<dc:identifier>doi:10.1101/2023.08.02.551383</dc:identifier>
<dc:title><![CDATA[A fibroblast-derived TGFβ/sFRP2 noncanonical Wnt signaling axis underlies epithelial metaplasia in idiopathic pulmonary fibrosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.02.551674v1?rss=1">
<title>
<![CDATA[
Improving rigor and reproducibility in western blot experiments with the blotRig analysis software 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.02.551674v1?rss=1</link>
<description><![CDATA[
Western blot is a popular biomolecular analysis method for measuring the relative quantities of independent proteins in complex biological samples. However, variability in quantitative western blot data analysis poses a challenge in designing reproducible experiments. The lack of rigorous quantitative approaches in current western blot statistical methodology may result in irreproducible inferences. Here we describe best practices for the design and analysis of western blot experiments, with examples and demonstrations of how different analytical approaches can lead to widely varying outcomes. To facilitate best practices, we have developed the blotRig tool for designing and analyzing western blot experiments to improve their rigor and reproducibility. The blotRig application includes functions for counterbalancing experimental design by lane position, batch management across gels, and analytics with covariates and random effects.
]]></description>
<dc:creator>Omondi, C.</dc:creator>
<dc:creator>Chou, A.</dc:creator>
<dc:creator>Fond, K. A.</dc:creator>
<dc:creator>Morioka, K.</dc:creator>
<dc:creator>Joseph, N. R.</dc:creator>
<dc:creator>Sacramento, J. A.</dc:creator>
<dc:creator>Lorio, E.</dc:creator>
<dc:creator>Torres-Espin, A.</dc:creator>
<dc:creator>Radabaugh, H. L.</dc:creator>
<dc:creator>Davis, J. A.</dc:creator>
<dc:creator>Gumbel, J. H.</dc:creator>
<dc:creator>Huie, J. R.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:date>2023-08-05</dc:date>
<dc:identifier>doi:10.1101/2023.08.02.551674</dc:identifier>
<dc:title><![CDATA[Improving rigor and reproducibility in western blot experiments with the blotRig analysis software]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.03.551461v1?rss=1">
<title>
<![CDATA[
A semi-automated method for quantifying optokinetic reflex tracking acuity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.03.551461v1?rss=1</link>
<description><![CDATA[
The study of murine behavioral responses to visual stimuli is a key component of understanding mammalian visual circuitry. One notable response is the optokinetic reflex (OKR), a highly conserved innate behavior necessary for image stabilization on the retina. The OKR provides a robust readout of image tracking ability and has been extensively studied to understand the logic of visual system circuitry and function in mice from different genetic backgrounds. The OKR consists of two phases: a slow tracking phase as the eye follows a stimulus to the edge of the visual plane, and a compensatory fast phase saccade that maintains the image within the visual field. Assessment of the OKR has previously relied on counting individual compensatory eye saccades to estimate tracking speed. To obtain a more direct quantification of tracking ability, we have developed a novel, semi-automated analysis program that allows for rapid and reproducible quantification of unidirectional tracking gains, in addition to being adaptable to any video-oculography equipment. Our analysis program allows for the selection of slow tracking phases, modeling of the vertical and horizontal eye vectors, quantification of eye movement relative to the stimulus, and organization of resultant data into a usable spreadsheet for statistical and graphical comparisons. This quantitative and streamlined analysis pipeline provides a faster and more direct measurement of OKR responses, thereby facilitating further study of visual behavior responses.

SUMMARYWe describe here a semi-automated quantitative analysis method that directly measures eye tracking resulting from murine visual system responses to two-dimensional image motion. A Python-based user interface and analysis algorithm allows for higher throughput and more quantitative measurements of eye tracking parameters than previous methods.
]]></description>
<dc:creator>Kiraly, J. K.</dc:creator>
<dc:creator>Harris, S. C.</dc:creator>
<dc:creator>Al-Khindi, T.</dc:creator>
<dc:creator>Dunn, F.</dc:creator>
<dc:creator>Kolodkin, A. L.</dc:creator>
<dc:date>2023-08-06</dc:date>
<dc:identifier>doi:10.1101/2023.08.03.551461</dc:identifier>
<dc:title><![CDATA[A semi-automated method for quantifying optokinetic reflex tracking acuity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.04.548295v1?rss=1">
<title>
<![CDATA[
Glioma-neuronal circuit remodeling induces regional immunosuppression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.04.548295v1?rss=1</link>
<description><![CDATA[
Neuronal activity-driven mechanisms impact glioblastoma cell proliferation and invasion1-7, and glioblastoma remodels neuronal circuits8,9. Distinct intratumoral regions maintain functional connectivity via a subpopulation of malignant cells that mediate tumor-intrinsic neuronal connectivity and synaptogenesis through their transcriptional programs8. However, the effects of tumor-intrinsic neuronal activity on other cells, such as immune cells, remain unknown. Here we show that regions within glioblastomas with elevated connectivity are characterized by regional immunosuppression. This was accompanied by different cell compositions and inflammatory status of tumor-associated macrophages (TAMs) in the tumor microenvironment. In preclinical intracerebral syngeneic glioblastoma models, CRISPR/Cas9 gene knockout of Thrombospondin-1 (TSP-1/Thbs1), a synaptogenic factor critical for glioma-induced neuronal circuit remodeling, in glioblastoma cells suppressed synaptogenesis and glutamatergic neuronal hyperexcitability, while simultaneously restoring antigen-presentation and pro-inflammatory responses. Moreover, TSP-1 knockout prolonged survival of immunocompetent mice harboring intracerebral syngeneic glioblastoma, but not of immunocompromised mice, and promoted infiltrations of pro-inflammatory TAMs and CD8+ T-cells in the tumor microenvironment. Notably, pharmacological inhibition of glutamatergic excitatory signals redirected tumor-associated macrophages toward a less immunosuppressive phenotype, resulting in prolonged survival. Altogether, our results demonstrate previously unrecognized immunosuppression mechanisms resulting from glioma-neuronal circuit remodeling and suggest future strategies targeting glioma-neuron-immune crosstalk may open up new avenues for immunotherapy.
]]></description>
<dc:creator>Nejo, T.</dc:creator>
<dc:creator>Krishna, S.</dc:creator>
<dc:creator>Jimenez, C.</dc:creator>
<dc:creator>Yamamichi, A.</dc:creator>
<dc:creator>Young, J. S.</dc:creator>
<dc:creator>Lakshmanachetty, S.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Phyu, S. S. S.</dc:creator>
<dc:creator>Ogino, H.</dc:creator>
<dc:creator>Watchmaker, P.</dc:creator>
<dc:creator>Diebold, D.</dc:creator>
<dc:creator>Choudhury, A.</dc:creator>
<dc:creator>Daniel, A. G. S.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:creator>Hervey-Jumper, S. L.</dc:creator>
<dc:creator>Okada, H.</dc:creator>
<dc:date>2023-08-06</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.548295</dc:identifier>
<dc:title><![CDATA[Glioma-neuronal circuit remodeling induces regional immunosuppression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.03.551881v1?rss=1">
<title>
<![CDATA[
BAC Transgenic Expression of Human TREM2-R47H Remodels Amyloid Plaques but Unable to Reprogram Plaque-associated Microglial Reactivity in 5xFAD Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.03.551881v1?rss=1</link>
<description><![CDATA[
BackgroundGenetic study of late-onset Alzheimers disease (AD) reveals that a rare Arginine-to-Histamine mutation at amino acid residue 47 (R47H) in Triggering Receptor Expressed on Myeloid Cells 2 (TREM2) results in increased disease risk. TREM2 plays critical roles in regulating microglial response to amyloid plaques in AD, leading to their clustering and activation surrounding the plaques. We previously showed that increasing human TREM2 gene dosage exerts neuroprotective effects against AD-related deficits in amyloid depositing mouse models of AD. However, the in vivo effects of the R47H mutation on human TREM2-mediated microglial reprogramming and neuroprotection remains poorly understood.

MethodHere we created a BAC transgenic mouse model expressing human TREM2 with the R47H mutation in its cognate genomic context (BAC-TREM2-R47H). Importantly, the BAC used in this study was engineered to delete critical exons of other TREM-like genes on the BAC to prevent confounding effects of overexpressing multiple TREM-like genes. We crossed BAC-TREM2- R47H mice with 5xFAD [1], an amyloid depositing mouse model of AD, to evaluate amyloid pathologies and microglial phenotypes, transcriptomics and in situ expression of key TREM2-dosage dependent genes. We also compared the key findings in 5xFAD/BAC-TREM2-R47H to those observed in 5xFAD/BAC-TREM2 mice.

ResultBoth BAC-TREM2 and BAC-TREM2-R47H showed proper expression of three splicing isoforms of TREM2 that are normally found in human. In 5xFAD background, elevated TREM2-R47H gene dosages significantly reduced the plaque burden, especially the filamentous type. The results were consistent with enhanced phagocytosis and altered NLRP3 inflammasome activation in BAC- TREM2-R47H microglia in vitro. However, unlike TREM2 overexpression, elevated TREM2- R47H in 5xFAD failed to ameliorate cognitive and transcriptomic deficits. In situ analysis of key TREM2-dosage dependent genes and microglial morphology uncovered that TREM2-R47H showed a loss-of-function phenotype in reprogramming of plaque-associated microglial reactivity and gene expression in 5xFAD.

ConclusionOur study demonstrated that the AD-risk variant has a previously unknown, mixture of partial and full loss of TREM2 functions in modulating microglial response in AD mouse brains. Together, our new BAC-TREM2-R47H model and prior BAC-TREM2 mice are invaluable resource to facilitate the therapeutic discovery that target human TREM2 and its R47H variant to ameliorate AD and other neurodegenerative disorders.
]]></description>
<dc:creator>Lee, C. Y. D.</dc:creator>
<dc:creator>De La Rocha, A. J.</dc:creator>
<dc:creator>Inouye, K.</dc:creator>
<dc:creator>Langfelder, P.</dc:creator>
<dc:creator>Daggett, A.</dc:creator>
<dc:creator>Gu, X.</dc:creator>
<dc:creator>Jiang, L.-L.</dc:creator>
<dc:creator>Pamonag, Z.</dc:creator>
<dc:creator>Vaca, R. G.</dc:creator>
<dc:creator>Richman, J.</dc:creator>
<dc:creator>Kawaguchi, R.</dc:creator>
<dc:creator>Gao, F.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Yang, X. W.</dc:creator>
<dc:date>2023-08-07</dc:date>
<dc:identifier>doi:10.1101/2023.08.03.551881</dc:identifier>
<dc:title><![CDATA[BAC Transgenic Expression of Human TREM2-R47H Remodels Amyloid Plaques but Unable to Reprogram Plaque-associated Microglial Reactivity in 5xFAD Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.04.552012v1?rss=1">
<title>
<![CDATA[
Limitations of fluorescent timer proteins toward isolation of developmentally and transcriptionally synchronized populations of human pluripotent stem cells and neural progenitors highlight unique expression and fluorescence maturation kinetics by human cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.04.552012v1?rss=1</link>
<description><![CDATA[
Differentiation of human pluripotent stem cells (hPSC) into distinct neuronal populations holds substantial potential for disease modeling in vitro, toward both elucidation of pathobiological mechanisms and screening of potential therapeutic agents. For successful differentiation of hPSCs into subtype-specific neurons using in vitro protocols, detailed understanding of the transcriptional networks and their dynamic programs regulating endogenous cell fate decisions is critical. One major roadblock is the heterochronic nature of neurodevelopment, during which distinct cells and cell types in the brain and during in vitro differentiation mature and acquire their fates in an unsynchronized manner, hindering pooled transcriptional comparisons. One potential approach is to "translate" chronologic time into linear developmental and maturational time. Attempts to partially achieve this using simple binary promotor-driven fluorescent proteins (FPs) to pool similar cells have not been able to achieve this goal, due to asynchrony of promotor onset in individual cells. Toward solving this, we generated and tested a range of knock-in hPSC lines that express five distinct dual FP timer systems or single time-resolved fluorescent timer (FT) molecules, either in 293T cells or in human hPSCs driving expression from the endogenous paired box 6 (PAX6) promoter of cerebral cortex progenitors. While each of these dual FP or FT systems faithfully reported chronologic time when expressed from a strong inducible promoter in 293T cells, none of the tested FP/FT constructs followed the same fluorescence kinetics in developing human neural progenitor cells, and were unsuccessful in identification and isolation of distinct, developmentally synchronized cortical progenitor populations based on ratiometric fluorescence. This work highlights unique and often surprising expression kinetics and regulation in specific cell types differentiating from hPSCs.
]]></description>
<dc:creator>Peter, M.</dc:creator>
<dc:creator>Shipman, S.</dc:creator>
<dc:creator>Macklis, J. D.</dc:creator>
<dc:date>2023-08-07</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.552012</dc:identifier>
<dc:title><![CDATA[Limitations of fluorescent timer proteins toward isolation of developmentally and transcriptionally synchronized populations of human pluripotent stem cells and neural progenitors highlight unique expression and fluorescence maturation kinetics by human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.04.552017v1?rss=1">
<title>
<![CDATA[
Epigenetic neural glioblastoma enhances synaptic integration and predicts therapeutic vulnerability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.04.552017v1?rss=1</link>
<description><![CDATA[
Neural-tumor interactions drive glioma growth as evidenced in preclinical models, but clinical validation is nascent. We present an epigenetically defined neural signature of glioblastoma that independently affects patients survival. We use reference signatures of neural cells to deconvolve tumor DNA and classify samples into low- or high-neural tumors. High-neural glioblastomas exhibit hypomethylated CpG sites and upregulation of genes associated with synaptic integration. Single-cell transcriptomic analysis reveals high abundance of stem cell-like malignant cells classified as oligodendrocyte precursor and neural precursor cell-like in high-neural glioblastoma. High-neural glioblastoma cells engender neuron-to-glioma synapse formation in vitro and in vivo and show an unfavorable survival after xenografting. In patients, a high-neural signature associates with decreased survival as well as increased functional connectivity and can be detected via DNA analytes and brain-derived neurotrophic factor in plasma. Our study presents an epigenetically defined malignant neural signature in high-grade gliomas that is prognostically relevant.
]]></description>
<dc:creator>Drexler, R.</dc:creator>
<dc:creator>Khatri, R.</dc:creator>
<dc:creator>Sauvigny, T.</dc:creator>
<dc:creator>Mohme, M.</dc:creator>
<dc:creator>Maire, C.</dc:creator>
<dc:creator>Ryba, A.</dc:creator>
<dc:creator>Zghaibeh, Y.</dc:creator>
<dc:creator>Duhrsen, L.</dc:creator>
<dc:creator>Salviano-Silva, A.</dc:creator>
<dc:creator>Lamszus, K.</dc:creator>
<dc:creator>Westphal, M.</dc:creator>
<dc:creator>Gempt, J.</dc:creator>
<dc:creator>Wefers, A. K.</dc:creator>
<dc:creator>Neumann, J.</dc:creator>
<dc:creator>Bode, H.</dc:creator>
<dc:creator>Hausmann, F.</dc:creator>
<dc:creator>Huber, T. B.</dc:creator>
<dc:creator>Bonn, S.</dc:creator>
<dc:creator>Jutten, K.</dc:creator>
<dc:creator>Delev, D.</dc:creator>
<dc:creator>Weber, K. J.</dc:creator>
<dc:creator>Harter, P. N.</dc:creator>
<dc:creator>Onken, J.</dc:creator>
<dc:creator>Vajkoczy, P.</dc:creator>
<dc:creator>Capper, D.</dc:creator>
<dc:creator>Wiestler, B.</dc:creator>
<dc:creator>Weller, M.</dc:creator>
<dc:creator>Snijder, B.</dc:creator>
<dc:creator>Buck, A.</dc:creator>
<dc:creator>Weiss, T.</dc:creator>
<dc:creator>Keough, M. B.</dc:creator>
<dc:creator>Ni, L.</dc:creator>
<dc:creator>Monje, M.</dc:creator>
<dc:creator>Silverbush, D.</dc:creator>
<dc:creator>Hovestadt, V.</dc:creator>
<dc:creator>Suva, M. L.</dc:creator>
<dc:creator>Krishna, S.</dc:creator>
<dc:creator>Hervey-Jumper, S.</dc:creator>
<dc:creator>Schueller, U.</dc:creator>
<dc:creator>Heiland, H. D.</dc:creator>
<dc:creator>Haenzelmann, S.</dc:creator>
<dc:creator>Ricklefs, F. L.</dc:creator>
<dc:date>2023-08-07</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.552017</dc:identifier>
<dc:title><![CDATA[Epigenetic neural glioblastoma enhances synaptic integration and predicts therapeutic vulnerability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.07.552282v1?rss=1">
<title>
<![CDATA[
Two distinct mechanisms of Plexin A function in Drosophila optic lobe lamination and morphogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.07.552282v1?rss=1</link>
<description><![CDATA[
Visual circuit development is characterized by subdivision of neuropils into layers that house distinct sets of synaptic connections. We find that in the Drosophila medulla, this layered organization depends on the axon guidance regulator Plexin A. In plexin A null mutants, synaptic layers of the medulla neuropil and arborizations of individual neurons are wider and less distinct than in controls. Analysis of Semaphorin function indicates that Semaphorin 1a, provided by cells that include Tm5 neurons, is the primary partner for Plexin A in medulla lamination. Removal of the cytoplasmic domain of endogenous Plexin A does not disrupt the formation of medulla layers; however, both null and cytoplasmic domain deletion mutations of plexin A result in an altered overall shape of the medulla neuropil. These data suggest that Plexin A acts as a receptor to mediate morphogenesis of the medulla neuropil, and as a ligand for Semaphorin 1a to subdivide it into layers. Its two independent functions illustrate how a few guidance molecules can organize complex brain structures by each playing multiple roles.

Summary statementThe axon guidance molecule Plexin A has two functions in Drosophila medulla development; morphogenesis of the neuropil requires its cytoplasmic domain, but establishing synaptic layers through Semaphorin 1a does not.
]]></description>
<dc:creator>Bustillo, M. E.</dc:creator>
<dc:creator>Douthit, J.</dc:creator>
<dc:creator>Astigarraga, S.</dc:creator>
<dc:creator>Treisman, J. E.</dc:creator>
<dc:date>2023-08-07</dc:date>
<dc:identifier>doi:10.1101/2023.08.07.552282</dc:identifier>
<dc:title><![CDATA[Two distinct mechanisms of Plexin A function in Drosophila optic lobe lamination and morphogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.07.552354v1?rss=1">
<title>
<![CDATA[
The actin binding protein profilin 1 is critical for mitochondria function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.07.552354v1?rss=1</link>
<description><![CDATA[
Profilin 1 (PFN1) is an actin binding protein that is vital for the polymerization of monomeric actin into filaments. Here we screened knockout cells for novel functions of PFN1 and discovered that mitophagy, a type of selective autophagy that removes defective or damaged mitochondria from the cell, was significantly upregulated in the absence of PFN1. Despite successful autophagosome formation and fusion with the lysosome, and activation of additional mitochondrial quality control pathways, PFN1 knockout cells still accumulate damaged, dysfunctional mitochondria. Subsequent imaging and functional assays showed that loss of PFN1 significantly affects mitochondria morphology, dynamics, and respiration. Further experiments revealed that PFN1 is located to the mitochondria matrix and is likely regulating mitochondria function from within rather than through polymerizing actin at the mitochondria surface. Finally, PFN1 mutants associated with amyotrophic lateral sclerosis (ALS) fail to rescue PFN1 knockout mitochondrial phenotypes and form aggregates within mitochondria, further perturbing them. Together, these results suggest a novel function for PFN1 in regulating mitochondria and identify a potential pathogenic mechanism of ALS-linked PFN1 variants.
]]></description>
<dc:creator>Read, T.-A.</dc:creator>
<dc:creator>Cisterna, B. A.</dc:creator>
<dc:creator>Skruber, K.</dc:creator>
<dc:creator>Ahmadieh, S.</dc:creator>
<dc:creator>Lindamood, H. L.</dc:creator>
<dc:creator>Vitriol, J.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Lefebvre, A. E. Y. T.</dc:creator>
<dc:creator>Black, J. B.</dc:creator>
<dc:creator>Butler, M. T.</dc:creator>
<dc:creator>Bear, J. E.</dc:creator>
<dc:creator>Cherezova, A.</dc:creator>
<dc:creator>Ilatovskaya, D. V.</dc:creator>
<dc:creator>Weintraub, N. L.</dc:creator>
<dc:creator>Vitriol, E. A.</dc:creator>
<dc:date>2023-08-07</dc:date>
<dc:identifier>doi:10.1101/2023.08.07.552354</dc:identifier>
<dc:title><![CDATA[The actin binding protein profilin 1 is critical for mitochondria function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.07.552338v1?rss=1">
<title>
<![CDATA[
Adaptor Protein-3 Produces Synaptic Vesicles that Release Phasic Dopamine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.07.552338v1?rss=1</link>
<description><![CDATA[
The burst firing of midbrain dopamine neurons releases a phasic dopamine signal that mediates reinforcement learning. At many synapses, however, high firing rates deplete synaptic vesicles (SVs), resulting in synaptic depression that limits release. What accounts for the increased release of dopamine by stimulation at high frequency? We find that adaptor protein-3 (AP-3) and its coat protein VPS41 promote axonal dopamine release by targeting vesicular monoamine transporter VMAT2 to the axon rather than dendrites. AP-3 and VPS41 also produce SVs that respond preferentially to high frequency stimulation, independent of their role in axonal polarity. In addition, conditional inactivation of VPS41 in dopamine neurons impairs reinforcement learning, and this involves a defect in the frequency dependence of release rather than the amount of dopamine released. Thus, AP-3 and VPS41 promote the axonal polarity of dopamine release but enable learning by producing a novel population of SVs tuned specifically to high firing frequency that confers the phasic release of dopamine.

Significance statementReinforcement learning requires the phasic dopamine produced by burst firing but synaptic vesicle depletion limits the ability to convey information at high firing rates. We now find that AP-3 has two independent roles in dopamine release. First, AP-3 confers the axonal polarity of dopamine release by targeting vesicular monoamine transporter 2 (VMAT2) to the axon. Second, AP-3 acting locally at the nerve terminal produces synaptic vesicles that respond specifically to high frequency stimulation. Consistent with this, loss of AP-3 impairs reinforcement learning and this reflects the defect in release at high frequency, not the reduction in axonal dopamine.
]]></description>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Yee, A. G.</dc:creator>
<dc:creator>Maas, J.</dc:creator>
<dc:creator>Gierok, S.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Stansil, J.</dc:creator>
<dc:creator>Eriksen, J.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Silm, K.</dc:creator>
<dc:creator>Ford, C. P.</dc:creator>
<dc:creator>Edwards, R. H.</dc:creator>
<dc:date>2023-08-08</dc:date>
<dc:identifier>doi:10.1101/2023.08.07.552338</dc:identifier>
<dc:title><![CDATA[Adaptor Protein-3 Produces Synaptic Vesicles that Release Phasic Dopamine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.08.552451v1?rss=1">
<title>
<![CDATA[
Common and divergent gene regulatory networks control injury-induced and developmental neurogenesis in zebrafish retina. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.08.552451v1?rss=1</link>
<description><![CDATA[
Following acute retinal damage, zebrafish possess the ability to regenerate all neuronal subtypes. This regeneration requires Muller glia (MG) to reprogram and divide asymmetrically to produce a multipotent Muller glia-derived neuronal progenitor cell (MGPC). This raises three key questions. First, does loss of different retinal cell subtypes induce unique MG regeneration responses? Second, do MG reprogram to a developmental retinal progenitor cell state? And finally, to what extent does regeneration recapitulate retinal development? We examined these questions by performing single-nuclear and single-cell RNA-Seq and ATAC-Seq in both developing and regenerating retinas. While MG reprogram to a state similar to late-stage retinal progenitors in developing retinas, there are transcriptional differences between reprogrammed MG/MGPCs and late progenitors, as well as reprogrammed MG in outer and inner retinal damage models. Validation of candidate genes confirmed that loss of different subtypes induces differences in transcription factor gene expression and regeneration outcomes. This work identifies major differences between gene regulatory networks activated following the selective loss of different subtypes of retina neurons, as well as between retinal regeneration and development.
]]></description>
<dc:creator>Lyu, P.</dc:creator>
<dc:creator>Iribarne, M.</dc:creator>
<dc:creator>Serjanov, D.</dc:creator>
<dc:creator>Zhai, Y.</dc:creator>
<dc:creator>Hoang, T.</dc:creator>
<dc:creator>Campbell, L.</dc:creator>
<dc:creator>Boyd, P.</dc:creator>
<dc:creator>Palazzo, I.</dc:creator>
<dc:creator>Nagashima, M.</dc:creator>
<dc:creator>Silva, N. J.</dc:creator>
<dc:creator>Hitchcock, P.</dc:creator>
<dc:creator>Qian, J.</dc:creator>
<dc:creator>Hyde, D.</dc:creator>
<dc:creator>Blackshaw, S.</dc:creator>
<dc:date>2023-08-08</dc:date>
<dc:identifier>doi:10.1101/2023.08.08.552451</dc:identifier>
<dc:title><![CDATA[Common and divergent gene regulatory networks control injury-induced and developmental neurogenesis in zebrafish retina.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.09.552346v1?rss=1">
<title>
<![CDATA[
Environmental challenge rewires functional connections among human genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.09.552346v1?rss=1</link>
<description><![CDATA[
A fundamental question in biology is how a limited number of genes combinatorially govern cellular responses to environmental changes. While the prevailing hypothesis is that relationships between genes, processes, and ontologies could be plastic to achieve this adaptability, quantitatively comparing human gene functional connections between specific environmental conditions at scale is very challenging. Therefore, it remains unclear whether and how human genetic interaction networks are rewired in response to changing environmental conditions. Here, we developed a framework for mapping context-specific genetic interactions, enabling us to measure the plasticity of human genetic architecture upon environmental challenge for [~]250,000 interactions, using cell cycle interruption, genotoxic perturbation, and nutrient deprivation as archetypes. We discover large-scale rewiring of human gene relationships across conditions, highlighted by dramatic shifts in the functional connections of epigenetic regulators (TIP60), cell cycle regulators (PP2A), and glycolysis metabolism. Our study demonstrates that upon environmental perturbation, intra-complex genetic rewiring is rare while inter-complex rewiring is common, suggesting a modular and flexible evolutionary genetic strategy that allows a limited number of human genes to enable adaptation to a large number of environmental conditions.

One Sentence SummaryFive human genetic interaction maps reveal how the landscape of genes functional relationships is rewired as cells experience environmental stress to DNA integrity, cell cycle regulation, and metabolism.
]]></description>
<dc:creator>Herken, B. W.</dc:creator>
<dc:creator>Wong, G.</dc:creator>
<dc:creator>Norman, T.</dc:creator>
<dc:creator>Gilbert, L.</dc:creator>
<dc:date>2023-08-09</dc:date>
<dc:identifier>doi:10.1101/2023.08.09.552346</dc:identifier>
<dc:title><![CDATA[Environmental challenge rewires functional connections among human genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.08.551713v1?rss=1">
<title>
<![CDATA[
Microbiome single cell atlases generated with a commercial instrument 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.08.551713v1?rss=1</link>
<description><![CDATA[
Single cell sequencing is useful for resolving complex systems into their composite cell types and computationally mining them for unique features that are masked in pooled sequencing. However, while commercial instruments have made single cell analysis widespread for mammalian cells, analogous tools for microbes are limited. Here, we present EASi-seq (Easily Accessible Single microbe sequencing). By adapting the single cell workflow of the commercial Mission Bio Tapestri instrument, this method allows for efficient sequencing of individual microbes genomes. EASi-seq allows thousands of microbes to be sequenced per run and, as we show, can generate detailed atlases of human and environmental microbiomes. The ability to capture large shotgun genome datasets from thousands of single microbes provides new opportunities in discovering and analyzing species subpopulations. To facilitate this, we develop a companion bioinformatic pipeline that clusters genome by sequence similarity, improving whole genome assembly, strain identification, taxonomic classification, and gene annotation. In addition, we demonstrate integration of metagenomic contigs with the EASi-seq datasets to reduce capture bias and increase coverage. Overall, EASi-seq enables high quality single cell genomic data for microbiome samples using an accessible workflow that can be run on a commercially available platform.
]]></description>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Demaree, B.</dc:creator>
<dc:creator>Noecker, C.</dc:creator>
<dc:creator>Bisanz, J. E.</dc:creator>
<dc:creator>Weisgerber, D. W.</dc:creator>
<dc:creator>Modavi, C.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:creator>Abate, A. R.</dc:creator>
<dc:date>2023-08-10</dc:date>
<dc:identifier>doi:10.1101/2023.08.08.551713</dc:identifier>
<dc:title><![CDATA[Microbiome single cell atlases generated with a commercial instrument]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.09.551369v1?rss=1">
<title>
<![CDATA[
Plasma Glycomic Markers of Accelerated Biological Aging During Chronic HIV Infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.09.551369v1?rss=1</link>
<description><![CDATA[
People with HIV (PWH) experience an increased vulnerability to premature aging and inflammation-associated comorbidities, even when HIV replication is suppressed by antiretroviral therapy (ART). However, the factors that contribute to or are associated with this vulnerability remain uncertain. In the general population, alterations in the glycomes of circulating IgGs trigger inflammation and precede the onset of aging-associated diseases. Here, we investigate the IgG glycomes of cross-sectional and longitudinal samples from 1,216 women and men, both living with virally suppressed HIV and those without HIV. Our glycan-based machine learning models indicate that living with chronic HIV significantly accelerates the accumulation of pro-aging-associated glycomic alterations. Consistently, PWH exhibit heightened expression of senescence-associated glycan-degrading enzymes compared to their controls. These glycomic alterations correlate with elevated markers of inflammatory aging and the severity of comorbidities, potentially preceding the development of such comorbidities. Mechanistically, HIV-specific antibodies glycoengineered with these alterations exhibit reduced anti-HIV IgG-mediated innate immune functions. These findings hold significant potential for the development of glycomic-based biomarkers and tools to identify and prevent premature aging and comorbidities in people living with chronic viral infections.
]]></description>
<dc:creator>Giron, L. B.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Adeniji, O. S.</dc:creator>
<dc:creator>Yin, X.</dc:creator>
<dc:creator>Kannan, T.</dc:creator>
<dc:creator>Ding, J.</dc:creator>
<dc:creator>Lu, D. Y.</dc:creator>
<dc:creator>Langan, S.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Azevedo, J. L. L. C.</dc:creator>
<dc:creator>Hanna, D. B.</dc:creator>
<dc:creator>Ofotokun, I.</dc:creator>
<dc:creator>Lazar, J.</dc:creator>
<dc:creator>Fischl, M. A.</dc:creator>
<dc:creator>Haberlen, S.</dc:creator>
<dc:creator>Macatangay, B.</dc:creator>
<dc:creator>Adimora, A. A.</dc:creator>
<dc:creator>Jamieson, B. D.</dc:creator>
<dc:creator>Rinaldo, C.</dc:creator>
<dc:creator>Merenstein, D.</dc:creator>
<dc:creator>Roan, N. R.</dc:creator>
<dc:creator>Kutsch, O.</dc:creator>
<dc:creator>Gange, S.</dc:creator>
<dc:creator>Wolinsky, S.</dc:creator>
<dc:creator>Witt, M.</dc:creator>
<dc:creator>Post, W. S.</dc:creator>
<dc:creator>Kossenkov, A.</dc:creator>
<dc:creator>Landay, A.</dc:creator>
<dc:creator>Frank, I.</dc:creator>
<dc:creator>Tien, P. C.</dc:creator>
<dc:creator>Gross, R.</dc:creator>
<dc:creator>Brown, T. T.</dc:creator>
<dc:creator>Mohsen, M. A.</dc:creator>
<dc:date>2023-08-10</dc:date>
<dc:identifier>doi:10.1101/2023.08.09.551369</dc:identifier>
<dc:title><![CDATA[Plasma Glycomic Markers of Accelerated Biological Aging During Chronic HIV Infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.08.552544v1?rss=1">
<title>
<![CDATA[
Interferon response and epigenetic modulation by SMARCA4 mutations drive ovarian tumor immunogenicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.08.552544v1?rss=1</link>
<description><![CDATA[
Cell-intrinsic mechanisms of immunogenicity in ovarian cancer (OC) are not well understood. The presence of damaging mutations in the SWI/SNF chromatin remodeling complex, such as the SMARCA4 (BRG1) catalytic subunit, has been associated with improved response to ICB, however the mechanism by which this occurs is unclear. The aim of this current study was to examine the alterations in tumor cell-intrinsic and extrinsic immune signaling caused by SMARCA4 loss. Using OC models with loss-of-function mutations in SMARCA4, we found that SMARCA4 loss resulted in increased cancer cell-intrinsic immunogenicity, characterized by upregulation of long-terminal RNA repeats such as endogenous retroviruses, increased expression of interferon-stimulated genes, and upregulation of antigen presentation machinery. Notably, this response was dependent on IRF3 signaling, but was independent of the type I interferon receptor. Mice inoculated with cancer cells bearing SMARCA4 loss demonstrated increased activation of cytotoxic T cells and NK cells in the tumor microenvironment as well as increased infiltration with activated dendritic cells. These results were recapitulated when animals bearing SMARCA4-proficient tumors were treated with a BRG1 inhibitor, suggesting that modulation of chromatin remodeling through targeting SMARCA4 may serve as a strategy to reverse immune evasion in OC.
]]></description>
<dc:creator>Brodeur, M. N.</dc:creator>
<dc:creator>Dopeso, H.</dc:creator>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Longhini, A. L. F.</dc:creator>
<dc:creator>Gazzo, A.</dc:creator>
<dc:creator>Sun, S.</dc:creator>
<dc:creator>Koche, R.</dc:creator>
<dc:creator>Qu, R.</dc:creator>
<dc:creator>Hamard, P.-J.</dc:creator>
<dc:creator>Bykov, Y.</dc:creator>
<dc:creator>Green, H.</dc:creator>
<dc:creator>Chiappinelli, K. B.</dc:creator>
<dc:creator>Ozsoy, M. A.</dc:creator>
<dc:creator>Basili, T.</dc:creator>
<dc:creator>Gardner, R.</dc:creator>
<dc:creator>Walderich, S.</dc:creator>
<dc:creator>De Stanchina, E.</dc:creator>
<dc:creator>Greenbaum, B. D.</dc:creator>
<dc:creator>Gonen, M.</dc:creator>
<dc:creator>Weigelt, B.</dc:creator>
<dc:creator>Zamarin, D.</dc:creator>
<dc:date>2023-08-12</dc:date>
<dc:identifier>doi:10.1101/2023.08.08.552544</dc:identifier>
<dc:title><![CDATA[Interferon response and epigenetic modulation by SMARCA4 mutations drive ovarian tumor immunogenicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.08.552553v1?rss=1">
<title>
<![CDATA[
Illuminating the Function of the Orphan Transporter, SLC22A10 in Humans and Other Primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.08.552553v1?rss=1</link>
<description><![CDATA[
SLC22A10 is classified as an orphan transporter with unknown substrates and function. Here we describe the discovery of the substrate specificity and functional characteristics of SLC22A10. The human SLC22A10 tagged with green fluorescent protein was found to be absent from the plasma membrane, in contrast to the SLC22A10 orthologs found in great apes. Estradiol-17{beta}-glucuronide accumulated in cells expressing great ape SLC22A10 orthologs (over 4-fold, p<0.001). In contrast, human SLC22A10 displayed no uptake function. Sequence alignments revealed two amino acid differences including a proline at position 220 of the human SLC22A10 and a leucine at the same position of great ape orthologs. Site-directed mutagenesis yielding the human SLC22A10-P220L produced a protein with excellent plasma membrane localization and associated uptake function. Neanderthal and Denisovan genomes show human-like sequences at proline 220 position, corroborating that SLC22A10 were rendered nonfunctional during hominin evolution after the divergence from the pan lineage (chimpanzees and bonobos). These findings demonstrate that human SLC22A10 is a unitary pseudogene and was inactivated by a missense mutation that is fixed in humans, whereas orthologs in great apes transport sex steroid conjugates.
]]></description>
<dc:creator>Yee, S. W.</dc:creator>
<dc:creator>Ferrandez-Peral, L.</dc:creator>
<dc:creator>Alentorn, P.</dc:creator>
<dc:creator>Fontsere, C.</dc:creator>
<dc:creator>Ceylan, M.</dc:creator>
<dc:creator>Koleske, M. L.</dc:creator>
<dc:creator>Handin, N.</dc:creator>
<dc:creator>Artegoitia, V. M.</dc:creator>
<dc:creator>Lara, G.</dc:creator>
<dc:creator>Chien, H.-C.</dc:creator>
<dc:creator>Zhou, X.</dc:creator>
<dc:creator>Dainat, J.</dc:creator>
<dc:creator>Zalevsky, A.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Brand, C. M.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:creator>Artursson, P.</dc:creator>
<dc:creator>Newman, J. W.</dc:creator>
<dc:creator>Marques-Bonet, T.</dc:creator>
<dc:creator>Giacomini, K. M.</dc:creator>
<dc:date>2023-08-12</dc:date>
<dc:identifier>doi:10.1101/2023.08.08.552553</dc:identifier>
<dc:title><![CDATA[Illuminating the Function of the Orphan Transporter, SLC22A10 in Humans and Other Primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.12.553065v1?rss=1">
<title>
<![CDATA[
PRDM16 co-operates with LHX2 to shape the human brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.12.553065v1?rss=1</link>
<description><![CDATA[
PRDM16 is a dynamic transcriptional regulator of various stem cell niches, including adipocytic, hematopoietic, cardiac progenitors, and neural stem cells. PRDM16 has been suggested to contribute to 1p36 deletion syndrome, one of the most prevalent subtelomeric microdeletion syndromes. We report a patient with a de novo nonsense mutation in the PRDM16 coding sequence, accompanied by lissencephaly and microcephaly features. Human stem cells were genetically modified to mimic this mutation, generating cortical organoids that exhibited altered cell cycle dynamics. RNA sequencing of cortical organoids at day 32 unveiled changes in cell adhesion and WNT-signaling pathways. ChIP-seq of PRDM16 identified binding sites in postmortem human fetal cortex, indicating the conservation of PRDM16 binding to developmental genes in mice and humans, potentially at enhancer sites. A shared motif between PRDM16 and LHX2 was identified and further examined through comparison with LHX2 ChIP-seq data from mice. These results suggested a collaborative partnership between PRDM16 and LHX2 in regulating a common set of genes and pathways in cortical radial glia cells, possibly via their synergistic involvement in cortical development.
]]></description>
<dc:creator>Suresh, V.</dc:creator>
<dc:creator>Bhattacharya, B.</dc:creator>
<dc:creator>Tshuva, R. Y.</dc:creator>
<dc:creator>Danan Gotthold, M.</dc:creator>
<dc:creator>Olender, T.</dc:creator>
<dc:creator>Bose, M.</dc:creator>
<dc:creator>Ben Zeev, B.</dc:creator>
<dc:creator>Smith, R. S.</dc:creator>
<dc:creator>Pradhan, S. J.</dc:creator>
<dc:creator>Tole, S.</dc:creator>
<dc:creator>Galande, S.</dc:creator>
<dc:creator>Harwell, C.</dc:creator>
<dc:creator>Baizabal, M.</dc:creator>
<dc:creator>Reiner, O.</dc:creator>
<dc:date>2023-08-12</dc:date>
<dc:identifier>doi:10.1101/2023.08.12.553065</dc:identifier>
<dc:title><![CDATA[PRDM16 co-operates with LHX2 to shape the human brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.11.551770v1?rss=1">
<title>
<![CDATA[
Flexible regulation of representations on a drifting manifold enables long-term stable complex neuroprosthetic control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.11.551770v1?rss=1</link>
<description><![CDATA[
The nervous system needs to balance the stability of neural representations with plasticity. It is unclear what is the representational stability of simple actions, particularly those that are well-rehearsed in humans, and how it changes in new contexts. Using an electrocorticography brain-computer interface (BCI), we found that the mesoscale manifold and relative representational distances for a repertoire of simple imagined movements were remarkably stable. Interestingly, however, the manifolds absolute location demonstrated day-to-day drift. Strikingly, representational statistics, especially variance, could be flexibly regulated to increase discernability during BCI control without somatotopic changes. Discernability strengthened with practice and was specific to the BCI, demonstrating remarkable contextual specificity. Accounting for drift, and leveraging the flexibility of representations, allowed neuroprosthetic control of a robotic arm and hand for over 7 months without recalibration. Our study offers insight into how electrocorticography can both track representational statistics across long periods and allow long-term complex neuroprosthetic control.
]]></description>
<dc:creator>NATRAJ, N.</dc:creator>
<dc:creator>Seko, S.</dc:creator>
<dc:creator>Abiri, R.</dc:creator>
<dc:creator>Yan, H.</dc:creator>
<dc:creator>Graham, Y.</dc:creator>
<dc:creator>Tu-Chan, A.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Ganguly, K.</dc:creator>
<dc:date>2023-08-14</dc:date>
<dc:identifier>doi:10.1101/2023.08.11.551770</dc:identifier>
<dc:title><![CDATA[Flexible regulation of representations on a drifting manifold enables long-term stable complex neuroprosthetic control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.14.553170v1?rss=1">
<title>
<![CDATA[
Characterization of enhancer activity in early human neurodevelopment using Massively parallel reporter assay (MPRA) and forebrain organoids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.14.553170v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWRegulation of gene expression through enhancers is one of the major processes shaping the structure and function of the human brain during development. High-throughput assays have predicted thousands of enhancers involved in neurodevelopment, and confirming their activity through orthogonal functional assays is crucial. Here, we utilized Massively Parallel Reporter Assays (MPRAs) in stem cells and forebrain organoids to evaluate the activity of [~]7,000 gene-linked enhancers previously identified in human fetal tissues and brain organoids. We used a Gaussian mixture model to evaluate the contribution of background noise in the measured activity signal to confirm the activity of [~]35% of the tested enhancers, with most showing temporal-specific activity, suggesting their evolving role in neurodevelopment. The temporal specificity was further supported by the correlation of activity with gene expression. Our findings provide a valuable gene regulatory resource to the scientific community.

AO_SCPLOWUTHORC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWSUMMARYC_SCPLOWEnhancers are non-coding elements that play a crucial role in the regulation of gene expression during brain development. Despite the availability of various techniques available to identify enhancers, their functional activity is relatively less understood, leaving a gap in our understanding of how enhancer behavior might regulate complex transitions of neurodevelopment. To address this, we utilized forebrain organoids, a 3D model system which closely mimics the complex cellular environment of the developing human brain, and employed Massively Parallel Reporter Assay (MPRA) to validate enhancer activity at various stages of forebrain differentiation, from induced pluripotent stem cells (iPSCs) to neuronal progenitors and cortical neurons. Our study provides a comprehensive catalog of over 2,300 enhancers, showcasing their temporal activity profiles during early neuronal development and offering valuable insights into their likely biological functions. This research advances our understanding of enhancer dynamics in brain development and offers new avenues for further investigations in this field.
]]></description>
<dc:creator>Capauto, D.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Wu, F.</dc:creator>
<dc:creator>Norton, S.</dc:creator>
<dc:creator>Mariani, J.</dc:creator>
<dc:creator>Inoue, F.</dc:creator>
<dc:creator>Crawford, G. E.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Abyzov, A.</dc:creator>
<dc:creator>Vaccarino, F. M.</dc:creator>
<dc:date>2023-08-14</dc:date>
<dc:identifier>doi:10.1101/2023.08.14.553170</dc:identifier>
<dc:title><![CDATA[Characterization of enhancer activity in early human neurodevelopment using Massively parallel reporter assay (MPRA) and forebrain organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.13.553133v1?rss=1">
<title>
<![CDATA[
Addressing biases in gene-set enrichment analysis: a case study of Alzheimer's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.13.553133v1?rss=1</link>
<description><![CDATA[
Inferring the driving regulatory programs from comparative analysis of gene expression data is a cornerstone of systems biology. Many computational frameworks were developed to address this problem, including our iPAGE (information-theoretic Pathway Analysis of Gene Expression) toolset that uses information theory to detect non-random patterns of expression associated with given pathways or regulons1. Our recent observations, however, indicate that existing approaches are susceptible to the biases and artifacts that are inherent to most real world annotations. To address this, we have extended our information-theoretic framework to account for specific biases in biological networks using the concept of conditional information. This novel implementation, called pyPAGE, provides an unbiased way for the estimation of the activity of transcriptional and post-transcriptional regulons.

To showcase pyPAGE, we performed a comprehensive analysis of regulatory perturbations that underlie the molecular etiology of Alzheimers disease (AD). pyPAGE successfully recapitulated several known AD-associated gene expression programs. We also discovered several additional regulons whose differential activity is significantly associated with AD. We further explored how these regulators relate to pathological processes in AD through cell-type specific analysis of single cell gene expression datasets.
]]></description>
<dc:creator>Bakulin, A.</dc:creator>
<dc:creator>Teyssier, N. B.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Khoroshkin, M.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2023-08-15</dc:date>
<dc:identifier>doi:10.1101/2023.08.13.553133</dc:identifier>
<dc:title><![CDATA[Addressing biases in gene-set enrichment analysis: a case study of Alzheimer's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.16.553575v1?rss=1">
<title>
<![CDATA[
Phosphorylation of a Cleaved Tau Proteoform at a Single Residue Inhibits Binding to the E3 Ubiquitin Ligase, CHIP 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.16.553575v1?rss=1</link>
<description><![CDATA[
Microtubule-associated protein tau (MAPT/tau) accumulates in a family of neurodegenerative diseases, including Alzheimers disease (AD). In disease, tau is aberrantly modified by post-translational modifications (PTMs), including hyper-phosphorylation. However, it is often unclear which of these PTMs contribute to taus accumulation or what mechanisms might be involved. To explore these questions, we focused on a cleaved proteoform of tau (tauC3), which selectively accumulates in AD and was recently shown to be degraded by its direct binding to the E3 ubiquitin ligase, CHIP. Here, we find that phosphorylation of tauC3 at a single residue, pS416, is sufficient to block its interaction with CHIP. A co-crystal structure of CHIP bound to the C-terminus of tauC3 revealed the mechanism of this clash and allowed design of a mutation (CHIPD134A) that partially restores binding and turnover of pS416 tauC3. We find that pS416 is produced by the known AD-associated kinase, MARK2/Par-1b, providing a potential link to disease. In further support of this idea, an antibody against pS416 co-localizes with tauC3 in degenerative neurons within the hippocampus of AD patients. Together, these studies suggest a discrete molecular mechanism for how phosphorylation at a specific site contributes to accumulation of an important tau proteoform.
]]></description>
<dc:creator>Nadel, C. M.</dc:creator>
<dc:creator>Wucherer, K.</dc:creator>
<dc:creator>Oehler, A.</dc:creator>
<dc:creator>Thwin, A. C.</dc:creator>
<dc:creator>Basu, K.</dc:creator>
<dc:creator>Callahan, M. D.</dc:creator>
<dc:creator>Southworth, D. R.</dc:creator>
<dc:creator>Mordes, D. A.</dc:creator>
<dc:creator>Craik, C. S.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:date>2023-08-16</dc:date>
<dc:identifier>doi:10.1101/2023.08.16.553575</dc:identifier>
<dc:title><![CDATA[Phosphorylation of a Cleaved Tau Proteoform at a Single Residue Inhibits Binding to the E3 Ubiquitin Ligase, CHIP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.16.553581v1?rss=1">
<title>
<![CDATA[
Assembly of SARS-CoV-2 ribonucleosomes by truncated N* variant of the nucleocapsid protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.16.553581v1?rss=1</link>
<description><![CDATA[
The Nucleocapsid (N) protein of SARS-CoV-2 compacts the RNA genome into viral ribonucleoprotein (vRNP) complexes within virions. Assembly of vRNPs is inhibited by phosphorylation of the N protein SR region. Several SARS-CoV-2 variants of concern carry N protein mutations that reduce phosphorylation and enhance the efficiency of viral packaging. Variants of the dominant B.1.1 viral lineage also encode a truncated N protein, termed N* or {Delta}(1-209), that mediates genome packaging despite lacking the N-terminal RNA-binding domain and SR region. Here, we show that {Delta}(1-209) and viral RNA assemble into vRNPs that are remarkably similar in size and shape to those formed with full-length N protein. We show that assembly of {Delta}(1-209) vRNPs requires the leucine-rich helix (LH) of the central disordered region, and that the LH promotes N protein oligomerization. We also find that fusion of a phosphomimetic SR region to {Delta}(1-209) inhibits RNA binding and vRNP assembly. Our results provide new insights into the mechanisms by which RNA binding promotes N protein self-association and vRNP assembly, and how this process is modulated by SR phosphorylation.
]]></description>
<dc:creator>Adly, A.</dc:creator>
<dc:creator>Bi, M.</dc:creator>
<dc:creator>Carlson, C.</dc:creator>
<dc:creator>Syed, A. M.</dc:creator>
<dc:creator>Ciling, A.</dc:creator>
<dc:creator>Doudna, J.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Morgan, D. O.</dc:creator>
<dc:date>2023-08-17</dc:date>
<dc:identifier>doi:10.1101/2023.08.16.553581</dc:identifier>
<dc:title><![CDATA[Assembly of SARS-CoV-2 ribonucleosomes by truncated N* variant of the nucleocapsid protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.16.553547v1?rss=1">
<title>
<![CDATA[
A Hybrid Transistor with Transcriptionally Controlled Computation and Plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.16.553547v1?rss=1</link>
<description><![CDATA[
Organic electrochemical transistors (OECTs) are ideal devices for translating biological signals into electrical readouts and have applications in bioelectronics, biosensing, and neuromorphic computing. Despite their potential, developing programmable and modular methods for living systems to interface with OECTs has proven challenging. Here we describe hybrid OECTs containing the model electroactive bacterium Shewanella oneidensis that enable the transduction of biological computations to electrical responses. Specifically, we fabricated planar p-type OECTs and demonstrated that channel de-doping is driven by extracellular electron transfer (EET) from S. oneidensis. Leveraging this mechanistic understanding and our ability to control EET flux via transcriptional regulation, we used plasmid-based Boolean logic gates to translate biological computation into current changes within the OECT. Finally, we demonstrated EET-driven changes to OECT synaptic plasticity. This work enables fundamental EET studies and OECT- based biosensing and biocomputing systems with genetically controllable and modular design elements.
]]></description>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Ji, X.</dc:creator>
<dc:creator>Graham, A. J.</dc:creator>
<dc:creator>Dundas, C. M.</dc:creator>
<dc:creator>Mahfoud, I. E. M.</dc:creator>
<dc:creator>Tibbett, B. M.</dc:creator>
<dc:creator>Tan, B.</dc:creator>
<dc:creator>Partipilo, G.</dc:creator>
<dc:creator>Dodabalapur, A.</dc:creator>
<dc:creator>Rivnay, J.</dc:creator>
<dc:creator>Keitz, B. K.</dc:creator>
<dc:date>2023-08-18</dc:date>
<dc:identifier>doi:10.1101/2023.08.16.553547</dc:identifier>
<dc:title><![CDATA[A Hybrid Transistor with Transcriptionally Controlled Computation and Plasticity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.17.553732v1?rss=1">
<title>
<![CDATA[
Enlarged Perivascular Spaces are Associated with White Matter Injury, Brain Atrophy, Cognitive Decline and Markers of Inflammation in an Autosomal Dominant Vascular Neurodegenerative Disease (CADASIL) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.17.553732v1?rss=1</link>
<description><![CDATA[
Background and ObjectivesEnlarged perivascular spaces (ePVS) have been previously reported in Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leucoencephalopathy (CADASIL), but their significance and pathophysiology remains unclear. We investigated associations of ePVS with classical imaging measures, cognitive measures and plasma proteins to better understand what ePVS represents in CADASIL and whether radiographic measures of ePVS would be of value in future therapeutic discovery studies for CADASIL.

Methods24 individuals with CADASIL and 24 age and sex matched controls were included. Disease status was determined based on presence of NOTCH3 mutation. Brain imaging measures of white matter hyperintensity (WMH), brain parenchymal fraction (BPF), ePVS volumes, clinical, and cognitive measures, as well as plasma proteomics were used in models. Global ePVS volumes were calculated via a novel, semi-automated pipeline and levels of 7363 proteins were quantified in plasma using the SomaScan assay. The relationship of ePVS with global burden of WMH, brain atrophy, functional status, neurocognitive measures, and plasma proteins were modelled with linear regression models.

ResultsCADASIL and control groups did not exhibit differences in mean ePVS volumes. However, increased ePVS volumes in CADASIL were associated with increased WMH volume ({beta}=0.57, p=0.05), Clinical Dementia Rating (CDR) Sum-of-Boxes score ({beta}=0.49, p=0.04), and decreased brain parenchymal fraction (BPF) ({beta}=-0.03, p=0.10). In interaction term models, the interaction term between CADASIL disease status and ePVS volume was associated with increased WMH volume ({beta}=0.57, p=0.02), Clinical Dementia Rating (CDR) Sum-of-Boxes score ({beta}=0.52, p=0.02), decreased BPF ({beta}=-0.03, p=0.07) and Mini Mental State Examination (MMSE) score ({beta}=-1.49, p=0.03). Proteins positively associated with ePVS volumes were found to be related to leukocyte migration and inflammation, while negatively associated proteins were related to lipid metabolism. Two central hub proteins were identified in protein networks associated with ePVS volumes: CXCL8/IL-8, and CCL2/MCP-1. The levels of CXCL8/IL8 were also associated with increased WMH volume ({beta}=2.44, p < 0.01), and levels of CCL2/MCP-1 were further associated with decreased BPF ({beta}=-0.0007, p < 0.01), MMSE score ({beta}=-0.02, p < 0.01), and increased Trail Making Test B (TRAILB) completion time ({beta}=0.76, p < 0.01). No protein was associated with all 3 studied imaging measures of pathology (BPF,ePVS,WMH).

DiscussionBased on associations uncovered between ePVS volumes and cognitive functions, imaging and plasma proteins, we conclude that ePVS volumes capture pathologies contributing to chronic brain dysfunction and degeneration in CADASIL, with relevance to future clinical trials for novel therapeutic discoveries to prevent decline and injury in individuals carrying NOTCH3 mutations.
]]></description>
<dc:creator>Karvelas, N.</dc:creator>
<dc:creator>Oh, B.</dc:creator>
<dc:creator>Wang, E.</dc:creator>
<dc:creator>Cobigo, Y.</dc:creator>
<dc:creator>Tsuei, T.</dc:creator>
<dc:creator>Fitzsimons, S.</dc:creator>
<dc:creator>Ehrenberg, A.</dc:creator>
<dc:creator>Geschwind, M.</dc:creator>
<dc:creator>Schwartz, D.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Silbert, L.</dc:creator>
<dc:creator>Rosen, H.</dc:creator>
<dc:creator>Elahi, F. M.</dc:creator>
<dc:date>2023-08-19</dc:date>
<dc:identifier>doi:10.1101/2023.08.17.553732</dc:identifier>
<dc:title><![CDATA[Enlarged Perivascular Spaces are Associated with White Matter Injury, Brain Atrophy, Cognitive Decline and Markers of Inflammation in an Autosomal Dominant Vascular Neurodegenerative Disease (CADASIL)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.17.553736v1?rss=1">
<title>
<![CDATA[
Autophagy counters inflammation-driven glycolytic impairment in aging hematopoietic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.17.553736v1?rss=1</link>
<description><![CDATA[
Aging of the hematopoietic system promotes various blood, immune and systemic disorders and is largely driven by hematopoietic stem cell (HSC) dysfunction (1). Autophagy is central for the benefits associated with activation of longevity signaling programs (2), and for HSC function and response to nutrient stress (3,4). With age, a subset of HSCs increases autophagy flux and preserves some regenerative capacity, while the rest fail to engage autophagy and become metabolically overactivated and dysfunctional (4). However, the signals that promote autophagy in old HSCs and the mechanisms responsible for the increased regenerative potential of autophagy-activated old HSCs remain unknown. Here, we demonstrate that autophagy activation is an adaptive survival response to chronic inflammation in the aging bone marrow (BM) niche (5). We find that inflammation impairs glucose metabolism and suppresses glycolysis in aged HSCs through Socs3-mediated impairment of AKT/FoxO-dependent signaling. In this context, we show that inflammation-mediated autophagy engagement preserves functional quiescence by enabling metabolic adaptation to glycolytic impairment. Moreover, we demonstrate that transient autophagy induction via a short-term fasting/refeeding paradigm normalizes glucose uptake and glycolytic flux and significantly improves old HSC regenerative potential. Our results identify inflammation-driven glucose hypometabolism as a key driver of HSC dysfunction with age and establish autophagy as a targetable node to reset old HSC glycolytic and regenerative capacity.

One-Sentence SummaryAutophagy compensates for chronic inflammation-induced metabolic deregulation in old HSCs, and its transient modulation can reset old HSC glycolytic and regenerative capacity.
]]></description>
<dc:creator>Dellorusso, P. V.</dc:creator>
<dc:creator>Proven, M. A.</dc:creator>
<dc:creator>Calero-Nieto, F. J.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Mitchell, C. A.</dc:creator>
<dc:creator>Hartmann, F.</dc:creator>
<dc:creator>Amouzgar, M.</dc:creator>
<dc:creator>Favaro, P.</dc:creator>
<dc:creator>DeVilbiss, A. W.</dc:creator>
<dc:creator>Swann, J. W.</dc:creator>
<dc:creator>Ho, T. T.</dc:creator>
<dc:creator>Zhao, Z. W.</dc:creator>
<dc:creator>Bendall, S. C.</dc:creator>
<dc:creator>Morrison, S. J.</dc:creator>
<dc:creator>Gottgens, B.</dc:creator>
<dc:creator>Passegue, E.</dc:creator>
<dc:date>2023-08-19</dc:date>
<dc:identifier>doi:10.1101/2023.08.17.553736</dc:identifier>
<dc:title><![CDATA[Autophagy counters inflammation-driven glycolytic impairment in aging hematopoietic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.18.553912v1?rss=1">
<title>
<![CDATA[
Consensus prediction of cell type labels with popV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.18.553912v1?rss=1</link>
<description><![CDATA[
Cell-type classification is a crucial step in single-cell analysis. To facilitate this, several methods have been proposed for the task of transferring a cell-type label from an annotated reference atlas to unannotated query data sets. Existing methods for transferring cell-type labels lack proper uncertainty estimation for the resulting annotations, limiting interpretability and usefulness. To address this, we propose popular Vote (popV, https://github.com/YosefLab/popV), an ensemble of prediction models with an ontology-based voting scheme. PopV achieves accurate cell-type labeling and provides effective uncertainty scores. In multiple case studies, popV confidently annotates the majority of cells while highlighting cell populations that are challenging to annotate. This additional step helps to reduce the load of manual inspection, which is often a necessary component of the annotation process, and enables one to focus on the most problematic parts of the annotation, streamlining the overall annotation process.
]]></description>
<dc:creator>Ergen, C.</dc:creator>
<dc:creator>Xing, G.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Jayasuriya, M.</dc:creator>
<dc:creator>McGeever, E.</dc:creator>
<dc:creator>Pisco, A. O.</dc:creator>
<dc:creator>Streets, A. M.</dc:creator>
<dc:creator>Yosef, N.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.18.553912</dc:identifier>
<dc:title><![CDATA[Consensus prediction of cell type labels with popV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.19.553321v1?rss=1">
<title>
<![CDATA[
Designed Endocytosis-Triggering Proteins mediate Targeted Degradation and amplify signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.19.553321v1?rss=1</link>
<description><![CDATA[
Endocytosis and lysosomal trafficking of cell surface receptors can be triggered by interaction with endogenous ligands. Therapeutic approaches such as LYTAC1,2 and KineTAC3, have taken advantage of this to target specific proteins for degradation by fusing modified native ligands to target binding proteins. While powerful, these approaches can be limited by possible competition with the endogenous ligand(s), the requirement in some cases for chemical modification that limits genetic encodability and can complicate manufacturing, and more generally, there may not be natural ligands which stimulate endocytosis through a given receptor. Here we describe general protein design approaches for designing endocytosis triggering binding proteins (EndoTags) that overcome these challenges. We present EndoTags for the IGF-2R, ASGPR, Sortillin, and Transferrin receptors, and show that fusing these tags to proteins which bind to soluble or transmembrane protein leads to lysosomal trafficking and target degradation; as these receptors have different tissue distributions, the different EndoTags could enable targeting of degradation to different tissues. The modularity and genetic encodability of EndoTags enables AND gate control for higher specificity targeted degradation, and the localized secretion of degraders from engineered cells. The tunability and modularity of our genetically encodable EndoTags should contribute to deciphering the relationship between receptor engagement and cellular trafficking, and they have considerable therapeutic potential as targeted degradation inducers, signaling activators for endocytosis-dependent pathways, and cellular uptake inducers for targeted antibody drug and RNA conjugates.
]]></description>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Abedi, M.</dc:creator>
<dc:creator>Ahn, G.</dc:creator>
<dc:creator>Coventry, B.</dc:creator>
<dc:creator>Sappington, I.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Schlichthaerle, T.</dc:creator>
<dc:creator>Zhang, J. Z.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Goreshnik, I.</dc:creator>
<dc:creator>Chiu, C. W.</dc:creator>
<dc:creator>Chazin-Gray, A.</dc:creator>
<dc:creator>Chan, S.</dc:creator>
<dc:creator>Gerben, S.</dc:creator>
<dc:creator>Murray, A.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>O'Neill, J.</dc:creator>
<dc:creator>Yeh, R.</dc:creator>
<dc:creator>Misquith, A.</dc:creator>
<dc:creator>Wolf, A.</dc:creator>
<dc:creator>Tomasovic, L. M.</dc:creator>
<dc:creator>Piraner, D. I.</dc:creator>
<dc:creator>Gonzalez, M. J. D.</dc:creator>
<dc:creator>Bennett, N. R.</dc:creator>
<dc:creator>Venkatesh, P.</dc:creator>
<dc:creator>Ahlrichs, M.</dc:creator>
<dc:creator>Dobbins, C.</dc:creator>
<dc:creator>Yang, W.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Vafeados, D.</dc:creator>
<dc:creator>Mout, R.</dc:creator>
<dc:creator>Shivaei, S.</dc:creator>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Stewart, L.</dc:creator>
<dc:creator>Spangler, J. B.</dc:creator>
<dc:creator>Bernardes, G. J. L.</dc:creator>
<dc:creator>Roybal, K. T.</dc:creator>
<dc:creator>Greisen, P. J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Bertozzi, C.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.19.553321</dc:identifier>
<dc:title><![CDATA[Designed Endocytosis-Triggering Proteins mediate Targeted Degradation and amplify signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.20.554019v1?rss=1">
<title>
<![CDATA[
Harmonic decomposition of spacetime (HADES) framework characterises the spacetime hierarchy of the DMT brain state 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.20.554019v1?rss=1</link>
<description><![CDATA[
The human brain is a complex system, whose activity exhibits flexible and continuous reorganisation across space and time. The decomposition of whole-brain recordings into harmonic modes has revealed a repertoire of gradient-like activity patterns associated with distinct brain functions. However, the way these activity patterns are expressed over time with their changes in various brain states remains unclear. In this study, we develop the Harmonic Decomposition of Spacetime (HADES) framework that characterises how different harmonic modes defined in space are expressed over time, and, as a proof-of-principle, demonstrate the sensitivity and robustness of this approach to specific changes induced by the serotonergic psychedelic N,N-Dimethyltryptamine (DMT) in healthy participants. HADES demonstrates significant decreases in contributions across most low-frequency harmonic modes in the DMT-induced brain state. When normalizing the contributions by condition (DMT and non-DMT), we detect a decrease specifically in the second functional harmonic, which represents the uni- to transmodal functional hierarchy of the brain, supporting the hypothesis that functional hierarchy is changed in psychedelics. Moreover, HADES dynamic spacetime measures of fractional occupancy, life time and latent space provide a precise description of the significant changes of the spacetime hierarchical organization of brain activity in the psychedelic state.
]]></description>
<dc:creator>Vohryzek, J.</dc:creator>
<dc:creator>Cabral, J.</dc:creator>
<dc:creator>Timmermann, C.</dc:creator>
<dc:creator>Atasoy, S.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Nutt, D.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Deco, G.</dc:creator>
<dc:creator>Kringelbach, M. L.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.20.554019</dc:identifier>
<dc:title><![CDATA[Harmonic decomposition of spacetime (HADES) framework characterises the spacetime hierarchy of the DMT brain state]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.20.554039v1?rss=1">
<title>
<![CDATA[
Acquired stress resilience through bacteria-to-nematode horizontal gene transfer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.20.554039v1?rss=1</link>
<description><![CDATA[
Natural selection drives acquisition of organismal resilience traits to protect against adverse environments. Horizontal gene transfer (HGT) is an important evolutionary mechanism for the acquisition of novel traits, including metazoan acquisition of functions in immunity, metabolism, and reproduction via interdomain HGT (iHGT) from bacteria. We report that the nematode gene rml-3, which was acquired by iHGT from bacteria, enables exoskeleton resilience and protection against environmental toxins in C. elegans. Phylogenetic analysis reveals that diverse nematode RML-3 proteins form a single monophyletic clade most highly similar to bacterial enzymes that biosynthesize L-rhamnose to build cell wall polysaccharides. C. elegans rml-3 is regulated in developing seam cells by heat stress and stress-resistant dauer stage. Importantly, rml-3 deficiency impairs cuticle integrity, barrier functions and organismal stress resilience, phenotypes that are rescued by exogenous L-rhamnose. We propose that iHGT of an ancient bacterial rml-3 homolog enables L-rhamnose biosynthesis in nematodes that facilitates cuticle integrity and organismal resilience in adaptation to environmental stresses during evolution. These findings highlight the remarkable contribution of iHGT on metazoan evolution that is conferred by the domestication of bacterial genes.
]]></description>
<dc:creator>Pandey, T.</dc:creator>
<dc:creator>Kalluraya, C.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Guang, S.</dc:creator>
<dc:creator>Daugherty, M. D.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.20.554039</dc:identifier>
<dc:title><![CDATA[Acquired stress resilience through bacteria-to-nematode horizontal gene transfer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.21.554214v1?rss=1">
<title>
<![CDATA[
Microglia mediate contact-independent neuronal pruning via secreted Neuraminidase-3 associated with extracellular vesicles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.21.554214v1?rss=1</link>
<description><![CDATA[
Neurons communicate with each other through electrochemical transmission at synapses. Microglia, the resident immune cells of the central nervous system, can prune these synapses through a variety of contact-dependent and -independent means. Microglial secretion of active sialidase enzymes upon exposure to inflammatory stimuli is one unexplored mechanism of pruning. Recent work from our lab showed that treatment of neurons with bacterial sialidases disrupts neuronal network connectivity. Here, we find that activated microglia secrete Neuraminidase-3 (Neu3) associated with fusogenic extracellular vesicles. Furthermore, we show Neu3 mediates contact-independent pruning of neurons and subsequent disruption of neuronal networks through neuronal glycocalyx remodeling. We observe that NEU3 is transcriptionally upregulated upon exposure to inflammatory stimuli, and that a genetic knock-out of NEU3 abrogates the sialidase activity of inflammatory microglial secretions. Moreover, we demonstrate that Neu3 is associated with a subpopulation of extracellular vesicles, possibly exosomes, that are secreted by microglia upon inflammatory insult. Finally, we demonstrate that Neu3 is both necessary and sufficient to both desialylate neurons and decrease neuronal network connectivity. These results implicate Neu3 in remodeling of the glycocalyx leading to aberrant network-level activity of neurons, with implications in neuroinflammatory diseases such as Parkinsons disease and Alzheimers disease.

Graphical AbstractNeuroinflammation induces secretion of the sialidase Neu3 via extracellular vesicles from microglia that prune neuronal synapses and disrupt neuronal communication.



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]]></description>
<dc:creator>Delaveris, C. S.</dc:creator>
<dc:creator>Wang, C. L.</dc:creator>
<dc:creator>Riley, N. M.</dc:creator>
<dc:creator>Kulkarni, R. U.</dc:creator>
<dc:creator>Bertozzi, C. R.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.21.554214</dc:identifier>
<dc:title><![CDATA[Microglia mediate contact-independent neuronal pruning via secreted Neuraminidase-3 associated with extracellular vesicles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.21.554173v1?rss=1">
<title>
<![CDATA[
Shh from mossy cells contributes to preventing NSC pool depletion after seizure-induced neurogenesis and in aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.21.554173v1?rss=1</link>
<description><![CDATA[
Epileptic seizures induce aberrant neurogenesis from resident neural stem cells (NSCs) in the dentate gyrus of the adult mouse hippocampus, which has been implicated in depletion of the NSC pool and impairment of hippocampal function. However, the mechanisms regulating neurogenesis after seizures remain unknown. Here we demonstrate that Shh from mossy cells is a major source of Shh signaling activity after seizures, by which mossy cells contribute to seizure-induced neurogenesis and maintenance of the NSC pool. Deletion of Shh from mossy cells attenuates seizure-induced neurogenesis. Moreover, in the absence of Shh from mossy cells, NSCs pool are prematurely depleted after seizure-induced proliferation, and NSCs have impaired self-renewal. Likewise, lack of Shh from mossy cells accelerates age-related decline of the NSC pool with accompanying reduction of self-renewal of NSCs outside the context of pathology such as seizures. Together, our findings indicate that Shh from mossy cells is critical to maintain NSCs and to prevent exhaustion from excessive consumption in aging and after seizures.
]]></description>
<dc:creator>Noguchi, H.</dc:creator>
<dc:creator>Arela, J. C.</dc:creator>
<dc:creator>Ngo, T. T.</dc:creator>
<dc:creator>Cocas, L.</dc:creator>
<dc:creator>Pleasure, S.</dc:creator>
<dc:date>2023-08-22</dc:date>
<dc:identifier>doi:10.1101/2023.08.21.554173</dc:identifier>
<dc:title><![CDATA[Shh from mossy cells contributes to preventing NSC pool depletion after seizure-induced neurogenesis and in aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.21.554174v1?rss=1">
<title>
<![CDATA[
Spatiotemporal molecular dynamics of the developing human thalamus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.21.554174v1?rss=1</link>
<description><![CDATA[
The thalamus plays a central coordinating role in the brain. Thalamic neurons are organized into spatially-distinct nuclei, but the molecular architecture of thalamic development is poorly understood, especially in humans. To begin to delineate the molecular trajectories of cell fate specification and organization in the developing human thalamus, we used single cell and multiplexed spatial transcriptomics. Here we show that molecularly-defined thalamic neurons differentiate in the second trimester of human development, and that these neurons organize into spatially and molecularly distinct nuclei. We identify major subtypes of glutamatergic neuron subtypes that are differentially enriched in anatomically distinct nuclei. In addition, we identify six subtypes of GABAergic neurons that are shared and distinct across thalamic nuclei.

One-Sentence SummarySingle cell and spatial profiling of the developing thalamus in the first and second trimester yields molecular mechanisms of thalamic nuclei development.
]]></description>
<dc:creator>Kim, C.</dc:creator>
<dc:creator>Shin, D.</dc:creator>
<dc:creator>Wang, A.</dc:creator>
<dc:creator>Nowakowski, T.</dc:creator>
<dc:date>2023-08-22</dc:date>
<dc:identifier>doi:10.1101/2023.08.21.554174</dc:identifier>
<dc:title><![CDATA[Spatiotemporal molecular dynamics of the developing human thalamus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.24.552964v1?rss=1">
<title>
<![CDATA[
Regulation of Lung Immune Tone by the Gut-Lung Axis via Dietary Fiber, Gut Microbiota, and Short-Chain Fatty Acids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.24.552964v1?rss=1</link>
<description><![CDATA[
Lung immune tone, i.e. the immune state of the lung, can vary between individuals and over a single individuals lifetime, and its basis and regulation in the context of inflammatory responses to injury is poorly understood. The gut microbiome, through the gut-lung axis, can influence lung injury outcomes but how the diet and microbiota affect lung immune tone is also unclear. We hypothesized that lung immune tone would be influenced by the presence of fiber-fermenting short-chain fatty acid (SCFA)-producing gut bacteria. To test this hypothesis, we conducted a fiber diet intervention study followed by lung injury in mice and profiled gut microbiota using 16S sequencing, metabolomics, and lung immune tone. We also studied germ-free mice to evaluate lung immune tone in the absence of microbiota and performed in vitro mechanistic studies on immune tone and metabolic programming of alveolar macrophages exposed to the SCFA propionate (C3). Mice on high-fiber diet were protected from sterile lung injury compared to mice on a fiber-free diet. This protection strongly correlated with lower lung immune tone, elevated propionate levels and enrichment of specific fecal microbiota taxa; conversely, lower levels of SCFAs and an increase in other fatty acid metabolites and bacterial taxa correlated with increased lung immune tone and increased lung injury in the fiber-free group. In vitro, C3 reduced lung alveolar macrophage immune tone (through suppression of IL-1{beta} and IL-18) and metabolically reprogrammed them (switching from glycolysis to oxidative phosphorylation after LPS challenge). Overall, our findings reveal that the gut-lung axis, through dietary fiber intake and enrichment of SCFA-producing gut bacteria, can regulate innate lung immune tone via IL-1{beta} and IL-18 pathways. These results provide a rationale for the therapeutic development of dietary interventions to preserve or enhance specific aspects of host lung immunity.
]]></description>
<dc:creator>Maruyama, D.</dc:creator>
<dc:creator>Liao, W.-I.</dc:creator>
<dc:creator>Tian, X.</dc:creator>
<dc:creator>Bredon, M.</dc:creator>
<dc:creator>Knapp, J.</dc:creator>
<dc:creator>Doan, T.</dc:creator>
<dc:creator>Tat, C.</dc:creator>
<dc:creator>Chassaing, B.</dc:creator>
<dc:creator>Bhargava, A.</dc:creator>
<dc:creator>Sokol, H.</dc:creator>
<dc:creator>Prakash, A.</dc:creator>
<dc:date>2023-08-25</dc:date>
<dc:identifier>doi:10.1101/2023.08.24.552964</dc:identifier>
<dc:title><![CDATA[Regulation of Lung Immune Tone by the Gut-Lung Axis via Dietary Fiber, Gut Microbiota, and Short-Chain Fatty Acids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.25.554789v1?rss=1">
<title>
<![CDATA[
Rationally seeded computational protein design 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.25.554789v1?rss=1</link>
<description><![CDATA[
Computational protein design is advancing rapidly. Here we describe efficient routes to two families of -helical-barrel proteins with central channels that bind small molecules. The designs are seeded by the sequences and structures of defined de novo oligomeric barrel-forming peptides. Adjacent helices are connected using computational loop building. For targets with antiparallel helices, short loops are sufficient. However, targets with parallel helices require longer connectors; namely, an outer layer of helix-turn-helix-turn-helix motifs that are packed onto the barrels computationally. Throughout these pipelines, residues that define open states of the barrels are maintained. This minimises sequence sampling and accelerates routes to successful designs. For each of 6 targets, just 2 - 6 synthetic genes are made for expression in E. coli. On average, 80% express to give soluble monomeric proteins that are characterized fully, including high-resolution structures for most targets that match the seed structures and design models with high accuracy.
]]></description>
<dc:creator>Albanese, K. I.</dc:creator>
<dc:creator>Petrenas, R.</dc:creator>
<dc:creator>Pirro, F.</dc:creator>
<dc:creator>Naudin, E. A.</dc:creator>
<dc:creator>Borucu, U.</dc:creator>
<dc:creator>Dawson, W. M.</dc:creator>
<dc:creator>Scott, D. A.</dc:creator>
<dc:creator>Leggett, G. J.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:creator>Oliver, T.</dc:creator>
<dc:creator>Woolfson, D.</dc:creator>
<dc:date>2023-08-26</dc:date>
<dc:identifier>doi:10.1101/2023.08.25.554789</dc:identifier>
<dc:title><![CDATA[Rationally seeded computational protein design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.25.554891v1?rss=1">
<title>
<![CDATA[
Control of microtubules in neuronal processes by profilin 1 and actomyosin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.25.554891v1?rss=1</link>
<description><![CDATA[
Microtubules, intermediate filaments, and actin are cytoskeletal polymer networks found within the cell. While each has unique functions, all the cytoskeletal elements must work together for cellular mechanics to be fully operative. This is achieved through crosstalk mechanisms whereby the different networks influence each other through signaling pathways and direct interactions. Because crosstalk can be complex, it is possible for perturbations in one cytoskeletal element to affect the others in ways that are difficult to predict. Here we investigated how long-term changes to the actin cytoskeleton affect microtubules and intermediate filaments. Reducing F-actin or actomyosin contractility increased acetylated microtubules and intermediate filament expression, with the effect being significantly more pronounced in neuronal processes. Changes to microtubules were completely reversible if F-actin and myosin activity is restored. Moreover, the altered microtubules in neuronal processes resulting from F-actin depletion caused significant changes to microtubule-based transport, mimicking phenotypes that are linked to neurodegenerative disease. Thus, defects in actin dynamics cause a compensatory response in other cytoskeleton components which profoundly alters cellular function.
]]></description>
<dc:creator>Cisterna, B. A.</dc:creator>
<dc:creator>Skruber, K.</dc:creator>
<dc:creator>Jane, M. L.</dc:creator>
<dc:creator>Camesi, C. I.</dc:creator>
<dc:creator>Nguyen, I. D.</dc:creator>
<dc:creator>Warp, P. V.</dc:creator>
<dc:creator>Black, J. B.</dc:creator>
<dc:creator>Butler, M. T.</dc:creator>
<dc:creator>Bear, J. E.</dc:creator>
<dc:creator>Read, T.-A.</dc:creator>
<dc:creator>Vitriol, E. A.</dc:creator>
<dc:date>2023-08-26</dc:date>
<dc:identifier>doi:10.1101/2023.08.25.554891</dc:identifier>
<dc:title><![CDATA[Control of microtubules in neuronal processes by profilin 1 and actomyosin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.25.554912v1?rss=1">
<title>
<![CDATA[
CaMKII autophosphorylation but not downstream kinase activity is required for synaptic memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.25.554912v1?rss=1</link>
<description><![CDATA[
CaMKII plays a critical role in long-term potentiation (LTP), a well-established model for learning and memory through the enhancement of synaptic transmission. Biochemical studies indicate that CaMKII catalyzes a phosphotransferase (kinase) reaction of both itself (autophosphorylation) and of multiple downstream target proteins. However, whether either type of phosphorylation plays any role in the synaptic enhancing action of CaMKII remains hotly contested. We have designed a series of experiments to define the minimal requirements for the synaptic enhancement by CaMKII. We find that autophosphorylation of T286 and further binding of CaMKII to the GluN2B subunit are required both for initiating LTP and for its maintenance (synaptic memory). Once bound to the NMDA receptor, the synaptic action of CaMKII occurs in the absence of kinase activity. Thus, autophosphorylation, together with binding to the GluN2B subunit, are the only two requirements for CaMKII in synaptic memory.
]]></description>
<dc:creator>CHEN, X.</dc:creator>
<dc:creator>Cai, Q.</dc:creator>
<dc:creator>zhou, j.</dc:creator>
<dc:creator>Pleasure, S. J.</dc:creator>
<dc:creator>Schulman, H.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Nicoll, R. A.</dc:creator>
<dc:date>2023-08-26</dc:date>
<dc:identifier>doi:10.1101/2023.08.25.554912</dc:identifier>
<dc:title><![CDATA[CaMKII autophosphorylation but not downstream kinase activity is required for synaptic memory]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.25.554311v1?rss=1">
<title>
<![CDATA[
Decreased accuracy of forensic DNA mixture analysis for groups with lower genetic diversity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.25.554311v1?rss=1</link>
<description><![CDATA[
Forensic investigation of DNA samples from multiple contributors has become commonplace. These complex analyses use statistical frameworks accounting for multiple levels of uncertainty in allelic contributions from different individuals, particularly for samples containing few molecules of DNA. These methods have been thoroughly tested along some axes of variation, but less attention has been paid to accuracy across human genetic variation. Here, we quantify the accuracy of DNA mixture analysis over 244 human groups. We find higher false inclusion rates for mixtures with more contributors, and for groups with lower genetic diversity. Even for two-contributor mixtures where one contributor is known and the reference group is correctly specified, false inclusion rates are 1e-5 or higher for 56 out of 244 groups. This means that, depending on multiple testing, some false inclusions may be expected. These false positives could be lessened with more selective and conservative use of DNA mixture analysis.

HIGHLIGHTSO_LIGroups with lower genetic diversity have higher mixture analysis false positive rates.
C_LIO_LIAnalyses with mis-specified references have somewhat higher false positive rates.
C_LIO_LIMixture analysis accuracy decreases with more mixture contributors.
C_LI

GRAPHICAL ABSTRACT

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]]></description>
<dc:creator>Flores, M.</dc:creator>
<dc:creator>Ly, C.</dc:creator>
<dc:creator>Ho, E.</dc:creator>
<dc:creator>Ceberio, N.</dc:creator>
<dc:creator>Felix, K.</dc:creator>
<dc:creator>Thorner, H. M.</dc:creator>
<dc:creator>Guardado, M.</dc:creator>
<dc:creator>Paunovich, M.</dc:creator>
<dc:creator>Godek, C.</dc:creator>
<dc:creator>Kalaydjian, C.</dc:creator>
<dc:creator>Rohlfs, R.</dc:creator>
<dc:date>2023-08-27</dc:date>
<dc:identifier>doi:10.1101/2023.08.25.554311</dc:identifier>
<dc:title><![CDATA[Decreased accuracy of forensic DNA mixture analysis for groups with lower genetic diversity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.25.554320v1?rss=1">
<title>
<![CDATA[
S-Nitrosylation of CRTC1 in Alzheimer's disease impairs CREB-dependent gene expression induced by neuronal activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.25.554320v1?rss=1</link>
<description><![CDATA[
CREB-regulated transcription coactivator 1 (CRTC1) plays an important role in synaptic plasticity, learning and long-term memory formation through regulation of neuronal activity-dependent gene expression, and CRTC1 dysregulation is implicated in Alzheimers disease (AD). Here, we show that increased S-nitrosylation of CRTC1 (forming SNO-CRTC1), as seen in cell-based, animal-based, and human induced pluripotent stem cell (hiPSC)-derived cerebrocortical neuron-based AD models, disrupts its binding with CREB and diminishes the activity-dependent gene expression mediated by the CRTC1/CREB pathway. We identified Cys216 of CRTC1 as the primary target of S-nitrosylation by nitric oxide (NO)-related species. Using CRISPR/Cas9 techniques, we mutated Cys216 to Ala in hiPSC-derived cerebrocortical neurons bearing one allele of the APPSwe mutation (AD-hiPSC neurons). Introduction of this non-nitrosylatable CRTC1 construct rescued defects in AD-hiPSC neurons, including decreased neurite length and increased neuronal cell death. Additionally, expression of non-nitrosylatable CRTC1 in vivo in the hippocampus rescued synaptic plasticity in the form of long-term potentiation (LTP) in 5XFAD mice. Taken together, these results demonstrate that formation of SNO-CRTC1 contributes to the pathogenesis of AD by attenuating the neuronal activity-dependent CREB transcriptional pathway, and suggests a novel therapeutic target for AD.
]]></description>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Vlkolinsky, R.</dc:creator>
<dc:creator>Wu, C.</dc:creator>
<dc:creator>Dolatabadi, N.</dc:creator>
<dc:creator>Scott, H.</dc:creator>
<dc:creator>Prikhodko, O.</dc:creator>
<dc:creator>Blanco, M.</dc:creator>
<dc:creator>Lang, N.</dc:creator>
<dc:creator>Pina-Crespo, J.</dc:creator>
<dc:creator>Nakamura, T.</dc:creator>
<dc:creator>Roberto, M.</dc:creator>
<dc:creator>Lipton, S. A.</dc:creator>
<dc:date>2023-08-27</dc:date>
<dc:identifier>doi:10.1101/2023.08.25.554320</dc:identifier>
<dc:title><![CDATA[S-Nitrosylation of CRTC1 in Alzheimer's disease impairs CREB-dependent gene expression induced by neuronal activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.26.554969v1?rss=1">
<title>
<![CDATA[
Identification of genetic drivers of plasma lipoproteins in the Diversity Outbred mouse population 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.26.554969v1?rss=1</link>
<description><![CDATA[
Despite great progress in understanding lipoprotein physiology, there is still much to be learned about the genetic drivers of lipoprotein abundance, composition, and function. We used ion mobility spectrometry to survey 16 plasma lipoprotein subfractions in 500 Diversity Outbred (DO) mice maintained on a Western-style diet. We identified 21 quantitative trait loci (QTL) affecting lipoprotein abundance. To refine the QTL and link them to disease risk in humans, we asked if the human homologues of genes located at each QTL were associated with lipid traits in human genome-wide association studies (GWAS). Integration of mouse QTL with human GWAS yielded candidate gene drivers for 18 of the 21 QTL. This approach enabled us to nominate the gene encoding the neutral ceramidase, Asah2, as a novel candidate driver at a QTL on chromosome 19 for large HDL particles (HDL-2b). To experimentally validate Asah2, we surveyed lipoproteins in Asah2-/-mice. Compared to wild-type mice, female Asah2-/- mice showed an increase in several lipoproteins, including HDL. Our results provide insights into the genetic regulation of circulating lipoproteins, as well as mechanisms by which lipoprotein subfractions may affect cardiovascular disease risk in humans.
]]></description>
<dc:creator>Price, T. R.</dc:creator>
<dc:creator>Emfinger, C. H.</dc:creator>
<dc:creator>Schueler, K. L.</dc:creator>
<dc:creator>King, S.</dc:creator>
<dc:creator>Nicholson, R.</dc:creator>
<dc:creator>Beck, T.</dc:creator>
<dc:creator>Yandell, B. S.</dc:creator>
<dc:creator>Summers, S.</dc:creator>
<dc:creator>Holland, W. L.</dc:creator>
<dc:creator>Krauss, R. M.</dc:creator>
<dc:creator>Keller, M. P.</dc:creator>
<dc:creator>Attie, A. D.</dc:creator>
<dc:date>2023-08-27</dc:date>
<dc:identifier>doi:10.1101/2023.08.26.554969</dc:identifier>
<dc:title><![CDATA[Identification of genetic drivers of plasma lipoproteins in the Diversity Outbred mouse population]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.26.554851v1?rss=1">
<title>
<![CDATA[
Learning chemical sensitivity reveals mechanisms of cellular response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.26.554851v1?rss=1</link>
<description><![CDATA[
Chemical probes interrogate disease mechanisms at the molecular level by linking genetic changes to observable traits. However, comprehensive chemical screens in diverse biological models are impractical. To address this challenge, we developed ChemProbe, a model that predicts cellular sensitivity to hundreds of molecular probes and drugs by learning to combine transcriptomes and chemical structures. Using ChemProbe, we inferred the chemical sensitivity of cancer cell lines and tumor samples and analyzed how the model makes predictions. We retrospectively evaluated drug response predictions for precision breast cancer treatment and prospectively validated chemical sensitivity predictions in new cellular models, including a genetically modified cell line. Our model interpretation analysis identified transcriptome features reflecting compound targets and protein network modules, identifying genes that drive ferroptosis. ChemProbe is an interpretable in silico screening tool that allows researchers to measure cellular response to diverse compounds, facilitating research into molecular mechanisms of chemical sensitivity.
]]></description>
<dc:creator>Connell, W. J.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Keiser, M. J.</dc:creator>
<dc:date>2023-08-28</dc:date>
<dc:identifier>doi:10.1101/2023.08.26.554851</dc:identifier>
<dc:title><![CDATA[Learning chemical sensitivity reveals mechanisms of cellular response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.28.554707v1?rss=1">
<title>
<![CDATA[
Brain-Derived CCN3 Is An Osteoanabolic Hormone That Sustains Bone in Lactating Females 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.28.554707v1?rss=1</link>
<description><![CDATA[
In lactating mothers, the high calcium (Ca2+) demand for milk production triggers significant bone resorption. While estrogen would normally counteract excessive bone loss and maintain sufficient bone formation during this postpartum period, this sex steroid drops precipitously after giving birth. Here, we report that brain-derived CCN3 (Cellular Communication Network factor 3) secreted from KISS1 neurons of the arcuate nucleus (ARCKISS1) fills this void and functions as a potent osteoanabolic factor to promote bone mass in lactating females. Using parabiosis and bone transplant methods, we first established that a humoral factor accounts for the female-specific, high bone mass previously observed by our group after deleting estrogen receptor alpha (ER) from ARCKISS1 neurons1. This exceptional bone phenotype in mutant females can be traced back to skeletal stem cells (SSCs), as reflected by their increased frequency and osteochondrogenic potential. Based on multiple assays, CCN3 emerged as the most promising secreted pro-osteogenic factor from ARCKISS1 neurons, acting on mouse and human SSCs at low subnanomolar concentrations independent of age or sex. That brain-derived CCN3 promotes bone formation was further confirmed by in vivo gain- and loss-of-function studies. Notably, a transient rise in CCN3 appears in ARCKISS1 neurons in estrogen-depleted lactating females coincident with increased bone remodeling and high calcium demand. Our findings establish CCN3 as a potentially new therapeutic osteoanabolic hormone that defines a novel female-specific brain-bone axis for ensuring mammalian species survival.
]]></description>
<dc:creator>Babey, M. E.</dc:creator>
<dc:creator>Krause, W. C.</dc:creator>
<dc:creator>Herber, C. B.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Nikkannen, J.</dc:creator>
<dc:creator>Rodriquez, R.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Castro-Navarro, F.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Villeda, S.</dc:creator>
<dc:creator>Lance, N. E.</dc:creator>
<dc:creator>Scheller, E. L.</dc:creator>
<dc:creator>Chan, C. K. F.</dc:creator>
<dc:creator>Ambrosi, T. H.</dc:creator>
<dc:creator>Ingraham, H. A.</dc:creator>
<dc:date>2023-08-28</dc:date>
<dc:identifier>doi:10.1101/2023.08.28.554707</dc:identifier>
<dc:title><![CDATA[Brain-Derived CCN3 Is An Osteoanabolic Hormone That Sustains Bone in Lactating Females]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.28.555157v1?rss=1">
<title>
<![CDATA[
Tau fibrils induce nanoscale membrane damage and nucleate cytosolic tau at lysosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.28.555157v1?rss=1</link>
<description><![CDATA[
The prion-like spread of protein aggregates is a leading hypothesis for the propagation of neurofibrillary lesions in the brain, including the spread of tau inclusions associated with Alzheimers disease. The mechanisms of cellular uptake of tau seeds and subsequent nucleated polymerization of cytosolic tau are major questions in the field, and the potential for coupling between the entry and nucleation mechanisms has been little explored. We found that in primary astrocytes, endocytosis of tau seeds leads to their accumulation in lysosomes. This in turn leads to lysosomal swelling, deacidification and recruitment of ESCRT proteins, but not Galectin-3, to the lysosomal membrane. These observations are consistent with nanoscale damage of the lysosomal membrane. Using live cell and STORM, imaging, nucleation of cytosolic tau occurs primarily at the lysosome membrane under these conditions. These data suggest that tau seeds escape from lysosomes via nanoscale damage rather than wholesale rupture, and that nucleation of cytosolic tau commences as soon as tau fibril ends emerge from the lysosomal membrane.
]]></description>
<dc:creator>Rose, K.</dc:creator>
<dc:creator>Jepson, T.</dc:creator>
<dc:creator>Shukla, S.</dc:creator>
<dc:creator>Maya-Romero, A.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Xu, K.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2023-08-28</dc:date>
<dc:identifier>doi:10.1101/2023.08.28.555157</dc:identifier>
<dc:title><![CDATA[Tau fibrils induce nanoscale membrane damage and nucleate cytosolic tau at lysosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.28.555153v1?rss=1">
<title>
<![CDATA[
Neuronal apoE4 induces early hyperexcitability in select populations of hippocampal neurons by altering Nell2 expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.28.555153v1?rss=1</link>
<description><![CDATA[
The full impact of apolipoprotein E4 (APOE4), the strongest genetic risk factor for Alzheimers disease (AD), on neuronal and network function remains unclear. We found hippocampal region-specific network hyperexcitability in young APOE4 knock-in (E4-KI) mice which predicted cognitive deficits at old age. Network hyperexcitability in young E4-KI mice was mediated by hippocampal region-specific subpopulations of smaller and hyperexcitable neurons that were eliminated by selective removal of neuronal APOE4. Aged E4-KI mice exhibited hyperexcitable granule cells, a progressive inhibitory deficit, and E/I imbalance in the dentate gyrus, exacerbating hippocampal hyperexcitability. Single-nucleus RNA-sequencing revealed neuronal cell type-specific and age-dependent transcriptomic changes, including Nell2 overexpression in E4-KI mice. Reducing Nell2 expression in specific neuronal types of E4-KI mice with CRISPRi rescued their abnormal excitability phenotypes, implicating Nell2 overexpression as a cause of APOE4-induced hyperexcitability. These findings highlight the early transcriptomic and electrophysiological alterations underlying APOE4-induced hippocampal network dysfunction and its contribution to AD pathogenesis with aging.
]]></description>
<dc:creator>Jang, S.-S.</dc:creator>
<dc:creator>Tabuena, D. R.</dc:creator>
<dc:creator>Grone, B.</dc:creator>
<dc:creator>Yip, O.</dc:creator>
<dc:creator>Blumenfeld, J.</dc:creator>
<dc:creator>Koutsodendris, N.</dc:creator>
<dc:creator>Ding, L.</dc:creator>
<dc:creator>Xu, Q.</dc:creator>
<dc:creator>Yoon, S. Y.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Zilberter, M.</dc:creator>
<dc:date>2023-08-29</dc:date>
<dc:identifier>doi:10.1101/2023.08.28.555153</dc:identifier>
<dc:title><![CDATA[Neuronal apoE4 induces early hyperexcitability in select populations of hippocampal neurons by altering Nell2 expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.28.555217v1?rss=1">
<title>
<![CDATA[
Sex differences in discrimination behavior and orbitofrontal engagement during context-gated reward prediction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.28.555217v1?rss=1</link>
<description><![CDATA[
Animals, including humans, rely on contextual information to interpret ambiguous stimuli. Impaired context processing is a hallmark of several neuropsychiatric disorders, including schizophrenia, autism spectrum disorders, post-traumatic stress disorder, and addiction. While sex differences in the prevalence and manifestations of these disorders are well established, potential sex differences in context processing remain uncertain. Here we examined sex differences in the contextual control over cue-evoked reward seeking and its neural correlates, in rats. Male and female rats were trained in a bidirectional occasion-setting preparation in which the validity of two auditory reward-predictive cues was informed by the presence, or absence, of a visual contextual feature (LIGHT: X+ / DARK: X- / LIGHT: Y- / DARK: Y+). Females were significantly slower to acquire contextual control over cue-evoked reward seeking. However, once established, the contextual control over behavior was more robust in female rats; it showed less within-session variability (less influence of prior reward) and greater resistance to acute stress. This superior contextual control achieved by females was accompanied by an increased activation of the orbitofrontal cortex compared to males. Critically, these behavioral and neural sex differences were specific to the contextual modulation process and not observed in simple, context-independent, reward prediction tasks. These results indicate a sex-biased trade-off between the speed of acquisition and the robustness of performance in the contextual modulation of cued reward seeking. The different distribution of sexes along the fast learning {leftrightarrow} steady performance continuum might reflect different levels of engagement of the orbitofrontal cortex, and might have implications for our understanding of sex differences in psychiatric disorders.
]]></description>
<dc:creator>Peterson, S.</dc:creator>
<dc:creator>Maheras, A.</dc:creator>
<dc:creator>Wu, B.</dc:creator>
<dc:creator>Chavira, J.</dc:creator>
<dc:creator>Keiflin, R.</dc:creator>
<dc:date>2023-08-29</dc:date>
<dc:identifier>doi:10.1101/2023.08.28.555217</dc:identifier>
<dc:title><![CDATA[Sex differences in discrimination behavior and orbitofrontal engagement during context-gated reward prediction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.30.555359v1?rss=1">
<title>
<![CDATA[
Functional characterization of gene regulatory elements and neuropsychiatric disease-associated risk loci in iPSCs and iPSC-derived neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.30.555359v1?rss=1</link>
<description><![CDATA[
Genome-wide association studies (GWAS) have identified thousands of non-coding variants that contribute to psychiatric disease risks, likely by perturbing cis-regulatory elements (CREs). However, our ability to interpret and explore their mechanisms of action is hampered by a lack of annotation of functional CREs (fCREs) in neural cell types. Here, through genome-scale CRISPR screens of 22,000 candidate CREs (cCREs) in human induced pluripotent stem cells (iPSCs) undergoing differentiation to excitatory neurons, we identify 2,847 and 5,540 fCREs essential for iPSC fitness and neuronal differentiation, respectively. These fCREs display dynamic epigenomic features and exhibit increased numbers and genomic spans of chromatin interactions following terminal neuronal differentiation. Furthermore, fCREs essential for neuronal differentiation show significantly greater enrichment of genetic heritability for neurodevelopmental diseases including schizophrenia (SCZ), attention deficit hyperactivity disorder (ADHD), and autism spectrum disorders (ASD) than cCREs. Using high-throughput prime editing screens we experimentally confirm 45 SCZ risk variants that act by affecting the function of fCREs. The extensive and in-depth functional annotation of cCREs in neuronal types therefore provides a crucial resource for interpreting non-coding risk variants of neuropsychiatric disorders.
]]></description>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Jones, I. R.</dc:creator>
<dc:creator>Chen, P. B.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Zheng, L.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Li, Y. E.</dc:creator>
<dc:creator>Sun, Q.</dc:creator>
<dc:creator>Wen, J.</dc:creator>
<dc:creator>Beaman, C.</dc:creator>
<dc:creator>Cui, X.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Hu, M.</dc:creator>
<dc:creator>Ren, B.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:date>2023-08-30</dc:date>
<dc:identifier>doi:10.1101/2023.08.30.555359</dc:identifier>
<dc:title><![CDATA[Functional characterization of gene regulatory elements and neuropsychiatric disease-associated risk loci in iPSCs and iPSC-derived neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.29.555389v1?rss=1">
<title>
<![CDATA[
PRDM6 promotes medulloblastoma by repressing chromatin accessibility and altering gene expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.29.555389v1?rss=1</link>
<description><![CDATA[
SNCAIP duplication may promote Group 4 medulloblastoma via induction of PRDM6, a poorly characterized member of the PRDF1 and RIZ1 homology domain-containing (PRDM) family of transcription factors. Here, we investigated the function of PRDM6 in human hindbrain neuroepithelial stem cells and tested PRDM6 as a driver of Group 4 medulloblastoma. We report that human PRDM6 localizes predominantly to the nucleus, where it causes widespread repression of chromatin accessibility and complex alterations of gene expression patterns. Genome-wide mapping of PRDM6 binding reveals that PRDM6 binds to chromatin regions marked by histone H3 lysine 27 trimethylation that are located within, or proximal to, genes. Moreover, we show that PRDM6 expression in neuroepithelial stem cells promotes medulloblastoma. Surprisingly, medulloblastomas derived from PRDM6-expressing neuroepithelial stem cells match human Group 3, but not Group 4, medulloblastoma. We conclude that PRDM6 expression has oncogenic potential but is insufficient to drive Group 4 medulloblastoma from neuroepithelial stem cells. We propose that both PRDM6 and additional factors, such as specific cell-of-origin features, are required for Group 4 medulloblastoma. Given the lack of PRDM6 expression in normal tissues and its oncogenic potential shown here, we suggest that PRDM6 inhibition may have therapeutic value in PRDM6-expressing medulloblastomas.
]]></description>
<dc:creator>Schmidt, C.</dc:creator>
<dc:creator>Cohen, S.</dc:creator>
<dc:creator>Gudenas, B. L.</dc:creator>
<dc:creator>Husain, S.</dc:creator>
<dc:creator>Carlson, A.</dc:creator>
<dc:creator>Westelman, S.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Phillips, J. J.</dc:creator>
<dc:creator>Northcott, P. A.</dc:creator>
<dc:creator>Weiss, W. A.</dc:creator>
<dc:creator>Schwer, B.</dc:creator>
<dc:date>2023-08-31</dc:date>
<dc:identifier>doi:10.1101/2023.08.29.555389</dc:identifier>
<dc:title><![CDATA[PRDM6 promotes medulloblastoma by repressing chromatin accessibility and altering gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.30.555562v1?rss=1">
<title>
<![CDATA[
The small G-protein Rac1 in the dorsomedial striatum promotes alcohol-dependent structural plasticity and goal-directed learning in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.30.555562v1?rss=1</link>
<description><![CDATA[
The small G-protein Rac1 promotes the formation of filamentous actin (F-Actin). Actin is a major component of dendritic spines, and we previously found that alcohol alters actin composition and dendritic spine structure in the nucleus accumbens (NAc) and the dorsomedial striatum (DMS). To examine if Rac1 contributes to these alcohol-mediated adaptations, we measured the level of GTP-bound active Rac1 in the striatum of mice following 7 weeks of intermittent access to 20% alcohol. We found that chronic alcohol intake activates Rac1 in the DMS of male mice. In contrast, Rac1 is not activated by alcohol in the NAc and DLS of male mice, or in the DMS of female mice. Similarly, closely related small G-proteins are not activated by alcohol in the DMS, and Rac1 activity is not increased in the DMS by moderate alcohol or natural reward. To determine the consequences of alcohol-dependent Rac1 activation in the DMS of male mice, we inhibited endogenous Rac1 by infecting the DMS of mice with an AAV expressing a dominant negative form of the small G-protein (Rac1-DN). We found that overexpression of AAV-Rac1-DN in the DMS inhibits alcohol-mediated Rac1 signaling and attenuates alcohol-mediated F-actin polymerization, which corresponded with a decrease in dendritic arborization and spine maturation. Finally, we provide evidence to suggest that Rac1 in the DMS plays a role in alcohol-associated goal-directed learning. Together, our data suggest that Rac1 in the DMS plays an important role in alcohol-dependent structural plasticity and aberrant learning.

Significance StatementAddiction, including alcohol use disorder, is characterized by molecular and cellular adaptations that promote maladaptive behaviors. We found that Rac1 was activated by alcohol in the dorsomedial striatum (DMS) of male mice. We show that alcohol-mediated Rac1 signaling is responsible for alterations in actin dynamics and neuronal morphology. We also present data to suggest that Rac1 is important for alcohol-associated learning processes. These results suggest that Rac1 in the DMS is an important contributor to adaptations that promote alcohol use disorder.
]]></description>
<dc:creator>Hoisington, Z. W.</dc:creator>
<dc:creator>Salvi, A.</dc:creator>
<dc:creator>Laguesse, S.</dc:creator>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>Shukla, C.</dc:creator>
<dc:creator>Phamluong, K.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2023-09-01</dc:date>
<dc:identifier>doi:10.1101/2023.08.30.555562</dc:identifier>
<dc:title><![CDATA[The small G-protein Rac1 in the dorsomedial striatum promotes alcohol-dependent structural plasticity and goal-directed learning in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.31.555797v1?rss=1">
<title>
<![CDATA[
Anionic phospholipids determine G protein coupling selectivity at distinct subcellular membrane compartments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.31.555797v1?rss=1</link>
<description><![CDATA[
G protein-coupled receptors (GPCRs) activate different signaling pathways through coupling to heterotrimeric G proteins consisting of four types of G alpha (G) subunits and the beta gamma (yy) subunits. The C-terminal regions of G proteins is thought to determine coupling selectivity. However, there are several reports indicating that some receptors are promiscuous and can couple to multiple G proteins. The precise mechanism promoting promiscuous coupling is not fully understood. Here we show that anionic phospholipids such as phosphoinositide 4,5 bisphosphate (PI(4,5)P2) promote electrostatic interactions between anionic lipid head groups and positively charged amino acids in the transmembrane 4 (TM4) region of beta adrenergic receptors ({beta}ARs) to prime receptor coupling to the cognate Gs and non-cognate Gi proteins. The promiscuous coupling can only occur at the plasma membrane, a membrane compartment enriched in PI(4,5)P2, while receptors only couple to cognate Gs proteins in compartments that are enriched in less negatively charged phosphoinositides such PI4P at the Golgi membranes. We took advantage of the rapamycin dimerization system to rapidly and inducibly deplete PI(4,5)P2 by recruiting a phosphatase to the plasma membrane and found that in the absence of anionic phospholipids, {beta}ARs couple only to their cognate Gs protein. Finally, we found that mutating {beta}ARs PI(4,5)P2 binding motifs or depleting PI(4,5)P2 results in enhanced {beta}ARs-mediated cAMP response. Together, these findings reveal a role for anionic phosphoinositides in regulating GPCR activity at different subcellular compartments.
]]></description>
<dc:creator>Hernandez, E. H.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:creator>Irannejad, R.</dc:creator>
<dc:date>2023-09-01</dc:date>
<dc:identifier>doi:10.1101/2023.08.31.555797</dc:identifier>
<dc:title><![CDATA[Anionic phospholipids determine G protein coupling selectivity at distinct subcellular membrane compartments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.01.555926v1?rss=1">
<title>
<![CDATA[
Dual α-globin and truncated erythropoietin receptor knock-in restores hemoglobin production in α-thalassemia major-derived hematopoietic stem and progenitor cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.01.555926v1?rss=1</link>
<description><![CDATA[
Alpha-thalassemia is an autosomal recessive disease with increasing worldwide prevalence. The molecular basis is due to mutation or deletion of one or more duplicated -globin genes, and disease severity is directly related to the number of allelic copies compromised. The most severe form, -thalassemia major (TM), results from loss of all four copies of -globin and has historically resulted in fatality in utero. However, in utero transfusions now enable survival to birth. Postnatally, patients face challenges similar to {beta}-thalassemia, including severe anemia and erythrotoxicity due to imbalance of {beta}-globin and -globin chains. While curative, hematopoietic stem cell transplantation (HSCT) is limited by donor availability and potential transplant-related complications. Despite progress in genome editing treatments for {beta}-thalassemia, there is no analogous curative option for patients suffering from -thalassemia. To address this, we designed a novel Cas9/AAV6-mediated genome editing strategy that integrates a functional -globin gene into the {beta}-globin locus in TM patient-derived hematopoietic stem and progenitor cells (HSPCs). Incorporation of a truncated erythropoietin receptor transgene into the -globin integration cassette dramatically increased erythropoietic output from edited HSPCs and led to the most robust production of -globin, and consequently normal hemoglobin. By directing edited HSPCs toward increased production of clinically relevant RBCs instead of other divergent cell types, this approach has the potential to mitigate the limitations of traditional HSCT for the hemoglobinopathies, including low genome editing and low engraftment rates. These findings support development of a definitive ex vivo autologous genome editing strategy that may be curative for -thalassemia.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=177 SRC="FIGDIR/small/555926v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Chu, S. N.</dc:creator>
<dc:creator>Soupene, E.</dc:creator>
<dc:creator>Wienert, B.</dc:creator>
<dc:creator>Yin, H.</dc:creator>
<dc:creator>Sharma, D.</dc:creator>
<dc:creator>Jia, K.</dc:creator>
<dc:creator>Homma, S.</dc:creator>
<dc:creator>Hampton, J. P.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:creator>MacKenzie, T. C.</dc:creator>
<dc:creator>Porteus, M. H.</dc:creator>
<dc:creator>Cromer, M. K.</dc:creator>
<dc:date>2023-09-02</dc:date>
<dc:identifier>doi:10.1101/2023.09.01.555926</dc:identifier>
<dc:title><![CDATA[Dual α-globin and truncated erythropoietin receptor knock-in restores hemoglobin production in α-thalassemia major-derived hematopoietic stem and progenitor cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.01.555831v1?rss=1">
<title>
<![CDATA[
In vivo perturb-seq of cancer and immune cells dissects oncologic drivers and therapy response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.01.555831v1?rss=1</link>
<description><![CDATA[
Genetic perturbation screens with single cell readouts have enabled rich phenotyping of gene function and regulatory networks. These approaches have been challenging in vivo, especially in adult disease models such as cancer, which include mixtures of malignant and microenvironment cells. Glioblastoma (GBM) is a fatal cancer, and methods of systematically interrogating gene function and therapeutic targets in vivo, especially in combination with standard of care treatment such as radiotherapy, are lacking. Here, we iteratively develop a multiplex in vivo perturb-seq CRISPRi platform for single cell genetic screens in cancer and tumor microenvironment cells that leverages intracranial convection enhanced delivery (CED) of sgRNA libraries into models of GBM. Our platform enables potent silencing of drivers of in vivo growth and tumor maintenance, as well as genes that sensitize GBM to radiotherapy. We find radiotherapy rewires transcriptional responses to genetic perturbations in an in vivo dependent manner, revealing heterogenous patterns of treatment sensitization or resistance in GBM. Furthermore, we demonstrate targeting of genes that function in the tumor microenvironment, enabling alterations of ligand-receptor interactions between immune/stromal cells following in vivo CRISPRi perturbations. In sum, we demonstrate the utility of multiplexed perturb-seq for in vivo single cell dissection of adult cancer and normal tissue biology across multiple cell types in the context of therapeutic intervention, a platform with potential for broad application.
]]></description>
<dc:creator>Liu, S. J.</dc:creator>
<dc:creator>Pak, J.</dc:creator>
<dc:creator>Zou, C.</dc:creator>
<dc:creator>Casey-Clyde, T.</dc:creator>
<dc:creator>Borah, A.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Seo, K.</dc:creator>
<dc:creator>O'Loughlin, T.</dc:creator>
<dc:creator>Lim, D.</dc:creator>
<dc:creator>Ozawa, T.</dc:creator>
<dc:creator>Berger, M.</dc:creator>
<dc:creator>Weiss, W.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:date>2023-09-03</dc:date>
<dc:identifier>doi:10.1101/2023.09.01.555831</dc:identifier>
<dc:title><![CDATA[In vivo perturb-seq of cancer and immune cells dissects oncologic drivers and therapy response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.02.556060v1?rss=1">
<title>
<![CDATA[
Reward expectation selectively boosts the firing of accumbens D1+ neurons during motivated approach. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.02.556060v1?rss=1</link>
<description><![CDATA[
The nucleus accumbens (NAc) helps govern motivation to pursue rewards. Two distinct sets of NAc projection neurons-expressing dopamine D1 versus D2 receptors-are thought to promote and suppress motivated behaviors respectively. However, support for this conceptual framework is limited: in particular the spiking patterns of these distinct cell types during motivated behavior have been largely unknown. We monitored identified D1+ and D2+ neurons in the NAc Core, as unrestrained rats performed an operant task in which motivation to initiate work tracks recent reward rate. D1+ neurons preferentially increased firing as rats initiated trials, and fired more when reward expectation was higher. By contrast, D2+ cells preferentially increased firing later in the trial especially in response to reward delivery - a finding not anticipated from current theoretical models. Our results provide new evidence for the specific contribution of NAc D1+ cells to self-initiated approach behavior, and will spur updated models of how we learn from rewards.
]]></description>
<dc:creator>Faust, T.</dc:creator>
<dc:creator>Mohebi, A.</dc:creator>
<dc:creator>Berke, J. D.</dc:creator>
<dc:date>2023-09-03</dc:date>
<dc:identifier>doi:10.1101/2023.09.02.556060</dc:identifier>
<dc:title><![CDATA[Reward expectation selectively boosts the firing of accumbens D1+ neurons during motivated approach.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.04.555754v1?rss=1">
<title>
<![CDATA[
Defining RNA oligonucleotides that reverse deleterious phase transitions of RNA-binding proteins with prion-like domains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.04.555754v1?rss=1</link>
<description><![CDATA[
RNA-binding proteins with prion-like domains, such as FUS and TDP-43, condense into functional liquids, which can transform into pathological fibrils that underpin fatal neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS)/frontotemporal dementia (FTD). Here, we define short RNAs (24-48 nucleotides) that prevent FUS fibrillization by promoting liquid phases, and distinct short RNAs that prevent and, remarkably, reverse FUS condensation and fibrillization. These activities require interactions with multiple RNA-binding domains of FUS and are encoded by RNA sequence, length, and structure. Importantly, we define a short RNA that dissolves aberrant cytoplasmic FUS condensates, restores nuclear FUS, and mitigates FUS proteotoxicity in optogenetic models and human motor neurons. Another short RNA dissolves aberrant cytoplasmic TDP-43 condensates, restores nuclear TDP-43, and mitigates TDP-43 proteotoxicity. Since short RNAs can be effectively delivered to the human brain, these oligonucleotides could have therapeutic utility for ALS/FTD and related disorders.
]]></description>
<dc:creator>Guo, L.</dc:creator>
<dc:creator>Mann, J. R.</dc:creator>
<dc:creator>Mauna, J. C.</dc:creator>
<dc:creator>Copley, K. E.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Rubien, J. D.</dc:creator>
<dc:creator>Odeh, H. M.</dc:creator>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Lee, B. L.</dc:creator>
<dc:creator>Ganser, L.</dc:creator>
<dc:creator>Robinson, E.</dc:creator>
<dc:creator>Kim, K. M.</dc:creator>
<dc:creator>Murthy, A. C.</dc:creator>
<dc:creator>Paul, T.</dc:creator>
<dc:creator>Portz, B.</dc:creator>
<dc:creator>Gleixner, A. M.</dc:creator>
<dc:creator>Diaz, Z.</dc:creator>
<dc:creator>Carey, J. L.</dc:creator>
<dc:creator>Smirnov, A.</dc:creator>
<dc:creator>Padilla, G.</dc:creator>
<dc:creator>Lavorando, E.</dc:creator>
<dc:creator>Espy, C.</dc:creator>
<dc:creator>Shang, Y.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Chesi, A.</dc:creator>
<dc:creator>Fawzi, N. L.</dc:creator>
<dc:creator>Myong, S.</dc:creator>
<dc:creator>Donnelly, C. J.</dc:creator>
<dc:creator>Shorter, J.</dc:creator>
<dc:date>2023-09-04</dc:date>
<dc:identifier>doi:10.1101/2023.09.04.555754</dc:identifier>
<dc:title><![CDATA[Defining RNA oligonucleotides that reverse deleterious phase transitions of RNA-binding proteins with prion-like domains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.04.556039v1?rss=1">
<title>
<![CDATA[
RNA binding proteins and glycoRNAs form domains on the cell surface for cell penetrating peptide entry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.04.556039v1?rss=1</link>
<description><![CDATA[
The composition and organization of the cell surface determine how cells interact with their environment. Traditionally, glycosylated transmembrane proteins were thought to be the major constituents of the external surface of the plasma membrane. Here, we provide evidence that a group of RNA binding proteins (RBPs) are present on the surface of living cells. These cell surface RBPs (csRBPs) precisely organize into well-defined nanoclusters that are enriched for multiple RBPs, glycoRNAs, and their clustering can be disrupted by extracellular RNase addition. These glycoRNA-csRBP clusters further serve as sites of cell surface interaction for the cell penetrating peptide TAT. Removal of RNA from the cell surface, or loss of RNA binding activity by TAT, causes defects in TAT cell internalization. Together, we provide evidence of an expanded view of the cell surface by positioning glycoRNA-csRBP clusters as a regulator of communication between cells and the extracellular environment.
]]></description>
<dc:creator>Perr, J.</dc:creator>
<dc:creator>Langen, A.</dc:creator>
<dc:creator>Almahayni, K.</dc:creator>
<dc:creator>Nestola, G.</dc:creator>
<dc:creator>Chai, P.</dc:creator>
<dc:creator>Lebedenko, C.</dc:creator>
<dc:creator>Volk, R.</dc:creator>
<dc:creator>Caldwell, R.</dc:creator>
<dc:creator>Spiekermann, M.</dc:creator>
<dc:creator>Hemberger, H.</dc:creator>
<dc:creator>Bisaria, N.</dc:creator>
<dc:creator>Tzelepis, K.</dc:creator>
<dc:creator>Calo, E.</dc:creator>
<dc:creator>Mockl, L.</dc:creator>
<dc:creator>Zaro, B.</dc:creator>
<dc:creator>Flynn, R. A.</dc:creator>
<dc:date>2023-09-05</dc:date>
<dc:identifier>doi:10.1101/2023.09.04.556039</dc:identifier>
<dc:title><![CDATA[RNA binding proteins and glycoRNAs form domains on the cell surface for cell penetrating peptide entry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.05.556133v1?rss=1">
<title>
<![CDATA[
Characterizing tissue structures from spatial omics with spatial cellular graph partition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.05.556133v1?rss=1</link>
<description><![CDATA[
Spatial transcriptomic and proteomic measurements enable high-dimensional characterization of tissues. However, understanding organizations of cells at different spatial scales and extracting tissue structures of interest remain challenging tasks that require extensive human annotations. To address this need for consistent identification of tissue structures, in this work, we present a novel annotation method Spatial Cellular Graph Partitioning (SCGP) that allows unsupervised identification of tissue structures that reflect the anatomical and functional units of human tissues. We further present a reference-query extension pipeline SCGP-Extension that enables the generalization of existing reference tissue structures to previously unseen samples. Our experiments demonstrate reliable and robust partitionings of both spatial transcriptomics and proteomics datasets encompassing different tissue types and profiling techniques. Downstream analysis on SCGP-identified tissue structures reveals disease-relevant insights regarding diabetic kidney disease and skin disorder, underscoring its potential in facilitating spatial analysis and driving new discoveries.
]]></description>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Kondo, A.</dc:creator>
<dc:creator>McGrady, M.</dc:creator>
<dc:creator>Baker, E. A. G.</dc:creator>
<dc:creator>Wu, E.</dc:creator>
<dc:creator>Rahim, M. K.</dc:creator>
<dc:creator>Bracey, N. A.</dc:creator>
<dc:creator>Charu, V.</dc:creator>
<dc:creator>Cho, R. J.</dc:creator>
<dc:creator>Cheng, J. B.</dc:creator>
<dc:creator>Afkarian, M.</dc:creator>
<dc:creator>Zou, J.</dc:creator>
<dc:creator>Mayer, A. T.</dc:creator>
<dc:creator>Trevino, A. E.</dc:creator>
<dc:date>2023-09-05</dc:date>
<dc:identifier>doi:10.1101/2023.09.05.556133</dc:identifier>
<dc:title><![CDATA[Characterizing tissue structures from spatial omics with spatial cellular graph partition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.06.556529v1?rss=1">
<title>
<![CDATA[
Causal Genetic Loci for a Motivated Behavior Spectrum Harbor Psychiatric Risk Genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.06.556529v1?rss=1</link>
<description><![CDATA[
Behavioral diversity is critical for population fitness. Individual differences in risk-taking are observed across species, but underlying genetic mechanisms and conservation are largely unknown. We examined dark avoidance in larval zebrafish, a motivated behavior reflecting an approach-avoidance conflict. Brain-wide calcium imaging revealed significant neural activity differences between approach-inclined versus avoidance-inclined individuals. We used a population of [~]6,000 to perform the first genome-wide association study (GWAS) in zebrafish, which identified 34 genomic regions harboring many genes that are involved in synaptic transmission and human psychiatric diseases. We used CRISPR to study several causal genes: serotonin receptor-1b (htr1b), nitric oxide synthase-1 (nos1), and stress-induced phosphoprotein-1 (stip1). We further identified 52 conserved elements containing 66 GWAS significant variants. One encoded an exonic regulatory element that influenced tissue-specific nos1 expression. Together, these findings reveal new genetic loci and establish a powerful, scalable animal system to probe mechanisms underlying motivation, a critical dimension of psychiatric diseases.
]]></description>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Casanave, R.</dc:creator>
<dc:creator>Chitre, A. S.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Nguyen, K.-M.</dc:creator>
<dc:creator>Blake, C.</dc:creator>
<dc:creator>Wagle, M.</dc:creator>
<dc:creator>Cheng, R.</dc:creator>
<dc:creator>Polesskaya, O.</dc:creator>
<dc:creator>Palmer, A. A.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:date>2023-09-06</dc:date>
<dc:identifier>doi:10.1101/2023.09.06.556529</dc:identifier>
<dc:title><![CDATA[Causal Genetic Loci for a Motivated Behavior Spectrum Harbor Psychiatric Risk Genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.07.556260v1?rss=1">
<title>
<![CDATA[
A soybean rust effector protease suppresses host immunity and cleaves a 3-deoxy-7-phosphoheptulonate synthase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.07.556260v1?rss=1</link>
<description><![CDATA[
The devastating soybean rust (SBR) pathogen, Phakopsora pachyrhizi, encodes many secreted proteins, but only two have been functionally characterized for their roles in rust virulence. Here, we demonstrate that transient expression of P. pachyrhizi effector candidate 15 (PpEC15), an aspartic protease, leads to enhanced bacterial growth in planta, suppression of callose deposition, reduced expression of plant defense-related marker genes and suppression of pathogen-associated molecular pattern (PAMP)-induced reactive oxygen species (ROS). Stable expression of PpEC15 in soybean suppresses PAMP-induced ROS production and enhances bacterial growth, indicating that, collectively, PpEC15 suppresses host and non-host innate immune responses. Yeast-two-hybrid and proximity labeling identified putative PpEC15 interacting partners including a peptide-chain release factor (PCRF), a NAC83 (NAM, ATAF, and CUC) transcription factor, and a DAHP (3-deoxy-7-phosphoheptulonate) synthase. We further show that PpEC15 can cleave DAHP but does not cleave PCRF or NAC83. Virus-induced gene silencing of NAC83, PCRF and DAHP altered PAMP-induced ROS production and salicylic acid production, indicating that these proteins may be involved in immune signaling. Collectively, our data show that PpEC15 is conserved across P. pachyrhizi isolates and other economically important rust species and is involved in the suppression of plant basal defense responses. Understanding the role of PpEC15 in P. pachyrhizi virulence will provide a foundation for designing targeted intervention strategies to generate rust-resistant crops.
]]></description>
<dc:creator>Chicowski, A. S.</dc:creator>
<dc:creator>Qi, M.</dc:creator>
<dc:creator>Variz, H.</dc:creator>
<dc:creator>Bredow, M.</dc:creator>
<dc:creator>Montes-Serey, C.</dc:creator>
<dc:creator>Caiazza, F.</dc:creator>
<dc:creator>Dong, H.</dc:creator>
<dc:creator>Margets, A. C.</dc:creator>
<dc:creator>Mejias, J.</dc:creator>
<dc:creator>Walley, J.</dc:creator>
<dc:creator>Craik, C.</dc:creator>
<dc:creator>Pedley, K. F.</dc:creator>
<dc:creator>Aung, K.</dc:creator>
<dc:creator>Innes, R. W.</dc:creator>
<dc:creator>Whitham, S.</dc:creator>
<dc:date>2023-09-08</dc:date>
<dc:identifier>doi:10.1101/2023.09.07.556260</dc:identifier>
<dc:title><![CDATA[A soybean rust effector protease suppresses host immunity and cleaves a 3-deoxy-7-phosphoheptulonate synthase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.07.556726v1?rss=1">
<title>
<![CDATA[
Endogenous Opioid Signaling Regulates Proliferation of Spinal Cord Ependymal Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.07.556726v1?rss=1</link>
<description><![CDATA[
After injury, mammalian spinal cords develop scars to seal off the damaged area and prevent further injury. However, excessive scarring can hinder neural regeneration and functional recovery (1, 2). These competing actions underscore the importance of developing therapeutic strategies to dynamically modulate the extent of scar formation. Previous research on scar formation has primarily focused on the role of astrocytes, but recent evidence suggests that ependymal cells also participate. Ependymal cells normally form the epithelial layer encasing the central canal, but they undergo massive proliferation and differentiation into astroglia following certain types of injury, becoming a core component of scars (3-7). However, the mechanisms regulating ependymal proliferation in vivo in both healthy and injured conditions remain unclear. Here, we uncover an intercellular kappa ({kappa}) opioid signaling pathway that controls endogenous ependymal proliferation. Specifically, we detect expression of the {kappa} opioid receptor, OPRK1, in a functionally under-characterized cell type called cerebrospinal fluid-contacting neurons (CSF-cNs). We also discover a neighboring cell population that express the cognate ligand, prodynorphin (PDYN). Importantly, OPRK1 activation excites CSF-cNs, and systemic administration of a {kappa} antagonist enhances ependymal proliferation in uninjured spinal cords in a CSF-cN-dependent manner. Moreover, injecting a {kappa} agonist reduces the proliferation induced by dorsal hemisection. Altogether, our data suggest a regulatory mechanism whereby PDYN+ cells tonically release {kappa} opioids to stimulate CSF-cNs, which in turn suppress ependymal proliferation. This endogenous pathway provides a mechanistic basis for the potential use of {kappa} opiates in modulating scar formation and treating spinal cord injuries.
]]></description>
<dc:creator>Yue, W. W. S.</dc:creator>
<dc:creator>Touhara, K. K.</dc:creator>
<dc:creator>Toma, K.</dc:creator>
<dc:creator>Duan, X.</dc:creator>
<dc:creator>Julius, D.</dc:creator>
<dc:date>2023-09-08</dc:date>
<dc:identifier>doi:10.1101/2023.09.07.556726</dc:identifier>
<dc:title><![CDATA[Endogenous Opioid Signaling Regulates Proliferation of Spinal Cord Ependymal Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.07.556760v1?rss=1">
<title>
<![CDATA[
Clonal stochasticity in early NK cell response to mouse cytomegalovirus is generated by mature subsets of varying proliferative ability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.07.556760v1?rss=1</link>
<description><![CDATA[
Natural killer (NK) cells are classically defined as innate immune cells, but experiments show that mouse cytomegalovirus (MCMV) infection in C57BL/6 mice can cause NK cells to undergo antigen-specific proliferation and memory formation, similar to adaptive CD8+ T cells. One shared behavior between CD8+ T cells and NK cells is clonal expansion, where a single stimulated cell proliferates rapidly to form a diverse population of cells. For example, clones derived from single cells are most abundant during expansion when they are primarily CD27-for NK cells and CD62L-for T cells, phenotypes derived from precursor CD27+ and CD62L+ cells, respectively. Here we determined the mechanistic rules involving proliferation, cell death, and differentiation of endogenous and adoptively transferred NK cells in the expansion phase of the response to MCMV infection. We found that the interplay between cell proliferation and cell death of mature CD27-NK cells and a highly proliferative CD27-Ly6C-mature subtype and intrinsic stochastic fluctuations in these processes play key roles in regulating the heterogeneity and population of the NK cell subtypes. Furthermore, we estimate rates for maturation of endogenous NK cells in homeostasis and in MCMV infection and found that only NK cell growth rates, and not differentiation rates, are appreciably increased by MCMV. Taken together, these results quantify the differences between the kinetics of NK cell antigen-specific expansion from that of CD8+ T cells and unique mechanisms that give rise to the observed heterogeneity in NK cell clones generated from single NK cells in the expansion phase.
]]></description>
<dc:creator>Wethington, D.</dc:creator>
<dc:creator>Potempa, M.</dc:creator>
<dc:creator>Giuliani, G.</dc:creator>
<dc:creator>Aguilar, O. A.</dc:creator>
<dc:creator>Grassmann, S.</dc:creator>
<dc:creator>Stewart, W.</dc:creator>
<dc:creator>Adams, N. M.</dc:creator>
<dc:creator>Sun, J. C.</dc:creator>
<dc:creator>Lanier, L. L.</dc:creator>
<dc:creator>Das, J.</dc:creator>
<dc:date>2023-09-08</dc:date>
<dc:identifier>doi:10.1101/2023.09.07.556760</dc:identifier>
<dc:title><![CDATA[Clonal stochasticity in early NK cell response to mouse cytomegalovirus is generated by mature subsets of varying proliferative ability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.07.556550v1?rss=1">
<title>
<![CDATA[
TRPM2-CaMKII signaling drives excessive GABAergic synaptic inhibition following ischemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.07.556550v1?rss=1</link>
<description><![CDATA[
Following an ischemic insult to the brain, there is an acute loss of GABAergic inhibitory synapses and an increase in excitatory/ inhibitory (E/I) imbalance that drives neuronal hyperexcitability. It is unknown whether this E/I imbalance persists at delayed timepoints and contributes to chronic impairments in memory and long-term potentiation (LTP) in the hippocampus following ischemic brain injury. Here, we reveal a shift to reduced E/I ratio in hippocampal CA1 neurons via a persistent increase in postsynaptic GABAA receptor mediated inhibitory responses and clustering days after a global ischemic insult. This enhancement of postsynaptic inhibitory function and clustering required activation of the Ca2+-permeable TRPM2 ion channel and the Ca2+-dependent kinase, CaMKII. Thus, we propose a mechanism in which acute downregulation of GABAA receptors is followed by a strengthening of inhibitory synapses at delayed periods after ischemia. Targeting this mechanism has therapeutic potential to recover hippocampal plasticity and cognitive function post-ischemia.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=123 SRC="FIGDIR/small/556550v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Burch, A. M.</dc:creator>
<dc:creator>Garcia, J. D.</dc:creator>
<dc:creator>O'Leary, H.</dc:creator>
<dc:creator>Haas, A.</dc:creator>
<dc:creator>Orfila, J. E.</dc:creator>
<dc:creator>Tiemeier, E.</dc:creator>
<dc:creator>Chalmers, N.</dc:creator>
<dc:creator>Smith, K. R.</dc:creator>
<dc:creator>Quillinan, N.</dc:creator>
<dc:creator>Herson, P. S.</dc:creator>
<dc:date>2023-09-09</dc:date>
<dc:identifier>doi:10.1101/2023.09.07.556550</dc:identifier>
<dc:title><![CDATA[TRPM2-CaMKII signaling drives excessive GABAergic synaptic inhibition following ischemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.08.556815v1?rss=1">
<title>
<![CDATA[
Probing the Mechanisms of Global Brain Reconfiguration after Local Manipulations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.08.556815v1?rss=1</link>
<description><![CDATA[
Understanding how localized brain interventions translate into whole-brain dynamics is crucial for deciphering neural function and tailoring therapeutic strategies. Combining mouse experimental datasets of focal interventions (thalamic lesion and chemogenetic silencing of cortical hubs), we demonstrate both local and global effects. Using whole-brain simulations of experimental data, we not only confirm the distributed nature of local manipulations but also offer mechanistic insights into these processes. Our simulations predict specific alterations in firing rates and spectral characteristics across specific brain networks, leading to structured changes in functional connectivity patterns. Some of these predictions have been empirically validated. Notably, the affected brain subnetworks--and their resultant  signatures of change--are contingent on the original intervention site, suggesting a method to accurately localize the source of alteration. Our results provide a general framework for interpreting localized intervention effects, offering insights that could refine clinical interventions for focal brain disorders by enabling targeted circuit-level neuromodulation strategies.
]]></description>
<dc:creator>Rabuffo, G.</dc:creator>
<dc:creator>Lokossou, H.-A.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Ziaee-Mehr, A.</dc:creator>
<dc:creator>Hashemi, M.</dc:creator>
<dc:creator>Quilichini, P. P.</dc:creator>
<dc:creator>Ghestem, A.</dc:creator>
<dc:creator>Arab, O.</dc:creator>
<dc:creator>Esclapez, M.</dc:creator>
<dc:creator>Verma, P.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Gozzi, A.</dc:creator>
<dc:creator>Sorrentino, P.</dc:creator>
<dc:creator>Chuang, K.-H.</dc:creator>
<dc:creator>Perles-Barbacaru, T.-A.</dc:creator>
<dc:creator>Viola, A.</dc:creator>
<dc:creator>Jirsa, V. K.</dc:creator>
<dc:creator>Bernard, C.</dc:creator>
<dc:date>2023-09-09</dc:date>
<dc:identifier>doi:10.1101/2023.09.08.556815</dc:identifier>
<dc:title><![CDATA[Probing the Mechanisms of Global Brain Reconfiguration after Local Manipulations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.08.556847v1?rss=1">
<title>
<![CDATA[
A General Method to Accurately Count Molecular Complexes and Determine the Degree of Labelling in Cells Using Protein Tags 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.08.556847v1?rss=1</link>
<description><![CDATA[
Determining the label to target ratio, also known as degree of labelling (DOL), is crucial for quantitative fluorescence microscopy and a high DOL with minimal unspecific labelling is beneficial for fluorescence microscopy in general. Yet, robust, versatile, and easy-to-use tools for measuring cell-specific labelling efficiencies are not available. This study presents a novel DOL determination technique named Protein-tag DOL (ProDOL), which enables fast DOL measurements and optimisation of protein-tag labelling. With ProDOL various factors affecting labelling efficiency, including substrate type, incubation time, and concentration, as well as sample fixation and cell type can be easily assessed. We applied ProDOL to investigate how HIV-1 pathogenesis factor Nef modulates CD4 T cell activation measuring total and activated copy numbers of the adaptor protein SLP-76 in signalling microclusters. ProDOL proved to be a versatile and robust tool for labelling calibration, enabling determination of labelling efficiencies, optimisation of strategies, and quantification of protein stoichiometry.
]]></description>
<dc:creator>Tashev, S. A.</dc:creator>
<dc:creator>Euchner, J.</dc:creator>
<dc:creator>Yserentant, K.</dc:creator>
<dc:creator>Hänselmann, S.</dc:creator>
<dc:creator>Hild, F.</dc:creator>
<dc:creator>Chmielewicz, W.</dc:creator>
<dc:creator>Hummert, J.</dc:creator>
<dc:creator>Schwörer, F.</dc:creator>
<dc:creator>Tsopoulidis, N.</dc:creator>
<dc:creator>Germer, S.</dc:creator>
<dc:creator>Sassmannshausen, Z.</dc:creator>
<dc:creator>Fackler, O. T.</dc:creator>
<dc:creator>Klingmüller, U.</dc:creator>
<dc:creator>Herten, D.-P.</dc:creator>
<dc:date>2023-09-09</dc:date>
<dc:identifier>doi:10.1101/2023.09.08.556847</dc:identifier>
<dc:title><![CDATA[A General Method to Accurately Count Molecular Complexes and Determine the Degree of Labelling in Cells Using Protein Tags]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.08.556878v1?rss=1">
<title>
<![CDATA[
CCR3 inhibition suppresses inflammation-driven recruitment of peripheral immune cells to the eye 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.08.556878v1?rss=1</link>
<description><![CDATA[
C-C chemokine receptor type 3 (CCR3) has been linked with age-related macular degeneration (AMD) pathologies. Specifically, its function as an immune modulator in AMD remains unclear. To address this question, we investigated the impact of CCR3 inhibition on inflammation, a key driver of AMD pathologies, by assessing inflammatory cytokines and infiltrating immune cells in two models of ocular inflammation.

Mice were orally dosed twice a day with AKST4290, a CCR3 small molecular inhibitor, in the sodium iodate (NaIO3) and myelin oligodendrocyte glycoprotein (MOG) models. A combination of autoradiography and analytical chemistry techniques were used to assess drug concentration and distribution. Bead-based multiplexing technology was used to determine cytokine concentrations, and flow cytometry and immunohistochemistry was used to ascertain ocular immune-cell composition.

CCR3 expression was detected in the retinal pigment epithelium (RPE)/choroid complex where AKST4290 was found to preferentially accumulate at sustained levels. In the NaIO3 model, inhibition of CCR3 with AKST4290 significantly decreased both the concentration of specific chemokines and the number of multiple populations of infiltrating peripheral immune cell. Furthermore, effects of CCR3 inhibition on immune cell infiltration were confirmed in the MOG model.

These data demonstrate that CCR3 inhibition strongly modulates local inflammation by impacting both cytokine concentrations and immune cell composition in ocular diseases.

Moreover, these findings together with the known role of CCR3 in promoting pathologic angiogenesis implicate a pleiotropic role for CCR3 in AMD.
]]></description>
<dc:creator>Lopez, Y.</dc:creator>
<dc:creator>Caryotakis, S.</dc:creator>
<dc:creator>Raina, S.</dc:creator>
<dc:creator>Rege, S. V.</dc:creator>
<dc:creator>Harish, R.</dc:creator>
<dc:creator>Ray, R.</dc:creator>
<dc:creator>Kosti, I.</dc:creator>
<dc:creator>Teichert, A.</dc:creator>
<dc:creator>Minami, S. S.</dc:creator>
<dc:creator>Dhande, O.</dc:creator>
<dc:date>2023-09-09</dc:date>
<dc:identifier>doi:10.1101/2023.09.08.556878</dc:identifier>
<dc:title><![CDATA[CCR3 inhibition suppresses inflammation-driven recruitment of peripheral immune cells to the eye]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.06.556606v1?rss=1">
<title>
<![CDATA[
Reciprocal inflammatory signals establish profibrotic cross-feeding metabolism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.06.556606v1?rss=1</link>
<description><![CDATA[
Idiopathic pulmonary fibrosis (IPF) is a disease of progressive lung remodeling and collagen deposition that leads to respiratory failure. Myeloid cells are abundant in IPF lung and in murine lung fibrosis, but their functional effects are incompletely understood. Using mouse and human lung models, we show that ornithine produced by myeloid cells expressing Arg1 serves as a substrate for proline and collagen synthesis by lung fibroblasts. The predominant Arg1-expressing myeloid cells in mouse lung were macrophages, but in IPF lung, high-dimensional imaging revealed ARG1 to be expressed mainly in neutrophils. Arg1 inhibition suppressed both ornithine levels and collagen expression in cultured, precision-cut IPF lung slices and in murine lung fibrosis. These results were confirmed in macrophage-specific Arg1 KO mice. Furthermore, we find that this pathway is regulated by cell-to-cell crosstalk, starting with purinergic signaling: Fibroblast eATP receptor P2rx4 was necessary for fibroblast IL-6 expression, which in turn was necessary for Arg1 expression by myeloid cells. Taken together, our findings define an immune-mesenchymal circuit that governs profibrotic metabolism in lung fibrosis.
]]></description>
<dc:creator>Yadav, P.</dc:creator>
<dc:creator>Gomez Ortega, J.</dc:creator>
<dc:creator>Chang, K.-C.</dc:creator>
<dc:creator>Pan, S.</dc:creator>
<dc:creator>Biswas, N.</dc:creator>
<dc:creator>Nilsson, J.</dc:creator>
<dc:creator>Yin, X.</dc:creator>
<dc:creator>Bhattacharyya, A.</dc:creator>
<dc:creator>Boostanpour, K.</dc:creator>
<dc:creator>Tsukui, T.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Maishan, M.</dc:creator>
<dc:creator>Taenaka, H.</dc:creator>
<dc:creator>Matthay, M.</dc:creator>
<dc:creator>Muramatsu, R.</dc:creator>
<dc:creator>Naser, M.</dc:creator>
<dc:creator>Molofsky, A. B.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:creator>Abate, A. R.</dc:creator>
<dc:creator>Tharp, K.</dc:creator>
<dc:creator>Bhattacharya, M.</dc:creator>
<dc:date>2023-09-10</dc:date>
<dc:identifier>doi:10.1101/2023.09.06.556606</dc:identifier>
<dc:title><![CDATA[Reciprocal inflammatory signals establish profibrotic cross-feeding metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.11.557250v1?rss=1">
<title>
<![CDATA[
Biological and Genetic Determinants of Red Blood Cell Glycolysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.11.557250v1?rss=1</link>
<description><![CDATA[
Mature red blood cells (RBCs) lack mitochondria, and thus exclusively rely on glycolysis to generate adenosine triphosphate (ATP) during aging in vivo or storage in the blood bank. Here we leveraged 13,029 volunteers from the Recipient Epidemiology and Donor Evaluation Study to identify an association between end-of-storage levels of glycolytic metabolites and donor age, sex, and ancestry-specific genetic polymorphisms in regions encoding phosphofructokinase 1, platelet (detected in mature RBCs), hexokinase 1, ADP-ribosyl cyclase 1 and 2 (CD38/BST1). Gene-metabolite associations were validated in fresh and stored RBCs from 525 Diversity Outbred mice, and via multi-omics characterization of 1,929 samples from 643 human RBC units during storage. ATP and hypoxanthine levels - and the genetic traits linked to them - were associated with hemolysis in vitro and in vivo, both in healthy autologous transfusion recipients and in 5,816 critically ill patients receiving heterologous transfusions, suggesting their potential as markers to improve transfusion outcomes.

eTOC and Highlights

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=199 SRC="FIGDIR/small/557250v4_ufig1.gif" ALT="Figure 1">
View larger version (87K):
org.highwire.dtl.DTLVardef@1a556b7org.highwire.dtl.DTLVardef@1e02b13org.highwire.dtl.DTLVardef@2bfab1org.highwire.dtl.DTLVardef@15787e3_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LIBlood donor age and sex affect glycolysis in stored RBCs from 13,029 volunteers;
C_LIO_LIAncestry, genetic polymorphisms in PFKP, HK1, CD38/BST1 influence RBC glycolysis;
C_LIO_LIModeled PFKP effects relate to preventing loss of the total AXP pool in stored RBCs;
C_LIO_LIATP and hypoxanthine are biomarkers of hemolysis in vitro and in vivo.
C_LI
]]></description>
<dc:creator>Nemkov, T.</dc:creator>
<dc:creator>Stephenson, D.</dc:creator>
<dc:creator>Earley, E. J.</dc:creator>
<dc:creator>Keele, G. R.</dc:creator>
<dc:creator>Hay, A.</dc:creator>
<dc:creator>Erickson, C.</dc:creator>
<dc:creator>Dzieciatkowska, M.</dc:creator>
<dc:creator>Key, A. M.</dc:creator>
<dc:creator>Moore, A.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Kleinman, S.</dc:creator>
<dc:creator>Spitalnik, S. L.</dc:creator>
<dc:creator>Hod, E. A.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>Churchill, G. A.</dc:creator>
<dc:creator>Roubinian, N.</dc:creator>
<dc:creator>Norris, P. J.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>Zimring, J. C.</dc:creator>
<dc:creator>Page, G. P.</dc:creator>
<dc:creator>D'Alessandro, A.</dc:creator>
<dc:date>2023-09-11</dc:date>
<dc:identifier>doi:10.1101/2023.09.11.557250</dc:identifier>
<dc:title><![CDATA[Biological and Genetic Determinants of Red Blood Cell Glycolysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.09.557002v1?rss=1">
<title>
<![CDATA[
Structure-based discovery of CFTR potentiators and inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.09.557002v1?rss=1</link>
<description><![CDATA[
The cystic fibrosis transmembrane conductance regulator (CFTR) is a crucial ion channel whose loss of function leads to cystic fibrosis, while its hyperactivation leads to secretory diarrhea. Small molecules that improve CFTR folding (correctors) or function (potentiators) are clinically available. However, the only potentiator, ivacaftor, has suboptimal pharmacokinetics and inhibitors have yet to be clinically developed. Here we combine molecular docking, electrophysiology, cryo-EM, and medicinal chemistry to identify novel CFTR modulators. We docked [~]155 million molecules into the potentiator site on CFTR, synthesized 53 test ligands, and used structure-based optimization to identify candidate modulators. This approach uncovered novel mid-nanomolar potentiators as well as inhibitors that bind to the same allosteric site. These molecules represent potential leads for the development of more effective drugs for cystic fibrosis and secretory diarrhea, demonstrating the feasibility of large-scale docking for ion channel drug discovery.
]]></description>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>Kaplan, A. L.</dc:creator>
<dc:creator>Levring, J.</dc:creator>
<dc:creator>Einsiedel, J.</dc:creator>
<dc:creator>Tiedt, S.</dc:creator>
<dc:creator>Distler, K.</dc:creator>
<dc:creator>Omattage, N.</dc:creator>
<dc:creator>Kondratov, I. S.</dc:creator>
<dc:creator>Moroz, Y. S.</dc:creator>
<dc:creator>Pietz, H. L.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Gmeiner, P.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:date>2023-09-12</dc:date>
<dc:identifier>doi:10.1101/2023.09.09.557002</dc:identifier>
<dc:title><![CDATA[Structure-based discovery of CFTR potentiators and inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.11.556924v1?rss=1">
<title>
<![CDATA[
Rab35 is required for embryonic development and kidney and ureter homeostasis through regulation of epithelial cell junctions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.11.556924v1?rss=1</link>
<description><![CDATA[
Structured AbstractO_ST_ABSBackgroundC_ST_ABSRab35 is a member of a GTPase family of endocytic trafficking proteins. Studies in cell lines have indicated that Rab35 participates in cell adhesion, polarity, cytokinesis, and primary cilia length and composition. Additionally, sea urchin Rab35 regulates actin organization and is required for gastrulation. In mice, loss of Rab35 in the CNS disrupts hippocampal development and neuronal organization. Outside of the CNS, the functions of mammalian Rab35 in vivo are unknown.

MethodsWe generated and analyzed the consequences of both congenital and conditional null Rab35 mutations in mice. Using a LacZ reporter allele, we assessed Rab35 expression during development and postnatally. We assessed Rab35 loss in the kidney and ureter using histology, immunofluorescence microscopy, and western blotting.

ResultsCongenital Rab35 loss of function caused embryonic lethality: homozygous mutants arrested at E7.5 with cardiac edema. Conditional loss of Rab35, either during gestation or postnatally, caused hydronephrosis. The kidney and ureter phenotype were associated with disrupted actin cytoskeletal architecture, altered Arf6 epithelial polarity, reduced adherens junctions, loss of tight junction formation, defects in EGFR expression and localization, disrupted cell differentiation, and shortened primary cilia.

ConclusionRab35 is essential for mammalian development and the maintenance of kidney and ureter architecture. Loss of Rab35 leads to non-obstructive hydronephrosis, making the Rab35 mutant mouse a novel mammalian model to study mechanisms underlying this disease.

Significance StatementHydronephrosis, distention of the renal calyces and pelvis, affects 1 in 100 infants. Most cases of hydronephrosis are associated with obstruction. Non-obstructive hydronephrosis is typically associated with impaired ureter development, and requires surgical intervention. Here, we describe a mouse model of non-obstructive hydronephrosis caused by mutations in Rab35. Hydronephrosis in Rab35 mutants is associated with the inability to maintain epithelial cell junctions, defects in EGFR expression, and altered urothelium and smooth muscle integrity of the ureter. The Rab35 mutant mouse is a novel model to study mechanisms and treatment strategies for non-obstructive hydronephrosis.
]]></description>
<dc:creator>Clearman, K. R.</dc:creator>
<dc:creator>Timpratoom, N.</dc:creator>
<dc:creator>Patel, D.</dc:creator>
<dc:creator>Rains, A. B.</dc:creator>
<dc:creator>Haycraft, C. J. R.</dc:creator>
<dc:creator>Croyle, M. J.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:creator>Yoder, B. K.</dc:creator>
<dc:date>2023-09-12</dc:date>
<dc:identifier>doi:10.1101/2023.09.11.556924</dc:identifier>
<dc:title><![CDATA[Rab35 is required for embryonic development and kidney and ureter homeostasis through regulation of epithelial cell junctions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.11.557005v1?rss=1">
<title>
<![CDATA[
Disease related changes in ATAC-Seq of more than 450 iPSC-derived motor neuron lines from ALS patients and controls 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.11.557005v1?rss=1</link>
<description><![CDATA[
Amyotrophic Lateral Sclerosis (ALS), like many other neurodegenerative diseases, is highly heritable, but with only a small fraction of cases explained by monogenic disease alleles. To better understand sporadic ALS, we report epigenomic profiles, as measured by ATAC-seq, of motor neuron cultures derived from a diverse group of 380 ALS patients and 80 healthy controls. We find that chromatin accessibility is heavily influenced by sex, the iPSC cell type of origin, ancestry, and the inherent variance arising from sequencing. Once these covariates are corrected for, we are able to identify robust ALS-specific signals in the data. Additionally, we find that the ATAC-seq data is able to predict ALS disease progression rates with similar accuracy to methods based on biomarkers and clinical status. These results suggest that iPSC-derived motor neurons recapitulate important disease-relevant epigenomic changes.
]]></description>
<dc:creator>Tsitkov, S.</dc:creator>
<dc:creator>Valentine, K.</dc:creator>
<dc:creator>Kozareva, V.</dc:creator>
<dc:creator>Donde, A.</dc:creator>
<dc:creator>Frank, A.</dc:creator>
<dc:creator>Lei, S.</dc:creator>
<dc:creator>Answer ALS Consortium,</dc:creator>
<dc:creator>Van Eyk, J.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:creator>Rothstein, J.</dc:creator>
<dc:creator>Thompson, L.</dc:creator>
<dc:creator>Sareen, D.</dc:creator>
<dc:creator>Svendsen, C. N.</dc:creator>
<dc:creator>Fraenkel, E.</dc:creator>
<dc:date>2023-09-13</dc:date>
<dc:identifier>doi:10.1101/2023.09.11.557005</dc:identifier>
<dc:title><![CDATA[Disease related changes in ATAC-Seq of more than 450 iPSC-derived motor neuron lines from ALS patients and controls]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.12.556572v1?rss=1">
<title>
<![CDATA[
Plasma membrane abundance dictates phagocytic capacity and functional crosstalk in myeloid cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.12.556572v1?rss=1</link>
<description><![CDATA[
Professional phagocytes like neutrophils and macrophages tightly control what they eat, how much they eat, and when they move after eating. We show that plasma membrane abundance is a key arbiter of these cellular behaviors. Neutrophils and macrophages lacking the G-protein subunit G{beta}4 exhibit profound plasma membrane expansion due to enhanced production of sphingolipids. This increased membrane allocation dramatically enhances phagocytosis of bacteria, fungus, apoptotic corpses, and cancer cells. G{beta}4 deficient neutrophils are also defective in the normal inhibition of migration following cargo uptake. In G{beta}4 knockout mice, myeloid cells exhibit enhanced phagocytosis of inhaled fungal conidia in the lung but also increased trafficking of engulfed pathogens to other organs. These results reveal an unexpected, biophysical control mechanism lying at the heart of myeloid functional decision-making.
]]></description>
<dc:creator>Winer, B. Y.</dc:creator>
<dc:creator>Settle, A. H.</dc:creator>
<dc:creator>Yakimov, A. M.</dc:creator>
<dc:creator>Jeronimo, C.</dc:creator>
<dc:creator>Lazarovt, T.</dc:creator>
<dc:creator>Tipping, M.</dc:creator>
<dc:creator>Saoi, M.</dc:creator>
<dc:creator>Sawh, A.</dc:creator>
<dc:creator>Sepp, A.-L. L.</dc:creator>
<dc:creator>Galiano, M.</dc:creator>
<dc:creator>Perry, J. S. A.</dc:creator>
<dc:creator>Wong, Y. Y.</dc:creator>
<dc:creator>Geissmann, F.</dc:creator>
<dc:creator>Cross, J.</dc:creator>
<dc:creator>Zhou, T.</dc:creator>
<dc:creator>Kam, L. C.</dc:creator>
<dc:creator>Pasoli, H. A.</dc:creator>
<dc:creator>Hohl, T.</dc:creator>
<dc:creator>Cyster, J. G.</dc:creator>
<dc:creator>Weiner, O.</dc:creator>
<dc:creator>Huse, M.</dc:creator>
<dc:date>2023-09-13</dc:date>
<dc:identifier>doi:10.1101/2023.09.12.556572</dc:identifier>
<dc:title><![CDATA[Plasma membrane abundance dictates phagocytic capacity and functional crosstalk in myeloid cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.12.556998v1?rss=1">
<title>
<![CDATA[
Protein phosphatase 1 regulates core PCP signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.12.556998v1?rss=1</link>
<description><![CDATA[
AbstractPCP signaling polarizes epithelial cells within the plane of an epithelium. Core PCP signaling components adopt asymmetric subcellular localizations within cells to both polarize and coordinate polarity between cells. Achieving subcellular asymmetry requires additional effectors, including some mediating post-translational modifications of core components. Identification of such proteins is challenging due to pleiotropy. We used mass spectrometry-based proximity labeling proteomics to identify such regulators in the Drosophila wing. We identified the catalytic subunit of Protein Phosphatase1, Pp1-87B, and show that it regulates core protein polarization. Pp1-87B interacts with the core protein Van Gogh and at least one Serine/Threonine kinase, Dco/CKIc, that is known to regulate PCP. Pp1-87B modulates Van Gogh subcellular localization and directs its dephosphorylation in vivo. PNUTS, a Pp1 regulatory subunit, also modulates PCP. While the direct substrate(s) of Pp1-87B in control of PCP is not known, our data support the model that cycling between phosphorylated and unphosphorylated forms of one or more core PCP components may regulate acquisition of asymmetry. Finally, our screen serves as a resource for identifying additional regulators of PCP signaling.
]]></description>
<dc:creator>Song, S.</dc:creator>
<dc:creator>Cho, B.</dc:creator>
<dc:creator>Weiner, A. T.</dc:creator>
<dc:creator>Nissen, S. B.</dc:creator>
<dc:creator>Naharros, I. O.</dc:creator>
<dc:creator>Bosch, P. S.</dc:creator>
<dc:creator>Suyama, K.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>He, L.</dc:creator>
<dc:creator>Svinkina, T.</dc:creator>
<dc:creator>Udeshi, N.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Perrimon, N.</dc:creator>
<dc:creator>Axelrod, J. D.</dc:creator>
<dc:date>2023-09-13</dc:date>
<dc:identifier>doi:10.1101/2023.09.12.556998</dc:identifier>
<dc:title><![CDATA[Protein phosphatase 1 regulates core PCP signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.11.557127v1?rss=1">
<title>
<![CDATA[
Discovery of a functionally selective serotonin 5-HT1A receptor agonist for the treatment of pain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.11.557127v1?rss=1</link>
<description><![CDATA[
The G protein-coupled serotonin receptor 5-HT1AR mediates antinociception and may serve as a valuable target for the treatment of pain. Starting from a chemical library, ST171, a bitopic chemotype activating 5-HT1AR was evolved. In vitro pharmacological investigations of ST171 revealed potent and selective Gi activation (EC50 = 0.3 nM), with marginal Gs and {beta}-arrestin recruitment. Preclinical studies in mice showed that ST171 was effective in acute and chronic (inflammatory and neuropathic) pain models, without causing sedation. Comparison of cryo-EM structures of a 5-HT1AR-Gi complex bound to the functionally biased agonist ST171, with a structure bound to the functionally balanced agonist befiradol, showed that both ligands bind to the same orthosteric site, but address different exo-sites. The individual poses are associated with ligand-specific helical dispositions and rearrangements of microdomains. Complementation of these studies with molecular dynamics simulations allowed us to derive structural features associated with ST171s functional selectivity, a phenomenon that may be crucial to the discovery of more effective and safe GPCR drugs.
]]></description>
<dc:creator>Ullrich, A.</dc:creator>
<dc:creator>Schneider, J.</dc:creator>
<dc:creator>Braz, J. M.</dc:creator>
<dc:creator>Neu, E.</dc:creator>
<dc:creator>Staffen, N.</dc:creator>
<dc:creator>Stanek, M.</dc:creator>
<dc:creator>Blahova, J.</dc:creator>
<dc:creator>Hove, T.</dc:creator>
<dc:creator>Albert, T.</dc:creator>
<dc:creator>Allikalt, A.</dc:creator>
<dc:creator>Lober, S.</dc:creator>
<dc:creator>Bhardwaj, K.</dc:creator>
<dc:creator>Rodriguez-Rosado, S.</dc:creator>
<dc:creator>Fink, E. A.</dc:creator>
<dc:creator>Rasmussen, T.</dc:creator>
<dc:creator>Hubner, H.</dc:creator>
<dc:creator>Inoue, A. A.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Basbaum, A.</dc:creator>
<dc:creator>Bottcher, B.</dc:creator>
<dc:creator>Weikert, D.</dc:creator>
<dc:creator>Gmeiner, P.</dc:creator>
<dc:date>2023-09-14</dc:date>
<dc:identifier>doi:10.1101/2023.09.11.557127</dc:identifier>
<dc:title><![CDATA[Discovery of a functionally selective serotonin 5-HT1A receptor agonist for the treatment of pain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.14.557527v1?rss=1">
<title>
<![CDATA[
circHIPK3 nucleates IGF2BP2 and functions as a competing endogenous RNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.14.557527v1?rss=1</link>
<description><![CDATA[
Circular RNAs (circRNAs) represent a class of widespread endogenous RNAs that regulate gene expression and thereby influence cell biological decisions with implications for the pathogenesis of several diseases. Here, we disclose a novel gene-regulatory role of circHIPK3 by combining analyses of large genomics datasets and mechanistic cell biological follow-up experiments. Specifically, we use temporal depletion of circHIPK3 or specific RNA binding proteins (RBPs) and identify several perturbed genes by RNA sequencing analyses. Using expression-coupled motif analyses of mRNA expression data from various knockdown experiments, we identify an 11-mer motif within circHIPK3, which is also enriched in genes that become downregulated upon circHIPK3 depletion. By mining eCLIP datasets, we find that the 11-mer motif constitutes a strong binding site for IGF2BP2 and validate this circHIPK3-IGF2BP2 interaction experimentally using RNA-immunoprecipitation and competition assays in bladder cancer cell lines. Our results suggest that circHIPK3 and IGF2BP2 mRNA targets compete for binding. Since the identified 11-mer motif found in circHIPK3 is enriched in upregulated genes following IGF2BP2 knockdown, and since IGF2BP2 depletion conversely globally antagonizes the effect of circHIPK3 knockdown on target genes, our results suggest that circHIPK3 can sequester IGF2BP2 as a competing endogenous RNA (ceRNA), leading to target mRNA stabilization. As an example of a circHIPK3-regulated gene, we focus on the STAT3 mRNA as a specific substrate of IGF2BP2 and validate that manipulation of circHIPK3 regulates IGF2BP2-STAT3 mRNA binding and thereby STAT3 mRNA levels. However, absolute copy number quantifications demonstrate that IGF2BP2 outnumbers circHIPK3 by orders of magnitude, which is inconsistent with a simple 1:1 ceRNA hypothesis. Instead, we show that circHIPK3 can nucleate multiple copies of IGF2BP2, potentially via phase separation, to produce IGF2BP2 condensates. Finally, we show that circHIPK3 expression correlates with overall survival of patients with bladder cancer. Our results are consistent with a model where relatively few cellular circHIPK3 molecules function as inducers of IGF2BP2 condensation thereby regulating STAT3 and other key factors for cell proliferation and potentially cancer progression.
]]></description>
<dc:creator>Okholm, T. L. H.</dc:creator>
<dc:creator>Kamstrup, A. B.</dc:creator>
<dc:creator>Nielsen, M. M.</dc:creator>
<dc:creator>Hollensen, A. K.</dc:creator>
<dc:creator>Graversgaard, M. L.</dc:creator>
<dc:creator>Kristensen, L. S.</dc:creator>
<dc:creator>Vang, S.</dc:creator>
<dc:creator>Park, S. S.</dc:creator>
<dc:creator>Yeo, G. W.</dc:creator>
<dc:creator>Dyrskjot, L.</dc:creator>
<dc:creator>Kjems, J.</dc:creator>
<dc:creator>Pedersen, J. S.</dc:creator>
<dc:creator>Damgaard, C. K.</dc:creator>
<dc:date>2023-09-14</dc:date>
<dc:identifier>doi:10.1101/2023.09.14.557527</dc:identifier>
<dc:title><![CDATA[circHIPK3 nucleates IGF2BP2 and functions as a competing endogenous RNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.14.557805v1?rss=1">
<title>
<![CDATA[
Determinants of DDX3X sensitivity uncovered using a helicase activity in translation reporter 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.14.557805v1?rss=1</link>
<description><![CDATA[
DDX3X regulates the translation of a subset of human transcripts containing complex 5' untranslated regions (5' UTRs). In this study we developed the helicase activity reporter for translation (HART) which uses DDX3X-sensitive 5' UTRs to measure DDX3X mediated translational activity in cells. To dissect the structural underpinnings of DDX3X dependent translation, we first used SHAPE-MaP to determine the secondary structures present in DDX3X-sensitive 5' UTRs and then employed HART to investigate how their perturbation impacts DDX3X-sensitivity. Additionally, we identified residues 38-44 as potential mediators of DDX3Xs interaction with the translational machinery. HART revealed that both DDX3Xs association with the ribosome complex as well as its helicase activity are required for its function in promoting the translation of DDX3X-sensitive 5' UTRs. These findings suggest DDX3X plays a crucial role regulating translation through its interaction with the translational machinery during ribosome scanning, and establish the HART reporter as a robust, lentivirally encoded measurement of DDX3X-dependent translation in cells.
]]></description>
<dc:creator>Wilkins, K.</dc:creator>
<dc:creator>Schroeder, T.</dc:creator>
<dc:creator>Gu, S.</dc:creator>
<dc:creator>Revalde, J.</dc:creator>
<dc:creator>Floor, S. N.</dc:creator>
<dc:date>2023-09-14</dc:date>
<dc:identifier>doi:10.1101/2023.09.14.557805</dc:identifier>
<dc:title><![CDATA[Determinants of DDX3X sensitivity uncovered using a helicase activity in translation reporter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.15.556569v1?rss=1">
<title>
<![CDATA[
CCR2-targeting pepducins reduce T cell-nociceptor interaction driving bone cancer pain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.15.556569v1?rss=1</link>
<description><![CDATA[
Inhibition of the CCL2/CCR2 chemokine signaling represents a promising avenue for the development of non-opioid pain treatment, particularly for painful bone metastases. To investigate the involvement of CCR2 in cancer-induced bone pain, we generated and characterized the functional activities of a novel cell-penetrating pepducin, namely PP101, acting as an intracellular negative allosteric modulator of CCR2. In vivo, PP101 was effective in relieving neuropathic and bone cancer pain. By targeting CCR2, PP101 reduced bone cancer pain by preventing infiltration of CD4+ and CD8+ T cells and by decreasing the neuroimmune communication network within the dorsal root ganglia. Importantly, reduced neuroinflammatory milieu in the dorsal root ganglia induced by PP101 did not result in deleterious tumor progression or behavioral adverse effects. Thus, targeting the neuroimmune crosstalk through allosteric inhibition of CCR2 may represent an effective and safe avenue for the management of bone cancer pain.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=147 SRC="FIGDIR/small/556569v1_ufig1.gif" ALT="Figure 1">
View larger version (41K):
org.highwire.dtl.DTLVardef@15c2d43org.highwire.dtl.DTLVardef@9fa40corg.highwire.dtl.DTLVardef@cb457dorg.highwire.dtl.DTLVardef@13f36e1_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LIBreast cancer bone metastases induce pain by activating CCR2 on sensory neurons.
C_LIO_LIDRG-infiltrating CD4+ and CD8+ T cells promote the development of bone cancer pain.
C_LIO_LICCR2 inhibition by PP101 suppresses DRG neuroinflammation and neuronal excitability.
C_LIO_LIPP101 alleviates bone cancer pain without behavioral or physiological side effects.
C_LI
]]></description>
<dc:creator>Midavaine, E.</dc:creator>
<dc:creator>Brouillette, R. R.</dc:creator>
<dc:creator>Theberge, E.</dc:creator>
<dc:creator>Mona, C. E.</dc:creator>
<dc:creator>Kashem, S. W.</dc:creator>
<dc:creator>Cote, J.</dc:creator>
<dc:creator>Zeugin, V.</dc:creator>
<dc:creator>Besserer-Offroy, E.</dc:creator>
<dc:creator>Longpre, J.-M.</dc:creator>
<dc:creator>Marsault, E.</dc:creator>
<dc:creator>Sarret, P.</dc:creator>
<dc:date>2023-09-15</dc:date>
<dc:identifier>doi:10.1101/2023.09.15.556569</dc:identifier>
<dc:title><![CDATA[CCR2-targeting pepducins reduce T cell-nociceptor interaction driving bone cancer pain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.15.558002v1?rss=1">
<title>
<![CDATA[
Ventromedial Prefrontal Cortex and Basolateral Amygdala Projections to Ventromedial Striatum Encode Active Avoidance Behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.15.558002v1?rss=1</link>
<description><![CDATA[
Active avoidance is a fundamental defensive behavior, defined as performing a voluntary action to escape a noxious stimulus. Active avoidance serves as an adaptive response to threat, ensuring safety. In individuals with anxiety disorders and post-traumatic stress disorder, avoidance behavior can become maladaptive when even safe situations are avoided, at great psychosocial cost. Corticostriatal and amygdalostriatal pathways have been implicated in active avoidance, though their role in learning and expression of this behavior is not fully understood. Projections from the ventromedial prefrontal cortex (vmPFC) to the ventromedial striatum (VMS) have been shown to regulate a variety of defensive behaviors; however, their role in active avoidance has not previously been examined. Additionally, basolateral amygdala (BLA) projections to the VMS have been shown to promote active avoidance in studies utilizing pharmacological inactivation and optogenetic inhibition, though in vivo real time neural activity in this pathway has not been recorded during active avoidance learning. Here we utilized fiber photometry for in vivo recordings of neural activity in vmPFC-VMS and BLA-VMS projections during active avoidance learning and expression. We implemented a two-way signaled active avoidance paradigm in which a light cue served as the conditioned stimulus (CS) signaling an impending foot shock. We examined changes in neural activity in these two projections during CS presentation, onset of active avoidance, and conditioned freezing. We found that vmPFC-VMS projections develop learning-related increases in activity at CS onset across training, while BLA-VMS projections do not show learning-related encoding of the CS. Additionally, we found that both vmPFC-VMS and BLA-VMS projections develop an increase in activity at avoidance onset. No changes in neural activity were observed during cued freezing in either vmPFC-VMS or BLA-VMS projections. Together these results indicate that vmPFC-VMS projections encode both CS and avoidance, while BLA-VMS projections may simply encode avoidance. Finally, we utilized optogenetic inhibition of vmPFC-VMS projections during the CS to investigate the necessity of this pathway for expression of active avoidance behavior. We found that inhibition of vmPFC-VMS projections attenuates learned active avoidance behavior, indicating that activity in this pathway is required for proper active avoidance. In summary, our results demonstrate task-relevant encoding of active avoidance behavior in vmPFC-VMS and BLA-VMS projections.
]]></description>
<dc:creator>Bullock, T. E.</dc:creator>
<dc:creator>Gunaydin, L. A.</dc:creator>
<dc:date>2023-09-15</dc:date>
<dc:identifier>doi:10.1101/2023.09.15.558002</dc:identifier>
<dc:title><![CDATA[Ventromedial Prefrontal Cortex and Basolateral Amygdala Projections to Ventromedial Striatum Encode Active Avoidance Behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.16.558090v1?rss=1">
<title>
<![CDATA[
Multiplexed CRISPR gene editing in primary human islet cells with Cas9 ribonucleoprotein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.16.558090v1?rss=1</link>
<description><![CDATA[
Successful genome editing in primary human islets could reveal features of the genetic regulatory landscape underlying {beta} cell function and diabetes risk. Here, we describe a CRISPR-based strategy to interrogate functions of predicted regulatory DNA elements using electroporation of a complex of Cas9 ribonucleoprotein (Cas9 RNP) and guide RNAs into primary human islet cells. We successfully targeted coding regions including the PDX1 exon 1, and non-coding DNA linked to diabetes susceptibility. CRISPR/Cas9 RNP approaches revealed genetic targets of regulation by DNA elements containing candidate diabetes risk SNPs, including an in vivo enhancer of the MPHOSPH9 gene. CRISPR/Cas9 RNP multiplexed targeting of two cis-regulatory elements linked to diabetes risk in PCSK1, which encodes an endoprotease crucial for insulin processing, also demonstrated efficient simultaneous editing of PCSK1 regulatory elements, resulting in impaired {beta} cell PCSK1 regulation and insulin secretion. Multiplex CRISPR/Cas9 RNP provides powerful approaches to investigate and elucidate human islet cell gene regulation in health and diabetes.
]]></description>
<dc:creator>Bevacqua, R. J.</dc:creator>
<dc:creator>Zhao, W.</dc:creator>
<dc:creator>Merheb, E.</dc:creator>
<dc:creator>Kim, S. H.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Gloyn, A. L.</dc:creator>
<dc:creator>Kim, S. K.</dc:creator>
<dc:date>2023-09-17</dc:date>
<dc:identifier>doi:10.1101/2023.09.16.558090</dc:identifier>
<dc:title><![CDATA[Multiplexed CRISPR gene editing in primary human islet cells with Cas9 ribonucleoprotein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.17.557749v1?rss=1">
<title>
<![CDATA[
Gene regulatory network inference from CRISPR perturbations in primary CD4+ T cells elucidates the genomic basis of immune disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.17.557749v1?rss=1</link>
<description><![CDATA[
The effects of genetic variation on complex traits act mainly through changes in gene regulation. Although many genetic variants have been linked to target genes in cis, the trans-regulatory cascade mediating their effects remains largely uncharacterized. Mapping trans-regulators based on natural genetic variation, including eQTL mapping, has been challenging due to small effects. Experimental perturbation approaches offer a complementary and powerful approach to mapping trans-regulators. We used CRISPR knockouts of 84 genes in primary CD4+ T cells to perturb an immune cell gene network, targeting both inborn error of immunity (IEI) disease transcription factors (TFs) and background TFs matched in constraint and expression level, but without a known immune disease association. We developed a novel Bayesian structure learning method called Linear Latent Causal Bayes (LLCB) to estimate the gene regulatory network from perturbation data and observed 211 directed edges among the genes which could not be detected in existing CD4+ trans-eQTL data. We used LLCB to characterize the differences between the IEI and background TFs, finding that the gene groups were highly interconnected, but that IEI TFs were much more likely to regulate immune cell specific pathways and immune GWAS genes. We further characterized nine coherent gene programs based on downstream effects of the TFs and linked these modules to regulation of GWAS genes, finding that canonical JAK-STAT family members are regulated by KMT2A, a global epigenetic regulator. These analyses reveal the trans-regulatory cascade from upstream epigenetic regulator to intermediate TFs to downstream effector cytokines and elucidate the logic linking immune GWAS genes to key signaling pathways.
]]></description>
<dc:creator>Weinstock, J. S.</dc:creator>
<dc:creator>Arce, M. M.</dc:creator>
<dc:creator>Freimer, J. W.</dc:creator>
<dc:creator>Ota, M.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Battle, A.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:date>2023-09-17</dc:date>
<dc:identifier>doi:10.1101/2023.09.17.557749</dc:identifier>
<dc:title><![CDATA[Gene regulatory network inference from CRISPR perturbations in primary CD4+ T cells elucidates the genomic basis of immune disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.18.558350v1?rss=1">
<title>
<![CDATA[
Higher-order combinatorial chromatin perturbations by engineered CRISPR-Cas12a for functional genomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.18.558350v1?rss=1</link>
<description><![CDATA[
Multiplexed genetic perturbations are critical for testing functional interactions among coding or non-coding genetic elements. Compared to double-stranded DNA cutting, repressive chromatin formation using CRISPR interference (CRISPRi) avoids genotoxicity and is more effective for perturbing non-coding regulatory elements in pooled assays. However, current CRISPRi pooled screening approaches are limited to targeting 1-3 genomic sites per cell. To develop a tool for higher-order (>3) combinatorial targeting of genomic sites with CRISPRi in functional genomics screens, we engineered an Acidaminococcus Cas12a variant -- referred to as multiplexed transcriptional interference AsCas12a (multiAsCas12a). multiAsCas12a incorporates a key mutation, R1226A, motivated by the hypothesis of nicking-induced stabilization of the ribonucleoprotein:DNA complex for improving CRISPRi activity. multiAsCas12a significantly outperforms prior state-of-the-art Cas12a variants in combinatorial CRISPRi targeting using high-order multiplexed arrays of lentivirally transduced CRISPR RNAs (crRNA), including in high-throughput pooled screens using 6-plex crRNA array libraries. Using multiAsCas12a CRISPRi, we discover new enhancer elements and dissect the combinatorial function of cis-regulatory elements. These results instantiate a group testing framework for efficiently surveying potentially numerous combinations of chromatin perturbations for biological discovery and engineering.
]]></description>
<dc:creator>Hsiung, C. C.- S.</dc:creator>
<dc:creator>Wilson, C. M.</dc:creator>
<dc:creator>Sambold, N. A.</dc:creator>
<dc:creator>Dai, R.</dc:creator>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>Misiukiewicz, S.</dc:creator>
<dc:creator>Arab, A.</dc:creator>
<dc:creator>Teyssier, N.</dc:creator>
<dc:creator>O'Loughlin, T.</dc:creator>
<dc:creator>Cofsky, J.</dc:creator>
<dc:creator>Shi, J.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:date>2023-09-18</dc:date>
<dc:identifier>doi:10.1101/2023.09.18.558350</dc:identifier>
<dc:title><![CDATA[Higher-order combinatorial chromatin perturbations by engineered CRISPR-Cas12a for functional genomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.18.558356v1?rss=1">
<title>
<![CDATA[
De novo designed Hsp70 activator dissolves intracellular condensates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.18.558356v1?rss=1</link>
<description><![CDATA[
Protein quality control (PQC) is carried out in part by the chaperone Hsp70, in concert with adapters of the J-domain protein (JDP) family. The JDPs, also called Hsp40s, are thought to recruit Hsp70 into complexes with specific client proteins. However, the molecular principles regulating this process are not well understood. We describe the de novo design of a set of Hsp70 binding proteins that either inhibited or stimulated Hsp70s ATPase activity; a stimulating design promoted the refolding of denatured luciferase in vitro, similar to native JDPs. Targeting of this design to intracellular condensates resulted in their nearly complete dissolution. The designs inform our understanding of chaperone structure-function relationships and provide a general and modular way to target PQC systems to condensates and other cellular targets.
]]></description>
<dc:creator>Zhang, J. Z.</dc:creator>
<dc:creator>Greenwood, N.</dc:creator>
<dc:creator>Hernandez, J.</dc:creator>
<dc:creator>Cuperus, J. T.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Ryder, B. D.</dc:creator>
<dc:creator>Queitsch, C.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2023-09-19</dc:date>
<dc:identifier>doi:10.1101/2023.09.18.558356</dc:identifier>
<dc:title><![CDATA[De novo designed Hsp70 activator dissolves intracellular condensates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.19.558439v1?rss=1">
<title>
<![CDATA[
Sex Chromosomes and Gonads Shape the Sex-Biased Transcriptomic Landscape in Tlr7-Mediated Demyelination During Aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.19.558439v1?rss=1</link>
<description><![CDATA[
Demyelination occurs in aging and associated diseases, including Alzheimers disease. Several of these diseases exhibit sex differences in prevalence and severity. Biological sex primarily stems from sex chromosomes and gonads releasing sex hormones. To dissect mechanisms underlying sex differences in demyelination of aging brains, we constructed a transcriptomic atlas of cell type-specific responses to illustrate how sex chromosomes, gonads, and their interaction shape responses to demyelination. We found that sex-biased oligodendrocyte and microglial responses are driven by interaction of sex chromosomes and gonads prior to myelin loss. Post demyelination, sex chromosomes mainly guide microglial responses, while gonadal composition influences oligodendrocyte signaling. Significantly, ablation of the X-linked gene Toll-like receptor 7 (Tlr7), which exhibited sex-biased expression during demyelination, abolished the sex-biased responses and protected against demyelination.

One-sentence summaryCell type-specific processes underlying aged demyelination are sex-biased and mediated by Tlr7.
]]></description>
<dc:creator>Lopez-Lee, C.</dc:creator>
<dc:creator>Kodama, L.</dc:creator>
<dc:creator>Fan, L.</dc:creator>
<dc:creator>Wong, M. Y.</dc:creator>
<dc:creator>Foxe, N. R.</dc:creator>
<dc:creator>Ijaz, L.</dc:creator>
<dc:creator>Yu, F.</dc:creator>
<dc:creator>Ye, P.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Norman, K.</dc:creator>
<dc:creator>Torres, E. R.</dc:creator>
<dc:creator>Kim, R.</dc:creator>
<dc:creator>Mousa, G. A.</dc:creator>
<dc:creator>Dubal, D.</dc:creator>
<dc:creator>Liddelow, S.</dc:creator>
<dc:creator>Luo, W.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:date>2023-09-21</dc:date>
<dc:identifier>doi:10.1101/2023.09.19.558439</dc:identifier>
<dc:title><![CDATA[Sex Chromosomes and Gonads Shape the Sex-Biased Transcriptomic Landscape in Tlr7-Mediated Demyelination During Aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.20.558582v1?rss=1">
<title>
<![CDATA[
Circulating Platelets Modulate Oligodendrocyte Progenitor Cell Differentiation During Remyelination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.20.558582v1?rss=1</link>
<description><![CDATA[
Revealing unknown cues that regulate oligodendrocyte progenitor cell (OPC) function in remyelination is important to optimise the development of regenerative therapies for multiple sclerosis (MS). Platelets are present in chronic non-remyelinated lesions of MS and an increase in circulating platelets has been described in experimental autoimmune encephalomyelitis (EAE) mice, an animal model for MS. However, the contribution of platelets to remyelination remains unexplored. Here we show platelet aggregation in proximity to OPCs in areas of experimental demyelination. Partial depletion of circulating platelets impaired OPC differentiation and remyelination, without altering blood-brain barrier stability and neuroinflammation. Transient exposure to platelets enhanced OPC differentiation in vitro, whereas sustained exposure suppressed this effect. In a mouse model of thrombocytosis (CALRHET), there was a sustained increase in platelet aggregation together with a reduction of newly-generated oligodendrocytes following toxin-induced demyelination. These findings reveal a complex bimodal contribution of platelet to remyelination and provide insights into remyelination failure in MS.
]]></description>
<dc:creator>Philp, A. R.</dc:creator>
<dc:creator>Reyes, C. R.</dc:creator>
<dc:creator>Mansilla, J.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Valenzuela-Krugmann, C.</dc:creator>
<dc:creator>Rawji, K. S.</dc:creator>
<dc:creator>Martinez, G. A. G.</dc:creator>
<dc:creator>Dimas, P.</dc:creator>
<dc:creator>Hinrichsen, B.</dc:creator>
<dc:creator>Ulloa-Leal, C.</dc:creator>
<dc:creator>Waller, A. K.</dc:creator>
<dc:creator>Bessa de Sousa, D. M.</dc:creator>
<dc:creator>Castro, M. A.</dc:creator>
<dc:creator>Aigner, L.</dc:creator>
<dc:creator>Ehrenfeld, P.</dc:creator>
<dc:creator>Silva, M. E.</dc:creator>
<dc:creator>Kazanis, I.</dc:creator>
<dc:creator>Ghevaert, C.</dc:creator>
<dc:creator>Franklin, R. J. M.</dc:creator>
<dc:creator>Rivera, F. J.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.20.558582</dc:identifier>
<dc:title><![CDATA[Circulating Platelets Modulate Oligodendrocyte Progenitor Cell Differentiation During Remyelination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.20.558726v1?rss=1">
<title>
<![CDATA[
Epithelial zonation along the mouse and human small intestine defines five discrete metabolic domains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.20.558726v1?rss=1</link>
<description><![CDATA[
A key aspect of nutrient absorption is the exquisite division of labor across the length of the small intestine, with individual classes of micronutrients taken up at different positions. For millennia, the small intestine was thought to comprise three segments with indefinite borders: the duodenum, jejunum, and ileum. By examining fine-scale longitudinal segmentation of the mouse and human small intestines, we identified transcriptional signatures and upstream regulatory factors that define five domains of nutrient absorption, distinct from the three traditional sections. Spatially restricted expression programs were most prominent in nutrient-absorbing enterocytes but initially arose in intestinal stem cells residing in three regional populations. While a core signature was maintained across mice and humans with different diets and environments, domain properties were influenced by dietary changes. We established the functions of Ppar- and Cdx1 in patterning lipid metabolism in distal domains and generated a predictive model of additional transcription factors that direct domain identity. Molecular domain identity can be detected with machine learning, representing the first systematic method to computationally identify specific intestinal regions in mice. These findings provide a foundational framework for the identity and control of longitudinal zonation of absorption along the proximal:distal small intestinal axis.
]]></description>
<dc:creator>Zwick, R. K.</dc:creator>
<dc:creator>Kasparek, P.</dc:creator>
<dc:creator>Palikuqi, B.</dc:creator>
<dc:creator>Viragova, S.</dc:creator>
<dc:creator>Weichselbaum, L.</dc:creator>
<dc:creator>McGinnis, C. S.</dc:creator>
<dc:creator>McKinley, K. L.</dc:creator>
<dc:creator>Rathnayake, A.</dc:creator>
<dc:creator>Vaka, D.</dc:creator>
<dc:creator>Nguyen, V.</dc:creator>
<dc:creator>Trentesaux, C.</dc:creator>
<dc:creator>Reyes, E.</dc:creator>
<dc:creator>Gupta, A.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:creator>Locksley, R. M.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Itzkovitz, S.</dc:creator>
<dc:creator>Boffelli, D.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.20.558726</dc:identifier>
<dc:title><![CDATA[Epithelial zonation along the mouse and human small intestine defines five discrete metabolic domains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.21.558893v1?rss=1">
<title>
<![CDATA[
PAI-1 Deficiency Drives Pulmonary Vascular Smooth Muscle Remodeling and Pulmonary Hy-pertension 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.21.558893v1?rss=1</link>
<description><![CDATA[
Pulmonary arterial hypertension (PAH) is a progressive and potentially a rapidly fatal disease characterized by vasoconstriction and remodeling of small pulmonary arteries (PA) leading to increased pulmonary vascular resistance and right heart failure. Central to the remodeling process is a switch of the smooth muscle cells in small PAs (PASMC) to a proliferative, apoptosis-resistant phenotype.

There is reason to suspect that the plasminogen activator system may play an important role in the remodeling program in PAH based on its roles in vascular post-injury restenosis, fibrosis, angiogenesis and tumorigenesis. Plasminogen activator inhibitor-1 (PAI-1) is the primary physiological inhibitor of the plasminogen activators - urokinase-type and tissue-type (uPA and tPA, respectively). Immunohisto- chemical and immunoblot analyses revealed that PAI-1 was deficient in smooth muscle areas of small remodeled PAs and early-passage PASMC from subjects with PAH compared to non-PAH controls. PAI1-/- male and female mice developed spontaneous pulmonary vascular remodeling and pulmonary hypertension (PH) as evidenced by significant increase in PA medial thickness, systolic right ventricular pressure, and right ventricular hypertrophy. Lastly, the uPA inhibitors upamostat (WX-671) and amiloride analog BB2-30F down-regulated mTORC1 and SMAD3, restored PAI-1 levels, reduced proliferation, and induced apoptosis in human PAH PASMC. We examined the effect of inhibition of uPA catalytic activity by BB2-30F on the development of SU5416/Hypoxia (SuHx)-induced PH in mice. Vehicletreated SuHx-exposed mice had up-regulated mTORC1 in small PAs, developed pulmonary vascular remodeling and PH, as evidenced by significant increase of PA MT, sRVP, RV hypertrophy, and a significant decrease in the pulmonary artery acceleration time/pulmonary ejection time (PAAT/PET) ratio compared to age- and sex-matched normoxia controls, whereas BB2-30F-treated group was protected from all these pathological changes. Taken together, our data strongly suggest that PAI-1 down- regulation in PASMC from human PAH lungs promotes PASMC hyper-proliferation, remodeling, and spontaneous PH due to unopposed uPA activation. Further studies are needed to determine the potential benefits of targeting the PAI-1/uPA imbalance to attenuate the progression and/or reverse pulmonary vascular remodeling and PH.
]]></description>
<dc:creator>Kudryashova, T. V.</dc:creator>
<dc:creator>Zaitsev, S. V.</dc:creator>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>Buckley, B. J.</dc:creator>
<dc:creator>McGuckin, J. P.</dc:creator>
<dc:creator>Goncharov, D.</dc:creator>
<dc:creator>Zhyvylo, I.</dc:creator>
<dc:creator>Lin, D.</dc:creator>
<dc:creator>Newcomb, G.</dc:creator>
<dc:creator>Piper, B.</dc:creator>
<dc:creator>Bogamuwa, S.</dc:creator>
<dc:creator>Saiyed, A.</dc:creator>
<dc:creator>Teos, L.</dc:creator>
<dc:creator>Ranson, M.</dc:creator>
<dc:creator>Wolters, P.</dc:creator>
<dc:creator>Kelso, M. J.</dc:creator>
<dc:creator>Poncz, M.</dc:creator>
<dc:creator>DeLisser, H. M.</dc:creator>
<dc:creator>Cines, D. B.</dc:creator>
<dc:creator>Goncharova, E. A.</dc:creator>
<dc:creator>Farkas, L.</dc:creator>
<dc:creator>Stepanova, V.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.21.558893</dc:identifier>
<dc:title><![CDATA[PAI-1 Deficiency Drives Pulmonary Vascular Smooth Muscle Remodeling and Pulmonary Hy-pertension]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.22.558671v1?rss=1">
<title>
<![CDATA[
Cryo-EM structure of Alzheimer disease tau filaments with PET ligand MK-6240 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.22.558671v1?rss=1</link>
<description><![CDATA[
Positron Emission Tomography (PET) ligands have advanced Alzheimers disease (AD) diagnosis and treatment. Using autoradiography and cryo-EM, we identified AD brain tissue with elevated tau burden, purified filaments, and determined the structure of second-generation high avidity PET ligand MK-6240 at 2.31 [A] resolution, which bound at a 1:1 ratio within the cleft of tau paired-helical filament (PHF), engaging with glutamine 351, lysine K353, and isoleucine 360. This information elucidates the basis of MK-6240 PET in quantifying PHF deposits in AD and may facilitate the structure-based design of superior ligands against tau amyloids.
]]></description>
<dc:creator>Kunach, P.</dc:creator>
<dc:creator>Vaquer-Alicea, J.</dc:creator>
<dc:creator>Smith, M. S.</dc:creator>
<dc:creator>Hopewell, R.</dc:creator>
<dc:creator>Monistrol, J.</dc:creator>
<dc:creator>Moquin, L.</dc:creator>
<dc:creator>Therriault, J.</dc:creator>
<dc:creator>Tissot, C.</dc:creator>
<dc:creator>Rahmouni, N.</dc:creator>
<dc:creator>Massarweh, G.</dc:creator>
<dc:creator>Soucy, J.-P.</dc:creator>
<dc:creator>Guiot, M.-C.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Rosa-Neto, P. K.</dc:creator>
<dc:creator>Diamond, M. I.</dc:creator>
<dc:creator>Shahmoradian, S. H.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.22.558671</dc:identifier>
<dc:title><![CDATA[Cryo-EM structure of Alzheimer disease tau filaments with PET ligand MK-6240]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.22.559056v1?rss=1">
<title>
<![CDATA[
Single cell analysis of dup15q syndrome reveals developmental and postnatal molecular changes in autism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.22.559056v1?rss=1</link>
<description><![CDATA[
Duplication 15q (dup15q) syndrome is the most common genetic cause of autism spectrum disorder (ASD). Due to a higher genetic and phenotypic homogeneity compared to idiopathic autism, dup15q syndrome provides a well-defined setting to investigate ASD mechanisms. Previous bulk gene expression studies identified shared molecular changes in ASD. However, how cell type specific changes compare across different autism subtypes and how they change during development is largely unknown. In this study, we used single cell and single nucleus mRNA sequencing of dup15q cortical organoids from patient iPSCs, as well as post-mortem patient brain samples. We find cell-type specific dysregulated programs that underlie dup15q pathogenesis, which we validate by spatial resolved transcriptomics using brain tissue samples. We find degraded identity and vulnerability of deep-layer neurons in fetal stage organoids and highlight increased molecular burden of postmortem upper-layer neurons implicated in synaptic signaling, a finding shared between idiopathic ASD and dup15q syndrome. Gene co-expression network analysis of organoid and postmortem excitatory neurons uncovers modules enriched with autism risk genes. Organoid developmental modules were involved in transcription regulation via chromatin remodeling, while postmortem modules were associated with synaptic transmission and plasticity. The findings reveal a shifting landscape of ASD cellular vulnerability during brain development.
]]></description>
<dc:creator>Perez, Y.</dc:creator>
<dc:creator>Velmeshev, D.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>White, M.</dc:creator>
<dc:creator>Siebert, C.</dc:creator>
<dc:creator>Baltazar, J.</dc:creator>
<dc:creator>Dutton, N. G.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Haeussler, M.</dc:creator>
<dc:creator>Chamberlain, S.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.22.559056</dc:identifier>
<dc:title><![CDATA[Single cell analysis of dup15q syndrome reveals developmental and postnatal molecular changes in autism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.20.558707v1?rss=1">
<title>
<![CDATA[
GIA: A genome interval arithmetic toolkit for high performance interval set operations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.20.558707v1?rss=1</link>
<description><![CDATA[
MotivationThis study addresses the pressing need for efficient interval techniques in processing vast genomic datasets, such as those generated by ChIP-seq, RNA-seq, and whole-genome sequencing. Intervals are critical in characterizing biological features, necessitating streamlined interval manipulation for meaningful insights. Existing tools often struggle with memory and runtime requirements when managing extensive genomic region arithmetic.

ResultsThe study introduces GIA (Genomic Interval Arithmetic) and BEDRS, a novel command-line tool and a rust library that significantly enhance the performance of genomic interval analysis. GIA outperforms existing tools like BEDOPS, BEDTools, and GenomicRanges by a factor of 2x to 20x across a range of operations. These advances enable researchers to perform genomic interval operations more efficiently, drastically reducing computational time and resource requirements in high-throughput genomic sequencing analysis.

Availability and Implementation- https://github.com/noamteyssier/gia
- https://github.com/noamteyssier/bedrs
]]></description>
<dc:creator>Teyssier, N.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2023-09-23</dc:date>
<dc:identifier>doi:10.1101/2023.09.20.558707</dc:identifier>
<dc:title><![CDATA[GIA: A genome interval arithmetic toolkit for high performance interval set operations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.21.558743v1?rss=1">
<title>
<![CDATA[
Germline-targeting chimpanzee SIV Envelopes induce V2-apex broadly neutralizing-like B cell precursors in a rhesus macaque infection model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.21.558743v1?rss=1</link>
<description><![CDATA[
Eliciting broadly neutralizing antibodies-(bnAbs) remains a major goal of HIV-1 vaccine research. Previously, we showed that a soluble chimpanzee SIV Envelope-(Env) trimer, MT145K, bound several human V2-apex bnAb-precursors and stimulated an appropriate response in V2-apex bnAb precursor-expressing knock-in mice. Here, we tested the immunogenicity of three MT145 variants (MT145, MT145K, MT145K.dV5) expressed as chimeric simian-chimpanzee-immunodeficiency-viruses-(SCIVs) in rhesus macaques-(RMs). All three viruses established productive infections with high setpoint vRNA titers. RMs infected with the germline-targeting SCIV_MT145K and SCIV_MT145K.dV5 exhibited larger and more clonally expanded B cell lineages featuring long anionic heavy chain complementary-determining-regions-(HCDR3s) compared with wildtype SCIV_MT145. Moreover, antigen-specific B cell analysis revealed enrichment for long-CDHR3-bearing antibodies in SCIV_MT145K.dV5 infected animals with paratope features resembling prototypic V2-apex bnAbs and their precursors. Although none of the animals developed bnAbs, these results show that germline-targeting SCIVs can activate and preferentially expand B cells expressing V2-apex bnAb-like precursors, the first step in bnAb elicitation.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=180 HEIGHT=200 SRC="FIGDIR/small/558743v1_ufig1.gif" ALT="Figure 1">
View larger version (57K):
org.highwire.dtl.DTLVardef@17673c8org.highwire.dtl.DTLVardef@1ab61a8org.highwire.dtl.DTLVardef@98781forg.highwire.dtl.DTLVardef@1f63e9b_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Musharrafieh, R.</dc:creator>
<dc:creator>Safonova, Y.</dc:creator>
<dc:creator>Song, G.</dc:creator>
<dc:creator>Roark, R. S.</dc:creator>
<dc:creator>Lee, F.-H.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Hurtado, J.</dc:creator>
<dc:creator>Yong, P.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Russell, R. M.</dc:creator>
<dc:creator>Ding, W.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Rando, J.</dc:creator>
<dc:creator>Murphy, A. I.</dc:creator>
<dc:creator>Lindemuth, E.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Connell, A. J.</dc:creator>
<dc:creator>Lee, W.-H.</dc:creator>
<dc:creator>Mishra, N.</dc:creator>
<dc:creator>Avillion, G.</dc:creator>
<dc:creator>He, W.</dc:creator>
<dc:creator>Callaghan, S.</dc:creator>
<dc:creator>Dueker, K.</dc:creator>
<dc:creator>Vo, A. L.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Capozzola, T.</dc:creator>
<dc:creator>Joyce, C.</dc:creator>
<dc:creator>Zhao, F.</dc:creator>
<dc:creator>Anzanello, F.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Bibollet-Ruche, F.</dc:creator>
<dc:creator>Ramos, A.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Lewis, M. G.</dc:creator>
<dc:creator>Ozorowski, G.</dc:creator>
<dc:creator>Landais, E.</dc:creator>
<dc:creator>Foley, B. T.</dc:creator>
<dc:creator>Wagh, K.</dc:creator>
<dc:creator>Sok, D.</dc:creator>
<dc:creator>Briney, B.</dc:creator>
<dc:creator>Ward, A. B.</dc:creator>
<dc:creator>Hahn, B. H.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Shaw, G. M.</dc:creator>
<dc:creator>Andrabi, R.</dc:creator>
<dc:date>2023-09-23</dc:date>
<dc:identifier>doi:10.1101/2023.09.21.558743</dc:identifier>
<dc:title><![CDATA[Germline-targeting chimpanzee SIV Envelopes induce V2-apex broadly neutralizing-like B cell precursors in a rhesus macaque infection model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.21.558870v1?rss=1">
<title>
<![CDATA[
S100A8/A9 predicts triple-negative breast cancer response to PIM kinase and PD-1/PD-L1 inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.21.558870v1?rss=1</link>
<description><![CDATA[
It remains elusive why some triple-negative breast cancer (TNBC) patients respond poorly to existing therapies while others respond well. Our retrospective analysis of historical gene expression datasets reveals that increased expression of immunosuppressive cytokine S100A8/A9 in early-stage tumors is robustly associated with subsequent disease progression in TNBC. Although it has recently gained recognition as a potential anticancer target, S100A8/A9 has not been integrated into clinical study designs evaluating molecularly targeted therapies. Our small molecule screen has identified PIM kinase inhibitors as capable of decreasing S100A8/A9 expression in multiple cell types, including TNBC and immunosuppressive myeloid cells. Furthermore, combining PIM inhibition and immune checkpoint blockade induces significant antitumor responses, especially in otherwise resistant S100A8/A9-high PD-1/PD-L1-positive tumors. Importantly, serum S100A8/A9 levels mirror those of tumor S100A8/A9 in a syngeneic mouse model of TNBC. Thus, our data suggest that S100A8/A9 could be a predictive and pharmacodynamic biomarker in clinical trials evaluating combination therapy targeting PIM and immune checkpoints in TNBC and encourage the development of S100A8/A9-based liquid biopsy tests.
]]></description>
<dc:creator>Begg, L. R.</dc:creator>
<dc:creator>Orriols, A. M.</dc:creator>
<dc:creator>Zannikou, M.</dc:creator>
<dc:creator>Yeh, C.</dc:creator>
<dc:creator>Vadlamani, P.</dc:creator>
<dc:creator>Kanojia, D.</dc:creator>
<dc:creator>Bolin, R.</dc:creator>
<dc:creator>Dunne, S. F.</dc:creator>
<dc:creator>Balakrishnan, S.</dc:creator>
<dc:creator>Camarda, R.</dc:creator>
<dc:creator>Roth, D.</dc:creator>
<dc:creator>Zielinski-Mozny, N. A.</dc:creator>
<dc:creator>Yau, C.</dc:creator>
<dc:creator>Vassilopoulos, A.</dc:creator>
<dc:creator>Huang, T.-H.</dc:creator>
<dc:creator>Kim, K.-Y. A.</dc:creator>
<dc:creator>Horiuchi, D.</dc:creator>
<dc:date>2023-09-23</dc:date>
<dc:identifier>doi:10.1101/2023.09.21.558870</dc:identifier>
<dc:title><![CDATA[S100A8/A9 predicts triple-negative breast cancer response to PIM kinase and PD-1/PD-L1 inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.22.558764v1?rss=1">
<title>
<![CDATA[
Predicting Long and Short Duration Beta Bursts from Subthalamic Nucleus Local Field Potential Activity in Parkinson's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.22.558764v1?rss=1</link>
<description><![CDATA[
Neural activities within the beta frequency range (13-30 Hz) are not stationary, but occur in transient packets known as beta bursts. Parkinsons disease (PD) is characterized by the occurrence of beta bursts of increased duration and amplitude within the cortico-basal ganglia network. The pathophysiological importance of beta bursts is exemplified by the fact that they serve as a clinically useful feedback signal in beta amplitude triggered adaptive Deep Brain Stimulation (aDBS). Prolonged duration beta bursts are closely associated with motor impairments in PD, whilst bursts of shorter duration may have a physiological role. Consequently, we aimed to develop a deep learning-based pipeline capable of predicting long (> 150ms) and short (< 150ms) duration beta bursts from subthalamic nucleus local field potential (LFP) recordings. Our approach achieved promising accuracy values of 87% and 85.2% in two patients implanted with a DBS device that was capable of long-term wireless LFP sensing. Our findings highlight the feasibility of prolonged beta burst prediction and could inform the development of a new type of intelligent DBS approach with the capability of delivering stimulation only during the occurrence of prolonged bursts.
]]></description>
<dc:creator>Abdi-Sargezeh, B.</dc:creator>
<dc:creator>Shirani, S.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Starr, P.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:creator>Oswal, A.</dc:creator>
<dc:date>2023-09-23</dc:date>
<dc:identifier>doi:10.1101/2023.09.22.558764</dc:identifier>
<dc:title><![CDATA[Predicting Long and Short Duration Beta Bursts from Subthalamic Nucleus Local Field Potential Activity in Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.24.559205v1?rss=1">
<title>
<![CDATA[
A developmentally defined population of neurons in the lateral septum controls responses to aversive stimuli 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.24.559205v1?rss=1</link>
<description><![CDATA[
When interacting with their environment, animals must balance exploratory and defensive behavior to evaluate and respond to potential threats. The lateral septum (LS) is a structure in the ventral forebrain that calibrates the magnitude of behavioral responses to stress-related external stimuli, including the regulation of threat avoidance. The complex connectivity between the LS and other parts of the brain, together with its largely unexplored neuronal diversity, makes it difficult to understand how defined LS circuits control specific behaviors. Here, we describe a mouse model in which a population of neurons with a common developmental origin (Nkx2.1-lineage neurons) are absent from the LS. Using a combination of circuit tracing and behavioral analyses, we found that these neurons receive inputs from the perifornical area of the anterior hypothalamus (PeFAH) and are specifically activated in stressful contexts. Mice lacking Nkx2.1-lineage LS neurons display increased exploratory behavior even under stressful conditions. Our study extends the current knowledge about how defined neuronal populations within the LS can evaluate contextual information to select appropriate behavioral responses. This is a necessary step towards understanding the crucial role that the LS plays in neuropsychiatric conditions where defensive behavior is dysregulated, such as anxiety and aggression disorders.
]]></description>
<dc:creator>Turrero Garcia, M.</dc:creator>
<dc:creator>Tran, D. N.</dc:creator>
<dc:creator>Peterson, R. E.</dc:creator>
<dc:creator>Stegmann, S. K.</dc:creator>
<dc:creator>Hanson, S. M.</dc:creator>
<dc:creator>Reid, C. M.</dc:creator>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Vu, S.</dc:creator>
<dc:creator>Harwell, C. C.</dc:creator>
<dc:date>2023-09-25</dc:date>
<dc:identifier>doi:10.1101/2023.09.24.559205</dc:identifier>
<dc:title><![CDATA[A developmentally defined population of neurons in the lateral septum controls responses to aversive stimuli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.26.559634v1?rss=1">
<title>
<![CDATA[
Expansion of interferon signaling-associated gene (ISAG)-hi T cells in early-onset Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.26.559634v1?rss=1</link>
<description><![CDATA[
INTRODUCTIONAltered immune signatures are emerging as a central theme in neurodegenerative disease, yet little is known about immune responses in early-onset Alzheimers disease (EOAD).

METHODSWe examined single-cell RNA-sequencing (scRNA-seq) data from peripheral blood mononuclear cells (PBMCs) and droplet digital (dd)PCR data from CD4 T cells from participants with EOAD and clinically normal controls.

RESULTSWe analyzed ~182,000 PBMCs by scRNA-seq and discovered increased interferon signaling-associated gene (ISAG) expression and striking expansion of antiviral-like ISAGhi T cells in EOAD. We isolated CD4 T cells from additional EOAD cases and confirmed increased expression of ISAGhi marker genes. Publicly available cerebrospinal fluid leukocyte scRNA-seq data from late-onset mild cognitive impairment and AD also revealed increased expression of interferon-response genes.

DISCUSSIONISAGhi T cells, apparently primed for antiviral activity, are expanded in EOAD. Additional research into these cells and the role of heightened peripheral IFN signaling in neurodegeneration is warranted.
]]></description>
<dc:creator>Sirkis, D. W.</dc:creator>
<dc:creator>Solsberg, C. W.</dc:creator>
<dc:creator>Johnson, T. P.</dc:creator>
<dc:creator>Geier, E. G.</dc:creator>
<dc:creator>Bonham, L. W.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Rabinovici, G. D.</dc:creator>
<dc:creator>Yokoyama, J. S.</dc:creator>
<dc:date>2023-09-27</dc:date>
<dc:identifier>doi:10.1101/2023.09.26.559634</dc:identifier>
<dc:title><![CDATA[Expansion of interferon signaling-associated gene (ISAG)-hi T cells in early-onset Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.25.559419v1?rss=1">
<title>
<![CDATA[
Distinct transcriptional profiles of maternal and fetal placental macrophages at term are associated with gravidity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.25.559419v1?rss=1</link>
<description><![CDATA[
Maternal intervillous monocytes (MIMs) and fetal Hofbauer cells (HBCs) are myeloid-derived immune cells at the maternal-fetal interface. Little is known regarding the molecular phenotypes and roles of these distinct monocyte/macrophage populations. Here, we used RNA sequencing to investigate the transcriptional profiles of MIMs and HBCs in six normal term pregnancies. Our analyses revealed distinct transcriptomes of MIMs and HBCs. Genes involved in differentiation and cell organization pathways were more highly expressed in MIMs vs. HBCs. In contrast, HBCs had higher expression of genes involved in inflammatory responses and cell surface receptor signaling. Maternal gravidity influenced monocyte programming, as expression of pro-inflammatory molecules was significantly higher in MIMs from multigravidas compared to primigravidas. In HBCs, multigravidas displayed enrichment of gene pathways involved in cell-cell signaling and differentiation. In summary, our results demonstrated that MIMs and HBCs have highly divergent transcriptional signatures, reflecting their distinct origins, locations, functions, and roles in inflammatory responses. Our data further suggested that maternal gravidity influences the gene signatures of MIMs and HBCs, potentially modulating the interplay between tolerance and trained immunity. The phenomenon of reproductive immune memory may play a novel role in the differential susceptibility of primigravidas to pregnancy complications.
]]></description>
<dc:creator>Ozarslan, N.</dc:creator>
<dc:creator>Robinson, J. F.</dc:creator>
<dc:creator>Buarpung, S.</dc:creator>
<dc:creator>Kim, M. Y.</dc:creator>
<dc:creator>Ansbro, M. R.</dc:creator>
<dc:creator>Akram, J.</dc:creator>
<dc:creator>Montoya, D. J.</dc:creator>
<dc:creator>Kamya, M. R.</dc:creator>
<dc:creator>Kakuru, A.</dc:creator>
<dc:creator>Dorsey, G.</dc:creator>
<dc:creator>Rosenthal, P. J.</dc:creator>
<dc:creator>Cheng, G.</dc:creator>
<dc:creator>Feeney, M. E.</dc:creator>
<dc:creator>Fisher, S. J.</dc:creator>
<dc:creator>Gaw, S. L.</dc:creator>
<dc:date>2023-09-28</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.559419</dc:identifier>
<dc:title><![CDATA[Distinct transcriptional profiles of maternal and fetal placental macrophages at term are associated with gravidity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.26.559470v1?rss=1">
<title>
<![CDATA[
Multimodal identification of rare potent effector CD8 T cells in solid tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.26.559470v1?rss=1</link>
<description><![CDATA[
The anti-tumor function of CD8 T cells is limited through well-established pathways of T cell exhaustion (TEX). Strategies to capture emergent functional states amongst this dominant trajectory of dysfunction are necessary to find pathways to durable anti-tumor immunity. By leveraging transcriptional reporting (by the fluorescent protein TFP) of the T cell activation marker Cd69, related to upstream AP-1 transcription factors, we define a classifier for potent versus sub-optimal CD69+ activation states arising from T cell stimulation. In tumors, this delineation acts an additional functional readout along the TEX differentiation trajectory, within and across TEX subsets, marked by enhanced effector cytokine and granzyme B production. The more potent state remains differentially prominent in a T cell-mediated tumor clearance model, where they also show increased engagement in the microenvironment and are superior in tumor cell killing. Employing multimodal CITE-Seq in human head and neck tumors enables a similar strategy to identify Cd69RNAhiCD69+ cells that also have enhanced functional features in comparison to Cd69RNAloCD69+ cells, again within and across intratumoral CD8 T cell subsets. Refining the contours of the T cell functional landscape in tumors in this way paves the way for the identification of rare exceptional effectors, with imminent relevance to cancer treatment.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=120 SRC="FIGDIR/small/559470v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Ray, A.</dc:creator>
<dc:creator>Bassette, M.</dc:creator>
<dc:creator>Hu, K. H.</dc:creator>
<dc:creator>Pass, L. F.</dc:creator>
<dc:creator>Samad, B.</dc:creator>
<dc:creator>Combes, A.</dc:creator>
<dc:creator>Johri, V.</dc:creator>
<dc:creator>Davidson, B.</dc:creator>
<dc:creator>Hernandez, G.</dc:creator>
<dc:creator>Zaleta-Linares, I.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2023-09-28</dc:date>
<dc:identifier>doi:10.1101/2023.09.26.559470</dc:identifier>
<dc:title><![CDATA[Multimodal identification of rare potent effector CD8 T cells in solid tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.27.559406v1?rss=1">
<title>
<![CDATA[
HybriSeq: Probe-based Device-free Single-cell RNA Profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.27.559406v1?rss=1</link>
<description><![CDATA[
We have developed the HybriSeq method for single-cell RNA profiling, which utilizes in situ hybridization of multiple probes for targeted transcripts, followed by split-pool barcoding and sequencing analysis of the probes. We have shown that HybriSeq can achieve high sensitivity for RNA detection with multiple probes and profile entire transcripts without an end bias. The utility of HybriSeq is demonstrated in characterizing cell-to-cell heterogeneities of a panel of 196 genes in peripheral blood mononuclear cells and the detection of missed annotations of transcripts.
]]></description>
<dc:creator>Foyt, D.</dc:creator>
<dc:creator>Brown, D.</dc:creator>
<dc:creator>Zhou, S.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:date>2023-09-29</dc:date>
<dc:identifier>doi:10.1101/2023.09.27.559406</dc:identifier>
<dc:title><![CDATA[HybriSeq: Probe-based Device-free Single-cell RNA Profiling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.27.559768v1?rss=1">
<title>
<![CDATA[
Best practices for perturbation MPRA--a computational evaluation framework of sequence design strategies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.27.559768v1?rss=1</link>
<description><![CDATA[
The advent of the perturbation-based massively parallel reporter assays (MPRAs) technique has enabled delineating of the roles of non-coding regulatory elements in orchestrating gene expression. However, computational efforts remain scant to evaluate and establish guidelines for sequence design strategies for perturbation MPRAs. Here, we propose a framework for evaluating and comparing various perturbation strategies for MPRA experiments. Under this framework, we benchmark three different perturbation approaches from the perspectives of alteration in motif-based profiles, consistency of MPRA outputs, and robustness of models that predict the activities of putative regulatory motifs. Although our analyses show similar while significant results in multiple metrics, the method of randomly shuffling nucleotides outperform the other two methods. Thus, we still recommend designing sequences by randomly shuffling the nucleotides of the perturbed site in perturbation-MPRA. The evaluation framework, together with the benchmarking findings in our work, creates a resource of computational pipelines and illustrates the promise of perturbation-MPRA for predicting non-coding regulatory activities.
]]></description>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Ashuach, T.</dc:creator>
<dc:creator>Inoue, F.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Yosef, N.</dc:creator>
<dc:creator>Kreimer, A.</dc:creator>
<dc:date>2023-09-29</dc:date>
<dc:identifier>doi:10.1101/2023.09.27.559768</dc:identifier>
<dc:title><![CDATA[Best practices for perturbation MPRA--a computational evaluation framework of sequence design strategies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.29.560231v1?rss=1">
<title>
<![CDATA[
Adolescent oligodendrogenesis and myelination restrict experience-dependent neuronal plasticity in adult visual cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.29.560231v1?rss=1</link>
<description><![CDATA[
BACKGROUNDDevelopmental myelination is a protracted process in the mammalian brain. One theory for why oligodendrocytes mature so slowly posits that myelination may stabilize neuronal circuits and temper neuronal plasticity as animals age. We tested this hypothesis in the visual cortex, which has a well-defined critical period for experience-dependent neuronal plasticity.

OBJECTIVES/METHODSTo prevent myelin progression, we conditionally deleted Myrf, a transcription factor necessary for oligodendrocyte maturation, from oligodendrocyte precursor cells (Myrf cKO) in adolescent mice. To induce experience-dependent plasticity, adult control and Myrf cKO mice were monocularly deprived by eyelid suture. Functional and structural neuronal plasticity in the visual cortex were assessed in vivo by intrinsic signal optical imaging and longitudinal two photon imaging of dendritic spines, respectively.

RESULTSDuring adolescence, visual experience modulated the rate of oligodendrocyte maturation in visual cortex. Myrf deletion from oligodendrocyte precursors during adolescence led to inhibition of oligodendrocyte maturation and myelination that persisted into adulthood. Following monocular deprivation, visual cortex activity in response to visual stimulation of the deprived eye remained stable in adult control mice, as expected for post-critical period animals. By contrast, visual cortex responses to the deprived eye decreased significantly following monocular deprivation in adult Myrf cKO mice, reminiscent of the plasticity observed in adolescent mice. Furthermore, visual cortex neurons in adult Myrf cKO mice had fewer dendritic spines and a higher level of spine turnover. Finally, monocular deprivation induced spatially coordinated spine size decreases in adult Myrf cKO, but not control, mice.

CONCLUSIONSThese results demonstrate a critical role for oligodendrocytes in shaping the maturation and stabilization of cortical circuits and support the concept of myelin acting as a brake on neuronal plasticity during development.
]]></description>
<dc:creator>Xin, W.</dc:creator>
<dc:creator>Kaneko, M.</dc:creator>
<dc:creator>Roth, R. H.</dc:creator>
<dc:creator>Zhang, A.</dc:creator>
<dc:creator>Nocera, S.</dc:creator>
<dc:creator>Ding, J. B.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:creator>Chan, J. R.</dc:creator>
<dc:date>2023-09-30</dc:date>
<dc:identifier>doi:10.1101/2023.09.29.560231</dc:identifier>
<dc:title><![CDATA[Adolescent oligodendrogenesis and myelination restrict experience-dependent neuronal plasticity in adult visual cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.29.560238v1?rss=1">
<title>
<![CDATA[
BiP/GRP78 is a pro-viral factor for diverse dsDNA viruses that promotes the survival and proliferation of cells upon KSHV infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.29.560238v1?rss=1</link>
<description><![CDATA[
The Endoplasmic Reticulum (ER)-resident HSP70 chaperone BiP (HSPA5) plays a crucial role in maintaining and restoring protein folding homeostasis in the ER. BiPs function is often dysregulated in cancer and virus-infected cells, conferring pro-oncogenic and pro-viral advantages. We explored BiPs functions during infection by the Kaposis sarcoma-associated herpesvirus (KSHV), an oncogenic gamma-herpesvirus associated with cancers of immunocompromised patients. Our findings reveal that BiP protein levels are upregulated in infected epithelial cells during the lytic phase of KSHV infection. This upregulation occurs independently of the unfolded protein response (UPR), a major signaling pathway that regulates BiP availability. Genetic and pharmacological inhibition of BiP halts KSHV viral replication and reduces the proliferation and survival of KSHV-infected cells. Notably, inhibition of BiP limits the spread of other alpha- and beta-herpesviruses and poxviruses with minimal toxicity for normal cells. Our work suggests that BiP is a potential target for developing broad-spectrum antiviral therapies against double-stranded DNA viruses and a promising candidate for therapeutic intervention in KSHV-related malignancies.
]]></description>
<dc:creator>Najarro, G.</dc:creator>
<dc:creator>Brackett, K.</dc:creator>
<dc:creator>Woosley, H.</dc:creator>
<dc:creator>Faeldonea, C.</dc:creator>
<dc:creator>Moreno, O. K.</dc:creator>
<dc:creator>Ramirez Negron, A.</dc:creator>
<dc:creator>Love, C.</dc:creator>
<dc:creator>Ward, R.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>Gardner, B. M.</dc:creator>
<dc:creator>Arias, C.</dc:creator>
<dc:date>2023-09-30</dc:date>
<dc:identifier>doi:10.1101/2023.09.29.560238</dc:identifier>
<dc:title><![CDATA[BiP/GRP78 is a pro-viral factor for diverse dsDNA viruses that promotes the survival and proliferation of cells upon KSHV infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.29.560245v1?rss=1">
<title>
<![CDATA[
Disease-specific selective vulnerability and neuroimmune pathways in dementia revealed by single cellgenomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.29.560245v1?rss=1</link>
<description><![CDATA[
Summary/AbstractThe development of successful therapeutics for dementias requires an understanding of their shared and distinct molecular features in the human brain. We performed single-nuclear RNAseq and ATACseq in Alzheimer disease (AD), Frontotemporal degeneration (FTD), and Progressive Supranuclear Palsy (PSP), analyzing 40 participants, yielding over 1.4M cells from three brain regions ranging in vulnerability and pathological burden. We identify 35 shared disease-associated cell types and 14 that are disease-specific, replicating those previously identified in AD. Disease-specific cell states represent molecular features of disease-specific glial-immune mechanisms and neuronal vulnerability in each disorder, layer 4/5 intra-telencephalic neurons in AD, layer 2/3 intra-telencephalic neurons in FTD, and layer 5/6 near-projection neurons in PSP. We infer intrinsic disease-associated gene regulatory networks, which we empirically validate by chromatin footprinting. We find that causal genetic risk acts in specific neuronal and glial cells that differ across disorders, primarily non-neuronal cells in AD and specific neuronal subtypes in FTD and PSP. These data illustrate the heterogeneous spectrum of glial and neuronal composition and gene expression alterations in different dementias and identify new therapeutic targets by revealing shared and disease-specific cell states.
]]></description>
<dc:creator>Rexach, J. E.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Polioudakis, D.</dc:creator>
<dc:creator>Lin, L.-C.</dc:creator>
<dc:creator>Mitri, V.</dc:creator>
<dc:creator>Elkins, A.</dc:creator>
<dc:creator>Yin, A.</dc:creator>
<dc:creator>Calini, D.</dc:creator>
<dc:creator>Kawaguchi, R.</dc:creator>
<dc:creator>Ou, J.</dc:creator>
<dc:creator>Huang, J.</dc:creator>
<dc:creator>Williams, C.</dc:creator>
<dc:creator>Robinson, J.</dc:creator>
<dc:creator>Gaus, S.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Lee, E.</dc:creator>
<dc:creator>Grinberg, L.</dc:creator>
<dc:creator>Vinters, H.</dc:creator>
<dc:creator>Trojanowski, J.</dc:creator>
<dc:creator>Seeley, W.</dc:creator>
<dc:creator>Molhatra, D.</dc:creator>
<dc:creator>Geschwind, D.</dc:creator>
<dc:date>2023-09-30</dc:date>
<dc:identifier>doi:10.1101/2023.09.29.560245</dc:identifier>
<dc:title><![CDATA[Disease-specific selective vulnerability and neuroimmune pathways in dementia revealed by single cellgenomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.30.560321v1?rss=1">
<title>
<![CDATA[
Transcriptional coupling of telomeric retrotransposons with the cell cycle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.30.560321v1?rss=1</link>
<description><![CDATA[
Instead of employing telomerases to safeguard chromosome ends, dipteran species maintain their telomeres by transposition of telomeric-specific retrotransposons (TRs): in Drosophila, these are HeT-A, TART, and TAHRE. Previous studies have shown how these TRs create tandem repeats at chromosome ends, but the exact mechanism controlling TR transcription has remained unclear. Here we report the identification of multiple subunits of the transcription cofactor Mediator complex and transcriptional factors Scalloped (Sd, the TEAD homolog in flies) and E2F1-Dp as novel regulators of TR transcription and telomere length in Drosophila. Depletion of multiple Mediator subunits, Dp, or Sd increased TR expression and telomere length, while over-expressing E2F1-Dp or knocking down the E2F1 regulator Rbf1 (Retinoblastoma-family protein 1) stimulated TR transcription, with Mediator and Sd affecting TR expression through E2F1-Dp. The CUT&RUN analysis revealed direct binding of CDK8, Dp, and Sd to telomeric repeats. These findings highlight the essential role of the Mediator complex in maintaining telomere homeostasis by regulating TR transcription through E2F1-Dp and Sd, revealing the intricate coupling of TR transcription with the host cell-cycle machinery, thereby ensuring chromosome end protection and genomic stability during cell division.
]]></description>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Xie, X.-J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Sun, J.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Hemba-Waduge, R.- U.-S.</dc:creator>
<dc:creator>Ji, J.-y.</dc:creator>
<dc:date>2023-10-01</dc:date>
<dc:identifier>doi:10.1101/2023.09.30.560321</dc:identifier>
<dc:title><![CDATA[Transcriptional coupling of telomeric retrotransposons with the cell cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.29.560039v1?rss=1">
<title>
<![CDATA[
Commensal myeloid crosstalk in neonatal skin regulates long-term cutaneous type 17 inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.29.560039v1?rss=1</link>
<description><![CDATA[
Early life microbe-immune interactions at barrier surfaces have lasting impacts on the trajectory towards health versus disease. Monocytes, macrophages and dendritic cells are primary sentinels in barrier tissues, yet the salient contributions of commensal-myeloid crosstalk during tissue development remain poorly understood. Here, we identify that commensal microbes facilitate accumulation of a population of monocytes in neonatal skin. Transient postnatal depletion of these monocytes resulted in heightened IL-17A production by skin T cells, which was particularly sustained among CD4+ T cells and sufficient to exacerbate inflammatory skin pathologies. Neonatal skin monocytes were enriched in expression of negative regulators of the IL-1 pathway. Functional in vivo experiments confirmed a key role for excessive IL-1R1 signaling in T cells as contributing to the dysregulated type 17 response in neonatal monocyte-depleted mice. Thus, a commensal-driven wave of monocytes into neonatal skin critically facilitates immune homeostasis in this prominent barrier tissue.
]]></description>
<dc:creator>Dhariwala, M. O.</dc:creator>
<dc:creator>DeRogatis, A. M.</dc:creator>
<dc:creator>Okoro, J. N.</dc:creator>
<dc:creator>Weckel, A. A.</dc:creator>
<dc:creator>Tran, V. M.</dc:creator>
<dc:creator>Habrylo, I.</dc:creator>
<dc:creator>Ojewumi, O. T.</dc:creator>
<dc:creator>Tammen, A. E.</dc:creator>
<dc:creator>Leech, J. M.</dc:creator>
<dc:creator>Merana, G. R.</dc:creator>
<dc:creator>Carale, R. O.</dc:creator>
<dc:creator>Barrere-Cain, R.</dc:creator>
<dc:creator>Hiam-Galvez, K. J.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:creator>Scharschmidt, T. C.</dc:creator>
<dc:date>2023-10-02</dc:date>
<dc:identifier>doi:10.1101/2023.09.29.560039</dc:identifier>
<dc:title><![CDATA[Commensal myeloid crosstalk in neonatal skin regulates long-term cutaneous type 17 inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.02.560519v1?rss=1">
<title>
<![CDATA[
Lysosomal proteomics reveals mechanisms of neuronal apoE4-associated lysosomal dysfunction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.02.560519v1?rss=1</link>
<description><![CDATA[
ApoE4 is the primary risk factor for Alzheimer Disease (AD). Early AD pathological events first affect the neuronal endolysosomal system, which in turn causes neuronal protein aggregation and cell death. Despite the crucial influence of lysosomes upon AD pathophysiology, and that apoE4 localizes to lysosomes, the influence of apoE4 on lysosomal function remains unexplored. We find that expression of apoE4 in neuronal cell lines results in lysosomal alkalinization and impaired lysosomal function. To identify driving factors for these defects, we performed quantitative lysosomal proteome profiling. This revealed that apoE4 expression results in differential regulation of numerous lysosomal proteins, correlating with apoE allele status and disease severity in AD brains. In particular, apoE4 expression results in the depletion of lysosomal Lgals3bp and the accumulation of lysosomal Tmed5. We additionally validated that these lysosomal protein changes can be targeted to modulate lysosomal function. Taken together, this work thereby reveals that apoE4 causes widespread lysosomal defects through remodeling the lysosomal proteome, with the lysosomal Tmed5 accumulation and Lgals3bp depletion manifesting as lysosomal alkalinization in apoE4 neurons.
]]></description>
<dc:creator>Krogsaeter, E. K.</dc:creator>
<dc:creator>McKetney, J.</dc:creator>
<dc:creator>Marquez, A.</dc:creator>
<dc:creator>Cakir, Z.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Jang, G. M.</dc:creator>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Zhou, A.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:date>2023-10-02</dc:date>
<dc:identifier>doi:10.1101/2023.10.02.560519</dc:identifier>
<dc:title><![CDATA[Lysosomal proteomics reveals mechanisms of neuronal apoE4-associated lysosomal dysfunction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.02.560569v1?rss=1">
<title>
<![CDATA[
BNT162b2 mRNA vaccine-induced sex differences in the single-cell transcriptome of peripheral blood mononuclear cells in healthy adults 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.02.560569v1?rss=1</link>
<description><![CDATA[
IntroductionMen reportedly experience more severe disease and adverse outcomes from COVID-19, including death. Women report more adverse events (AEs) after vaccination in general. While few studies have addressed sex-specific risk factors or molecular mechanisms behind COVID-19, none have examined sex differences in the response to COVID-19 vaccination.

MethodsWe searched AE reporting databases to find sex differences specific to COVID-19 vaccines. We analyzed public datasets to identify baseline sex differences in gene expression across cell types and time points, and sex differences in the response to the second BNT162b2 mRNA vaccine dose.

ResultsSex differences in AE rates for mRNA vaccines equaled those for other non-mRNA vaccines. T cells and monocytes showed the greatest number of sexually dimorphic genes. Platelet counts in the study population differed significantly before vaccination (3.6% in females vs 1.8% in males) but not after the second BNT162b2 dose (7.2% vs 7.3%). There were no notable sex differences in the expression of key genes induced by the second dose after exclusion of platelets. BNT162b2 dose 2-specific APOBEC3Ahigh monocytes and the dose 2-induced gene signature persisted for longer in women. Glucocorticoid-responsive TSC22D3, CEBPB/D and DDIT4 were specifically induced in females; the voltage-gated potassium channel regulatory subunit KCNE3 was specifically induced in males.

ConclusionsThis sexual dimorphism in both X-linked and autosomal gene transcriptome in PBMCs after mRNA COVID-19 vaccination might explain fatigue, autoimmune, and neurological AEs reported after vaccination at different rates in women and men.
]]></description>
<dc:creator>Knapp, J. D.</dc:creator>
<dc:creator>Bhargava, A.</dc:creator>
<dc:date>2023-10-03</dc:date>
<dc:identifier>doi:10.1101/2023.10.02.560569</dc:identifier>
<dc:title><![CDATA[BNT162b2 mRNA vaccine-induced sex differences in the single-cell transcriptome of peripheral blood mononuclear cells in healthy adults]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.03.560675v1?rss=1">
<title>
<![CDATA[
Targeting the Hippo pathway in cancers via ubiquitination dependent TEAD degradation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.03.560675v1?rss=1</link>
<description><![CDATA[
The Hippo pathway is among the most frequently altered key signaling pathways in cancer. TEAD1-4 are essential transcription factors and key downstream effectors in the Hippo pathway in human cells. Here, we identified RNF146 as a ubiquitin ligase (E3) of TEADs, which negatively regulates their stability in cells through proteasome-mediated degradation. We show that RNF146-mediated TEAD ubiquitination is dependent on the TEAD PARylation state. We further validated the genetic interaction between RNF146 and the Hippo pathway in cancer cell lines and the model organism Drosophila melanogaster. Despite the RNF146 and proteasome-mediated degradation mechanisms, TEADs are stable proteins with a long half-life in cells. We demonstrate that degradation of TEADs can be greatly enhanced pharmacologically with heterobifunctional chemical inducers of protein degradation (CIDEs). These TEAD-CIDEs can effectively suppress activation of YAP/TAZ target genes in a dose-dependent manner and exhibit significant anti-proliferative effects in YAP/TAZ-dependent tumor cells, thus phenocopying the effect of genetic ablation of TEAD protein. Collectively, this study demonstrates that the ubiquitin-proteasome system plays an important role in regulating TEAD functions and provides a proof-of-concept demonstration that pharmacologically induced TEAD ubiquitination could be leveraged to target YAP/TAZ-driven cancers.
]]></description>
<dc:creator>Pham, T. H.</dc:creator>
<dc:creator>Pahuja, K. B.</dc:creator>
<dc:creator>Hagenbeek, T. J.</dc:creator>
<dc:creator>Zbieg, J.</dc:creator>
<dc:creator>Noland, C. L.</dc:creator>
<dc:creator>Pham, V. C.</dc:creator>
<dc:creator>Yao, X.</dc:creator>
<dc:creator>Rose, C. M.</dc:creator>
<dc:creator>Browder, K. C.</dc:creator>
<dc:creator>Lee, H.-J.</dc:creator>
<dc:creator>Yu, M.</dc:creator>
<dc:creator>Liang-Chu, M.</dc:creator>
<dc:creator>Martin, S. E.</dc:creator>
<dc:creator>Verschueren, E.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Kubala, M. H.</dc:creator>
<dc:creator>Fong, R.</dc:creator>
<dc:creator>Lorenzo, M.</dc:creator>
<dc:creator>Beroza, P.</dc:creator>
<dc:creator>Hsu, P.</dc:creator>
<dc:creator>Paul, S.</dc:creator>
<dc:creator>Villemure, E.</dc:creator>
<dc:creator>Lee, W.</dc:creator>
<dc:creator>Cheung, T.</dc:creator>
<dc:creator>Clausen, S.</dc:creator>
<dc:creator>Lacap, J.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Schmidt, S.</dc:creator>
<dc:creator>Modrusan, Z.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:creator>Crawford, J.</dc:creator>
<dc:creator>Jasper, H.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Lill, J. R.</dc:creator>
<dc:creator>Malek, S.</dc:creator>
<dc:creator>Rudolph, J.</dc:creator>
<dc:creator>Wertz, I. E.</dc:creator>
<dc:creator>Chang, M.</dc:creator>
<dc:creator>Ye, X.</dc:creator>
<dc:creator>Dey, A.</dc:creator>
<dc:date>2023-10-04</dc:date>
<dc:identifier>doi:10.1101/2023.10.03.560675</dc:identifier>
<dc:title><![CDATA[Targeting the Hippo pathway in cancers via ubiquitination dependent TEAD degradation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.03.560703v1?rss=1">
<title>
<![CDATA[
Palatal segment contributions to midfacial growth in three inbred mouse strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.03.560703v1?rss=1</link>
<description><![CDATA[
Anterior-posterior (A-P) elongation of the palate is a critical aspect of integrated midfacial morphogenesis. Reciprocal epithelial-mesenchymal interactions drive secondary palate elongation that is coupled to the periodic formation of signaling centers within the rugae growth zone (RGZ). However, the relationship between RGZ-driven morphogenetic processes, the differentiative dynamics of underlying palatal bone mesenchymal precursors, and the segmental organization of the upper jaw has remained enigmatic. A detailed ontogenetic study of these relationships is important because palatal segment growth is a critical aspect of normal midfacial growth, can produce dysmorphology when altered, and is a likely basis for evolutionary differences in upper jaw morphology. We completed a combined whole mount gene expression and morphometric analysis of normal murine palatal segment growth dynamics and resulting upper jaw morphology. Our results demonstrated that the first formed palatal ruga (ruga 1), found just posterior to the RGZ, maintained an association with important nasal, neurovascular and palatal structures throughout early midfacial development. This suggested that these features are positioned at a proximal source of embryonic midfacial directional growth. Our detailed characterization of midfacial morphogenesis revealed a one-to-one relationship between palatal segments and upper jaw bones during the earliest stages of palatal elongation. Growth of the maxillary anlage within the anterior secondary palate is uniquely coupled to RGZ-driven morphogenesis. This may help drive the unequaled proportional elongation of the anterior secondary palate segment prior to palatal shelf fusion. Our results also demonstrated that the future maxillary-palatine suture, approximated by the position of ruga 1 and consistently associated with the palatine anlage, formed predominantly via the posterior differentiation of the maxilla within the expanding anterior secondary palate. Our ontogenetic analysis provides a novel and detailed picture of the earliest spatiotemporal dynamics of intramembranous midfacial skeletal specification and differentiation within the context of the surrounding palatal segment A-P elongation and associated rugae formation.
]]></description>
<dc:creator>Welsh, I. C.</dc:creator>
<dc:creator>Feiler, M.</dc:creator>
<dc:creator>Lipman, D.</dc:creator>
<dc:creator>Mormile, I.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>Percival, C. J. C.</dc:creator>
<dc:date>2023-10-05</dc:date>
<dc:identifier>doi:10.1101/2023.10.03.560703</dc:identifier>
<dc:title><![CDATA[Palatal segment contributions to midfacial growth in three inbred mouse strains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.03.560769v1?rss=1">
<title>
<![CDATA[
MOIRE: A software package for the estimation of allele frequenciesand effective multiplicity of infection from polyallelic data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.03.560769v1?rss=1</link>
<description><![CDATA[
MotivationMalaria parasite genetic data can provide insight into parasite phenotypes, evolution, and transmission. However, estimating key parameters such as allele frequencies, multiplicity of infection (MOI), and within-host relatedness from genetic data is challenging, particularly in the presence of multiple related coinfecting strains. Existing methods often rely on single nucleotide polymorphism (SNP) data and do not account for within-host relatedness.

ResultsWe present MOIRE (Multiplicity Of Infection and allele frequency REcovery), a Bayesian approach to estimate allele frequencies, MOI, and within-host relatedness from genetic data subject to experimental error. MOIRE accommodates both polyallelic and SNP data, making it applicable to diverse genotyping panels. We also introduce a novel metric, the effective MOI (eMOI), which integrates MOI and within-host relatedness, providing a robust and interpretable measure of genetic diversity. Extensive simulations and real-world data from a malaria study in Namibia demonstrate the superior performance of MOIRE over naive estimation methods, accurately estimating MOI up to 7 with moderate sized panels of diverse loci (e.g. microhaplotypes). MOIRE also revealed substantial heterogeneity in population mean MOI and mean relatedness across health districts in Namibia, suggesting detectable differences in transmission dynamics. Notably, eMOI emerges as a portable metric of within-host diversity, facilitating meaningful comparisons across settings when allele frequencies or genotyping panels differ. Compared to existing software, MOIRE enables more comprehensive insights into within-host diversity and population structure.

AvailabilityMOIRE is available as an R package at https://eppicenter.github.io/moire/.

Contactmm@maxmurphy.dev

Supplementary informationSupplementary data are available at Bioinformatics online.
]]></description>
<dc:creator>Murphy, M.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:date>2023-10-05</dc:date>
<dc:identifier>doi:10.1101/2023.10.03.560769</dc:identifier>
<dc:title><![CDATA[MOIRE: A software package for the estimation of allele frequenciesand effective multiplicity of infection from polyallelic data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.04.560905v1?rss=1">
<title>
<![CDATA[
Introduction of Dicistrovirus IRESs into UAS/SV40-polyA constructs results in premature polyadenylation and strong overexpression of the upstream ORF in Drosophila animals. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.04.560905v1?rss=1</link>
<description><![CDATA[
To evaluate the properties of insect virus internal ribosomal entry sites (IRESs) for protein expression in Drosophila, we have introduced Cricket Paralysis virus (CrPV) and Drosophila C virus (DCV) IRESs into UAS/SV40-polyA vector. We found that introduction of IRESs induce premature polyadenylation, resulting in both truncation of the mRNA, and an increase in mRNA levels of approximately 40-fold. The increase in mRNA levels was accompanied by increased resistance to nonsense-mediated mRNA decay (NMD)-mediated degradation. Our results suggest that premature polyadenylation increases mRNA stability in the SV40 polyadenylation site-containing constructs, suggesting a novel method for robust overexpression of transgenes in Drosophila.
]]></description>
<dc:creator>Lidsky, P. V.</dc:creator>
<dc:creator>Dmitriev, S. E.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:date>2023-10-05</dc:date>
<dc:identifier>doi:10.1101/2023.10.04.560905</dc:identifier>
<dc:title><![CDATA[Introduction of Dicistrovirus IRESs into UAS/SV40-polyA constructs results in premature polyadenylation and strong overexpression of the upstream ORF in Drosophila animals.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.05.561059v1?rss=1">
<title>
<![CDATA[
Structural and molecular basis of choline uptake into the brain by FLVCR2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.05.561059v1?rss=1</link>
<description><![CDATA[
Choline is an essential nutrient that the human body needs in vast quantities for cell membrane synthesis, epigenetic modification, and neurotransmission. The brain has a particularly high demand for choline, but how it enters the brain has eluded the field for over fifty years. The MFS transporter FLVCR1 was recently determined to be a choline transporter, and while this protein is not highly expressed at the blood-brain barrier (BBB), its relative FLVCR2 is. Previous studies have shown that mutations in human Flvcr2 cause cerebral vascular abnormalities, hydrocephalus, and embryonic lethality, but the physiological role of FLVCR2 is unknown. Here, we demonstrate both in vivo and in vitro that FLVCR2 is a BBB choline transporter and is responsible for the majority of choline uptake into the brain. We also determine the structures of choline-bound FLVCR2 in the inward- and outward-facing states using cryo-electron microscopy to 2.49 and 2.77 [A] resolution, respectively. These results reveal how the brain obtains choline and provide molecular-level insights into how FLVCR2 binds choline in an aromatic cage and mediates its uptake. Our work could provide a novel framework for the targeted delivery of neurotherapeutics into the brain.
]]></description>
<dc:creator>Cater, R. J.</dc:creator>
<dc:creator>Mukherjee, D.</dc:creator>
<dc:creator>Gil Iturbe, E.</dc:creator>
<dc:creator>Erramilli, S. K.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Koo, K.</dc:creator>
<dc:creator>Santander, N. G.</dc:creator>
<dc:creator>Reckers, A. R.</dc:creator>
<dc:creator>Kloss, B.</dc:creator>
<dc:creator>Gawda, T.</dc:creator>
<dc:creator>Choy, B. C.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Clarke, O. B.</dc:creator>
<dc:creator>Yee, S. W.</dc:creator>
<dc:creator>Kossiakoff, A. K.</dc:creator>
<dc:creator>Quick, M.</dc:creator>
<dc:creator>Arnold, T.</dc:creator>
<dc:creator>Mancia, F.</dc:creator>
<dc:date>2023-10-05</dc:date>
<dc:identifier>doi:10.1101/2023.10.05.561059</dc:identifier>
<dc:title><![CDATA[Structural and molecular basis of choline uptake into the brain by FLVCR2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.04.559423v1?rss=1">
<title>
<![CDATA[
Cross-Tissue Specificity of Pediatric DNA Methylation Associated with Cumulative Family Adversity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.04.559423v1?rss=1</link>
<description><![CDATA[
BackgroundCumulative family adversity (cumulative FA), characterized by co-occurring stressors in a family context, may be biologically embedded through DNA methylation (DNAm) and contribute to later health outcomes.

Materials & MethodsWe compared epigenome-wide DNAm associated with cumulative FA in buccal epithelial cells (BECs; n=218) and peripheral blood mononuclear cells (PBMCs; n=51) from 7-13-year-old children in Canada, accounting for sex, age, predicted cell-type proportion, and genetic ancestry.

ResultsHigher levels of cumulative FA were associated with DNAm at seven sites, primarily in stress- and immune-related genes, only in PBMCs. Negative mother-child interaction contributed to this association.

ConclusionsThe findings of this study suggested that PBMC DNAm can be used as a marker for biological embedding of cumulative FA.
]]></description>
<dc:creator>Chan, M. H.-M.</dc:creator>
<dc:creator>Merrill, S. M.</dc:creator>
<dc:creator>Fatima, F.</dc:creator>
<dc:creator>MacIsaac, J. L.</dc:creator>
<dc:creator>Obradovic, J.</dc:creator>
<dc:creator>Boyce, W. T.</dc:creator>
<dc:creator>Kobor, M. S.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.04.559423</dc:identifier>
<dc:title><![CDATA[Cross-Tissue Specificity of Pediatric DNA Methylation Associated with Cumulative Family Adversity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.04.560781v1?rss=1">
<title>
<![CDATA[
Sexual Differences In The Interaction Between Alcohol And mTORC1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.04.560781v1?rss=1</link>
<description><![CDATA[
The kinase mechanistic target of rapamycin complex 1 (mTORC1) plays an essential role in learning and memory by promoting mRNA to protein translation of a subset of synaptic proteins at dendrites. We generated a large body of data in male rodents indicating that mTORC1 is critically involved in mechanisms that promote numerous adverse behaviors associated with alcohol use disorder (AUD) including heavy alcohol use. For example, we found that mTORC1 is activated in the nucleus accumbens (NAc) and orbitofrontal cortex (OFC) of male mice and rats that were subjected to 7 weeks of intermittent access to 20% alcohol two-bottle choice (IA20%2BC). We further showed that systemic or intra-NAc administration of the selective mTORC1 inhibitor, rapamycin, decreases alcohol seeking and drinking, whereas intra-OFC administration of rapamycin reduces alcohol seeking and habit in male rats. This study aimed to assess mTORC1 activation in these corticostriatal regions of female mice and to determine whether the selective mTORC1 inhibitor, rapamycin, can be used to reduce heavy alcohol use in female mice. We found that mTORC1 is not activated by 7 weeks of intermittent 20% alcohol binge drinking and withdrawal in the NAc and OFC. Like in males, mTORC1 signaling was not activated by chronic alcohol intake and withdrawal in the medial prefrontal cortex (mPFC) of female mice. Interestingly, Pearson correlation comparisons revealed that the basal level of mTORC1 activation between the two prefrontal regions, OFC and mPFC were correlated and that the drinking profile predicts the level of mTORC1 activation in the mPFC after 4-hour binge drinking. Finally, we report that administration of rapamycin does not attenuate heavy alcohol drinking in female animals. Together, our results suggest a sex-dependent contribution of mTORC1 to the neuroadaptation that drives alcohol use and abuse.
]]></description>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>Phamluong, K.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.04.560781</dc:identifier>
<dc:title><![CDATA[Sexual Differences In The Interaction Between Alcohol And mTORC1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.05.560352v1?rss=1">
<title>
<![CDATA[
Deciphering Cell-types and Gene Signatures Associated with Disease Activity in Rheumatoid Arthritis using Single Cell RNA-sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.05.560352v1?rss=1</link>
<description><![CDATA[
ObjectiveSingle cell profiling of synovial tissue has previously identified gene signatures associated with rheumatoid arthritis (RA) pathophysiology, but synovial tissue is difficult to obtain. This study leverages single cell sequencing of peripheral blood mononuclear cells (PBMCs) from patients with RA and matched healthy controls to identify disease relevant cell subsets and cell type specific signatures of disease.

MethodsSingle-cell RNA sequencing (scRNAseq) was performed on peripheral blood mononuclear cells (PBMCs) from 18 RA patients and 18 matched controls, accounting for age, gender, race, and ethnicity). Samples were processed using standard CellRanger and Scanpy pipelines, pseudobulk differential gene expression analysis was performed using DESeq2, and cell-cell communication analysis using CellChat.

ResultsWe identified 18 distinct PBMC subsets, including a novel IFITM3+ monocyte subset. CD4+ T effector memory cells were increased in patients with moderate to high disease activity (DAS28-CRP [&ge;] 3.2), while non-classical monocytes were decreased in patients with low disease activity or remission (DAS28-CRP < 3.2). Differential gene expression analysis identified RA-associated genes in IFITM3+ and non-classical monocyte subsets, and downregulation of pro-inflammatory genes in the V{delta} subset. Additionally, we identified gene signatures associated with disease activity, characterized by upregulation of pro-inflammatory genes TNF, JUN, EGR1, IFIT2, MAFB, G0S2, and downregulation of HLA-DQB1, HLA-DRB5, TNFSF13B. Notably, cell-cell communication analysis revealed upregulation of immune-associated signaling pathways, including VISTA, in patients with RA.

ConclusionsWe provide a novel single-cell transcriptomics dataset of PBMCs from patients with RA, and identify insights into the systemic cellular and molecular mechanisms underlying RA disease activity.
]]></description>
<dc:creator>Binvignat, M.</dc:creator>
<dc:creator>Miao, B.</dc:creator>
<dc:creator>Wibrand, C.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Rychkov, D.</dc:creator>
<dc:creator>Flynn, E.</dc:creator>
<dc:creator>Nititham, J.</dc:creator>
<dc:creator>Tamaki, W.</dc:creator>
<dc:creator>Khan, U.</dc:creator>
<dc:creator>Carvidi, A.</dc:creator>
<dc:creator>Krueger, M.</dc:creator>
<dc:creator>Niemi, E.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Sellam, J.</dc:creator>
<dc:creator>Mariotti-Ferrandiz, E.</dc:creator>
<dc:creator>Klatzmann, D.</dc:creator>
<dc:creator>Gross, A.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Butte, A. J.</dc:creator>
<dc:creator>Criswell, L.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.05.560352</dc:identifier>
<dc:title><![CDATA[Deciphering Cell-types and Gene Signatures Associated with Disease Activity in Rheumatoid Arthritis using Single Cell RNA-sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.05.561076v1?rss=1">
<title>
<![CDATA[
Variants in the DDX6-CXCR5 autoimmune disease risk locus influence the regulatory network in immune cells and salivary gland 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.05.561076v1?rss=1</link>
<description><![CDATA[
Fine mapping and bioinformatic analysis of the DDX6-CXCR5 genetic risk association in Sjogrens Disease (SjD) and Systemic Lupus Erythematosus (SLE) identified five common SNPs with functional evidence in immune cell types: rs4938573, rs57494551, rs4938572, rs4936443, rs7117261. Functional interrogation of nuclear protein binding affinity, enhancer/promoter regulatory activity, and chromatin-chromatin interactions in immune, salivary gland epithelial, and kidney epithelial cells revealed cell type-specific allelic effects for all five SNPs that expanded regulation beyond effects on DDX6 and CXCR5 expression. Mapping the local chromatin regulatory network revealed several additional genes of interest, including lnc-PHLDB1-1. Collectively, functional characterization implicated the risk alleles of these SNPs as modulators of promoter and/or enhancer activities that regulate cell type-specific expression of DDX6, CXCR5, and lnc-PHLDB1-1, among others. Further, these findings emphasize the importance of exploring the functional significance of SNPs in the context of complex chromatin architecture in disease-relevant cell types and tissues.
]]></description>
<dc:creator>Wiley, M. M.</dc:creator>
<dc:creator>Khatri, B.</dc:creator>
<dc:creator>Joachims, M. L.</dc:creator>
<dc:creator>Tessneer, K. L.</dc:creator>
<dc:creator>Stolarczyk, A. M.</dc:creator>
<dc:creator>Rasmussen, A.</dc:creator>
<dc:creator>Anaya, J.-M.</dc:creator>
<dc:creator>Aqrawi, L. A.</dc:creator>
<dc:creator>Bae, S.-C.</dc:creator>
<dc:creator>Baecklund, E.</dc:creator>
<dc:creator>Bjork, A.</dc:creator>
<dc:creator>Brun, J. G.</dc:creator>
<dc:creator>Bucher, S. M.</dc:creator>
<dc:creator>Dand, N.</dc:creator>
<dc:creator>Eloranta, M.-L.</dc:creator>
<dc:creator>Engelke, F.</dc:creator>
<dc:creator>Forsblad-d'Elia, H.</dc:creator>
<dc:creator>Fugmann, C.</dc:creator>
<dc:creator>Glenn, S. B.</dc:creator>
<dc:creator>Gong, C.</dc:creator>
<dc:creator>Gottenberg, J.-E.</dc:creator>
<dc:creator>Hammenfors, D.</dc:creator>
<dc:creator>Imgenberg-Kreuz, J.</dc:creator>
<dc:creator>Jensen, J. L.</dc:creator>
<dc:creator>Johnsen, S. J. A.</dc:creator>
<dc:creator>Jonsson, M. V.</dc:creator>
<dc:creator>Kelly, J. A.</dc:creator>
<dc:creator>Khanam, S.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Kvarnstrom, M.</dc:creator>
<dc:creator>Mandl, T.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Morris, D. L.</dc:creator>
<dc:creator>Nocturne, G.</dc:creator>
<dc:creator>Norheim, K. B.</dc:creator>
<dc:creator>Olsson, P.</dc:creator>
<dc:creator>Palm, O.</dc:creator>
<dc:creator>Pers, J.-O.</dc:creator>
<dc:creator>Rhodus, N. L.</dc:creator>
<dc:creator>Sjowall, C.</dc:creator>
<dc:creator>Skarstein, K.</dc:creator>
<dc:creator>Taylor, K.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.05.561076</dc:identifier>
<dc:title><![CDATA[Variants in the DDX6-CXCR5 autoimmune disease risk locus influence the regulatory network in immune cells and salivary gland]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.05.561114v1?rss=1">
<title>
<![CDATA[
Differential Requirement for Dimerization of the Membrane-Binding PH-TH Module of BTK in B cells and T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.05.561114v1?rss=1</link>
<description><![CDATA[
Brutons tyrosine kinase (BTK) is a major drug target in immune cells. The membrane-binding pleckstrin-homology and tec-homology (PH-TH) domains of BTK are required for signaling. In vitro, dimerization of the PH-TH module strongly stimulates BTK kinase activity. Whether BTK dimerizes in cells via the PH-TH module, and whether this dimerization is necessary for signaling, is unknown. To address this question, we developed high-throughput mutagenesis assays for BTK function in B cells and T cells. We measured the fitness costs for thousands of point mutations in the PH-TH module and kinase domain, allowing us to assess whether dimerization of the PH-TH module and BTK kinase activity are necessary for function. In Ramos B cells we find that neither PH-TH dimerization nor kinase activity is required for BTK signaling. Instead, in Ramos cells, BTK signaling is enhanced by mutations in the PH-TH module that increase membrane adsorption, even at the cost of reduced PH-TH dimerization. In contrast, in Jurkat T cells, we find that BTK signaling depends on both PH-TH dimerization and kinase activity. Evolutionary analysis shows that BTK proteins in fish and lower organisms, like all Tec kinases other than BTK, lack PH-TH dimerization but have active kinase domains. Thus, PH-TH dimerization is not intrinsically required for Tec-kinase activity, and is a special feature that evolved to exert stricter regulatory control on BTK kinase activity as adaptive immune systems gained increased complexity.
]]></description>
<dc:creator>Eisen, T. J.</dc:creator>
<dc:creator>Ghaffari-Kashani, S.</dc:creator>
<dc:creator>Weiss, A.</dc:creator>
<dc:creator>Kuriyan, J.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.05.561114</dc:identifier>
<dc:title><![CDATA[Differential Requirement for Dimerization of the Membrane-Binding PH-TH Module of BTK in B cells and T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.06.560931v1?rss=1">
<title>
<![CDATA[
Identification of secretory autophagy as a novel mechanism modulating activity-induced synaptic remodeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.06.560931v1?rss=1</link>
<description><![CDATA[
The ability of neurons to rapidly remodel their synaptic structure and strength in response to neuronal activity is highly conserved across species and crucial for complex brain functions. However, mechanisms required to elicit and coordinate the acute, activity-dependent structural changes across synapses are not well understood. Here, using an RNAi screen in Drosophila against genes affecting nervous system functions in humans, we uncouple cellular processes important for synaptic plasticity from synapse development. We find mutations associated with neurodegenerative and mental health disorders are 2-times more likely to affect activity-induced synaptic remodeling than synapse development. We further demonstrate that neuronal activity stimulates autophagy activation but diminishes degradative autophagy, thereby driving the pathway towards autophagy-based secretion. Presynaptic knockdown of Snap29, Sec22, or Rab8, proteins implicated in the secretory autophagy pathway, is sufficient to abolish activity-induced synaptic remodeling. This study uncovers secretory autophagy as a novel trans-synaptic signaling mechanism modulating structural plasticity.
]]></description>
<dc:creator>Chang, Y.-C.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Lee, J. Y.</dc:creator>
<dc:creator>Langen, J.</dc:creator>
<dc:creator>Chang, K. T.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.06.560931</dc:identifier>
<dc:title><![CDATA[Identification of secretory autophagy as a novel mechanism modulating activity-induced synaptic remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.06.561161v1?rss=1">
<title>
<![CDATA[
Structural dynamics of the active HER4 and HER2/HER4 complexes is finely tuned by different growth factors and glycosylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.06.561161v1?rss=1</link>
<description><![CDATA[
Human Epidermal growth factor Receptor 4 (HER4 or ERBB4) carries out essential functions in the development and maintenance of the cardiovascular and nervous systems. HER4 activation is regulated by a diverse group of extracellular ligands including the neuregulin (NRG) family and betacellulin (BTC), which promote HER4 homodimerization or heterodimerization with other HER receptors. Important cardiovascular functions of HER4 are exerted via heterodimerization with its close homolog and orphan receptor, HER2. To date structural insights into ligand-mediated HER4 activation have been limited to crystallographic studies of HER4 ectodomain homodimers in complex with NRG1y. Here we report cryo-EM structures of near full-length HER2/HER4 heterodimers and full-length HER4 homodimers bound to NRG1y and BTC. We show that the structures of the heterodimers bound to either ligand are nearly identical and that in both cases the HER2/HER4 heterodimer interface is less dynamic than those observed in structures of HER2/EGFR and HER2/HER3 heterodimers. In contrast, structures of full-length HER4 homodimers bound to NRG1y and BTC display more large-scale dynamics mirroring states previously reported for EGFR homodimers. Our structures also reveal the presence of multiple glycan modifications within HER4 ectodomains, modeled for the first time in HER receptors, that distinctively contribute to the stabilization of HER4 homodimer interfaces over those of HER2/HER4 heterodimers.
]]></description>
<dc:creator>Trenker, R.</dc:creator>
<dc:creator>Diwanji, D.</dc:creator>
<dc:creator>Bingham, T.</dc:creator>
<dc:creator>Verba, K. A.</dc:creator>
<dc:creator>Jura, N.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.06.561161</dc:identifier>
<dc:title><![CDATA[Structural dynamics of the active HER4 and HER2/HER4 complexes is finely tuned by different growth factors and glycosylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.05.561066v1?rss=1">
<title>
<![CDATA[
CELL-E 2: Translating Proteins to Pictures and Back with a Bidirectional Text-to-Image Transformer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.05.561066v1?rss=1</link>
<description><![CDATA[
We present CELL-E 2, a novel bidirectional transformer that can generate images depicting protein subcellular localization from the amino acid sequences (and vice versa). Protein localization is a challenging problem that requires integrating sequence and image information, which most existing methods ignore. CELL-E 2 extends the work of CELL-E, not only capturing the spatial complexity of protein localization and produce probability estimates of localization atop a nucleus image, but also being able to generate sequences from images, enabling de novo protein design. We train and finetune CELL-E 2 on two large-scale datasets of human proteins. We also demonstrate how to use CELL-E 2 to create hundreds of novel nuclear localization signals (NLS). Results and interactive demos are featured at https://bohuanglab.github.io/CELL-E_2/.
]]></description>
<dc:creator>Khwaja, E.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Agarunov, A.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:date>2023-10-07</dc:date>
<dc:identifier>doi:10.1101/2023.10.05.561066</dc:identifier>
<dc:title><![CDATA[CELL-E 2: Translating Proteins to Pictures and Back with a Bidirectional Text-to-Image Transformer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.06.561183v1?rss=1">
<title>
<![CDATA[
The Hierarchy of Psychedelic Effects: Three Systematic Reviews and Meta-Analyses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.06.561183v1?rss=1</link>
<description><![CDATA[
Serotonergic psychedelics induce altered states of consciousness and have shown potential for treating a variety of neuropsychiatric disorders, including depression and addiction. Yet their modes of action are not fully understood. Here, we provide a novel, synergistic understanding of psychedelics arising from systematic reviews and meta-analyses of three hierarchical levels of analysis: 1) subjective experience (phenomenology), 2) neuroimaging and 3) molecular pharmacology. Phenomenologically, medium and high doses of LSD yield significantly higher ratings of visionary restructuralisation than psilocybin on the 5-dimensional Altered States of Consciousness Scale. Our neuroimaging results reveal that, in general, psychedelics significantly strengthen between-network functional connectivity (FC) while significantly diminishing within-network FC. Pharmacologically, LSD induces significantly more inositol phosphate formation at the 5-HT2A receptor than DMT and psilocin, yet there are no significant between-drug differences in the selectivity of psychedelics for the 5-HT2A, 5-HT2C, or D2 receptors, relative to the 5-HT1A receptor. Our meta-analyses link DMT, LSD, and psilocybin to specific neural fingerprints at each level of analysis. The results show a highly non-linear relationship between these fingerprints. Overall, our analysis highlighted the high heterogeneity and risk of bias in the literature. This suggests an urgent need for standardising experimental procedures and analysis techniques, as well as for more research on the emergence between different levels of psychedelic effects.
]]></description>
<dc:creator>Shinozuka, K.</dc:creator>
<dc:creator>Jerotic, K.</dc:creator>
<dc:creator>Mediano, P. A. M.</dc:creator>
<dc:creator>Zhao, A. T.</dc:creator>
<dc:creator>Preller, K. H.</dc:creator>
<dc:creator>Carhart-Harris, R. L.</dc:creator>
<dc:creator>Kringelbach, M. L.</dc:creator>
<dc:date>2023-10-07</dc:date>
<dc:identifier>doi:10.1101/2023.10.06.561183</dc:identifier>
<dc:title><![CDATA[The Hierarchy of Psychedelic Effects: Three Systematic Reviews and Meta-Analyses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.08.561433v1?rss=1">
<title>
<![CDATA[
Proteome birthdating reveals age-selectivity of protein ubiquitination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.08.561433v1?rss=1</link>
<description><![CDATA[
Within a cell, proteins have distinct and highly variable half-lives. As a result, the molecular ages of proteins can range from seconds to years. How the age of a protein influences its environmental interactions is a largely unexplored area of biology. To investigate the age-selectivity of cellular pathways, we developed a methodology termed "proteome birthdating" that barcodes proteins based on their time of synthesis. We demonstrate that this approach provides accurate measurements of protein turnover kinetics without the requirement for multiple kinetic time points. As a first use case of the birthdated proteome, we investigated the age distribution of the human ubiquitinome. Our results indicate that the vast majority of ubiquitinated proteins in a cell consist of newly synthesized proteins and that these young proteins constitute the bulk of the degradative flux through the proteasome. Rapidly ubiquitinated nascent proteins are enriched in cytosolic subunits of large protein complexes. Conversely, proteins destined for the secretory pathway and vesicular transport have older ubiquitinated populations. Our data also identified a smaller subset of very old ubiquitinated cellular proteins that do not appear to be targeted to the proteasome for rapid degradation. Together, our data provide an age census of the human ubiquitinome and establish proteome birthdating as a robust methodology for investigating the protein age-selectivity of diverse cellular pathways.

Significance StatementCellular proteins have widely different ages - whereas some have been recently synthesized, others have existed in the cell for days or even years. How a proteins age influences its functions and interactions is largely unknown because it is difficult to globally differentiate proteins based on their time of synthesis. To address this challenge, we developed an analytical method named "proteome birthdating" that can partition cellular proteins into multiple discernible age groups. As an example application, we used proteome birthdating to examine the protein age-selectivity of the ubiquitin proteasome system, a major protein degradation pathway in eukaryotes. Our results show that proteins destined for degradation by this pathway consist of either particularly young or particularly old proteins, with the former being the predominant population. Together, our results establish proteome birthdating as a useful approach for analyzing the turnover of proteins and investigating the functional consequences of their age.
]]></description>
<dc:creator>Meadow, M. E.</dc:creator>
<dc:creator>Broas, S.</dc:creator>
<dc:creator>Hoare, M.</dc:creator>
<dc:creator>Alimohammadi, F.</dc:creator>
<dc:creator>Welle, K. A.</dc:creator>
<dc:creator>Swovick, K.</dc:creator>
<dc:creator>Hryhorenko, J. R.</dc:creator>
<dc:creator>Martinez, J. C.</dc:creator>
<dc:creator>Biashad, S. A.</dc:creator>
<dc:creator>Seluanov, A.</dc:creator>
<dc:creator>Gorbunova, V.</dc:creator>
<dc:creator>Buchwalter, A.</dc:creator>
<dc:creator>Ghaemmaghami, S.</dc:creator>
<dc:date>2023-10-08</dc:date>
<dc:identifier>doi:10.1101/2023.10.08.561433</dc:identifier>
<dc:title><![CDATA[Proteome birthdating reveals age-selectivity of protein ubiquitination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.07.561344v1?rss=1">
<title>
<![CDATA[
Exploring the tumor micro-environment in ovarian cancer histotypes and tumor sites 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.07.561344v1?rss=1</link>
<description><![CDATA[
Ovarian cancer is a highly heterogeneous disease consisting of at least five different histological subtypes with varying clinical features, cells of origin, molecular composition, risk factors, and treatments. While most single-cell studies have focused on High grade serous ovarian cancer, a comprehensive landscape of the constituent cell types and their interactions within the tumor microenvironment are yet to be established in the different ovarian cancer histotypes. Further characterization of tumor progression, metastasis, and various histotypes are also needed to connect molecular signatures to pathological grading for personalized diagnosis and tailored treatment. In this study, we leveraged high-resolution single-cell RNA sequencing technology to elucidate the cellular compositions on 21 solid tumor samples collected from 12 patients with six ovarian cancer histotypes and both primary (ovaries) and metastatic (omentum, rectum) sites. The diverse collection allowed us to deconstruct the histotypes and tumor site-specific expression patterns of cells in the tumor and identify key marker genes and ligand-receptor pairs that are active in the ovarian tumor microenvironment. Our findings can be used in improving precision disease stratification and optimizing treatment options.
]]></description>
<dc:creator>Xie, B.</dc:creator>
<dc:creator>Olalekan, S.</dc:creator>
<dc:creator>BACK, R.</dc:creator>
<dc:creator>Ashitey, N. A. A. A.</dc:creator>
<dc:creator>Basu, A.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.07.561344</dc:identifier>
<dc:title><![CDATA[Exploring the tumor micro-environment in ovarian cancer histotypes and tumor sites]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.08.561428v1?rss=1">
<title>
<![CDATA[
Developmental origin and local signals cooperate to determine septal astrocyte identity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.08.561428v1?rss=1</link>
<description><![CDATA[
Astrocyte specification during development is influenced by both intrinsic and extrinsic factors, but the precise contribution of each remains poorly understood. Here we show that septal astrocytes from Nkx2.1 and Zic4 expressing progenitor zones are allocated into non-overlapping domains of the medial (MS) and lateral septal nuclei (LS) respectively. Astrocytes in these areas exhibit distinctive molecular and morphological features tailored to the unique cellular and synaptic circuit environment of each nucleus. Using single-nucleus (sn) RNA sequencing, we trace the developmental trajectories of cells in the septum and find that neurons and astrocytes undergo region and developmental stage-specific local cell-cell interactions. We show that expression of the classic morphogens Sonic hedgehog (Shh) and Fibroblast growth factors (Fgfs) by MS and LS neurons respectively, functions to promote the molecular specification of local astrocytes in each region. Finally, using heterotopic cell transplantation, we show that both morphological and molecular specifications of septal astrocytes are highly dependent on the local microenvironment, regardless of developmental origins. Our data highlights the complex interplay between intrinsic and extrinsic factors shaping astrocyte identities and illustrates the importance of the local environment in determining astrocyte functional specialization.
]]></description>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Reid, C. M.</dc:creator>
<dc:creator>Granados, A. A.</dc:creator>
<dc:creator>Garcia, M. T.</dc:creator>
<dc:creator>Dale-Huang, F.</dc:creator>
<dc:creator>Hanson, S. M.</dc:creator>
<dc:creator>Leon, W. R. M.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Adam, M. A.</dc:creator>
<dc:creator>Mosto, O.</dc:creator>
<dc:creator>Pisco, A. O.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:creator>Harwell, C. C.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.08.561428</dc:identifier>
<dc:title><![CDATA[Developmental origin and local signals cooperate to determine septal astrocyte identity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.09.561578v1?rss=1">
<title>
<![CDATA[
A feasibility study of smartphone sensors to assess the effect of acute high altitude (3,800 m) exposure on cognition and motor function in healthy participants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.09.561578v1?rss=1</link>
<description><![CDATA[
Acute exposure to hypoxia at attitude has neurologic effects. Some subjects develop severe neurologic symptoms, including Parkinsonism, when hypoxic at high altitude as part of an Acute Mountain Sickness syndrome. Digital health technologies can provide continuous monitoring and objective, real-world measures of movement disorders, but only limited validation data of wearable sensors is available in the high-altitude setting. This equipment validation and feasibility study assessed smartphone microphone and accelerometer function at sea level and 12470 feet (3,800 m) to assess their potential value to predict and prevent neurologic features of acute mountain sickness. A feasibility study of standardized assessments of motor, speech and cognitive tasks was performed in 3 normal subjects at sea level and at altitude. All subjects were hypoxic at altitude with O2 saturations ranging from 77-81%. Shaker table (range of frequencies) and high-fidelity speaker (range of frequencies) controls confirmed high correlation of observed and expected measurements for microphone and accelerometer under all conditions. The feasibility study demonstrated that under conditions of hypoxia at attitude, fine motor skills are impaired; visual short-term memory is not impaired but has longer response time; gait and balance is impaired, and a postural tremor develops with frequencies below 10 Hz. Future studies could use these wearable sensors to further assess effects at altitude of more severe hypoxia with applications in the high-altitude environment for Parkinsons Disease patients, with further opportunity for aviation and military use.
]]></description>
<dc:creator>Goldman, O. G.</dc:creator>
<dc:creator>Dubowitz, G.</dc:creator>
<dc:creator>Anderson, D.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.09.561578</dc:identifier>
<dc:title><![CDATA[A feasibility study of smartphone sensors to assess the effect of acute high altitude (3,800 m) exposure on cognition and motor function in healthy participants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.10.561642v1?rss=1">
<title>
<![CDATA[
Convergent Epigenetic Evolution Drives Relapse in Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.10.561642v1?rss=1</link>
<description><![CDATA[
Relapse of acute myeloid leukemia (AML) is highly aggressive and often treatment refractory. We analyzed previously published AML relapse cohorts and found that 40% of relapses occur without changes in driver mutations, suggesting that non-genetic mechanisms drive relapse in a large proportion of cases. We therefore characterized epigenetic patterns of AML relapse using 26 matched diagnosis-relapse samples with ATAC-seq. This analysis identified a relapse-specific chromatin accessibility signature for mutationally stable AML, suggesting that AML undergoes epigenetic evolution at relapse independent of mutational changes. Analysis of leukemia stem cell (LSC) chromatin changes at relapse indicated that this leukemic compartment underwent significantly less epigenetic evolution than non-LSCs, while epigenetic changes in non-LSCs reflected overall evolution of the bulk leukemia. Finally, we used single-cell ATAC-seq paired with mitochondrial sequencing (mtscATAC) to map clones from diagnosis into relapse along with their epigenetic features. We found that distinct mitochondrially-defined clones exhibit more similar chromatin accessibility at relapse relative to diagnosis, demonstrating convergent epigenetic evolution in relapsed AML. These results demonstrate that epigenetic evolution is a feature of relapsed AML and that convergent epigenetic evolution can occur following treatment with induction chemotherapy.
]]></description>
<dc:creator>Nuno, K.</dc:creator>
<dc:creator>Azizi, A.</dc:creator>
<dc:creator>Koehnke, T.</dc:creator>
<dc:creator>Lareau, C.</dc:creator>
<dc:creator>Ediriwickrema, A.</dc:creator>
<dc:creator>Corces, M. R.</dc:creator>
<dc:creator>Satpathy, A. T.</dc:creator>
<dc:creator>Majeti, R.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.10.561642</dc:identifier>
<dc:title><![CDATA[Convergent Epigenetic Evolution Drives Relapse in Acute Myeloid Leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.10.561687v1?rss=1">
<title>
<![CDATA[
Interplay of RAP2 GTPase and the cytoskeleton in Hippo pathway regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.10.561687v1?rss=1</link>
<description><![CDATA[
The Hippo signaling is instrumental in regulating organ size, regeneration, and carcinogenesis. The cytoskeleton emerges as a primary Hippo signaling modulator. Its structural alterations in response to environmental and intrinsic stimuli control Hippo kinase cascade activity. However, the precise mechanisms underlying the cytoskeleton regulation of Hippo signaling are not fully understood. RAP2 GTPase is known to mediate the mechanoresponses of Hippo signaling via activating the core Hippo kinases LATS1/2 through MAP4Ks and MST1/2. Here we show the pivotal role of the reciprocal regulation between RAP2 GTPase and the cytoskeleton in Hippo signaling. RAP2 deletion undermines the responses of the Hippo pathway to external cues tied to RhoA GTPase inhibition and actin cytoskeleton remodeling, such as energy stress and serum deprivtion. Notably, RhoA inhibitors and actin disruptors fail to activate LATS1/2 effectively in RAP2-deficient cells. RNA sequencing highlighted differential regulation of both actin and microtubule networks by RAP2 gene deletion. Consistently, Taxol, a microtubule-stabilizing agent, was less effective in activating LATS1/2 and inhibiting cell growth in RAP2 and MAP4K4/6/7 knockout cells. In summary, our findings position RAP2 as a central integrator of cytoskeletal signals for Hippo signaling, which offers new avenues for understanding Hippo regulation and therapeutic interventions in Hippo-impaired cancers.
]]></description>
<dc:creator>Wu, C.</dc:creator>
<dc:creator>Cai, X.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Rodriguez, C. D.</dc:creator>
<dc:creator>Herrmann, L.</dc:creator>
<dc:creator>Zoaldi, G.</dc:creator>
<dc:creator>Huang, C.-Y.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Sanghvi, V. R.</dc:creator>
<dc:creator>Lu, R. O.</dc:creator>
<dc:creator>Meng, Z.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.10.561687</dc:identifier>
<dc:title><![CDATA[Interplay of RAP2 GTPase and the cytoskeleton in Hippo pathway regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.10.561706v1?rss=1">
<title>
<![CDATA[
Control of intestinal stemness and cell lineage by histone variant H2A.Z isoforms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.10.561706v1?rss=1</link>
<description><![CDATA[
The histone variant H2A.Z plays important functions in the regulation of gene expression. In mammals, it is encoded by two genes, giving raise to two highly related isoforms named H2A.Z.1 and H2A.Z.2, which can have similar or antagonistic functions depending on the promoter. Knowledge of the physiopathological consequences of such functions emerges, but how the balance between these isoforms regulates tissue homeostasis is not fully understood. Here, we investigated the relative role of H2A.Z isoforms in intestinal epithelial homeostasis. Through genome-wide analysis of H2A.Z genomic localization in differentiating Caco-2 cells, we uncovered an enrichment of H2A.Z isoforms on the bodies of genes which are induced during enterocyte differentiation, stressing the potential importance of H2A.Z isoforms dynamics in this process. Through a combination of in vitro and in vivo experiments, we further demonstrated the two isoforms cooperate for stem and progenitor cells proliferation, as well as for secretory lineage differentiation. However, we found that they antagonistically regulate enterocyte differentiation, with H2A.Z.1 preventing terminal differentiation and H2A.Z.2 favoring it. Altogether, these data indicate that H2A.Z isoforms are critical regulators of intestine homeostasis and may provide a paradigm of how the balance between two isoforms of the same chromatin structural protein can control physiopathological processes.
]]></description>
<dc:creator>Rispal, J.</dc:creator>
<dc:creator>Rives, C.</dc:creator>
<dc:creator>Jouffret, V.</dc:creator>
<dc:creator>Leoni, C.</dc:creator>
<dc:creator>Dubois, L.</dc:creator>
<dc:creator>Chevillard-Briet, M.</dc:creator>
<dc:creator>Trouche, D.</dc:creator>
<dc:creator>Escaffit, F.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.10.561706</dc:identifier>
<dc:title><![CDATA[Control of intestinal stemness and cell lineage by histone variant H2A.Z isoforms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.10.561722v1?rss=1">
<title>
<![CDATA[
Neurogenesis-mediated circuit remodeling reduces engram reinstatement and promotes forgetting 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.10.561722v1?rss=1</link>
<description><![CDATA[
Post-training increases in hippocampal neurogenesis are associated with forgetting of hippocampus-dependent memories in adult mice. This form of forgetting might be due to increased numbers of new neurons, remodeling of hippocampal circuitry or some combination of both. Here we tested the hypothesis that neurogenesis-mediated forgetting is caused by remodeling of hippocampal circuits by engineering mice in which adult-generated granule cells hypo- or hyper-integrate into hippocampal circuits. Using gene deletion, opto- and chemogenetic strategies, we find that hypo-integration of newborn neurons prevents post- training exercise-induced forgetting of contextual fear memories. Conversely, inducing hyper- integration of newborn neurons following contextual fear conditioning is sufficient to produce forgetting. Because these interventions did not affect survival of newborn neurons, these findings suggest that neurogenesis-mediated remodeling of hippocampal circuits represents a continuous and active form of interference that alters accessibility of engrams underlying hippocampal memories. Consistent with this, using engram-labeling approaches, we found that exercise-induced forgetting was associated with reduced engram reactivation.
]]></description>
<dc:creator>Guskjolen, A.</dc:creator>
<dc:creator>Dhaliwal, J.</dc:creator>
<dc:creator>de la Parra, J.</dc:creator>
<dc:creator>Epp, J. R.</dc:creator>
<dc:creator>Ko, S.</dc:creator>
<dc:creator>Chahley, E.</dc:creator>
<dc:creator>de Snoo, M.</dc:creator>
<dc:creator>Solari, M.</dc:creator>
<dc:creator>Panesar, P.</dc:creator>
<dc:creator>Snyder, J. S.</dc:creator>
<dc:creator>Duan, X.</dc:creator>
<dc:creator>Sanes, J.</dc:creator>
<dc:creator>Josselyn, S.</dc:creator>
<dc:creator>Frankland, P.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.10.561722</dc:identifier>
<dc:title><![CDATA[Neurogenesis-mediated circuit remodeling reduces engram reinstatement and promotes forgetting]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.09.561582v1?rss=1">
<title>
<![CDATA[
The single-molecule accessibility landscape of newly replicated mammalian chromatin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.09.561582v1?rss=1</link>
<description><![CDATA[
The higher-order structure of newly replicated (i.e.  nascent) chromatin fibers remains poorly-resolved, limiting our understanding of how epigenomes are maintained across cell divisions. To address this, we present Replication-Aware Single-molecule Accessibility Mapping (RASAM), a long-read sequencing method that nondestructively measures genome-wide replication-status and protein-DNA interactions simultaneously on intact chromatin templates. We report that individual human and mouse nascent chromatin fibers are  hyperaccessible compared to steady-state chromatin. This hyperaccessibility occurs at two, coupled length-scales: first, individual nucleosome core particles on nascent DNA exist as a mixture of partially-unwrapped nucleosomes and other subnucleosomal species; second, newly-replicated chromatin fibers are significantly enriched for irregularly-spaced nucleosomes on individual DNA molecules. Focusing on specific cis-regulatory elements (e.g. transcription factor binding sites; active transcription start sites [TSSs]), we discover unique modes by which nascent chromatin hyperaccessibility is resolved at the single-molecule level: at CCCTC-binding factor (CTCF) binding sites, CTCF and nascent nucleosomes compete for motifs on nascent chromatin fibers, resulting in quantitatively-reduced CTCF occupancy and motif accessibility post-replication; at active TSSs, high levels of steady-state chromatin accessibility are preserved, implying that nucleosome free regions (NFRs) are rapidly re-established behind the fork. Our study introduces a new paradigm for studying higher-order chromatin fiber organization behind the replication fork. More broadly, we uncover a unique organization of newly replicated chromatin that must be reset by active processes, providing a substrate for epigenetic reprogramming.
]]></description>
<dc:creator>Ostrowski, M. S.</dc:creator>
<dc:creator>Yang, M. G.</dc:creator>
<dc:creator>McNally, C. P.</dc:creator>
<dc:creator>Abdulhay, N. J.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Nora, E. P.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:date>2023-10-11</dc:date>
<dc:identifier>doi:10.1101/2023.10.09.561582</dc:identifier>
<dc:title><![CDATA[The single-molecule accessibility landscape of newly replicated mammalian chromatin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.11.561685v1?rss=1">
<title>
<![CDATA[
Population genetic analyses of longitudinal vaginal microbiome reveal racioethnic evolutionary dynamics and prevailing positive selection of Lactobacillus adhesins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.11.561685v1?rss=1</link>
<description><![CDATA[
The vaginal microbiome is important for reproductive health and should constantly evolve in response to dynamic host-microbe interactions. The composition of the vaginal microbiome is associated with ethnicity; however, the evolutionary landscape of the vaginal microbiome, especially in the multi-ethnic context, remains under-studied. In this study, we performed a longitudinal evolutionary vaginal microbiome analysis of 351 samples from 35 pregnant women with diverse ethnic backgrounds and validated the main findings in two cohorts totaling 462 samples from 90 multi-ethnic women. Microbiome alpha diversity and community states showed strong ethnic signatures. Lactobacillaceae species had a significantly higher nonsynonymous/synonymous mutation ratio (pN/pS) than non-Lactobacillaceae species in all ethnicities. In addition, non-Lactobacillaceae anaerobic bacteria were enriched in Black and Latino women, with significantly elevated nucleotide diversity and lower pN/pS in Black women. Intriguingly, the Lactobacillaceae species had a large repertoire of positively selected genes, including the human mucin-binding and bacterial cell wall anchor genes, which showed independent, recurrent signatures of positive selection across multiple strains, indicating that the host-microbiome interactions directly drive microbial evolution at the molecular interface. Finally, we propose that the evolutionary metrics reflect the environmental niches of adapting microbes. Our study revealed the extensive ethnic signatures in vaginal microbial diversity, composition, community state, and evolutionary dynamics at species and gene levels, highlighting the importance of studying the host-microbiome ecosystem from an evolutionary perspective.

HighlightsO_LIExtensive ethnic signatures in the vaginal microbial diversity, composition, community state, and evolutionary dynamics were demonstrated.
C_LIO_LIHealthy Lactobacillaceae species showed lower nucleotide diversity but much more relaxed or even positive selection of genes than non-Lactobacillaceae species.
C_LIO_LIIn Black women, non-Lactobacillaceae species displayed higher nucleotide diversity and more stringent negative selection.
C_LIO_LILactobacillus mucin-binding and cell wall anchor genes showed convergent signatures of positive selection across vaginal microbiomes.
C_LI
]]></description>
<dc:creator>Wei, X.</dc:creator>
<dc:creator>Tsai, M.-S.</dc:creator>
<dc:creator>Liang, L.</dc:creator>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>Hung, C.-J.</dc:creator>
<dc:creator>Jelliffe-Pawlowski, L.</dc:creator>
<dc:creator>Rand, L.</dc:creator>
<dc:creator>Snyder, M.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:date>2023-10-11</dc:date>
<dc:identifier>doi:10.1101/2023.10.11.561685</dc:identifier>
<dc:title><![CDATA[Population genetic analyses of longitudinal vaginal microbiome reveal racioethnic evolutionary dynamics and prevailing positive selection of Lactobacillus adhesins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.11.561735v1?rss=1">
<title>
<![CDATA[
A Simple Subcortical Feeding Circuit Linking Interoceptive Inputs to Consummatory Behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.11.561735v1?rss=1</link>
<description><![CDATA[
The brain processes an array of stimuli enabling the selection of an appropriate behavioural response but the neural pathways linking interoceptive inputs to outputs for feeding are poorly understood. Here we delineate a subcortical circuit in which brain-derived neurotrophic factor (BDNF) expressing neurons in the ventromedial hypothalamus (VMH) directly connect interoceptive inputs to motor centers controlling food consumption and jaw movements. VMHBDNF neuron inhibition increases food intake by gating motor sequences of feeding through projections to premotor areas of the jaw. When food is unavailable, VMHBDNF inhibition elicits consummatory behaviors directed at inanimate objects such as a wooden block and inhibition of mesencephalic trigeminal area (Me5) projections evokes rhythmic jaw movements. The activity of these neurons is decreased during food consumption and increases when food is in proximity but not consumed. Activity is also increased in obese animals and after leptin treatment. VMHBDNF neurons receive monosynaptic inputs from both agouti-related peptide (AgRP) and proopiomelanocortin (POMC) neurons in the arcuate nucleus (Arc) and constitutive VMHBDNF activation blocks the orexigenic effect of AgRP activation. These data delineate an Arc[-&gt;]VMHBDNF[-&gt;]Me5 circuit that senses the energy state of an animal and regulates consummatory behaviors in a state dependent manner.
]]></description>
<dc:creator>Kosse, C.</dc:creator>
<dc:creator>Ivanov, J.</dc:creator>
<dc:creator>Knight, Z.</dc:creator>
<dc:creator>Pellegrino, K.</dc:creator>
<dc:creator>Friedman, J.</dc:creator>
<dc:date>2023-10-11</dc:date>
<dc:identifier>doi:10.1101/2023.10.11.561735</dc:identifier>
<dc:title><![CDATA[A Simple Subcortical Feeding Circuit Linking Interoceptive Inputs to Consummatory Behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.11.561932v1?rss=1">
<title>
<![CDATA[
Mapping chromatin state and transcriptional response in CIC-DUX4 undifferentiated round cell sarcoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.11.561932v1?rss=1</link>
<description><![CDATA[
CIC-DUX4 is a rare and understudied transcription factor fusion oncoprotein. CIC-DUX4 co-opts native gene targets to drive a lethal form of human sarcoma. The molecular underpinnings that lead to oncogenic reprograming and CIC-DUX4 sarcomagenesis remain largely undefined. Through an integrative ChIP and RNA-Seq analysis using patient-derived CIC-DUX4 cells, we define CIC-DUX4 mediated chromatin states and function. We show that CIC-DUX4 primarily localizes to proximal and distal cis-regulatory elements where it associates with active histone marks. Our findings nominate key signaling pathways and molecular targets that enable CIC-DUX4 to mediate tumor cell survival. Collectively, our data demonstrate how the CIC-DUX4 fusion oncoprotein impacts chromatin state and transcriptional responses to drive an oncogenic program in undifferentiated sarcoma.

SignificanceCIC-DUX4 sarcoma is a rare and lethal sarcoma that affects children, adolescent young adults, and adults. CIC-DUX4 sarcoma is associated with rapid metastatic dissemination and relative insensitivity to chemotherapy. There are no current standard-of-care therapies for CIC-DUX4 sarcoma leading to universally poor outcomes for patients. Through a deep mechanistic understanding of how the CIC-DUX4 fusion oncoprotein reprograms chromatin state and function, we aim to improve outcomes for CIC-DUX4 patients.
]]></description>
<dc:creator>Thomas, N.</dc:creator>
<dc:creator>Luck, C.</dc:creator>
<dc:creator>Shlimon, N.</dc:creator>
<dc:creator>Kriska Ponce, R.</dc:creator>
<dc:creator>Kosibaty, Z.</dc:creator>
<dc:creator>Okimoto, R.</dc:creator>
<dc:date>2023-10-12</dc:date>
<dc:identifier>doi:10.1101/2023.10.11.561932</dc:identifier>
<dc:title><![CDATA[Mapping chromatin state and transcriptional response in CIC-DUX4 undifferentiated round cell sarcoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.13.562264v1?rss=1">
<title>
<![CDATA[
EEG spectral attractors identify a geometric core of resting brain activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.13.562264v1?rss=1</link>
<description><![CDATA[
Spectral analysis of electroencephalographic (EEG) data simplifies the characterization of periodic band parameters but can obscure underlying dynamics. By contrast, reconstruction of neural activity in state-space preserves geometric complexity in the form of a multidimensional, global attractor. Here we combine these perspectives, inferring complexity and shared dynamics from eigen-time-delay embedding of periodic and aperiodic spectral parameters to yield unique dynamical attractors for each EEG parameter. We find that resting-state alpha and aperiodic attractors show low geometric complexity and shared dynamics with all other frequency bands, what we refer to as geometric cross-parameter coupling. Further, the geometric signatures of alpha and aperiodic attractors dominate spectral dynamics, identifying a geometric core of brain activity. Non-core attractors demonstrate higher complexity but retain traces of this low-dimensional signal, supporting a hypothesis that frequency specific information differentiates out of an integrative, dynamic core. Older adults show lower geometric complexity but greater geometric coupling, resulting from dedifferentiation of gamma band activity. The form and content of resting-state thoughts were further associated with the complexity of core dynamics. Thus, the hallmarks of resting-state EEG in the frequency domain, the alpha peak and the aperiodic backbone, reflect a dynamic, geometric core of resting-state brain activity. This evidence for a geometric core in EEG complements evidence for a regionally defined dynamic core from fMRI-based neuroimaging, further supporting the utility of geometric approaches to the analysis of neural data.
]]></description>
<dc:creator>Pourdavood, P.</dc:creator>
<dc:creator>Jacob, M.</dc:creator>
<dc:date>2023-10-13</dc:date>
<dc:identifier>doi:10.1101/2023.10.13.562264</dc:identifier>
<dc:title><![CDATA[EEG spectral attractors identify a geometric core of resting brain activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.16.562560v1?rss=1">
<title>
<![CDATA[
Cell type specificity of mosaic chromosome 1q gain resolved by snRNA-seq in a case of epilepsy with hyaline protoplasmic astrocytopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.16.562560v1?rss=1</link>
<description><![CDATA[
STRUCTURED ABSTRACTO_ST_ABSIntroductionC_ST_ABSMosaic gain of chromosome 1q (chr1q) has been associated with malformation of cortical development (MCD) and epilepsy. Hyaline protoplasmic astrocytopathy (HPA) is a rare neuropathological finding seen in cases of epilepsy with MCD. The cell-type specificity of mosaic chr1q gain in the brain and the molecular signatures of HPA are unknown.

MethodsWe present a child with pharmacoresistant epilepsy who underwent epileptic focus resections at age 3 and 5 years and was found to have mosaic chr1q gain and HPA. We performed single-nuclei RNA-sequencing (snRNA-seq) of brain tissue from the second resection.

ResultssnRNA-seq showed increased expression of chr1q genes specifically in subsets of neurons and astrocytes. Differentially expressed genes associated with inferred chr1q gain included AKT3 and genes associated with cell adhesion or migration. A subpopulation of astrocytes demonstrated marked enrichment for synapse-associated transcripts, possibly linked to the astrocytic inclusions observed in HPA.

DiscussionsnRNA-seq may be used to infer the cell type-specificity of mosaic chromosomal copy number changes and identify associated gene expression alterations, which in the case of chr1q gain may involve aberrations in cell migration. Future studies using spatial profiling could yield further insights on the molecular signatures of HPA.
]]></description>
<dc:creator>Leng, K.</dc:creator>
<dc:creator>Titus, E.</dc:creator>
<dc:creator>Singhal, N.</dc:creator>
<dc:creator>Cadwell, C. R.</dc:creator>
<dc:creator>Devine, W. P.</dc:creator>
<dc:creator>Tihan, T.</dc:creator>
<dc:creator>Qi, Z.</dc:creator>
<dc:creator>Glenn, O.</dc:creator>
<dc:creator>Kamiya, S.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:creator>Shieh, J.</dc:creator>
<dc:creator>Paredes, M.</dc:creator>
<dc:creator>Upadhyay, V.</dc:creator>
<dc:date>2023-10-17</dc:date>
<dc:identifier>doi:10.1101/2023.10.16.562560</dc:identifier>
<dc:title><![CDATA[Cell type specificity of mosaic chromosome 1q gain resolved by snRNA-seq in a case of epilepsy with hyaline protoplasmic astrocytopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.16.562597v1?rss=1">
<title>
<![CDATA[
The breath shape controls intonation of mouse vocalizations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.16.562597v1?rss=1</link>
<description><![CDATA[
Intonation in speech is the control of vocal pitch to layer expressive meaning to communication, like increasing pitch to indicate a question. Also, stereotyped patterns of pitch are used to create distinct sounds with different denotations, like in tonal languages and, perhaps, the ten sounds in the murine lexicon. A basic tone is created by exhalation through a constricted laryngeal voice box, and it is thought that more complex utterances are produced solely by dynamic changes in laryngeal tension. But perhaps, the shifting pitch also results from altering the swiftness of exhalation. Consistent with the latter model, we describe that intonation in most vocalization types follows deviations in exhalation that appear to be generated by the re-activation of the cardinal breathing muscle for inspiration. We also show that the brainstem vocalization central pattern generator, the iRO, can create this breath pattern. Consequently, ectopic activation of the iRO not only induces phonation, but also the pitch patterns that compose most of the vocalizations in the murine lexicon. These results reveal a novel brainstem mechanism for intonation.
]]></description>
<dc:creator>MacDonald, A.</dc:creator>
<dc:creator>Yackle, K.</dc:creator>
<dc:date>2023-10-17</dc:date>
<dc:identifier>doi:10.1101/2023.10.16.562597</dc:identifier>
<dc:title><![CDATA[The breath shape controls intonation of mouse vocalizations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.17.562797v1?rss=1">
<title>
<![CDATA[
Modulating the Unfolded Protein Response with ISRIB Mitigates Cisplatin Ototoxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.17.562797v1?rss=1</link>
<description><![CDATA[
Cisplatin is a commonly used chemotherapy agent with a nearly universal side effect of sensorineural hearing loss. The cellular mechanisms underlying cisplatin ototoxicity are poorly understood. Efforts in drug development to prevent or reverse cisplatin ototoxicity have largely focused on pathways of oxidative stress and apoptosis. An effective treatment for cisplatin ototoxicity, sodium thiosulfate (STS), while beneficial when used in standard risk hepatoblastoma, is associated with reduced survival in disseminated pediatric malignancies, highlighting the need for more specific drugs without potential tumor protective effects. The unfolded protein response (UPR) and endoplasmic reticulum (ER) stress pathways have been shown to be involved in the pathogenesis of noise-induced hearing loss and cochlear synaptopathy in vivo, and these pathways have been implicated broadly in cisplatin cytotoxicity. This study sought to determine whether the UPR can be targeted to prevent cisplatin ototoxicity. Neonatal cochlear cultures and HEK cells were exposed to cisplatin and UPR-modulating drugs, and UPR marker gene expression and cell death measured. Treatment with ISRIB, a drug that activates eif2B and downregulates the pro-apoptotic PERK/CHOP pathway of the UPR, was tested in an in vivo mouse model of cisplatin ototoxicity and well as a head and neck squamous cell carcinoma (HNSCC) cell-based assay of cisplatin cytotoxicity. Cisplatin exhibited a biphasic, non-linear dose-response of cell death and apoptosis that correlated with different patterns of UPR marker gene expression in HEK cells and cochlear cultures. ISRIB treatment protected against cisplatin-induced hearing loss and hair-cell death, but did not impact cisplatins cytotoxic effects on HNSCC cell viability, unlike STS. These findings demonstrate that targeting the pro-apoptotic PERK/CHOP pathway with ISRIB can mitigate cisplatin ototoxicity without reducing anti-cancer cell effects, suggesting that this may be a viable strategy for drug development.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Rouse, S. L.</dc:creator>
<dc:creator>Matthews, I. R.</dc:creator>
<dc:creator>Sherr, E. H.</dc:creator>
<dc:creator>Chan, D. K.</dc:creator>
<dc:date>2023-10-17</dc:date>
<dc:identifier>doi:10.1101/2023.10.17.562797</dc:identifier>
<dc:title><![CDATA[Modulating the Unfolded Protein Response with ISRIB Mitigates Cisplatin Ototoxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.13.562180v1?rss=1">
<title>
<![CDATA[
Mutant mice lacking alternatively spliced p53 isoforms unveil Ackr4 as a male-specific prognostic factor in Myc-driven B-cell lymphomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.13.562180v1?rss=1</link>
<description><![CDATA[
The Trp53 gene encodes several isoforms of elusive biological significance. Here we show that mice lacking the Trp53 Alternatively Spliced (AS) exon, thereby expressing the canonical p53 protein but not isoforms with the AS C-terminus, have unexpectedly lost a male-specific protection against Myc-induced B-cell lymphomas. Lymphomagenesis was delayed in p53+/+ E-Myc males compared to p53{Delta}AS/{Delta}AS E-Myc males, but also compared to p53+/+ E-Myc and p53{Delta}AS/{Delta}AS E-Myc females. Pre-tumoral splenic cells from p53+/+ E-Myc males exhibited a higher expression of Ackr4, encoding an atypical chemokine receptor with tumor suppressive effects. We identified Ackr4 as a p53 target gene whose p53-mediated transactivation is inhibited by estrogens, and as a male-specific factor of good prognosis relevant for murine E-Myc-induced and human Burkitt lymphomas. Furthermore, the knockout of ACKR4 increased the chemokine-guided migration of Burkitt lymphoma cells. These data demonstrate the functional relevance of alternatively spliced p53 isoforms and reveal sex disparities in Myc-driven lymphomagenesis.
]]></description>
<dc:creator>Fajac, A.</dc:creator>
<dc:creator>Simeonova, I.</dc:creator>
<dc:creator>Leemput, J.</dc:creator>
<dc:creator>Gabriel, M.</dc:creator>
<dc:creator>Morin, A.</dc:creator>
<dc:creator>Lejour, V.</dc:creator>
<dc:creator>Hamon, A.</dc:creator>
<dc:creator>Vaysse-Zinkhofer, W.</dc:creator>
<dc:creator>Eldawra, E.</dc:creator>
<dc:creator>Rakotopare, J.</dc:creator>
<dc:creator>Pinskaya, M.</dc:creator>
<dc:creator>Morillon, A.</dc:creator>
<dc:creator>Bourdon, J.-C.</dc:creator>
<dc:creator>Bardot, B.</dc:creator>
<dc:creator>Toledo, F.</dc:creator>
<dc:date>2023-10-18</dc:date>
<dc:identifier>doi:10.1101/2023.10.13.562180</dc:identifier>
<dc:title><![CDATA[Mutant mice lacking alternatively spliced p53 isoforms unveil Ackr4 as a male-specific prognostic factor in Myc-driven B-cell lymphomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.14.562276v1?rss=1">
<title>
<![CDATA[
Systems-level analyses dissociate genetic regulators of reactive oxygen species and energy production 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.14.562276v1?rss=1</link>
<description><![CDATA[
Respiratory chain dysfunction can decrease ATP and increase reactive oxygen species (ROS) levels. Despite the importance of these metabolic parameters to a wide range of cellular functions and disease, we lack an integrated understanding of how they are differentially regulated. To address this question, we adapted a CRISPRi- and FACS-based platform to compare the effects of respiratory gene knockdown on ROS to their effects on ATP. Focusing on genes whose knockdown is known to decrease mitochondria-derived ATP, we showed that knockdown of genes in specific respiratory chain complexes (I, III and CoQ10 biosynthesis) increased ROS, whereas knockdown of other low ATP hits either had no impact (mitochondrial ribosomal proteins) or actually decreased ROS (complex IV). Moreover, although shifting metabolic conditions profoundly altered mitochondria-derived ATP levels, it had little impact on mitochondrial or cytosolic ROS. In addition, knockdown of a subset of complex I subunits--including NDUFA8, NDUFB4, and NDUFS8--decreased complex I activity, mitochondria-derived ATP and supercomplex level, but knockdown of these genes had differential effects on ROS. Conversely, we found an essential role for ether lipids in the dynamic regulation of mitochondrial ROS levels independent of ATP. Thus, our results identify specific metabolic regulators of cellular ATP and ROS balance that may help dissect the roles of these processes in disease and identify therapeutic strategies to independently target energy failure and oxidative stress.

SignificanceMitochondrial respiration generates both energy (ATP) and reactive oxygen species (ROS). Insufficient energy and increased ROS from respiratory chain dysfunction may be central to the pathophysiology of neurodegenerative diseases and aging. We established a screening platform using CRISPR and fluorescent-cell sorting to compare the impact of decreasing respiratory chain proteins on ROS and ATP levels. The results provide the first systems-level analysis of how ROS and ATP are differentially regulated, and identify genes and respiratory chain complexes that can manipulate each independently. These findings advance our understanding of the relative contributions of ATP and ROS to disease pathophysiology, and guide the development of therapies to preserve energy while minimizing ROS.
]]></description>
<dc:creator>Bennett, N. K.</dc:creator>
<dc:creator>Lee, M.</dc:creator>
<dc:creator>Orr, A. L.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:date>2023-10-18</dc:date>
<dc:identifier>doi:10.1101/2023.10.14.562276</dc:identifier>
<dc:title><![CDATA[Systems-level analyses dissociate genetic regulators of reactive oxygen species and energy production]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.15.561774v1?rss=1">
<title>
<![CDATA[
Development of covalent chemogenetic K2P channel activators 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.15.561774v1?rss=1</link>
<description><![CDATA[
K2P potassium channels regulate excitability by affecting cellular resting membrane potential in the brain, cardiovascular system, immune cells, and sensory organs. Despite their important roles in anesthesia, arrhythmia, pain, hypertension, sleep, and migraine, the ability to control K2P function remains limited. Here, we describe a chemogenetic strategy termed CATKLAMP (Covalent Activation of TREK family K+ channels to cLAmp Membrane Potential) that leverages the discovery of a site in the K2P modulator pocket that reacts with electrophile-bearing derivatives of a TREK subfamily small molecule activator, ML335, to activate the channel irreversibly. We show that the CATKLAMP strategy can be used to probe fundamental aspects of K2P function, as a switch to silence neuronal firing, and is applicable to all TREK subfamily members. Together, our findings exemplify a new means to alter K2P channel activity that should facilitate studies both molecular and systems level studies of K2P function and enable the search for new K2P modulators.
]]></description>
<dc:creator>Deal, P. E.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Mondal, A.</dc:creator>
<dc:creator>Lolicato, M.</dc:creator>
<dc:creator>Furtado de Mandonca, P. R.</dc:creator>
<dc:creator>Black, H.</dc:creator>
<dc:creator>El-Hilali, X.</dc:creator>
<dc:creator>Bryant, C.</dc:creator>
<dc:creator>Isacoff, E. Y.</dc:creator>
<dc:creator>Renslo, A.</dc:creator>
<dc:creator>Minor, D. L.</dc:creator>
<dc:date>2023-10-18</dc:date>
<dc:identifier>doi:10.1101/2023.10.15.561774</dc:identifier>
<dc:title><![CDATA[Development of covalent chemogenetic K2P channel activators]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.16.562637v1?rss=1">
<title>
<![CDATA[
Chromosome size-dependent polar ejection force impairs mammalian mitotic error correction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.16.562637v1?rss=1</link>
<description><![CDATA[
Accurate chromosome segregation requires sister kinetochores to biorient, attaching to opposite spindle poles. To this end, the mammalian kinetochore destabilizes incorrect attachments and stabilizes correct ones, but how it discriminates between these is not yet clear. Here, we test the model that kinetochore tension is the stabilizing cue and ask how chromosome size impacts that model. We live image PtK2 cells, with just 14 chromosomes, widely ranging in size, and find that long chromosomes align at the metaphase plate later than short chromosomes. Enriching for errors and imaging error correction live, we show that long chromosomes exhibit a specific delay in correcting attachments. Using chromokinesin overexpression and laser ablation to perturb polar ejection forces, we find that chromosome size and force on arms determine alignment order. Thus, we propose a model where increased force on long chromosomes can falsely stabilize incorrect attachments, delaying their biorientation. As such, long chromosomes may require compensatory mechanisms for correcting errors to avoid chromosomal instability.
]]></description>
<dc:creator>Chong, M. K.</dc:creator>
<dc:creator>Rosas Salvans, M. K.</dc:creator>
<dc:creator>Tran, V.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2023-10-18</dc:date>
<dc:identifier>doi:10.1101/2023.10.16.562637</dc:identifier>
<dc:title><![CDATA[Chromosome size-dependent polar ejection force impairs mammalian mitotic error correction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.16.562247v1?rss=1">
<title>
<![CDATA[
Multicolor on-chip fluorescence microscopy for real-time surgical guidance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.16.562247v1?rss=1</link>
<description><![CDATA[
In curative-intent cancer surgery, intraoperative fluorescence imaging of both diseased and healthy tissue can help to ensure successful removal of all gross and microscopic disease with minimal damage to neighboring critical structures, such as nerves. Current fluorescence-guided surgery (FGS) systems, however, rely on bulky and rigid optics that incur performance-limiting trade-offs between sensitivity and maneuverability. Moreover, many FGS systems are incapable of multiplexed imaging. As a result, clinical FGS is currently limited to millimeter-scale detection of a single fluorescent target. Here we present a scalable, lens-less fluorescence imaging chip, VISION, capable of sensitive and multiplexed detection within a compact form factor. Central to VISION is a novel optical frontend design combining a low-numerical-aperture fiber optic plate (LNA-FOP) and a multi-bandpass interference filter, which is affixed to a custom CMOS image sensor. The LNA-FOP acts as a planar collimator to improve resolution and compensate for the angle-sensitivity of the interference filter, enabling high-resolution and multiplexed fluorescence imaging without lenses. We show VISION is capable of detecting tumor foci of less than 100 cells at near video framerates and, as proof of principle, can simultaneously visualize both tumor and nerves in ex vivo prostate tissue.
]]></description>
<dc:creator>Roschelle, M. L.</dc:creator>
<dc:creator>Rabbani, R.</dc:creator>
<dc:creator>Papageorgiou, E.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Cooperberg, M.</dc:creator>
<dc:creator>Stohr, B. A.</dc:creator>
<dc:creator>Niknejad, A.</dc:creator>
<dc:creator>Anwar, M.</dc:creator>
<dc:date>2023-10-19</dc:date>
<dc:identifier>doi:10.1101/2023.10.16.562247</dc:identifier>
<dc:title><![CDATA[Multicolor on-chip fluorescence microscopy for real-time surgical guidance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.17.562771v1?rss=1">
<title>
<![CDATA[
Encoding of melody in the human auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.17.562771v1?rss=1</link>
<description><![CDATA[
Melody is a core component of music in which discrete pitches are serially arranged to convey emotion and meaning. Perception of melody varies along several pitch-based dimensions: (1) the absolute pitch of notes, (2) the difference in pitch between successive notes, and (3) the higher-order statistical expectation of each note conditioned on its prior context. While humans readily perceive melody, how these dimensions are collectively represented in the brain and whether their encoding is specialized for music remains unknown. Here, we recorded high-density neurophysiological activity directly from the surface of human auditory cortex while Western participants listened to Western musical phrases. Pitch, pitch-change, and expectation were selectively encoded at different cortical sites, indicating a spatial code for representing distinct dimensions of melody. The same participants listened to spoken English, and we compared evoked responses to music and speech. Cortical sites selective for music were systematically driven by the encoding of expectation. In contrast, sites that encoded pitch and pitch-change used the same neural code to represent equivalent properties of speech. These findings reveal the multidimensional nature of melody encoding, consisting of both music-specific and domain-general sound representations in auditory cortex.

TeaserThe human brain contains both general-purpose and music-specific neural populations for processing distinct attributes of melody.
]]></description>
<dc:creator>Sankaran, N.</dc:creator>
<dc:creator>Leonard, M. K.</dc:creator>
<dc:creator>Theunissen, F. E.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:date>2023-10-19</dc:date>
<dc:identifier>doi:10.1101/2023.10.17.562771</dc:identifier>
<dc:title><![CDATA[Encoding of melody in the human auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.18.562802v1?rss=1">
<title>
<![CDATA[
The genetic evolution of acral melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.18.562802v1?rss=1</link>
<description><![CDATA[
Acral melanoma is an aggressive type of melanoma with unknown origins, arising on the sole, palm, or nail apparatus. It is the most common type of melanoma in individuals with dark skin and is notoriously challenging to treat. Our study examined exome sequencing data from 139 tissue samples, spanning different progression stages, collected from 37 patients. We found that 78.4% of the melanomas displayed one or more clustered copy number transitions with focal amplifications, recurring predominantly on chromosomes 5, 11, 12, and 22. These genomic "hailstorms" were typically shared across all progression stages within individual patients. Genetic alterations known to activate TERT also arose early. By contrast, mutations in the MAP- kinase pathway appeared later during progression, often leading to different tumor areas harboring non-overlapping driver mutations. We conclude that the evolutionary trajectories of acral melanomas substantially diverge from those of melanomas on sun-exposed skin, where MAP-kinase pathway activation initiates the neoplastic cascade followed by immortalization later. The punctuated formation of hailstorms, paired with early TERT activation, suggests a unique mutational mechanism underlying the origins of acral melanoma. Our findings highlight an essential role for telomerase, likely in re-stabilizing tumor genomes after hailstorms have initiated the tumors. The marked genetic heterogeneity, in particular of MAP-kinase pathway drivers, may partly explain the limited success of targeted and other therapies in treating this melanoma subtype.
]]></description>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Fukushima, S.</dc:creator>
<dc:creator>Sheen, Y.-S.</dc:creator>
<dc:creator>Ramelyte, E.</dc:creator>
<dc:creator>Cruz Pacheco, N.</dc:creator>
<dc:creator>Shi, C.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Banik, I.</dc:creator>
<dc:creator>Aquino, J. D.</dc:creator>
<dc:creator>Sangueza Acosta, M.</dc:creator>
<dc:creator>Levesque, M.</dc:creator>
<dc:creator>Dummer, R.</dc:creator>
<dc:creator>Liau, J.-Y.</dc:creator>
<dc:creator>Chu, C.-Y.</dc:creator>
<dc:creator>Shain, A. H.</dc:creator>
<dc:creator>Yeh, I.</dc:creator>
<dc:creator>Bastian, B. C.</dc:creator>
<dc:date>2023-10-19</dc:date>
<dc:identifier>doi:10.1101/2023.10.18.562802</dc:identifier>
<dc:title><![CDATA[The genetic evolution of acral melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.18.562830v1?rss=1">
<title>
<![CDATA[
Modeling the consequences of age-linked rDNA hypermethylation with dCas9-directed DNA methylation in human cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.18.562830v1?rss=1</link>
<description><![CDATA[
Ribosomal DNA (rDNA) genes encode the structural RNAs of the ribosome and are present in hundreds of copies in mammalian genomes. Age-linked DNA hypermethylation throughout the rDNA constitutes a robust "methylation clock" that accurately reports age, yet the consequences of hypermethylation on rDNA function are unknown. We confirmed that pervasive hypermethylation of rDNA occurs during mammalian aging and senescence while rDNA copy number remains stable. We found that DNA methylation is exclusively found on the promoters and gene bodies of inactive rDNA. To model the effects of age-linked methylation on rDNA function, we directed de novo DNA methylation to the rDNA promoter or gene body with a nuclease-dead Cas9 (dCas9) - DNA methyltransferase fusion enzyme in human cells. Hypermethylation at each target site had no detectable effect on rRNA transcription, nucleolar morphology, or cellular growth rate. Instead, human UBF and Pol I remain bound to rDNA promoters in the presence of increased DNA methylation. These data suggest that promoter methylation is not sufficient to impair transcription of the human rDNA and imply that the human rDNA transcription machinery may be resilient to age-linked rDNA hypermethylation.
]]></description>
<dc:creator>Blokhina, Y. P.</dc:creator>
<dc:creator>Buchwalter, A.</dc:creator>
<dc:date>2023-10-19</dc:date>
<dc:identifier>doi:10.1101/2023.10.18.562830</dc:identifier>
<dc:title><![CDATA[Modeling the consequences of age-linked rDNA hypermethylation with dCas9-directed DNA methylation in human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.19.563178v1?rss=1">
<title>
<![CDATA[
Tumor-wide RNA splicing aberrations generate immunogenic public neoantigens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.19.563178v1?rss=1</link>
<description><![CDATA[
T-cell-mediated immunotherapies are limited by the extent to which cancer-specific antigens are homogenously expressed throughout a tumor. We reasoned that recurrent splicing aberrations in cancer represent a potential source of tumor-wide and public neoantigens, and to test this possibility, we developed a novel pipeline for identifying neojunctions expressed uniformly within a tumor across diverse cancer types. Our analyses revealed multiple neojunctions that recur across patients and either exhibited intratumor heterogeneity or, in some cases, were tumor-wide. We identified CD8+ T-cell clones specific for neoantigens derived from tumor-wide and conserved neojunctions in GNAS and RPL22, respectively. TCR-engineered CD8+ T-cells targeting these mutations conferred neoantigen-specific tumor cell eradication. Furthermore, we revealed that cancer-specific dysregulation in splicing factor expression leads to recurrent neojunction expression. Together, these data reveal that a subset of neojunctions are both intratumorally conserved and public, providing the molecular basis for novel T-cell-based immunotherapies that address intratumoral heterogeneity.
]]></description>
<dc:creator>Kwok, D. W.</dc:creator>
<dc:creator>Stevers, N. O.</dc:creator>
<dc:creator>Nejo, T.</dc:creator>
<dc:creator>Chen, L. H.</dc:creator>
<dc:creator>Etxeberria, I.</dc:creator>
<dc:creator>Jung, J.</dc:creator>
<dc:creator>Okada, K.</dc:creator>
<dc:creator>Colton Cove, M.</dc:creator>
<dc:creator>Lakshmanachetty, S.</dc:creator>
<dc:creator>Gallus, M.</dc:creator>
<dc:creator>Barpanda, A.</dc:creator>
<dc:creator>Hong, C.</dc:creator>
<dc:creator>Chan, G. K. L.</dc:creator>
<dc:creator>Wu, S. H.</dc:creator>
<dc:creator>Ramos, E.</dc:creator>
<dc:creator>Yamamichi, A.</dc:creator>
<dc:creator>Liu, J. Y. Z.</dc:creator>
<dc:creator>Watchmaker, P.</dc:creator>
<dc:creator>Ogino, H.</dc:creator>
<dc:creator>Saijo, A.</dc:creator>
<dc:creator>Du, A.</dc:creator>
<dc:creator>Grishanina, N.</dc:creator>
<dc:creator>Woo, J.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:creator>Chang, S. M.</dc:creator>
<dc:creator>Phillips, J. J.</dc:creator>
<dc:creator>Wiita, A.</dc:creator>
<dc:creator>Klebanoff, C. A.</dc:creator>
<dc:creator>Costello, J. F.</dc:creator>
<dc:creator>Okada, H.</dc:creator>
<dc:date>2023-10-20</dc:date>
<dc:identifier>doi:10.1101/2023.10.19.563178</dc:identifier>
<dc:title><![CDATA[Tumor-wide RNA splicing aberrations generate immunogenic public neoantigens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.22.562561v1?rss=1">
<title>
<![CDATA[
Anti-CRISPR proteins trigger a burst of CRISPR-Cas9 expression that enhances phage defense 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.22.562561v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas immune systems provide bacteria with adaptive immunity against bacteriophages, but they are often transcriptionally downregulated to mitigate autoimmunity. In some cases, CRISPR-Cas expression increases in response to a phage infection, but the mechanisms of induction are largely unknown, and it is unclear whether induction occurs strongly and quickly enough to benefit the bacterial host. In S. pyogenes, Cas9 is both an immune effector and autorepressor of CRISPR-Cas expression. Here, we show that phage-encoded anti-CRISPR proteins relieve Cas9 autorepression and trigger a rapid increase in CRISPR-Cas levels during a single phage infective cycle. As a result, fewer cells succumb to lysis leading to a striking survival benefit after multiple rounds of infection. CRISPR-Cas induction also reduces lysogeny, thereby limiting a route for horizontal gene transfer. Altogether, we show that Cas9 is not only a CRISPR-Cas effector and repressor, but also a phage sensor that can mount an anti-anti- CRISPR transcriptional response.
]]></description>
<dc:creator>Workman, R. E.</dc:creator>
<dc:creator>Stoltzfus, M. J.</dc:creator>
<dc:creator>Keith, N. C.</dc:creator>
<dc:creator>Euler, C. W.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Modell, J. W.</dc:creator>
<dc:date>2023-10-22</dc:date>
<dc:identifier>doi:10.1101/2023.10.22.562561</dc:identifier>
<dc:title><![CDATA[Anti-CRISPR proteins trigger a burst of CRISPR-Cas9 expression that enhances phage defense]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.19.563015v1?rss=1">
<title>
<![CDATA[
Context-invariant socioemotional encoding by prefrontal ensembles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.19.563015v1?rss=1</link>
<description><![CDATA[
The prefrontal cortex plays a key role in social interactions, anxiety-related avoidance, and flexible context- dependent behaviors, raising the question: how do prefrontal neurons represent socioemotional information across different environments? Are contextual and socioemotional representations segregated or intermixed, and does this cause socioemotional encoding to remap or generalize across environments? To address this, we imaged neuronal activity in the medial prefrontal cortex of mice engaged in social interactions or anxiety-related avoidance within different environments. Neuronal ensembles representing context and social interaction overlapped more than expected while remaining orthogonal. Anxiety-related representations similarly generalized across environments while remaining orthogonal to contextual information. This shows how prefrontal cortex multiplexes parallel information streams using the same neurons, rather than distinct subcircuits, achieving context-invariant encoding despite context-specific reorganization of population-level activity.
]]></description>
<dc:creator>Frost, N. A.</dc:creator>
<dc:creator>Donohue, K. C.</dc:creator>
<dc:creator>Sohal, V.</dc:creator>
<dc:date>2023-10-23</dc:date>
<dc:identifier>doi:10.1101/2023.10.19.563015</dc:identifier>
<dc:title><![CDATA[Context-invariant socioemotional encoding by prefrontal ensembles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.20.563226v1?rss=1">
<title>
<![CDATA[
Multiomics characterization of cell type repertoires for urine liquid biopsies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.20.563226v1?rss=1</link>
<description><![CDATA[
Urine is assayed alongside blood in medicine, yet current clinical diagnostic tests utilize only a small fraction of its total biomolecular repertoire, potentially foregoing high-resolution insights into human health and disease. In this work, we characterized the joint landscapes of transcriptomic and metabolomic signals in human urine. We also compared the urine transcriptome to plasma cell-free RNA, identifying a distinct cell type repertoire and enrichment for metabolic signal. Untargeted metabolomic measurements identified a complementary set of pathways to the transcriptomic analysis. Our findings suggest that urine is a promising biofluid yielding prognostic and detailed insights for hard-to-biopsy tissues with low representation in the blood, offering promise for a new generation of liquid biopsies.
]]></description>
<dc:creator>Vorperian, S. K.</dc:creator>
<dc:creator>DeFelice, B. C.</dc:creator>
<dc:creator>Buonomo, J. A.</dc:creator>
<dc:creator>Chinchinian, H. J.</dc:creator>
<dc:creator>Gray, I. J.</dc:creator>
<dc:creator>Yan, J.</dc:creator>
<dc:creator>Mach, K. E.</dc:creator>
<dc:creator>La, V.</dc:creator>
<dc:creator>Lee, T. J.</dc:creator>
<dc:creator>Liao, J. C.</dc:creator>
<dc:creator>Lafayette, R.</dc:creator>
<dc:creator>Loeb, G. B.</dc:creator>
<dc:creator>Bertozzi, C. R.</dc:creator>
<dc:creator>Quake, S. R.</dc:creator>
<dc:date>2023-10-23</dc:date>
<dc:identifier>doi:10.1101/2023.10.20.563226</dc:identifier>
<dc:title><![CDATA[Multiomics characterization of cell type repertoires for urine liquid biopsies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.22.563504v1?rss=1">
<title>
<![CDATA[
Neurophysiological evidence of sensory prediction errors driving speech sensorimotor adaptation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.22.563504v1?rss=1</link>
<description><![CDATA[
The human sensorimotor system has a remarkable ability to quickly and efficiently learn movements from sensory experience. A prominent example is sensorimotor adaptation, learning that characterizes the sensorimotor systems response to persistent sensory errors by adjusting future movements to compensate for those errors. Despite being essential for maintaining and fine-tuning motor control, mechanisms underlying sensorimotor adaptation remain unclear. A component of sensorimotor adaptation is implicit (i.e., the learner is unaware of the learning process) which has been suggested to result from sensory prediction errors-the discrepancies between predicted sensory consequences of motor commands and actual sensory feedback. However, to date no direct neurophysiological evidence that sensory prediction errors drive adaptation has been demonstrated. Here, we examined prediction errors via magnetoencephalography (MEG) imaging of the auditory cortex (n = 34) during sensorimotor adaptation of speech to altered auditory feedback, an entirely implicit adaptation task. Specifically, we measured how speaking-induced suppression (SIS)--a neural representation of auditory prediction errors--changed over the trials of the adaptation experiment. SIS refers to the suppression of auditory cortical response to speech onset (in particular, the M100 response) to self-produced speech when compared to the response to passive listening to identical playback of that speech. SIS was reduced (reflecting larger prediction errors) during the early learning phase compared to the initial unaltered feedback phase. Furthermore, reduction in SIS positively correlated with behavioral adaptation extents, suggesting that larger prediction errors were associated with more learning. In contrast, such a reduction in SIS was not found in a control experiment in which participants heard unaltered feedback and thus did not adapt. In addition, in some participants who reached a plateau in the late learning phase, SIS increased (reflecting smaller prediction errors), demonstrating that prediction errors were minimal when there was no further adaptation. Together, these findings provide the first neurophysiological evidence for the hypothesis that prediction errors drive human sensorimotor adaptation.
]]></description>
<dc:creator>Kim, K. S.</dc:creator>
<dc:creator>Hinkley, L. B.</dc:creator>
<dc:creator>Dale, C. L.</dc:creator>
<dc:creator>Nagarajan, S. S.</dc:creator>
<dc:creator>Houde, J. F.</dc:creator>
<dc:date>2023-10-23</dc:date>
<dc:identifier>doi:10.1101/2023.10.22.563504</dc:identifier>
<dc:title><![CDATA[Neurophysiological evidence of sensory prediction errors driving speech sensorimotor adaptation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.23.563616v1?rss=1">
<title>
<![CDATA[
Micrococcin cysteine-to-thiazole conversion through transient interactions between a scaffolding protein and two modification enzymes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.23.563616v1?rss=1</link>
<description><![CDATA[
Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a broad group of compounds mediating microbial competition in nature. Azole/azoline heterocycle formation in the peptide backbone is a key step in the biosynthesis of many RiPPs. Heterocycle formation in RiPP precursors is often carried out by a scaffold protein, an ATP-dependent cyclodehydratase, and an FMN-dependent dehydrogenase. It has generally been assumed that the orchestration of these modifications is carried out by a stable complex including the scaffold, cyclodehydratase and dehydrogenase. The antimicrobial RiPP micrococcin begins as a precursor peptide (TclE) with a 35-amino acid N-terminal leader and a 14-amino acid C-terminal core containing six Cys residues that are converted to thiazoles. The putative scaffold protein (TclI) presumably presents the TclE substrate to a cyclodehydratase (TclJ) and a dehydrogenase (TclN) to accomplish the two-step installation of the six thiazoles. In this study, we identify a minimal TclE leader region required for thiazole formation, we demonstrate complex formation between TclI, TclJ and TclN, and further define regions of these proteins required for complex formation. Our results point to a mechanism of thiazole installation in which TclI associates with the two enzymes in a mutually exclusive fashion, such that each enzyme competes for access to the peptide substrate in a dynamic equilibrium, thus ensuring complete modification of each Cys residue in the TclE core.

IMPORTANCEThiopeptides are a family of antimicrobial peptides characterized for having sulfur-containing heterocycles and for being highly post-translationally modified. Numerous thiopeptides have been identified; almost all of which inhibit protein synthesis in gram-positive bacteria. These intrinsic antimicrobial properties make thiopeptides promising candidates for the development of new antibiotics. The thiopeptide micrococcin is synthesized by the ribosome and undergoes several post-translational modifications (PTMs) to acquire its bioactivity. In this study, we identify key interactions within the enzymatic complex that carries out cysteine to thiazole conversion in the biosynthesis of micrococcin.
]]></description>
<dc:creator>Calvopina-Chavez, D. G.</dc:creator>
<dc:creator>Bursey, D. M.</dc:creator>
<dc:creator>Tseng, Y.-J.</dc:creator>
<dc:creator>Patil, L. M.</dc:creator>
<dc:creator>Bewley, K. D.</dc:creator>
<dc:creator>Bennallack, P. R.</dc:creator>
<dc:creator>McPhie, J. M.</dc:creator>
<dc:creator>Wagstaff, K. B.</dc:creator>
<dc:creator>Daley, A.</dc:creator>
<dc:creator>Miller, S. M.</dc:creator>
<dc:creator>Moody, J. D.</dc:creator>
<dc:creator>Price, J. C.</dc:creator>
<dc:creator>Griffitts, J. S.</dc:creator>
<dc:date>2023-10-23</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563616</dc:identifier>
<dc:title><![CDATA[Micrococcin cysteine-to-thiazole conversion through transient interactions between a scaffolding protein and two modification enzymes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.22.563498v1?rss=1">
<title>
<![CDATA[
Exploring the Roles of RNAs in Chromatin Architecture Using Deep Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.22.563498v1?rss=1</link>
<description><![CDATA[
Recent studies have highlighted the impact of both transcription and transcripts on 3D genome organization, particularly its dynamics. Here, we propose a deep learning framework, called AkitaR, that leverages both genome sequences and genome-wide RNA-DNA interactions to investigate the roles of chromatin-associated RNAs (caRNAs) on genome folding in HFFc6 cells. In order to disentangle the cis- and trans-regulatory roles of caRNAs, we compared models with nascent transcripts, trans-located caRNAs, open chromatin data, or DNA sequence alone. Both nascent transcripts and trans-located caRNAs improved the models predictions, especially at cell-type-specific genomic regions. Analyses of feature importance scores revealed the contribution of caRNAs at TAD boundaries, chromatin loops and nuclear sub-structures such as nuclear speckles and nucleoli to the models predictions. Furthermore, we identified non-coding RNAs (ncRNAs) known to regulate chromatin structures, such as MALAT1 and NEAT1, as well as several novel RNAs, RNY5, RPPH1, POLG-DT and THBS1-IT, that might modulate chromatin architecture through trans-interactions in HFFc6. Our modeling also suggests that transcripts from Alus and other repetitive elements may facilitate chromatin interactions through trans R-loop formation. Our findings provide new insights and generate testable hypotheses about the roles of caRNAs in shaping chromatin organization.
]]></description>
<dc:creator>Kuang, S.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2023-10-24</dc:date>
<dc:identifier>doi:10.1101/2023.10.22.563498</dc:identifier>
<dc:title><![CDATA[Exploring the Roles of RNAs in Chromatin Architecture Using Deep Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.23.563652v1?rss=1">
<title>
<![CDATA[
Neural signatures of stress susceptibility and resilience in the amygdala-hippocampal network 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.23.563652v1?rss=1</link>
<description><![CDATA[
The neural dynamics that underlie divergent anhedonic responses to stress remain unclear. Here, we identified neuronal dynamics in an amygdala-hippocampal circuit that distinguish stress resilience and susceptibility. In a reward-choice task, basolateral amygdala (BLA) activity in resilient mice showed enhanced discrimination of upcoming reward choices. In contrast, a rumination-like signature emerged in the BLA of susceptible mice; a linear decoder could classify the intention to switch or stay on a previously chosen reward. Spontaneous activity in the BLA of susceptible mice was higher dimensional than controls, reflecting the exploration of a larger number of distinct neural states. Manipulation of vCA1-BLA inputs rescued dysfunctional neural dynamics and anhedonia in susceptible mice, suggesting that targeting this pathway can enhance BLA circuit function and ameliorate of depression-related behaviors.

One-Sentence SummaryIdentification and rescue of dysfunctional vCA1-BLA population dynamics and behavior in stress-susceptible mice.
]]></description>
<dc:creator>Xia, F.</dc:creator>
<dc:creator>Fascianelli, V.</dc:creator>
<dc:creator>Vishwakarma, N.</dc:creator>
<dc:creator>Ghinger, F. G.</dc:creator>
<dc:creator>Fusi, S.</dc:creator>
<dc:creator>Kheirbek, M.</dc:creator>
<dc:date>2023-10-24</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563652</dc:identifier>
<dc:title><![CDATA[Neural signatures of stress susceptibility and resilience in the amygdala-hippocampal network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.22.563196v1?rss=1">
<title>
<![CDATA[
Pulmonary Ventilation Analysis Using 1H Ultra-Short Echo Time (UTE) Lung MRI: A Reproducibility Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.22.563196v1?rss=1</link>
<description><![CDATA[
PurposeTo evaluate methods for quantification of pulmonary ventilation with ultrashort echo time (UTE) MRI.

MethodsWe performed a reproducibility study, acquiring two free-breathing 1H UTE lung MRIs on the same day for six healthy volunteers. The 1) 3D + t cyclic b-spline and 2) symmetric image normalization (SyN) methods for image registration were applied after respiratory phase-resolved image reconstruction. Ventilation maps were calculated using 1) Jacobian determinant of the deformation fields minus one, termed regional ventilation, and 2) intensity percentage difference between the registered and fixed image, termed specific ventilation. We compared the reproducibility of all four method combinations via statistical analysis.

ResultsSplit violin plots and Bland-Altman plots are shown for whole lungs and lung sections. The cyclic b-spline registration and Jacobian determinant regional ventilation quantification provide total ventilation volumes that match the segmentation tidal volume, smooth and uniform ventilation maps. The cyclic b-spline registration and specific ventilation combination yields the smallest standard deviation in the Bland-Altman plot.

ConclusionCyclic registration performs better than SyN for respiratory phase-resolved 1H UTE MRI ventilation quantification. Regional ventilation correlates better with segmentation lung volume, while specific ventilation is more reproducible.
]]></description>
<dc:creator>Tan, F.</dc:creator>
<dc:creator>Zhu, X.</dc:creator>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Deveshwar, N.</dc:creator>
<dc:creator>Willmering, M. M.</dc:creator>
<dc:creator>Lustig, M.</dc:creator>
<dc:creator>Larson, P. E. Z.</dc:creator>
<dc:date>2023-10-25</dc:date>
<dc:identifier>doi:10.1101/2023.10.22.563196</dc:identifier>
<dc:title><![CDATA[Pulmonary Ventilation Analysis Using 1H Ultra-Short Echo Time (UTE) Lung MRI: A Reproducibility Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.23.563660v1?rss=1">
<title>
<![CDATA[
A Germline Point Mutation in the MYC-FBW7 Phosphodegron Initiates Hematopoietic Malignancies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.23.563660v1?rss=1</link>
<description><![CDATA[
Oncogenic activation of MYC in cancers predominantly involves increased transcription rather than coding region mutations. However, MYC-dependent lymphomas frequently contain point mutations in the MYC phospho-degron, including at threonine-58 (T58), where phosphorylation permits binding by the FBW7 ubiquitin ligase triggering MYC degradation. To understand how T58 phosphorylation functions in normal cell physiology, we introduced an alanine mutation at T58 (T58A) into the endogenous c-Myc locus in the mouse germline. While MYC-T58A mice develop normally, lymphomas and myeloid leukemias emerge in [~]60% of adult homozygous T58A mice. We find that primitive hematopoietic progenitor cells from MYC-T58A mice exhibit aberrant self-renewal normally associated with hematopoietic stem cells (HSCs) and upregulate a subset of Myc target genes important in maintaining stem/progenitor cell balance. Genomic occupancy by MYC-T58A was increased at all promoters, compared to WT MYC, while genes differentially expressed in a T58A-dependent manner were significantly more proximal to MYC-bound enhancers. MYC-T58A lymphocyte progenitors exhibited metabolic alterations and decreased activation of inflammatory and apoptotic pathways. Our data demonstrate that a single point mutation in Myc is sufficient to produce a profound gain of function in multipotential hematopoietic progenitors associated with self-renewal and initiation of lymphomas and leukemias.
]]></description>
<dc:creator>Freie, B.</dc:creator>
<dc:creator>Carroll, P. A.</dc:creator>
<dc:creator>Varnum-Finney, B. J.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Bernstein, I.</dc:creator>
<dc:creator>Eisenman, R. N.</dc:creator>
<dc:date>2023-10-25</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563660</dc:identifier>
<dc:title><![CDATA[A Germline Point Mutation in the MYC-FBW7 Phosphodegron Initiates Hematopoietic Malignancies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.25.564066v1?rss=1">
<title>
<![CDATA[
The essential chaperone DNAJC17 activates HSP70 to coordinate RNA splicing and G2-M progression. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.25.564066v1?rss=1</link>
<description><![CDATA[
Molecular chaperones including the heat-shock protein 70-kilodalton (HSP70) family and the J-domain containing protein (JDP) co-chaperones maintain homeostatic balance in eukaryotic cells through regulation of the proteome. The expansive JDP family helps direct specific HSP70 functions, and yet loss of single JDP-encoding genes is widely tolerated by mammalian cells, suggesting a high degree of redundancy. By contrast, essential JDPs might carry out HSP70-independent functions or fill cell-context dependent, highly specialized roles within the proteostasis network.

Using a genetic screen of JDPs in human cancer cell lines, we found the RNA recognition motif (RRM) containing DNAJC17 to be pan-essential and investigated the contribution of its structural domains to biochemical and cellular function. We found that the RRM exerts an auto-inhibitory effect on the ability of DNAJC17 to allosterically activate ATP hydrolysis by HSP70. The J-domain, but neither the RRM nor a distal C-terminal alpha helix are required to rescue cell viability after loss of endogenous DNAJC17. Knockdown of DNAJC17 leads to relatively few conserved changes in the abundance of individual mRNAs, but instead deranges gene expression through exon skipping, primarily of genes involved in cell cycle progression. Concordant with cell viability experiments, the C-terminal portions of DNAJC17 are dispensable for restoring splicing and G2-M progression.

Overall, our findings identify essential cellular JDPs and suggest that diversification in JDP structure extends the HSP70-JDP system to control divergent processes such as RNA splicing. Future investigations into the structural basis for auto-inhibition of the DNAJC17 J-domain and the molecular regulation of splicing by these components may provide insights on how conserved biochemical mechanisms can be programmed to fill unique, non-redundant cellular roles and broaden the scope of the proteostasis network.
]]></description>
<dc:creator>Allegakoen, D. V.</dc:creator>
<dc:creator>Kwong, K.</dc:creator>
<dc:creator>Morales, J.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:creator>Sabnis, A. J.</dc:creator>
<dc:date>2023-10-26</dc:date>
<dc:identifier>doi:10.1101/2023.10.25.564066</dc:identifier>
<dc:title><![CDATA[The essential chaperone DNAJC17 activates HSP70 to coordinate RNA splicing and G2-M progression.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.26.564206v1?rss=1">
<title>
<![CDATA[
Designer Fat Cells: Adipogenic Differentiation of CRISPR-Cas9 Genome-Engineered Induced Pluripotent Stem Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.26.564206v1?rss=1</link>
<description><![CDATA[
Adipose tissue is an active endocrine organ that can signal bidirectionally to many tissues and organ systems in the body. With obesity, adipose tissue is a source of low-level inflammation that contributes to various co-morbidities and damage to downstream effector tissues. The ability to synthesize genetically engineered adipose tissue could have critical applications in studying adipokine signaling and the use of adipose tissue for novel therapeutic strategies. This study aimed to develop a method for non-viral adipogenic differentiation of genome-edited murine induced pluripotent stem cells (iPSCs) and to test the ability of such cells to engraft in mice in vivo. Designer adipocytes were created from iPSCs, which can be readily genetically engineered using CRISPR-Cas9 to knock out or insert individual genes of interest. As a model system for adipocyte-based drug delivery, an existing iPSC cell line that transcribes interleukin 1 receptor antagonist under the endogenous macrophage chemoattractant protein-1 promoter was tested for adipogenic capabilities under these same differentiation conditions. To understand the role of various adipocyte subtypes and their impact on health and disease, an efficient method was devised for inducing browning and whitening of IPSC-derived adipocytes in culture. Finally, to study the downstream effects of designer adipocytes in vivo, we transplanted the designer adipocytes into fat-free lipodystrophic mice as a model system for studying adipose signaling in different models of disease or repair. This novel translational tissue engineering and regenerative medicine platform provides an innovative approach to studying the role of adipose interorgan communication in various conditions.
]]></description>
<dc:creator>Ely, E. V.</dc:creator>
<dc:creator>Kapinski, A. T.</dc:creator>
<dc:creator>Paradi, S. G.</dc:creator>
<dc:creator>Tang, R.</dc:creator>
<dc:creator>Guilak, F.</dc:creator>
<dc:creator>Collins, K. H.-M.</dc:creator>
<dc:date>2023-10-26</dc:date>
<dc:identifier>doi:10.1101/2023.10.26.564206</dc:identifier>
<dc:title><![CDATA[Designer Fat Cells: Adipogenic Differentiation of CRISPR-Cas9 Genome-Engineered Induced Pluripotent Stem Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.26.564272v1?rss=1">
<title>
<![CDATA[
Sequence-based machine learning reveals 3D genome differences between bonobos and chimpanzees 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.26.564272v1?rss=1</link>
<description><![CDATA[
Phenotypic divergence between closely related species, including bonobos and chimpanzees (genus Pan), is largely driven by variation in gene regulation. The 3D structure of the genome mediates gene expression; however, genome folding differences in Pan are not well understood. Here, we apply machine learning to predict genome-wide 3D genome contact maps from DNA sequence for 56 bonobos and chimpanzees, encompassing all five extant lineages. We use a pairwise approach to estimate 3D divergence between individuals from the resulting contact maps in 4,420 1 Mb genomic windows. While most pairs were similar,[~] 17% were predicted to be substantially divergent in genome folding. The most dissimilar maps were largely driven by single individuals with rare variants that pro-duce unique 3D genome folding in a region. We also identified 89 genomic windows where bonobo and chimpanzee contact maps substantially diverged, including several windows harboring genes associated with traits implicated in Pan phenotypic divergence. We used in silico mutagenesis to identify 51 3D-modifying variants in these bonobo-chimpanzee diver-gent windows, finding that 34 or 66.67% induce genome folding changes via CTCF binding motif disruption. Our results reveal 3D genome variation at the population-level and identify genomic regions where changes in 3D folding may contribute to phenotypic differences in our closest living relatives.
]]></description>
<dc:creator>Brand, C. M.</dc:creator>
<dc:creator>Kuang, S.</dc:creator>
<dc:creator>Gilbertson, E. N.</dc:creator>
<dc:creator>McArthur, E.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Webster, T. H.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:date>2023-10-26</dc:date>
<dc:identifier>doi:10.1101/2023.10.26.564272</dc:identifier>
<dc:title><![CDATA[Sequence-based machine learning reveals 3D genome differences between bonobos and chimpanzees]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.24.562294v1?rss=1">
<title>
<![CDATA[
AlphaMissense is better correlated with functional assays of missense impact than earlier prediction algorithms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.24.562294v1?rss=1</link>
<description><![CDATA[
Missense variants that alter a single amino acid in the encoded protein contribute to many human disorders but pose a substantial challenge in interpretation. Though these variants can be reliably identified through sequencing, distinguishing the clinically significant ones remains difficult, such that "Variants of Unknown Significance" outnumber those classified as "Pathogenic" or "Likely Pathogenic." Numerous in silico approaches have been developed to predict the functional impact of missense variants to inform clinical interpretation, the latest being AlphaMissense, which uses artificial intelligence methods trained on predicted protein structure. To independently assess the performance of AlphaMissense and 38 other predictors of missense severity, we compared predictions to data from multiplexed assays of variant effect (MAVE). MAVE experiments generate almost every possible individual amino acid change in a gene and measure their functional impact using a high-throughput assay. Assessing 17,696 variants across five genes (DDX3X, MSH2, PTEN, KCNQ4, and BRCA1), we find that AlphaMissense is consistently one of the top five algorithms based on correlation with functional impact and is the best-correlated algorithm for two genes. We conclude that AlphaMissense represents the current best-in-class predictor by this metric; however, the improvement over other algorithms is modest. We note that multiple missense predictors, including AlphaMissense, appear to overcall variants as pathogenic despite minimal functional impact and that substantially more high-quality training data, including consistently analyzed patient cohorts and MAVE analyses, are required to improve accuracy.
]]></description>
<dc:creator>Ljungdahl, A.</dc:creator>
<dc:creator>Kohani, S.</dc:creator>
<dc:creator>Page, N. F.</dc:creator>
<dc:creator>Wells, E. S.</dc:creator>
<dc:creator>Wigdor, E. M.</dc:creator>
<dc:creator>Dong, S.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:date>2023-10-27</dc:date>
<dc:identifier>doi:10.1101/2023.10.24.562294</dc:identifier>
<dc:title><![CDATA[AlphaMissense is better correlated with functional assays of missense impact than earlier prediction algorithms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.24.563645v1?rss=1">
<title>
<![CDATA[
ZNF397 Loss Triggers TET2-driven Epigenetic Rewiring, Lineage Plasticity, and AR-targeted Therapy Resistance in AR-dependent Cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.24.563645v1?rss=1</link>
<description><![CDATA[
Cancer cells exhibit phenotypical plasticity and epigenetic reprogramming, which allows them to evade lineage-dependent targeted treatments by adopting lineage plasticity. The underlying mechanisms by which cancer cells exploit the epigenetic regulatory machinery to acquire lineage plasticity and therapy resistance remain poorly understood. We identified Zinc Finger Protein 397 (ZNF397) as a bona fide co-activator of the androgen receptor (AR), essential for the transcriptional program governing AR-driven luminal lineage. ZNF397 deficiency facilitates the transition of cancer cell from an AR-driven luminal lineage to a Ten-Eleven Translocation 2 (TET2)-driven lineage plastic state, ultimately promoting resistance to therapies inhibiting AR signaling. Intriguingly, our findings indicate that TET2 inhibitor can eliminate the AR targeted therapies resistance in ZNF397-deficient tumors. These insights uncover a novel mechanism through which prostate and breast cancers acquire lineage plasticity via epigenetic rewiring and offer promising implications for clinical interventions designed to overcome therapy resistance dictated by lineage plasticity.

Statement of SignificanceThis study reveals a novel epigenetic mechanism regulating tumor lineage plasticity and therapy response, enhances understanding of drug resistance and unveils a new therapeutic strategy for prostate cancer and other malignancies. Our findings also illuminate TET2s oncogenic role and mechanistically connect TET2-driven epigenetic rewiring to lineage plasticity and therapy resistance.
]]></description>
<dc:creator>Mu, P.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Sjostrom, M.</dc:creator>
<dc:creator>Deng, S.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Jonhson, N.</dc:creator>
<dc:creator>Gonzalez, J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Metang, L.</dc:creator>
<dc:creator>Rodriguez, C.</dc:creator>
<dc:creator>Mukherji, A.</dc:creator>
<dc:creator>Wainwright, G.</dc:creator>
<dc:creator>Yu, X.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Barnes, S.</dc:creator>
<dc:creator>Hofstad, M.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Hanker, A.</dc:creator>
<dc:creator>He, H.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Raj, G.</dc:creator>
<dc:creator>Arteaga, C.</dc:creator>
<dc:creator>Feng, F.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:date>2023-10-27</dc:date>
<dc:identifier>doi:10.1101/2023.10.24.563645</dc:identifier>
<dc:title><![CDATA[ZNF397 Loss Triggers TET2-driven Epigenetic Rewiring, Lineage Plasticity, and AR-targeted Therapy Resistance in AR-dependent Cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.26.564156v1?rss=1">
<title>
<![CDATA[
Challenges in the discovery of tumor-specific alternative splicing-derived cell-surface antigens in glioma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.26.564156v1?rss=1</link>
<description><![CDATA[
BackgroundDespite advancements in cancer immunotherapy, solid tumors remain formidable challenges. In glioma, profound inter-and intra-tumoral heterogeneity of antigen landscape hampers therapeutic development. Therefore, it is critical to consider alternative sources to expand the repertoire of targetable (neo-)antigens and improve therapeutic outcomes. Accumulating evidence suggests that tumor-specific alternative splicing (AS) could be an untapped reservoir of neoantigens.

ResultsIn this study, we investigated tumor-specific AS events in glioma, focusing on those predicted to generate major histocompatibility complex (MHC)-presentation-independent, cell-surface neoantigens that could be targeted by antibodies and chimeric antigen receptor (CAR)-T cells. We systematically analyzed bulk RNA-sequencing datasets comparing 429 tumor samples (from The Cancer Genome Atlas [TCGA]) and 9,166 normal tissue samples (from the Genotype-Tissue Expression project [GTEx]), and identified 13 AS events in 7 genes predicted to be expressed in more than 10% of the patients, including PTPRZ1 and BCAN, which were corroborated by an external RNA-sequencing dataset. Subsequently, we validated our predictions and elucidated the complexity of the isoforms using full-length transcript amplicon sequencing on patient-derived glioblastoma cells. However, analyses of the RNA-sequencing datasets of spatially mapped and longitudinally collected clinical tumor samples unveiled remarkable spatiotemporal heterogeneity of the candidate AS events. Furthermore, proteomics analysis did not reveal any peptide spectra matching the putative neoantigens.

ConclusionsOur investigation illustrated the diverse characteristics of the tumor-specific AS events and the challenges of antigen exploration due to their notable spatiotemporal heterogeneity and elusive nature at the protein levels. Redirecting future efforts toward intracellular, MHC-presented antigens could offer a more viable avenue.
]]></description>
<dc:creator>Nejo, T.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Wang, A.</dc:creator>
<dc:creator>Lakshmanachetty, S.</dc:creator>
<dc:creator>Gallus, M.</dc:creator>
<dc:creator>Kwok, D. W.</dc:creator>
<dc:creator>Hong, C.</dc:creator>
<dc:creator>Chen, L. G.</dc:creator>
<dc:creator>Carrera, D. A.</dc:creator>
<dc:creator>Zhang, M. Y.</dc:creator>
<dc:creator>Stevers, N. O.</dc:creator>
<dc:creator>Maldonado, G. C.</dc:creator>
<dc:creator>Yamamichi, A.</dc:creator>
<dc:creator>Watchmaker, P.</dc:creator>
<dc:creator>Naik, A.</dc:creator>
<dc:creator>Shai, A.</dc:creator>
<dc:creator>Phillips, J.</dc:creator>
<dc:creator>Chang, S. M.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:creator>Costello, J. F.</dc:creator>
<dc:creator>Diaz, A. A.</dc:creator>
<dc:creator>Okada, H.</dc:creator>
<dc:date>2023-10-27</dc:date>
<dc:identifier>doi:10.1101/2023.10.26.564156</dc:identifier>
<dc:title><![CDATA[Challenges in the discovery of tumor-specific alternative splicing-derived cell-surface antigens in glioma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.26.564222v1?rss=1">
<title>
<![CDATA[
Altered PBP4 and GdpP functions synergistically mediate MRSA-like high-level, broad-spectrum β-lactam resistance in Staphylococcus aureus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.26.564222v1?rss=1</link>
<description><![CDATA[
Infections caused by Staphylococcus aureus are a leading cause of mortality worldwide. S. aureus infections caused by Methicillin-Resistant Staphylococcus aureus (MRSA) are particularly difficult to treat due to their resistance to Next Generation {beta}-lactams (NGB) such as Methicillin, Nafcillin, Oxacillin etc. Resistance to NGBs, which is alternatively known as broad-spectrum {beta}- lactam resistance is classically mediated by PBP2a, a Penicillin-Binding Protein encoded by mecA (or mecC) in MRSA. Thus, presence of mec genes among S. aureus serves as the predictor of resistance to NGBs and facilitates determination of the proper therapeutic strategy for a staphylococcal infection. Although far less appreciated, mecA deficient S. aureus strains can also exhibit NGB resistance. These strains, which are collectively termed as Methicillin-Resistant Lacking mec (MRLM) are currently being identified in increasing numbers among natural resistant isolates of S. aureus. The mechanism/s through which MRLMs produce resistance to NGBs remains unknown. In this study, we demonstrate that mutations that alter PBP4 and GdpP functions, which are often present among MRLMs can synergistically mediate resistance to NGBs. Furthermore, our results unravel that this novel mechanism potentially enables MRLMs to produce resistance towards NGBs at levels comparable to that of MRSAs. Our study, provides a fresh new perspective about alternative mechanisms of NGBs resistance, challenging our current overall understanding of high-level, broad-spectrum {beta}-lactam resistance in S. aureus. It thus suggests reconsideration of the current approach towards diagnosis and treatment of {beta}-lactam resistant S. aureus infections.
]]></description>
<dc:creator>Lai, L.-Y.</dc:creator>
<dc:creator>Satishkumar, N.</dc:creator>
<dc:creator>Cardozo, S.</dc:creator>
<dc:creator>Hemmadi, V.</dc:creator>
<dc:creator>Marques, L. B.</dc:creator>
<dc:creator>Huang, L.</dc:creator>
<dc:creator>Filipe, S. R.</dc:creator>
<dc:creator>Pinho, M. G.</dc:creator>
<dc:creator>Chambers, H. F.</dc:creator>
<dc:creator>Chatterjee, S. S.</dc:creator>
<dc:date>2023-10-28</dc:date>
<dc:identifier>doi:10.1101/2023.10.26.564222</dc:identifier>
<dc:title><![CDATA[Altered PBP4 and GdpP functions synergistically mediate MRSA-like high-level, broad-spectrum β-lactam resistance in Staphylococcus aureus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.24.562292v1?rss=1">
<title>
<![CDATA[
Rosace: a robust deep mutational scanning analysis framework employing position and mean-variance shrinkage 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.24.562292v1?rss=1</link>
<description><![CDATA[
Deep mutational scanning (DMS) enables functional insight into protein mutations with multiplexed measurements of thousands of genetic variants in a protein simultaneously. The small sample size of DMS renders classical statistical methods ineffective, for example, p-values cannot be correctly calibrated when treating variants independently. We propose Rosace, a Bayesian framework for analyzing growth-based deep mutational scanning data. Rosace leverages amino acid position information to increase power and control the false discovery rate by sharing information across parameters via shrinkage. To benchmark Rosace against existing methods, we developed Rosette, a simulation framework that simulates the distributional properties of DMS. Further, we show that Rosace is robust to the violation of model assumptions and is more powerful than existing tools under Rosette simulation and real data.
]]></description>
<dc:creator>Rao, J.</dc:creator>
<dc:creator>Xin, R.</dc:creator>
<dc:creator>Macdonald, C.</dc:creator>
<dc:creator>Howard, M.</dc:creator>
<dc:creator>Estevam, G. O.</dc:creator>
<dc:creator>Yee, S. W.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Coyote-Maestas, W.</dc:creator>
<dc:creator>Pimentel, H.</dc:creator>
<dc:date>2023-10-29</dc:date>
<dc:identifier>doi:10.1101/2023.10.24.562292</dc:identifier>
<dc:title><![CDATA[Rosace: a robust deep mutational scanning analysis framework employing position and mean-variance shrinkage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.24.563768v1?rss=1">
<title>
<![CDATA[
DREDge: robust motion correction for high-density extracellular recordings across species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.24.563768v1?rss=1</link>
<description><![CDATA[
High-density microelectrode arrays (MEAs) have opened new possibilities for systems neuroscience in human and non-human animals, but brain tissue motion relative to the array poses a challenge for downstream analyses, particularly in human recordings. We introduce DREDge (Decentralized Registration of Electrophysiology Data), a robust algorithm which is well suited for the registration of noisy, nonstationary extracellular electrophysiology recordings. In addition to estimating motion from spikes in the action potential (AP) frequency band, DREDge enables automated tracking of motion at high temporal resolution in the local field potential (LFP) frequency band. In human intraoperative recordings, which often feature fast (period <1s) motion, DREDge correction in the LFP band enabled reliable recovery of evoked potentials, and significantly reduced single-unit spike shape variability and spike sorting error. Applying DREDge to recordings made during deep probe insertions in nonhuman primates demonstrated the possibility of tracking probe motion of centimeters across several brain regions while simultaneously mapping single unit electrophysiological features. DREDge reliably delivered improved motion correction in acute mouse recordings, especially in those made with an recent ultra-high density probe. We also implemented a procedure for applying DREDge to recordings made across tens of days in chronic implantations in mice, reliably yielding stable motion tracking despite changes in neural activity across experimental sessions. Together, these advances enable automated, scalable registration of electrophysiological data across multiple species, probe types, and drift cases, providing a stable foundation for downstream scientific analyses of these rich datasets.
]]></description>
<dc:creator>Windolf, C.</dc:creator>
<dc:creator>Yu, H.</dc:creator>
<dc:creator>Paulk, A. C.</dc:creator>
<dc:creator>Meszena, D.</dc:creator>
<dc:creator>Munoz, W.</dc:creator>
<dc:creator>Boussard, J.</dc:creator>
<dc:creator>Hardstone, R.</dc:creator>
<dc:creator>Caprara, I.</dc:creator>
<dc:creator>Jamali, M.</dc:creator>
<dc:creator>Kfir, Y.</dc:creator>
<dc:creator>Xu, D.</dc:creator>
<dc:creator>Chung, J. E.</dc:creator>
<dc:creator>Sellers, K. K.</dc:creator>
<dc:creator>Ye, Z.</dc:creator>
<dc:creator>Shaker, J.</dc:creator>
<dc:creator>Lebedeva, A.</dc:creator>
<dc:creator>Raghavan, M.</dc:creator>
<dc:creator>Trautmann, E.</dc:creator>
<dc:creator>Melin, M. D.</dc:creator>
<dc:creator>Couto, J.</dc:creator>
<dc:creator>Garcia, S.</dc:creator>
<dc:creator>Coughlin, B.</dc:creator>
<dc:creator>Horvath, C.</dc:creator>
<dc:creator>Fiath, R.</dc:creator>
<dc:creator>Ulbert, I.</dc:creator>
<dc:creator>Movshon, J. A.</dc:creator>
<dc:creator>Shadlen, M. N.</dc:creator>
<dc:creator>Churchland, M. M.</dc:creator>
<dc:creator>Churchland, A. K.</dc:creator>
<dc:creator>Steinmetz, N. A.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Schweitzer, J. S.</dc:creator>
<dc:creator>Williams, Z. M.</dc:creator>
<dc:creator>Cash, S. S.</dc:creator>
<dc:creator>Paninski, L.</dc:creator>
<dc:creator>Varol, E.</dc:creator>
<dc:date>2023-10-29</dc:date>
<dc:identifier>doi:10.1101/2023.10.24.563768</dc:identifier>
<dc:title><![CDATA[DREDge: robust motion correction for high-density extracellular recordings across species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.25.564029v1?rss=1">
<title>
<![CDATA[
Structure-Based Discovery of a NPFF1R Antagonist with Analgesic Activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.25.564029v1?rss=1</link>
<description><![CDATA[
While opioid drugs remain among the most effective analgesics for pain management, adverse effects limit their use. Molecules that synergize with opioids, increasing analgesia without increasing side effects, could prove beneficial. A potential way to do so is via the RF-amide receptor system, as NPFFR1 agonists reduce {micro}- opioid receptor ({micro}OR)-based analgesia while antagonists increase it. These inferences are, however, clouded by the lack of selectivity of most NPFF1R ligands. Seeking selective antagonists of the NPFF1R, we screened a large virtual library against a homology model of NPFF1R. From 26 high-ranking molecules that were synthesized and tested, one antagonized NPFF1R with a Ki of 319 nM. Structure-based optimization led to a 22 nM antagonist of NPFF1R, compound 56, with selectivity against a large panel of GPCRs. When administered alone, 56 has no activity in mouse tail-flick nociception assays. However, coadministration of compound 56 and morphine produced significantly greater antinociception than did morphine alone, consistent with the notion that NPFF1R nociceptive activity occurs via modulation of {micro}OR signaling. Surprisingly, in the hot-plate assays 56 was analgesic by itself, suggesting that NPFF1R alone can also confer analgesia. At equi-analgesic doses, combinations of 56 with morphine reduced the common constipation side effect of morphine versus using morphine alone. The high selectivity of 56 and its activity in cooperation with morphine supports further analgesic development against NPFF1R and against the RF-amide family of receptors more generally.
]]></description>
<dc:creator>Bender, B. J.</dc:creator>
<dc:creator>Pickett, J. E.</dc:creator>
<dc:creator>Braz, J.</dc:creator>
<dc:creator>Kang, H. J.</dc:creator>
<dc:creator>Gahbauer, S.</dc:creator>
<dc:creator>Bhardwaj, K.</dc:creator>
<dc:creator>Rodriguez-Rosado, S.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Jain, M.</dc:creator>
<dc:creator>Basbaum, A.</dc:creator>
<dc:creator>Roth, B. L.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2023-10-29</dc:date>
<dc:identifier>doi:10.1101/2023.10.25.564029</dc:identifier>
<dc:title><![CDATA[Structure-Based Discovery of a NPFF1R Antagonist with Analgesic Activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.27.564400v1?rss=1">
<title>
<![CDATA[
Docking for molecules that bind in a symmetric stack with SymDOCK 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.27.564400v1?rss=1</link>
<description><![CDATA[
Discovering ligands for amyloid fibrils, such as those formed by the tau protein, is an area of much current interest. In recent structures, ligands bind in stacks in the tau fibrils to reflect the rotational and translational symmetry of the fibril itself; in these structures the ligands make few interactions with the protein but interact extensively with each other. To exploit this symmetry and stacking, we developed SymDOCK, a method to dock molecules that follow the proteins symmetry. For each prospective ligand pose, we apply the symmetry operation of the fibril to generate a self-interacting and fibril-interacting stack, checking that doing so will not cause a clash between the original molecule and its image. Absent a clash, we retain that pose and add the ligand-ligand van der Waals energy to the ligands docking score (here using DOCK3.8). We can check these geometries and energies using an implementation of ANI, a neural network-based quantum-mechanical evaluation of the ligand stacking energies. In retrospective calculations, symmetry docking can reproduce the poses of three tau PET tracers whose structures have been determined. More convincingly, in a prospective study SymDOCK predicted the structure of the PET tracer MK-6240 bound in a symmetrical stack to AD PHF tau before that structure was determined; the docked pose was used to determine how MK-6240 fit the cryo-EM density. In proof-of-concept studies, SymDOCK enriched known ligands over property-matched decoys in retrospective screens without sacrificing docking speed, and can address large library screens that seek new symmetrical stackers. Future applications of this approach will be considered.
]]></description>
<dc:creator>Smith, M. S.</dc:creator>
<dc:creator>Knight, I. S.</dc:creator>
<dc:creator>Kormos, R. C.</dc:creator>
<dc:creator>Pepe, J. G.</dc:creator>
<dc:creator>Kunach, P.</dc:creator>
<dc:creator>Diamond, M. I.</dc:creator>
<dc:creator>Shahmoradian, S. H.</dc:creator>
<dc:creator>Irwin, J. I.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2023-10-29</dc:date>
<dc:identifier>doi:10.1101/2023.10.27.564400</dc:identifier>
<dc:title><![CDATA[Docking for molecules that bind in a symmetric stack with SymDOCK]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.28.564055v1?rss=1">
<title>
<![CDATA[
Multi-scale photocatalytic proximity labeling reveals cell surface neighbors on and between cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.28.564055v1?rss=1</link>
<description><![CDATA[
The cell membrane proteome is the primary biohub for cell communication, yet we are only beginning to understand the dynamic protein neighborhoods that form on the cell surface and between cells. Proximity labeling proteomics (PLP) strategies using chemically reactive probes are powerful approaches to yield snapshots of protein neighborhoods but are currently limited to one single resolution based on the probe labeling radius. Here, we describe a multi-scale PLP method with tunable resolution using a commercially available histological dye, Eosin Y, which upon visible light illumination, activates three different photo-probes with labeling radii ranging from [~]100 to 3000 [A]. We applied this platform to profile neighborhoods of the oncogenic epidermal growth factor receptor (EGFR) and orthogonally validated >20 neighbors using immuno-assays and AlphaFold-Multimer prediction that generated plausible binary interaction models. We further profiled the protein neighborhoods of cell-cell synapses induced by bi-specific T-cell engagers (BiTEs) and chimeric antigen receptor (CAR)T cells at longer length scales. This integrated multi-scale PLP platform maps local and distal protein networks on cell surfaces and between cells. We believe this information will aid in the systematic construction of the cell surface interactome and reveal new opportunities for immunotherapeutics.
]]></description>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Schaefer, K.</dc:creator>
<dc:creator>Lui, I.</dc:creator>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Fossati, A. A.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Palar, A.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2023-10-29</dc:date>
<dc:identifier>doi:10.1101/2023.10.28.564055</dc:identifier>
<dc:title><![CDATA[Multi-scale photocatalytic proximity labeling reveals cell surface neighbors on and between cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.26.564203v1?rss=1">
<title>
<![CDATA[
Effects of chronic lithium treatment on neuronal excitability and GABAergic transmission in an Ank3 mutant mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.26.564203v1?rss=1</link>
<description><![CDATA[
Bipolar disorder (BD) is a prevalent psychiatric condition characterized by mood dysregulation, psychosocial impairment, and an increased risk of suicide. The gene ANK3 has been identified as a risk locus for BD through multiple genome-wide association studies (GWAS). However, the mechanisms by which ANK3 variants influence BD pathophysiology and treatment response remain unclear. ANK3 encodes ankyrin-G, a protein that organizes the axon initial segment (AIS) and nodes of Ranvier by scaffolding ion channels and cell adhesion molecules to the cytoskeleton. Recent studies show that ankyrin-G interacts with the GABAA receptor-associated protein (GABARAP) to stabilize inhibitory synapses, potentially linking ANK3 variants to inhibitory (GABAergic) signaling deficits associated with BD. We previously demonstrated that the BD-associated variant, ANK3 p.W1989R, disrupts the ankyrin-G/ GABARAP interaction, resulting in inhibitory deficits and cortical pyramidal neuron hyperexcitability in mice. In this study, we investigate how lithium, a common BD therapeutic, modulates neuronal excitability in this model. Our findings show that chronic lithium treatment selectively enhances presynaptic GABAergic neurotransmission, reduces neuronal hyperexcitability, and partially rescues AIS length, without altering the density of GABAergic synapses. We also show that the selective glycogen synthase kinase-3 beta (GSK-3{beta}) inhibitor Tideglusib recapitulates the enhancement of presynaptic GABAergic signaling. These findings shed new light on how ANK3 variants may contribute to inhibitory deficits in BD and demonstrate that lithium treatment is able to restore these deficits, likely through GSK-3{beta} inhibition. Furthermore, these findings highlight GSK-3{beta} inhibition as a promising therapeutic strategy for treating BD and other neurological disorders affected by GABAergic dysfunction.
]]></description>
<dc:creator>Caballero-Floran, R. N.</dc:creator>
<dc:creator>Nelson, A. D.</dc:creator>
<dc:creator>Min, L.</dc:creator>
<dc:creator>Jenkins, P. M.</dc:creator>
<dc:date>2023-10-30</dc:date>
<dc:identifier>doi:10.1101/2023.10.26.564203</dc:identifier>
<dc:title><![CDATA[Effects of chronic lithium treatment on neuronal excitability and GABAergic transmission in an Ank3 mutant mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.27.564435v1?rss=1">
<title>
<![CDATA[
Colloidal aggregation confounds cell-based Covid-19 antiviral screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.27.564435v1?rss=1</link>
<description><![CDATA[
Colloidal aggregation is one of the largest contributors to false-positives in early drug discovery and chemical biology. Much work has focused on its impact on pure-protein screens; here we consider aggregations role in cell-based infectivity assays in Covid-19 drug repurposing. We began by investigating the potential aggregation of 41 drug candidates reported as SARs-CoV-2 entry inhibitors. Of these, 17 formed colloidal-particles by dynamic light scattering and exhibited detergent-dependent enzyme inhibition. To evaluate antiviral efficacy of the drugs in cells we used spike pseudotyped lentivirus and pre-saturation of the colloids with BSA. The antiviral potency of the aggregators was diminished by at least 10-fold and often entirely eliminated in the presence of BSA, suggesting antiviral activity can be attributed to the non-specific nature of the colloids. In confocal microscopy, the aggregates induced fluorescent puncta of labeled spike protein, consistent with sequestration of the protein on the colloidal particles. Addition of either non-ionic detergent or of BSA disrupted these puncta. These observations suggest that colloidal aggregation is common among cell-based anti-viral drug repurposing, and perhaps cell-based assays more broadly, and offers rapid counter-screens to detect and eliminate these artifacts, allowing the community invest resources in compounds with true potential as a Covid-19 therapeutic.
]]></description>
<dc:creator>Glenn, I. S.</dc:creator>
<dc:creator>Hall, L. N.</dc:creator>
<dc:creator>Khalid, M. M.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2023-10-30</dc:date>
<dc:identifier>doi:10.1101/2023.10.27.564435</dc:identifier>
<dc:title><![CDATA[Colloidal aggregation confounds cell-based Covid-19 antiviral screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.27.564420v1?rss=1">
<title>
<![CDATA[
Immunosuppression is a conserved driver of tuberculosis susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.27.564420v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) causes 1.25 million deaths a year. However, tuberculosis (TB) pathogenesis remains poorly understood and is not fully recapitulated in standard mouse models. Here we find that gene signatures from three different Mtb-susceptible mouse models predict active TB disease in humans significantly better than a signature from resistant C57BL/6 (B6) mice. Conserved among susceptible mice, non-human primates, and humans, but largely absent from B6 mice, was Mtb-induced differentiation of macrophages into an Spp1+ differentiation state. Spp1+ macrophages expressed high levels of immunosuppressive molecules including IL-1 receptor antagonist (IL-1Ra). IL-1Ra was previously reported to cause Mtb susceptibility in one mouse model, but whether IL-1Ra is broadly important remains uncertain. Here we report that enhancement of IL-1 signaling via deletion of IL-Ra promoted bacterial control across three susceptible mouse models. We found IL-1 signaling amplified production of multiple cytokines by lymphoid and stromal cells, providing a multifactorial mechanism for how IL-1 promotes Mtb control. Our results indicate that myeloid cell expression of immunosuppressive molecules, in particular IL-1 receptor antagonist, is a conserved early mechanism limiting Mtb control in mice, non-human primates, and humans.
]]></description>
<dc:creator>Kotov, D. I.</dc:creator>
<dc:creator>Lee, O. V.</dc:creator>
<dc:creator>Ji, D. X.</dc:creator>
<dc:creator>Jaye, D. L.</dc:creator>
<dc:creator>Suliman, S.</dc:creator>
<dc:creator>Gabay, C.</dc:creator>
<dc:creator>Vance, R. E.</dc:creator>
<dc:date>2023-11-01</dc:date>
<dc:identifier>doi:10.1101/2023.10.27.564420</dc:identifier>
<dc:title><![CDATA[Immunosuppression is a conserved driver of tuberculosis susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.30.563998v1?rss=1">
<title>
<![CDATA[
Efficient semi-supervised semantic segmentation of electron microscopy cancer images with sparse annotations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.30.563998v1?rss=1</link>
<description><![CDATA[
Electron microscopy (EM) enables imaging at nanometer resolution and can shed light on how cancer evolves to develop resistance to therapy. Acquiring these images has become a routine task; however, analyzing them is now the bottleneck, as manual structure identification is very time-consuming and can take up to several months for a single sample. Deep learning approaches offer a suitable solution to speed up the analysis. In this work, we present a study of several state-of-the-art deep learning models for the task of segmenting nuclei and nucleoli in volumes from tumor biopsies. We compared previous results obtained with the ResUNet architecture to the more recent UNet++, FracTALResNet, SenFormer, and CEECNet models. In addition, we explored the utilization of unlabeled images through semi-supervised learning with Cross Pseudo Supervision. We have trained and evaluated all of the models on sparse manual labels from three fully annotated in-house datasets that we have made available on demand, demonstrating improvements in terms of 3D Dice score. From the analysis of these results, we drew conclusions on the relative gains of using more complex models, semi-supervised learning as well as next steps for the mitigation of the manual segmentation bottleneck.
]]></description>
<dc:creator>Pagano, L.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Bousselham, W.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:date>2023-11-01</dc:date>
<dc:identifier>doi:10.1101/2023.10.30.563998</dc:identifier>
<dc:title><![CDATA[Efficient semi-supervised semantic segmentation of electron microscopy cancer images with sparse annotations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.31.560822v1?rss=1">
<title>
<![CDATA[
Critical role of CD206+ macrophages in organizing anti-tumor immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.31.560822v1?rss=1</link>
<description><![CDATA[
Tumor-associated macrophages (TAMs) are frequently categorized as being  M1 or  M2 polarized, even as substantial data challenges this binary modeling of macrophage cell state. One molecule consistently referenced as a delineator of a putative immunosuppressive  M2 state is the surface protein CD206. We thus made a novel conditional CD206 (Mrc1) knock-in mouse to specifically visualize and/or deplete CD206+  M2-like TAMs and assess their correspondence with pro-tumoral immunity. Early, but not late depletion of CD206+ macrophages and monocytes (here,  Mono/Macs) led to an indirect loss of a key anti-tumor network of NK cells, conventional type I dendritic cells (cDC1) and CD8 T cells. Among myeloid cells, we found that the CD206+ TAMs are the primary producers of CXCL9, and able to differentially attract activated CD8 T cells. In contrast, a population of stress-responsive TAMs ("Hypoxic" or Spp1+) and immature monocytes, which lack CD206 expression and become prominent following early depletion, expressed markedly diminished levels of CXCL9. Those NK and CD8 T cells which enter CD206- depleted tumors express vastly reduced levels of the corresponding receptor Cxcr3, the cDC1- attracting chemokine Xcl1 and cDC1 growth factor Flt3l transcripts. Consistent with the loss of this critical network, early CD206+ TAM depletion decreased tumor control by antigen specific CD8 T cells in mice. Likewise, in humans, the CD206Replete, but not the CD206Depleted Mono/Mac gene signature correlated robustly with CD8 T cell, NK cell and stimulatory cDC1 gene signatures and transcriptomic signatures skewed towards CD206Replete Mono/Macs associated with better survival. Together, these findings negate the unqualified classification of CD206+  M2-like macrophages as immunosuppressive by illuminating contexts for their role in organizing a critical tumor-reactive archetype of immunity.
]]></description>
<dc:creator>Ray, A.</dc:creator>
<dc:creator>Hu, K. H.</dc:creator>
<dc:creator>Kersten, K.</dc:creator>
<dc:creator>Kuhn, N. F.</dc:creator>
<dc:creator>Samad, B.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2023-11-02</dc:date>
<dc:identifier>doi:10.1101/2023.10.31.560822</dc:identifier>
<dc:title><![CDATA[Critical role of CD206+ macrophages in organizing anti-tumor immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.31.564826v1?rss=1">
<title>
<![CDATA[
Using behavioral biomarkers to redefine epochs of spontaneous recovery following spinal cord injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.31.564826v1?rss=1</link>
<description><![CDATA[
The brain-spinal cord axis generates movement by assembling motor primitives into coordinated sequences. Spinal cord injury (SCI) disrupts this neuroaxis, impairing not only locomotion, but the full repertoire of behavior. Traditional scales for quantifying recovery collapse this complexity into predefined locomotor-focused criteria that obscure heterogeneity in recovery. To quantify the full behavioral repertoire following SCI, we adapted motion sequencing (MoSeq) to identify sub-second behavioral "syllables" and capture their usage and sequential organization without predefined features. We identified biomechanically distinct variants within syllable classes that are shared across injury severities and found that recovery is jointly structured by injury severity and individual mouse identity. Changes in sequences, however, unfold along a conserved temporal trajectory. By compressing behavior into a single metric, we uncovered clusters of coevolving locomotor and non-locomotor behaviors. These results frame SCI recovery with repertoire-level changes, where adaptive strategies emerge from constrained access to motor primitives and their sequences.
]]></description>
<dc:creator>Eisdorfer, J. T.</dc:creator>
<dc:creator>Thackray, J.</dc:creator>
<dc:creator>Theis, T.</dc:creator>
<dc:creator>Vivinetto, A.</dc:creator>
<dc:creator>Ricci, M. T.</dc:creator>
<dc:creator>Lin, S.</dc:creator>
<dc:creator>Oputa, O.</dc:creator>
<dc:creator>Martinez, A. M.</dc:creator>
<dc:creator>Nacht, H. D.</dc:creator>
<dc:creator>Tschang, M.</dc:creator>
<dc:creator>Mahmood, M.</dc:creator>
<dc:creator>Tucker, A.</dc:creator>
<dc:creator>Pusuloori, S.</dc:creator>
<dc:creator>Zmoyro, L.</dc:creator>
<dc:creator>Abraira Lab Computational Group,</dc:creator>
<dc:creator>Popovich, P.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>McTigue, D.</dc:creator>
<dc:creator>Tysseling, V. M.</dc:creator>
<dc:creator>Dulin, J.</dc:creator>
<dc:creator>Hollis, E.</dc:creator>
<dc:creator>Datta, S. R.</dc:creator>
<dc:creator>Abraira, V. E. G.</dc:creator>
<dc:date>2023-11-02</dc:date>
<dc:identifier>doi:10.1101/2023.10.31.564826</dc:identifier>
<dc:title><![CDATA[Using behavioral biomarkers to redefine epochs of spontaneous recovery following spinal cord injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.02.565001v1?rss=1">
<title>
<![CDATA[
Feasibility Study Utilizing NanoString Digital Spatial Profiling (DSP) Technology for Characterizing the Immune Microenvironment in Barrett's Esophagus FFPE Tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.02.565001v1?rss=1</link>
<description><![CDATA[
To date, characterization of the Barretts esophagus (BE) immune microenvironment in patients with known progression status to determine how the microenvironment may influence BE progression to esophageal adenocarcinoma (EAC) has been understudied, hindering both the biological understanding of progression and the development of novel diagnostics and therapies. Therefore, this studys aim was to determine if highly multiplex interrogation of the immune microenvironment can be performed on endoscopic formalin-fixed, paraffin-embedded (FFPE) samples utilizing the Nanostring GeoMx digital spatial profiling (GeoMx DSP) platform. We performed spatial proteomic analysis of 49 proteins expressed in the microenvironment and epithelial cells of histologically identical FFPE endoscopic biopsies from patients with non-dysplastic BE (NDBE) who later progressed to high-grade dysplasia (HGD) or EAC (N=7) or from patients who after at least 5 years follow up did not (N=8). In addition, we performed RNA analysis of 1,812 cancer related transcripts on a series of three endoscopic mucosal resections containing regions of normal tissue, BE, dysplasia (DYS), and EAC. Our primary goal was to determine feasibility of this approach and begin to identify the types of specific immune cell populations that may mediate the progression of pre-neoplastic BE to EAC. Spatial proteomic and transcriptomic profiling with GeoMx DSP showed reasonable quality metrics and detected expected differences between epithelium and stroma. Several proteins were found to have increased expression within non-dysplastic BE biopsies from progressors compared to non-progressors, suggesting further studies on the BE microenvironment are warranted.

SummaryNew biological insights into the stepwise development and progression of esophageal adenocarcinoma (EAC) from Barretts esophagus (BE) are imperative to develop tailored approaches for early detection and optimal clinical management of the disease. This study aimed to determine the feasibility to spatially profile stromal and immunologic properties that accompany malignant transformation of BE to EAC in formalin-fixed, paraffin-embedded (FFPE) tissues. NanoStrings Digital Spatial Profiling (DSP) technology can detect and quantify protein and RNA transcripts in a highly multiplexed manner with spatial resolution, within specific regions of interest on FFPE tissue. Here, we performed a pilot study using the Nanostring GeoMx DSP, for measurement of protein and ribonucleic acid (RNA) expression on a series of FFPE slides from endoscopic biopsies and endoscopic mucosal resections (EMR) of BE. We compare a small series of biopsies of non-dysplastic BE (NDBE) from patients who progressed to more advanced disease to patients with NBDE who did not progress and then perform RNA profiling on EMRs with a range of histologic diagnoses.
]]></description>
<dc:creator>Ain, Q. u.</dc:creator>
<dc:creator>Frei, N.</dc:creator>
<dc:creator>Khoshiwal, A. M.</dc:creator>
<dc:creator>Stougie, P.</dc:creator>
<dc:creator>Odze, R.</dc:creator>
<dc:creator>Ferri, L.</dc:creator>
<dc:creator>Duits, L. C.</dc:creator>
<dc:creator>Bergman, J.</dc:creator>
<dc:creator>Stachler, M. D.</dc:creator>
<dc:date>2023-11-02</dc:date>
<dc:identifier>doi:10.1101/2023.11.02.565001</dc:identifier>
<dc:title><![CDATA[Feasibility Study Utilizing NanoString Digital Spatial Profiling (DSP) Technology for Characterizing the Immune Microenvironment in Barrett's Esophagus FFPE Tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.02.565382v1?rss=1">
<title>
<![CDATA[
A diet-dependent host metabolite shapes the gut microbiota to protect from autoimmunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.02.565382v1?rss=1</link>
<description><![CDATA[
Diet can protect from autoimmune disease; however, whether diet acts via the host and/or microbiome remains unclear. Here, we use a ketogenic diet (KD) as a model to dissect these complex interactions. A KD rescued the experimental autoimmune encephalomyelitis (EAE) mouse model of multiple sclerosis in a microbiota-dependent fashion. Dietary supplementation with a single KD-dependent host metabolite ({beta}-hydroxybutyrate, {beta}HB) rescued EAE whereas transgenic mice unable to produce {beta}HB in the intestine developed more severe disease. Transplantation of the {beta}HB-shaped gut microbiota was protective. Lactobacillus sequence variants were associated with decreased T helper 17 (Th17) cell activation in vitro. Finally, we isolated a L. murinus strain that protected from EAE, which was phenocopied by the Lactobacillus metabolite indole lactic acid. Thus, diet alters the immunomodulatory potential of the gut microbiota by shifting host metabolism, emphasizing the utility of taking a more integrative approach to study diet-host-microbiome interactions.
]]></description>
<dc:creator>Alexander, M.</dc:creator>
<dc:creator>Upadhyay, V.</dc:creator>
<dc:creator>Rock, R.</dc:creator>
<dc:creator>Ramirez, L.</dc:creator>
<dc:creator>Puchalska, P.</dc:creator>
<dc:creator>Orellana, D.</dc:creator>
<dc:creator>Ang, Q. Y.</dc:creator>
<dc:creator>Turnbaugh, J. A.</dc:creator>
<dc:creator>Tian, Y.</dc:creator>
<dc:creator>Dumlao, D.</dc:creator>
<dc:creator>Nayak, R. R.</dc:creator>
<dc:creator>Patterson, A. D.</dc:creator>
<dc:creator>Newman, J. C.</dc:creator>
<dc:creator>Crawford, P. A.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:date>2023-11-03</dc:date>
<dc:identifier>doi:10.1101/2023.11.02.565382</dc:identifier>
<dc:title><![CDATA[A diet-dependent host metabolite shapes the gut microbiota to protect from autoimmunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.02.565407v1?rss=1">
<title>
<![CDATA[
Quantitative Three-dimensional Label-free Digital Holographic Imaging of Cardiomyocyte Size, Ploidy, and Cell Division 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.02.565407v1?rss=1</link>
<description><![CDATA[
Cardiac regeneration in newborn rodents depends on the ability of pre-existing cardiomyocytes to proliferate and divide. This capacity is lost within the first week of postnatal development when these cells rapidly switch from hyperplasia to hypertrophy, withdraw from the cell cycle, become binucleated, and increase in size. How these dynamic changes in size and ploidy impact cardiomyocyte proliferative potential is not well understood. In this study, we innovate the application of a commercially available digital holographic imaging microscope, the Holomonitor M4, to evaluate the proliferative responses of mononucleated diploid and binucleated tetraploid cardiomyocytes. This instrument coupled with the powerful Holomonitor App Suite software enables long-term label-free quantitative three-dimensional tracking of primary cardiomyocyte dynamics in real-time with single-cell resolution. Our digital holographic imaging results provide direct evidence that mononucleated cardiomyocytes retain significant proliferative potential as most can successfully divide with high frequency. In contrast, binucleated cardiomyocytes exhibit a blunted response to a proliferative stimulus with the majority not attempting to divide at all. Nevertheless, some binucleated cardiomyocytes were capable of complete division, suggesting that these cells still do retain limited proliferative capacity. By quantitatively tracking cardiomyocyte volume dynamics during these proliferative responses, we reveal that both mononucleated and binucleated cells reach a unique size threshold prior to attempted cell division. The absolute threshold is increased by binucleation, which may limit the ability of binucleated cardiomyocytes to divide. By defining the interrelationship between cardiomyocyte size, ploidy, and cell cycle control, we will better understand the cellular mechanisms that drive the loss of mammalian cardiac regenerative capacity after birth.
]]></description>
<dc:creator>Park, S.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Ross, I.</dc:creator>
<dc:creator>Moreno, J.</dc:creator>
<dc:creator>Khyeam, S.</dc:creator>
<dc:creator>Simmons, J.</dc:creator>
<dc:creator>Huang, G. N.</dc:creator>
<dc:creator>Payumo, A. Y.</dc:creator>
<dc:date>2023-11-04</dc:date>
<dc:identifier>doi:10.1101/2023.11.02.565407</dc:identifier>
<dc:title><![CDATA[Quantitative Three-dimensional Label-free Digital Holographic Imaging of Cardiomyocyte Size, Ploidy, and Cell Division]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.03.565283v1?rss=1">
<title>
<![CDATA[
MET variants with activating N-lobe mutations identified in hereditary papillary renal cell carcinomas still require ligand stimulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.03.565283v1?rss=1</link>
<description><![CDATA[
In hereditary papillary renal cell carcinoma (HPRCC), the hepatocyte growth factor receptor (MET) receptor tyrosine kinase (RTK) mutations recorded to date are located in the kinase domain and lead to constitutive MET activation. This contrasts with MET mutations identified in non-small cell lung cancer (NSCLC), which lead to exon 14 skipping and deletion of a regulatory domain: in this latter case, the mutated receptor still requires ligand stimulation. Sequencing of MET in samples from 158 HPRCC and 2808 NSCLC patients revealed ten uncharacterized mutations. Four of these, all found in HPRCC and leading to amino acid substitutions in the N-lobe of the MET kinase, proved able to induce cell transformation, which was further enhanced by hepatocyte growth factor (HGF) stimulation: His1086Leu, Ile1102Thr, Leu1130Ser and Cis1125Gly. Similar to the variant resulting in MET exon 14 skipping, the two N-lobe MET variants His1086Leu and Ile1102Thr were found to require stimulation by HGF in order to strongly activate downstream signaling pathways and epithelial cell motility. The Ile1102Thr mutation also displayed transforming potential, promoting tumor growth in a xenograft model. In addition, the N-lobe-mutated MET variants were found to trigger a common HGF-stimulation-dependent transcriptional program, consistent with an observed increase in cell motility and invasion. Altogether, this functional characterization revealed that N-lobe variants still require ligand stimulation, in contrast to other RTK variants. This suggests that HGF expression in the tumor microenvironment is important for tumor growth. The sensitivity of these variants to MET inhibitors opens the way for use of targeted therapies for patients harboring the corresponding mutations.
]]></description>
<dc:creator>Guerin, C. M.</dc:creator>
<dc:creator>Vinchent, A.</dc:creator>
<dc:creator>Damour, I.</dc:creator>
<dc:creator>Laratte, A.</dc:creator>
<dc:creator>Tellier, R.</dc:creator>
<dc:creator>Estevam, G.</dc:creator>
<dc:creator>Meneboo, J.-P.</dc:creator>
<dc:creator>Villenet, C.</dc:creator>
<dc:creator>Descarpentries, C.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Figeac, M.</dc:creator>
<dc:creator>Cortot, A. B.</dc:creator>
<dc:creator>Rouleau, E.</dc:creator>
<dc:creator>Tulasne, D.</dc:creator>
<dc:date>2023-11-05</dc:date>
<dc:identifier>doi:10.1101/2023.11.03.565283</dc:identifier>
<dc:title><![CDATA[MET variants with activating N-lobe mutations identified in hereditary papillary renal cell carcinomas still require ligand stimulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.03.565556v1?rss=1">
<title>
<![CDATA[
SuPreMo: a computational tool for streamlining in silico perturbation using sequence-based predictive models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.03.565556v1?rss=1</link>
<description><![CDATA[
Computationally editing genome sequences is a common bioinformatics task, but current approaches have limitations, such as incompatibility with structural variants, challenges in identifying responsible sequence perturbations, and the need for vcf file inputs and phased data. To address these bottlenecks, we present Sequence Mutator for Predictive Models (SuPreMo), a scalable and comprehensive tool for performing in silico mutagenesis. We then demonstrate how pairs of reference and perturbed sequences can be used with machine learning models to prioritize pathogenic variants or discover new functional sequences.

Availability and ImplementationSuPreMo was written in Python, and can be run using only one line of code to generate both sequences and 3D genome disruption scores. The codebase, instructions for installation and use, and tutorials are on the Github page: https://github.com/ketringjoni/SuPreMo/tree/main.

Contactkatherine.pollard@gladstone.ucsf.edu

Supplementary informationSupplementary data are available at Bioinformatics online.
]]></description>
<dc:creator>Gjoni, K.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2023-11-05</dc:date>
<dc:identifier>doi:10.1101/2023.11.03.565556</dc:identifier>
<dc:title><![CDATA[SuPreMo: a computational tool for streamlining in silico perturbation using sequence-based predictive models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.05.565687v1?rss=1">
<title>
<![CDATA[
Contexts facilitate dynamic value encoding in the mesolimbic dopamine system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.05.565687v1?rss=1</link>
<description><![CDATA[
Adaptive behavior in a dynamic environment often requires rapid revaluation of stimuli that deviates from well-learned associations. The divergence between stable value-encoding and appropriate behavioral output remains a critical test to theories of dopamines function in learning, motivation, and motor control. Yet how dopamine neurons are involved in the revaluation of cues when the world changes to alter our behavior remains unclear. Here we make use of pharmacology, in vivo electrophysiology, fiber photometry, and optogenetics to resolve the contributions of the mesolimbic dopamine system to the dynamic reorganization of reward-seeking. Male and female rats were trained to discriminate when a conditioned stimulus would be followed by sucrose reward by exploiting the prior, non-overlapping presentation of a separate discrete cue - an occasion setter. Only when the occasion setters presentation preceded the conditioned stimulus did the conditioned stimulus predict sucrose delivery. As a result, in this task we were able to dissociate the average value of the conditioned stimulus from its immediate expected value on a trial-to-trial basis. Both the activity of ventral tegmental area dopamine neurons and dopamine signaling in the nucleus accumbens were essential for rats to successfully update behavioral responding in response to the occasion setter. Moreover, dopamine release in the nucleus accumbens following the conditioned stimulus only occurred when the occasion setter indicated it would predict reward. Downstream of dopamine release, we found that single neurons in the nucleus accumbens dynamically tracked the value of the conditioned stimulus. Together these results reveal a novel mechanism within the mesolimbic dopamine system for the rapid revaluation of motivation.
]]></description>
<dc:creator>Fraser, K. M.</dc:creator>
<dc:creator>Collins, V. L.</dc:creator>
<dc:creator>Wolff, A. R.</dc:creator>
<dc:creator>Ottenheimer, D. J.</dc:creator>
<dc:creator>Bornhoft, K. N.</dc:creator>
<dc:creator>Pat, F.</dc:creator>
<dc:creator>Chen, B. J.</dc:creator>
<dc:creator>Janak, P. H.</dc:creator>
<dc:creator>Saunders, B. T.</dc:creator>
<dc:date>2023-11-05</dc:date>
<dc:identifier>doi:10.1101/2023.11.05.565687</dc:identifier>
<dc:title><![CDATA[Contexts facilitate dynamic value encoding in the mesolimbic dopamine system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.03.565292v1?rss=1">
<title>
<![CDATA[
Integrated longitudinal multi-omics study identifies immune programs associated with COVID-19 severity and mortality in 1152 hospitalized participants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.03.565292v1?rss=1</link>
<description><![CDATA[
Hospitalized COVID-19 patients exhibit diverse clinical outcomes, with some individuals diverging over time even though their initial disease severity appears similar. A systematic evaluation of molecular and cellular profiles over the full disease course can link immune programs and their coordination with progression heterogeneity. In this study, we carried out deep immunophenotyping and conducted longitudinal multi-omics modeling integrating ten distinct assays on a total of 1,152 IMPACC participants and identified several immune cascades that were significant drivers of differential clinical outcomes. Increasing disease severity was driven by a temporal pattern that began with the early upregulation of immunosuppressive metabolites and then elevated levels of inflammatory cytokines, signatures of coagulation, NETosis, and T-cell functional dysregulation. A second immune cascade, predictive of 28-day mortality among critically ill patients, was characterized by reduced total plasma immunoglobulins and B cells, as well as dysregulated IFN responsiveness. We demonstrated that the balance disruption between IFN-stimulated genes and IFN inhibitors is a crucial biomarker of COVID-19 mortality, potentially contributing to the failure of viral clearance in patients with fatal illness. Our longitudinal multi-omics profiling study revealed novel temporal coordination across diverse omics that potentially explain disease progression, providing insights that inform the targeted development of therapies for hospitalized COVID-19 patients, especially those critically ill.
]]></description>
<dc:creator>Gygi, J. P.</dc:creator>
<dc:creator>Maguire, C.</dc:creator>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Shinde, P.</dc:creator>
<dc:creator>Konstorum, A.</dc:creator>
<dc:creator>Shannon, C. P.</dc:creator>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Hoch, A.</dc:creator>
<dc:creator>Jayavelu, N.</dc:creator>
<dc:creator>IMPACC Network,</dc:creator>
<dc:creator>Haddad, E. K.</dc:creator>
<dc:creator>Reed, E. F.</dc:creator>
<dc:creator>Kraft, M.</dc:creator>
<dc:creator>McComsey, G. A.</dc:creator>
<dc:creator>Metcalf, J.</dc:creator>
<dc:creator>Ozonoff, A.</dc:creator>
<dc:creator>Esserman, D.</dc:creator>
<dc:creator>Cairns, C. B.</dc:creator>
<dc:creator>Rouphael, N.</dc:creator>
<dc:creator>Bosinger, S. E.</dc:creator>
<dc:creator>Kim-Schulze, S.</dc:creator>
<dc:creator>Krammer, F.</dc:creator>
<dc:creator>Rosen, L. B.</dc:creator>
<dc:creator>Bakel, H.</dc:creator>
<dc:creator>Wilson, M.</dc:creator>
<dc:creator>Eckalbar, W.</dc:creator>
<dc:creator>Maecker, H.</dc:creator>
<dc:creator>Langelier, C. R.</dc:creator>
<dc:creator>Steen, H.</dc:creator>
<dc:creator>Altman, M. C.</dc:creator>
<dc:creator>Montgomery, R. R.</dc:creator>
<dc:creator>Levy, O.</dc:creator>
<dc:creator>Melamed, E.</dc:creator>
<dc:creator>Pulendran, B.</dc:creator>
<dc:creator>Arce, J.</dc:creator>
<dc:creator>Smolen, K. K.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Becker, P. M.</dc:creator>
<dc:creator>Augustine, A. D.</dc:creator>
<dc:creator>Sekaly, R. P.</dc:creator>
<dc:creator>Ehrlich, L. I. R.</dc:creator>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Pete</dc:creator>
<dc:date>2023-11-06</dc:date>
<dc:identifier>doi:10.1101/2023.11.03.565292</dc:identifier>
<dc:title><![CDATA[Integrated longitudinal multi-omics study identifies immune programs associated with COVID-19 severity and mortality in 1152 hospitalized participants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.06.565375v1?rss=1">
<title>
<![CDATA[
Single cell transcriptional profiling of benign prostatic hyperplasia reveals a progenitor-like luminal epithelial cell state within an inflammatory microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.06.565375v1?rss=1</link>
<description><![CDATA[
Benign prostatic hyperplasia (BPH) is characterized by excessive cell proliferation and inflammation and affects most aging men. The development of new therapies for BPH requires a deeper understanding of the underlying pathophysiology and cellular components of BPH. Here, we characterize at single cell resolution the cellular states of BPH and identify cell populations enriched in BPH that contribute to cell proliferation and inflammation. Single-cell RNA-sequencing was performed on prostate tissue from 15 patients undergoing holmium laser enucleation of the prostate for treatment of BPH. Clustering and differential expression analysis on aligned single cell RNA-seq data was performed to annotate all cell types. Pseudotime, gene set enrichment, gene ontology, and ligand-receptor analyses were performed. 16,234 cells were analyzed and specific stromal, epithelial, and immune subgroups were found to be strongly associated with inflammation. A rare luminal subgroup was identified and pseudotime analysis indicated this luminal subgroup was more closely related to club and basal cells. Using a gene set derived from epithelial stem cells, we found that this luminal subgroup had a significantly higher stem cell signature score than all other epithelial subgroups, suggesting this subgroup is a luminal precursor state. Ligand-receptor interactions between stromal, epithelial, and immune cells were explored with CellPhoneDB. Unique interactions highlighting MIF, a pro-inflammatory cytokine that promotes epithelial cell growth and inflammation in the prostate, were found between fibroblasts and the progenitor luminal subgroup. This luminal subgroup also interacted with neutrophils and macrophages through MIF. Our single-cell profiling of BPH provides a roadmap for inflammation-linked cell subgroups and highlights a novel luminal progenitor subgroup interacting with other cell groups via MIF that may contribute to the inflammation and cell proliferation phenotype associated with BPH.
]]></description>
<dc:creator>Unno, R.</dc:creator>
<dc:creator>Akutagawa, J.</dc:creator>
<dc:creator>Song, H.</dc:creator>
<dc:creator>Hui, K.</dc:creator>
<dc:creator>Chen, Y.-A.</dc:creator>
<dc:creator>Pham, J.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Huang, F. W.</dc:creator>
<dc:creator>Chi, T.</dc:creator>
<dc:date>2023-11-07</dc:date>
<dc:identifier>doi:10.1101/2023.11.06.565375</dc:identifier>
<dc:title><![CDATA[Single cell transcriptional profiling of benign prostatic hyperplasia reveals a progenitor-like luminal epithelial cell state within an inflammatory microenvironment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.06.565810v1?rss=1">
<title>
<![CDATA[
Gene Expression Meta-Analysis Reveals Aging and Cellular Senescence Signatures in Scleroderma-associated Interstitial Lung Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.06.565810v1?rss=1</link>
<description><![CDATA[
Aging and cellular senescence are increasingly recognized as key contributors to pulmonary fibrosis. However, our understanding in the context of scleroderma associated interstitial lung disease (SSc-ILD) is limited. To investigate, we leveraged previously established lung aging and cell-specific senescence signatures to determine their presence and potential relevance to SSc-ILD. We performed a gene expression meta-analysis of lung tissue from 38 SSc-ILD and 18 healthy controls and found markers (GDF15, COMP, CDKN2A) and pathways (p53) of senescence were significantly increased in SSc-ILD. When probing the established aging and cellular senescence signatures, we found epithelial and fibroblast senescence signatures had a 3.6-fold and 3.7-fold enrichment respectively in the lung tissue of SSc-ILD and that lung aging genes (CDKN2A, FRZB, PDE1A, NAPI12) were increased in SSc-ILD. These signatures were also enriched in SSc skin and associated with degree of skin involvement (limited vs. diffuse cutaneous). To further support these findings, we examined telomere length (TL), a surrogate for aging, in lung tissue and found independent of age, SSc-ILD had significantly shorter telomeres than controls in type II alveolar cells in the lung. TL in SSc-ILD was comparable to idiopathic pulmonary fibrosis, a disease of known aberrant aging. Taken together, this study provides novel insight into the possible mechanistic effects of accelerated aging and aberrant cellular senescence in SSc-ILD pathogenesis.
]]></description>
<dc:creator>Yang, M. M.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Neely, J.</dc:creator>
<dc:creator>Hinchcliff, M.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2023-11-07</dc:date>
<dc:identifier>doi:10.1101/2023.11.06.565810</dc:identifier>
<dc:title><![CDATA[Gene Expression Meta-Analysis Reveals Aging and Cellular Senescence Signatures in Scleroderma-associated Interstitial Lung Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.06.565933v1?rss=1">
<title>
<![CDATA[
TBXT dose sensitivity and the decoupling of nascent mesoderm specification from EMT progression in 2D human gastruloids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.06.565933v1?rss=1</link>
<description><![CDATA[
In the nascent mesoderm, levels of Brachyury (TBXT) expression must be precisely regulated to ensure cells exit the primitive streak and pattern the anterior-posterior axis, but how this varying dosage informs morphogenesis is not well understood. In this study, we define the transcriptional consequences of TBXT dose reduction during early human gastrulation using human induced pluripotent stem cell (hiPSC)-based models of gastrulation and mesoderm differentiation. Multiomic single-nucleus RNA and single-nucleus ATAC sequencing of 2D gastruloids comprised of WT, TBXT heterozygous (TBXT-Het), or TBXT null (TBXT-KO) hiPSCs reveal that varying TBXT dosage does not compromise a cells ability to differentiate into nascent mesoderm, but that the loss of TBXT significantly delays the temporal progression of the epithelial to mesenchymal transition (EMT). This delay is dependent on TBXT dose, as cells heterozygous for TBXT proceed with EMT at an intermediate pace relative to WT or TBXT-KO. By differentiating iPSCs of the allelic series into nascent mesoderm in a monolayer format, we further illustrate that TBXT dose directly impacts the persistence of junctional proteins and cell-cell adhesions. These results demonstrate that EMT progression can be decoupled from the acquisition of mesodermal identity in the early gastrula and shed light on the mechanisms underlying human embryogenesis.
]]></description>
<dc:creator>Bulger, E. A.</dc:creator>
<dc:creator>Muncie-Vasic, I.</dc:creator>
<dc:creator>Libby, A. R. G.</dc:creator>
<dc:creator>McDevitt, T. C.</dc:creator>
<dc:creator>Bruneau, B. G.</dc:creator>
<dc:date>2023-11-07</dc:date>
<dc:identifier>doi:10.1101/2023.11.06.565933</dc:identifier>
<dc:title><![CDATA[TBXT dose sensitivity and the decoupling of nascent mesoderm specification from EMT progression in 2D human gastruloids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.07.566008v1?rss=1">
<title>
<![CDATA[
The entropic heart: Tracking the psychedelic state via heart rate dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.07.566008v1?rss=1</link>
<description><![CDATA[
A growing body of work shows that autonomic signals provide a privileged evidence-stream to capture various aspects of subjective and neural states. This work investigates the potential for autonomic markers to track the effects of psychedelics -- potent psychoactive drugs with important scientific and clinical value. For this purpose, we introduce a novel Bayesian framework to estimate the entropy of heart rate dynamics under psychedelics. We also calculate Bayesian estimates of mean heart rate and heart rate variability, and investigate how these measures relate to subjective reports and neural effects. Results on datasets covering four drugs -- lysergic acid diethylamide (LSD), dimethyltryptamine (DMT), psilocybin, and sub-anaesthetic doses of the dissociative agent ketamine -- show consistent increases in mean heart rate, high-frequency heart rate variability, and heart rate entropy during the psychedelic experience. Moreover, these effects have predictive power over various dimensions of the psychedelic experience. Changes in heart rate entropy were found to be correlated with increases in brain entropy, while other autonomic markers were not. Overall, our results show that a cost-efficient autonomic measure has the potential to reveal surprising detail about subjective and brain states, opening up a range of new research avenues to explore in both basic and clinical neuroscience.
]]></description>
<dc:creator>Rosas, F. E.</dc:creator>
<dc:creator>Mediano, P. A. M.</dc:creator>
<dc:creator>Timmermann, C.</dc:creator>
<dc:creator>Luppi, A. I.</dc:creator>
<dc:creator>Candia-Rivera, D.</dc:creator>
<dc:creator>Abbasi-Asl, R.</dc:creator>
<dc:creator>Gazzaley, A.</dc:creator>
<dc:creator>Kringelbach, M. L.</dc:creator>
<dc:creator>Muthukumaraswamy, S. D.</dc:creator>
<dc:creator>Bor, D.</dc:creator>
<dc:creator>Garfinkel, S.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:date>2023-11-09</dc:date>
<dc:identifier>doi:10.1101/2023.11.07.566008</dc:identifier>
<dc:title><![CDATA[The entropic heart: Tracking the psychedelic state via heart rate dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.07.564025v1?rss=1">
<title>
<![CDATA[
A novel mouse model of cerebral adrenoleukodystrophy highlights NLRP3 activity in lesion pathogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.07.564025v1?rss=1</link>
<description><![CDATA[
ObjectiveWe sought to create and characterize a mouse model of the inflammatory, cerebral demyelinating phenotype of X-linked adrenoleukodystrophy (ALD) that would facilitate the study of disease pathogenesis and therapy development. We also sought to cross-validate potential therapeutic targets such as fibrin, oxidative stress, and the NLRP3 inflammasome, in post-mortem human and murine brain tissues.

BackgroundALD is caused by mutations in the gene ABCD1 encoding a peroxisomal transporter. More than half of males with an ABCD1 mutation develop the cerebral phenotype (cALD). Incomplete penetrance and absence of a genotype-phenotype correlation imply a role for environmental triggers. Mechanistic studies have been limited by the absence of a cALD phenotype in the Abcd1-null mouse.

MethodsWe generated a cALD phenotype in 8-week-old, male Abcd1-null mice by deploying a two-hit method that combines cuprizone (CPZ) and experimental autoimmune encephalomyelitis (EAE) models. We employed in vivo MRI and post-mortem immunohistochemistry to evaluate myelin loss, astrogliosis, blood-brain barrier (BBB) disruption, immune cell infiltration, fibrin deposition, oxidative stress, and Nlrp3 inflammasome activation in mice. We used bead-based immunoassay and immunohistochemistry to evaluate IL-18 in CSF and post-mortem human cALD brain tissue.

ResultsMRI studies revealed T2 hyperintensities and post-gadolinium enhancement in the medial corpus callosum of cALD mice, similar to human cALD lesions. Both human and mouse cALD lesions shared common histologic features of myelin phagocytosis, myelin loss, abundant microglial activation, T and B-cell infiltration, and astrogliosis. Compared to wild-type controls, Abcd1-null mice had more severe cerebral inflammation, demyelination, fibrin deposition, oxidative stress, and IL-18 activation. IL-18 immunoreactivity co-localized with macrophages/microglia in the perivascular region of both human and mouse brain tissue.

InterpretationThis novel mouse model of cALD suggests loss of Abcd1 function predisposes to more severe cerebral inflammation, oxidative stress, fibrin deposition, and Nlrp3 pathway activation, which parallels the findings seen in humans with cALD. We expect this model to enable long-sought investigations into cALD mechanisms and accelerate development of candidate therapies for lesion prevention, cessation, and remyelination.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/564025v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Hashemi, E.</dc:creator>
<dc:creator>Srivastava, I.</dc:creator>
<dc:creator>Aguirre, A.</dc:creator>
<dc:creator>Kaushal, E.</dc:creator>
<dc:creator>Yoseph, E.</dc:creator>
<dc:creator>Awani, A.</dc:creator>
<dc:creator>Ryu, J. K.</dc:creator>
<dc:creator>Akassoglou, K.</dc:creator>
<dc:creator>Talebian, S.</dc:creator>
<dc:creator>Chu, P.</dc:creator>
<dc:creator>Pisani, L.</dc:creator>
<dc:creator>Musolino, P.</dc:creator>
<dc:creator>Steinman, L.</dc:creator>
<dc:creator>Doyle, K.</dc:creator>
<dc:creator>Robinson, W. H.</dc:creator>
<dc:creator>Sharpe, O.</dc:creator>
<dc:creator>Cayrol, R.</dc:creator>
<dc:creator>Orchard, P.</dc:creator>
<dc:creator>Lund, T.</dc:creator>
<dc:creator>Vogel, H.</dc:creator>
<dc:creator>Lenail, M.</dc:creator>
<dc:creator>Han, M.</dc:creator>
<dc:creator>Bonkowsky, J. P.</dc:creator>
<dc:creator>Van Haren, K. P.</dc:creator>
<dc:date>2023-11-10</dc:date>
<dc:identifier>doi:10.1101/2023.11.07.564025</dc:identifier>
<dc:title><![CDATA[A novel mouse model of cerebral adrenoleukodystrophy highlights NLRP3 activity in lesion pathogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.07.566033v1?rss=1">
<title>
<![CDATA[
Coordinated immune dysregulation in Juvenile Dermatomyositis revealed by single-cell genomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.07.566033v1?rss=1</link>
<description><![CDATA[
Juvenile Dermatomyositis (JDM) is one of several childhood-onset autoimmune disorders characterized by a type I interferon response and autoantibodies. Treatment options are limited due to incomplete understanding of how the disease emerges from dysregulated cell states across the immune system. We therefore investigated the blood of JDM patients at different stages of disease activity using single-cell transcriptomics paired with surface protein expression. By immunophenotyping peripheral blood mononuclear cells, we observed skewing of the B cell compartment towards an immature naive state as a hallmark of JDM. Furthermore, we find that these changes in B cells are paralleled by signatures of Th2-mediated inflammation. Additionally, our work identified SIGLEC-1 expression in monocytes as a composite measure of heterogeneous type I interferon activity in disease. We applied network analysis to reveal that hyperactivation of the type I interferon response in all immune populations is coordinated with dysfunctional protein processing and regulation of cell death programming. This analysis separated the ubiquitously expressed type I interferon response into a central hub and revealed previously masked cell states. Together, these findings reveal the coordinated immune dysregulation underpinning JDM and provide novel insight into strategies for restoring balance in immune function.
]]></description>
<dc:creator>Rabadam, G.</dc:creator>
<dc:creator>Wibrand, C.</dc:creator>
<dc:creator>Flynn, E.</dc:creator>
<dc:creator>Hartoularos, G. C.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Neely, J.</dc:creator>
<dc:date>2023-11-10</dc:date>
<dc:identifier>doi:10.1101/2023.11.07.566033</dc:identifier>
<dc:title><![CDATA[Coordinated immune dysregulation in Juvenile Dermatomyositis revealed by single-cell genomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.09.566272v1?rss=1">
<title>
<![CDATA[
Light induced expression of gRNA allows for optogenetic gene editing of T lymphocytes in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.09.566272v1?rss=1</link>
<description><![CDATA[
There is currently a lack of tools capable of perturbing genes in both a precise and spatiotemporal fashion. CRISPRs ease of use and flexibility, coupled with lights unparalleled spatiotemporal resolution deliverable from a controllable source, makes optogenetic CRISPR a well-suited solution for precise spatiotemporal gene perturbations. Here we present a new optogenetic CRISPR tool, BLU-VIPR, that diverges from prevailing split-Cas design strategies and instead focuses on optogenetic regulation of gRNA production. This simplifies spatiotemporal gene perturbation and works in vivo with cells previously intractable to optogenetic gene editing. We engineered BLU-VIPR around a new potent blue-light activated transcription factor and ribozyme-flanked gRNA. The BLU-VIPR design is genetically encoded and ensures precise excision of multiple gRNAs from a single mRNA transcript, allowing for optogenetic gene editing in T lymphocytes in vivo.
]]></description>
<dc:creator>Velasquez Pulgarin, D. A.</dc:creator>
<dc:creator>Pelo, N.</dc:creator>
<dc:creator>Ferrandiz, L.</dc:creator>
<dc:creator>Trselic, T.</dc:creator>
<dc:creator>Nyberg, W. A.</dc:creator>
<dc:creator>Bowlin, G. L.</dc:creator>
<dc:creator>Espinosa, A.</dc:creator>
<dc:date>2023-11-10</dc:date>
<dc:identifier>doi:10.1101/2023.11.09.566272</dc:identifier>
<dc:title><![CDATA[Light induced expression of gRNA allows for optogenetic gene editing of T lymphocytes in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.07.565473v1?rss=1">
<title>
<![CDATA[
A high-density diffuse optical tomography dataset of naturalistic viewing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.07.565473v1?rss=1</link>
<description><![CDATA[
Traditional laboratory tasks offer tight experimental control but lack the richness of our everyday human experience. As a result, many cognitive neuroscientists have been motivated to adopt experimental paradigms that are more natural, such as stories and movies. Here we describe data collected from 58 healthy adult participants (aged 18-76 years) who viewed 10 minutes of a movie (The Good, the Bad, and the Ugly, 1966). Most (36) participants viewed the clip more than once, resulting in 106 sessions of data. Cortical responses were mapped using high-density diffuse optical tomography (first-through fourth nearest neighbor separations of 1.3, 3.0, 3.9, and 4.7 cm), covering large portions of superficial occipital, temporal, parietal, and frontal lobes. Consistency of measured activity across subjects was quantified using intersubject correlation analysis. Data are provided in both channel format (SNIRF) and projected to standard space (NIfTI) using an atlas-based light model. These data are suitable for methods exploration as well as investigating a wide variety of cognitive phenomena.
]]></description>
<dc:creator>Sherafati, A.</dc:creator>
<dc:creator>Bajracharya, A.</dc:creator>
<dc:creator>Jones, M.</dc:creator>
<dc:creator>Speh, E.</dc:creator>
<dc:creator>Munsi, M.</dc:creator>
<dc:creator>Lin, C.-H. P.</dc:creator>
<dc:creator>Fishell, A. K.</dc:creator>
<dc:creator>Hershey, T.</dc:creator>
<dc:creator>Eggebrecht, A. T.</dc:creator>
<dc:creator>Culver, J. P.</dc:creator>
<dc:creator>Peelle, J. E.</dc:creator>
<dc:date>2023-11-11</dc:date>
<dc:identifier>doi:10.1101/2023.11.07.565473</dc:identifier>
<dc:title><![CDATA[A high-density diffuse optical tomography dataset of naturalistic viewing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.11.566697v1?rss=1">
<title>
<![CDATA[
Deletion of sulfate transporter SUL1 extends yeast replicative lifespan via reduced PKA signaling instead of decreased sulfate uptake 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.11.566697v1?rss=1</link>
<description><![CDATA[
The regulation of cellular metabolism and growth in response to nutrient availability is crucial for cell survival and can significantly impact on lifespan. Central to this regulation is a class of transporters that sense and transport specific nutrients and transduce the signal downstream to control genes responsible for growth and survival. In this study, we identified SUL1, a plasma membrane transporter responsible for regulating the entry of extracellular sulfate in Saccharomyces cerevisiae, as a key gene for regulating lifespan. We conducted a systematic analysis to delineate the downstream mechanism underlying the lifespan extension by SUL1 deletion. Surprisingly, we found that the lifespan extending effect of SUL1 deletion is not due to decreased sulfate transport. The SUL1 deletion mutant exhibited decreased PKA signaling, resulting in a series of downstream effects, including increased stress-protective trehalose and glycogen, increased nuclear translocation of MSN2, elevated expression of general stress response genes, enhanced autophagy, and reduced expression of amino acid biosynthetic and ribosomal genes. We demonstrated that the observed increase in lifespan is dependent on MSN2 and autophagy pathways. Our findings exemplify the influence of nutrient signaling rather than the nutrient itself on lifespan regulation and further substantiate the pivotal role of the PKA pathway in this process.
]]></description>
<dc:creator>Long, J.</dc:creator>
<dc:creator>Ma, M.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Gong, B.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:date>2023-11-11</dc:date>
<dc:identifier>doi:10.1101/2023.11.11.566697</dc:identifier>
<dc:title><![CDATA[Deletion of sulfate transporter SUL1 extends yeast replicative lifespan via reduced PKA signaling instead of decreased sulfate uptake]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.10.566510v1?rss=1">
<title>
<![CDATA[
Microglia Depletion Reduces Human Neuronal APOE4-Driven Pathologies in a Chimeric Alzheimer's Disease Model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.10.566510v1?rss=1</link>
<description><![CDATA[
Despite strong evidence supporting the involvement of both apolipoprotein E4 (APOE4) and microglia in Alzheimers Disease (AD) pathogenesis, the effects of microglia on neuronal APOE4-driven AD pathogenesis remain elusive. Here, we examined such effects utilizing microglial depletion in a chimeric model with human neurons in mouse hippocampus. Specifically, we transplanted homozygous APOE4, isogenic APOE3, and APOE-knockout (APOE-KO) induced pluripotent stem cell (iPSC)-derived human neurons into the hippocampus of human APOE3 or APOE4 knock-in mice, and depleted microglia in half the chimeric mice. We found that both neuronal APOE and microglial presence were important for the formation of A{beta} and tau pathologies in an APOE isoform-dependent manner (APOE4 > APOE3). Single-cell RNA-sequencing analysis identified two pro-inflammatory microglial subtypes with high MHC-II gene expression that are enriched in chimeric mice with human APOE4 neuron transplants. These findings highlight the concerted roles of neuronal APOE, especially APOE4, and microglia in AD pathogenesis.

HIGHLIGHTSO_LITransplanted human APOE4 neurons generate A{beta} and p-tau aggregates in APOE4-KI mouse hippocampus.
C_LIO_LIHuman neuronal APOE4 promotes the formation of dense-core A{beta} plaques and p-tau aggregates.
C_LIO_LIMicroglia is required for human neuronal APOE4-driven formation of p-tau aggregates.
C_LIO_LIscRNA-seq reveals enrichment of MHC-II microglia in mice with human APOE4 neuron transplants.
C_LI
]]></description>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Chen, N.</dc:creator>
<dc:creator>Kim, M. J.</dc:creator>
<dc:creator>Blumenfeld, J.</dc:creator>
<dc:creator>Yip, O.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Liang, Z.</dc:creator>
<dc:creator>Nelson, M. R.</dc:creator>
<dc:creator>Koutsodendris, N.</dc:creator>
<dc:creator>Grone, B.</dc:creator>
<dc:creator>Ding, L.</dc:creator>
<dc:creator>Yoon, S. Y.</dc:creator>
<dc:creator>Arriola, P.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:date>2023-11-14</dc:date>
<dc:identifier>doi:10.1101/2023.11.10.566510</dc:identifier>
<dc:title><![CDATA[Microglia Depletion Reduces Human Neuronal APOE4-Driven Pathologies in a Chimeric Alzheimer's Disease Model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.11.566649v1?rss=1">
<title>
<![CDATA[
Proteome-wide base editor screens to assess phosphorylation site functionality in high-throughput 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.11.566649v1?rss=1</link>
<description><![CDATA[
Signaling pathways that drive gene expression are typically depicted as having a dozen or so landmark phosphorylation and transcriptional events. In reality, thousands of dynamic post-translational modifications (PTMs) orchestrate nearly every cellular function, and we lack technologies to find causal links between these vast biochemical pathways and genetic circuits at scale. Here, we describe "signaling-to-transcription network" mapping through the development of PTM-centric base editing coupled to phenotypic screens, directed by temporally-resolved phosphoproteomics. Using T cell activation as a model, we observe hundreds of unstudied phosphorylation sites that modulate NFAT transcriptional activity. We identify the phosphorylation-mediated nuclear localization of the phosphatase PHLPP1 which promotes NFAT but inhibits NF{kappa}B activity. We also find that specific phosphosite mutants can alter gene expression in subtle yet distinct patterns, demonstrating the potential for fine-tuning transcriptional responses. Overall, base editor screening of PTM sites provides a powerful platform to dissect PTM function within signaling pathways.
]]></description>
<dc:creator>Kennedy, P. H.</dc:creator>
<dc:creator>Deh Sheikh, A. A.</dc:creator>
<dc:creator>Balakar, M.</dc:creator>
<dc:creator>Jones, A. C.</dc:creator>
<dc:creator>Olive, M. E.</dc:creator>
<dc:creator>Hedge, M.</dc:creator>
<dc:creator>Matias, M. I.</dc:creator>
<dc:creator>Pirete, N.</dc:creator>
<dc:creator>Burt, R.</dc:creator>
<dc:creator>Levy, J.</dc:creator>
<dc:creator>Little, T.</dc:creator>
<dc:creator>Hogan, P. G.</dc:creator>
<dc:creator>Liu, D. R.</dc:creator>
<dc:creator>Doench, J. G.</dc:creator>
<dc:creator>Newton, A. C.</dc:creator>
<dc:creator>Gottschalk, R. A.</dc:creator>
<dc:creator>de Boer, C.</dc:creator>
<dc:creator>Alarcon, S.</dc:creator>
<dc:creator>Newby, G.</dc:creator>
<dc:creator>Myers, S. A.</dc:creator>
<dc:date>2023-11-14</dc:date>
<dc:identifier>doi:10.1101/2023.11.11.566649</dc:identifier>
<dc:title><![CDATA[Proteome-wide base editor screens to assess phosphorylation site functionality in high-throughput]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.10.564562v1?rss=1">
<title>
<![CDATA[
Lipid metabolism drives allele-specific early-stage hypertrophic cardiomyopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.10.564562v1?rss=1</link>
<description><![CDATA[
Hypertrophic cardiomyopathy (HCM) results from pathogenic variants in sarcomeric protein genes, that increase myocyte energy demand and lead to cardiac hypertrophy. But it is unknown whether a common metabolic trait underlies the cardiac phenotype at early disease stage. This study characterized two HCM mouse models (R92W-TnT, R403Q-MyHC) that demonstrate differences in mitochondrial function at early disease stage. Using a combination of cardiac phenotyping, transcriptomics, mass spectrometry-based metabolomics and computational modeling, we discovered allele-specific differences in cardiac structure/function and metabolic changes. TnT-mutant hearts had impaired energy substrate metabolism and increased phospholipid remodeling compared to MyHC-mutants. TnT-mutants showed increased incorporation of saturated fatty acid residues into ceramides, cardiolipin, and increased lipid peroxidation, that could underlie allele-specific differences in mitochondrial function and cardiomyopathy.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=172 HEIGHT=200 SRC="FIGDIR/small/564562v1_ufig1.gif" ALT="Figure 1">
O_LINKSMALLFIG WIDTH=185 HEIGHT=200 SRC="FIGDIR/small/564562v1_ufig2.gif" ALT="Figure 1">
O_LINKSMALLFIG WIDTH=200 HEIGHT=74 SRC="FIGDIR/small/564562v1_ufig3.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@195b1d7org.highwire.dtl.DTLVardef@cead88org.highwire.dtl.DTLVardef@e2bf35org.highwire.dtl.DTLVardef@776765_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Vaniya, A.</dc:creator>
<dc:creator>Karlstaedt, A.</dc:creator>
<dc:creator>Ates Gulkok, D.</dc:creator>
<dc:creator>Thottakara, T.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Fan, S.</dc:creator>
<dc:creator>Eades, H.</dc:creator>
<dc:creator>Fukunaga, R.</dc:creator>
<dc:creator>Vernon, H. J.</dc:creator>
<dc:creator>Fiehn, O.</dc:creator>
<dc:creator>Abraham, M. R.</dc:creator>
<dc:date>2023-11-15</dc:date>
<dc:identifier>doi:10.1101/2023.11.10.564562</dc:identifier>
<dc:title><![CDATA[Lipid metabolism drives allele-specific early-stage hypertrophic cardiomyopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.11.566723v1?rss=1">
<title>
<![CDATA[
Testing of putative antiseizure drugs in a preclinical Dravet syndrome zebrafish model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.11.566723v1?rss=1</link>
<description><![CDATA[
Dravet syndrome (DS) is a severe genetic epilepsy primarily caused by de novo mutations in a voltage-activated sodium channel gene (SCN1A). Patients face life-threatening seizures that are largely resistant to available anti-seizure medications (ASM). Preclinical DS animal models are a valuable tool to identify candidate ASMs for these patients. Among these, scn1lab mutant zebrafish exhibiting spontaneous seizure-like activity are particularly amenable to large-scale drug screening. Prior screening in a scn1lab mutant zebrafish line generated using N-ethyl-N-nitrosourea (ENU) identified valproate, stiripentol, and fenfluramine e.g., Federal Drug Administration (FDA) approved drugs with clinical application in the DS population. Successful phenotypic screening in scn1lab mutant zebrafish consists of two stages: (i) a locomotion-based assay measuring high-velocity convulsive swim behavior and (ii) an electrophysiology-based assay, using in vivo local field potential (LFP) recordings, to quantify electrographic seizure-like events. Using this strategy more than 3000 drug candidates have been screened in scn1lab zebrafish mutants. Here, we curated a list of nine additional anti-seizure drug candidates recently identified in preclinical models: 1-EBIO, AA43279, chlorzoxazone, donepezil, lisuride, mifepristone, pargyline, soticlestat and vorinostat. First-stage locomotion-based assays in scn1lab mutant zebrafish identified only 1-EBIO, chlorzoxazone and lisuride. However, second-stage LFP recording assays did not show significant suppression of spontaneous electrographic seizure activity for any of the nine anti-seizure drug candidates. Surprisingly, soticlestat induced frank electrographic seizure-like discharges in wild-type control zebrafish. Taken together, our results failed to replicate clear anti-seizure efficacy for these drug candidates highlighting a necessity for strict scientific standards in preclinical identification of ASMs.
]]></description>
<dc:creator>Whyte-Fagundes, P.</dc:creator>
<dc:creator>Vance, A.</dc:creator>
<dc:creator>Carroll, A.</dc:creator>
<dc:creator>Figueroa, F.</dc:creator>
<dc:creator>Manukyan, C.</dc:creator>
<dc:creator>Baraban, S. C.</dc:creator>
<dc:date>2023-11-15</dc:date>
<dc:identifier>doi:10.1101/2023.11.11.566723</dc:identifier>
<dc:title><![CDATA[Testing of putative antiseizure drugs in a preclinical Dravet syndrome zebrafish model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.13.566775v1?rss=1">
<title>
<![CDATA[
Enhancing CAR-T Cell Metabolism to Overcome Hypoxic Conditions in the Brain Tumor Microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.13.566775v1?rss=1</link>
<description><![CDATA[
The efficacy of chimeric antigen receptor (CAR)-T therapy has been limited against brain tumors to date. CAR-T cells infiltrating syngeneic intracerebral SB28-EGFRvIII glioma revealed impaired mitochondrial ATP production and a markedly hypoxic status compared to ones migrating to subcutaneous tumors. Drug screenings to improve metabolic states of T cells under hypoxic conditions led us to evaluate the combination of AMPK activator Metformin and the mTOR inhibitor Rapamycin (Met+Rap). Met+Rap-pretreated mouse CAR-T cells showed activated PPAR-gamma coactivator 1 (PGC-1) through mTOR inhibition and AMPK activation, and a higher level of mitochondrial spare respiratory capacity than those pretreated with individual drugs or without pretreatment. Moreover, Met+Rap-pretreated CAR-T cells demonstrated persistent and effective anti-glioma cytotoxic activities in the hypoxic condition. Furthermore, a single intravenous infusion of Met+Rap-pretreated CAR-T cells significantly extended the survival of mice bearing intracerebral SB28-EGFRvIII gliomas. Mass cytometric analyses highlighted increased glioma-infiltrating CAR-T cells in the Met+Rap group with fewer Ly6c+ CD11b+ monocytic myeloid-derived suppressor cells in the tumors. Finally, human CAR-T cells pretreated with Met+Rap recapitulated the observations with murine CAR-T cells, demonstrating improved functions in vitro hypoxic conditions. These findings advocate for translational and clinical exploration of Met+Rap-pretreated CAR-T cells in human trials.
]]></description>
<dc:creator>Hatae, R.</dc:creator>
<dc:creator>Kyewalabye, K.</dc:creator>
<dc:creator>Yamamichi, A.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Phyu, S.</dc:creator>
<dc:creator>Chuntova, P.</dc:creator>
<dc:creator>Nejo, T.</dc:creator>
<dc:creator>Levine, L. S.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:creator>Okada, H.</dc:creator>
<dc:date>2023-11-15</dc:date>
<dc:identifier>doi:10.1101/2023.11.13.566775</dc:identifier>
<dc:title><![CDATA[Enhancing CAR-T Cell Metabolism to Overcome Hypoxic Conditions in the Brain Tumor Microenvironment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.13.566764v1?rss=1">
<title>
<![CDATA[
A Photoactivatable Norepinephrine for Probing Adrenergic Neural Circuits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.13.566764v1?rss=1</link>
<description><![CDATA[
Norepinephrine (NE) is a critical neuromodulator that mediates a wide range of behavior and neurophysiology, including attention, arousal, plasticity, and memory consolidation. A major source of NE is the brainstem nucleus the locus coeruleus (LC), which sends widespread projections throughout the central nervous system (CNS). Efforts to dissect this complex noradrenergic circuitry have driven the development of many tools that detect endogenous NE or modulate widespread NE release via LC activation and inhibition. While these tools have enabled research that elucidates physiological roles of NE, additional tools to probe these circuits with a higher degree of spatial precision could enable a finer delineation of function. Here, we describe the synthesis and chemical properties of a photo-activatable NE, [Ru(bpy)2(PMe3)(NE)]PF6 (RuBi-NE). We validate the one-photon (1P) release of NE using whole-cell patch clamp electrophysiology in acute mouse brain slices containing the LC. We show that a 10 ms pulse of blue light, in the presence of RuBi-NE, briefly modulates the firing rate of LC neurons via -2 adrenergic receptors. The development of a photo-activatable NE that can be released with light in the visible spectrum provides a new tool for fine-grained mapping of complex noradrenergic circuits, as well as the ability to probe how NE acts on non-neuronal cells in the CNS.
]]></description>
<dc:creator>Cahill, M. K.</dc:creator>
<dc:creator>Perez, Y. R.</dc:creator>
<dc:creator>Larpthaveesarp, A.</dc:creator>
<dc:creator>Etchenique, R.</dc:creator>
<dc:creator>Poskanzer, K. E.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.13.566764</dc:identifier>
<dc:title><![CDATA[A Photoactivatable Norepinephrine for Probing Adrenergic Neural Circuits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.13.566926v1?rss=1">
<title>
<![CDATA[
kmerDB: A Database Encompassing the Set of Genomic and Proteomic Sequence Information for Each Species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.13.566926v1?rss=1</link>
<description><![CDATA[
The rapid decline in sequencing cost has enabled the generation of reference genomes and proteomes for a growing number of organisms. However, at the present time, there is no established repository that provides information about organism-specific genomic and proteomic sequences of certain lengths, also known as kmers, that are either present or absent in each genome or proteome. In this article, we present kmerDB, a database accessible through an interactive web interface that provides kmer based information from genomic and proteomic sequences in a systematic way. kmerDB currently contains 202,340,859,107 base pairs and 19,304,903,356 amino acids, spanning 45,785 and 22,386 reference genomes and proteomes, respectively, as well as 14,658,776 and 149,264,442 genomic and proteomic species-specific sequences, termed quasi-primes. Additionally, we provide access to 5,186,757 nucleic and 214,904,089 peptide sequences that are absent from every genome and proteome, termed primes. kmerDB features a user-friendly interface offering various search options and filters for easy parsing and searching. The service is available at: www.kmerdb.com.
]]></description>
<dc:creator>Mouratidis, I.</dc:creator>
<dc:creator>Baltoumas, F. A.</dc:creator>
<dc:creator>Chantzi, N.</dc:creator>
<dc:creator>Chan, C.</dc:creator>
<dc:creator>Montgomery, A.</dc:creator>
<dc:creator>Konnaris, M. A.</dc:creator>
<dc:creator>Georgakopoulos, G. C.</dc:creator>
<dc:creator>Das, A.</dc:creator>
<dc:creator>Chartoumpekis, D.</dc:creator>
<dc:creator>Kovac, J.</dc:creator>
<dc:creator>Pavlopoulos, G.</dc:creator>
<dc:creator>Georgakopoulos-Soares, I.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.13.566926</dc:identifier>
<dc:title><![CDATA[kmerDB: A Database Encompassing the Set of Genomic and Proteomic Sequence Information for Each Species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.14.566937v1?rss=1">
<title>
<![CDATA[
Telomerase reverse transcriptase induces targetable alterations in glutathione and nucleotide biosynthesis in glioblastomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.14.566937v1?rss=1</link>
<description><![CDATA[
Telomerase reverse transcriptase (TERT) is essential for glioblastoma (GBM) proliferation. Delineating metabolic vulnerabilities induced by TERT can lead to novel GBM therapies. We previously showed that TERT upregulates glutathione (GSH) pool size in GBMs. Here, we show that TERT acts via the FOXO1 transcription factor to upregulate expression of the catalytic subunit of glutamate-cysteine ligase (GCLC), the rate-limiting enzyme of de novo GSH synthesis. Inhibiting GCLC using siRNA or buthionine sulfoximine (BSO) reduces synthesis of 13C-GSH from [U-13C]-glutamine and inhibits clonogenicity. However, GCLC inhibition does not induce cell death, an effect that is associated with elevated [U-13C]-glutamine metabolism to glutamate and pyrimidine nucleotide biosynthesis. Mechanistically, GCLC inhibition activates MYC and leads to compensatory upregulation of two key glutamine-utilizing enzymes i.e., glutaminase (GLS), which generates glutamate from glutamine, and CAD (carbamoyl-phosphate synthetase 2, aspartate transcarbamoylase, dihydroorotatase), the enzyme that converts glutamine to the pyrimidine nucleotide precursor dihydroorotate. We then examined the therapeutic potential of inhibiting GLS and CAD in combination with GCLC. 6-diazo-5-oxy-L-norleucin (DON) is a potent inhibitor of glutamine-utilizing enzymes including GLS and CAD. The combination of BSO and DON suppresses GSH and pyrimidine nucleotide biosynthesis and is synergistically lethal in GBM cells. Importantly, in vivo stable isotope tracing indicates that combined treatment with JHU-083 (a brain-penetrant prodrug of DON) and BSO abrogates synthesis of GSH and pyrimidine nucleotides from [U-13C]-glutamine and induces tumor shrinkage in mice bearing intracranial GBM xenografts. Collectively, our studies exploit a mechanistic understanding of TERT biology to identify synthetically lethal metabolic vulnerabilities in GBMs.

SIGNIFICANCEUsing in vivo stable isotope tracing, metabolomics, and loss-of-function studies, we demonstrate that TERT expression is associated with metabolic alterations that can be synergistically targeted for therapy in glioblastomas.
]]></description>
<dc:creator>Udutha, S.</dc:creator>
<dc:creator>Taglang, C.</dc:creator>
<dc:creator>Batsios, G.</dc:creator>
<dc:creator>Gillespie, A. M.</dc:creator>
<dc:creator>Tran, M.</dc:creator>
<dc:creator>Ronen, S. M.</dc:creator>
<dc:creator>ten Hoeve, J.</dc:creator>
<dc:creator>Graeber, T. G.</dc:creator>
<dc:creator>Viswanath, P.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.14.566937</dc:identifier>
<dc:title><![CDATA[Telomerase reverse transcriptase induces targetable alterations in glutathione and nucleotide biosynthesis in glioblastomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.14.567090v1?rss=1">
<title>
<![CDATA[
De novo Design of Peptides that Bind Specific Conformers of α-Synuclein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.14.567090v1?rss=1</link>
<description><![CDATA[
Insoluble amyloids rich in cross-{beta} fibrils are observed in a number of neurodegenerative diseases. Depending on the clinicopathology, the amyloids can adopt distinct supramolecular assemblies, termed conformational strains. However, rapid methods to study amyloid in a conformationally specific manner are lacking. We introduce a novel computational method for de novo design of peptides that tile the surface of -synuclein fibrils in a conformationally specific manner. Our method begins by identifying surfaces that are unique to the conformational strain of interest, which becomes a "target backbone" for the design of a peptide binder. Next, we interrogate structures in the PDB database with high geometric complementarity to the target. Then, we identify secondary structural motifs that interact with this target backbone in a favorable, highly occurring geometry. This method produces monomeric helical motifs with a favorable geometry for interaction with the strands of the underlying amyloid. Each motif is then symmetrically replicated to form a monolayer that tiles the amyloid surface. Finally, amino acid sequences of the peptide binders are computed to provide a sequence with high geometric and physicochemical complementarity to the target amyloid. This method was applied to a conformational strain of -synuclein fibrils, resulting in a peptide with high specificity for the target relative to other amyloids formed by -synuclein, tau, or A{beta}40. This designed peptide also markedly slowed the formation of -synuclein amyloids. Overall, this method offers a new tool for examining conformational strains of amyloid proteins.
]]></description>
<dc:creator>Wallace, H.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Tan, S. K.</dc:creator>
<dc:creator>Pan, H.</dc:creator>
<dc:creator>Yang, R.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Jo, H.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:creator>Polizzi, N.</dc:creator>
<dc:creator>DeGrado, W.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.14.567090</dc:identifier>
<dc:title><![CDATA[De novo Design of Peptides that Bind Specific Conformers of α-Synuclein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.15.567273v1?rss=1">
<title>
<![CDATA[
Single phage proteins sequester TIR- and cGAS-generated signaling molecules 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.15.567273v1?rss=1</link>
<description><![CDATA[
Prokaryotic anti-phage immune systems use TIR (toll/interleukin-1 receptor) and cGAS (cyclic GMP-AMP synthase) enzymes to produce 1-3/1-2 glycocyclic ADPR (gcADPR) and cyclid di-/tri-nucleotides (CDNs and CTNs) signaling molecules that limit phage replication, respectively 1-3. However, how phages neutralize these common systems is largely unknown. Here, we show that Thoeris anti-defense proteins Tad1 4 and Tad2 5 both have anti-CBASS activity by simultaneously sequestering CBASS cyclic oligonucleotides. Strikingly, apart from binding Thoeris signals 1-3 and 1-2 gcADPR, Tad1 also binds numerous CBASS CDNs/CTNs with high affinity, inhibiting CBASS systems using these molecules in vivo and in vitro. The hexameric Tad1 has six binding sites for CDNs or gcADPR, which are independent from two high affinity binding sites for CTNs. Tad2 also sequesters various CDNs in addition to gcADPR molecules, inhibiting CBASS systems using these CDNs. However, the binding pockets for CDNs and gcADPR are different in Tad2, whereby a tetramer can bind two CDNs and two gcADPR molecules simultaneously. Taken together, Tad1 and Tad2 are both two-pronged inhibitors that, alongside anti-CBASS protein 2, establish a paradigm of phage proteins that flexibly sequester a remarkable breadth of cyclic nucleotides involved in TIR- and cGAS-based anti-phage immunity.
]]></description>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Xiao, Y.</dc:creator>
<dc:creator>Xiong, W.</dc:creator>
<dc:creator>Fedorova, I.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Huiting, E.</dc:creator>
<dc:creator>Ren, J.</dc:creator>
<dc:creator>Gao, Z.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Feng, Y.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.15.567273</dc:identifier>
<dc:title><![CDATA[Single phage proteins sequester TIR- and cGAS-generated signaling molecules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.16.567459v1?rss=1">
<title>
<![CDATA[
Hierarchical annotation of eQTLs enables identification of genes with cell-type divergent regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.16.567459v1?rss=1</link>
<description><![CDATA[
While context-type-specific regulation of genes is largely determined by cis-regulatory regions, attempts to identify cell-type specific eQTLs are complicated by the nested nature of cell types. We present a network-based model for hierarchical annotation of bulk-derived eQTLs to levels of a cell type tree using single cell chromatin accessibility data and no clustering of cells into discrete cell types. Using our model, we annotated bulk-derived eQTLs from the developing brain with high specificity to levels of a cell-type hierarchy. The increased annotation power provided by the hierarchical model allowed for sensitive detection of genes with multiple distinct non-coding elements regulating their expression in different cell types, which we validated in single-cell multiome data and reporter assays. Overall, we find that incorporating the hierarchical organization of cell types provides a powerful way to account for the relationships between cell types in complex tissues.
]]></description>
<dc:creator>Przytycki, P. F.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.16.567459</dc:identifier>
<dc:title><![CDATA[Hierarchical annotation of eQTLs enables identification of genes with cell-type divergent regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.16.566292v1?rss=1">
<title>
<![CDATA[
Networks extracted from nonlinear fMRI connectivity exhibit unique spatial variation and enhanced sensitivity to differences between individuals with schizophrenia and controls 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.16.566292v1?rss=1</link>
<description><![CDATA[
Functional magnetic resonance imaging (fMRI) studies often estimate brain intrinsic connectivity networks (ICNs) from temporal relationships between hemodynamic signals using approaches such as independent component analysis (ICA). While ICNs are thought to represent functional sources that play important roles in various psychological phenomena, current approaches have been tailored to identify ICNs that mainly reflect linear statistical relationships. However, the elements comprising neural systems often exhibit remarkably complex nonlinear interactions that may be involved in cognitive operations and altered in psychiatric conditions such as schizophrenia. Consequently, there is a need to develop methods capable of effectively capturing ICNs from measures that are sensitive to nonlinear relationships. Here, we advance a novel approach to estimate ICNs from explicitly nonlinear whole-brain functional connectivity (ENL-wFC) by transforming resting-state fMRI (rsfMRI) data into the connectivity domain, allowing us to capture unique information from distance correlation patterns that would be missed by linear whole-brain functional connectivity (LIN-wFC) analysis.

Our findings provide evidence that ICNs commonly extracted from linear (LIN) relationships are also reflected in explicitly nonlinear (ENL) connectivity patterns. ENL ICN estimates exhibit higher reliability and stability, highlighting our approachs ability to effectively quantify ICNs from rsfMRI data. Additionally, we observed a consistent spatial gradient pattern between LIN and ENL ICNs with higher ENL weight in core ICN regions, suggesting that ICN function may be subserved by nonlinear processes concentrated within network centers. We also found that a uniquely identified ENL ICN distinguished individuals with schizophrenia from healthy controls while a uniquely identified LIN ICN did not, emphasizing the valuable complementary information that can be gained by incorporating measures that are sensitive to nonlinearity in future analyses. Moreover, the ENL estimates of ICNs associated with auditory, linguistic, sensorimotor, and self-referential processes exhibit heightened sensitivity towards differentiating between individuals with schizophrenia and controls compared to LIN counterparts, demonstrating the translational value of our approach and of the ENL estimates of ICNs that are frequently reported as disrupted in schizophrenia. In summary, our findings underscore the tremendous potential of connectivity domain ICA and nonlinear information in resolving complex brain phenomena and revolutionizing the landscape of clinical FC analysis.
]]></description>
<dc:creator>Kinsey, S.</dc:creator>
<dc:creator>Kazimierczak, K.</dc:creator>
<dc:creator>Camazon, P. A.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Adali, T.</dc:creator>
<dc:creator>Kochunov, P.</dc:creator>
<dc:creator>Adhikari, B.</dc:creator>
<dc:creator>Ford, J.</dc:creator>
<dc:creator>van Erp, T. G. M.</dc:creator>
<dc:creator>Dhamala, M.</dc:creator>
<dc:creator>Calhoun, V. D.</dc:creator>
<dc:creator>Iraji, A.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.16.566292</dc:identifier>
<dc:title><![CDATA[Networks extracted from nonlinear fMRI connectivity exhibit unique spatial variation and enhanced sensitivity to differences between individuals with schizophrenia and controls]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.16.567230v1?rss=1">
<title>
<![CDATA[
Engineered odorant receptors illuminate structural principles of odor discrimination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.16.567230v1?rss=1</link>
<description><![CDATA[
A central challenge in olfaction is understanding how the olfactory system detects and distinguishes odorants with diverse physicochemical properties and molecular configurations. Vertebrate animals perceive odors via G protein-coupled odorant receptors (ORs). In humans, [~]400 ORs enable the sense of smell. The OR family is composed of two major classes: Class I ORs are tuned to carboxylic acids while Class II ORs, representing the vast majority of the human repertoire, respond to a wide variety of odorants. How ORs recognize chemically diverse odorants remains poorly understood. A fundamental bottleneck is the inability to visualize odorant binding to ORs. Here, we uncover fundamental molecular properties of odorant-OR interactions by employing engineered ORs crafted using a consensus protein design strategy. Because such consensus ORs (consORs) are derived from the 17 major subfamilies of human ORs, they provide a template for modeling individual native ORs with high sequence and structural homology. The biochemical tractability of consORs enabled four cryoEM structures of distinct consORs with unique ligand recognition properties. The structure of a Class I consOR, consOR51, showed high structural similarity to the native human receptor OR51E2 and yielded a homology model of a related member of the human OR51 family with high predictive power. Structures of three Class II consORs revealed distinct modes of odorant-binding and activation mechanisms between Class I and Class II ORs. Thus, the structures of consORs lay the groundwork for understanding molecular recognition of odorants by the OR superfamily.
]]></description>
<dc:creator>de March, C. A.</dc:creator>
<dc:creator>Ma, N.</dc:creator>
<dc:creator>Billesboelle, C. B.</dc:creator>
<dc:creator>Tewari, J.</dc:creator>
<dc:creator>del Torrent, C. L.</dc:creator>
<dc:creator>van der Velden, W. J.</dc:creator>
<dc:creator>Ojiro, I.</dc:creator>
<dc:creator>Takayama, I.</dc:creator>
<dc:creator>Faust, B.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Vaidehi, N.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:creator>Matsunami, H.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.16.567230</dc:identifier>
<dc:title><![CDATA[Engineered odorant receptors illuminate structural principles of odor discrimination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.17.567549v1?rss=1">
<title>
<![CDATA[
Reversible histone deacetylase activity catalyzes lysine acylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.17.567549v1?rss=1</link>
<description><![CDATA[
Starvation and low carbohydrate diets lead to the accumulation of the ketone body, {beta}-hydroxybutyrate (BHB), whose blood concentrations increase more than 10-fold into the millimolar range. In addition to providing a carbon source, BHB accumulation triggers lysine {beta}-hydroxybutyrylation (Kbhb) of proteins via unknown mechanisms. As with other lysine acylation events, Kbhb marks can be removed by histone deacetylases (HDACs). Here, we report that class I HDACs unexpectedly catalyze protein lysine modification with {beta}-hydroxybutyrate (BHB). Mutational analyses of the HDAC2 active site reveal a shared reliance on key amino acids for classical deacetylation and non-canonical HDAC-catalyzed {beta}-hydroxybutyrylation. Also consistent with reverse HDAC activity, Kbhb formation is driven by mass action and substrate availability. This reverse HDAC activity is not limited to BHB but also extends to multiple short-chain fatty acids. The reversible activity of class I HDACs described here represents a novel mechanism of PTM deposition relevant to metabolically-sensitive proteome modifications.
]]></description>
<dc:creator>Tsusaka, T.</dc:creator>
<dc:creator>Najar, M. A.</dc:creator>
<dc:creator>Schwarz, B.</dc:creator>
<dc:creator>Bohrnsen, E.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Bosio, C. M.</dc:creator>
<dc:creator>Burslem, G. M.</dc:creator>
<dc:creator>Goldberg, E. L.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.17.567549</dc:identifier>
<dc:title><![CDATA[Reversible histone deacetylase activity catalyzes lysine acylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.17.567593v1?rss=1">
<title>
<![CDATA[
Poc1 is a basal body inner junction protein that promotes triplet microtubule integrity and interconnections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.17.567593v1?rss=1</link>
<description><![CDATA[
Basal bodies (BBs) are conserved eukaryotic structures that organize motile and primary cilia. The BB is comprised of nine, cylindrically arranged, triplet microtubules (TMTs) that are connected to each other by inter-TMT linkages which maintain BB structure. During ciliary beating, forces transmitted to the BB must be resisted to prevent BB disassembly. Poc1 is a conserved BB protein important for BBs to resist ciliary forces. To understand how Poc1 confers BB stability, we identified the precise position of Poc1 binding in the Tetrahymena BB and the effect of Poc1 loss on BB structure. Poc1 binds at the TMT inner junctions, stabilizing TMTs directly. From this location, Poc1 also stabilizes inter-TMT linkages throughout the BB, including the cartwheel pinhead and the inner scaffold. Moreover, we identify a molecular response to ciliary forces via a molecular remodeling of the inner scaffold, as determined by differences in Fam161A localization. Thus, while not essential for BB assembly, Poc1 promotes BB interconnections that establish an architecture competent to resist ciliary forces.
]]></description>
<dc:creator>Ruehle, M. D.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Agard, D.</dc:creator>
<dc:creator>Pearson, C. G.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.17.567593</dc:identifier>
<dc:title><![CDATA[Poc1 is a basal body inner junction protein that promotes triplet microtubule integrity and interconnections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.18.567683v1?rss=1">
<title>
<![CDATA[
An engineered glioblastoma model yields novel macrophage-secreted drivers of invasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.18.567683v1?rss=1</link>
<description><![CDATA[
Glioblastomas (GBMs) are highly invasive brain tumors replete with brain- and blood-derived macrophages, collectively known as tumor-associated macrophages (TAMs). Targeting TAMs has been proposed as a therapeutic strategy but has thus far yielded limited clinical success in slowing GBM progression, due in part to an incomplete understanding of TAM function in GBM. Here, by using an engineered hyaluronic acid-based 3D invasion platform, patient-derived GBM cells, and multi-omics analysis of GBM tumor microenvironments, we show that M2-polarized macrophages stimulate GBM stem cell (GSC) mesenchymal transition and invasion. We identify TAM-derived transforming growth factor beta induced (TGF{beta}I/BIGH3) as a pro-tumorigenic factor in the GBM microenvironment. In GBM patients, BIGH3 mRNA expression correlates with poor patient prognosis and is highest in the most aggressive GBM molecular subtype. Inhibiting TAM-derived BIGH3 signaling with a blocking antibody or small molecule inhibitor suppresses GSC invasion. Our work highlights the utility of 3D in vitro tumor microenvironment platforms to investigate TAM-cancer cell crosstalk and offers new insights into TAM function to guide novel TAM-targeting therapies.
]]></description>
<dc:creator>Akins, E. A.</dc:creator>
<dc:creator>Wilkins, D.</dc:creator>
<dc:creator>Aghi, M.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:date>2023-11-18</dc:date>
<dc:identifier>doi:10.1101/2023.11.18.567683</dc:identifier>
<dc:title><![CDATA[An engineered glioblastoma model yields novel macrophage-secreted drivers of invasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.20.567959v1?rss=1">
<title>
<![CDATA[
Hybrid assemblies of microbiome Blastocystis protists reveal evolutionary diversification reflecting host ecology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.20.567959v1?rss=1</link>
<description><![CDATA[
Blastocystis, an obligate host-associated protist, is the most common microbial eukaryote in the human gut and is widely distributed across vertebrate hosts. The evolutionary transition of Blastocystis from its free-living stramenopile ancestors to a radiation of host-associated organisms is poorly understood. To explore this, we cultured and sequenced eight strains representing the significant phylogenetic diversity of the genus using long-read, short-read, and Hi-C DNA sequencing, alongside gene annotation and RNA sequencing. Comparative genomic analyses revealed significant variation in gene content and genome structure across Blastocystis. Notably, three strains from herbivorous tortoises, phylogenetically distant from human subtypes, have markedly larger genomes with longer introns and intergenic regions, and retain canonical stop codons absent in the human-associated strains. Despite these genetic differences, all eight isolates exhibit gene losses linked to the reduced cellular complexity of Blastocystis, including losses of cilia and flagella genes, microtubule motor genes, and signal transduction genes. Isolates from herbivorous tortoises contained higher numbers of plant carbohydrate-metabolizing enzymes, suggesting that like gut bacteria, these protists ferment plant material in the host gut. We find evidence that some of these carbohydrate-metabolizing enzymes were horizontally acquired from bacteria, indicating that horizontal gene transfer is an ongoing process in Blastocystis that has contributed to host-related adaptation. Together, these results highlight substantial genetic and metabolic diversity within the Blastocystis genus, indicating different lineages of Blastocystis have varied ecological roles in the host gut.
]]></description>
<dc:creator>Lind, A.</dc:creator>
<dc:creator>McDonald, N. A.</dc:creator>
<dc:creator>Gerrick, E. R.</dc:creator>
<dc:creator>Bhatt, A. S.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2023-11-20</dc:date>
<dc:identifier>doi:10.1101/2023.11.20.567959</dc:identifier>
<dc:title><![CDATA[Hybrid assemblies of microbiome Blastocystis protists reveal evolutionary diversification reflecting host ecology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.21.568004v1?rss=1">
<title>
<![CDATA[
Insights into Trypanosomiasis Transmission: Age, Infection Rates and Bloodmeal Analysis of Glossina fuscipes fuscipes in N.W. Uganda 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.21.568004v1?rss=1</link>
<description><![CDATA[
BackgroundTsetse flies (Glossina) transmit species of Trypanosoma which cause human African trypanosomiasis (HAT) and animal African trypanosomiasis (AAT). Measuring the infection status of wild-caught tsetse is an important part of operations to control HAT. In north-west Uganda, we conducted field studies over a 15-month period to compare classical, microscope-based, and PCR-based methods of detecting trypanosomes in tsetse. We also quantified the age structure and host preferences of tsetse.

MethodsUsing Pyramidal traps placed along the Kochi River, in Koboko district, 12512 G. fuscipes fuscipes were caught. A subset of females ({not}n= 5051) and males (n= 1221) underwent dissection wherein mouthparts, midguts and salivary glands were screened for trypanosomes. Additionally, the age of the females was estimated using ovarian ageing and the trypanosome-positive status of 1931 females and 438 males was investigated by ITS PCR. Further the bloodmeal sources of 131 tsetse were identified using vertebrate cytochrome b PCR.

ResultsInfection rates estimated were significantly greater (1.9-9.3 times) using the PCR-based method compared to the classical dissection-based method. Positive rates for T. brucei sl, T. congolense and T. vivax were 1.6% (1.32-2.24), 2.4% (1.83-3.11and 2.0% (1.46-2.63), respectively by PCR. The abundance and age structure of tsetse populations were relatively stable and the slight seasonal four-fold variation in abundance appeared to be correlated with rainfall. Analyses of age structure suggests a low natural daily mortality of 1.75% (1.62-1.88). The majority of bloodmeals were identified as cattle (39%, 30.5-47.8) and human (37% of meals, 28.4-45.6).

ConclusionPCR provides a more sensitive and specific method of estimating the infection rates of all pathogenic species of trypanosome circulating in Koboko. The seasonally stable abundance, low mortality rate and high proportion of bloodmeals from humans may explain, in part, why this district has historically been a focus of sleeping sickness.

Author summaryTsetse flies (Glossina) transmit African trypanosomiasis in humans ( sleeping sickness) and livestock ( nagana). Identifying trypanosome species and quantifying infection rates in tsetse is important in understanding and controlling trypanosomiasis. Traditionally this relied on microscopic examination of tsetse, however, this method is unable to reliably identify different species of trypanosome. We compared microscopy and PCR, trypanosome-detection methods, using G. fuscipes fuscipes collected from traps deployed in Koboko district, in North-West Uganda.

Our results show that the PCR-based method was 1.9-9.3 times more sensitive than the classical approach and able to identify different species of Trypanosoma, including mixed infections. In collecting tsetse we also obtained data on the seasonal abundance, age and diet of the tsetse population. Analyses of these results showed that natural mortality was low (1.75%/day) among adult females and only a slight seasonal variation in abundance and age structure occurred. The most important hosts were cattle and humans, providing 39% and 37% of bloodmeals, respectively.

The PCR method provided a sensitive means of identifying trypanosomes to the species level. The longevity and diet of tsetse in Koboko may explain, why this district was a persistent focus of disease prior to the deployment of Tiny Targets to control tsetse.
]]></description>
<dc:creator>Cunningham, L. J.</dc:creator>
<dc:creator>Esterhuizen, J.</dc:creator>
<dc:creator>Hargrove, J.</dc:creator>
<dc:creator>Lehane, M.</dc:creator>
<dc:creator>Lord, J.</dc:creator>
<dc:creator>Lingley, J.</dc:creator>
<dc:creator>Mangwiro, C. T. N.</dc:creator>
<dc:creator>Opiyo, M.</dc:creator>
<dc:creator>Torr, S.</dc:creator>
<dc:date>2023-11-21</dc:date>
<dc:identifier>doi:10.1101/2023.11.21.568004</dc:identifier>
<dc:title><![CDATA[Insights into Trypanosomiasis Transmission: Age, Infection Rates and Bloodmeal Analysis of Glossina fuscipes fuscipes in N.W. Uganda]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.22.568240v1?rss=1">
<title>
<![CDATA[
Rifampicin tolerance and growth fitness among isoniazid-resistant clinical Mycobacterium tuberculosis isolates: an in-vitro longitudinal study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.22.568240v1?rss=1</link>
<description><![CDATA[
Antibiotic tolerance in Mycobacterium tuberculosis leads to less effective bacterial killing, poor treatment responses and resistant emergence. Therefore, we investigated the rifampicin tolerance of M. tuberculosis isolates, with or without pre-existing isoniazid-resistance. We determined the in-vitro rifampicin survival fraction by minimum duration of killing assay in isoniazid susceptible (IS, n=119) and resistant (IR, n=84) M. tuberculosis isolates. Then we correlated the rifampicin tolerance with bacterial growth, rifampicin minimum inhibitory concentrations (MICs) and isoniazid-resistant mutations. The longitudinal IR isolates collected from patients were analyzed for changes in rifampicin tolerance and associated emergence of genetic variants. The median duration of rifampicin exposure reducing the M. tuberculosis surviving fraction by 90% (minimum duration of killing-MDK90) increased from 1.23 (95%CI 1.11; 1.37) and 1.31 (95%CI 1.14; 1.48) to 2.55 (95%CI 2.04; 2.97) and 1.98 (95%CI 1.69; 2.56) days, for IS and IR respectively, during 15 to 60 days of incubation. This indicated the presence of fast and slow growing tolerant sub-populations. A range of 6 log10-fold survival fraction enabled classification of tolerance as low, medium or high and revealed IR association with increased tolerance with faster growth (OR=2.68 for low vs. medium, OR=4.42 for low vs. high, P-trend=0.0003). The high tolerance in IR isolates was specific to those collected during rifampicin treatment in patients and associated with bacterial genetic microvariants. Furthermore, the high rifampicin tolerant IR isolates have survival potential similar to multi-drug resistant isolates. These findings suggest that IR tuberculosis needs to be evaluated for high rifampicin tolerance to improve treatment regimen and prevent the risk of MDR-TB emergence.
]]></description>
<dc:creator>Srinivasan, V.</dc:creator>
<dc:creator>Bao, N. L. H.</dc:creator>
<dc:creator>Vinh, D. N.</dc:creator>
<dc:creator>Thanh Hoang Le, N.</dc:creator>
<dc:creator>Thu, D. D.</dc:creator>
<dc:creator>Quang, N. L.</dc:creator>
<dc:creator>Trieu, L. P. T.</dc:creator>
<dc:creator>Nhung, H. N.</dc:creator>
<dc:creator>Ha, V. T. N.</dc:creator>
<dc:creator>Thai, P. V. K.</dc:creator>
<dc:creator>Ha, D. T. M.</dc:creator>
<dc:creator>Lan, N. H.</dc:creator>
<dc:creator>Caws, M.</dc:creator>
<dc:creator>Thwaites, G. E.</dc:creator>
<dc:creator>Javid, B.</dc:creator>
<dc:creator>Thuong, N. T.</dc:creator>
<dc:date>2023-11-22</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.568240</dc:identifier>
<dc:title><![CDATA[Rifampicin tolerance and growth fitness among isoniazid-resistant clinical Mycobacterium tuberculosis isolates: an in-vitro longitudinal study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.22.568268v1?rss=1">
<title>
<![CDATA[
ChromaFactor: deconvolution of single-molecule chromatin organization with non-negative matrix factorization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.22.568268v1?rss=1</link>
<description><![CDATA[
The investigation of chromatin organization in single cells holds great promise for identifying causal relationships between genome structure and function. However, analysis of single-molecule data is hampered by extreme yet inherent heterogeneity, making it challenging to determine the contributions of individual chromatin fibers to bulk trends. To address this challenge, we propose ChromaFactor, a novel computational approach based on non-negative matrix factorization that deconvolves single-molecule chromatin organization datasets into their most salient primary components. ChromaFactor provides the ability to identify trends accounting for the maximum variance in the dataset while simultaneously describing the contribution of individual molecules to each component. Applying our approach to two single-molecule imaging datasets across different genomic scales, we find that these primary components demonstrate significant correlation with key functional phenotypes, including active transcription, enhancer-promoter distance, and genomic compartment. ChromaFactor offers a robust tool for understanding the complex interplay between chromatin structure and function on individual DNA molecules, pinpointing which subpopulations drive functional changes and fostering new insights into cellular heterogeneity and its implications for bulk genomic phenomena.
]]></description>
<dc:creator>Gunsalus, L. M.</dc:creator>
<dc:creator>Keiser, M. J.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2023-11-22</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.568268</dc:identifier>
<dc:title><![CDATA[ChromaFactor: deconvolution of single-molecule chromatin organization with non-negative matrix factorization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.22.568314v1?rss=1">
<title>
<![CDATA[
Selective activation of intracellular β1AR using a spatially restricted antagonist 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.22.568314v1?rss=1</link>
<description><![CDATA[
G-protein-coupled receptors (GPCRs) regulate several physiological and pathological processes and represent the target of approximately 30% of FDA-approved drugs. GPCR-mediated signaling was thought to occur exclusively at the plasma membrane. However, recent studies have unveiled their presence and function at subcellular membrane compartments. There is a growing interest in studying compartmentalized signaling of GPCRs. This requires development of novel tools to separate GPCRs signaling at the plasma membrane from the ones initiated at intracellular compartments. We took advantage of the structural and pharmacological information available for {beta}1-adrenergic receptor ({beta}1AR), an exemplary GPCR that functions at subcellular compartments, and rationally designed spatially restricted antagonists. We generated a cell impermeable {beta}1AR antagonist by conjugating a suitable pharmacophore to a sulfonate-containing fluorophore. This cell-impermeable antagonist only inhibited {beta}1AR on the plasma membrane. In contrast, a cell permeable {beta}1AR agonist containing a non-sulfonated fluorophore, efficiently inhibited both the plasma membrane and Golgi pools of {beta}1ARs. Furthermore, the cell impermeable antagonist selectively inhibited the phosphorylation of downstream effectors of PKA proximal to the plasma membrane in adult cardiomyocytes while {beta}1AR intracellular pool remained active. Our tools offer promising avenues for investigating compartmentalized {beta}1AR signaling in various context, potentially advancing our understanding of {beta}1AR-mediated cellular responses in health and disease. They also offer a general strategy to study compartmentalized signaling for other GPCRs in various biological systems.
]]></description>
<dc:creator>Liccardo, F.</dc:creator>
<dc:creator>Morstein, J.</dc:creator>
<dc:creator>Lin, T.-Y.</dc:creator>
<dc:creator>Pampel, J.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:creator>Irannejad, R.</dc:creator>
<dc:date>2023-11-22</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.568314</dc:identifier>
<dc:title><![CDATA[Selective activation of intracellular β1AR using a spatially restricted antagonist]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.22.568334v1?rss=1">
<title>
<![CDATA[
Spatiotemporal dynamics of successive activations across the human brain during simple arithmetic processing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.22.568334v1?rss=1</link>
<description><![CDATA[
Previous neuroimaging studies have offered unique insights about the spatial organization of activations and deactivations across the brain, however these were not powered to explore the exact timing of events at the subsecond scale combined with precise anatomical source information at the level of individual brains. As a result, we know little about the order of engagement across different brain regions during a given cognitive task. Using experimental arithmetic tasks as a prototype for human-unique symbolic processing, we recorded directly across 10,076 brain sites in 85 human subjects (52% female) using intracranial electroencephalography (iEEG). Our data revealed a remarkably distributed change of activity in almost half of the sampled sites. Notably, an orderly successive activation of a set of brain regions - anatomically consistent across subjects-was observed in individual brains. Furthermore, the temporal order of activations across these sites was replicable across subjects and trials. Moreover, the degree of functional connectivity between the sites decreased as a function of temporal distance between regions, suggesting that information is partially leaked or transformed along the processing chain. Furthermore, in each activated region, distinct neuronal populations with opposite activity patterns during target and control conditions were juxtaposed in an anatomically orderly manner. Our study complements the prior imaging studies by providing hitherto unknown information about the timing of events in the brain during arithmetic processing. Such findings can be a basis for developing mechanistic computational models of human-specific cognitive symbolic systems.

Significance statementOur study elucidates the spatiotemporal dynamics and anatomical specificity of brain activations across >10,000 sites during arithmetic tasks, as captured by intracranial EEG. We discovered an orderly, successive activation of brain regions, consistent across individuals, and a decrease in functional connectivity as a function of temporal distance between regions. Our findings provide unprecedented insights into the sequence of cognitive processing and regional interactions, offering a novel perspective for enhancing computational models of cognitive symbolic systems.
]]></description>
<dc:creator>Pinheiro-Chagas, P.</dc:creator>
<dc:creator>Sava-Segal, C.</dc:creator>
<dc:creator>Akkol, S.</dc:creator>
<dc:creator>Daitch, A.</dc:creator>
<dc:creator>Parvizi, J.</dc:creator>
<dc:date>2023-11-22</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.568334</dc:identifier>
<dc:title><![CDATA[Spatiotemporal dynamics of successive activations across the human brain during simple arithmetic processing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.22.568361v1?rss=1">
<title>
<![CDATA[
Assembly reactions of SARS-CoV-2 nucleocapsid protein with nucleic acid 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.22.568361v1?rss=1</link>
<description><![CDATA[
The viral genome of SARS-CoV-2 is packaged by the nucleocapsid (N-) protein into ribonucleoprotein particles (RNPs), 38{+/-}10 of which are contained in each virion. Their architecture has remained unclear due to the pleomorphism of RNPs, the high flexibility of N-protein intrinsically disordered regions, and highly multivalent interactions between viral RNA and N-protein binding sites in both N-terminal (NTD) and C-terminal domain (CTD). Here we explore critical interaction motifs of RNPs by applying a combination of biophysical techniques to mutant proteins binding different nucleic acids in an in vitro assay for RNP formation, and by examining mutant proteins in a viral assembly assay. We find that nucleic acid-bound N-protein dimers oligomerize via a recently described protein-protein interface presented by a transient helix in its long disordered linker region between NTD and CTD. The resulting hexameric complexes are stabilized by multi-valent protein-nucleic acid interactions that establish crosslinks between dimeric subunits. Assemblies are stabilized by the dimeric CTD of N-protein offering more than one binding site for stem-loop RNA. Our study suggests a model for RNP assembly where N- protein scaffolding at high density on viral RNA is followed by cooperative multimerization through protein-protein interactions in the disordered linker.
]]></description>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Syed, A. M.</dc:creator>
<dc:creator>Khalid, M. M.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Ciling, A.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Yau, W.-M.</dc:creator>
<dc:creator>Srinivasan, S.</dc:creator>
<dc:creator>Esposito, D.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:creator>Piszczek, G.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Schuck, P.</dc:creator>
<dc:date>2023-11-23</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.568361</dc:identifier>
<dc:title><![CDATA[Assembly reactions of SARS-CoV-2 nucleocapsid protein with nucleic acid]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.22.568384v1?rss=1">
<title>
<![CDATA[
Gaining Biological Insights through Supervised Data Visualization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.22.568384v1?rss=1</link>
<description><![CDATA[
Dimensionality reduction-based data visualization is pivotal in comprehending complex biological data. The most common methods, such as PHATE, t-SNE, and UMAP, are unsupervised and therefore reflect the dominant structure in the data, which may be independent of expert-provided labels. Here we introduce a supervised data visualization method called RF-PHATE, which integrates expert knowledge for further exploration of the data. RF-PHATE leverages random forests to capture intricate featurelabel relationships. Extracting information from the forest, RF-PHATE generates low-dimensional visualizations that highlight relevant data relationships while disregarding extraneous features. This approach scales to large datasets and applies to classification and regression. We illustrate RF-PHATEs prowess through three case studies. In a multiple sclerosis study using longitudinal clinical and imaging data, RF-PHATE unveils a sub-group of patients with non-benign relapsingremitting Multiple Sclerosis, demonstrating its aptitude for time-series data. In the context of Raman spectral data, RF-PHATE effectively showcases the impact of antioxidants on diesel exhaust-exposed lung cells, highlighting its proficiency in noisy environments. Furthermore, RF-PHATE aligns established geometric structures with COVID-19 patient outcomes, enriching interpretability in a hierarchical manner. RF-PHATE bridges expert insights and visualizations, promising knowledge generation. Its adaptability, scalability, and noise tolerance underscore its potential for widespread adoption.
]]></description>
<dc:creator>Rhodes, J. S.</dc:creator>
<dc:creator>Aumon, A.</dc:creator>
<dc:creator>Morin, S.</dc:creator>
<dc:creator>Girard, M.</dc:creator>
<dc:creator>Larochelle, C.</dc:creator>
<dc:creator>Lahav, B.</dc:creator>
<dc:creator>Brunet-Ratnasingham, E.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Cutler, A.</dc:creator>
<dc:creator>Zhou, A.</dc:creator>
<dc:creator>Kaufmann, D. E.</dc:creator>
<dc:creator>Zandee, S.</dc:creator>
<dc:creator>Prat, A.</dc:creator>
<dc:creator>Wolf, G.</dc:creator>
<dc:creator>Moon, K. R.</dc:creator>
<dc:date>2023-11-23</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.568384</dc:identifier>
<dc:title><![CDATA[Gaining Biological Insights through Supervised Data Visualization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.27.568889v1?rss=1">
<title>
<![CDATA[
Towards Generalizability and Robustness in Biological Object Detection in Electron Microscopy Images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.27.568889v1?rss=1</link>
<description><![CDATA[
Machine learning approaches have the potential for meaningful impact in the biomedical field. However, there are often challenges unique to biomedical data that prohibits the adoption of these innovations. For example, limited data, data volatility, and data shifts all compromise model robustness and generalizability. Without proper tuning and data management, deploying machine learning models in the presence of unaccounted for corruptions leads to reduced or misleading performance. This study explores techniques to enhance model generalizability through iterative adjustments. Specifically, we investigate a detection tasks using electron microscopy images and compare models trained with different normalization and augmentation techniques. We found that models trained with Group Normalization or texture data augmentation outperform other normalization techniques and classical data augmentation, enabling them to learn more generalized features. These improvements persist even when models are trained and tested on disjoint datasets acquired through diverse data acquisition protocols. Results hold true for transformerand convolution-based detection architectures. The experiments show an impressive 29% boost in average precision, indicating significant enhancements in the models generalizibality. This underscores the models capacity to effectively adapt to diverse datasets and demonstrates their increased resilience in real-world applications.
]]></description>
<dc:creator>GIANNIOS, K.</dc:creator>
<dc:creator>Chaurasia, A.</dc:creator>
<dc:creator>Bueno, C.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Pagano, L.</dc:creator>
<dc:creator>Lo, T. P.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>DeLaRosa, B.</dc:creator>
<dc:date>2023-11-27</dc:date>
<dc:identifier>doi:10.1101/2023.11.27.568889</dc:identifier>
<dc:title><![CDATA[Towards Generalizability and Robustness in Biological Object Detection in Electron Microscopy Images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.27.567477v1?rss=1">
<title>
<![CDATA[
Cell-Specific Transposable Element Gene Expression Analysis Identifies Associations with Systemic Lupus Erythematosus Phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.27.567477v1?rss=1</link>
<description><![CDATA[
There is an established yet unexplained link between interferon (IFN) and systemic lupus erythematosus (SLE). The expression of sequences derived from transposable elements (TEs) may contribute to production of type I IFNs and generation of autoantibodies. We profiled cell-sorted RNA-seq data (CD4+ T cells, CD14+ monocytes, CD19+ B cells, and NK cells) from PBMCs of 120 SLE patients and quantified TE expression identifying 27,135 TEs. We tested for differential TE expression across 10 SLE phenotypes including autoantibody production and disease activity and discovered 731 differentially expressed (DE) TEs whose effects were mostly cell-specific and phenotype-specific. DE TEs were enriched for specific families and viral genes encoded in TE sequences. Increased expression of DE TEs was associated with genes involved in antiviral activity such as LY6E, ISG15, TRIM22 and pathways such as interferon signaling. These findings suggest that expression of TEs contributes to activation of SLE-related mechanisms in a cell-specific manner, which can impact disease diagnostics and therapeutics.
]]></description>
<dc:creator>Cutts, Z.</dc:creator>
<dc:creator>Patterson, S.</dc:creator>
<dc:creator>Maliskova, L.</dc:creator>
<dc:creator>Taylor, K. E.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Dall'Era, M.</dc:creator>
<dc:creator>Yazdany, J.</dc:creator>
<dc:creator>Criswell, L.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Lanata, C.</dc:creator>
<dc:date>2023-11-28</dc:date>
<dc:identifier>doi:10.1101/2023.11.27.567477</dc:identifier>
<dc:title><![CDATA[Cell-Specific Transposable Element Gene Expression Analysis Identifies Associations with Systemic Lupus Erythematosus Phenotypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.27.568924v1?rss=1">
<title>
<![CDATA[
Reversal of pulmonary veno-occlusive disease phenotypes by inhibition of the integrated stress response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.27.568924v1?rss=1</link>
<description><![CDATA[
Pulmonary veno-occlusive disease (PVOD) is a rare form of pulmonary hypertension arising from EIF2AK4 gene mutations or mitomycin C (MMC) administration. The lack of effective PVOD therapies is compounded by a limited understanding of the mechanisms driving the vascular remodeling in PVOD. We show that the administration of MMC in rats mediates the activation of protein kinase R (PKR) and the integrated stress response (ISR), which lead to the release of the endothelial adhesion molecule VE-Cadherin in the complex with Rad51 to the circulation, disruption of endothelial barrier, and vascular remodeling. Pharmacological inhibition of PKR or ISR attenuates the depletion of VE-Cadherin, elevation of vascular permeability, and vascular remodeling instigated by MMC, suggesting potential clinical intervention for PVOD. Finally, the severity of PVOD phenotypes was increased by a heterozygous BMPR2 mutation that truncates the carboxyl tail of BMPR2, underscoring the role of deregulated BMP signal in the development of PVOD.
]]></description>
<dc:creator>Prabhakar, A.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Wadhwa, M.</dc:creator>
<dc:creator>Ghatpande, P.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Lizama, C. O.</dc:creator>
<dc:creator>Kharbikar, B. N.</dc:creator>
<dc:creator>Graf, S.</dc:creator>
<dc:creator>Treacy, C. M.</dc:creator>
<dc:creator>Morrell, N. W.</dc:creator>
<dc:creator>Graham, B. B.</dc:creator>
<dc:creator>Lagna, G.</dc:creator>
<dc:creator>Hata, A.</dc:creator>
<dc:date>2023-11-28</dc:date>
<dc:identifier>doi:10.1101/2023.11.27.568924</dc:identifier>
<dc:title><![CDATA[Reversal of pulmonary veno-occlusive disease phenotypes by inhibition of the integrated stress response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.27.568955v1?rss=1">
<title>
<![CDATA[
Epiregulon: Inference of single-cell transcription factor activity to dissect mechanisms of lineage plasticity and drug response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.27.568955v1?rss=1</link>
<description><![CDATA[
Transcription factors (TFs) and transcriptional coregulators represent an emerging class of therapeutic targets in oncology. Gene regulatory networks (GRNs) can be used to evaluate pharmacological agents targeting these factors and to identify drivers of disease and drug resistance. However, GRN methods that rely solely on gene expression often fail to account for post-transcriptional modulation of TF function. We present Epiregulon, a method that constructs GRNs from single-cell ATAC-seq and RNA-seq data for accurate prediction of TF activity. This is achieved by considering the co-occurrence of TF expression and chromatin accessibility at TF binding sites in each cell. We leverage ChIP-seq data to extend inference to transcriptional coregulators lacking defined motifs or TF harboring neomorphic mutations. Epiregulon accurately predicted the effects of AR inhibition across various drug modalities including an AR antagonist and an AR degrader, delineated the mechanisms of a SMARCA4 degrader by identifying context-dependent interaction partners and prioritized known and novel drivers of lineage reprogramming and tumorigenesis. By mapping gene regulation across various cellular contexts, Epiregulon can accelerate the discovery of therapeutics targeting transcriptional regulators.
]]></description>
<dc:creator>Wlodarczyk, T.</dc:creator>
<dc:creator>Lun, A.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Menon, S.</dc:creator>
<dc:creator>Toneyan, S.</dc:creator>
<dc:creator>Seidel, K.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Tan, J.</dc:creator>
<dc:creator>Chen, S.-Y.</dc:creator>
<dc:creator>Keyes, T.</dc:creator>
<dc:creator>Chlebowski, A.</dc:creator>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Metcalfe, C.</dc:creator>
<dc:creator>Hafner, M.</dc:creator>
<dc:creator>Siebel, C. W.</dc:creator>
<dc:creator>Corces, M. R.</dc:creator>
<dc:creator>Yauch, R.</dc:creator>
<dc:creator>Xie, S.</dc:creator>
<dc:creator>Yao, X.</dc:creator>
<dc:date>2023-11-28</dc:date>
<dc:identifier>doi:10.1101/2023.11.27.568955</dc:identifier>
<dc:title><![CDATA[Epiregulon: Inference of single-cell transcription factor activity to dissect mechanisms of lineage plasticity and drug response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.29.568895v1?rss=1">
<title>
<![CDATA[
A (Sub)field Guide to Quality Control in Hippocampal Subfield Segmentation on High-resolution T2-weighted MRI 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.29.568895v1?rss=1</link>
<description><![CDATA[
Inquiries into properties of brain structure and function have progressed due to developments in magnetic resonance imaging (MRI). To sustain progress in investigating and quantifying neuroanatomical details in vivo, the reliability and validity of brain measurements are paramount. Quality control (QC) is a set of procedures for mitigating errors and ensuring the validity and reliability of brain measurements. Despite its importance, there is little guidance on best QC practices and reporting procedures. The study of hippocampal subfields in vivo is a critical case for QC because of their small size, inter-dependent boundary definitions, and common artifacts in the MRI data used for subfield measurements. We addressed this gap by surveying the broader scientific community studying hippocampal subfields on their views and approaches to QC. We received responses from 37 investigators spanning 10 countries, covering different career stages, and studying both healthy and pathological development and aging. In this sample, 81% of researchers considered QC to be very important or important, and 19% viewed it as fairly important. Despite this, only 46% of researchers reported on their QC processes in prior publications. In many instances, lack of reporting appeared due to ambiguous guidance on relevant details and guidance for reporting, rather than absence of QC. Here, we provide recommendations for correcting errors to maximize reliability and minimize bias. We also summarize threats to segmentation accuracy, review common QC methods, and make recommendations for best practices and reporting in publications. Implementing the recommended QC practices will collectively improve inferences to the larger population, as well as have implications for clinical practice and public health.
]]></description>
<dc:creator>Canada, K. L.</dc:creator>
<dc:creator>Mazloum-Farzaghi, N.</dc:creator>
<dc:creator>Radman, G.</dc:creator>
<dc:creator>Adams, J. N.</dc:creator>
<dc:creator>Bakker, A.</dc:creator>
<dc:creator>Baumeister, H.</dc:creator>
<dc:creator>Berron, D.</dc:creator>
<dc:creator>Bocchetta, M.</dc:creator>
<dc:creator>Carr, V.</dc:creator>
<dc:creator>Dalton, M. A.</dc:creator>
<dc:creator>de Flores, R.</dc:creator>
<dc:creator>Keresztes, A.</dc:creator>
<dc:creator>LaJoie, R.</dc:creator>
<dc:creator>Mueller, S.</dc:creator>
<dc:creator>Raz, N.</dc:creator>
<dc:creator>Santini, T.</dc:creator>
<dc:creator>Shaw, T.</dc:creator>
<dc:creator>Stark, C. E. L.</dc:creator>
<dc:creator>Tran, T. T.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Wisse, L.</dc:creator>
<dc:creator>Wuestefeld, A.</dc:creator>
<dc:creator>Yushkevich, P. A.</dc:creator>
<dc:creator>Olsen, R.</dc:creator>
<dc:creator>Daugherty, A.</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.11.29.568895</dc:identifier>
<dc:title><![CDATA[A (Sub)field Guide to Quality Control in Hippocampal Subfield Segmentation on High-resolution T2-weighted MRI]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.29.569312v1?rss=1">
<title>
<![CDATA[
Paradoxical mTORC1-Dependent microRNA-mediated Translation Repression in the Nucleus Accumbens of Mice Consuming Alcohol Attenuates Glycolysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.29.569312v1?rss=1</link>
<description><![CDATA[
mTORC1 promotes protein translation, learning and memory, and neuroadaptations that underlie alcohol use and abuse. We report that activation of mTORC1 in the nucleus accumbens (NAc) of mice consuming alcohol promotes the translation of microRNA (miR) machinery components and the upregulation of microRNAs (miRs) expression including miR-34a-5p. In parallel, we detected a paradoxical mTORC1-dependent repression of translation of transcripts including Aldolase A, an essential glycolytic enzyme. We found that miR-34a-5p in the NAc targets Aldolase A for translation repression and promotes alcohol intake. Our data further suggest that glycolysis is inhibited in the NAc manifesting in an mTORC1-dependent attenuation of L-lactate, the end product of glycolysis. Finally, we show that systemic administration of L-lactate attenuates mouse excessive alcohol intake. Our data suggest that alcohol promotes paradoxical actions of mTORC1 on translation and glycolysis which in turn drive excessive alcohol use.
]]></description>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>Laguesse, S.</dc:creator>
<dc:creator>Phamluong, K.</dc:creator>
<dc:creator>Salvi, A.</dc:creator>
<dc:creator>Hoisington, Z. W.</dc:creator>
<dc:creator>Soneja, D.</dc:creator>
<dc:creator>Sei, Y. J.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.11.29.569312</dc:identifier>
<dc:title><![CDATA[Paradoxical mTORC1-Dependent microRNA-mediated Translation Repression in the Nucleus Accumbens of Mice Consuming Alcohol Attenuates Glycolysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.30.569313v1?rss=1">
<title>
<![CDATA[
The deubiquitinase USP9X regulates RIT1 protein abundance and oncogenic phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.30.569313v1?rss=1</link>
<description><![CDATA[
RIT1 is a rare and understudied oncogene in lung cancer. Despite structural similarity to other RAS GTPase proteins such as KRAS, oncogenic RIT1 activity does not appear to be tightly regulated by nucleotide exchange or hydrolysis. Instead, there is a growing understanding that the protein abundance of RIT1 is important for its regulation and function. We previously identified the deubiquitinase USP9X as a RIT1 dependency in RIT1-mutant cells. Here, we demonstrate that both wild-type and mutant forms of RIT1 are substrates of USP9X. Depletion of USP9X leads to decreased RIT1 protein stability and abundance and resensitizes cells to EGFR tyrosine kinase inhibitors. Our work expands upon the current understanding of RIT1 protein regulation and presents USP9X as a key regulator of RIT1-driven oncogenic phenotypes.
]]></description>
<dc:creator>Riley, A. K.</dc:creator>
<dc:creator>Grant, M.</dc:creator>
<dc:creator>Snell, A.</dc:creator>
<dc:creator>Vichas, A.</dc:creator>
<dc:creator>Moorthi, S.</dc:creator>
<dc:creator>Urisman, A.</dc:creator>
<dc:creator>Castel, P.</dc:creator>
<dc:creator>Wan, L.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.11.30.569313</dc:identifier>
<dc:title><![CDATA[The deubiquitinase USP9X regulates RIT1 protein abundance and oncogenic phenotypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.30.569380v1?rss=1">
<title>
<![CDATA[
Time-of-day effects of drugs revealed by high-throughput deep phenotyping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.30.569380v1?rss=1</link>
<description><![CDATA[
The circadian clock, a fundamental biological regulator, governs essential cellular processes in health and disease. Circadian-based therapeutic strategies are increasingly gaining recognition as promising avenues. Aligning drug administration with the circadian rhythm can enhance treatment efficacy and minimize side effects. Yet, uncovering the optimal treatment timings remains challenging, limiting their widespread adoption. In this work, we introduce a novel high-throughput approach integrating live-imaging and data analysis techniques to deep-phenotype cancer cell models, evaluating their circadian rhythms, growth, and drug responses. We devised a streamlined process for profiling drug sensitivities across different times of the day, identifying optimal treatment windows and responsive cell types and drug combinations. Finally, we implement multiple computational tools to uncover cellular and genetic factors shaping time-of-day drug sensitivity. Our versatile approach is adaptable to various biological models, facilitating its broad application and relevance. Ultimately, this research leverages circadian rhythms to optimize anti-cancer drug treatments, promising improved outcomes and transformative treatment strategies.
]]></description>
<dc:creator>Ector, C.</dc:creator>
<dc:creator>Schmal, C.</dc:creator>
<dc:creator>Didier, J.</dc:creator>
<dc:creator>De Landtsheer, S.</dc:creator>
<dc:creator>Finger, A.-M.</dc:creator>
<dc:creator>Muller-Marquardt, F.</dc:creator>
<dc:creator>Schulte, J.</dc:creator>
<dc:creator>Sauter, T.</dc:creator>
<dc:creator>Keilholz, U.</dc:creator>
<dc:creator>Herzel, H.</dc:creator>
<dc:creator>Kramer, A.</dc:creator>
<dc:creator>Granada, A. E.</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.11.30.569380</dc:identifier>
<dc:title><![CDATA[Time-of-day effects of drugs revealed by high-throughput deep phenotyping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.30.569492v1?rss=1">
<title>
<![CDATA[
Negative feedback control of hunger circuits by the taste of food 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.30.569492v1?rss=1</link>
<description><![CDATA[
The rewarding taste of food is critical for motivating animals to eat, but whether taste has a parallel function in promoting meal termination is not well understood. Here we show that hunger-promoting AgRP neurons are rapidly inhibited during each bout of ingestion by a signal linked to the taste of food. Blocking these transient dips in activity via closed-loop optogenetic stimulation increases food intake by selectively delaying the onset of satiety. We show that upstream leptin receptor-expressing neurons in the dorsomedial hypothalamus (DMHLepR) are tuned to respond to sweet or fatty tastes and exhibit time-locked activation during feeding that is the mirror image of downstream AgRP cells. These findings reveal an unexpected role for taste in the negative feedback control of ingestion. They also reveal a mechanism by which AgRP neurons, which are the primary cells that drive hunger, are able to influence the moment-by-moment dynamics of food consumption.
]]></description>
<dc:creator>Aitken, T. J.</dc:creator>
<dc:creator>Ly, T.</dc:creator>
<dc:creator>Shehata, S.</dc:creator>
<dc:creator>Sivakumar, N.</dc:creator>
<dc:creator>La Santa Medina, N.</dc:creator>
<dc:creator>Gray, L. A.</dc:creator>
<dc:creator>Dundar, N.</dc:creator>
<dc:creator>Barnes, C.</dc:creator>
<dc:creator>Knight, Z. A.</dc:creator>
<dc:date>2023-12-03</dc:date>
<dc:identifier>doi:10.1101/2023.11.30.569492</dc:identifier>
<dc:title><![CDATA[Negative feedback control of hunger circuits by the taste of food]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.03.569805v1?rss=1">
<title>
<![CDATA[
A foundational atlas of autism protein interactions reveals molecular convergence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.03.569805v1?rss=1</link>
<description><![CDATA[
Translating high-confidence (hc) autism spectrum disorder (ASD) genes into viable treatment targets remains elusive. We constructed a foundational protein-protein interaction (PPI) network in HEK293T cells involving 100 hcASD risk genes, revealing over 1,800 PPIs (87% novel). Interactors, expressed in the human brain and enriched for ASD but not schizophrenia genetic risk, converged on protein complexes involved in neurogenesis, tubulin biology, transcriptional regulation, and chromatin modification. A PPI map of 54 patient-derived missense variants identified differential physical interactions, and we leveraged AlphaFold-Multimer predictions to prioritize direct PPIs and specific variants for interrogation in Xenopus tropicalis and human forebrain organoids. A mutation in the transcription factor FOXP1 led to reconfiguration of DNA binding sites and altered development of deep cortical layer neurons in forebrain organoids. This work offers new insights into molecular mechanisms underlying ASD and describes a powerful platform to develop and test therapeutic strategies for many genetically-defined conditions.
]]></description>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Vartak, R.</dc:creator>
<dc:creator>Zaltsman, Y.</dc:creator>
<dc:creator>Naing, Z. Z. C.</dc:creator>
<dc:creator>Hennick, K. M.</dc:creator>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Bashir, A.</dc:creator>
<dc:creator>Eckhardt, M.</dc:creator>
<dc:creator>Bouhaddou, M.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Sun, N.</dc:creator>
<dc:creator>Lasser, M.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Guiley, K. Z.</dc:creator>
<dc:creator>Chan, U.</dc:creator>
<dc:creator>Kaye, J. A.</dc:creator>
<dc:creator>Khare, P.</dc:creator>
<dc:creator>Drake, S.</dc:creator>
<dc:creator>Drury, V.</dc:creator>
<dc:creator>Burke, D. F.</dc:creator>
<dc:creator>Gonzalez, S.</dc:creator>
<dc:creator>Alkhairy, S.</dc:creator>
<dc:creator>Morris, M.</dc:creator>
<dc:creator>Baum, T.</dc:creator>
<dc:creator>Krasnoff, R.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Pham, P.</dc:creator>
<dc:creator>Arbalaez, J.</dc:creator>
<dc:creator>Pratt, D.</dc:creator>
<dc:creator>Chag, S.</dc:creator>
<dc:creator>Rolland, T.</dc:creator>
<dc:creator>Bourgeron, T.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:creator>Bandyopadhay, S.</dc:creator>
<dc:creator>Ideker, T.</dc:creator>
<dc:creator>Beltrao, P.</dc:creator>
<dc:creator>Willsey, H. R.</dc:creator>
<dc:creator>Obernier, K.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Huttenhain, R.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Willsey, A. J.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:date>2023-12-03</dc:date>
<dc:identifier>doi:10.1101/2023.12.03.569805</dc:identifier>
<dc:title><![CDATA[A foundational atlas of autism protein interactions reveals molecular convergence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.01.568932v1?rss=1">
<title>
<![CDATA[
Network-level encoding of local neurotransmitters in cortical astrocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.01.568932v1?rss=1</link>
<description><![CDATA[
Summary ParagraphAstrocytes--the most abundant non-neuronal cell type in the mammalian brain--are crucial circuit components that respond to and modulate neuronal activity via calcium (Ca2+) signaling1-8. Astrocyte Ca2+ activity is highly heterogeneous and occurs across multiple spatiotemporal scales: from fast, subcellular activity3,4 to slow, synchronized activity that travels across connected astrocyte networks9-11. Furthermore, astrocyte network activity has been shown to influence a wide range of processes5,8,12. While astrocyte network activity has important implications for neuronal circuit function, the inputs that drive astrocyte network dynamics remain unclear. Here we used ex vivo and in vivo two-photon Ca2+ imaging of astrocytes while mimicking neuronal neurotransmitter inputs at multiple spatiotemporal scales. We find that brief, subcellular inputs of GABA and glutamate lead to widespread, long-lasting astrocyte Ca2+ responses beyond an individual stimulated cell. Further, we find that a key subset of Ca2+ activity--propagative events--differentiates astrocyte network responses to these two major neurotransmitters, and gates responses to future inputs. Together, our results demonstrate that local, transient neurotransmitter inputs are encoded by broad cortical astrocyte networks over the course of minutes, contributing to accumulating evidence across multiple model organisms that significant astrocyte-neuron communication occurs across slow, network-level spatiotemporal scales13-15. We anticipate that this study will be a starting point for future studies investigating the link between specific astrocyte Ca2+ activity and specific astrocyte functional outputs, which could build a consistent framework for astrocytic modulation of neuronal activity.
]]></description>
<dc:creator>Cahill, M. K.</dc:creator>
<dc:creator>Collard, M.</dc:creator>
<dc:creator>Tse, V.</dc:creator>
<dc:creator>Reitman, M. E.</dc:creator>
<dc:creator>Etchenique, R.</dc:creator>
<dc:creator>Kirst, C.</dc:creator>
<dc:creator>Poskanzer, K. E.</dc:creator>
<dc:date>2023-12-04</dc:date>
<dc:identifier>doi:10.1101/2023.12.01.568932</dc:identifier>
<dc:title><![CDATA[Network-level encoding of local neurotransmitters in cortical astrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.01.569654v1?rss=1">
<title>
<![CDATA[
Atrophy-driven functional network collapse in neurodegenerative disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.01.569654v1?rss=1</link>
<description><![CDATA[
Cognitive and behavioral deficits in Alzheimers disease (AD) and frontotemporal dementia (FTD) result from brain atrophy and altered functional connectivity. However, it is unclear how atrophy relates to functional connectivity disruptions across dementia subtypes and stages. We addressed this question using structural and functional MRI from 221 patients with AD (n=82), behavioral variant FTD (n=41), corticobasal syndrome (n=27), nonfluent (n=34) and semantic (n=37) variant primary progressive aphasia, and 100 cognitively normal individuals. Using partial least squares regression, we identified three principal structure-function components. The first component showed overall atrophy correlating with primary cortical hypo-connectivity and subcortical/association cortical hyper-connectivity. Components two and three linked focal syndrome-specific atrophy to peri-lesional hypo-connectivity and distal hyper-connectivity. Structural and functional component scores predicted global and domain-specific cognitive deficits. Anatomically, functional connectivity changes reflected alterations in specific brain activity gradients. Eigenmode analysis identified temporal phase and amplitude collapse as an explanation for atrophy-driven functional connectivity changes.
]]></description>
<dc:creator>Brown, J. A.</dc:creator>
<dc:creator>Lee, A. J.</dc:creator>
<dc:creator>Fernhoff, K.</dc:creator>
<dc:creator>Pistone, T.</dc:creator>
<dc:creator>Pasquini, L.</dc:creator>
<dc:creator>Wise, A. B.</dc:creator>
<dc:creator>Staffaroni, A. M.</dc:creator>
<dc:creator>Mandelli, M. L.</dc:creator>
<dc:creator>Lee, S. E.</dc:creator>
<dc:creator>Boxer, A. L.</dc:creator>
<dc:creator>Rankin, K. P.</dc:creator>
<dc:creator>Rabinovici, G. D.</dc:creator>
<dc:creator>Gorno-Tempini, M. L.</dc:creator>
<dc:creator>Rosen, H. J.</dc:creator>
<dc:creator>Kramer, J. H.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:date>2023-12-04</dc:date>
<dc:identifier>doi:10.1101/2023.12.01.569654</dc:identifier>
<dc:title><![CDATA[Atrophy-driven functional network collapse in neurodegenerative disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.01.569676v1?rss=1">
<title>
<![CDATA[
Tumor microenvironmental determinants of high-risk DCIS progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.01.569676v1?rss=1</link>
<description><![CDATA[
ABSTRACT/SUMMARYDuctal carcinoma in situ (DCIS) constitutes an array of morphologically recognized intraductal neoplasms in the mammary ductal tree defined by an increased risk for subsequent invasive carcinomas at or near the site of biopsy detection. However, only 15-45% of untreated DCIS cases progress to invasive cancer, so understanding mechanisms that prevent progression is key to avoid overtreatment and provides a basis for alternative therapies and prevention. This study was designed to characterize the tumor microenvironment and molecular profile of high-risk DCIS that grew to a large size but remained as DCIS. All patients had DCIS lesions >5cm in size with at least one additional high-risk feature: young age (<45 years), high nuclear grade, hormone receptor negativity, HER2 positivity, the presence of comedonecrosis, or a palpable mass. The tumor immune microenvironment was characterized using multiplex immunofluorescence to identify immune cells and their spatial relationships within the ducts and stroma. Gene copy number analysis and whole exome DNA sequencing identified the mutational burden and driver mutations, and quantitative whole-transcriptome/gene expression analyses were performed. There was no association between the percent of the DCIS genome characterized by copy number variants (CNAs) and recurrence events (DCIS or invasive). Mutations, especially missense mutations, in the breast cancer driver genes PIK3CA and TP53 were common in this high-risk DCIS cohort (47% of evaluated lesions). Tumor infiltrating lymphocyte (TIL) density was higher in DCIS lesions with TP53 mutations (p=0.0079) compared to wildtype lesions, but not in lesions with PIK3CA mutations (p=0.44). Immune infiltrates were negatively associated with hormone receptor status and positively associated with HER2 expression. High levels of CD3+CD8-T cells were associated with good outcomes with respect to any subsequent recurrence (DCIS or invasive cancer), whereas high levels of CD3+Foxp3+ Treg cells were associated with poor outcomes. Spatial proximity analyses of immune cells and tumor cells demonstrated that close proximity of T cells with tumor cells was associated with good outcomes with respect to any recurrence as well as invasive recurrences. Interestingly, we found that myoepithelial continuity (distance between myoepithelial cells surrounding the involved ducts) was significantly lower in DCIS lesions compared to normal tissue (p=0.0002) or to atypical ductal hyperplasia (p=0.011). Gene set enrichment analysis identified several immune pathways associated with low myoepithelial continuity and a low myoepithelial continuity score was associated with better outcomes, suggesting that gaps in the myoepithelial layer may allow access/interactions between immune infiltrates and tumor cells. Our study demonstrates the immune microenvironment of DCIS, in particular the spatial proximity of tumor cells and T cells, and myoepithelial continuity are important determinants for progression of disease.
]]></description>
<dc:creator>Glencer, A.</dc:creator>
<dc:creator>Ramalingam, K.</dc:creator>
<dc:creator>Schindler, N.</dc:creator>
<dc:creator>Mori, H.</dc:creator>
<dc:creator>Ghule, P.</dc:creator>
<dc:creator>Lee, K. C.</dc:creator>
<dc:creator>Nachmanson, D.</dc:creator>
<dc:creator>Officer, A.</dc:creator>
<dc:creator>Harismendy, O.</dc:creator>
<dc:creator>Stein, J. L.</dc:creator>
<dc:creator>Stein, G.</dc:creator>
<dc:creator>Weaver, D.</dc:creator>
<dc:creator>Yau, C.</dc:creator>
<dc:creator>Hirst, G. L.</dc:creator>
<dc:creator>Campbell, M. J.</dc:creator>
<dc:creator>Esserman, L. J.</dc:creator>
<dc:creator>Borowsky, A. D.</dc:creator>
<dc:date>2023-12-04</dc:date>
<dc:identifier>doi:10.1101/2023.12.01.569676</dc:identifier>
<dc:title><![CDATA[Tumor microenvironmental determinants of high-risk DCIS progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.03.569760v1?rss=1">
<title>
<![CDATA[
Msx genes delineate a novel molecular map of the developing cerebellar neuroepithelium. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.03.569760v1?rss=1</link>
<description><![CDATA[
In the early cerebellar primordium, there are two progenitor zones, the ventricular zone (VZ) residing atop the IVth ventricle and the rhombic lip (RL) at the lateral edges of the developing cerebellum. These zones give rise to the several cell types that form the GABAergic and glutamatergic populations of the adult cerebellum, respectively. Recently, an understanding of the molecular compartmentation of these zones has emerged. The Msx genes are a family of three transcription factors that are expressed downstream of Bone Morphogenetic Protein (BMP) signaling in these zones. Using fluorescent RNA in situ hybridization, we have characterized the Msx (Msh Homeobox) genes and demonstrated that their spatiotemporal pattern segregates specific regions in the progenitor regions. Msx1 and Msx2 are compartmentalized within the rhombic lip (RL), while Msx3 is localized within the ventricular zone (VZ). The relationship of the Msx genes with an early marker of the glutamatergic lineage, Atoh1, was examined in Atoh1-null mice and it was found that the expression of Msx genes persisted. Importantly, Msx1 and Msx3 expressions expanded in response to the elimination of Atoh1. These results point to early markers of cerebellar progenitor zones and more importantly to an updated view of the molecular parcellation of the RL with respect to the canonical marker of the RL, Atoh1.
]]></description>
<dc:creator>Gupta, I.</dc:creator>
<dc:creator>Yeung, J.</dc:creator>
<dc:creator>Rahimi-Balaei, M.</dc:creator>
<dc:creator>Wu, S.-R.</dc:creator>
<dc:creator>Goldowitz, D.</dc:creator>
<dc:date>2023-12-04</dc:date>
<dc:identifier>doi:10.1101/2023.12.03.569760</dc:identifier>
<dc:title><![CDATA[Msx genes delineate a novel molecular map of the developing cerebellar neuroepithelium.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.03.569779v1?rss=1">
<title>
<![CDATA[
Towards A Wireless Image Sensor for Real-Time Fluorescence Microscopy in Cancer Therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.03.569779v1?rss=1</link>
<description><![CDATA[
We present a mm-sized, ultrasonically powered lensless CMOS image sensor as a progress towards wireless fluorescence microscopy. Access to biological information within the tissue has the potential to provide insights guiding diagnosis and treatment across numerous medical conditions including cancer therapy. This information, in conjunction with current clinical imaging techniques that have limitations in obtaining images continuously and lack wireless compatibility, can improve continual detection of multicell clusters deep within tissue. The proposed platform incorporates a 2.4x4.7 mm2 integrated circuit (IC) fabricated in TSMC 0.18 m, a micro laser diode (LD), a single piezoceramic and off-chip storage capacitors. The IC consists of a 36x40 array of capacitive trans-impedance amplifier-based pixels, wireless power management and communication via ultrasound and a laser driver all controlled by a Finite State Machine. The piezoceramic harvests energy from the acoustic waves at a depth of 2 cm to power up the IC and transfer 11.5 kbits/frame via backscattering. During Charge-Up, the off-chip capacitor stores charge to later supply a high-power 78 mW LD during Imaging. Proof of concept of the imaging front end is shown by imaging distributions of CD8 T-cells, an indicator of the immune response to cancer, ex vivo, in the lymph nodes of a functional immune system (BL6 mice) against colorectal cancer consistent with the results of a fluorescence microscope. The overall system performance is verified by detecting 140 m features on a USAF resolution target with 32 ms exposure time and 389 ms ultrasound backscattering.
]]></description>
<dc:creator>Rabbani, R.</dc:creator>
<dc:creator>Najafiaghdam, H.</dc:creator>
<dc:creator>Roschelle, M. L.</dc:creator>
<dc:creator>Papageorgiou, E. P.</dc:creator>
<dc:creator>Zhao, B. R.</dc:creator>
<dc:creator>Ghanbari, M. M.</dc:creator>
<dc:creator>Muller, R.</dc:creator>
<dc:creator>Stojanovic, V.</dc:creator>
<dc:creator>Anwar, M.</dc:creator>
<dc:date>2023-12-05</dc:date>
<dc:identifier>doi:10.1101/2023.12.03.569779</dc:identifier>
<dc:title><![CDATA[Towards A Wireless Image Sensor for Real-Time Fluorescence Microscopy in Cancer Therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.04.569983v1?rss=1">
<title>
<![CDATA[
Understanding how genetically encoded tags affect phase separation by Heterochromatin Protein HP1α 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.04.569983v1?rss=1</link>
<description><![CDATA[
Liquid-liquid phase separation (LLPS) is driven by weak multi-valent interactions. Such interactions can result in the formation of puncta in cells and droplets in vitro. The heterochromatin protein HP1 forms droplets with chromatin in vitro and is found in puncta in cells. A common approach to visualize the dynamics of HP1 in cells is to genetically encode fluorescent tags on the protein. HP1 modified with tags such as GFP has been shown to localize to dynamic puncta in vivo. However, whether tagged HP1 retains its intrinsic phase separation properties has not been systematically studied. Here, using different C-terminal tags (AID-sfGFP, mEGFP, and UnaG), we assessed how tag size and linker length affected the phase separation ability of HP1 with DNA in vitro. We found that the AID-sfGFP tag (52 kDa) promoted HP1 phase-separation, possibly driven by the highly disordered AID degron. The mEGFP tag (27 kDa) inhibited phase-separation by HP1, whereas an UnaG tag (13 kDa) with a 16 amino acid linker showed minimal perturbation. The UnaG tag can thus be used in cellular studies of HP1 to better correlate in vitro and in vivo studies. To test if cellular crowding overcomes the negative effects of large tags in vivo, we used polyethylene glycol (PEG) to mimic crowding in vitro. We found that addition of 10% PEG8000 or PEG4000 enables phase separation by GFP-tagged HP1 at comparable concentrations as untagged HP1. However, these crowding agents also substantially dampened the differences in phase-separation between wild-type and mutant HP1 proteins. PEG further drove phase-separation of Maltose Binding Protein (MBP), a tag often used to solubilize other proteins. These results suggest that phase-separation of biological macromolecules with PEG should be interpreted with caution as PEG-based crowding agents may change the types of interactions made within the phases.
]]></description>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:date>2023-12-05</dc:date>
<dc:identifier>doi:10.1101/2023.12.04.569983</dc:identifier>
<dc:title><![CDATA[Understanding how genetically encoded tags affect phase separation by Heterochromatin Protein HP1α]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.04.570035v1?rss=1">
<title>
<![CDATA[
ADPKD-Causing Missense Variants in Polycystin-1 Disrupt Cell Surface Localization or Polycystin Channel Function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.04.570035v1?rss=1</link>
<description><![CDATA[
Autosomal dominant polycystic kidney disease (ADPKD) is the leading monogenic cause of kidney failure and affects millions of people worldwide. Despite the prevalence of this monogenic disorder, our limited mechanistic understanding of ADPKD has hindered therapeutic development. Here, we successfully developed bioassays that functionally classify missense variants in polycystin-1 (PC1). Strikingly, ADPKD pathogenic missense variants cluster into two major categories: 1) those that disrupt polycystin cell surface localization or 2) those that attenuate polycystin ion channel activity. We found that polycystin channels with defective surface localization could be rescued with a small molecule. We propose that small-molecule-based strategies to improve polycystin cell surface localization and channel function will be effective therapies for ADPKD patients.
]]></description>
<dc:creator>Ha, K.</dc:creator>
<dc:creator>Loeb, G. B.</dc:creator>
<dc:creator>Park, M.</dc:creator>
<dc:creator>Pinedo, A.</dc:creator>
<dc:creator>Park, C. H.</dc:creator>
<dc:creator>Brandes, N.</dc:creator>
<dc:creator>Ritu, F.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:creator>Delling, M.</dc:creator>
<dc:date>2023-12-05</dc:date>
<dc:identifier>doi:10.1101/2023.12.04.570035</dc:identifier>
<dc:title><![CDATA[ADPKD-Causing Missense Variants in Polycystin-1 Disrupt Cell Surface Localization or Polycystin Channel Function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.05.570285v1?rss=1">
<title>
<![CDATA[
Beta and theta oscillations track effort and previous reward in human basal ganglia and prefrontal cortex during decision making 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.05.570285v1?rss=1</link>
<description><![CDATA[
Choosing whether to exert effort to obtain rewards is fundamental to human motivated behavior. However, the neural dynamics underlying the evaluation of reward and effort in humans is poorly understood. Here, we investigate this with chronic intracranial recordings from prefrontal cortex (PFC) and basal ganglia (BG; subthalamic nuclei and globus pallidus) in people with Parkinsons disease performing a decision-making task with offers that varied in levels of reward and physical effort required. This revealed dissociable neural signatures of reward and effort, with BG beta (12-20 Hz) oscillations tracking subjective effort on a single trial basis and PFC theta (4-7 Hz) signaling previous trial reward. Stimulation of PFC increased overall acceptance of offers in addition to increasing the impact of reward on choices. This work uncovers oscillatory mechanisms that guide fundamental decisions to exert effort for reward across BG and PFC, as well as supporting a causal role of PFC for such choices.
]]></description>
<dc:creator>Hoy, C. W.</dc:creator>
<dc:creator>de Hemptinne, C.</dc:creator>
<dc:creator>Wang, S. S.</dc:creator>
<dc:creator>Harmer, C. J.</dc:creator>
<dc:creator>Apps, M. A. J.</dc:creator>
<dc:creator>Husain, M.</dc:creator>
<dc:creator>Starr, P. A.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:date>2023-12-06</dc:date>
<dc:identifier>doi:10.1101/2023.12.05.570285</dc:identifier>
<dc:title><![CDATA[Beta and theta oscillations track effort and previous reward in human basal ganglia and prefrontal cortex during decision making]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.05.570240v1?rss=1">
<title>
<![CDATA[
Fungi and Bacteria Occupy Distinct Spatial Niches within Carious Dentin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.05.570240v1?rss=1</link>
<description><![CDATA[
The role of bacteria in the etiology of dental caries is long established, while the role of fungi has only recently gained more attention. The microbial invasion of dentin in advanced caries especially merits additional research. We evaluated the fungal and bacterial community composition and spatial distribution within carious dentin. Amplicon 16S rRNA gene sequencing together with quantitative PCR was used to profile bacterial and fungal species in caries-free children (n=43) and 4 stages of caries progression from children with severe early childhood caries (n=32). Additionally, healthy (n=10) and carious (n=10) primary teeth were decalcified, sectioned, and stained with Grocotts methenamine silver, periodic acid Schiff (PAS) and calcofluor white (CW) for fungi. Immunolocalization was also performed using antibodies against fungal {beta}-D-glucan, gram-positive bacterial lipoteichoic acid, Streptococcus mutans, and Candida albicans. We also performed field emission scanning electron microscopy (FESEM) to visualize fungi and bacteria within carious dentinal tubules. Bacterial communities observed included a high abundance of S. mutans and the Veillonella parvula group, as expected. There was a higher ratio of fungi to bacteria in dentin-involved lesions compared to less severe lesions with frequent preponderance of C. albicans, C. dubliniensis, and in one case C. tropicalis. Grocotts silver, PAS, CW and immunohistochemistry (IHC) demonstrated the presence of fungi within carious dentinal tubules. Multiplex IHC revealed that fungi and gram-positive bacteria primarily occupied separate dentinal tubules, with rare instances of colocalization. Similar findings were observed with multiplex immunofluorescence using anti-S. mutans and anti-C. albicans antibodies. Electron microscopy showed monomorphic bacterial and fungal biofilms within distinct dentin tubules. We demonstrate a previously unrecognized phenomenon in which fungi and bacteria occupy distinct spatial niches within carious dentin and seldom co-colonize. The potential significance of this phenomenon in caries progression warrants further exploration.
]]></description>
<dc:creator>Sulyanto, R. M.</dc:creator>
<dc:creator>Beall, C. J.</dc:creator>
<dc:creator>Ha, K.</dc:creator>
<dc:creator>Montesano, J.</dc:creator>
<dc:creator>Juang, J.</dc:creator>
<dc:creator>Dickson, J. R.</dc:creator>
<dc:creator>Hashmi, S. B.</dc:creator>
<dc:creator>Bradbury, S.</dc:creator>
<dc:creator>Leys, E. J.</dc:creator>
<dc:creator>Edgerton, M.</dc:creator>
<dc:creator>Ho, S. P.</dc:creator>
<dc:creator>Griffen, A. L.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.05.570240</dc:identifier>
<dc:title><![CDATA[Fungi and Bacteria Occupy Distinct Spatial Niches within Carious Dentin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.06.570164v1?rss=1">
<title>
<![CDATA[
Modeling an ultra-rare epilepsy variant in wildtype mice with in utero prime editing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.06.570164v1?rss=1</link>
<description><![CDATA[
Generating animal models that mirror a patients seizures within clinically-useful timeframes is an important step toward advancing precision medicine for genetic epilepsies. Here we report a somatic cell genome editing approach that rapidly incorporated a patients genomic variant into mice, which developed seizures recapitulating elements of the patients pathology. This approach offers a versatile in vivo platform for clinical, preclinical, and basic research applications, including tailoring pharmacotherapy, assessing variants of uncertain significance, and screening compounds to develop drugs for rare epilepsies. As proof-of-principle, we modeled an epilepsy patient with an ultra-rare variant of the NMDA receptor subunit GRIN2A using prime editing in utero directly in the developing brain of wild-type mice. This methodology achieved high-fidelity genome editing in vivo sufficient to induce frequent spontaneous seizures without necessitating germline modification or extensive breeding. Leveraging the speed and versatility of this approach, we propose a generalizable workflow to generate bedside-to-bench animal models of individual patients within weeks. This advance holds promise for providing a cost-effective, expedient in vivo testing platform that reduces barriers to access for precision medicine, and accelerates drug development for rare and neglected neurological conditions.
]]></description>
<dc:creator>Robertson, C. D.</dc:creator>
<dc:creator>Davis, P.</dc:creator>
<dc:creator>Richardson, R. R.</dc:creator>
<dc:creator>Iffland, P. H.</dc:creator>
<dc:creator>Vieira, D. C. O.</dc:creator>
<dc:creator>Steyert, M.</dc:creator>
<dc:creator>McKeon, P. N.</dc:creator>
<dc:creator>Romanowski, A. J.</dc:creator>
<dc:creator>Crutcher, G.</dc:creator>
<dc:creator>Jasarevic, E.</dc:creator>
<dc:creator>Wolff, S. B. E.</dc:creator>
<dc:creator>Mathur, B. N.</dc:creator>
<dc:creator>Crino, P. B.</dc:creator>
<dc:creator>Bale, T. L.</dc:creator>
<dc:creator>Dick, I. E.</dc:creator>
<dc:creator>Poulopoulos, A.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.06.570164</dc:identifier>
<dc:title><![CDATA[Modeling an ultra-rare epilepsy variant in wildtype mice with in utero prime editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.07.569680v1?rss=1">
<title>
<![CDATA[
MLL3/MLL4 enzymatic activity shapes DNA replication timing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.07.569680v1?rss=1</link>
<description><![CDATA[
Mammalian genomes are replicated in a precise order during S phase, which is cell-type-specific1-3 and correlates with local transcriptional activity2,4-8, chromatin modifications9 and chromatin architecture1,10,11,12. However, the causal relationships between these features and the key regulators of DNA replication timing (RT) are largely unknown. Here, machine learning was applied to quantify chromatin features, including epigenetic marks, histone variants and chromatin architectural factors, best predicting local RT under steady-state and RT changes during early embryonic stem (ES) cell differentiation. About one-third of genome exhibited RT changes during the differentiation. Combined, chromatin features predicted steady-state RT and RT changes with high accuracy. Of these features, histone H3 lysine 4 monomethylation (H3K4me1) catalyzed by MLL3/4 (also known as KMT2C/D) emerged as a top predictor. Loss of Mll3/4 (but not Mll3 alone) or their enzymatic activity resulted in erasure of genome-wide RT dynamics during ES cell differentiation. Sites that normally gain H3K4me1 in a MLL3/4-dependent fashion during the transition failed to transition towards earlier RT, often with transcriptional activation unaffected. Further analysis revealed a requirement for MLL3/4 in promoting DNA replication initiation zones through MCM2 recruitment, providing a direct link for its role in regulating RT. Our results uncover MLL3/4-dependent H3K4me1 as a functional regulator of RT and highlight a causal relationship between the epigenome and RT that is largely uncoupled from transcription. These findings uncover a previously unknown role for MLL3/4-dependent chromatin functions which is likely relevant to the numerous diseases associated with MLL3/4 mutations.
]]></description>
<dc:creator>Goekbuget, D.</dc:creator>
<dc:creator>Boileau, R. M.</dc:creator>
<dc:creator>Lenshoek, K.</dc:creator>
<dc:creator>Blelloch, R.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.07.569680</dc:identifier>
<dc:title><![CDATA[MLL3/MLL4 enzymatic activity shapes DNA replication timing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.07.570544v1?rss=1">
<title>
<![CDATA[
Syndrome-informed phenotyping identifies a polygenic background for achondroplasia-like facial variation in the general population 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.07.570544v1?rss=1</link>
<description><![CDATA[
Human craniofacial shape is highly variable yet highly heritable with genetic variants interacting through multiple layers of development. Here, we hypothesize that Mendelian phenotypes represent the extremes of a phenotypic spectrum and, using achondroplasia as an example, we introduce a syndrome-informed phenotyping approach to identify genomic loci associated with achondroplasia-like facial variation in the normal population. We compared three-dimensional facial scans from 43 individuals with achondroplasia and 8246 controls to calculate achondroplasia-like facial scores. Multivariate GWAS of the control scores revealed a polygenic basis for normal facial variation along an achondroplasia-specific shape axis, identifying genes primarily involved in skeletal development. Jointly modeling these genes in two independent control samples showed craniofacial effects approximating the characteristic achondroplasia phenotype. These findings suggest that both complex and Mendelian genetic variation act on the same developmentally determined axes of facial variation, providing new insights into the genetic intersection of complex traits and Mendelian disorders.
]]></description>
<dc:creator>Vanneste, M.</dc:creator>
<dc:creator>Hoskens, H.</dc:creator>
<dc:creator>Goovaerts, S.</dc:creator>
<dc:creator>Matthews, H.</dc:creator>
<dc:creator>Aponte, J. D.</dc:creator>
<dc:creator>Cole, J.</dc:creator>
<dc:creator>Shriver, M. D.</dc:creator>
<dc:creator>Marazita, M. L.</dc:creator>
<dc:creator>Weinberg, S. M.</dc:creator>
<dc:creator>Walsh, S.</dc:creator>
<dc:creator>Richmond, S.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Spritz, R. A.</dc:creator>
<dc:creator>Peeters, H.</dc:creator>
<dc:creator>Hallgrimsson, B.</dc:creator>
<dc:creator>Claes, P.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.07.570544</dc:identifier>
<dc:title><![CDATA[Syndrome-informed phenotyping identifies a polygenic background for achondroplasia-like facial variation in the general population]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.08.569638v1?rss=1">
<title>
<![CDATA[
Somatic mutation as an explanation for epigenetic aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.08.569638v1?rss=1</link>
<description><![CDATA[
DNA methylation marks have recently been used to build models known as "epigenetic clocks" which predict calendar age. As methylation of cytosine promotes C-to-T mutations, we hypothesized that the methylation changes observed with age should reflect the accrual of somatic mutations, and the two should yield analogous aging estimates. In analysis of multimodal data from 9,331 human individuals, we find that CpG mutations indeed coincide with changes in methylation, not only at the mutated site but also with pervasive remodeling of the methylome out to {+/-}10 kilobases. This one-to-many mapping enables mutation-based predictions of age that agree with epigenetic clocks, including which individuals are aging faster or slower than expected. Moreover, genomic loci where mutations accumulate with age also tend to have methylation patterns that are especially predictive of age. These results suggest a close coupling between the accumulation of sporadic somatic mutations and the widespread changes in methylation observed over the course of life.
]]></description>
<dc:creator>Koch, Z.</dc:creator>
<dc:creator>Li, A. Y.</dc:creator>
<dc:creator>Evans, D.</dc:creator>
<dc:creator>Cummings, S.</dc:creator>
<dc:creator>Ideker, T.</dc:creator>
<dc:date>2023-12-09</dc:date>
<dc:identifier>doi:10.1101/2023.12.08.569638</dc:identifier>
<dc:title><![CDATA[Somatic mutation as an explanation for epigenetic aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.10.570990v1?rss=1">
<title>
<![CDATA[
Torques within and outside the human spindle balance twist at anaphase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.10.570990v1?rss=1</link>
<description><![CDATA[
At each cell division, nanometer-scale motors and microtubules give rise to the micron-scale spindle. Many mitotic motors step helically around microtubules in vitro, and most are predicted to twist the spindle in a left-handed direction. However, the human spindle exhibits only slight global twist, raising the question of how these molecular torques are balanced. Here, using lattice light sheet microscopy, we find that anaphase spindles in the epithelial cell line MCF10A have a high baseline twist, and we identify factors that both increase and decrease this twist. The midzone motors KIF4A and MKLP1 are redundantly required for left-handed twist at anaphase, and we show that KIF4A generates left-handed torque in vitro. The actin cytoskeleton also contributes to left-handed twist, but dynein and its cortical recruitment factor LGN counteract it. Together, our work demonstrates that force generators regulate twist in opposite directions from both within and outside the spindle, preventing strong spindle twist during chromosome segregation.
]]></description>
<dc:creator>Neahring, L.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Cho, N. H.</dc:creator>
<dc:creator>Liu, G.</dc:creator>
<dc:creator>Fernandes, J.</dc:creator>
<dc:creator>Rux, C. J.</dc:creator>
<dc:creator>Nakos, K.</dc:creator>
<dc:creator>Subramanian, R.</dc:creator>
<dc:creator>Upadhyayula, S.</dc:creator>
<dc:creator>Yildiz, A.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2023-12-10</dc:date>
<dc:identifier>doi:10.1101/2023.12.10.570990</dc:identifier>
<dc:title><![CDATA[Torques within and outside the human spindle balance twist at anaphase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.11.571147v1?rss=1">
<title>
<![CDATA[
Cholesterol Dysregulation Drives Seed-Dependent Tau Aggregation in Patient Stem Cell-Derived Models of Tauopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.11.571147v1?rss=1</link>
<description><![CDATA[
Tauopathies are a class of neurodegenerative diseases characterized by the progressive misfolding and accumulation of pathological tau protein in focal regions of the brain, leading to insidious neurodegeneration. Abnormalities in cholesterol metabolism and homeostasis have also been implicated in various neurodegenerative diseases. However, the connection between cholesterol dysregulation and tau pathology remains largely unknown. To model and measure the impact of cholesterol dysregulation on tau, we utilized a combination of in vitro and ex vivo tau aggregation assays using an engineered tau biosensor cell line and human induced pluripotent stem cell (iPSC)-derived neuronal cultures from an individual harboring an autosomal dominant P301L tau mutation and from a healthy control. We demonstrate that excess cholesterol esters lead to an increased rate of tau aggregation in vitro and an increase in seed-dependent insoluble tau aggregates detected in the biosensor line. We observed a strong correlation between cholesterol ester concentration and the presence of high-molecular-weight, oligomeric tau species. Importantly, in tauopathy patient iPSC-derived neurons harboring a P301L tau mutation with endogenous forms of misfolded tau, we show that acute dysregulation of cholesterol homeostasis through acute exposure to human plasma-purified cholesterol esters formed by the linkage of fatty acids to the hydroxyl group of cholesterol leads to the rapid accumulation of phosphorylated tau. Conversely, treatment with the same cholesterol esters pool did not lead to subsequent accumulation of phosphorylated tau in control iPSC-derived neurons. Finally, treatment with a heterobifunctional, small-molecule degrader designed to selectively engage and catalyze the ubiquitination and proteasomal degradation of aberrant tau species prevented cholesterol ester-induced aggregation of tau in the biosensor cell line in a Cereblon E3 ligase-dependent manner. Degrader treatment also restored the resiliency of tauopathy patient-derived neurons towards cholesterol ester-induced tau aggregation phenotypes. Taken together, our study supports a key role of cholesterol dysregulation in tau aggregation. Moreover, it provides further pre-clinical validation of the therapeutic strategy of targeted protein degradation with heterobifunctional tau degraders for blocking tau seeding.
]]></description>
<dc:creator>Lam, M.</dc:creator>
<dc:creator>Kuo, S.-Y.</dc:creator>
<dc:creator>Reis, S.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Silva, M. C.</dc:creator>
<dc:creator>Haggarty, S. J.</dc:creator>
<dc:date>2023-12-12</dc:date>
<dc:identifier>doi:10.1101/2023.12.11.571147</dc:identifier>
<dc:title><![CDATA[Cholesterol Dysregulation Drives Seed-Dependent Tau Aggregation in Patient Stem Cell-Derived Models of Tauopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.11.571166v1?rss=1">
<title>
<![CDATA[
IGHMBP2 deletion suppresses translation and activates the integrated stress response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.11.571166v1?rss=1</link>
<description><![CDATA[
IGHMBP2 is a non-essential, superfamily 1 DNA/RNA helicase that is mutated in patients with rare neuromuscular diseases SMARD1 and CMT2S. IGHMBP2 is implicated in translational and transcriptional regulation via biochemical association with ribosomal proteins, pre-rRNA processing factors, and tRNA-related species. To uncover the cellular consequences of perturbing IGHMBP2, we generated full and partial IGHMBP2 deletion K562 cell lines. Using polysome profiling and a nascent protein synthesis assay, we found that IGHMBP2 deletion modestly reduces global translation. We performed Ribo-seq and RNA-seq and identified diverse gene expression changes due to IGHMBP2 deletion, including ATF4 upregulation. With recent studies showing the ISR can contribute to tRNA metabolism-linked neuropathies, we asked whether perturbing IGHMBP2 promotes ISR activation. We generated ATF4 reporter cell lines and found IGHMBP2 knockout cells demonstrate basal, chronic ISR activation. Our work expands upon the impact of IGHMBP2 in translation and elucidates molecular mechanisms that may link mutant IGHMBP2 to severe clinical phenotypes.
]]></description>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Desai, H.</dc:creator>
<dc:creator>Liboy-Lugo, J.</dc:creator>
<dc:creator>Gu, S.</dc:creator>
<dc:creator>Jowhar, Z.</dc:creator>
<dc:creator>Xu, A.</dc:creator>
<dc:creator>Floor, S. N.</dc:creator>
<dc:date>2023-12-12</dc:date>
<dc:identifier>doi:10.1101/2023.12.11.571166</dc:identifier>
<dc:title><![CDATA[IGHMBP2 deletion suppresses translation and activates the integrated stress response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.12.571198v1?rss=1">
<title>
<![CDATA[
Glioma Cells Secrete Collagen VI to Facilitate Invasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.12.571198v1?rss=1</link>
<description><![CDATA[
While glioblastoma (GBM) progression is associated with extensive extracellular matrix (ECM) secretion, the causal contributions of ECM secretion to invasion remain unclear. Here we investigate these contributions by combining engineered materials, proteomics, analysis of patient data, and a model of bevacizumab-resistant GBM. We find that GBM cells cultured in engineered 3D hyaluronic acid hydrogels secrete ECM prior to invasion, particularly in the absence of exogenous ECM ligands. Proteomic measurements reveal extensive secretion of collagen VI, and collagen VI-associated transcripts are correspondingly enriched in microvascular proliferation regions of human GBMs. We further show that bevacizumab-resistant GBM cells deposit more collagen VI than their responsive counterparts, which is associated with marked cell-ECM stiffening. COL6A3 deletion in GBM cells reduces invasion, {beta}-catenin signaling, and expression of mesenchymal markers, and these effects are amplified in hypoxia. Our studies strongly implicate GBM cell-derived collagen VI in microenvironmental remodeling to facilitate invasion.
]]></description>
<dc:creator>Cha, J.</dc:creator>
<dc:creator>Ding, E. A.</dc:creator>
<dc:creator>Carvalho, E. M.</dc:creator>
<dc:creator>Fowler, A.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:date>2023-12-12</dc:date>
<dc:identifier>doi:10.1101/2023.12.12.571198</dc:identifier>
<dc:title><![CDATA[Glioma Cells Secrete Collagen VI to Facilitate Invasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.15.571823v1?rss=1">
<title>
<![CDATA[
Structure-conditioned masked language models for protein sequence design generalize beyond the native sequence space 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.15.571823v1?rss=1</link>
<description><![CDATA[
Machine learning has revolutionized computational protein design, enabling significant progress in protein backbone generation and sequence design. Here, we introduce Frame2seq, a structure-conditioned masked language model for protein sequence design. Frame2seq generates sequences in a single pass, achieves 49.1% sequence recovery on the CATH 4.2 test dataset, and accurately estimates the error in its own predictions, outperforming the autoregressive ProteinMPNN model with over six times faster inference. To probe the ability of Frame2seq to generate novel designs beyond the native-like sequence space it was trained on, we experimentally test 26 Frame2seq designs for de novo backbones with low identity to the starting sequences. We show that Frame2seq successfully designs soluble (22/26), monomeric, folded, and stable proteins (17/26), including a design with 0% sequence identity to native. The speed and accuracy of Frame2seq will accelerate exploration of novel sequence space across diverse design tasks, including challenging applications such as multi-objective optimization.
]]></description>
<dc:creator>Akpinaroglu, D.</dc:creator>
<dc:creator>Seki, K.</dc:creator>
<dc:creator>Guo, A.</dc:creator>
<dc:creator>Zhu, E.</dc:creator>
<dc:creator>Kelly, M. J.</dc:creator>
<dc:creator>Kortemme, T.</dc:creator>
<dc:date>2023-12-15</dc:date>
<dc:identifier>doi:10.1101/2023.12.15.571823</dc:identifier>
<dc:title><![CDATA[Structure-conditioned masked language models for protein sequence design generalize beyond the native sequence space]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.27.573420v1?rss=1">
<title>
<![CDATA[
The Primary Cilium and its Hedgehog Signaling in Nociceptors Contribute to Inflammatory and Neuropathic Pain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.27.573420v1?rss=1</link>
<description><![CDATA[
The primary cilium, a 1-3 m long hair-like structure protruding from the surface of almost all cells in the vertebrate body, is critical for neuronal development and also functions in the adult. As the migratory neural crest settles into dorsal root ganglia (DRG) sensory neurons elaborate a single primary cilium at their soma that is maintained into adult stages. While it is not known if primary cilia are expressed in nociceptors, or their potential function in the mature DRG neuron, recent studies have shown a role for Hedgehog, whose signaling demonstrates a dependence on primary cilia, in nociceptor sensitization. Here we report the expression of primary cilia in rat and mouse nociceptors, where they modulate mechanical nociceptive threshold, and contribute to inflammatory and neuropathic pain. When siRNA targeting Ift88, a primary cilium-specific intra-flagellar transport (IFT) protein required for ciliary integrity, was administered by intrathecal injection, in the rat, it resulted in loss of Ift88 mRNA in DRG, and primary cilia in neuronal cell bodies, which was associated with an increase in mechanical nociceptive threshold, and abrogation of hyperalgesia induced by the pronociceptive inflammatory mediator, prostaglandin E2, and painful peripheral neuropathy induced by a neurotoxic chemotherapy drug, paclitaxel. To provide further support for the role of the primary cilium in nociceptor function we also administered siRNA for another IFT protein, Ift52. Ift52 siRNA results in loss of Ift52 in DRG and abrogates paclitaxel-induced painful peripheral neuropathy. Attenuation of Hedgehog-induced hyperalgesia by Ift88 knockdown supports a role for the primary cilium in the hyperalgesia induced by Hedgehog, and attenuation of paclitaxel chemotherapy-induced neuropathy (CIPN) by cyclopamine, which attenuates Hedgehog signaling, suggests a role of Hedgehog in CIPN. Our findings support a role of nociceptor primary cilia in the control of mechanical nociceptive threshold and in inflammatory and neuropathic pain, the latter, at least in part, Hedgehog dependent.
]]></description>
<dc:creator>Fitzsimons, L. A.</dc:creator>
<dc:creator>Staurengo-Ferrari, L.</dc:creator>
<dc:creator>Bogen, O.</dc:creator>
<dc:creator>Araldi, D.</dc:creator>
<dc:creator>Bonet, I. J. M.</dc:creator>
<dc:creator>Jordan, E. E.</dc:creator>
<dc:creator>Levine, J. D.</dc:creator>
<dc:creator>Tucker, K. L.</dc:creator>
<dc:date>2023-12-27</dc:date>
<dc:identifier>doi:10.1101/2023.12.27.573420</dc:identifier>
<dc:title><![CDATA[The Primary Cilium and its Hedgehog Signaling in Nociceptors Contribute to Inflammatory and Neuropathic Pain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.27.573448v1?rss=1">
<title>
<![CDATA[
Structure-based discovery of positive allosteric modulators for the calcium sensing receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.27.573448v1?rss=1</link>
<description><![CDATA[
Drugs acting as positive allosteric modulators (PAMs) to enhance the activation of the calcium sensing receptor (CaSR) and to suppress parathyroid hormone (PTH) secretion can treat hyperparathyroidism but suffer from side effects including hypocalcemia and arrhythmias. Seeking new CaSR modulators, we docked libraries of 2.7 million and 1.2 billion molecules against transforming pockets in the active-state receptor dimer structure. Consistent with simulations suggesting that docking improves with library size, billion-molecule docking found new PAMs with a hit rate that was 2.7-fold higher than the million-molecule library and with hits up to 37-fold more potent. Structure-based optimization of ligands from both campaigns led to nanomolar leads, one of which was advanced to animal testing. This PAM displays 100-fold the potency of the standard of care, cinacalcet, in ex vivo organ assays, and reduces serum PTH levels in mice by up to 80% without the hypocalcemia typical of CaSR drugs. Cryo-EM structures with the new PAMs show that they induce residue rearrangements in the binding pockets and promote CaSR dimer conformations that are closer to the G-protein coupled state compared to established drugs. These findings highlight the promise of large library docking for therapeutic leads, especially when combined with experimental structure determination and mechanism.

One sentence summaryStructure-based virtual screening uncovers novel CaSR allosteric modulators with enhanced efficacy and less side effects.
]]></description>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>Wu, C.-G.</dc:creator>
<dc:creator>Tu, C.-L.</dc:creator>
<dc:creator>Glenn, I.</dc:creator>
<dc:creator>Meyerowitz, J.</dc:creator>
<dc:creator>Kaplan, A. L.</dc:creator>
<dc:creator>Lyu, J.</dc:creator>
<dc:creator>Cheng, Z.</dc:creator>
<dc:creator>Tarkhanova, O. O.</dc:creator>
<dc:creator>Moroz, Y. S.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Chang, W.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Skiniotis, G.</dc:creator>
<dc:date>2023-12-27</dc:date>
<dc:identifier>doi:10.1101/2023.12.27.573448</dc:identifier>
<dc:title><![CDATA[Structure-based discovery of positive allosteric modulators for the calcium sensing receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.27.573461v1?rss=1">
<title>
<![CDATA[
Mapping cis- and trans-regulatory target genes of human-specific deletions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.27.573461v1?rss=1</link>
<description><![CDATA[
Deletion of functional sequence is predicted to represent a fundamental mechanism of molecular evolution1,2. Comparative genetic studies of primates2,3 have identified thousands of human-specific deletions (hDels), and the cis-regulatory potential of short ([&le;]31 base pairs) hDels has been assessed using reporter assays4. However, how structural variant-sized ([&ge;]50 base pairs) hDels influence molecular and cellular processes in their native genomic contexts remains unexplored. Here, we design genome-scale libraries of single-guide RNAs targeting 7.2 megabases of sequence in 6,358 hDels and present a systematic CRISPR interference (CRISPRi) screening approach to identify hDels that modify cellular proliferation in chimpanzee pluripotent stem cells. By intersecting hDels with chromatin state features and performing single-cell CRISPRi (Perturb-seq) to identify their cis- and trans-regulatory target genes, we discovered 20 hDels controlling gene expression. We highlight two hDels, hDel_2247 and hDel_585, with tissue-specific activity in the brain. Our findings reveal a molecular and cellular role for sequences lost in the human lineage and establish a framework for functionally interrogating human-specific genetic variants.
]]></description>
<dc:creator>Fair, T.</dc:creator>
<dc:creator>Pavlovic, B. J.</dc:creator>
<dc:creator>Schaefer, N. K.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:date>2023-12-28</dc:date>
<dc:identifier>doi:10.1101/2023.12.27.573461</dc:identifier>
<dc:title><![CDATA[Mapping cis- and trans-regulatory target genes of human-specific deletions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.31.573797v1?rss=1">
<title>
<![CDATA[
Conditioned overconsumption is dependent on reinforcer type in lean, but not obese, mice. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.31.573797v1?rss=1</link>
<description><![CDATA[
Associative learning can drive many different types of behaviors, including food consumption. Previous studies have shown that cues paired with food delivery while mice are hungry will lead increased consumption in the presence of those cues at later times. We previously showed that overconsumption can be driven in male mice by contextual cues, using chow pellets. Here we extended our findings by examining other parameters that may influence the outcome of context-conditioned overconsumption training. We found that the task worked equally well in males and females, and that palatable substances such as high-fat diet and Ensure chocolate milkshake supported learning and induced overconsumption. Surprisingly, mice did not overconsume when sucrose was used as the reinforcer during training, suggesting that nutritional content is a critical factor. Interestingly, we also observed that diet-induced obese mice did not learn the task. Overall, we find that context-conditioned overconsumption can be studied in lean males and female mice, and with multiple reinforcer types.
]]></description>
<dc:creator>Lewis-Sanders, D.</dc:creator>
<dc:creator>Bullich, S.</dc:creator>
<dc:creator>Olvera, M. J.</dc:creator>
<dc:creator>Vo, J.</dc:creator>
<dc:creator>Hwang, Y.-S.</dc:creator>
<dc:creator>Stern, S. A.</dc:creator>
<dc:date>2024-01-01</dc:date>
<dc:identifier>doi:10.1101/2023.12.31.573797</dc:identifier>
<dc:title><![CDATA[Conditioned overconsumption is dependent on reinforcer type in lean, but not obese, mice.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.14.575609v1?rss=1">
<title>
<![CDATA[
Blood immunophenotyping identifies distinct kidney histopathology and outcomes in patients with lupus nephritis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.14.575609v1?rss=1</link>
<description><![CDATA[
Lupus nephritis (LN) is a frequent manifestation of systemic lupus erythematosus, and fewer than half of patients achieve complete renal response with standard immunosuppressants. Identifying non-invasive, blood-based pathologic immune alterations associated with renal injury could aid therapeutic decisions. Here, we used mass cytometry immunophenotyping of peripheral blood mononuclear cells in 145 patients with biopsy-proven LN and 40 healthy controls to evaluate the heterogeneity of immune activation in patients with LN and to identify correlates of renal parameters and treatment response. Unbiased analysis identified 3 immunologically distinct groups of patients with LN that were associated with different patterns of histopathology, renal cell infiltrates, urine proteomic profiles, and treatment response at one year. Patients with enriched circulating granzyme B+ T cells at baseline showed more severe disease and increased numbers of activated CD8 T cells in the kidney, yet they had the highest likelihood of treatment response. A second group characterized primarily by a high type I interferon signature had a lower likelihood of response to therapy, while a third group appeared immunologically inactive by immunophenotyping at enrollment but with chronic renal injuries. Main immune profiles could be distilled down to 5 simple cytometric parameters that recapitulate several of the associations, highlighting the potential for blood immune profiling to translate to clinically useful non-invasive metrics to assess immune-mediated disease in LN.
]]></description>
<dc:creator>Horisberger, A.</dc:creator>
<dc:creator>Griffith, A.</dc:creator>
<dc:creator>Keegan, J.</dc:creator>
<dc:creator>Arazi, A.</dc:creator>
<dc:creator>Pulford, J.</dc:creator>
<dc:creator>Murzin, E.</dc:creator>
<dc:creator>Howard, K.</dc:creator>
<dc:creator>Hancock, B.</dc:creator>
<dc:creator>Fava, A.</dc:creator>
<dc:creator>Sasaki, T.</dc:creator>
<dc:creator>Ghosh, T.</dc:creator>
<dc:creator>Inamo, J.</dc:creator>
<dc:creator>Beuschel, R.</dc:creator>
<dc:creator>Cao, Y.</dc:creator>
<dc:creator>Preisinger, K.</dc:creator>
<dc:creator>Gutierrez-Arcelus, M.</dc:creator>
<dc:creator>Eisenhaure, T. M.</dc:creator>
<dc:creator>Guthridge, J.</dc:creator>
<dc:creator>Hoover, P. J.</dc:creator>
<dc:creator>Dall'Era, M.</dc:creator>
<dc:creator>Wofsy, D.</dc:creator>
<dc:creator>Kamen, D. L.</dc:creator>
<dc:creator>Kalunian, K. C.</dc:creator>
<dc:creator>Furie, R.</dc:creator>
<dc:creator>Belmont, M.</dc:creator>
<dc:creator>Izmirly, P.</dc:creator>
<dc:creator>Clancy, R.</dc:creator>
<dc:creator>Hildeman, D.</dc:creator>
<dc:creator>Woodle, E. S.</dc:creator>
<dc:creator>Apruzzese, W.</dc:creator>
<dc:creator>McMahon, M. A.</dc:creator>
<dc:creator>Grossman, J.</dc:creator>
<dc:creator>Barnas, J. L.</dc:creator>
<dc:creator>Payan-Schober, F.</dc:creator>
<dc:creator>Ishimori, M.</dc:creator>
<dc:creator>Weisman, M.</dc:creator>
<dc:creator>Kretzler, M.</dc:creator>
<dc:creator>Berthier, C. C.</dc:creator>
<dc:creator>Hodgin, J. B.</dc:creator>
<dc:creator>Demeke, D. S.</dc:creator>
<dc:creator>Putterman, C.</dc:creator>
<dc:creator>Accelerating Medicines Pa</dc:creator>
<dc:date>2024-01-16</dc:date>
<dc:identifier>doi:10.1101/2024.01.14.575609</dc:identifier>
<dc:title><![CDATA[Blood immunophenotyping identifies distinct kidney histopathology and outcomes in patients with lupus nephritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.15.575686v1?rss=1">
<title>
<![CDATA[
Host brain environmental influences on transplanted medial ganglionic eminence progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.15.575686v1?rss=1</link>
<description><![CDATA[
Interneuron progenitor transplantation can ameliorate disease symptoms in a variety of neurological disorders. This strategy is based on transplantation of embryonic medial ganglionic eminence (MGE) progenitors. Elucidating host brain environment influences on interneuron progenitors as they integrate is critical to optimizing this strategy across different disease states. Here, we systematically evaluated age and brain region influences on survival, migration and differentiation of transplant-derived cells. We find that early postnatal MGE transplantation yields superior survival and more extensive migratory capabilities compared to juvenile or adult. MGE progenitors migrate more widely in cortex compared to hippocampus. Maturation to interneuron subtypes is regulated by age and brain region. MGE progenitors transplanted into dentate gyrus sub-region of early postnatal hippocampus can differentiate into astrocytes. Our results suggest that host brain environment critically regulates survival, spatial distribution and maturation of MGE-derived interneurons following transplantation. These findings inform and enable optimal conditions for interneuron transplant therapies.
]]></description>
<dc:creator>Paterno, R.</dc:creator>
<dc:creator>Vu, T.</dc:creator>
<dc:creator>Hsieh, C.</dc:creator>
<dc:creator>Baraban, S. C.</dc:creator>
<dc:date>2024-01-16</dc:date>
<dc:identifier>doi:10.1101/2024.01.15.575686</dc:identifier>
<dc:title><![CDATA[Host brain environmental influences on transplanted medial ganglionic eminence progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.15.575766v1?rss=1">
<title>
<![CDATA[
Opioid-driven disruption of the septal complex reveals a role for neurotensin- expressing neurons in withdrawal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.15.575766v1?rss=1</link>
<description><![CDATA[
Because opioid withdrawal is an intensely aversive experience, persons with opioid use disorder (OUD) often relapse to avoid it. The lateral septum (LS) is a forebrain structure that is important in aversion processing, and previous studies have linked the lateral septum (LS) to substance use disorders. It is unclear, however, which precise LS cell types might contribute to the maladaptive state of withdrawal. To address this, we used single-nucleus RNA-sequencing to interrogate cell type specific gene expression changes induced by chronic morphine and withdrawal. We discovered that morphine globally disrupted the transcriptional profile of LS cell types, but Neurotensin-expressing neurons (Nts; LS-Nts neurons) were selectively activated by naloxone. Using two-photon calcium imaging and ex vivo electrophysiology, we next demonstrate that LS-Nts neurons receive enhanced glutamatergic drive in morphine-dependent mice and remain hyperactivated during opioid withdrawal. Finally, we showed that activating and silencing LS-Nts neurons during opioid withdrawal regulates pain coping behaviors and sociability. Together, these results suggest that LS-Nts neurons are a key neural substrate involved in opioid withdrawal and establish the LS as a crucial regulator of adaptive behaviors, specifically pertaining to OUD.
]]></description>
<dc:creator>Simon, R.</dc:creator>
<dc:creator>Fleming, W.</dc:creator>
<dc:creator>Senthilkumar, P.</dc:creator>
<dc:creator>Briones, B.</dc:creator>
<dc:creator>Ishii, K.</dc:creator>
<dc:creator>Hjort, M.</dc:creator>
<dc:creator>Martin, M.</dc:creator>
<dc:creator>Hashikawa, K.</dc:creator>
<dc:creator>Sanders, A.</dc:creator>
<dc:creator>Golden, S.</dc:creator>
<dc:creator>Stuber, G.</dc:creator>
<dc:date>2024-01-16</dc:date>
<dc:identifier>doi:10.1101/2024.01.15.575766</dc:identifier>
<dc:title><![CDATA[Opioid-driven disruption of the septal complex reveals a role for neurotensin- expressing neurons in withdrawal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.15.575794v1?rss=1">
<title>
<![CDATA[
A call for a unified and multimodal definition of cellular identity in the enteric nervous system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.15.575794v1?rss=1</link>
<description><![CDATA[
The enteric nervous system (ENS) is a tantalizing frontier in neuroscience. With the recent emergence of single cell transcriptomic technologies, this rare and poorly understood tissue has begun to be better characterized in recent years. A precise functional mapping of enteric neuron diversity is critical for understanding ENS biology and enteric neuropathies. Nonetheless, this pursuit has faced considerable technical challenges. By leveraging different methods to compare available primary mouse and human ENS datasets, we underscore the urgent need for careful identity annotation, achieved through the harmonization and advancements of wet lab and computational techniques. We took different approaches including differential gene expression, module scoring, co-expression and correlation analysis, unbiased biological function hierarchical clustering, data integration and label transfer to compare and contrast functional annotations of several independently reported ENS datasets. These analyses highlight substantial discrepancies stemming from an overreliance on transcriptomics data without adequate validation in tissues. To achieve a comprehensive understanding of enteric neuron identity and their functional context, it is imperative to expand tissue sources and incorporate innovative technologies such as multiplexed imaging, electrophysiology, spatial transcriptomics, as well as comprehensive profiling of epigenome, proteome, and metabolome. Harnessing human pluripotent stem cell (hPSC) models provides unique opportunities for delineating lineage trees of the human ENS, and offers unparalleled advantages, including their scalability and compatibility with genetic manipulation and unbiased screens. We encourage a paradigm shift in our comprehension of cellular complexity and function in the ENS by calling for large-scale collaborative efforts and research investments.
]]></description>
<dc:creator>Majd, H.</dc:creator>
<dc:creator>Cesiulis, A.</dc:creator>
<dc:creator>Samuel, R. M.</dc:creator>
<dc:creator>Richter, M. N.</dc:creator>
<dc:creator>Elder, N.</dc:creator>
<dc:creator>Guyer, R. A.</dc:creator>
<dc:creator>Hao, M.</dc:creator>
<dc:creator>Stamp, L.</dc:creator>
<dc:creator>Goldstein, A.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:date>2024-01-16</dc:date>
<dc:identifier>doi:10.1101/2024.01.15.575794</dc:identifier>
<dc:title><![CDATA[A call for a unified and multimodal definition of cellular identity in the enteric nervous system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.16.575956v1?rss=1">
<title>
<![CDATA[
Molecular and cellular dynamics of the developing human neocortex at single-cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.16.575956v1?rss=1</link>
<description><![CDATA[
The development of the human neocortex is a highly dynamic process and involves complex cellular trajectories controlled by cell-type-specific gene regulation1. Here, we collected paired single-nucleus chromatin accessibility and transcriptome data from 38 human neocortical samples encompassing both the prefrontal cortex and primary visual cortex. These samples span five main developmental stages, ranging from the first trimester to adolescence. In parallel, we performed spatial transcriptomic analysis on a subset of the samples to illustrate spatial organization and intercellular communication. This atlas enables us to catalog cell type-, age-, and area-specific gene regulatory networks underlying neural differentiation. Moreover, combining single-cell profiling, progenitor purification, and lineage-tracing experiments, we have untangled the complex lineage relationships among progenitor subtypes during the transition from neurogenesis to gliogenesis in the human neocortex. We identified a tripotential intermediate progenitor subtype, termed Tri-IPC, responsible for the local production of GABAergic neurons, oligodendrocyte precursor cells, and astrocytes. Remarkably, most glioblastoma cells resemble Tri-IPCs at the transcriptomic level, suggesting that cancer cells hijack developmental processes to enhance growth and heterogeneity. Furthermore, by integrating our atlas data with large-scale GWAS data, we created a disease-risk map highlighting enriched ASD risk in second-trimester intratelencephalic projection neurons. Our study sheds light on the gene regulatory landscape and cellular dynamics of the developing human neocortex.
]]></description>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Moriano, J. A.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Mukhtar, T.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Cebrian-Silla, A.</dc:creator>
<dc:creator>Bi, Q.</dc:creator>
<dc:creator>Augustin, J. J.</dc:creator>
<dc:creator>de Oliveira, L. G.</dc:creator>
<dc:creator>Song, M.</dc:creator>
<dc:creator>Ge, X.</dc:creator>
<dc:creator>Zuo, G.</dc:creator>
<dc:creator>Paredes, M. F.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:creator>Duan, X.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Kriegstein, A. R.</dc:creator>
<dc:date>2024-01-16</dc:date>
<dc:identifier>doi:10.1101/2024.01.16.575956</dc:identifier>
<dc:title><![CDATA[Molecular and cellular dynamics of the developing human neocortex at single-cell resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.18.576113v1?rss=1">
<title>
<![CDATA[
Targeted DNA Sequencing of Cutaneous Melanoma Identifies Prognostic and Predictive Alterations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.18.576113v1?rss=1</link>
<description><![CDATA[
ImportanceCutaneous melanoma (CM) can be molecularly classified into four groups: BRAF mutant, NRAS mutant, NF1 loss, and triple wild type (TWT) tumors lacking any of these three alterations. In the era of immune checkpoint inhibition (ICI) and targeted molecular therapy, the clinical significance of these groups remains unclear. Here, we integrate targeted DNA sequencing with comprehensive clinical follow-up in CM patients.

ObjectiveTo explore how molecular features and tumor mutational burden (TMB) impact outcomes in patients with cutaneous melanoma.

DesignThis was a retrospective cohort study that assessed clinical and molecular features from patients with localized or metastatic CM who underwent targeted next-generation sequencing as part of routine clinical care.

SettingTertiary referral, comprehensive NCI cancer center from 2013 - 2023.

ParticipantsA total of 254 patients with CM who had a CLIA certified targeted sequencing assay performed on their tumor tissue were included

ExposureA CLIA certified targeted sequencing assay was performed as standard of care on 254 patients with CM treated at a single institution.

Main OutcomeNRAS mutation correlated with significantly worse overall survival compared to other TCGA driver groups. Elevated TMB correlated with improved progression-free survival on combination checkpoint inhibition (anti-PD1 plus anti-CTLA4).

ResultsOf 254 patients with cutaneous melanoma, 77 were BRAF mutant (30.3%), 77 were NRAS mutant (30.3%), 47 were NF1 mutant (18.5%), 33 were TWT (13.0%) and the remaining 20 (7.9%) carried mutations in multiple driver genes (BRAF/NRAS/NF1 co-mutated). The majority of this co-mutation group carried mutations in NF1 (n=19 or 90%) with co-occurring mutations in BRAF or NRAS, often with a weaker oncogenic variant. Consistently, NF1 mutant tumors harbored numerous significantly co-altered genes compared to BRAF or NRAS mutant tumors. The majority of TWT tumors (n=29, 87.9%) harbored a pathogenic mutation within a known Ras/MAPK signaling pathway component. Of the 154 cases with available TMB data, the median TMB was 20 (range 0.7 - 266 mutations/Mb). A total of 14 cases (9.1%) were classified as TMB low (<5 mutations/Mb), 64 of 154 (41.6%) were TMB intermediate (>5 and <20 mutations/Mb), 40 of 154 (26.0%) were TMB high (>20 and <50 mutations/Mb) and 36 of 154 (23.4%) were classified as TMB very high (>50 mutations/Mb). NRAS mutant melanoma demonstrated significantly decreased overall survival on multivariable analysis (HR for death 2.95, 95% CI 1.13 - 7.69, p = 0.027, log rank test) compared with other TCGA molecular subgroups. Other factors correlated with decreased overall survival included age and ECOG score. Of the 116 patients in our cohort with available treatment data, 36 received combination dual ICI with anti-CTLA4 and anti-PD1 inhibition as first line therapy. Elevated TMB was associated with significantly longer progression-free survival following dual agent ICI (HR 0.26, 95% CI 0.07 - 0.90, p =0.033, log rank test).

Conclusions and RelevanceNRAS mutation in CMs correlated with significantly worse overall survival. Elevated TMB was associated with increased progression-free survival for patients treated with combination dual ICI, supporting the potential utility of TMB as a predictive biomarker for ICI response in melanoma.
]]></description>
<dc:creator>Vasudevan, H.</dc:creator>
<dc:creator>Haugh, A. M.</dc:creator>
<dc:creator>Osorio, R. C.</dc:creator>
<dc:creator>Francois, R. A.</dc:creator>
<dc:creator>Tawil, M. E.</dc:creator>
<dc:creator>Tsai, K. K.</dc:creator>
<dc:creator>Tetzlaff, M.</dc:creator>
<dc:creator>Daud, A.</dc:creator>
<dc:date>2024-01-22</dc:date>
<dc:identifier>doi:10.1101/2024.01.18.576113</dc:identifier>
<dc:title><![CDATA[Targeted DNA Sequencing of Cutaneous Melanoma Identifies Prognostic and Predictive Alterations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.18.576133v1?rss=1">
<title>
<![CDATA[
Ultra-efficient, unified discovery from microbial sequencing with SPLASH and precise statistical assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.18.576133v1?rss=1</link>
<description><![CDATA[
Bacteria comprise > 12% of Earths biomass and profoundly impact human and planetary health.1 Many key biological functions of microbes, and functions differentiating strains, are conferred or modified by genome plasticity including mobilization of genetic elements, phage integration, and CRISPR arrays. Characterizing each of these processes is time-consuming and requires custom bioinformatic workflows ill-suited to enable discovery of new sources of genetic diversity or to uncover which elements are active. Further, strain typing of bacterial species and approaches to discriminate sub-populations remain time-consuming and resource intensive. Here, we show that SPLASH, our published approach for reference-free discovery and analysis directly from raw reads, and an improved statistical assembly algorithm, compactors, unify diverse tasks in microbial sequence analysis: discovering new mobile elements and CRISPR arrays missing from any reference, and generating rapid, metadata-free strain typing of diverse bacteria. SPLASH and compactors together constitute a new general discovery tool for biological discovery in the microbial world.
]]></description>
<dc:creator>Henderson, G.</dc:creator>
<dc:creator>Gudys, A.</dc:creator>
<dc:creator>Baharav, T.</dc:creator>
<dc:creator>Sundaramurthy, P.</dc:creator>
<dc:creator>Kokot, M.</dc:creator>
<dc:creator>Wang, P. L.</dc:creator>
<dc:creator>Deorowicz, S.</dc:creator>
<dc:creator>Carey, A.</dc:creator>
<dc:creator>Salzman, J.</dc:creator>
<dc:date>2024-01-22</dc:date>
<dc:identifier>doi:10.1101/2024.01.18.576133</dc:identifier>
<dc:title><![CDATA[Ultra-efficient, unified discovery from microbial sequencing with SPLASH and precise statistical assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.22.576634v1?rss=1">
<title>
<![CDATA[
Maternal n-3 enriched diet reprograms neurovascular transcriptome and blunts inflammation in neonate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.22.576634v1?rss=1</link>
<description><![CDATA[
Infection during perinatal period can adversely affect brain development, predispose infants to ischemic stroke and have lifelong consequences. We previously demonstrated that diet enriched in n-3 polyunsaturated fatty acids (PUFA) transforms brain lipid composition and protects from neonatal stroke. Vasculature is a critical interface between blood and brain providing a barrier to systemic infection. Here we examined whether maternal PUFA-enriched diets exert reprograming of endothelial cell signalling in 9-day old mice after endotoxin (LPS)-induced infection. Transcriptome analysis was performed on brain microvessels from pups born to dams maintained on 3 diets: standard, n-3 or n-6 enriched. N-3 diet enabled higher immune reactivity in brain vasculature, while preventing imbalance of cell cycle regulation and extracellular matrix cascades that accompanied inflammatory response in standard diet. LPS response in blood and brain was blunted in n-3 offspring. Cerebral angioarchitecture analysis revealed modified vessel complexity after LPS. Thus, n-3-enriched maternal diet partially prevents imbalance in homeostatic processes and alters inflammation rather than affects brain vascularization during early life. Importantly, maternal diet may presage offspring neurovascular outcomes later in life.
]]></description>
<dc:creator>Chumak, T.</dc:creator>
<dc:creator>Jullienne, A.</dc:creator>
<dc:creator>Ek, J.</dc:creator>
<dc:creator>Ardalan, M.</dc:creator>
<dc:creator>Svedin, P.</dc:creator>
<dc:creator>Quan, R.</dc:creator>
<dc:creator>Salehi, A.</dc:creator>
<dc:creator>Salari, S.</dc:creator>
<dc:creator>Obenaus, A.</dc:creator>
<dc:creator>Vexler, Z.</dc:creator>
<dc:creator>Mallard, C.</dc:creator>
<dc:date>2024-01-22</dc:date>
<dc:identifier>doi:10.1101/2024.01.22.576634</dc:identifier>
<dc:title><![CDATA[Maternal n-3 enriched diet reprograms neurovascular transcriptome and blunts inflammation in neonate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.23.576971v1?rss=1">
<title>
<![CDATA[
Widespread Peptide Surfactants with Post-translational C-methylations Promote Bacterial Development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.23.576971v1?rss=1</link>
<description><![CDATA[
Bacteria produce a variety of peptides to mediate nutrient acquisition, microbial interactions, and other physiological processes. Of special interest are surface-active peptides that aid in growth and development. Herein, we report the structure and characterization of clavusporins, unusual and hydrophobic ribosomal peptides with multiple C-methylations at unactivated carbon centers, which help drastically reduce the surface tension of water and thereby aid in Streptomyces development. The peptides are synthesized by a previously uncharacterized protein superfamily, termed DUF5825, in conjunction with a vitamin B12-dependent radical S-adenosylmethionine metalloenzyme. The operon encoding clavusporin is wide-spread among actinomycete bacteria, suggesting a prevalent role for clavusporins as morphogens in erecting aerial hyphae and thereby advancing sporulation and proliferation.
]]></description>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Seyedsayamdost, M. R.</dc:creator>
<dc:date>2024-01-24</dc:date>
<dc:identifier>doi:10.1101/2024.01.23.576971</dc:identifier>
<dc:title><![CDATA[Widespread Peptide Surfactants with Post-translational C-methylations Promote Bacterial Development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.24.576921v1?rss=1">
<title>
<![CDATA[
High-resolution Inference of Multiplexed Anti-HIV Gene Editing using Single-Cell Targeted DNA Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.24.576921v1?rss=1</link>
<description><![CDATA[
Gene therapy-based HIV cure strategies typically aim to excise the HIV provirus directly, or target host dependency factors (HDFs) that support viral persistence. Cure approaches will likely require simultaneous co-targeting of multiple sites within the HIV genome to prevent evolution of resistance, and/or co-targeting of multiple HDFs to fully render host cells refractory to HIV infection. Bulk cell-based methods do not enable inference of co-editing within individual viral or target cell genomes, and do not discriminate between monoallelic and biallelic gene disruption. Here, we describe a targeted single-cell DNA sequencing (scDNA-seq) platform characterizing the near full-length HIV genome and 50 established HDF genes, designed to evaluate anti-HIV gene therapy strategies. We implemented the platform to investigate the capacity of multiplexed CRISPR-Cas9 ribonucleoprotein complexes (Cas9-RNPs) to simultaneously 1) inactivate the HIV provirus, and 2) knockout the CCR5 and CXCR4 HDF (entry co-receptor) genes in microglia and primary monocyte-derived macrophages (MDMs). Our scDNA-seq pipeline revealed that antiviral gene editing is rarely observed at multiple loci (or both alleles of a locus) within an individual cell, and editing probabilities across sites are linked. Our results demonstrate that single-cell sequencing is critical to evaluate the true efficacy and therapeutic potential of HIV gene therapy.
]]></description>
<dc:creator>Bouzidi, M.</dc:creator>
<dc:creator>Dossani, Z.</dc:creator>
<dc:creator>Di Benedetto, C.</dc:creator>
<dc:creator>Raymond, K.</dc:creator>
<dc:creator>Desai, S.</dc:creator>
<dc:creator>Chavez, L.</dc:creator>
<dc:creator>Betancur, P.</dc:creator>
<dc:creator>Pillai, S. K.</dc:creator>
<dc:date>2024-01-24</dc:date>
<dc:identifier>doi:10.1101/2024.01.24.576921</dc:identifier>
<dc:title><![CDATA[High-resolution Inference of Multiplexed Anti-HIV Gene Editing using Single-Cell Targeted DNA Sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.25.576941v1?rss=1">
<title>
<![CDATA[
Regional specialization manifests in the reliability of neural population codes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.25.576941v1?rss=1</link>
<description><![CDATA[
The brain has the remarkable ability to learn and guide the performance of complex tasks. Decades of lesion studies suggest that different brain regions perform specialized functions in support of complex behaviors1-3. Yet recent large-scale studies of neural activity reveal similar patterns of activity and encoding distributed widely throughout the brain4-6. How these distributed patterns of activity and encoding are compatible with regional specialization of brain function remains unclear. Two frontal brain regions, the dorsal medial prefrontal cortex (dmPFC) and orbitofrontal cortex (OFC), are a paradigm of this conundrum. In the setting complex behaviors, the dmPFC is necessary for choosing optimal actions2,7,8, whereas the OFC is necessary for waiting for3,9 and learning from2,7,9-12 the outcomes of those actions. Yet both dmPFC and OFC encode both choice- and outcome-related quantities13-20. Here we show that while ensembles of neurons in the dmPFC and OFC of rats encode similar elements of a cognitive task with similar patterns of activity, the two regions differ in when that coding is consistent across trials ("reliable"). In line with the known critical functions of each region, dmPFC activity is more reliable when animals are making choices and less reliable preceding outcomes, whereas OFC activity shows the opposite pattern. Our findings identify the dynamic reliability of neural population codes as a mechanism whereby different brain regions may support distinct cognitive functions despite exhibiting similar patterns of activity and encoding similar quantities.
]]></description>
<dc:creator>Guidera, J. A.</dc:creator>
<dc:creator>Gramling, D. P.</dc:creator>
<dc:creator>Comrie, A. E.</dc:creator>
<dc:creator>Joshi, A.</dc:creator>
<dc:creator>Denovellis, E. L.</dc:creator>
<dc:creator>Lee, K. H.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Thompson, P.</dc:creator>
<dc:creator>Hernandez, J.</dc:creator>
<dc:creator>Yorita, A.</dc:creator>
<dc:creator>Haque, R.</dc:creator>
<dc:creator>Kirst, C.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.576941</dc:identifier>
<dc:title><![CDATA[Regional specialization manifests in the reliability of neural population codes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.25.577267v1?rss=1">
<title>
<![CDATA[
An experimental census of retrons for DNA production and genome editing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.25.577267v1?rss=1</link>
<description><![CDATA[
Retrons are bacterial immune systems that use reverse transcribed DNA as a detector of phage infection. They are also increasingly deployed as a component of biotechnology. For genome editing, for instance, retrons are modified so that the reverse transcribed DNA (RT-DNA) encodes an editing donor. Retrons are commonly found in bacterial genomes; thousands of unique retrons have now been predicted bioinformatically. However, only a small number have been characterized experimentally. Here, we add substantially to the corpus of experimentally studied retrons. We synthesized >100 previously untested retrons to identify the natural sequence of RT-DNA they produce, quantify their RT-DNA production, and test the relative efficacy of editing using retron-derived donors to edit bacterial genomes, phage genomes, and human genomes. We add 62 new empirically determined, natural RT-DNAs, which are not predictable from the retron sequence alone. We report a large diversity in RT-DNA production and editing rates across retrons, finding that top performing editors outperform those used in previous studies, and are drawn from a subset of the retron phylogeny.
]]></description>
<dc:creator>Khan, A. G.</dc:creator>
<dc:creator>Rojas-Montero, M.</dc:creator>
<dc:creator>Gonzalez-Delgado, A.</dc:creator>
<dc:creator>Lopez, S. C.</dc:creator>
<dc:creator>Fang, R. F.</dc:creator>
<dc:creator>Shipman, S. L.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.577267</dc:identifier>
<dc:title><![CDATA[An experimental census of retrons for DNA production and genome editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.25.577274v1?rss=1">
<title>
<![CDATA[
Oscillatory Hypoxia Induced Unfolded Protein Folding Response Gene Expression Predicts Low Survival in Human Breast Cancer Patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.25.577274v1?rss=1</link>
<description><![CDATA[
Hypoxia is one of the key factors in the tumor microenvironment regulating nearly all steps in the metastatic cascade in many cancers, including in breast cancer. The hypoxic regions can however be dynamic with the availability of oxygen fluctuating or oscillating. The canonical response to hypoxia is relayed by transcription factor HIF-1, which is stabilized in hypoxia and acts as the master regulator of a large number of downstream genes. However, HIF-1 transcriptional activity can also fluctuate either due to unstable hypoxia, or by lactate mediated non-canonical degradation of HIF-1. Our understanding of how oscillatory hypoxia or HIF-1 activity specifically influence cancer malignancy is very limited. Here, using MDA-MB-231 cells as a model of triple negative breast cancer characterized by severe hypoxia, we measured the gene expression changes induced specifically by oscillatory hypoxia. We found that oscillatory hypoxia can specifically regulate gene expression differently, and at times opposite to stable hypoxia. Using The Cancer Genome Atlas (TCGA) RNAseq data of human cancer samples, we show that the oscillatory specific gene expression signature in MDA-MB-231 is enriched in most human cancers, and prognosticate low survival in breast cancer patients. In particular, we found that oscillatory hypoxia, unlike stable hypoxia, induces unfolded protein folding response (UPR) in cells resulting in gene expression predicting reduced survival.
]]></description>
<dc:creator>Suhail, Y.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Du, W.</dc:creator>
<dc:creator>Afzal, J.</dc:creator>
<dc:creator>Atiq, A.</dc:creator>
<dc:creator>Kshitiz,</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.577274</dc:identifier>
<dc:title><![CDATA[Oscillatory Hypoxia Induced Unfolded Protein Folding Response Gene Expression Predicts Low Survival in Human Breast Cancer Patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.25.577277v1?rss=1">
<title>
<![CDATA[
CDX2 dose-dependently influences the gene regulatory network underlying human extraembryonic mesoderm development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.25.577277v1?rss=1</link>
<description><![CDATA[
Proper regulation of gene dosage is critical for the development of the early embryo and the extraembryonic tissues that support it. Specifically, loss of Cdx2 in vivo leads to stunted development of the allantois, an extraembryonic mesoderm-derived structure critical for nutrient delivery and waste removal in the early embryo. In this study, we investigate how CDX2 dose-dependently influences the gene regulatory network underlying extraembryonic mesoderm development. We generate an allelic series for CDX2 in human induced pluripotent stem cells (hiPSCs) consisting of WT, heterozygous, and homozygous null CDX2 genotypes, differentiate these cells in a 2D gastruloid model, and subject these cells to multiomic single nucleus RNA and ATAC sequencing. We identify several genes that CDX2 dose-dependently regulate cytoskeletal integrity and adhesiveness in the extraembryonic mesoderm population, including regulators of the VEGF, canonical WNT, and non-canonical WNT signaling pathways. Despite these dose-dependent gene expression patterns, snATAC-seq reveals that heterozygous CDX2 expression is capable of inducing a WT-like chromatin accessibility profile, suggesting accessibility is not sufficient to drive gene expression when the CDX2 dosage is reduced. Finally, because the loss of CDX2 or TBXT phenocopy one another in vivo, we compare differentially expressed genes in our CDX2 knock-out model to those from TBXT knock-out hiPSCs differentiated in an analogous experiment. This comparison identifies several communally misregulated genes that are critical for cytoskeletal integrity and tissue permeability, including ANK3 and ANGPT1. Together, these results clarify how CDX2 dose-dependently regulates gene expression in the extraembryonic mesoderm and suggest these genes may underlie the defects in vascular development and allantoic elongation seen in the absence or reduction of CDX2 in vivo.

Summary StatementUsing 2D human gastruloids, CDX2 is shown to dose-dependently influence genes related to tissue permeability, cell-cell adhesions, and cytoskeletal architecture during extraembryonic mesoderm development.
]]></description>
<dc:creator>Bulger, E. A.</dc:creator>
<dc:creator>McDevitt, T. C.</dc:creator>
<dc:creator>Bruneau, B. G.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.577277</dc:identifier>
<dc:title><![CDATA[CDX2 dose-dependently influences the gene regulatory network underlying human extraembryonic mesoderm development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.25.577293v1?rss=1">
<title>
<![CDATA[
Vagus nerve stimulation (VNS) modulates synaptic plasticity in the rat infralimbic cortex via Trk-B receptor activation to reduce drug-seeking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.25.577293v1?rss=1</link>
<description><![CDATA[
Drugs of abuse cause changes in the prefrontal cortex (PFC) and associated regions that impair inhibitory control over drug-seeking. Breaking the contingencies between drug-associated cues and the delivery of the reward during extinction learning reduces relapse. Vagus nerve stimulation (VNS) has previously been shown to enhance extinction learning and reduce drug-seeking. Here we determined the effects of VNS-mediated release of brain-derived neurotrophic factor (BDNF) on extinction and cue-induced reinstatement in rats trained to self-administer cocaine. Pairing 10 days of extinction training with VNS facilitated extinction and reduced drug-seeking behavior during reinstatement. Rats that received a single extinction session with VNS showed elevated BDNF levels in the medial PFC as determined via an enzyme-linked immunosorbent assay (ELISA). Systemic blockade of Tropomyosin receptor kinase B (TrkB) receptors during extinction, via the TrkB antagonist ANA-12, decreased the effects of VNS on extinction and reinstatement. Whole-cell recordings in brain slices showed that cocaine self-administration induced alterations in the ratio of AMPA and NMDA receptor-mediated currents in layer 5 pyramidal neurons of the infralimbic cortex (IL). Pairing extinction with VNS reversed cocaine-induced changes in glutamatergic transmission by enhancing AMPAR currents, and this effect was blocked by ANA-12. Our study suggests that VNS consolidates extinction of drug-seeking behavior by reversing drug-induced changes in synaptic AMPA receptors in the IL, and this effect is abolished by blocking TrkB receptors during extinction, highlighting a potential mechanism for the therapeutic effects of VNS in addiction.

Significance StatementExtinction training can reverse maladaptive neuroplasticity induced by drugs of abuse, but adjunct treatments are sought that can facilitate the process and consolidate the newly formed memories. Pairing extinction training with vagus nerve stimulation (VNS) facilitates extinction and reduces drug-seeking behavior during reinstatement. Here, we show that rats receiving a single extinction session with VNS exhibit elevated brain-derived neurotrophic factor (BDNF) levels in the medial prefrontal cortex (mPFC). We also demonstrate that VNS consolidates the extinction of drug-seeking behavior by reversing cocaine-induced changes in synaptic AMPA receptors in the infralimbic cortex (IL) of the mPFC. This effect is blocked by the TrkB antagonist ANA-12, emphasizing the role of BDNF and TrkB receptors in the therapeutic effects of VNS in addiction.
]]></description>
<dc:creator>Driskill, C. M.</dc:creator>
<dc:creator>Childs, J. E.</dc:creator>
<dc:creator>Phensy, A. J.</dc:creator>
<dc:creator>Rodriguez, S. R.</dc:creator>
<dc:creator>O'Brien, J. T.</dc:creator>
<dc:creator>Lindquist, K. L.</dc:creator>
<dc:creator>Naderi, A. O.</dc:creator>
<dc:creator>Bordieanu, B.</dc:creator>
<dc:creator>McGinty, J. F.</dc:creator>
<dc:creator>Kroener, S.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.577293</dc:identifier>
<dc:title><![CDATA[Vagus nerve stimulation (VNS) modulates synaptic plasticity in the rat infralimbic cortex via Trk-B receptor activation to reduce drug-seeking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.25.577295v1?rss=1">
<title>
<![CDATA[
Spyglass: a data analysis framework for reproducible and shareable neuroscience research 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.25.577295v1?rss=1</link>
<description><![CDATA[
Scientific progress depends on reliable and reproducible results. Progress can be accelerated when data are shared and re-analyzed to address new questions. Current approaches to storing and analyzing neural data involve bespoke formats and software that make replication and reuse of data difficult. To address these challenges, we created Spyglass, an open-source data management and analysis framework written in Python. Spyglass provides reproducible pipelines for common neuroscience analyses and sharing of raw data, intermediate analyses, and final results within and across labs. Spyglass uses the Neurodata Without Borders (NWB) standard and includes pipelines for spectral filtering, spike sorting, pose tracking, and neural decoding. Spyglass can be extended to apply existing and newly developed pipelines to datasets from multiple sources. We demonstrate these features in the context of a cross-laboratory replication by applying advanced state space decoding algorithms to publicly available data.

New users can try out Spyglass on a Jupyter Hub hosted by HHMI and 2i2c: https://spyglass.hhmi.2i2c.cloud/.
]]></description>
<dc:creator>Lee, K. H.</dc:creator>
<dc:creator>Denovellis, E. L.</dc:creator>
<dc:creator>Ly, R.</dc:creator>
<dc:creator>Magland, J.</dc:creator>
<dc:creator>Soules, J.</dc:creator>
<dc:creator>Comrie, A. E.</dc:creator>
<dc:creator>Gramling, D. P.</dc:creator>
<dc:creator>Guidera, J. A.</dc:creator>
<dc:creator>Nevers, R.</dc:creator>
<dc:creator>Adenekan, P.</dc:creator>
<dc:creator>Brozdowski, C.</dc:creator>
<dc:creator>Bray, S.</dc:creator>
<dc:creator>Monroe, E.</dc:creator>
<dc:creator>Bak, J. H.</dc:creator>
<dc:creator>Coulter, M.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Tritt, A.</dc:creator>
<dc:creator>Oliver Ruebel, O.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Yatsenko, D.</dc:creator>
<dc:creator>Chu, J.</dc:creator>
<dc:creator>Kemere, C.</dc:creator>
<dc:creator>Garcia, S.</dc:creator>
<dc:creator>Buccino, A.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.577295</dc:identifier>
<dc:title><![CDATA[Spyglass: a data analysis framework for reproducible and shareable neuroscience research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.24.577129v1?rss=1">
<title>
<![CDATA[
IgG hexamers initiate acute lung injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.24.577129v1?rss=1</link>
<description><![CDATA[
Antibodies can initiate lung injury in a variety of disease states such as autoimmunity, transfusion reactions, or after organ transplantation, but the key factors determining in vivo pathogenicity of injury-inducing antibodies are unclear. A previously overlooked step in complement activation by IgG antibodies has been elucidated involving interactions between IgG Fc domains that enable assembly of IgG hexamers, which can optimally activate the complement cascade. Here, we tested the in vivo relevance of IgG hexamers in a complement-dependent alloantibody model of acute lung injury. We used three approaches to block alloantibody hexamerization (antibody carbamylation, the K439E Fc mutation, or treatment with domain B from Staphylococcal protein A), all of which reduced acute lung injury. Conversely, Fc mutations promoting spontaneous hexamerization made a harmful alloantibody into a more potent inducer of acute lung injury and rendered an innocuous alloantibody pathogenic. Treatment with a recombinant Fc hexamer  decoy therapeutic protected mice from lung injury, including in a model with transgenic human FCGR2A expression that exacerbated pathology. These results indicate a direct in vivo role of IgG hexamerization in initiating acute lung injury and the potential for therapeutics that inhibit or mimic hexamerization to treat antibody-mediated diseases.

Brief summaryIgG antibodies can form hexamers. This study shows that hexamer assembly is an important event determining the ability of IgG to trigger acute lung injury.

Graphical abstract

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]]></description>
<dc:creator>Cleary, S. J.</dc:creator>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>Tian, J. J.</dc:creator>
<dc:creator>Kwaan, N.</dc:creator>
<dc:creator>Bulkley, D. P.</dc:creator>
<dc:creator>Bentlage, A. E.</dc:creator>
<dc:creator>Vidarsson, G.</dc:creator>
<dc:creator>Boilard, E.</dc:creator>
<dc:creator>Spirig, R.</dc:creator>
<dc:creator>Zimring, J. C.</dc:creator>
<dc:creator>Looney, M. R.</dc:creator>
<dc:date>2024-01-27</dc:date>
<dc:identifier>doi:10.1101/2024.01.24.577129</dc:identifier>
<dc:title><![CDATA[IgG hexamers initiate acute lung injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.30.577940v1?rss=1">
<title>
<![CDATA[
Field evaluation of a volatile pyrethroid spatial repellent and etofenprox-treated clothing for outdoor protection against forest malaria vectors in Cambodia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.30.577940v1?rss=1</link>
<description><![CDATA[
Cambodias goal to eliminate malaria by 2025 is challenged by persisting transmission in the countrys forest and forest fringe areas. People living in, or traveling to the forest, are exposed to malaria vector bites during the day due to Anopheles daytime biting; and during the night, due to low bed net use and open sleeping structures. Volatile pyrethroid spatial repellents (VPSRs), and insecticide treated clothing (ITC) may help address these gaps in protection. In this field study the authors evaluated the outdoor application of one passive, transfluthrin-based VPSR, four etofenprox-ITCs paired with a picaridin topical repellent, and a combination of VPSR and ITC against wild Anopheles landing in Cambodia. Mathematical modeling was also used to predict the reduction of vectorial capacity of these interventions.

A 7x7 Latin-square (6 interventions and one control) was conducted over 49 collection nights in seven temporary, open structures in a forest in Mondulkiri Province, Cambodia. Pairs of participants conducted human landing catches (HLCs) from 18h00 to 06h00, with each collector conducting collections for six hours. A randomly selected subset of collected Anopheles were identified to species using molecular methods. The rate ratio of each intervention compared to the control on Anopheles landings was estimated using a mixed-effect negative binomial regression with intervention, structure, and collector-pair as fixed-effects, and with collection date and structure-night as random effects. The modeling assessment aims to predict the relative reduction in vectoral capacity. Initial calculations involved establishing a "baseline scenario" without intervention, utilizing biometric parameters for Anopheles dirus. Various scenarios accounting for intervention coverage and adherence were then considered. The study aims to update parameters using field study estimates for wild Anopheles, incorporating multiple semi-field estimates for interventions and accounting for the variability and uncertainty in parameter values.

Of the total 8,294 Anopheles specimens collected, 15% (n=1,242) of specimens were confirmed to species or species group via PCR. Fifteen species were confirmed; Anopheles dirus Form A was predominant (n=429), followed by Anopheles maculatus (n=189), and Anopheles minimus (n=60). All six interventions reduced Anopheles landing substantially; protective efficacies ranged between 61% (95% confidence interval (CI): 48 - 71%) (etofenprox-ITC, washed) and 95% (95% CI: 93 - 96%) (combined VPSR and unwashed etofenprox-ITC). Finally, the modelling assessment demonstrates significant reductions in vectoral capacity, with the highest impact observed for the combined ITC and VPSR as well as the VPSR used alone, although effectiveness decreases with intervention aging, and variability exists in the magnitude of predicted reductions due to differences in experimental conditions.

These transfluthrin-based VPSR and etofenprox ITC interventions have the potential to reduce outdoor and daytime Anopheles biting by providing substantial protection against Anopheles landing. One or more of these tools may play a valuable role in the push for elimination in Cambodia and the Greater Mekong Subregion if programs can achieve effective coverage.
]]></description>
<dc:creator>Vajda, E. A.</dc:creator>
<dc:creator>Ross, A.</dc:creator>
<dc:creator>Doum, D.</dc:creator>
<dc:creator>Fairbanks, E.</dc:creator>
<dc:creator>Chitnis, N. A.</dc:creator>
<dc:creator>Hii, J.</dc:creator>
<dc:creator>Moore, S. J.</dc:creator>
<dc:creator>Richardson, J.</dc:creator>
<dc:creator>Macdonald, M.</dc:creator>
<dc:creator>Sovannaroth, S.</dc:creator>
<dc:creator>Kimheng, P.</dc:creator>
<dc:creator>McIver, D. J.</dc:creator>
<dc:creator>Tatarsky, A.</dc:creator>
<dc:creator>Lobo, N. F.</dc:creator>
<dc:date>2024-02-02</dc:date>
<dc:identifier>doi:10.1101/2024.01.30.577940</dc:identifier>
<dc:title><![CDATA[Field evaluation of a volatile pyrethroid spatial repellent and etofenprox-treated clothing for outdoor protection against forest malaria vectors in Cambodia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.02.578712v1?rss=1">
<title>
<![CDATA[
Sex Differences in the Brain's White Matter Microstructure during Development assessed using Advanced Diffusion MRI Models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.02.578712v1?rss=1</link>
<description><![CDATA[
Typical sex differences in white matter (WM) microstructure during development are incompletely understood. Here we evaluated sex differences in WM microstructure during typical brain development using a sample of neurotypical individuals across a wide developmental age (N=239, aged 5-22 years). We used the conventional diffusion-weighted MRI (dMRI) model, diffusion tensor imaging (DTI), and two advanced dMRI models, the tensor distribution function (TDF) and neurite orientation dispersion density imaging (NODDI) to assess WM microstructure. WM microstructure exhibited significant, regionally consistent sex differences across the brain during typical development. Additionally, the TDF model was most sensitive in detecting sex differences. These findings highlight the importance of considering sex in neurodevelopmental research and underscore the value of the advanced TDF model.
]]></description>
<dc:creator>Benavidez, S. M.</dc:creator>
<dc:creator>Abaryan, Z.</dc:creator>
<dc:creator>Kim, G. S.</dc:creator>
<dc:creator>Laltoo, E.</dc:creator>
<dc:creator>McCracken, J. T.</dc:creator>
<dc:creator>Thompson, P. M.</dc:creator>
<dc:creator>Lawrence, K. E.</dc:creator>
<dc:date>2024-02-04</dc:date>
<dc:identifier>doi:10.1101/2024.02.02.578712</dc:identifier>
<dc:title><![CDATA[Sex Differences in the Brain's White Matter Microstructure during Development assessed using Advanced Diffusion MRI Models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.01.578324v1?rss=1">
<title>
<![CDATA[
MAGIC matrices: freeform bioprinting materials to support complex and reproducible organoid morphogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.01.578324v1?rss=1</link>
<description><![CDATA[
Complex and robust self-organization requires defined initial conditions and dynamic boundaries - neighboring tissues and extracellular matrix (ECM) that actively evolve to guide morphogenesis. A major challenge in tissue engineering is identifying material properties that mimic dynamic tissue boundaries but that are compatible with the engineering tools necessary for controlling the initial conditions of culture. Here we describe a highly tunable granular biomaterial, MAGIC matrix, that supports long-term bioprinting and gold-standard tissue self-organization. MAGIC matrix is designed for two temperature regimes: at 4 {degrees}C it exhibits reversible yield-stress behavior to support hours-long high-fidelity 3D printing without compromising cell viability; when transferred to cell culture at 37 {degrees}C, the material cross-links and exhibits viscoelasticity and stress relaxation that can be tuned to match numerous conditions, including that of reconstituted basement membrane matrices like Matrigel. We demonstrate that the timescale of stress relaxation and loss tangent are decoupled in MAGIC matrices, allowing us to test the role of stress relaxation rate and strain-dependence across formulations with identical storage and loss moduli. We find that fast absolute stress relaxation rates and large relative deformation magnitudes are required to optimize for morphogenesis. We apply optimized MAGIC matrices toward precise extrusion bioprinting of saturated cell suspensions directly into 3D culture. The ability to carefully control initial conditions for tissue growth yields dramatic increases in organoid reproducibility and complexity across multiple tissue types. We also fabricate perfusable 3D microphysiological systems that experience large strains in response to pressurization due to the compliant and dynamic tissue boundaries. Combined, our results both identify key parameters for optimal organoid morphogenesis in an engineered material and lay the foundation for fabricating more complex and reproducible tissue morphologies by canalizing their self-organization in both space and time.
]]></description>
<dc:creator>Graham, A. J.</dc:creator>
<dc:creator>Khoo, M. W. L.</dc:creator>
<dc:creator>Srivastava, V.</dc:creator>
<dc:creator>Viragova, S.</dc:creator>
<dc:creator>Parekh, K.</dc:creator>
<dc:creator>Morley, C. D.</dc:creator>
<dc:creator>Bird, M.</dc:creator>
<dc:creator>Lebel, P.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Gomez-Sjoberg, R.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:date>2024-02-05</dc:date>
<dc:identifier>doi:10.1101/2024.02.01.578324</dc:identifier>
<dc:title><![CDATA[MAGIC matrices: freeform bioprinting materials to support complex and reproducible organoid morphogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.05.578995v1?rss=1">
<title>
<![CDATA[
A TBX5-dependent compartment boundary patterns the cardiac interventricular septum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.05.578995v1?rss=1</link>
<description><![CDATA[
Failure of septation of the interventricular septum (IVS) is the most common congenital heart defect (CHD), but mechanisms for patterning the IVS are largely unknown. Here, we show that a Tbx5+/Mef2cAHF+ progenitor lineage forms a compartment boundary bisecting the IVS. This coordinated population originates at a first- and second heart field interface. Ablation of Tbx5+/Mef2cAHF+ progenitors cause IVS disorganization, right ventricular hypoplasia and mixing of IVS lineages. Reduced dosage of the CHD transcription factor TBX5 disrupts boundary position and integrity, resulting in ventricular septation defects (VSDs) and patterning defects, including misexpression of Slit2 and Ntn1, which encode guidance cues. Reducing NTN1 dosage partly rescues cardiac defects in Tbx5 mutant embryos. Loss of Slit2 or Ntn1 causes VSDs and perturbed septal lineage distributions. Thus, we identify Tbx5 as a candidate selector gene, directing progenitors and regulating essential cues, to pattern a compartment boundary for proper cardiac septation, revealing mechanisms for cardiac birth defects.
]]></description>
<dc:creator>Kathiriya, I. S.</dc:creator>
<dc:creator>Dominguez, M. H.</dc:creator>
<dc:creator>Rao, K. S.</dc:creator>
<dc:creator>Muncie-Vasic, J. M.</dc:creator>
<dc:creator>Devine, W. P.</dc:creator>
<dc:creator>Hu, K. M.</dc:creator>
<dc:creator>Hota, S. K.</dc:creator>
<dc:creator>Garay, B. I.</dc:creator>
<dc:creator>Quintero, D.</dc:creator>
<dc:creator>Goyal, P.</dc:creator>
<dc:creator>Matthews, M. N.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Sukonnik, T.</dc:creator>
<dc:creator>Miguel-Perez, D.</dc:creator>
<dc:creator>Winchester, S.</dc:creator>
<dc:creator>Brower, E. F.</dc:creator>
<dc:creator>Forjaz, A.</dc:creator>
<dc:creator>Wu, P.-H.</dc:creator>
<dc:creator>Wirtz, D.</dc:creator>
<dc:creator>Kiemen, A. L.</dc:creator>
<dc:creator>Bruneau, B. G.</dc:creator>
<dc:date>2024-02-05</dc:date>
<dc:identifier>doi:10.1101/2024.02.05.578995</dc:identifier>
<dc:title><![CDATA[A TBX5-dependent compartment boundary patterns the cardiac interventricular septum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.06.579149v1?rss=1">
<title>
<![CDATA[
Protein-protein interactions with G3BPs drive stress granule condensation and gene expression changes under cellular stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.06.579149v1?rss=1</link>
<description><![CDATA[
Stress granules (SGs) are macromolecular assemblies that form under cellular stress. Formation of these condensates is driven by the condensation of RNA and RNA-binding proteins such as G3BPs. G3BPs condense into SGs following stress-induced translational arrest. Three G3BP paralogs (G3BP1, G3BP2A, and G3BP2B) have been identified in vertebrates. However, the contribution of different G3BP paralogs to stress granule formation and stress-induced gene expression changes is incompletely understood. Here, we identified key residues for G3BP condensation such as V11. This conserved amino acid is required for formation of the G3BP-Caprin-1 complex, hence promoting SG assembly. Total RNA sequencing and ribosome profiling revealed that disruption of G3BP condensation corresponds to changes in mRNA levels and ribosome engagement during the integrated stress response (ISR). Moreover, we found that G3BP2B preferentially condenses and promotes changes in mRNA expression under endoplasmic reticulum (ER) stress. Together, this work suggests that stress granule assembly promotes changes in gene expression under cellular stress, which is differentially regulated by G3BP paralogs.
]]></description>
<dc:creator>Liboy-Lugo, J. M.</dc:creator>
<dc:creator>Espinoza, C. A.</dc:creator>
<dc:creator>Sheu-Gruttadauria, J.</dc:creator>
<dc:creator>Park, J. E.</dc:creator>
<dc:creator>Xu, A.</dc:creator>
<dc:creator>Jowhar, Z.</dc:creator>
<dc:creator>Gao, A. L.</dc:creator>
<dc:creator>Carmona-Negron, J. A.</dc:creator>
<dc:creator>Wittmann, T.</dc:creator>
<dc:creator>Jura, N.</dc:creator>
<dc:creator>Floor, S. N.</dc:creator>
<dc:date>2024-02-07</dc:date>
<dc:identifier>doi:10.1101/2024.02.06.579149</dc:identifier>
<dc:title><![CDATA[Protein-protein interactions with G3BPs drive stress granule condensation and gene expression changes under cellular stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.08.578953v1?rss=1">
<title>
<![CDATA[
Minimum Effective Dose of Clemastine in a Mouse Model of Preterm White Matter Injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.08.578953v1?rss=1</link>
<description><![CDATA[
BackgroundPreterm white matter injury (PWMI) is the most common cause of brain injury in premature neonates. PWMI involves a differentiation arrest of oligodendrocytes, the myelinating cells of the central nervous system. Clemastine was previously shown to induce oligodendrocyte differentiation and myelination in mouse models of PWMI at a dose of 10 mg/kg/day. The minimum effective dose (MED) of clemastine is unknown. Identification if the MED is essential for maximizing safety and efficacy in neonatal clinical trials. We hypothesized that the MED in neonatal mice is lower than 10 mg/kg/day.

MethodsMouse pups were exposed to normoxia or hypoxia (10% FiO2) from postnatal day 3 (P3) through P10. Vehicle or clemastine fumarate at one of four doses (0.5, 2, 7.5 or 10 mg/kg/day) was given orally to hypoxia-exposed pups. At P14, myelination was assessed by immunohistochemistry and electron microscopy to determine the MED. Clemastine pharmacokinetics were evaluated at steady-state on day 8 of treatment.

ResultsClemastine rescued hypoxia-induced hypomyelination with a MED of 7.5 mg/kg/day. Pharmacokinetic analysis of the MED revealed Cmax 44.0 ng/mL, t1/2 4.6 hours, and AUC24 280.1 ng*hr/mL.

ConclusionBased on these results, myelination-promoting exposures should be achievable with oral doses of clemastine in neonates with PWMI.

Key PointsO_LIPreterm white matter injury (PWMI) is the most common cause of brain injury and cerebral palsy in premature neonates.
C_LIO_LIClemastine, an FDA-approved antihistamine, was recently identified to strongly promote myelination in a mouse model of PWMI and is a possible treatment.
C_LIO_LIThe minimum effective dose in neonatal rodents is unknown and is critical for guiding dose selection and balancing efficacy with toxicity in future clinical trials.
C_LIO_LIWe identified the minimum effective dose of clemastine and the associated pharmacokinetics in a murine chronic hypoxia model of PWMI, paving the way for a future clinical trial in human neonates.
C_LI
]]></description>
<dc:creator>Odell, E. P.</dc:creator>
<dc:creator>Jabassini, N.</dc:creator>
<dc:creator>Schniedewind, B.</dc:creator>
<dc:creator>Pease-Raissi, S. E.</dc:creator>
<dc:creator>Frymoyer, A.</dc:creator>
<dc:creator>Christians, U.</dc:creator>
<dc:creator>Green, A. J.</dc:creator>
<dc:creator>Chan, J. R.</dc:creator>
<dc:creator>Ostrem, B. E. L.</dc:creator>
<dc:date>2024-02-09</dc:date>
<dc:identifier>doi:10.1101/2024.02.08.578953</dc:identifier>
<dc:title><![CDATA[Minimum Effective Dose of Clemastine in a Mouse Model of Preterm White Matter Injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.07.579345v1?rss=1">
<title>
<![CDATA[
Single Nucleotide Polymorphism (SNP) and Antibody-based Cell Sorting (SNACS): A tool for demultiplexing single-cell DNA sequencing data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.07.579345v1?rss=1</link>
<description><![CDATA[
MotivationRecently, single-cell DNA sequencing (scDNA-seq) and multi-modal profiling with the addition of cell-surface antibodies (scDAb-seq) have provided key insights into cancer heterogeneity.

Scaling these technologies across large patient cohorts, however, is cost and time prohibitive. Multiplexing, in which cells from unique patients are pooled into a single experiment, offers a possible solution. While multiplexing methods exist for scRNAseq, accurate demultiplexing in scDNAseq remains an unmet need.

ResultsHere, we introduce SNACS: Single-Nucleotide Polymorphism (SNP) and Antibody-based Cell Sorting. SNACS relies on a combination of patient-level cell-surface identifiers and natural variation in genetic polymorphisms to demultiplex scDNAseq data. We demonstrated the performance of SNACS on a dataset consisting of multi-sample experiments from patients with leukemia where we knew truth from single-sample experiments from the same patients. Using SNACS, accuracy ranged from 0.948 - 0.991 vs 0.552 - 0.934 using demultiplexing methods from the single-cell literature.

Availability ImplementationSNACS is available at https://github.com/olshena/SNACS.



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]]></description>
<dc:creator>Kennedy, V. E.</dc:creator>
<dc:creator>Roy, R.</dc:creator>
<dc:creator>Peretz, C.</dc:creator>
<dc:creator>Koh, A.</dc:creator>
<dc:creator>Tran, E.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Olshen, A.</dc:creator>
<dc:date>2024-02-10</dc:date>
<dc:identifier>doi:10.1101/2024.02.07.579345</dc:identifier>
<dc:title><![CDATA[Single Nucleotide Polymorphism (SNP) and Antibody-based Cell Sorting (SNACS): A tool for demultiplexing single-cell DNA sequencing data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.09.579451v1?rss=1">
<title>
<![CDATA[
Biochemical, biophysical, and structural investigations of two mutants (C154Y and R312H) of the human Kir2.1 channel involved in the Andersen-Tawil syndrome. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.09.579451v1?rss=1</link>
<description><![CDATA[
Inwardly rectifying potassium (Kir) channels play a pivotal role in physiology by establishing, maintaining, and regulating the resting membrane potential of the cells, particularly contributing to the cellular repolarization of many excitable cells. Dysfunction in Kir2.1 channels is implicated in several chronic and debilitating human diseases for which there are currently no effective treatments. Specifically, Kir2.1-R312H and Kir2.1-C154Y mutations are associated with Andersen-Tawil syndrome (ATS) in humans. We have investigated the impact of these two mutants in the trafficking of the channel to the cell membrane and function in Xenopus laevis oocytes. Despite both mutations being successfully trafficked to the cell membrane and capable of binding PIP2 (phosphatidylinositol-4,5- bisphosphate), the main modulator for channel activity, they resulted in defective channels that do not display K+ current, albeit through different molecular mechanisms. Co-expression studies showed that R312H and C154Y are expressed and associated with the WT subunits. While WT subunits could rescue R312H dysfunction, the presence of a unique C154Y subunit disrupts the function of the entire complex, which is a typical feature of mutations with a dominant-negative effect. Molecular dynamics simulations showed that Kir2.1-C154Y mutation induces a loss in the structural plasticity of the selectivity filter, impairing the K+ flow. In addition, the cryo-EM structure of the Kir2.1-R312H mutant has been reconstructed. This study identified the molecular mechanisms by which two ATS-causing mutations impact Kir2.1 channel function and provide valuable insights that can guide potential strategies for the development of future therapeutic interventions for ATS.
]]></description>
<dc:creator>Zuniga, D.</dc:creator>
<dc:creator>Zoumpoulakis, A.</dc:creator>
<dc:creator>Veloso, R. F.</dc:creator>
<dc:creator>Peverini, L.</dc:creator>
<dc:creator>Shi, S.</dc:creator>
<dc:creator>Pozza, A.</dc:creator>
<dc:creator>Kugler, V.</dc:creator>
<dc:creator>Bonnete, F.</dc:creator>
<dc:creator>Bouceba, T.</dc:creator>
<dc:creator>Wagner, R.</dc:creator>
<dc:creator>Corringer, P.-J.</dc:creator>
<dc:creator>Fernandes, C. A.</dc:creator>
<dc:creator>Venien-Bryan, C.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.09.579451</dc:identifier>
<dc:title><![CDATA[Biochemical, biophysical, and structural investigations of two mutants (C154Y and R312H) of the human Kir2.1 channel involved in the Andersen-Tawil syndrome.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.09.579529v1?rss=1">
<title>
<![CDATA[
Alterations in Lysosomal, Glial and Neurodegenerative Biomarkers in Patients with Sporadic and Genetic Forms of Frontotemporal Dementia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.09.579529v1?rss=1</link>
<description><![CDATA[
BackgroundFrontotemporal dementia (FTD) is the most common cause of early-onset dementia with 10-20% of cases caused by mutations in one of three genes: GRN, C9orf72, or MAPT. To effectively develop therapeutics for FTD, the identification and characterization of biomarkers to understand disease pathogenesis and evaluate the impact of specific therapeutic strategies on the target biology as well as the underlying disease pathology are essential. Moreover, tracking the longitudinal changes of these biomarkers throughout disease progression is crucial to discern their correlation with clinical manifestations for potential prognostic usage.

MethodsWe conducted a comprehensive investigation of biomarkers indicative of lysosomal biology, glial cell activation, synaptic and neuronal health in cerebrospinal fluid (CSF) and plasma from non-carrier controls, sporadic FTD (symptomatic non-carriers) and symptomatic carriers of mutations in GRN, C9orf72, or MAPT, as well as asymptomatic GRN mutation carriers. We also assessed the longitudinal changes of biomarkers in GRN mutation carriers. Furthermore, we examined biomarker levels in disease impacted brain regions including middle temporal gyrus (MTG) and superior frontal gyrus (SFG) and disease-unaffected inferior occipital gyrus (IOG) from sporadic FTD and symptomatic GRN carriers.

ResultsWe confirmed glucosylsphingosine (GlcSph), a lysosomal biomarker regulated by progranulin, was elevated in the plasma from GRN mutation carriers, both symptomatic and asymptomatic. GlcSph and other lysosomal biomarkers such as ganglioside GM2 and globoside GB3 were increased in the disease affected SFG and MTG regions from sporadic FTD and symptomatic GRN mutation carriers, but not in the IOG, compared to the same brain regions from controls. The glial biomarkers GFAP in plasma and YKL40 in CSF were elevated in asymptomatic GRN carriers, and all symptomatic groups, except the symptomatic C9orf72 mutation group. YKL40 was also increased in SFG and MTG regions from sporadic FTD and symptomatic GRN mutation carriers. Neuronal injury and degeneration biomarkers NfL in CSF and plasma, and UCHL1 in CSF were elevated in patients with all forms of FTD. Synaptic biomarkers NPTXR, NPTX1/2, and VGF were reduced in CSF from patients with all forms of FTD, with the most pronounced reductions observed in symptomatic MAPT mutation carriers. Furthermore, we demonstrated plasma NfL was significantly positively correlated with disease severity as measured by CDR+NACC FTLD{square}SB in genetic forms of FTD and CSF NPTXR was significantly negatively correlated with CDR+NACC FTLD{square}SB in symptomatic GRN and MAPT mutation carriers.

ConclusionsIn conclusion, our comprehensive investigation replicated alterations in biofluid biomarkers indicative of lysosomal function, glial activation, synaptic and neuronal health across sporadic and genetic forms of FTD and unveiled novel insights into the dysregulation of these biomarkers within brain tissues from patients with GRN mutations. The observed correlations between biomarkers and disease severity open promising avenues for prognostic applications and for indicators of drug efficacy in clinical trials. Our data also implicated a complicated relationship between biofluid and tissue biomarker changes and future investigations should delve into the mechanistic underpinnings of these biomarkers, which will serve as a foundation for the development of targeted therapeutics for FTD.
]]></description>
<dc:creator>Hsiao-Nakamoto, J.</dc:creator>
<dc:creator>Chiu, C.-L.</dc:creator>
<dc:creator>VandeVrede, L.</dc:creator>
<dc:creator>Ravi, R.</dc:creator>
<dc:creator>Vandenberg, B.</dc:creator>
<dc:creator>De Groot, J.</dc:creator>
<dc:creator>Tsogtbaatar, B.</dc:creator>
<dc:creator>Fang, M.</dc:creator>
<dc:creator>Auger, P.</dc:creator>
<dc:creator>Gould, N. S.</dc:creator>
<dc:creator>Marchioni, F.</dc:creator>
<dc:creator>Powers, C. A.</dc:creator>
<dc:creator>Davis, S. S.</dc:creator>
<dc:creator>Suh, J. H.</dc:creator>
<dc:creator>Alkabsh, J.</dc:creator>
<dc:creator>Heuer, H. W.</dc:creator>
<dc:creator>Lago, A. L.</dc:creator>
<dc:creator>Scearce-Levie, K.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Boeve, B. F.</dc:creator>
<dc:creator>Rosen, H. J.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:creator>Tsai, R.</dc:creator>
<dc:creator>Di Paolo, G.</dc:creator>
<dc:creator>Boxer, A. L.</dc:creator>
<dc:creator>Bhalla, A.</dc:creator>
<dc:creator>Huang, F.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.09.579529</dc:identifier>
<dc:title><![CDATA[Alterations in Lysosomal, Glial and Neurodegenerative Biomarkers in Patients with Sporadic and Genetic Forms of Frontotemporal Dementia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.12.578709v1?rss=1">
<title>
<![CDATA[
Intercalated amygdala dysfunction drives extinction deficits in the Sapap3 mouse model of obsessive-compulsive disorder 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.12.578709v1?rss=1</link>
<description><![CDATA[
BackgroundThe avoidance of aversive stimuli due to negative reinforcement learning is critical for survival in real-world environments, which demand dynamic responding to both positive and negative stimuli that often conflict with each other. Individuals with obsessive-compulsive disorder (OCD) commonly exhibit impaired negative reinforcement and extinction, perhaps involving deficits in amygdala functioning. An amygdala subregion of particular interest is the intercalated nuclei of the amygdala (ITC) which has been linked to negative reinforcement and extinction, with distinct clusters mediating separate aspects of behavior. This study focuses on the dorsal ITC cluster (ITCd) and its role in negative reinforcement during a complex behavior that models real-world dynamic decision making.

MethodsWe investigated the impact of ITCd function on negative reinforcement and extinction by applying fiber photometry measurement of GCamp6f signals and optogenetic manipulations during a platform-mediated avoidance task in a mouse model of OCD-like behavior: the Sapap3-null mouse.

ResultsWe find impaired neural activity in the ITCd of male and female Sapap3-null mice to the encoding of negative stimuli during platform-mediated avoidance. Sapap3-null mice also exhibit deficits in extinction of avoidant behavior, which is modulated by ITCd neural activity.

ConclusionsSapap3-null mice fail to extinguish avoidant behavior in platform-mediated avoidance, due to heightened ITCd activity. This deficit can be rescued by optogenetically inhibiting ITCd during extinction. Together, our results provide insight into the neural mechanisms underpinning negative reinforcement deficits in the context of OCD, emphasizing the necessity of ITCd in responding to negative stimuli in complex environments.
]]></description>
<dc:creator>St. Laurent, R.</dc:creator>
<dc:creator>Kusche, K. M.</dc:creator>
<dc:creator>Kreitzer, A. C.</dc:creator>
<dc:creator>Malenka, R. C.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.12.578709</dc:identifier>
<dc:title><![CDATA[Intercalated amygdala dysfunction drives extinction deficits in the Sapap3 mouse model of obsessive-compulsive disorder]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.15.580326v1?rss=1">
<title>
<![CDATA[
Prosaposin is cleaved into saposins by multiple cathepsins in a progranulin-regulated fashion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.15.580326v1?rss=1</link>
<description><![CDATA[
Prosaposin (PSAP) is a lysosomal protein that plays a key role in sphingolipid metabolism. PSAP is cleaved into four bioactive disulfide-rich peptides, saposins A, B, C, and D, that catalyze sphin-golipidases to promote sphingolipid breakdown. Considering the key role of PSAP and saposins in sphingolipid metabolism and the existence of genetic mutations in PSAP associated with juve-nile-onset lysosomal storage and adult-onset neurodegenerative diseases, maintaining optimal levels of PSAP and saposins is crucial for proper lysosomal function and sphingolipid homeosta-sis. Despite this, the mechanism by which saposins are released from PSAP, and thus available to modulate sphingolipidases, sphingolipid homeostasis, and downstream lysosomal function, is not well understood. Here, we performed a comprehensive study to identify lysosomal enzymes which regulate prosaposin cleavage into saposins. In vitro cleavage assays identified multiple enzymes that can process human prosaposin into multi- and single-saposin fragments in a pH-dependent manner. We confirmed the role of cathepsins D and B in PSAP processing and showed that these and several previously unidentified lysosomal proteases (cathepsins E, K, L, S, V, G and AEP/LGMN) are able to process PSAP in distinctive, pH-dependent manners. In addition, we have demonstrated a novel role for progranulin (PGRN) in the regulation of PSAP cleavage. We found that PGRN and multi-granulin fragments (MGFs) directly regulate the cleavage of PSAP by cathepsin D. With this study, we have identified that multiple cathepsins, PGRN and MGFs work in concert to produce saposins under different conditions, which could present novel opportunities to modulate saposin levels in disease.
]]></description>
<dc:creator>Hodul, M.</dc:creator>
<dc:creator>Lane-Donovan, C.</dc:creator>
<dc:creator>Mambou, E. A.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Kao, A. W.</dc:creator>
<dc:date>2024-02-16</dc:date>
<dc:identifier>doi:10.1101/2024.02.15.580326</dc:identifier>
<dc:title><![CDATA[Prosaposin is cleaved into saposins by multiple cathepsins in a progranulin-regulated fashion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.16.580396v1?rss=1">
<title>
<![CDATA[
Degron-based bioPROTACs for controlling signaling in CAR T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.16.580396v1?rss=1</link>
<description><![CDATA[
Chimeric antigen receptor (CAR) T cells have made a tremendous impact in the clinic, but potent signaling through the CAR can be detrimental to treatment safety and efficacy. The use of protein degradation to control CAR signaling can address these issues in pre-clinical models. Existing strategies for regulating CAR stability rely on small molecules to induce systemic degradation. In contrast to small molecule regulation, genetic circuits offer a more precise method to control CAR signaling in an autonomous, cell-by-cell fashion. Here, we describe a programmable protein degradation tool that adopts the framework of bioPROTACs, heterobifunctional proteins that are composed of a target recognition domain fused to a domain that recruits the endogenous ubiquitin proteasome system. We develop novel bioPROTACs that utilize a compact four residue degron and demonstrate degradation of cytosolic and membrane protein targets using either a nanobody or synthetic leucine zipper as a protein binder. Our bioPROTACs exhibit potent degradation of CARs and can inhibit CAR signaling in primary human T cells. We demonstrate the utility of our bioPROTACs by constructing a genetic circuit to degrade the tyrosine kinase ZAP70 in response to recognition of a specific membrane-bound antigen. This circuit is able to disrupt CAR T cell signaling only in the presence of a specific cell population. These results suggest that bioPROTACs are a powerful tool for expanding the cell engineering toolbox for CAR T cells.
]]></description>
<dc:creator>Kim, M. S.</dc:creator>
<dc:creator>Bhargava, H. K.</dc:creator>
<dc:creator>Shavey, G. E.</dc:creator>
<dc:creator>Lim, W. A.</dc:creator>
<dc:creator>El-Samad, H.</dc:creator>
<dc:creator>Ng, A. H.</dc:creator>
<dc:date>2024-02-17</dc:date>
<dc:identifier>doi:10.1101/2024.02.16.580396</dc:identifier>
<dc:title><![CDATA[Degron-based bioPROTACs for controlling signaling in CAR T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.14.580363v1?rss=1">
<title>
<![CDATA[
High-resolution single particle imaging at 100-200 keV with the Gatan Alpine direct electron detector. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.14.580363v1?rss=1</link>
<description><![CDATA[
Developments in direct electron detector technology have played a pivotal role in enabling high-resolution structural studies by cryo-EM at 200 and 300 keV. Yet, theory and recent experiments indicate advantages to imaging at 100 keV, energies for which the current detectors have not been optimized. In this study, we evaluated the Gatan Alpine detector, designed for operation at 100 and 200 keV. Compared to the Gatan K3, Alpine demonstrated a significant DQE improvement at these voltages, specifically a ~4-fold improvement at Nyquist at 100 keV. In single-particle cryo-EM experiments, Alpine datasets yielded better than 2 [A] resolution reconstructions of apoferritin at 120 and 200 keV on a ThermoFisher Scientific (TFS) Glacios microscope. We also achieved a ~3.2 [A] resolution reconstruction for a 115 kDa asymmetric protein complex, proving its effectiveness with complex biological samples. In-depth analysis revealed that Alpine reconstructions are comparable to K3 reconstructions at 200 keV, and remarkably, reconstruction from Alpine at 120 keV on a TFS Glacios surpassed all but the 300 keV data from a TFS Titan Krios with GIF/K3. Additionally, we show Alpines capability for high-resolution data acquisition and screening on lower-end systems by obtaining ~3 [A] resolution reconstructions of apoferritin and aldolase at 100 keV and detailed 2D averages of a 55 kDa sample using a side-entry cryo holder. Overall, we show that Gatan Alpine performs well with the standard 200 keV imaging systems and may potentially capture the benefits of lower accelerating voltages, possibly bringing smaller sized particles within the scope of cryo-EM.
]]></description>
<dc:creator>Chan, L. M.</dc:creator>
<dc:creator>Courteau, B. J.</dc:creator>
<dc:creator>Maker, A.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Basanta, B.</dc:creator>
<dc:creator>Mehmood, H.</dc:creator>
<dc:creator>Bulkley, D.</dc:creator>
<dc:creator>Joyce, D.</dc:creator>
<dc:creator>Lee, B. C.</dc:creator>
<dc:creator>Mick, S.</dc:creator>
<dc:creator>Gulati, S.</dc:creator>
<dc:creator>Lander, G. C.</dc:creator>
<dc:creator>Verba, K. A.</dc:creator>
<dc:date>2024-02-18</dc:date>
<dc:identifier>doi:10.1101/2024.02.14.580363</dc:identifier>
<dc:title><![CDATA[High-resolution single particle imaging at 100-200 keV with the Gatan Alpine direct electron detector.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.14.580356v1?rss=1">
<title>
<![CDATA[
Brain substates induced by DMT relate to sympathetic output and meaningfulness of the experience 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.14.580356v1?rss=1</link>
<description><![CDATA[
N,N-Dimethyltryptamine (DMT) is a serotonergic psychedelic, known to rapidly induce short-lasting alterations in conscious experience, characterized by a profound and immersive sense of physical transcendence alongside rich and vivid auditory distortions and visual imagery. Multimodal neuroimaging data paired with dynamic analysis techniques offer a valuable approach for identifying unique signatures of brain activity - and linked autonomic physiology - naturally unfolding during the altered state of consciousness induced by DMT. We leveraged simultaneous fMRI and EKG data acquired in 14 healthy volunteers prior to, during, and after intravenous administration of DMT, and, separately, placebo. fMRI data was preprocessed to derive individual dynamic activity matrices, reflecting the similarity of brain activity in time, and community detection algorithms were applied on these matrices to identify brain activity substates; EKG data was used to derive continuous heart rate. We identified a brain substate occurring immediately after DMT injection, characterized by hippocampal and medial parietal deactivations and increased superior temporal lobe activity under DMT. Deactivations in the hippocampus and medial parietal cortex correlated with alterations in the usual sense of time, space and self-referential processes, reflecting a deconstruction of essential features of ordinary consciousness. Superior lobe activations instead correlated with audio/visual hallucinations and experience of "entities", reflecting the emergence of altered sensory experiences under DMT. Finally, increased heart rate under DMT correlated positively with hippocampus/medial parietal deactivation and the experience of "entities", and negatively with altered self-referential processes. These results suggest a chain of influence linking sympathetic regulation to hippocampal and medial parietal deactivations under DMT, which combined, may contribute to positive mental health outcomes related to self-referential processing following psychedelic administration.
]]></description>
<dc:creator>Pasquini, L.</dc:creator>
<dc:creator>Simon, A.</dc:creator>
<dc:creator>Gallen, C.</dc:creator>
<dc:creator>Kettner, H.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Gazzaley, A.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Timmermann, C.</dc:creator>
<dc:date>2024-02-19</dc:date>
<dc:identifier>doi:10.1101/2024.02.14.580356</dc:identifier>
<dc:title><![CDATA[Brain substates induced by DMT relate to sympathetic output and meaningfulness of the experience]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.17.580803v1?rss=1">
<title>
<![CDATA[
Activated STING in the thymus alters T cell development and selection leading to autoimmunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.17.580803v1?rss=1</link>
<description><![CDATA[
Classifying systemic inflammatory disorders as autoinflammatory or autoimmune provides insight into disease pathogenesis and whether treatment should target innate molecules and their signaling pathways or the adaptive immune response. COPA syndrome is a monogenic disorder of immune dysregulation that leads to interstitial lung disease and high-titer autoantibodies. Studies show constitutive activation of the innate immune molecule STING is centrally involved in disease. However, the mechanisms by which STING results in loss of T cell tolerance and autoimmunity in COPA syndrome or more common autoimmune diseases is not understood. Using CopaE241K/+ mice, we uncovered a functional role for STING in the thymus. Single cell data of human thymus demonstrates STING is highly expressed in medullary thymic epithelial cells (mTECs) involved in processing and presenting self-antigens to thymocytes. In CopaE241K/+ mice, activated STING in mTECs triggered interferon signaling, impaired macroautophagy and caused a defect in negative selection of T cells. Wild-type mice given a systemic STING agonist phenocopied the selection defect and showed enhanced thymic escape of a T cell clone targeting a self-antigen also expressed in melanoma. Our work demonstrates STING activation in TECs shapes the T cell repertoire and contributes to autoimmunity, findings important for settings that activate thymic STING.

SummaryActivation of STING in thymic epithelial cells shifts the T cell repertoire to promote autoimmunity providing a new mechanistic link between innate and adaptive immunity.
]]></description>
<dc:creator>Deng, Z.</dc:creator>
<dc:creator>Law, C. S.</dc:creator>
<dc:creator>Kurra, S.</dc:creator>
<dc:creator>Simchoni, N.</dc:creator>
<dc:creator>Shum, A. K.</dc:creator>
<dc:date>2024-02-19</dc:date>
<dc:identifier>doi:10.1101/2024.02.17.580803</dc:identifier>
<dc:title><![CDATA[Activated STING in the thymus alters T cell development and selection leading to autoimmunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.19.581062v1?rss=1">
<title>
<![CDATA[
Inhibition of Indirect Pathway Activity Causes Abnormal Decision-Making In a Mouse Model of Impulse Control Disorder in Parkinson's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.19.581062v1?rss=1</link>
<description><![CDATA[
Healthy action selection relies on the coordinated activity of striatal direct and indirect pathway neurons. In Parkinsons disease (PD), in which loss of midbrain dopamine neurons is associated with progressive motor and cognitive deficits, this coordination is disrupted. Dopamine replacement therapy can remediate motor symptoms, but can also cause impulse control disorder (ICD), which is characterized by pathological gambling, hypersexuality, and/or compulsive shopping. The cellular and circuit mechanisms of ICD remain unknown. Here we developed a mouse model of PD/ICD, in which ICD-like behavior was assayed with a delay discounting task. We found that in parkinsonian mice, the dopamine agonist pramipexole drove more pronounced delay discounting, as well as disrupted firing in both direct and indirect pathway neurons. We found that chemogenetic inhibition of indirect pathway neurons in parkinsonian mice drove similar phenotypes. Together, these findings provide a new mouse model and insights into ICD pathophysiology.
]]></description>
<dc:creator>Zhuang, X.</dc:creator>
<dc:creator>Lemak, J.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2024-02-22</dc:date>
<dc:identifier>doi:10.1101/2024.02.19.581062</dc:identifier>
<dc:title><![CDATA[Inhibition of Indirect Pathway Activity Causes Abnormal Decision-Making In a Mouse Model of Impulse Control Disorder in Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.20.580754v1?rss=1">
<title>
<![CDATA[
Microglia are not required for maintenance of blood-brain barrier properties in health, but PLX5622 alters brain endothelial cholesterol metabolism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.20.580754v1?rss=1</link>
<description><![CDATA[
Microglia are resident immune cells of the central nervous system, yet their functions far exceed those related to immunology. From pruning neural synapses during development to preventing excessive neural activity throughout life, microglia are intimately involved in the brains most basic processes. Studies have reported a close interaction between microglia and endothelial cells, as well as both helpful and harmful roles for microglia at the blood-brain barrier (BBB) in the context of disease. However, much less work has been done to understand microglia-endothelial cell interactions in the healthy brain. Here, we aim to determine the role of microglia at the healthy BBB. We used the colony-stimulating factor 1 receptor (CSF1R) inhibitor PLX5622 to deplete microglia and analyzed BBB ultrastructure, permeability, and transcriptome. Interestingly, we found that, despite their direct contact with endothelial cells, microglia are not necessary for maintenance of BBB structure, function, or gene expression in the healthy brain. However, we found that PLX5622 treatment alters brain endothelial cholesterol metabolism, and this effect was independent from microglial depletion, suggesting PLX5622 has off-target effects on brain vasculature.
]]></description>
<dc:creator>Profaci, C. P.</dc:creator>
<dc:creator>Harvey, S. S.</dc:creator>
<dc:creator>Bajc, K.</dc:creator>
<dc:creator>Zhang, T. Z.</dc:creator>
<dc:creator>Zhang, A. Z.</dc:creator>
<dc:creator>Nemec, K. M.</dc:creator>
<dc:creator>McKinsey, G. L.</dc:creator>
<dc:creator>Longworth, A.</dc:creator>
<dc:creator>McMullen, T. P.</dc:creator>
<dc:creator>Arnold, T. D.</dc:creator>
<dc:creator>Lawson, D. A.</dc:creator>
<dc:creator>Bennett, F. C.</dc:creator>
<dc:creator>Davalos, D.</dc:creator>
<dc:creator>Daneman, R.</dc:creator>
<dc:date>2024-02-22</dc:date>
<dc:identifier>doi:10.1101/2024.02.20.580754</dc:identifier>
<dc:title><![CDATA[Microglia are not required for maintenance of blood-brain barrier properties in health, but PLX5622 alters brain endothelial cholesterol metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.21.581282v1?rss=1">
<title>
<![CDATA[
Fetal brain response to maternal inflammation requires microglia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.21.581282v1?rss=1</link>
<description><![CDATA[
In utero infection and maternal inflammation can adversely impact fetal brain development. Maternal systemic illness, even in the absence of direct fetal central nervous system infection, is associated with an increased risk of autism and schizophrenia in affected offspring. The cell types mediating the response of the fetal brain to maternal inflammation are largely unknown, hindering the development of therapies to prevent and treat adverse neuropsychiatric outcomes. Here, we show that microglia, the resident phagocytes of the brain, are enriched for expression of receptors for relevant pathogens and cytokines, throughout embryonic development. Using a rodent maternal immune activation (MIA) model in which polyinosinic:polycytidylic acid is injected into pregnant dams, we demonstrate long-lasting transcriptional changes in fetal microglia that persist into postnatal life. We find that MIA induces widespread gene expression changes in neuronal and non-neuronal cells; importantly, these responses are abolished by selective genetic deletion of microglia, indicating that microglia are required for the transcriptional response of other cortical cell types to MIA. These findings demonstrate that microglia play a critical, durable role in fetal response to maternal inflammation, pointing at microglia as a potential therapeutic cell target.
]]></description>
<dc:creator>Ostrem, B.</dc:creator>
<dc:creator>Dominguez Iturza, N.</dc:creator>
<dc:creator>Stogsdill, J.</dc:creator>
<dc:creator>Faits, T.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Levin, J. Z.</dc:creator>
<dc:creator>Arlotta, P.</dc:creator>
<dc:date>2024-02-22</dc:date>
<dc:identifier>doi:10.1101/2024.02.21.581282</dc:identifier>
<dc:title><![CDATA[Fetal brain response to maternal inflammation requires microglia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.20.580732v1?rss=1">
<title>
<![CDATA[
Superoxide enters neurons via LRRC8A-containing volume-regulated anion channels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.20.580732v1?rss=1</link>
<description><![CDATA[
Superoxide (O2-) is both an intercellular signaling molecule and a cause of neuronal oxidative stress. Superoxide entry into neurons is thought to be indirect, requiring its dismutation to nonpolar hydrogen peroxide. Here we show instead that superoxide enters neurons directly, via LRRC8A-containing volume-sensitive organic anion channels. In primary cultures, neuronal oxidative stress induced either by NMDA receptor stimulation or exposure to authentic superoxide was blocked by the anion channel blockers DIDS and DCPIB and by LRRC8A gene disruption. In mouse cortex, neuronal oxidative stress induced by either NMDA injection or transient ischemia was likewise blocked by both DCPIB and LRRC8A gene disruption. These findings identify a role for LRRC8A-containing volume-sensitive organic anion channels in neuronal oxidative signaling, stress, and glutamate excitotoxicity.
]]></description>
<dc:creator>Harris, K.</dc:creator>
<dc:creator>Uruk, G.</dc:creator>
<dc:creator>Won, S. J.</dc:creator>
<dc:creator>Mai, N.</dc:creator>
<dc:creator>Baxter, P.</dc:creator>
<dc:creator>Everaerts, K.</dc:creator>
<dc:creator>Sah, R.</dc:creator>
<dc:creator>Swanson, R. A.</dc:creator>
<dc:date>2024-02-23</dc:date>
<dc:identifier>doi:10.1101/2024.02.20.580732</dc:identifier>
<dc:title><![CDATA[Superoxide enters neurons via LRRC8A-containing volume-regulated anion channels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.23.581689v1?rss=1">
<title>
<![CDATA[
Molecular and Cellular Mechanisms of Teneurin Signaling in Synaptic Partner Matching 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.23.581689v1?rss=1</link>
<description><![CDATA[
In developing brains, axons exhibit remarkable precision in selecting synaptic partners among many non-partner cells. Evolutionally conserved teneurins were the first identified transmembrane proteins that instruct synaptic partner matching. However, how intracellular signaling pathways execute teneurins functions is unclear. Here, we use in situ proximity labeling to obtain the intracellular interactome of teneurin (Ten-m) in the Drosophila brain. Genetic interaction studies using quantitative partner matching assays in both olfactory receptor neurons (ORNs) and projection neurons (PNs) reveal a common pathway: Ten-m binds to and negatively regulates a RhoGAP, thus activating the Rac1 small GTPases to promote synaptic partner matching. Developmental analyses with single-axon resolution identify the cellular mechanism of synaptic partner matching: Ten-m signaling promotes local F-actin levels and stabilizes ORN axon branches that contact partner PN dendrites. Combining spatial proteomics and high-resolution phenotypic analyses, this study advanced our understanding of both cellular and molecular mechanisms of synaptic partner matching.

HIGHLIGHTSO_LIIn situ spatial proteomics reveal the first intracellular interactome of teneurins
C_LIO_LITen-m signals via a RhoGAP and Rac1 GTPase to regulate synaptic partner matching
C_LIO_LISingle-axon analyses reveal a stabilization-upon-contact model for partner matching
C_LIO_LITen-m signaling promotes F-actin in axon branches contacting partner dendrites
C_LI
]]></description>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Lyu, C.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>McLaughlin, C.</dc:creator>
<dc:creator>Wong, K. K. L.</dc:creator>
<dc:creator>Xie, Q.</dc:creator>
<dc:creator>Luginbuhl, D. J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Udeshi, N.</dc:creator>
<dc:creator>Svinkina, T.</dc:creator>
<dc:creator>Mani, D. R.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Han, S.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Guajardo, R.</dc:creator>
<dc:creator>Ting, A. Y.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Luo, L.</dc:creator>
<dc:date>2024-02-23</dc:date>
<dc:identifier>doi:10.1101/2024.02.23.581689</dc:identifier>
<dc:title><![CDATA[Molecular and Cellular Mechanisms of Teneurin Signaling in Synaptic Partner Matching]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.28.582543v1?rss=1">
<title>
<![CDATA[
PIKfyve controls dendritic cell function and tumor immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.28.582543v1?rss=1</link>
<description><![CDATA[
The modern armamentarium for cancer treatment includes immunotherapy and targeted therapy, such as protein kinase inhibitors. However, the mechanisms that allow cancer-targeting drugs to effectively mobilize dendritic cells (DCs) and affect immunotherapy are poorly understood. Here, we report that among shared gene targets of clinically relevant protein kinase inhibitors, high PIKFYVE expression was least predictive of complete response in patients who received immune checkpoint blockade (ICB). In immune cells, high PIKFYVE expression in DCs was associated with worse response to ICB. Genetic and pharmacological studies demonstrated that PIKfyve ablation enhanced DC function via selectively altering the alternate/non-canonical NF-{kappa}B pathway. Both loss of Pikfyve in DCs and treatment with apilimod, a potent and specific PIKfyve inhibitor, restrained tumor growth, enhanced DC-dependent T cell immunity, and potentiated ICB efficacy in tumor-bearing mouse models. Furthermore, the combination of a vaccine adjuvant and apilimod reduced tumor progression in vivo. Thus, PIKfyve negatively controls DCs, and PIKfyve inhibition has promise for cancer immunotherapy and vaccine treatment strategies.
]]></description>
<dc:creator>Choi, J. E.</dc:creator>
<dc:creator>Qiao, Y.</dc:creator>
<dc:creator>Kryczek, I.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Gurkan, J.</dc:creator>
<dc:creator>Bao, Y.</dc:creator>
<dc:creator>Gondal, M.</dc:creator>
<dc:creator>Tien, J. C.-Y.</dc:creator>
<dc:creator>Maj, T.</dc:creator>
<dc:creator>Yazdani, S.</dc:creator>
<dc:creator>Parolia, A.</dc:creator>
<dc:creator>Xia, H.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Wei, S.</dc:creator>
<dc:creator>Grove, S.</dc:creator>
<dc:creator>Vatan, L.</dc:creator>
<dc:creator>Lin, H.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Su, F.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>He, T.</dc:creator>
<dc:creator>Cieslik, M.</dc:creator>
<dc:creator>Green, M. D.</dc:creator>
<dc:creator>Zou, W.</dc:creator>
<dc:creator>Chinnaiyan, A. M.</dc:creator>
<dc:date>2024-03-02</dc:date>
<dc:identifier>doi:10.1101/2024.02.28.582543</dc:identifier>
<dc:title><![CDATA[PIKfyve controls dendritic cell function and tumor immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.04.583400v1?rss=1">
<title>
<![CDATA[
A Spatial Multi-Modal Dissection of Host-Microbiome Interactions within the Colitis Tissue Microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.04.583400v1?rss=1</link>
<description><![CDATA[
The intricate and dynamic interactions between the host immune system and its microbiome constituents undergo dynamic shifts in response to perturbations to the intestinal tissue environment. Our ability to study these events on the systems level is significantly limited by in situ approaches capable of generating simultaneous insights from both host and microbial communities. Here, we introduce Microbiome Cartography (MicroCart), a framework for simultaneous in situ probing of host features and its microbiome across multiple spatial modalities. We demonstrate MicroCart by comprehensively investigating the alterations in both gut host and microbiome components in a murine model of colitis by coupling MicroCart with spatial proteomics, transcriptomics, and glycomics platforms. Our findings reveal a global but systematic transformation in tissue immune responses, encompassing tissue-level remodeling in response to host immune and epithelial cell state perturbations, and bacterial population shifts, localized inflammatory responses, and metabolic process alterations during colitis. MicroCart enables a deep investigation of the intricate interplay between the host tissue and its microbiome with spatial multiomics.
]]></description>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Bai, Y.</dc:creator>
<dc:creator>Yeo, Y. Y.</dc:creator>
<dc:creator>Lu, X.</dc:creator>
<dc:creator>Rovira-Clave, X.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Yeung, J.</dc:creator>
<dc:creator>Gerber, G. K.</dc:creator>
<dc:creator>Angelo, M.</dc:creator>
<dc:creator>Shalek, A. K.</dc:creator>
<dc:creator>Nolan, G. P.</dc:creator>
<dc:creator>Jiang, S.</dc:creator>
<dc:date>2024-03-06</dc:date>
<dc:identifier>doi:10.1101/2024.03.04.583400</dc:identifier>
<dc:title><![CDATA[A Spatial Multi-Modal Dissection of Host-Microbiome Interactions within the Colitis Tissue Microenvironment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.05.583578v1?rss=1">
<title>
<![CDATA[
Regulation of virion production by the ORF8 signal peptide across SARS-CoV-2 variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.05.583578v1?rss=1</link>
<description><![CDATA[
The open reading frame 8 (ORF8), an accessory protein of SARS-CoV-2, is prone to deletions and mutations across different viral variants, which was first described in several Singapore variants. The reason why viral evolution favors loss or inactivation of ORF8 is not fully understood, although the effects of ORF8 on inflammation, immune evasion, and disease severity have been described. Here we show -using clinical ORF8-deficient viral isolates, virus-like particles (VLPs) and viral replicons- that ORF8 expression dampens viral particle production. ORF8 physically interacts with the viral Spike protein and induces Golgi fragmentation, overall contributing to less virus particle production. Using systematic ORF8 deletions, we mapped the particle-reducing function to its N-terminal signal peptide. Interestingly, this part of ORF8 is severely truncated in the recent XBB.1.5 variant, and when restored, suppresses viral particle production in the context of the entire viral genome. Collectively, our data supports the model that evolutionary pressure exists to delete ORF8 sequence and expression across SARS-CoV-2 variants to fully enable viral particle production.

ImportanceSARS-CoV-2 variants continue to emerge worldwide with advantages in replication and immune evasion. Many variants have acquired distinct mutations in independent lineages to abolish ORF8 expression. To understand the molecular mechanisms behind this evolutionary trend, we utilized reverse genetics, molecular virology, and confocal microscopy to show that ORF8 has antiviral functions by dampening viral particle production and inducing Golgi stress during infection. Our data demonstrate that SARS-CoV-2 is continuing its adaptation to optimize viral particle production and other unknown aspects of viral infection.
]]></description>
<dc:creator>Khalid, M. M.</dc:creator>
<dc:creator>Chen, I.</dc:creator>
<dc:creator>Soveg, F. S.</dc:creator>
<dc:creator>Taha, T. Y.</dc:creator>
<dc:creator>Tabata, T.</dc:creator>
<dc:creator>Suryawanshi, R.</dc:creator>
<dc:creator>Syed, A. M.</dc:creator>
<dc:creator>Ciling, A.</dc:creator>
<dc:creator>McCavitt-Malvido, M.</dc:creator>
<dc:creator>Schulze-Gahmen, U.</dc:creator>
<dc:creator>Hayashi, J.</dc:creator>
<dc:creator>Kim, I.-J.</dc:creator>
<dc:creator>Fong, S. W.</dc:creator>
<dc:creator>Batra, J.</dc:creator>
<dc:creator>Kumar, G. R.</dc:creator>
<dc:creator>Laurent, R.</dc:creator>
<dc:creator>NG, L. F.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:creator>Verdin, E.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:date>2024-03-06</dc:date>
<dc:identifier>doi:10.1101/2024.03.05.583578</dc:identifier>
<dc:title><![CDATA[Regulation of virion production by the ORF8 signal peptide across SARS-CoV-2 variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.05.583634v1?rss=1">
<title>
<![CDATA[
Rare Variable M. tuberculosis Antigens induce predominant Th17 responses in human infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.05.583634v1?rss=1</link>
<description><![CDATA[
CD4 T cells are essential for immunity to M. tuberculosis (Mtb), and emerging evidence indicates that IL-17-producing Th17 cells contribute to immunity to Mtb. While identifying protective T cell effector functions is important for TB vaccine design, T cell antigen specificity is also likely to be important. To identify antigens that induce protective immunity, we reasoned that as in other pathogens, effective immune recognition drives sequence diversity in individual Mtb antigens. We previously identified Mtb genes under evolutionary diversifying selection pressure whose products we term Rare Variable Mtb Antigens (RVMA). Here, in two distinct human cohorts with recent exposure to TB, we found that RVMA preferentially induce CD4 T cells that express RoR{gamma}t and produce IL-17, in contrast to  classical Mtb antigens that induce T cells that produce IFN{gamma}. Our results suggest that RVMA can be valuable antigens in vaccines for those already infected with Mtb to amplify existing antigen-specific Th17 responses to prevent TB disease.
]]></description>
<dc:creator>Ogongo, P.</dc:creator>
<dc:creator>Wassie, L.</dc:creator>
<dc:creator>Tran, A.</dc:creator>
<dc:creator>Columbus, D.</dc:creator>
<dc:creator>Sharling, L.</dc:creator>
<dc:creator>Ouma, G.</dc:creator>
<dc:creator>Ouma, S. G.</dc:creator>
<dc:creator>Bobosha, K.</dc:creator>
<dc:creator>Lindestam Arlehamn, C. S.</dc:creator>
<dc:creator>Gandhi, N. R.</dc:creator>
<dc:creator>Auld, S. C.</dc:creator>
<dc:creator>Rengarajan, J.</dc:creator>
<dc:creator>Day, C. L.</dc:creator>
<dc:creator>Altman, J. D.</dc:creator>
<dc:creator>Blumberg, H.</dc:creator>
<dc:creator>Ernst, J. D.</dc:creator>
<dc:date>2024-03-06</dc:date>
<dc:identifier>doi:10.1101/2024.03.05.583634</dc:identifier>
<dc:title><![CDATA[Rare Variable M. tuberculosis Antigens induce predominant Th17 responses in human infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.06.583756v1?rss=1">
<title>
<![CDATA[
CD47 is Required for Mesenchymal Progenitor Proliferation and Fracture Repair 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.06.583756v1?rss=1</link>
<description><![CDATA[
CD47 is a ubiquitous and pleiotropic cell-surface receptor. Disrupting CD47 enhances injury repair in various tissues but the role of CD47 has not been studied in bone injuries. In a murine closed-fracture model, CD47-null mice showed decreased callus bone volume, bone mineral content, and tissue mineral content as assessed by microcomputed tomography 10 days post-fracture, and increased fibrous volume as determined by histology. To understand the cellular basis for this phenotype, mesenchymal progenitors (MSC) were harvested from bone marrow. CD47-null MSC showed decreased large fibroblast colony formation (CFU-F), significantly less proliferation, and fewer cells in S-phase, although osteoblast differentiation was unaffected. However, consistent with prior research, CD47-null endothelial cells showed increased proliferation relative to WT cells. Similarly, in a murine ischemic fracture model, CD47-null mice showed reduced fracture callus bone volume and bone mineral content relative to WT. Consistent with our in vitro results, in vivo EdU labeling showed decreased cell proliferation in the callus of CD47-null mice, while staining for CD31 and endomucin demonstrated increased endothelial cell mass. Finally, WT mice administered a CD47 morpholino, which blocks CD47 protein production, showed a callus phenotype similar to that of non-ischemic and ischemic fractures in CD47-null mice, suggesting the phenotype was not due to developmental changes in the knockout mice. Thus, inhibition of CD47 during bone healing reduces both non-ischemic and ischemic fracture healing, in part, by decreasing MSC proliferation. Furthermore, the increase in endothelial cell proliferation and early blood vessel density caused by CD47 disruption is not sufficient to overcome MSC dysfunction.
]]></description>
<dc:creator>Zondervan, R. L.</dc:creator>
<dc:creator>Capobianco, C. A.</dc:creator>
<dc:creator>Jenkins, D. C.</dc:creator>
<dc:creator>Reicha, J. D.</dc:creator>
<dc:creator>Fredrick, L. M.</dc:creator>
<dc:creator>Lam, C.</dc:creator>
<dc:creator>Isenberg, J. S.</dc:creator>
<dc:creator>Ahn, J.</dc:creator>
<dc:creator>Marcucio, R. S.</dc:creator>
<dc:creator>Hankenson, K. D.</dc:creator>
<dc:date>2024-03-06</dc:date>
<dc:identifier>doi:10.1101/2024.03.06.583756</dc:identifier>
<dc:title><![CDATA[CD47 is Required for Mesenchymal Progenitor Proliferation and Fracture Repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.04.583403v1?rss=1">
<title>
<![CDATA[
Selective vulnerability and resilience to Alzheimer's disease tauopathy as a function of genes and the connectome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.04.583403v1?rss=1</link>
<description><![CDATA[
Brain regions in Alzheimers (AD) exhibit distinct vulnerability to the diseases hallmark pathology, with the entorhinal cortex and hippocampus succumbing early to tau tangles while others like primary sensory cortices remain resilient. The quest to understand how local/regional genetic factors, pathogenesis, and network-mediated spread of pathology together govern this selective vulnerability (SV) or resilience (SR) is ongoing. Although many risk genes in AD are known from gene association and transgenic studies, it is still not known whether and how their baseline expression signatures confer SV or SR to brain structures. Prior analyses have yielded conflicting results, pointing to a disconnect between the location of genetic risk factors and downstream tau pathology. We hypothesize that a full accounting of genes role in mediating SV/SR would require the modeling of network-based vulnerability, whereby tau misfolds, aggregates, and propagates along fiber projections.

We therefore employed an extended network diffusion model (eNDM) and tested it on tau pathology PET data from 196 AD patients from the Alzheimers Disease Neuroimaging Initiative (ADNI). Thus the fitted eNDM model becomes a reference process from which to assess the role of innate genetic factors. Using the residual (observed - model-predicted) tau as a novel target outcome, we obtained its association with 100 top AD risk-genes, whose baseline spatial transcriptional profiles were obtained from the Allen Human Brain Atlas (AHBA). We found that while many risk genes at baseline showed a strong association with regional tau, many more showed a stronger association with residual tau. This suggests that both direct vulnerability, related to the network, as well as network-independent vulnerability, are conferred by risk genes. We then classified risk genes into four classes: network-related SV (SV-NR), network-independent SV (SV-NI), network-related SR (SR-NR), and network-independent SR (SR-NI). Each class has a distinct spatial signature and associated vulnerability to tau. Remarkably, we found from gene-ontology analyses, that genes in these classes were enriched in distinct functional processes and encompassed different functional networks. These findings offer new insights into the factors governing innate vulnerability or resilience in AD pathophysiology and may prove helpful in identifying potential intervention targets.
]]></description>
<dc:creator>Anand, C.</dc:creator>
<dc:creator>Torok, J.</dc:creator>
<dc:creator>Abdelnour, F.</dc:creator>
<dc:creator>Maia, P.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:date>2024-03-08</dc:date>
<dc:identifier>doi:10.1101/2024.03.04.583403</dc:identifier>
<dc:title><![CDATA[Selective vulnerability and resilience to Alzheimer's disease tauopathy as a function of genes and the connectome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.06.583752v1?rss=1">
<title>
<![CDATA[
Lack of TGFβ signaling competency predicts immune poor cancer conversion to immune rich and response to checkpoint blockade 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.06.583752v1?rss=1</link>
<description><![CDATA[
BackgroundThe efficacy of immune checkpoint blockade (ICB) depends on restoring immune recognition of cancer cells that have evaded immune surveillance. At the time of diagnosis, patients with lymphocyte-infiltrated cancers are the most responsive to ICB, yet a considerable fraction of patients have immune-poor tumors.

MethodsWe analyzed transcriptomic data from IMvigor210, TCGA, and TISMO datasets to evaluate the predictive value of {beta}Alt, a score representing the negative correlation of signatures consisting of transforming growth factor beta (TGF{beta}) targets and genes involved in error-prone DNA repair. The immune context of {beta}Alt was assessed by evaluating tumor-educated immune signatures. An ICB-resistant, high {beta}Alt preclinical tumor model was treated with a TGF{beta} inhibitor, radiation, and/or ICB and assessed for immune composition and tumor control.

ResultsHere, we show that high {beta}Alt is associated with an immune-poor context yet is predictive of ICB response in both humans and mice. A high {beta}Alt cancer in which TGF{beta} signaling is compromised generates a TGF{beta} rich, immunosuppressive tumor microenvironment. Accordingly, preclinical modeling showed that TGF{beta} inhibition followed by radiotherapy could convert an immune-poor, ICB-resistant tumor to an immune-rich, ICB-responsive tumor. Mechanistically, TGF{beta} blockade in irradiated tumors activated natural killer cells that were required to recruit lymphocytes to respond to ICB. In support of this, natural killer cell activation signatures were also increased in immune-poor mouse and human tumors that responded to ICB.

ConclusionsThese studies suggest that loss of TGF{beta} competency identifies a subset of cold tumors that are candidates for ICB. Our mechanistic studies show that inhibiting TGF{beta} activity converts high {beta}Alt, cold tumors into ICB-responsive tumors via NK cells. Thus, a biomarker consisting of combined TGF{beta}, DNA repair, and immune context signatures provides a means to prospectively identify patients whose cancers may be converted from  cold to  hot, which could be exploited for therapeutic treatment. O_LIWhat is already known on this topic - For some cancer patients, response to ICB provides durable tumor control. Current biomarkers are insufficient to reliably predict the immunotherapy response for most patients, particularly those whose tumors lack lymphocytic infiltration.
C_LIO_LIWhat this study adds - The {beta}Alt score, which reports a DNA damage deficiency caused by lack of TGF{beta} signaling, predicts response to ICB in clinical trial data from IMvigor210 metastatic bladder cancer patients and for melanoma patients. Notably, transcriptomic assessment of the immune context shows that these are immune-poor, so-called "cold" tumors. Preclinical modeling indicates that alleviating TGF{beta} inhibition of NK cells is critical to relieving immunosuppression.
C_LIO_LIHow this study might affect research, practice or policy - Our work identifies a novel tumor phenotype consisting of cancer cells that have lost TGF{beta} signaling and gained error-prone DNA repair embedded in a TGF{beta} rich, immune-poor microenvironment, which is conserved across cancer types in humans and among preclinical tumor models. Patients whose immune-poor tumors have high {beta}Alt scores are strong candidates for ICB and radiotherapy combinations that may be further augmented by TGF{beta} inhibition. Hence, the {beta}Alt score can be used to stratify immune-poor cancer patients for optimal therapeutic strategies.
C_LI
]]></description>
<dc:creator>Moore, J.</dc:creator>
<dc:creator>Gkantalis, J.</dc:creator>
<dc:creator>Guix, I.</dc:creator>
<dc:creator>Chou, W.</dc:creator>
<dc:creator>Yuen, K.</dc:creator>
<dc:creator>Lazar, A.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:creator>combes, a.</dc:creator>
<dc:creator>Barcellos-Hoff, M. H.</dc:creator>
<dc:date>2024-03-08</dc:date>
<dc:identifier>doi:10.1101/2024.03.06.583752</dc:identifier>
<dc:title><![CDATA[Lack of TGFβ signaling competency predicts immune poor cancer conversion to immune rich and response to checkpoint blockade]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.08.583956v1?rss=1">
<title>
<![CDATA[
Combining Cell-Intrinsic and -Extrinsic Resistance to HIV-1 By Engineering Hematopoietic Stem Cells for CCR5 Knockout and B Cell Secretion of Therapeutic Antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.08.583956v1?rss=1</link>
<description><![CDATA[
Autologous transplantation of CCR5 null hematopoietic stem and progenitor cells (HSPCs) is the only known cure for HIV-1 infection. However, this treatment is limited because of the rarity of CCR5-null matched donors, the morbidities associated with allogeneic transplantation, and the prevalence of HIV-1 strains resistant to CCR5 knockout (KO) alone. Here, we propose a one-time therapy through autologous transplantation of HSPCs genetically engineered ex vivo to produce both CCR5 KO cells and long-term secretion of potent HIV-1 inhibiting antibodies from B cell progeny. CRISPR-Cas9-engineered HSPCs maintain engraftment capacity and multi-lineage potential in vivo and can be engineered to express multiple antibodies simultaneously. Human B cells engineered to express each antibody secrete neutralizing concentrations capable of inhibiting HIV-1 pseudovirus infection in vitro. This work lays the groundwork for a potential one-time functional cure for HIV-1 through combining the long-term delivery of therapeutic antibodies against HIV-1 and the known efficacy of CCR5 KO HSPC transplantation.
]]></description>
<dc:creator>Feist, W. N.</dc:creator>
<dc:creator>Luna, S. E.</dc:creator>
<dc:creator>Ben-Efraim, K.</dc:creator>
<dc:creator>Filsinger Interrante, M. V.</dc:creator>
<dc:creator>Amorin, N. A.</dc:creator>
<dc:creator>Johnston, N. M.</dc:creator>
<dc:creator>Bruun, T. U. J.</dc:creator>
<dc:creator>Ghanim, H. Y.</dc:creator>
<dc:creator>Lesch, B. J.</dc:creator>
<dc:creator>Dudek, A. M.</dc:creator>
<dc:creator>Porteus, M. H.</dc:creator>
<dc:date>2024-03-09</dc:date>
<dc:identifier>doi:10.1101/2024.03.08.583956</dc:identifier>
<dc:title><![CDATA[Combining Cell-Intrinsic and -Extrinsic Resistance to HIV-1 By Engineering Hematopoietic Stem Cells for CCR5 Knockout and B Cell Secretion of Therapeutic Antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.05.583407v1?rss=1">
<title>
<![CDATA[
Understanding the complex interplay between tau, amyloid and the network in the spatiotemporal progression of Alzheimer's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.05.583407v1?rss=1</link>
<description><![CDATA[
INTRODUCTIONThe interaction of amyloid and tau in neurodegenerative diseases is a central feature of AD pathophysiology. While experimental studies point to various interaction mechanisms, their causal direction and mode (local, remote or network-mediated) remain unknown in human subjects. The aim of this study was to compare mathematical reaction-diffusion models encoding distinct cross-species couplings to identify which interactions were key to model success.

METHODSWe tested competing mathematical models of network spread, aggregation, and amyloid-tau interactions on publicly available data from ADNI.

RESULTSAlthough network spread models captured the spatiotemporal evolution of tau and amyloid in human subjects, the model including a one-way amyloid-to-tau aggregation interaction performed best.

DISCUSSIONThis mathematical exposition of the "pas de deux" of co-evolving proteins provides quantitative, whole-brain support to the concept of amyloid-facilitated-tauopathy rather than the classic amyloid-cascade or pure-tau hypotheses, and helps explain certain known but poorly understood aspects of AD.
]]></description>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Torok, J.</dc:creator>
<dc:date>2024-03-11</dc:date>
<dc:identifier>doi:10.1101/2024.03.05.583407</dc:identifier>
<dc:title><![CDATA[Understanding the complex interplay between tau, amyloid and the network in the spatiotemporal progression of Alzheimer's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.08.584173v1?rss=1">
<title>
<![CDATA[
Cell type-specific dynamics of prefrontal gamma synchrony during flexible behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.08.584173v1?rss=1</link>
<description><![CDATA[
Cognitive dysfunction in conditions such as schizophrenia involves disrupted communication between the prefrontal cortex (PFC) and mediodorsal thalamus (MD). Parvalbumin interneurons (PVI) are known to regulate PFC microcircuits and generate gamma-frequency ([~]40Hz) oscillations - fast, synchronized neural rhythms that are recruited during many executive functions, necessary for cognitive flexibility, and deficient in schizophrenia. While targeting PVI-mediated gamma oscillations holds great therapeutic promise, their nature and specific functions, e.g., for regulating PFC[-&gt;]MD communication, remain elusive. Using dual-color voltage indicators and optogenetics, we reveal that PVIs dynamically entrain MD-projecting PFC neurons both locally and contralaterally, giving rise to multiple distinct circuit-specific patterns of distributed synchronization that are recruited in a behaviorally-specific manner to support particular aspects of flexible behavior. Thus, gamma oscillations are not unitary phenomena characterized by one microcircuit-wide pattern of entrainment. Rather, they comprise diverse motifs, defined by specific cell types and phase relationships, that are dynamically recruited for specific functions.
]]></description>
<dc:creator>Phensy, A. J.</dc:creator>
<dc:creator>Hagopian, L. L.</dc:creator>
<dc:creator>Sohal, V. S.</dc:creator>
<dc:date>2024-03-11</dc:date>
<dc:identifier>doi:10.1101/2024.03.08.584173</dc:identifier>
<dc:title><![CDATA[Cell type-specific dynamics of prefrontal gamma synchrony during flexible behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.11.584505v1?rss=1">
<title>
<![CDATA[
Bridging Domains in Chronic Lower Back Pain: Large Language Models and Ontology-driven Strategies for Knowledge Graph Construction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.11.584505v1?rss=1</link>
<description><![CDATA[
Link prediction and entity resolution play pivotal roles in uncovering hidden relationships within networks and ensuring data quality in the era of heterogeneous data integration. This paper explores the utilization of large language models to enhance link prediction, particularly through knowledge graphs derived from transdisciplinary literature. Investigating zero-shot entity resolution techniques, we examine the impact of ontology-based and large language model approaches on the stability of link prediction results. Through a case study focusing on chronic lower back pain research, we analyze workflow decisions and their influence on prediction outcomes. Our research underscores the importance of robust methodologies in improving predictive accuracy and data integration across diverse domains.
]]></description>
<dc:creator>Anderson, P. E.</dc:creator>
<dc:creator>Davidson, J. M.</dc:creator>
<dc:creator>Lin, D.</dc:creator>
<dc:creator>Migler, T.</dc:creator>
<dc:creator>Ho, I.</dc:creator>
<dc:creator>Koenig, C.</dc:creator>
<dc:creator>Bittner, M.</dc:creator>
<dc:creator>Kaplan, S.</dc:creator>
<dc:creator>Paraiso, M.</dc:creator>
<dc:creator>Buhn, N.</dc:creator>
<dc:creator>Stokes, E.</dc:creator>
<dc:creator>Hunt, T.</dc:creator>
<dc:creator>Ropella, G.</dc:creator>
<dc:creator>Lotz, J.</dc:creator>
<dc:date>2024-03-14</dc:date>
<dc:identifier>doi:10.1101/2024.03.11.584505</dc:identifier>
<dc:title><![CDATA[Bridging Domains in Chronic Lower Back Pain: Large Language Models and Ontology-driven Strategies for Knowledge Graph Construction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.12.584554v1?rss=1">
<title>
<![CDATA[
Bayesian inference of state feedback control parameters for fo perturbation responses in cerebellar ataxia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.12.584554v1?rss=1</link>
<description><![CDATA[
Behavioral speech tasks have been widely used to understand the mechanisms of speech motor control in healthy speakers as well as in various clinical populations. However, determining which neural functions differ between healthy speakers and clinical populations based on behavioral data alone is difficult because multiple mechanisms may lead to the same behavioral differences. For example, individuals with cerebellar ataxia (CA) produce abnormally large compensatory responses to pitch perturbations in their auditory feedback, compared to controls, but this pattern could have many explanations.

Here, computational modeling techniques were used to address this challenge. Bayesian inference was used to fit a state feedback control (SFC) model of voice fundamental frequency (fo) control to the behavioral pitch perturbation responses of individuals with CA and healthy controls. This fitting process resulted in estimates of posterior likelihood distributions of five model parameters (sensory feedback delays, absolute and relative levels of auditory and somatosensory feedback noise, and controller gain), which were compared between the two groups. Results suggest that the CA group may proportionally weight auditory and somatosensory feedback differently from the control group. Specifically, the CA group showed a greater relative sensitivity to auditory feedback than the control group. There were also large group differences in the controller gain parameter, suggesting increased motor output responses to target errors in the CA group. These modeling results generate hypotheses about how CA may affect the speech motor system, which could help guide future empirical investigations in CA. This study also demonstrates the overall proof-of-principle of using this Bayesian inference approach to understand behavioral speech data in terms of interpretable parameters of speech motor control models.

Author summaryCerebellar ataxia is a condition characterized by a loss of coordination in the control of muscle movements, including those required for speech, due to damage in the cerebellar region of the brain. Behavioral speech experiments have been used to understand this disorders impact on speech motor control, but the results can be ambiguous to interpret. In this study, we fit a computational model of the neural speech motor control system to the speech data of individuals with cerebellar ataxia and that of healthy controls to determine what differences in model parameters best explain how the two groups differ in their control of vocal pitch. We found that group differences may be explained by increased sensitivity to auditory feedback prediction errors (differences between the actual sound speakers hear of their own speech as they produce it and the sound they expected to hear) and increased motor response in individuals with cerebellar ataxia. These computational results help us understand how cerebellar ataxia impacts speech motor control, and this general approach can also be applied to study other neurological speech disorders.
]]></description>
<dc:creator>Gaines, J. L.</dc:creator>
<dc:creator>Kim, K. S.</dc:creator>
<dc:creator>Parrell, B.</dc:creator>
<dc:creator>Ramanarayanan, V.</dc:creator>
<dc:creator>Pongos, A. L.</dc:creator>
<dc:creator>Nagarajan, S. S.</dc:creator>
<dc:creator>Houde, J. F.</dc:creator>
<dc:date>2024-03-14</dc:date>
<dc:identifier>doi:10.1101/2024.03.12.584554</dc:identifier>
<dc:title><![CDATA[Bayesian inference of state feedback control parameters for fo perturbation responses in cerebellar ataxia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.12.584638v1?rss=1">
<title>
<![CDATA[
CDK4 and CDK6 upregulation promotes DNA replication stress, genomic instability and resistance to EGFR targeted therapy in lung cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.12.584638v1?rss=1</link>
<description><![CDATA[
Epidermal growth factor receptor (EGFR)-mutant lung adenocarcinomas (LUAD) harbor a complex landscape of genetic co-alterations and potential oncogenic interactions. Among them are recurrent amplifications of the cell cycle regulatory genes CDK4 and CDK6, which have been clinically implicated in resistance to EGFR tyrosine kinase inhibitors (TKIs). However, the mechanisms by which CDK4/6 upregulation promotes therapy resistance remain poorly defined. Here, we demonstrate that CDK4 or CDK6 overexpression limits EGFR inhibitor-induced proliferative arrest, promoting continued cell cycle progression. This is accompanied by elevated replication stress, increased TPX2 expression, DNA damage leading to ATM activation, and ultimately genomic instability. Integrative transcriptomic and copy number analyses of EGFR-mutant LUAD tumors from both patients and preclinical models revealed that CDK4 or CDK6 amplification is associated with the upregulation of genes linked to tumor progression, including AGR2, ASNS, and STEAP1. CDK4 amplification was also highly correlated with gene expression changes associated with epithelial-to-mesenchymal transition (EMT) in a single-cell RNA sequencing dataset from patient biopsies. In preclinical models, co-treatment with CDK4/6 and EGFR inhibitors restored proliferative arrest, induced tumor cell apoptosis, and reduced replication stress, DNA damage, and genomic instability. Our findings uncover a mechanistic basis for EGFR inhibitor resistance of CDK4 and CDK6 amplified EGFR-mutant LUAD. They also provide a rationale for the biomarker-driven clinical development of combination EGFR and CDK4/6-targeted therapies for the treatment of a subset of EGFR-mutant LUAD patients.
]]></description>
<dc:creator>Gini, B.</dc:creator>
<dc:creator>Gui, P.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Kerr, D. L.</dc:creator>
<dc:creator>Tan, L.</dc:creator>
<dc:creator>Barbosa, D.</dc:creator>
<dc:creator>Olivas, V.</dc:creator>
<dc:creator>Gomez, C.</dc:creator>
<dc:creator>Elmes, S.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Chakrabarti, T.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:creator>Blakely, C. M.</dc:creator>
<dc:date>2024-03-14</dc:date>
<dc:identifier>doi:10.1101/2024.03.12.584638</dc:identifier>
<dc:title><![CDATA[CDK4 and CDK6 upregulation promotes DNA replication stress, genomic instability and resistance to EGFR targeted therapy in lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.13.584903v1?rss=1">
<title>
<![CDATA[
Eed controls craniofacial osteoblast differentiation and mesenchymal proliferation from the neural crest 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.13.584903v1?rss=1</link>
<description><![CDATA[
The histone methyltransferase Polycomb repressive complex 2 (PRC2) is required for specification of the neural crest, and mis-regulation of neural crest development can cause severe congenital malformations. PRC2 is necessary for neural crest induction, but the embryonic, cellular, and molecular consequences of PRC2 activity after neural crest induction are incompletely understood. Here we show that Eed, a core subunit of PRC2, is required for craniofacial osteoblast differentiation and mesenchymal proliferation after induction of the neural crest. Integrating mouse genetics with single-cell RNA sequencing and epigenetic profiling, our results reveal that conditional knockout of Eed after neural crest cell induction causes severe craniofacial hypoplasia, impaired craniofacial osteogenesis, and attenuated craniofacial mesenchymal cell proliferation that is first evident in post-migratory neural crest cell populations. We show that Eed drives mesenchymal differentiation and proliferation in vivo and in primary craniofacial cell cultures by epigenetically regulating diverse transcription factor programs that are required for specification of post-migratory neural crest cells. These data enhance understanding of epigenetic mechanisms that underlie craniofacial development, and shed light on the embryonic, cellular, and molecular drivers of rare congenital syndromes in humans.
]]></description>
<dc:creator>Casey-Clyde, T.</dc:creator>
<dc:creator>Liu, S. J.</dc:creator>
<dc:creator>Serrano, J. A. C.</dc:creator>
<dc:creator>Teng, C.</dc:creator>
<dc:creator>Jang, Y.-G.</dc:creator>
<dc:creator>Vasudevan, H. N.</dc:creator>
<dc:creator>Bush, J. O.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:date>2024-03-14</dc:date>
<dc:identifier>doi:10.1101/2024.03.13.584903</dc:identifier>
<dc:title><![CDATA[Eed controls craniofacial osteoblast differentiation and mesenchymal proliferation from the neural crest]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.14.584991v1?rss=1">
<title>
<![CDATA[
5-HT1B receptors mediate dopaminergic inhibition of vesicular fusion and GABA release from striatonigral synapses. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.14.584991v1?rss=1</link>
<description><![CDATA[
The substantia nigra pars reticulata (SNr), a crucial basal ganglia output nucleus, contains a dense expression of dopamine D1 receptors (D1Rs), along with dendrites belonging to dopaminergic neurons of substantia nigra pars compacta. These D1Rs are primarily located on the terminals of striatonigral medium spiny neurons, suggesting their involvement in the regulation of neurotransmitter release from the direct pathway in response to somatodendritic dopamine release. To explore the hypothesis that D1Rs modulate GABA release from striatonigral synapses, we conducted optical recordings of striatonigral activity and postsynaptic patch-clamp recordings from SNr neurons in the presence of dopamine and D1R agonists. We found that dopamine inhibits optogenetically triggered striatonigral GABA release by modulating vesicle fusion and Ca2+ influx in striatonigral boutons. Notably, the effect of DA was independent of D1R activity but required activation of 5-HT1B receptors. Our results suggest a serotonergic mechanism involved in the therapeutic actions of dopaminergic medications for Parkinsons disease and psychostimulant-related disorders.
]]></description>
<dc:creator>Molinari, M.</dc:creator>
<dc:creator>Lieberman, O. J.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Santini, E.</dc:creator>
<dc:creator>Borgkvist, A.</dc:creator>
<dc:date>2024-03-14</dc:date>
<dc:identifier>doi:10.1101/2024.03.14.584991</dc:identifier>
<dc:title><![CDATA[5-HT1B receptors mediate dopaminergic inhibition of vesicular fusion and GABA release from striatonigral synapses.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.18.585609v1?rss=1">
<title>
<![CDATA[
Design principles for accurate folding of DNA origami 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.18.585609v1?rss=1</link>
<description><![CDATA[
We describe design principles for accurate folding of three-dimensional DNA origami. To evaluate design rules, we reduced the problem of DNA strand routing to the known problem of shortest-path finding in a weighted graph. To score candidate DNA strand routes we used a thermodynamic model that accounts for enthalpic and entropic contributions of initial binding, hybridization, and DNA loop closure. We encoded and analyzed new and previously reported design heuristics. Using design principles emerging from this analysis, we redesigned and fabricated multiple shapes and compared their folding accuracy using electrophoretic mobility analysis and electron microscopy imaging. We demonstrate accurate folding can be achieved by optimizing staple routes using our model, and provide a computational framework for applying our methodology to any design.
]]></description>
<dc:creator>Aksel, T.</dc:creator>
<dc:creator>Navarro, E. J.</dc:creator>
<dc:creator>Fong, N.</dc:creator>
<dc:creator>Douglas, S. M.</dc:creator>
<dc:date>2024-03-19</dc:date>
<dc:identifier>doi:10.1101/2024.03.18.585609</dc:identifier>
<dc:title><![CDATA[Design principles for accurate folding of DNA origami]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.19.585567v1?rss=1">
<title>
<![CDATA[
Autonomic Nervous System activity correlates with peak experiences induced by DMT and predicts increases in wellbeing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.19.585567v1?rss=1</link>
<description><![CDATA[
Non-ordinary states of consciousness induced by psychedelics can be accompanied by so-called  peak experiences, characterized at the emotional level by their intensity and positive valence. These experiences are strong predictors of positive outcomes following psychedelic-assisted therapy, and it is therefore important to better understand their biology. Despite growing evidence that the autonomic nervous system (ANS) plays an important role in mediating emotional experiences, its involvement in the psychedelic experience is poorly understood. The aim of this study was to investigate to what extant changes in the relative influence of the sympathetic (SNS) and parasympathetic nervous systems (PNS) over cardiac activity may reflect the subjective experience induced by the short-acting psychedelic N,N-Dimethyltryptamine (DMT). We derived measures of SNS and PNS activity from the electrocardiogram data of 17 participants (11 males, 6 females, mean age = 33.8 y, SD = 8.3) while they received either DMT or placebo. Results show that the joint influence of SNS and PNS ( sympatho-vagal coactivation) over cardiac activity was robustly correlated with participants ratings of  Spiritual Experience and  Insightfulness during the DMT experience, while also being related to improved wellbeing scores two weeks after the session. In addition, we found that the state of balance between the two ANS branches ( sympatho-vagal balance) before DMT injection predicted scores of  Insightfulness during the DMT experience. These findings demonstrate the important involvement of the ANS in psychedelic-induced peak experiences and may pave the way to the development of biofeedback-based tools to enhance psychedelic-therapy.

Significance statementPsychedelics can give rise to intense positive subjective experiences - aligned with Maslows notion of  peak experiences - that can have a positive and enduring impact on mental health. Understanding how these experiences relate to peripheral physiology before and during the acute effects of psychedelics is an important object of enquiry, as it may help advance the therapeutic use of these compounds. In this study, we demonstrate that specific peripheral states computed from heart rate activity recordings predicted and correlated with acute peak experiences and increases in wellbeing. These findings have implications for the relationship between peripheral physiology and altered states of consciousness. Moreover, they highlight a putative marker of physiological  readiness prior the psychedelic experience that could predict therapeutically relevant mechanisms that might be modified to improve mental health outcomes in psychedelic-therapy.
]]></description>
<dc:creator>Bonnelle, V.</dc:creator>
<dc:creator>Feilding, A.</dc:creator>
<dc:creator>Rosas, F. E.</dc:creator>
<dc:creator>Nutt, D. J.</dc:creator>
<dc:creator>Carhart-Harris, R. L.</dc:creator>
<dc:creator>Timmermann, C.</dc:creator>
<dc:date>2024-03-20</dc:date>
<dc:identifier>doi:10.1101/2024.03.19.585567</dc:identifier>
<dc:title><![CDATA[Autonomic Nervous System activity correlates with peak experiences induced by DMT and predicts increases in wellbeing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.19.585648v1?rss=1">
<title>
<![CDATA[
Parallel Encoding of Speech in Human Frontal and Temporal Lobes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.19.585648v1?rss=1</link>
<description><![CDATA[
Models of speech perception are centered around a hierarchy in which auditory representations in the thalamus propagate to primary auditory cortex, then to the lateral temporal cortex, and finally through dorsal and ventral pathways to sites in the frontal lobe. However, evidence for short latency speech responses and low-level spectrotemporal representations in frontal cortex raises the question of whether speech-evoked activity in frontal cortex strictly reflects downstream processing from lateral temporal cortex or whether there are direct parallel pathways from the thalamus or primary auditory cortex to the frontal lobe that supplement the traditional hierarchical architecture. Here, we used high-density direct cortical recordings, high-resolution diffusion tractography, and hemodynamic functional connectivity to evaluate for evidence of direct parallel inputs to frontal cortex from low-level areas. We found that neural populations in the frontal lobe show speech-evoked responses that are synchronous or occur earlier than responses in the lateral temporal cortex. These short latency frontal lobe neural populations encode spectrotemporal speech content indistinguishable from spectrotemporal encoding patterns observed in the lateral temporal lobe, suggesting parallel auditory speech representations reaching temporal and frontal cortex simultaneously. This is further supported by white matter tractography and functional connectivity patterns that connect the auditory nucleus of the thalamus (medial geniculate body) and the primary auditory cortex to the frontal lobe. Together, these results support the existence of a robust pathway of parallel inputs from low-level auditory areas to frontal lobe targets and illustrate long-range parallel architecture that works alongside the classical hierarchical speech network model.
]]></description>
<dc:creator>Hullett, P. W.</dc:creator>
<dc:creator>Leonard, M. K.</dc:creator>
<dc:creator>Gorno-Tempini, M. L. W.</dc:creator>
<dc:creator>Mandelli, M. L.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:date>2024-03-20</dc:date>
<dc:identifier>doi:10.1101/2024.03.19.585648</dc:identifier>
<dc:title><![CDATA[Parallel Encoding of Speech in Human Frontal and Temporal Lobes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.20.585970v1?rss=1">
<title>
<![CDATA[
Chemoproteomics reveals immunogenic and tumor-associated cell surface substrates of ectokinase CK2α 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.20.585970v1?rss=1</link>
<description><![CDATA[
New epitopes for immune recognition provide the basis of anticancer immunity. Due to the high concentration of extracellular adenosine triphosphate in the tumor microenvironment, we hypothesized that extracellular kinases (ectokinases) could have dysregulated activity and introduce aberrant phosphorylation sites on cell surface proteins. We engineered a cell-tethered version of the extracellular kinase CK2, demonstrated it was active on cells under tumor-relevant conditions, and profiled its substrate scope using a chemoproteomic workflow. We then demonstrated that mice developed polyreactive antisera in response to syngeneic tumor cells that had been subjected to surface hyperphosphorylation with CK2. Interestingly, these mice developed B cell and CD4+ T cell responses in response to these antigens but failed to develop a CD8+ T cell response. This work provides a workflow for probing the extracellular phosphoproteome and demonstrates that extracellular phosphoproteins are immunogenic even in a syngeneic system.
]]></description>
<dc:creator>Delaveris, C. S.</dc:creator>
<dc:creator>Kong, S.</dc:creator>
<dc:creator>Glasgow, J.</dc:creator>
<dc:creator>Loudermilk, R. P.</dc:creator>
<dc:creator>Kirkemo, L. L.</dc:creator>
<dc:creator>Zhao, F.</dc:creator>
<dc:creator>Salangsang, F.</dc:creator>
<dc:creator>Phojanakong, P.</dc:creator>
<dc:creator>Serrano, J. A. C.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2024-03-20</dc:date>
<dc:identifier>doi:10.1101/2024.03.20.585970</dc:identifier>
<dc:title><![CDATA[Chemoproteomics reveals immunogenic and tumor-associated cell surface substrates of ectokinase CK2α]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.24.586471v1?rss=1">
<title>
<![CDATA[
Defining the constituents and functions of a lipid-based jumbo phage compartment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.24.586471v1?rss=1</link>
<description><![CDATA[
Viruses are vulnerable to cellular defenses at the start of the infection when a single copy genome has not yet initiated de novo viral protein synthesis. A family of jumbo phages related to phage {Phi}KZ, which infects Pseudomonas aeruginosa, uses a protein-based phage nucleus to protect the DNA during middle and late stages of infection, but how it is protected prior to phage nucleus assembly is unclear. To reveal the environment surrounding injected phage DNA, we determine which proteins interact with phage proteins injected with the phage genome. Here, we surprisingly identify host proteins related to membrane and lipid biology co-purifying with the injected phage protein. Staining of infected cells with lipophilic dyes revealed a clustering of host lipids co-localized with phage DNA and protein in an early phage infection (EPI) vesicle. Early gene expression is induced by the co-injected virion RNAP (vRNAP), with specific early proteins then associating with the EPI vesicle and ribosomes, likely to facilitate localized translation. As the infection progresses, the phage genome separates from the EPI vesicle and vRNAP, moving into the nascent proteinaceous phage nucleus. Enzymes involved in DNA replication and CRISPR/restriction immune nucleases notably do not localize to the EPI vesicle, supporting that DNA is not exposed at this early time point. We therefore propose that the EPI vesicle is rapidly constructed together with injected phage proteins, phage DNA, host lipids, and host membrane proteins to enable genome protection, early transcription, localized translation, and to ensure the faithful transfer of the single copy phage genome to the proteinaceous nucleus.
]]></description>
<dc:creator>Mozumdar, D.</dc:creator>
<dc:creator>Fossati, A.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Guan, J.</dc:creator>
<dc:creator>Nieweglowska, E. S.</dc:creator>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Agard, D.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2024-03-24</dc:date>
<dc:identifier>doi:10.1101/2024.03.24.586471</dc:identifier>
<dc:title><![CDATA[Defining the constituents and functions of a lipid-based jumbo phage compartment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.22.581997v1?rss=1">
<title>
<![CDATA[
CITE-seq analysis reveals human cytomegalovirus and diabetes-associated adaptive NK cell alterations in cardiovascular disease. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.581997v1?rss=1</link>
<description><![CDATA[
Coronary artery disease (CAD) is a leading cause of mortality worldwide with Diabetes and human cyto-megalovirus (HCMV) infection as risk factors. CADs influence on human NK cells is not well characterized. CITE-seq analysis of a CAD cohort of 61 patients revealed distinctly higher NK cell SPON2 expression and lower IFNG expression in severe CAD patients. Interestingly, HCMV+ patients displayed lower SPON2 ex-pression while diabetes status reversed the HCMV effect. Diabetes led to diminished adaptive Fc{varepsilon}RI{gamma}-/low NK cell frequencies and was associated with a higher PBMC IL15/TGFB transcript ratio, while TGFB in-creased in severe CAD. SPON2 expression corresponded to changes in conventional vs. adaptive NK cell frequencies, and SPON2/IFNG ratio decreased in inflamed plaque tissue with an increased adaptive NK cell gene signature and was increased in severe CAD patients. Our results indicate that the SPON2/IFNG ra-tio and adaptive NK cell gene signature associated with stenosis severity or inflammation in CAD.
]]></description>
<dc:creator>Armstrong, S. S.</dc:creator>
<dc:creator>Chen, D. G.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Heath, J. R.</dc:creator>
<dc:creator>Feinstein, M.</dc:creator>
<dc:creator>Greenland, J. R.</dc:creator>
<dc:creator>Calabrese, D. R.</dc:creator>
<dc:creator>Lanier, L. L.</dc:creator>
<dc:creator>Ley, K.</dc:creator>
<dc:creator>Shemesh, A.</dc:creator>
<dc:date>2024-03-26</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.581997</dc:identifier>
<dc:title><![CDATA[CITE-seq analysis reveals human cytomegalovirus and diabetes-associated adaptive NK cell alterations in cardiovascular disease.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.22.584808v1?rss=1">
<title>
<![CDATA[
Screening and characterization of 133 physiologically-relevant environmental chemicals for reproductive toxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.584808v1?rss=1</link>
<description><![CDATA[
Reproduction is a functional outcome that relies on complex cellular, tissue, and organ interactions that span the developmental period to adulthood. Thus, the assessment of its disruption by environmental chemicals is remarkably painstaking in conventional toxicological animal models and does not scale up to the number of chemicals present in our environment and requiring testing.

We adapted a previously described low-throughput in vivo chromosome segregation assay using C. elegans predictive of reproductive toxicity and leveraged available public data sources (ToxCast, ICE) to screen and characterize 133 physiologically-relevant chemicals in a high-throughput manner. The screening outcome was further validated in a second, independent in vivo assay assessing embryonic viability. In total, 13 chemicals were classified as reproductive toxicants with the two most active chemicals belonging to the large family of Quaternary Ammonium Compounds (QACs) commonly used as disinfectants but with limited available reproductive toxicity data. We compared the results from the C. elegans assay with ToxCast in vitro data compiled from 700+ cell response assays and 300+ signaling pathways-based assays. We did not observe a difference in the bioactivity or in average potency (AC50) between the top and bottom chemicals. However, the intended target categories were significantly different between the classified chemicals with, in particular, an over-representation of steroid hormone targets for the high Z-score chemicals.

Taken together, these results point to the value of in vivo models that scale to high-throughput level for reproductive toxicity assessment and to the need to prioritize the assessment of QACs impacts on reproduction.
]]></description>
<dc:creator>Ulaganathan, G.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Canio, N.</dc:creator>
<dc:creator>Oke, A.</dc:creator>
<dc:creator>Armstrong, S. S.</dc:creator>
<dc:creator>Abrahamsson, D.</dc:creator>
<dc:creator>Varshavsky, J. R.</dc:creator>
<dc:creator>Lam, J.</dc:creator>
<dc:creator>Cooper, C.</dc:creator>
<dc:creator>Robinson, J. F.</dc:creator>
<dc:creator>Fung, J.</dc:creator>
<dc:creator>Woodruff, T. J.</dc:creator>
<dc:creator>Allard, P.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.584808</dc:identifier>
<dc:title><![CDATA[Screening and characterization of 133 physiologically-relevant environmental chemicals for reproductive toxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.22.584986v1?rss=1">
<title>
<![CDATA[
Broad therapeutic benefit of myosin inhibition in hypertrophic cardiomyopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.584986v1?rss=1</link>
<description><![CDATA[
Myosin inhibitor mavacamten is the only targeted treatment available for hypertrophic cardiomyopathy (HCM), a disease caused by hundreds of genetic variants that affect mainly sarcomeric myosin and its negative regulator cardiac myosin-binding protein C (cMyBP-C, encoded by MYBPC3). Here, we have examined whether the reported limited efficacy of mavacamten in a fraction of HCM patients can result from dissimilar HCM pathomechanisms triggered by different genetic variants, a scenario particularly relevant for MYBPC3-associated HCM. To this aim, we have generated knock-in mice including missense pathogenic variant cMyBP-C p.R502W, which, different from patients who carry truncations in the protein, develop progressive pathogenic myocardial remodeling in the absence of alterations of cMyBP-C levels and localization. Mechanistically, we find that mutation R502W reduces the binding affinity of cMyBP-C for myosin without inducing a shift towards more active myosin conformations as observed when cMyBP-C levels are reduced. Despite these diverging molecular alterations, we show that mavacamten blunts myocardial remodeling both in R502W and cMyBP-C-deficient, knock-out hearts. These beneficial effects are accompanied by improved tolerance to exercise only in R502W animals. Hence, our results indicate that myosin inhibition is effective to treat HCM caused by both truncating and missense variants in MYBPC3 regardless of the primary pathomechanisms they elicit.
]]></description>
<dc:creator>Sen Martin, L.</dc:creator>
<dc:creator>Fernandez Trasancos, A.</dc:creator>
<dc:creator>Lopez Unzu, M. A.</dc:creator>
<dc:creator>Pathak, D.</dc:creator>
<dc:creator>Ferrarini, A.</dc:creator>
<dc:creator>Labrador Cantarero, V.</dc:creator>
<dc:creator>Sanchez Ortiz, D.</dc:creator>
<dc:creator>Pricolo, M. R.</dc:creator>
<dc:creator>Vicente, N.</dc:creator>
<dc:creator>Velazquez Carreras, D.</dc:creator>
<dc:creator>Sanchez Garcia, L.</dc:creator>
<dc:creator>Nicolas Avila, J. A.</dc:creator>
<dc:creator>Sanchez Diaz, M.</dc:creator>
<dc:creator>Schlossarek, S.</dc:creator>
<dc:creator>Cusso, L.</dc:creator>
<dc:creator>Desco, M.</dc:creator>
<dc:creator>Villalba Orero, M.</dc:creator>
<dc:creator>Guzman Martinez, G.</dc:creator>
<dc:creator>Calvo, E.</dc:creator>
<dc:creator>Barriales Villa, R.</dc:creator>
<dc:creator>Vazquez, J.</dc:creator>
<dc:creator>Sanchez Cabo, F.</dc:creator>
<dc:creator>Hidalgo, A.</dc:creator>
<dc:creator>Carrier, L.</dc:creator>
<dc:creator>Spudich, J.</dc:creator>
<dc:creator>Ruppel, K. M.</dc:creator>
<dc:creator>Alegre Cebollada, J.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.584986</dc:identifier>
<dc:title><![CDATA[Broad therapeutic benefit of myosin inhibition in hypertrophic cardiomyopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.22.586305v1?rss=1">
<title>
<![CDATA[
Spectral graph model for fMRI: a biophysical, connectivity-based generative model for the analysis of frequency-resolved resting state fMRI 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.586305v1?rss=1</link>
<description><![CDATA[
Resting state functional MRI (rs-fMRI) is a popular and widely used technique to explore the brains functional organization and to examine if it is altered in neurological or mental disorders. The most common approach for its analysis targets the measurement of the synchronized fluctuations between brain regions, characterized as functional connectivity (FC), typically relying on pairwise correlations in activity across different brain regions. While hugely successful in exploring state- and disease-dependent network alterations, these statistical graph theory tools suffer from two key limitations. First, they discard useful information about the rich frequency content of the fMRI signal. The rich spectral information now achievable from advances in fast multiband acquisitions is consequently being under-utilized. Second, the analyzed FCs are phenomenological without a direct neurobiological underpinning in the underlying structures and processes in the brain. There does not currently exist a complete generative model framework for whole brain resting fMRI that is informed by its underlying biological basis in the structural connectome. Here we propose that a different approach can solve both challenges at once: the use of an appropriately realistic yet parsimonious biophysical signal generation model followed by graph spectral (i.e. eigen) decomposition. We call this model a Spectral Graph Model (SGM) for fMRI, using which we can not only quantify the structure-function relationship in individual subjects, but also condense the variable and individual-specific repertoire of fMRI signals spectral and spatial features into a small number of biophysically-interpretable parameters. We expect this model-based inference of rs-fMRI that seamlessly integrates with structure can be used to examine state and trait characteristics of structure-function relations in a variety of brain disorders.
]]></description>
<dc:creator>Raj, A.</dc:creator>
<dc:creator>Sipes, B. S.</dc:creator>
<dc:creator>Verma, P.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>Biswal, B. B.</dc:creator>
<dc:creator>Nagarajan, S. S.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.586305</dc:identifier>
<dc:title><![CDATA[Spectral graph model for fMRI: a biophysical, connectivity-based generative model for the analysis of frequency-resolved resting state fMRI]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.25.586565v1?rss=1">
<title>
<![CDATA[
Mixed clonal-aggregative multicellularity entrained by extreme salinity fluctuations in a close relative of animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.25.586565v1?rss=1</link>
<description><![CDATA[
Multicellularity evolved multiple times independently during eukaryotic diversification1-4. Two distinct mechanisms underpin multicellularity5: clonality (serial cell division without sister-cell separation) and aggregation (whereby independent cells assemble into a multicellular entity). Clonal and aggregative multicellularity are traditionally considered mutually exclusive1,6-9, with rare exceptions10, and evolutionary hypotheses have addressed why multicellularity might diverge toward one or the other extreme3,4. Both animals and their sister group, the choanoflagellates, are currently only known to acquire multicellularity clonally4,11-13. Here, we show that the choanoflagellate Choanoeca flexa14 forms motile and contractile cell monolayers (or "sheets") through multiple mechanisms: C. flexa sheets can form purely clonally, purely aggregatively, or by a combination of both processes. We characterise the life history of C. flexa in its natural environment - ephemeral splash pools on the island of Curacao - and show that C. flexa undergoes reversible transitions between unicellularity and multicellularity during cycles of evaporation and refilling. Different splash pools house genetically distinct strains of C. flexa, between which aggregation is constrained by kin recognition15-18. We show that clonal-aggregative multicellularity serves as a versatile strategy for the robust re-establishment of multicellularity in this variable and fast-fluctuating environment. Our findings challenge former generalisations about choanoflagellates and expand the option space of choanozoan multicellularity.
]]></description>
<dc:creator>Ros-Rocher, N.</dc:creator>
<dc:creator>Reyes-Rivera, J.</dc:creator>
<dc:creator>Foroughijabbari, Y.</dc:creator>
<dc:creator>Combredet, C.</dc:creator>
<dc:creator>Larson, B. T.</dc:creator>
<dc:creator>Coyle, M. C.</dc:creator>
<dc:creator>Houtepen, E. A. T.</dc:creator>
<dc:creator>Vermeij, M. J. A.</dc:creator>
<dc:creator>King, N.</dc:creator>
<dc:creator>Brunet, T.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.25.586565</dc:identifier>
<dc:title><![CDATA[Mixed clonal-aggregative multicellularity entrained by extreme salinity fluctuations in a close relative of animals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.27.587039v1?rss=1">
<title>
<![CDATA[
Transcript-specific enrichment enables profiling rare cell states via scRNA-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.27.587039v1?rss=1</link>
<description><![CDATA[
Single-cell genomics technologies have accelerated our understanding of cell-state heterogeneity in diverse contexts. Although single-cell RNA sequencing (scRNA-seq) identifies many rare populations of interest that express specific marker transcript combinations, traditional flow sorting limits our ability to enrich these populations for further profiling, including requiring cell surface markers with high-fidelity antibodies. Additionally, many single-cell studies require the isolation of nuclei from tissue, eliminating the ability to enrich learned rare cell states based on extranuclear protein markers. To address these limitations, we describe Programmable Enrichment via RNA Flow-FISH by sequencing (PERFF-seq), a scalable assay that enables scRNA-seq profiling of subpopulations from complex cellular mixtures defined by the presence or absence of specific RNA transcripts. Across immune populations (n = 141,227 cells) and fresh-frozen and formalin-fixed paraffin-embedded brain tissue (n = 29,522 nuclei), we demonstrate the sorting logic that can be used to enrich for cell populations via RNA-based cytometry followed by high-throughput scRNA-seq. Our approach provides a rational, programmable method for studying rare populations identified by one or more marker transcripts.
]]></description>
<dc:creator>Abay, T.</dc:creator>
<dc:creator>Stickels, R. R.</dc:creator>
<dc:creator>Takizawa, M. T.</dc:creator>
<dc:creator>Nalbant, B. N.</dc:creator>
<dc:creator>Hsieh, Y.-H.</dc:creator>
<dc:creator>Hwang, S.</dc:creator>
<dc:creator>Snopkowski, C.</dc:creator>
<dc:creator>Yu, K. K. H.</dc:creator>
<dc:creator>Abou-Mrad, Z.</dc:creator>
<dc:creator>Tabar, V.</dc:creator>
<dc:creator>Ludwig, L. S.</dc:creator>
<dc:creator>Chaligne, R.</dc:creator>
<dc:creator>Satpathy, A. T.</dc:creator>
<dc:creator>Lareau, C. A.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.587039</dc:identifier>
<dc:title><![CDATA[Transcript-specific enrichment enables profiling rare cell states via scRNA-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.26.586706v1?rss=1">
<title>
<![CDATA[
Stable and Oscillatory Hypoxia Differentially Regulate Invasibility of Breast Cancer Associated Fibroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.26.586706v1?rss=1</link>
<description><![CDATA[
As local regions in the tumor outstrip their oxygen supply, hypoxia can develop, affecting not only the cancer cells, but also other cells in the microenvironment, including cancer associated fibroblasts (CAFs). Hypoxia is also not necessarily stable over time, and can fluctuate or oscillate. Hypoxia Inducible Factor-1 is the master regulator of cellular response to hypoxia, and can also exhibit oscillations in its activity. To understand how stable, and fluctuating hypoxia influence breast CAFs, we measured changes in gene expression in CAFs in normoxia, hypoxia, and oscillatory hypoxia, as well as measured change in their capacity to resist, or assist breast cancer invasion. We show that hypoxia has a profound effect on breast CAFs causing activation of key pathways associated with fibroblast activation, but reduce myofibroblast activation and traction force generation. We also found that oscillatory hypoxia, while expectedly resulted in a "sub-hypoxic" response in gene expression, it resulted in specific activation of pathways associated with actin polymerization and actomyosin maturation. Using traction force microscopy, and a nanopatterned stromal invasion assay, we show that oscillatory hypoxia increases contractile force generation vs stable hypoxia, and increases heterogeneity in force generation response, while also additively enhancing invasibility of CAFs to MDA-MB-231 invasion. Our data show that stable and unstable hypoxia can regulate many mechnobiological characteristics of CAFs, and can contribute to transformation of CAFs to assist cancer dissemination and onset of metastasis.
]]></description>
<dc:creator>Du, W.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Afzal, J.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Suhail, Y.</dc:creator>
<dc:creator>Kshitiz,</dc:creator>
<dc:date>2024-03-28</dc:date>
<dc:identifier>doi:10.1101/2024.03.26.586706</dc:identifier>
<dc:title><![CDATA[Stable and Oscillatory Hypoxia Differentially Regulate Invasibility of Breast Cancer Associated Fibroblasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.27.586873v1?rss=1">
<title>
<![CDATA[
Conserved jumbo phage factors required for protein import into a phage nucleus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.27.586873v1?rss=1</link>
<description><![CDATA[
Bacteriophages use diverse mechanisms to evade anti-phage defenses systems. {Phi}KZ-like jumbo phages assemble a proteinaceous nucleus-like compartment that excludes antagonistic host nucleases, while internalizing DNA replication and transcription machinery1,2,3,4. The phage factors required for protein import and the mechanisms of selectivity remain unknown, however. Here, we uncover an import system composed of proteins highly conserved across nucleus-forming phages, together with additional cargo-specific contributors. Using a genetic selection that forces the phage to decrease or abolish import of specific proteins, we determine that the import of five different phage nuclear-localized proteins (Nlp) all require distinct interfaces of the same factor, Imp1 (gp69). Imp1 forms discrete puncta in the phage nuclear periphery likely in complex with a direct interactor Imp6 (gp67), a conserved protein encoded nearby. The import of some proteins, including a host topoisomerase (TopA), additionally require Imp3 (gp59), a factor required for proper Imp1 function. Three additional phage proteins (Imp2, Imp4, Imp5) are also required for the import of two queried nuclear cargos, perhaps acting as specific adaptors. We therefore propose a core import system including Imp1, Imp3, and Imp6 with the highly selective Imp1 protein licensing transport through a protein lattice.
]]></description>
<dc:creator>Kokontis, C.</dc:creator>
<dc:creator>Klein, T. A.</dc:creator>
<dc:creator>Silas, S.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2024-03-28</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.586873</dc:identifier>
<dc:title><![CDATA[Conserved jumbo phage factors required for protein import into a phage nucleus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.27.587068v1?rss=1">
<title>
<![CDATA[
The Molecular Architecture of the Nuclear Basket 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.27.587068v1?rss=1</link>
<description><![CDATA[
The nuclear pore complex (NPC) is the sole mediator of nucle-ocytoplasmic transport. Despite great advances in understanding its conserved core architecture, the peripheral regions can exhibit considerable variation within and between species. One such structure is the cage-like nuclear basket. Despite its crucial roles in mRNA surveillance and chromatin organization, an architectural understanding has remained elusive. Using in-cell cryo-electron tomography and subtomogram analysis, we explored the NPCs structural variations and the nuclear basket across fungi (yeast; S. cerevisiae), mammals (mouse; M. musculus), and protozoa (T. gondii). Using integrative structural modeling, we computed a model of the basket in yeast and mammals that revealed how a hub of Nups in the nuclear ring binds to basket-forming Mlp/Tpr proteins: the coiled-coil domains of Mlp/Tpr form the struts of the basket, while their unstructured termini constitute the basket distal densities, which potentially serve as a docking site for mRNA preprocessing before nucleocytoplasmic transport
]]></description>
<dc:creator>Singh, D.</dc:creator>
<dc:creator>Soni, N.</dc:creator>
<dc:creator>Hutchings, J.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Shaikh, F.</dc:creator>
<dc:creator>Duquette, M.</dc:creator>
<dc:creator>Suslov, S.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>van Eeuwen, T.</dc:creator>
<dc:creator>Molloy, K.</dc:creator>
<dc:creator>Shi, Y.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Guo, Q.</dc:creator>
<dc:creator>Chait, B. T.</dc:creator>
<dc:creator>Fernandez-Martinez, J.</dc:creator>
<dc:creator>Rout, M. P.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Villa, E.</dc:creator>
<dc:date>2024-03-28</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.587068</dc:identifier>
<dc:title><![CDATA[The Molecular Architecture of the Nuclear Basket]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.25.586501v1?rss=1">
<title>
<![CDATA[
py_ped_sim - A flexible forward genetic simulator for complex family pedigree analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.25.586501v1?rss=1</link>
<description><![CDATA[
BackgroundLarge-scale family pedigrees are commonly used across medical, evolutionary, and forensic genetics. These pedigrees are tools for identifying genetic disorders, tracking evolutionary patterns, and establishing familial relationships via forensic genetic identification. However, there is a lack of software to accurately simulate different pedigree structures along with genomes corresponding to those individuals in a family pedigree. This limits simulation-based evaluations of methods that use pedigrees.

ResultsWe have developed a python command-line-based tool called py_ped_sim that facilitates the simulation of pedigree structures and the genomes of individuals in a pedigree. py_ped_sim represents pedigrees as directed acyclic graphs, enabling conversion between standard pedigree formats and integration with the forward population genetic simulator, SLiM. Notably, py_ped_sim allows the simulation of varying numbers of offspring for a set of parents, with the capacity to shift the distribution of sibship sizes over generations. We additionally add simulations for events of misattributed paternity, which offers a way to simulate half-sibling relationships. We validated the accuracy of our software by simulating genomes onto diverse family pedigree structures, showing that the estimated kinship coefficients closely approximated expected values.

Conclusionspy_ped_sim is a user-friendly and open-source solution for simulating pedigree structures and conducting pedigree genome simulations. It empowers medical, forensic, and evolutionary genetics researchers to gain deeper insights into the dynamics of genetic inheritance and relatedness within families.
]]></description>
<dc:creator>Guardado, M. A.</dc:creator>
<dc:creator>Perez, C.</dc:creator>
<dc:creator>Campana, S.</dc:creator>
<dc:creator>Chavez-Rojas, B.</dc:creator>
<dc:creator>Magana, J.</dc:creator>
<dc:creator>Jackson, S.</dc:creator>
<dc:creator>Samperio, E.</dc:creator>
<dc:creator>Syas, K.</dc:creator>
<dc:creator>Hernandez, S.</dc:creator>
<dc:creator>Hernandez, R.</dc:creator>
<dc:creator>Zavala, E.</dc:creator>
<dc:creator>Rohlfs, R.</dc:creator>
<dc:date>2024-03-29</dc:date>
<dc:identifier>doi:10.1101/2024.03.25.586501</dc:identifier>
<dc:title><![CDATA[py_ped_sim - A flexible forward genetic simulator for complex family pedigree analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.25.586619v1?rss=1">
<title>
<![CDATA[
Challenges and Efficacy of Astrocyte-to-Neuron Reprogramming in Spinal Cord Injury: In Vitro Insights and In Vivo Outcomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.25.586619v1?rss=1</link>
<description><![CDATA[
Traumatic spinal cord injury (SCI) leads to the disruption of neural pathways, causing loss of neural cells, with subsequent reactive gliosis and tissue scarring that limit endogenous repair. One potential therapeutic strategy to address this is to target reactive scar-forming astrocytes with direct cellular reprogramming to convert them into neurons, by overexpression of neurogenic transcription factors. Here we used lentiviral constructs to overexpress Ascl1 or a combination of microRNAs (miRs) miR124, miR9/9*and NeuroD1 transfected into cultured and in vivo astrocytes. In vitro experiments revealed cortically-derived astrocytes display a higher efficiency (70%) of reprogramming to neurons than spinal cord-derived astrocytes. In a rat cervical SCI model, the same strategy induced only limited reprogramming of astrocytes. Delivery of reprogramming factors did not significantly affect patterns of breathing under baseline and hypoxic conditions, but significant differences in average diaphragm amplitude were seen in the reprogrammed groups during eupneic breathing, hypoxic, and hypercapnic challenges. These results show that while cellular reprogramming can be readily achieved in carefully controlled in vitro conditions, achieving a similar degree of successful reprogramming in vivo is challenging and may require additional steps.
]]></description>
<dc:creator>NICEFORO, A.</dc:creator>
<dc:creator>Zholudeva, L. V.</dc:creator>
<dc:creator>Fernandes, S.</dc:creator>
<dc:creator>Lane, M. A.</dc:creator>
<dc:creator>Qiang, L.</dc:creator>
<dc:date>2024-03-29</dc:date>
<dc:identifier>doi:10.1101/2024.03.25.586619</dc:identifier>
<dc:title><![CDATA[Challenges and Efficacy of Astrocyte-to-Neuron Reprogramming in Spinal Cord Injury: In Vitro Insights and In Vivo Outcomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.25.586698v1?rss=1">
<title>
<![CDATA[
COCONUT: An analysis of coiled-coil regions in proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.25.586698v1?rss=1</link>
<description><![CDATA[
MotivationThe molecular rules determine the strength and orientation (parallel or antiparallel) of interacting coiled-coil helices in protein-protein interactions. Interpreting these rules is crucial for identifying novel protein-protein interactions, designing competitive binders, and constructing large assemblies containing coiled-coil domains. This study establishes the molecular principles that dictate the strength and orientation of coiled-coil interactions, providing insights relevant to these applications.

ResultsWe examined how hydrophobic contacts determine structural specificity within coiled-coil dimers. Our analysis revealed that the hydrophobic core densities differ between parallel and antiparallel dimer confirmations, highlighting their importance in stabilizing different structural arrangements. We developed COiled-COil aNalysis UTility (COCONUT), a computational platform with machine learning models, validated for predictive capabilities in various applications. Using COCONUTs pipeline for coiled-coil analysis and modeling, we predicted the orientation of substitution-sensitive coiled-coil dimer, identified residue pairings in non-canonical coiled-coil heterodimer, and constructed n-stranded coiled-coil model. These results demonstrate COCONUTs utility as a computational framework for interpreting and modeling coiled-coil structures.

Availability and implementationCOCONUT is an open-source and free Python package available here https://github.com/neeleshsoni21/COCONUT. The documentation is available in the source code and here: https://neeleshsoni21.github.io/COCONUT/
]]></description>
<dc:creator>Soni, N.</dc:creator>
<dc:creator>Madhusudhan, M. S.</dc:creator>
<dc:date>2024-03-29</dc:date>
<dc:identifier>doi:10.1101/2024.03.25.586698</dc:identifier>
<dc:title><![CDATA[COCONUT: An analysis of coiled-coil regions in proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.28.587249v1?rss=1">
<title>
<![CDATA[
Plasma Proteomics of Genetic Brain Arteriosclerosis and Dementia Syndrome Identifies Signatures of Fibrosis, Angiogenesis, and Metabolic Alterations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.28.587249v1?rss=1</link>
<description><![CDATA[
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common monogenic form of vascular cognitive impairment and dementia. A genetic arteriolosclerotic disease, the molecular mechanisms driving vascular brain degeneration and decline remain unclear. With the goal of driving discovery of disease-relevant biological perturbations in CADASIL, we used machine learning approaches to extract proteomic disease signatures from large-scale proteomics generated from plasma collected from three distinct cohorts in US and Colombia: CADASIL-Early (N = 53), CADASIL-Late (N = 45), and CADASIL-Colombia (N = 71). We extracted molecular signatures with high predictive value for early and late-stage CADASIL and performed robust cross- and external-validation. We examined the biological and clinical relevance of our findings through pathway enrichment analysis and testing of associations with clinical outcomes. Our study represents a model for unbiased discovery of molecular signatures and disease biomarkers, combining non-invasive plasma proteomics with clinical data. We report on novel disease-associated molecular signatures for CADASIL, derived from the accessible plasma proteome, with relevance to vascular cognitive impairment and dementia.
]]></description>
<dc:creator>Keller, J. N.</dc:creator>
<dc:creator>Radabaugh, H.</dc:creator>
<dc:creator>Karvelas, N.</dc:creator>
<dc:creator>Fitzsimons, S.</dc:creator>
<dc:creator>Treiman, S.</dc:creator>
<dc:creator>Palafox, M. F.</dc:creator>
<dc:creator>McDonnell, L.</dc:creator>
<dc:creator>Quiroz, Y. T.</dc:creator>
<dc:creator>Lopera, F. J.</dc:creator>
<dc:creator>Banerjee, D.</dc:creator>
<dc:creator>Wang, M. M.</dc:creator>
<dc:creator>Arboleda-Velasquez, J. F.</dc:creator>
<dc:creator>Meschia, J. F.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Elahi, F. M.</dc:creator>
<dc:date>2024-03-29</dc:date>
<dc:identifier>doi:10.1101/2024.03.28.587249</dc:identifier>
<dc:title><![CDATA[Plasma Proteomics of Genetic Brain Arteriosclerosis and Dementia Syndrome Identifies Signatures of Fibrosis, Angiogenesis, and Metabolic Alterations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.27.587047v1?rss=1">
<title>
<![CDATA[
Epigenomic landscape of the human dorsal root ganglion: sex differences and transcriptional regulation of nociceptive genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.27.587047v1?rss=1</link>
<description><![CDATA[
Gene expression is influenced by chromatin architecture via controlled access of regulatory factors to DNA. To better understand gene regulation in the human dorsal root ganglion (hDRG) we used bulk and spatial transposase-accessible chromatin technology followed by sequencing (ATAC-seq). Using bulk ATAC-seq, we detected that in females diverse differentially accessible chromatin regions (DARs) mapped to the X chromosome and in males to autosomal genes. EGR1/3 and SP1/4 transcription factor binding motifs were abundant within DARs in females, and JUN, FOS and other AP-1 factors in males. To dissect the open chromatin profile in hDRG neurons, we used spatial ATAC-seq. The neuron cluster showed higher chromatin accessibility in GABAergic, glutamatergic, and interferon-related genes in females, and in Ca2+-signaling-related genes in males. Sex differences in transcription factor binding sites in neuron-proximal barcodes were consistent with the trends observed in bulk ATAC-seq data. We validated that EGR1 expression is biased to female hDRG compared to male. Strikingly, XIST, the long-noncoding RNA responsible for X inactivation, hybridization signal was found to be highly dispersed in the female neuronal but not non-neuronal nuclei suggesting weak X inactivation in female hDRG neurons. Our findings point to baseline epigenomic sex differences in the hDRG that likely underlie divergent transcriptional responses that determine mechanistic sex differences in pain.
]]></description>
<dc:creator>Franco-Enzastiga, U.</dc:creator>
<dc:creator>Inturi, N. N.</dc:creator>
<dc:creator>Natarajan, K.</dc:creator>
<dc:creator>Mwirigi, J. M.</dc:creator>
<dc:creator>Mazhar, K.</dc:creator>
<dc:creator>Schlachetzki, J. C.</dc:creator>
<dc:creator>Schumacher, M.</dc:creator>
<dc:creator>Price, T. J.</dc:creator>
<dc:date>2024-03-30</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.587047</dc:identifier>
<dc:title><![CDATA[Epigenomic landscape of the human dorsal root ganglion: sex differences and transcriptional regulation of nociceptive genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.27.587093v1?rss=1">
<title>
<![CDATA[
A high-throughput method for quantifying Drosophila fecundity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.27.587093v1?rss=1</link>
<description><![CDATA[
Measurements of Drosophila fecundity are used in a wide variety of studies, such as investigations of stem cell biology, nutrition, behavior, and toxicology. In addition, because fecundity assays are performed on live flies, they are suitable for longitudinal studies such as investigations of aging or prolonged chemical exposure. However, standard Drosophila fecundity assays have been difficult to perform in a high-throughput manner because experimental factors such as the physiological state of the flies and environmental cues must be carefully controlled to achieve consistent results. In addition, exposing flies to a large number of different experimental conditions (such as chemical additives in the diet) and manually counting the number of eggs laid to determine the impact on fecundity is time-consuming. We have overcome these challenges by combining a new multiwell fly culture strategy with a novel 3D-printed fly transfer device to rapidly and accurately transfer flies from one plate to another; the RoboCam, a low-cost, custom built robotic camera to capture images of the wells automatically; and an image segmentation pipeline to automatically identify and quantify eggs. We show that this method is compatible with robust and consistent egg laying throughout the assay period; and demonstrate that the automated pipeline for quantifying fecundity is very accurate (r2 = 0.98 for the correlation between the automated egg counts and the ground truth) In addition, we show that this method can be used to efficiently detect the effects on fecundity induced by dietary exposure to chemicals. Taken together, this strategy substantially increases the efficiency and reproducibility of high throughput egg laying assays that require exposing flies to multiple different media conditions.
]]></description>
<dc:creator>Gomez, A.</dc:creator>
<dc:creator>Gonzalez, S.</dc:creator>
<dc:creator>Oke, A.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Duong, J. B.</dc:creator>
<dc:creator>Esquerra, R. M.</dc:creator>
<dc:creator>Zimmerman, T.</dc:creator>
<dc:creator>Capponi, S.</dc:creator>
<dc:creator>Fung, J. C.</dc:creator>
<dc:creator>Nystul, T. G.</dc:creator>
<dc:date>2024-03-30</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.587093</dc:identifier>
<dc:title><![CDATA[A high-throughput method for quantifying Drosophila fecundity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.27.587103v1?rss=1">
<title>
<![CDATA[
Mesolimbic dopamine ramps reflect environmental timescales 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.27.587103v1?rss=1</link>
<description><![CDATA[
Mesolimbic dopamine activity occasionally exhibits ramping dynamics, reigniting debate on theories of dopamine signaling. This debate is ongoing partly because the experimental conditions under which dopamine ramps emerge remain poorly understood. Here, we show that during Pavlovian and instrumental conditioning, mesolimbic dopamine ramps are only observed when the inter-trial interval is short relative to the trial period. These results constrain theories of dopamine signaling and identify a critical variable determining the emergence of dopamine ramps.
]]></description>
<dc:creator>Floeder, J. R.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Mohebi, A.</dc:creator>
<dc:creator>K Namboodiri, V. M.</dc:creator>
<dc:date>2024-03-30</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.587103</dc:identifier>
<dc:title><![CDATA[Mesolimbic dopamine ramps reflect environmental timescales]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.01.587588v1?rss=1">
<title>
<![CDATA[
Differential Roles of Kinetic On- and Off-Rates in T-Cell Receptor Signal Integration Revealed with a Modified Fab'-DNA Ligand 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.01.587588v1?rss=1</link>
<description><![CDATA[
Antibody-derived T-cell receptor (TCR) agonists are commonly used to activate T cells. While antibodies can trigger TCRs regardless of clonotype, they bypass native T cell signal integration mechanisms that rely on monovalent, membrane-associated, and relatively weakly-binding ligand in the context of cellular adhesion. Commonly used antibodies and their derivatives bind much more strongly than native peptide-MHC (pMHC) ligands bind their cognate TCRs. Because ligand dwell time is a critical parameter that tightly correlates with physiological function of the TCR signaling system, there is a general need, both in research and therapeutics, for universal TCR ligands with controlled kinetic binding parameters. To this end, we have introduced point mutations into recombinantly expressed -TCR{beta} H57 Fab to modulate the dwell time of monovalent Fab binding to TCR. When tethered to a supported lipid bilayer via DNA complementation, these monovalent Fab-DNA ligands activate T cells with potencies well-correlated with their TCR binding dwell time. Single-molecule tracking studies in live T cells reveal that individual binding events between Fab-DNA ligands and TCRs elicit local signaling responses closely resembling native pMHC. The unique combination of high on- and off-rate of the H57 R97L mutant enables direct observations of cooperative interplay between ligand binding and TCR-proximal condensation of the linker for activation of T cells (LAT), which is not readily visualized with pMHC. This work provides insights into how T cells integrate kinetic information from synthetic ligands and introduces a method to develop affinity panels for polyclonal T cells, such as cells from a human patient.

STATEMENT OF SIGNIFICANCET cells read kinetic information from ligands binding to T-cell receptors (TCRs) to make cell fate decisions. Unique kinetic features of a modified Fab-DNA ligand enable direct visualization multiple TCR signal coordination through a nascent LAT condensation event. We further observed positive feedback through a kinetic on-rate enhancement in the growing LAT condensate. These observations help unify several seemingly disparate aspects of TCR signaling that have been debated in the literature. Furthermore, calibration of the Fab-DNA ligand against native agonist pMHC establishes a basis for quantitative analysis of TCR signaling in polyclonal T cell populations.
]]></description>
<dc:creator>Wilhelm, K. B.</dc:creator>
<dc:creator>Vissa, A.</dc:creator>
<dc:creator>Groves, J. T.</dc:creator>
<dc:date>2024-04-02</dc:date>
<dc:identifier>doi:10.1101/2024.04.01.587588</dc:identifier>
<dc:title><![CDATA[Differential Roles of Kinetic On- and Off-Rates in T-Cell Receptor Signal Integration Revealed with a Modified Fab'-DNA Ligand]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.01.587605v1?rss=1">
<title>
<![CDATA[
RosetteArray Platform for Quantitative High-Throughput Screening of Human Neurodevelopmental Risk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.01.587605v1?rss=1</link>
<description><![CDATA[
Neural organoids have revolutionized how human neurodevelopmental disorders (NDDs) are studied. Yet, their utility for screening chemical hazards and prospective therapeutics for NDDs is limited by a lack of morphological reproducibility and cost-effective scalability. Here, we describe the RosetteArray platform, which can be used as an off-the-shelf, 96-well plate assay that standardizes incipient forebrain and spinal cord organoid morphogenesis as adherent, micropatterned, 3-D, singularly polarized neural rosette tissues ([~]200 and [~]800 per well, respectively). Seeded directly from cryopreserved human pluripotent stem cells, RosetteArrays are cultured over 6-8 days and fixed, immunostained and imaged in situ to enable artificial intelligence-based quantitative analysis. By screening the inception of [~]75,000 neural organoids throughout this manuscript, we provide proof-of-concept demonstrations of the platforms utility for detecting developmental neurotoxicity hazard and screening genetic and environmental factors known to cause clinical Neural Tube Defect risk. Given the documented perturbation of rosette morphogenesis in neural organoid models of several NDDs, the RosetteArray platform could enable quantitative high-throughput screening (qHTS) of human neurodevelopmental risk across regulatory and precision medicine applications.
]]></description>
<dc:creator>Lundin, B. F.</dc:creator>
<dc:creator>Knight, G. T.</dc:creator>
<dc:creator>Fedorchak, N. J.</dc:creator>
<dc:creator>Krucki, K.</dc:creator>
<dc:creator>Iyer, N.</dc:creator>
<dc:creator>Maher, J. E.</dc:creator>
<dc:creator>Izban, N. R.</dc:creator>
<dc:creator>Roberts, A.</dc:creator>
<dc:creator>Cicero, M. R.</dc:creator>
<dc:creator>Robinson, J. F.</dc:creator>
<dc:creator>Iskandar, B. J.</dc:creator>
<dc:creator>Willett, R.</dc:creator>
<dc:creator>Ashton, R. S.</dc:creator>
<dc:date>2024-04-02</dc:date>
<dc:identifier>doi:10.1101/2024.04.01.587605</dc:identifier>
<dc:title><![CDATA[RosetteArray Platform for Quantitative High-Throughput Screening of Human Neurodevelopmental Risk]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.02.587507v1?rss=1">
<title>
<![CDATA[
Cryo-EM Structures Reveal Tau Filaments from Down Syndrome Adopt Alzheimer's Disease Fold 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.02.587507v1?rss=1</link>
<description><![CDATA[
Down syndrome (DS) is a common genetic condition caused by trisomy of chromosome 21. Among their complex clinical features, including musculoskeletal, neurological, and cardiovascular disabilities, individuals with DS have an increased risk of developing progressive dementia and early-onset Alzheimers disease (AD). This dementia is attributed to the increased gene dosage of the amyloid-{beta} (A{beta}) precursor protein gene, the formation of self-propagating A{beta} and tau prion conformers, and the deposition of neurotoxic A{beta} plaques and tau neurofibrillary tangles. Tau amyloid fibrils have previously been established to adopt many distinct conformations across different neurodegenerative conditions. Here, we report the characterization of brain samples from four DS cases spanning 36 to 63 years of age by spectral confocal imaging with conformation-specific dyes and cryo-electron microscopy (cryo-EM) to determine structures of isolated tau fibrils. High-resolution structures revealed paired helical filament (PHF) and straight filament (SF) conformations of tau that were identical to those determined from AD cases. The PHFs and SFs are made of two C-shaped protofilaments, each containing a cross-{beta}/{beta}-helix motif. Similar to filaments from AD cases, most filaments from the DS cases adopted the PHF form, while a minority (approximately 20%) formed SFs. Samples from the youngest individual with no documented dementia had sparse tau deposits. To isolate tau for cryo-EM from this challenging sample we used a novel affinity-grid method involving a graphene oxide surface derivatized with anti-tau antibodies. This method improved isolation and revealed that primarily tau PHFs and a minor population of chronic traumatic encephalopathy type II-like filaments were present in this youngest case. These findings expand the similarities between AD and DS to the molecular level, providing insight into their related pathologies and the potential for targeting common tau filament folds by small-molecule therapeutics and diagnostics.
]]></description>
<dc:creator>Ghosh, U.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Shi, M.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Merz, G. E.</dc:creator>
<dc:creator>Prusiner, S. B.</dc:creator>
<dc:creator>Southworth, D. R.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:date>2024-04-03</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.587507</dc:identifier>
<dc:title><![CDATA[Cryo-EM Structures Reveal Tau Filaments from Down Syndrome Adopt Alzheimer's Disease Fold]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.02.587608v1?rss=1">
<title>
<![CDATA[
IRF8-driven reprogramming of the immune microenvironment enhances anti-tumor adaptive immunity and reduces immunosuppression in murine glioblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.02.587608v1?rss=1</link>
<description><![CDATA[
BackgroundGlioblastoma (GBM) has a highly immunosuppressive tumor immune microenvironment (TIME), largely mediated by myeloid-derived suppressor cells (MDSCs). Here, we utilized a retroviral replicating vector (RRV) to deliver Interferon Regulatory Factor 8 (IRF8), a master regulator of type 1 conventional dendritic cell (cDC1) development, in a syngeneic murine GBM model. We hypothesized that RRV-mediated delivery of IRF8 could "reprogram" intratumoral MDSCs into antigen-presenting cells (APCs) and thereby restore T-cell responses.

MethodsEffects of RRV-IRF8 on survival and tumor growth kinetics were examined in the SB28 murine GBM model. Immunophenotype was analyzed by flow cytometry and gene expression assays. We assayed functional immunosuppression and antigen presentation by ex vivo T-cell-myeloid co-culture.

ResultsMice with RRV-IRF8 pre-transduced intracerebral tumors had significantly longer survival and slower tumor growth compared to controls. RRV-IRF8 treated tumors exhibited significant enrichment of cDC1s and CD8+ T-cells. Additionally, myeloid cells derived from RRV-IRF8 tumors showed decreased expression of the immunosuppressive markers Arg1 and IDO1 and demonstrated reduced suppression of naive T-cell proliferation in ex vivo co-culture, compared to controls. Furthermore, DCs from RRV-IRF8 tumors showed increased antigen presentation compared to those from control tumors. In vivo treatment with azidothymidine (AZT), a viral replication inhibitor, showed that IRF8 transduction in both tumor and non-tumor cells is necessary for survival benefit, associated with a reprogrammed, cDC1- and CD8 T-cell-enriched TIME.

ConclusionsOur results indicate that reprogramming of glioma-infiltrating myeloid cells by in vivo expression of IRF8 may reduce immunosuppression and enhance antigen presentation, achieving improved tumor control.

Key pointsO_LIGBM intra-tumoral myeloid cells are proliferative and targets for RRV therapy.
C_LIO_LIExpression of IRF8 significantly improves survival and slows tumor growth in murine GBM.
C_LIO_LIIRF8 expression in MDSCs reduces immunosuppression and enriches cDC1s in vivo.
C_LI

Importance of the studyRecent publications have presented conflicting studies regarding the role of IRF8 in GBM. While some studies showed IRF8 as a negative prognostic factor, others demonstrated the conversion of tumor cells into DCs using IRF8. Here, we show that RRV-mediated delivery of IRF8, a clinically relevant modality, allows for transduction of both tumor and immune cells in vivo. We show that a significant survival effect relies heavily on the infection and modulation of both populations, and that even a modest number of reprogrammed intra-tumoral MDSCs can have a substantial impact on the immunological milieu, significantly enriching and activating cytotoxic T-cells. Further, this work reveals intra-tumoral myeloid cells as a target for other RRV-based gene therapies.
]]></description>
<dc:creator>Montoya, M.</dc:creator>
<dc:creator>Collins, S. A.</dc:creator>
<dc:creator>Chuntova, P.</dc:creator>
<dc:creator>Patel, T. S.</dc:creator>
<dc:creator>Nejo, T.</dc:creator>
<dc:creator>Yamamichi, A.</dc:creator>
<dc:creator>Kasahara, N.</dc:creator>
<dc:creator>Okada, H.</dc:creator>
<dc:date>2024-04-03</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.587608</dc:identifier>
<dc:title><![CDATA[IRF8-driven reprogramming of the immune microenvironment enhances anti-tumor adaptive immunity and reduces immunosuppression in murine glioblastoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.02.587832v1?rss=1">
<title>
<![CDATA[
Optineurin-facilitated axonal mitochondria delivery promotes neuroprotection and axon regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.02.587832v1?rss=1</link>
<description><![CDATA[
Optineurin (OPTN) mutations are linked to amyotrophic lateral sclerosis (ALS) and normal tension glaucoma (NTG), but a relevant animal model is lacking, and the molecular mechanisms underlying neurodegeneration are unknown. We found that OPTN C-terminus truncation (OPTN{Delta}C) causes late-onset neurodegeneration of retinal ganglion cells (RGCs), optic nerve (ON), and spinal cord motor neurons, preceded by a striking decrease of axonal mitochondria. Surprisingly, we discover that OPTN directly interacts with both microtubules and the mitochondrial transport complex TRAK1/KIF5B, stabilizing them for proper anterograde axonal mitochondrial transport, in a C- terminus dependent manner. Encouragingly, overexpressing OPTN/TRAK1/KIF5B reverses not only OPTN truncation-induced, but also ocular hypertension-induced neurodegeneration, and promotes striking ON regeneration. Therefore, in addition to generating new animal models for NTG and ALS, our results establish OPTN as a novel facilitator of the microtubule-dependent mitochondrial transport necessary for adequate axonal mitochondria delivery, and its loss as the likely molecular mechanism of neurodegeneration.
]]></description>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Webber, H. C.</dc:creator>
<dc:creator>Bian, F.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Prakash, M.</dc:creator>
<dc:creator>Feng, X.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>You, I.-J.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Liu, P.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Chang, C.-Y.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Shah, S. H.</dc:creator>
<dc:creator>La Torre, A.</dc:creator>
<dc:creator>Welsbie, D. S.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Duan, X.</dc:creator>
<dc:creator>Goldberg, J. L.</dc:creator>
<dc:creator>Braun, M.</dc:creator>
<dc:creator>Lansky, Z.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:date>2024-04-03</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.587832</dc:identifier>
<dc:title><![CDATA[Optineurin-facilitated axonal mitochondria delivery promotes neuroprotection and axon regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.02.587841v1?rss=1">
<title>
<![CDATA[
Structure and function of an intermediate GPCR-G protein complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.02.587841v1?rss=1</link>
<description><![CDATA[
Unraveling the signaling roles of intermediate complexes is pivotal for G protein-coupled receptor (GPCR) drug development. Despite hundreds of GPCR-G{beta}{gamma} structures, these snapshots primarily capture the fully activated end-state complex. Consequently, a comprehensive understanding of the conformational transitions during GPCR activation and the roles of intermediate GPCR-G protein complexes in signaling remain elusive. Guided by a conformational landscape profiled by 19F quantitative NMR (19F-qNMR) and Molecular Dynamics (MD) simulations, we resolved the structure of an unliganded GPCR-G protein intermediate complex by blocking its transition to the fully activated end-state complex. More importantly, we presented direct evidence that the intermediate GPCR-Gs{beta}{gamma} complex initiates a rate-limited nucleotide exchange without progressing to the fully activated end-state complex, thereby bridging a significant gap in our understanding the complexity of GPCR signaling. Understanding the roles of individual conformational states and their complexes in signaling efficacy and bias will help us to design drugs that discriminately target a disease-related conformation.
]]></description>
<dc:creator>Bi, M.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>xu, J.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:creator>Adediwrua, V. A.</dc:creator>
<dc:creator>Miao, Y.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Ye, L.</dc:creator>
<dc:date>2024-04-03</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.587841</dc:identifier>
<dc:title><![CDATA[Structure and function of an intermediate GPCR-G protein complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.05.588321v1?rss=1">
<title>
<![CDATA[
Chronic hyperactivation of midbrain dopamine neurons causes preferential dopamine neuron degeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.05.588321v1?rss=1</link>
<description><![CDATA[
Parkinsons disease (PD) is characterized by the death of substantia nigra (SNc) dopamine (DA) neurons, but the pathophysiological mechanisms that precede and drive their death remain unknown. The activity of DA neurons is likely altered in PD, but we understand little about if or how chronic changes in activity may contribute to degeneration. To address this question, we developed a chemogenetic (DREADD) mouse model to chronically increase DA neuron activity, and confirmed this increase using ex vivo electrophysiology. Chronic hyperactivation of DA neurons resulted in prolonged increases in locomotor activity during the light cycle and decreases during the dark cycle, consistent with chronic changes in DA release and circadian disturbances. We also observed early, preferential degeneration of SNc projections, recapitulating the PD hallmarks of selective vulnerability of SNc axons and the comparative resilience of ventral tegmental area axons. This was followed by eventual loss of midbrain DA neurons. Continuous DREADD activation resulted in a sustained increase in baseline calcium levels, supporting a role for increased calcium in the neurodegeneration process. Finally, spatial transcriptomics from DREADD mice examining midbrain DA neurons and striatal targets, and cross-validation with human patient samples, provided insights into potential mechanisms of hyperactivity-induced toxicity and PD. Our results thus reveal the preferential vulnerability of SNc DA neurons to increased neural activity, and support a potential role for increased neural activity in driving degeneration in PD.
]]></description>
<dc:creator>Rademacher, K. L.</dc:creator>
<dc:creator>Doric, Z.</dc:creator>
<dc:creator>Haddad, D.</dc:creator>
<dc:creator>Mamaligas, A.</dc:creator>
<dc:creator>Liao, S.-C.</dc:creator>
<dc:creator>Creed, R.</dc:creator>
<dc:creator>Kano, K.</dc:creator>
<dc:creator>Chatterton, Z.</dc:creator>
<dc:creator>Fu, Y.</dc:creator>
<dc:creator>Garcia, J. H.</dc:creator>
<dc:creator>Vance, V.</dc:creator>
<dc:creator>Sei, Y.</dc:creator>
<dc:creator>Kreitzer, A.</dc:creator>
<dc:creator>Halliday, G.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Margolis, E.</dc:creator>
<dc:creator>Nakamura, K. L.</dc:creator>
<dc:date>2024-04-10</dc:date>
<dc:identifier>doi:10.1101/2024.04.05.588321</dc:identifier>
<dc:title><![CDATA[Chronic hyperactivation of midbrain dopamine neurons causes preferential dopamine neuron degeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.10.588779v1?rss=1">
<title>
<![CDATA[
Accurate estimation of intraspecific microbial gene content variation in metagenomic data with MIDAS v3 and StrainPGC 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.10.588779v1?rss=1</link>
<description><![CDATA[
Metagenomics has greatly expanded our understanding of the human gut microbiome by revealing a vast diversity of bacterial species within and across individuals. Even within a single species, different strains can have highly divergent gene content, affecting traits such as antibiotic resistance, metabolism, and virulence. Methods that harness metagenomic data to resolve strain-level differences in functional potential are crucial for understanding the causes and consequences of this intraspecific diversity. The enormous size of pangenome references, strain mixing within samples, and inconsistent sequencing depth present challenges for existing tools that analyze samples one at a time. To address this gap, we updated the MIDAS pangenome profiler, now released as version 3, and developed StrainPGC, an approach to strain-specific gene content estimation that combines strain tracking and correlations across multiple samples. We validate our integrated analysis using a complex synthetic community of strains from the human gut and find that StrainPGC outperforms existing approaches. Analyzing a large, publicly available metagenome collection from inflammatory bowel disease patients and healthy controls, we catalog the functional repertoires of thousands of strains across hundreds of species, capturing extensive diversity missing from reference databases. Finally, we apply StrainPGC to metagenomes from a clinical trial of fecal microbiota transplantation for the treatment of ulcerative colitis. We identify two Escherichia coli strains from two different donors that are both frequently transmitted to patients, but have notable differences in functional potential. StrainPGC and MIDAS v3 together enable precise, intraspecific pangenomic investigations using large collections of metagenomic data without microbial isolation or de novo assembly.
]]></description>
<dc:creator>Smith, B. J.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Dubinkina, V.</dc:creator>
<dc:creator>Jin, X.</dc:creator>
<dc:creator>Moltzau-Anderson, J.</dc:creator>
<dc:creator>Pollard, K.</dc:creator>
<dc:date>2024-04-10</dc:date>
<dc:identifier>doi:10.1101/2024.04.10.588779</dc:identifier>
<dc:title><![CDATA[Accurate estimation of intraspecific microbial gene content variation in metagenomic data with MIDAS v3 and StrainPGC]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.17.589938v1?rss=1">
<title>
<![CDATA[
A shift from pleiotropic to modular adaptation revealed by a high-resolution two-step adaptive walk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.17.589938v1?rss=1</link>
<description><![CDATA[
Evolution by natural selection is expected to be a slow and gradual process. In particular, the mutations that drive evolution are predicted to be small and modular, incrementally improving a small number of traits. However, adaptive mutations identified early in microbial evolution experiments, cancer, and other systems often provide substantial fitness gains and pleiotropically improve multiple traits at once. We asked whether such pleiotropically adaptive mutations are common throughout adaptation or are instead a rare feature of early steps in evolution that tend to target key signaling pathways. To do so, we conducted barcoded second-step evolution experiments initiated from five first-step mutations identified from a prior yeast evolution experiment. We then isolated hundreds of second-step mutations from these evolution experiments, measured their fitness and performance in several growth phases, and conducted whole-genome sequencing of the second-step clones. Here, we found that while the vast majority of mutants isolated from the first-step of evolution in this condition show patterns of pleiotropic adaptation - improving both performance in fermentation and respiration growth phases - second-step mutations show a shift towards modular adaptation, mostly improving respiration performance and only rarely improving fermentation performance. We also identified a shift in the molecular basis of adaptation from genes in cellular signaling pathways towards genes involved in respiration and mitochondrial function. Our results suggest that the genes in cellular signaling pathways are particularly capable of providing large, adaptively pleiotropic benefits to the organism due to their ability to coherently affect many phenotypes at once. As such, these genes may serve as the source of pleiotropic adaptation in the early stages of evolution, and once these become exhausted, organisms then adapt more gradually, acquiring smaller, more modular mutations.
]]></description>
<dc:creator>Kinsler, G.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Sherlock, G. J.</dc:creator>
<dc:creator>Petrov, D.</dc:creator>
<dc:date>2024-04-18</dc:date>
<dc:identifier>doi:10.1101/2024.04.17.589938</dc:identifier>
<dc:title><![CDATA[A shift from pleiotropic to modular adaptation revealed by a high-resolution two-step adaptive walk]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.17.590000v1?rss=1">
<title>
<![CDATA[
Molecular basis of proton-sensing by G protein-coupled receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.17.590000v1?rss=1</link>
<description><![CDATA[
Three proton-sensing G protein-coupled receptors (GPCRs), GPR4, GPR65, and GPR68, respond to changes in extracellular pH to regulate diverse physiology and are implicated in a wide range of diseases. A central challenge in determining how protons activate these receptors is identifying the set of residues that bind protons. Here, we determine structures of each receptor to understand the spatial arrangement of putative proton sensing residues in the active state. With a newly developed deep mutational scanning approach, we determined the functional importance of every residue in proton activation for GPR68 by generating [~]9,500 mutants and measuring effects on signaling and surface expression. This unbiased screen revealed that, unlike other proton-sensitive cell surface channels and receptors, no single site is critical for proton recognition in GPR68. Instead, a network of titratable residues extend from the extracellular surface to the transmembrane region and converge on canonical class A GPCR activation motifs to activate proton-sensing GPCRs. More broadly, our approach integrating structure and unbiased functional interrogation defines a new framework for understanding the rich complexity of GPCR signaling.

One-sentence summaryThe protonation networks governing activation of human pH-sensing GPCRs are uncovered by integrative cryo-EM and deep mutational scanning.
]]></description>
<dc:creator>Howard, M. K.</dc:creator>
<dc:creator>Hoppe, N.</dc:creator>
<dc:creator>Huang, X.-P.</dc:creator>
<dc:creator>Macdonald, C. B.</dc:creator>
<dc:creator>Mehrotra, E.</dc:creator>
<dc:creator>Rockefeller Grimes, P.</dc:creator>
<dc:creator>Zahm, A. M.</dc:creator>
<dc:creator>Trinidad, D. D.</dc:creator>
<dc:creator>English, J. G.</dc:creator>
<dc:creator>Coyote-Maestas, W.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:date>2024-04-18</dc:date>
<dc:identifier>doi:10.1101/2024.04.17.590000</dc:identifier>
<dc:title><![CDATA[Molecular basis of proton-sensing by G protein-coupled receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.16.589814v1?rss=1">
<title>
<![CDATA[
Massively parallel jumping assay decodes Alu retrotransposition activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.16.589814v1?rss=1</link>
<description><![CDATA[
The human genome contains millions of retrotransposons, several of which could become active due to somatic mutations having phenotypic consequences, including disease. However, it is not thoroughly understood how nucleotide changes in retrotransposons affect their jumping activity. Here, we developed a novel massively parallel jumping assay (MPJA) that can test the jumping potential of thousands of transposons en masse. We generated nucleotide variant library of selected four Alu retrotransposons containing 165,087 different haplotypes and tested them for their jumping ability using MPJA. We found 66,821 unique jumping haplotypes, allowing us to pinpoint domains and variants vital for transposition. Mapping these variants to the Alu-RNA secondary structure revealed stem-loop features that contribute to jumping potential. Combined, our work provides a novel high-throughput assay that assesses the ability of retrotransposons to jump and identifies nucleotide changes that have the potential to reactivate them in the human genome.
]]></description>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Matharu, N.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Sohota, A.</dc:creator>
<dc:creator>Deng, L.</dc:creator>
<dc:creator>Hung, Y.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Sims, J.</dc:creator>
<dc:creator>Rattanasopha, S.</dc:creator>
<dc:creator>Meyer, J.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Kircher, M.</dc:creator>
<dc:date>2024-04-19</dc:date>
<dc:identifier>doi:10.1101/2024.04.16.589814</dc:identifier>
<dc:title><![CDATA[Massively parallel jumping assay decodes Alu retrotransposition activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.16.589837v1?rss=1">
<title>
<![CDATA[
Depletion of Protein Phosphatase 1 Results in Persistent Activation of the Integrated Stress Response and Age-Associated Exacerbation of Pulmonary Veno-Occlusive Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.16.589837v1?rss=1</link>
<description><![CDATA[
Pulmonary veno-occlusive disease (PVOD) is a form of pulmonary hypertension that affects individuals across the age spectrum. PVOD is characterized by the obstruction of small pulmonary vessels, causing increased pulmonary artery (PA) pressure and leading to right ventricular heart (RV) failure. Previous research showed that the administration of Mitomycin-C (MMC) in rats mediates PVOD through the activation of the eukaryotic initiation factor 2 (eIF2) kinase PKR and the integrated stress response (ISR), resulting in the impairment of vascular endothelial junctional structure and barrier function. In this study, we reveal that older rats experience more severe pulmonary vascular remodeling and RV hypertrophy than younger rats after MMC treatment due to lower levels of protein phosphatase 1, leading to prolonged eIF2 phosphorylation and ISR activation. We demonstrate that pharmacological blocking of the PKR-ISR pathway mitigates PVOD symptoms in both age groups, suggesting targeting the PKR-ISR axis as a potential PVOD therapeutic strategy.
]]></description>
<dc:creator>Prabhakar, A.</dc:creator>
<dc:creator>Wadhwa, M.</dc:creator>
<dc:creator>Ghatpande, P.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Graham, B. B.</dc:creator>
<dc:creator>Lagna, G.</dc:creator>
<dc:creator>Hata, A.</dc:creator>
<dc:date>2024-04-20</dc:date>
<dc:identifier>doi:10.1101/2024.04.16.589837</dc:identifier>
<dc:title><![CDATA[Depletion of Protein Phosphatase 1 Results in Persistent Activation of the Integrated Stress Response and Age-Associated Exacerbation of Pulmonary Veno-Occlusive Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.20.590402v1?rss=1">
<title>
<![CDATA[
Component specific responses of the microbiomes to common chemical stressors in the human food chain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.20.590402v1?rss=1</link>
<description><![CDATA[
Along a food chain, microbiomes occur in each component and often contribute to the functioning or the health of their host or environment.  One Health emphasizes the connectivity of each components health. Chemical stress typically causes dysbiotic microbiomes, but it remains unclear whether chemical stressors consistently affect the microbiomes along food chain components. Here, we systematically challenged a model food chain, including water, sediments, soil, plants, and animals, with three chemical stresses consisting of arsenic (a toxic trace element), benzoxazinoids (an abundant bioactive plant metabolites), and terbuthylazine (an herbicide typically found along a human food chain). The analysis of 1,064 microbiome profiles for commonalities and differences in their stress responses indicated that chemical stressors decreased microbiome diversity in soil and animal, but not in the other microbiomes. In response to stress, all food chain communities strongly shifted in their composition, generally becoming compositionally more similar to each other. In addition, we observed stochastic effects in host-associated communities (plant, animal). Dysbiotic microbiomes were characterized by different sets of bacteria, which responded specifically to the three chemical stressors. Microbial co-occurrence patterns significantly shifted with either decreased (water, sediment, plant, animal) or increased (soil) network sparsity and numbers of keystone taxa following stress treatments. This suggested major re-distribution of the roles that specific taxa may have, with the community stability of plant and animal microbiomes being the most affected by chemical stresses. Overall, we observed stress- and component-specific responses to chemical stressors in microbiomes along the model food chain, which could have implications on food chain health.
]]></description>
<dc:creator>Wasimuddin, W.</dc:creator>
<dc:creator>Chiaia-Hernandez, A.</dc:creator>
<dc:creator>Terrettaz, C.</dc:creator>
<dc:creator>Thoenen, L.</dc:creator>
<dc:creator>Caggia, V.</dc:creator>
<dc:creator>Matteo, P.</dc:creator>
<dc:creator>Coll-Crespi, M.</dc:creator>
<dc:creator>Notter, M.</dc:creator>
<dc:creator>Mukherjee, M.</dc:creator>
<dc:creator>Chavez-Capilla, T.</dc:creator>
<dc:creator>Ronchi, F.</dc:creator>
<dc:creator>Ganal-Vonarburg, S. C.</dc:creator>
<dc:creator>Grosjean, M.</dc:creator>
<dc:creator>Bigalke, M.</dc:creator>
<dc:creator>Spielvogel, S.</dc:creator>
<dc:creator>Macpherson, A.</dc:creator>
<dc:creator>Mestrot, A.</dc:creator>
<dc:creator>Hapfelmeier, S.</dc:creator>
<dc:creator>Erb, M.</dc:creator>
<dc:creator>Schlaeppi, K.</dc:creator>
<dc:creator>Ramette, A.</dc:creator>
<dc:date>2024-04-21</dc:date>
<dc:identifier>doi:10.1101/2024.04.20.590402</dc:identifier>
<dc:title><![CDATA[Component specific responses of the microbiomes to common chemical stressors in the human food chain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.24.590951v1?rss=1">
<title>
<![CDATA[
Braf-mutant Schwann cells divert to a repair phenotype to induce congenital demyelinating neuropathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.24.590951v1?rss=1</link>
<description><![CDATA[
RASopathies, rare congenital syndromes affecting multiple organ systems, often include peripheral neuropathy of unknown origin. While RASopathy-associated gene variants are proto-oncogenic, the impact of timing and mosaicism on pathogenicity remains poorly understood. Here, we investigate the links between Braf, a key mitogen-activated protein kinase (MAPK) effector, and peripheral neuropathy. By targeting Braf p.V600E, an oncogenic variant found in mosaic RASopathies, to embryonic Mpz-expressing cells in mice, we induced a congenital Charcot-Marie-Tooth-like degenerative neuropathy. This phenotype was characterized by hyperplastic nerves, hindlimb weakness, and unexpectedly reduced body size. Constitutively active Braf expanded a Jun+ Schwann cell repair state, impairing myelination and nerve homeostasis. To examine relevance to RASopathies, we differentiated patient-derived stem cells bearing the cardio-facio-cutaneous syndrome-associated BRAF p.Q257R variant into Schwann cells. Compared to wild-type controls, CFC-derived cells failed to acquire mature phenotypes, instead exhibiting progenitor or repair-type transcriptional profiles. Our findings implicate somatic mosaicism in the unresolved genetic heterogeneity of neuropathies and expand the candidate gene list for peripheral nerve disorders. Moreover, they reveal a MAPK-dependent mechanism linking neural crest-derived Schwann cell dysfunction to both body growth and nerve homeostasis, providing new insights into the mechanisms in RASopathy-associated neuropathy and potential therapeutic targets.
]]></description>
<dc:creator>Marechal, E.</dc:creator>
<dc:creator>Quintana, P.</dc:creator>
<dc:creator>Aldea, D. A.</dc:creator>
<dc:creator>Mondielli, G.</dc:creator>
<dc:creator>Bernard-Marissal, N.</dc:creator>
<dc:creator>Moreno, M.</dc:creator>
<dc:creator>Delague, V.</dc:creator>
<dc:creator>Weiss, L.</dc:creator>
<dc:creator>Barlier, A.</dc:creator>
<dc:creator>Etchevers, H. C.</dc:creator>
<dc:date>2024-04-25</dc:date>
<dc:identifier>doi:10.1101/2024.04.24.590951</dc:identifier>
<dc:title><![CDATA[Braf-mutant Schwann cells divert to a repair phenotype to induce congenital demyelinating neuropathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.25.591105v1?rss=1">
<title>
<![CDATA[
Estrogen Receptor alpha/14-3-3 molecular glues as alternative treatment strategy for endocrine resistant breast cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.25.591105v1?rss=1</link>
<description><![CDATA[
Endocrine resistance in breast cancer treatment is a major clinical hurdle, causing an urgent need for alternative treatment modalities. The suppressive protein-protein interaction (PPI) between Estrogen Receptor alpha (ER) and the adaptor protein 14-3-3 offers such a strategy. Here, we report the biological impact of small-molecule  molecular glues of this ER/14-3-3 PPI by using both fusicoccin-derived semi-synthetic natural products and fully synthetic covalent drug-like molecules. We show that the ER/14-3-3 PPI is stabilized by both the natural- and synthetic glues, resulting in a suppression of ER transcriptional activity and a blockade of breast cancer cell proliferation, both in cell lines and in organoids derived from endocrine therapy resistant breast cancer patients. Importantly, the molecular glues effectively blocked ER action even in case of constitutively active clinical ER mutations, providing the foundations for developing alternative classes of ER targeting compounds to improve treatment of patients with endocrine-therapy resistant breast cancer.



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]]></description>
<dc:creator>Visser, E.</dc:creator>
<dc:creator>Donaldson Collier, M.</dc:creator>
<dc:creator>Siefert, J.</dc:creator>
<dc:creator>Konstantinidou, M.</dc:creator>
<dc:creator>Paul, S.</dc:creator>
<dc:creator>Berkhout, J.</dc:creator>
<dc:creator>Virta, J.</dc:creator>
<dc:creator>Somsen, B.</dc:creator>
<dc:creator>Cossar, P.</dc:creator>
<dc:creator>Miley, G.</dc:creator>
<dc:creator>Luzietti, L.</dc:creator>
<dc:creator>Young, L.</dc:creator>
<dc:creator>Vareslija, D.</dc:creator>
<dc:creator>Buluwela, L.</dc:creator>
<dc:creator>Ali, S.</dc:creator>
<dc:creator>Meijer, O.</dc:creator>
<dc:creator>Arkin, M. R.</dc:creator>
<dc:creator>Ottmann, C. O. R.</dc:creator>
<dc:creator>Zwart, W.</dc:creator>
<dc:creator>Brunsveld, L.</dc:creator>
<dc:date>2024-04-27</dc:date>
<dc:identifier>doi:10.1101/2024.04.25.591105</dc:identifier>
<dc:title><![CDATA[Estrogen Receptor alpha/14-3-3 molecular glues as alternative treatment strategy for endocrine resistant breast cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.25.591130v1?rss=1">
<title>
<![CDATA[
LSD flattens the hierarchy of directed information flow in fast whole-brain dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.25.591130v1?rss=1</link>
<description><![CDATA[
Psychedelics are serotonergic drugs that profoundly alter consciousness, yet their neural mechanisms are not fully understood. A popular theory, RElaxed Beliefs Under pSychedelics (REBUS), posits that psychedelics flatten the hierarchy of information flow in the brain. Here, we investigate hierarchy based on the imbalance between sending and receiving brain signals, as determined by directed functional connectivity. We measure directed functional hierarchy in a magnetoencephalography (MEG) dataset of 16 healthy human participants who were administered a psychedelic dose (75 micrograms, intravenous) of lysergic acid diethylamide (LSD) under four different conditions. LSD diminishes the asymmetry of directed connectivity when averaged across time. Additionally, we demonstrate that machine learning classifiers distinguish between LSD and placebo more accurately when trained on one of our hierarchy metrics than when trained on traditional measures of functional connectivity. Taken together, these results indicate that LSD weakens the hierarchy of directed connectivity in the brain by increasing the balance between senders and receivers of neural signals.
]]></description>
<dc:creator>Shinozuka, K.</dc:creator>
<dc:creator>Tewarie, P.</dc:creator>
<dc:creator>Luppi, A.</dc:creator>
<dc:creator>Lynn, C.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Muthukumaraswamy, S. D.</dc:creator>
<dc:creator>Nutt, D. J.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Deco, G.</dc:creator>
<dc:creator>Kringelbach, M. L.</dc:creator>
<dc:date>2024-04-28</dc:date>
<dc:identifier>doi:10.1101/2024.04.25.591130</dc:identifier>
<dc:title><![CDATA[LSD flattens the hierarchy of directed information flow in fast whole-brain dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.25.591183v1?rss=1">
<title>
<![CDATA[
Microstructural Mapping of Neural Pathways in Alzheimer's Disease using Macrostructure-Informed Normative Tractometry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.25.591183v1?rss=1</link>
<description><![CDATA[
IntroductionDiffusion MRI is sensitive to the microstructural properties of brain tissues, and shows great promise in detecting the effects of degenerative diseases. However, many approaches analyze single measures averaged over regions of interest, without considering the underlying fiber geometry.

MethodsHere, we propose a novel Macrostructure-Informed Normative Tractometry (MINT) framework, to investigate how white matter microstructure and macrostructure are jointly altered in mild cognitive impairment (MCI) and dementia. We compare MINT-derived metrics with univariate metrics from diffusion tensor imaging (DTI), to examine how fiber geometry may impact interpretation of microstructure.

ResultsIn two multi-site cohorts from North America and India, we find consistent patterns of microstructural and macrostructural anomalies implicated in MCI and dementia; we also rank diffusion metrics sensitivity to dementia.

DiscussionWe show that MINT, by jointly modeling tract shape and microstructure, has potential to disentangle and better interpret the effects of degenerative disease on the brains neural pathways.
]]></description>
<dc:creator>Feng, Y.</dc:creator>
<dc:creator>Chandio, B. Q.</dc:creator>
<dc:creator>Villalon-Reina, J. E.</dc:creator>
<dc:creator>Thomopoulos, S. I.</dc:creator>
<dc:creator>Nir, T. M.</dc:creator>
<dc:creator>Benavidez, S.</dc:creator>
<dc:creator>Laltoo, E.</dc:creator>
<dc:creator>Chattopadhyay, T.</dc:creator>
<dc:creator>Joshi, H.</dc:creator>
<dc:creator>Venkatasubramanian, G.</dc:creator>
<dc:creator>John, J. P.</dc:creator>
<dc:creator>Jahanshad, N.</dc:creator>
<dc:creator>Reid, R. I.</dc:creator>
<dc:creator>Jack, C. R.</dc:creator>
<dc:creator>Weiner, M. M.</dc:creator>
<dc:creator>Thompson, P. M.</dc:creator>
<dc:creator>the Alzheimer's Disease Neuroimaging Initiative,</dc:creator>
<dc:date>2024-04-28</dc:date>
<dc:identifier>doi:10.1101/2024.04.25.591183</dc:identifier>
<dc:title><![CDATA[Microstructural Mapping of Neural Pathways in Alzheimer's Disease using Macrostructure-Informed Normative Tractometry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.28.591397v1?rss=1">
<title>
<![CDATA[
Unifying community-wide whole-brain imaging datasets enables robust automated neuron identification and reveals determinants of neuron positioning in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.28.591397v1?rss=1</link>
<description><![CDATA[
We develop a data harmonization approach for C. elegans volumetric microscopy data, still or video, consisting of a standardized format, data pre-processing techniques, and a set of human-in-the-loop machine learning based analysis software tools. We unify a diverse collection of 118 whole-brain neural activity imaging datasets from 5 labs, storing these and accompanying tools in an online repository called WormID (wormid.org). We use this repository to train three existing automated cell identification algorithms to, for the first time, enable accuracy in neural identification that generalizes across labs, approaching human performance in some cases. We mine this repository to identify factors that influence the developmental positioning of neurons. To facilitate communal use of this repository, we created open-source software, code, web-based tools, and tutorials to explore and curate datasets for contribution to the scientific community. This repository provides a growing resource for experimentalists, theorists, and toolmakers to (a) study neuroanatomical organization and neural activity across diverse experimental paradigms, (b) develop and benchmark algorithms for automated neuron detection, segmentation, cell identification, tracking, and activity extraction, and (c) inform models of neurobiological development and function.
]]></description>
<dc:creator>Sprague, D. Y.</dc:creator>
<dc:creator>Rusch, K.</dc:creator>
<dc:creator>Dunn, R. L.</dc:creator>
<dc:creator>Borchardt, J.</dc:creator>
<dc:creator>Bubnis, G.</dc:creator>
<dc:creator>Chiu, G.</dc:creator>
<dc:creator>Wen, C.</dc:creator>
<dc:creator>Suzuki, R.</dc:creator>
<dc:creator>Chaudhary, S.</dc:creator>
<dc:creator>Dichter, B.</dc:creator>
<dc:creator>Ly, R.</dc:creator>
<dc:creator>Onami, S.</dc:creator>
<dc:creator>Lu, H.</dc:creator>
<dc:creator>Kimura, K.</dc:creator>
<dc:creator>Yemini, E. I.</dc:creator>
<dc:creator>Kato, S.</dc:creator>
<dc:date>2024-04-30</dc:date>
<dc:identifier>doi:10.1101/2024.04.28.591397</dc:identifier>
<dc:title><![CDATA[Unifying community-wide whole-brain imaging datasets enables robust automated neuron identification and reveals determinants of neuron positioning in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.29.591704v1?rss=1">
<title>
<![CDATA[
P66 is a bacterial mimic of CD47 that binds the anti-phagocytic receptor SIRPα and facilitates macrophage evasion by Borrelia burgdorferi 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.29.591704v1?rss=1</link>
<description><![CDATA[
Summary ParagraphInnate immunity, the first line of defense against pathogens, relies on efficient elimination of invading agents by phagocytes. In the co-evolution of host and pathogen, pathogens developed mechanisms to dampen and evade phagocytic clearance. Here, we report that bacterial pathogens can evade clearance by macrophages through mimicry at the mammalian anti-phagocytic "dont eat me" signaling axis between CD47 (ligand) and SIRP (receptor). We identified a protein, P66, on the surface of Borrelia burgdorferi that, like CD47, is necessary and sufficient to bind the macrophage receptor SIRP. Expression of the gene encoding the protein is required for bacteria to bind SIRP or a high-affinity CD47 reagent. Genetic deletion of p66 increases phagocytosis by macrophages. Blockade of P66 during infection promotes clearance of the bacteria. This study demonstrates that mimicry of the mammalian anti-phagocytic protein CD47 by B. burgdorferi inhibits macrophage-mediated bacterial clearance. Such a mechanism has broad implications for understanding of host-pathogen interactions and expands the function of the established innate immune checkpoint receptor SIRP. Moreover, this report reveals P66 as a novel therapeutic target in the treatment of Lyme Disease.
]]></description>
<dc:creator>Tal, M. C.</dc:creator>
<dc:creator>Hansen, P. S.</dc:creator>
<dc:creator>Ogasawara, H. A.</dc:creator>
<dc:creator>Feng, Q.</dc:creator>
<dc:creator>Volk, R. F.</dc:creator>
<dc:creator>Lee, B.</dc:creator>
<dc:creator>Casebeer, S. E.</dc:creator>
<dc:creator>Blacker, G. S.</dc:creator>
<dc:creator>Shoham, M.</dc:creator>
<dc:creator>Galloway, S. D.</dc:creator>
<dc:creator>Sapiro, A. L.</dc:creator>
<dc:creator>Dulgeroff, L.-B. T.</dc:creator>
<dc:creator>Raveh, T.</dc:creator>
<dc:creator>Pothineni, V. R.</dc:creator>
<dc:creator>Potula, H.-H. S.</dc:creator>
<dc:creator>Rajadas, J.</dc:creator>
<dc:creator>Bastounis, E. E.</dc:creator>
<dc:creator>Chou, S.</dc:creator>
<dc:creator>Robinson, W. H.</dc:creator>
<dc:creator>Coburn, J.</dc:creator>
<dc:creator>Weissman, I. L.</dc:creator>
<dc:creator>Zaro, B. W.</dc:creator>
<dc:date>2024-04-30</dc:date>
<dc:identifier>doi:10.1101/2024.04.29.591704</dc:identifier>
<dc:title><![CDATA[P66 is a bacterial mimic of CD47 that binds the anti-phagocytic receptor SIRPα and facilitates macrophage evasion by Borrelia burgdorferi]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.01.591950v1?rss=1">
<title>
<![CDATA[
An shRNA screen in primary human beta cells identifies the serotonin 1F receptor as a negative regulator of survival during transplant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.01.591950v1?rss=1</link>
<description><![CDATA[
Islet transplantation can cure type 1 diabetes, but peri-transplant beta cell death limits this procedure to those with low insulin requirements. Improving human beta cell survival or proliferation may make islet transplantation a possibility for more type 1 patients. To identify novel regulators of beta cell survival and proliferation, we conducted a pooled small hairpin RNA (shRNA) screen in primary human beta cells transplanted into immunocompromised mice. shRNAs targeting several cyclin dependent kinase inhibitors were enriched after transplant. Here, we focused on the Gi/o-coupled GPCR, serotonin 1F receptor (HTR1F, 5-HT1F) which our screen identified as a negative regulator of beta cell numbers after transplant. In vitro, 5-HT1F knockdown induced human beta cell proliferation but only when combined with harmine and exendin-4. In vivo, knockdown of 5-HT1F reduced beta cell death during transplant. To demonstrate the feasibility of targeting 5-HT1F in islet transplant, we identified and validated a small molecule 5-HT1F antagonist. This antagonist increased glucose stimulated insulin secretion from primary human islets and cAMP accumulation in primary human beta cells. Finally, the 5-HT1F antagonist improved glycemia in marginal mass, human islet transplants into immunocompromised mice. We identify 5-HT1F as a novel druggable target to improve human beta cell survival in the setting of islet transplantation.

One Sentence SummarySerotonin 1F receptor (5-HT1F) negatively regulates insulin secretion and beta cell survival during transplant.
]]></description>
<dc:creator>Lee, R. A.</dc:creator>
<dc:creator>Chopra, D. G.</dc:creator>
<dc:creator>Nguyen, V.</dc:creator>
<dc:creator>Huang, X.-P.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Shariati, K.</dc:creator>
<dc:creator>Yiv, N.</dc:creator>
<dc:creator>Schugar, R.</dc:creator>
<dc:creator>Annes, J.</dc:creator>
<dc:creator>Roth, B. L.</dc:creator>
<dc:creator>Ku, G.</dc:creator>
<dc:date>2024-05-03</dc:date>
<dc:identifier>doi:10.1101/2024.05.01.591950</dc:identifier>
<dc:title><![CDATA[An shRNA screen in primary human beta cells identifies the serotonin 1F receptor as a negative regulator of survival during transplant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.05.592598v1?rss=1">
<title>
<![CDATA[
Genetic variants associated with cell-type-specific intra-individual gene expression variability reveal new mechanisms of genome regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.05.592598v1?rss=1</link>
<description><![CDATA[
Gene expression levels can vary substantially across cells, even in a seemingly homogeneous cell population. Identifying the relationships between genetic variation and gene expression is critical for understanding the mechanisms of genome regulation. However, the genetic control of gene expression variability among the cells within individuals has yet to be extensively examined. This is primarily due to the statistical challenges, such as the need for sufficiently powered cohorts and adjusting mean-variance dependence. Here, we introduce MEOTIVE (Mapping genetic Effects On inTra-Individual Variability of gene Expression), a novel statistical framework to identify genetic effects on the gene expression variability (sc-veQTL) accounting for the mean-variance dependence. Using single-cell RNA-seq data of 1.2 million peripheral blood mononuclear cells from 980 human donors, we identified 14 - 3,488 genes with significant sc-veQTLs (study-wide q-value < 0.05) across different blood cell types, 2,103 of which were shared across more than one cell type. We further detected 55 SNP-gene pairs (in 34 unique genes) by directly linking genetic variations with gene expression dispersion (sc-deQTL) regardless of mean-variance dependence, and these genes were enriched in biological processes relevant to immune response and viral infection. An example is rs1131017 (p<9.08x10-52), a sc-veQTL in the 5UTR of RPS26, which shows a ubiquitous dispersion effect across cell types, with higher dispersion levels associated with lower auto-immune disease risk, including rheumatoid arthritis and type 1 diabetes. Another example is LYZ, which is associated with antibacterial activity against bacterial species and was only detected with a monocyte-specific deQTL (rs1384) located at the 3 UTR region (p=1.48x10-11) and replicated in an independent cohort. Our results demonstrate an efficient and robust statistical method to identify genetic effects on gene expression variability and how these associations and their involved pathways confer auto-immune disease risk. This analytical framework provides a new approach to unravelling the genetic regulation of gene expression at the single-cell resolution, advancing our understanding of complex biological processes.
]]></description>
<dc:creator>Powell, J. E.</dc:creator>
<dc:creator>Xue, A.</dc:creator>
<dc:creator>Yazar, S.</dc:creator>
<dc:creator>Alquicira, J.</dc:creator>
<dc:creator>Cuomo, A.</dc:creator>
<dc:creator>Senabouth, A.</dc:creator>
<dc:creator>Gordon, G.</dc:creator>
<dc:creator>Kathail, P.</dc:creator>
<dc:creator>Ye, J.</dc:creator>
<dc:creator>Hewitt, A.</dc:creator>
<dc:date>2024-05-07</dc:date>
<dc:identifier>doi:10.1101/2024.05.05.592598</dc:identifier>
<dc:title><![CDATA[Genetic variants associated with cell-type-specific intra-individual gene expression variability reveal new mechanisms of genome regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.05.592608v1?rss=1">
<title>
<![CDATA[
Data-driven fine-grained region discovery in the mouse brain with transformers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.05.592608v1?rss=1</link>
<description><![CDATA[
Spatial transcriptomics offers unique opportunities to define the spatial organization of tissues and organs, such as the mouse brain. We address a key bottleneck in the analysis of organ-scale spatial transcriptomic data by establishing a workflow for self-supervised spatial domain detection that is scalable to multimillion cell datasets. This workflow uses a self-supervised framework for learning latent representations of tissue spatial domains or niches. We use a novel encoder-decoder architecture, which we named CellTransformer, to hierarchically learn higher-order tissue features from lower-level cellular and molecular statistical patterns. Coupling our representation learning workflow with minibatched GPU-accelerated clustering algorithms allows us to scale to multi-million cell MERFISH datasets where other methods cannot. CellTransformer is effective at integrating cells across tissue sections, identifying domains highly similar to ones in existing ontologies such as Allen Mouse Brain Common Coordinate Framework (CCF) while allowing discovery of hundreds of uncataloged areas with minimal loss of domain spatial coherence. CellTransformer domains recapitulate previous neuroanatomical studies of areas in the subiculum and superior colliculus, and characterize putatively uncataloged subregions in subcortical areas which currently lack subregion annotation. CellTransformer is also capable of domain discovery in whole-brain Slide-seqV2 datasets. Our workflows enable complex multi-animal analyses, achieving nearly perfect consistency of up to 100 spatial domains in a dataset of four individual mice with nine million cells across more than 200 tissue sections. CellTransformer advances the state of the art for spatial transcriptomics, by providing a performant solution for detection of fine-grained tissue domains from spatial transcriptomics data.
]]></description>
<dc:creator>Lee, A. J.</dc:creator>
<dc:creator>Yao, S.</dc:creator>
<dc:creator>Lusk, N.</dc:creator>
<dc:creator>Zeng, H.</dc:creator>
<dc:creator>Tasic, B.</dc:creator>
<dc:creator>Abbasi-Asl, R.</dc:creator>
<dc:date>2024-05-07</dc:date>
<dc:identifier>doi:10.1101/2024.05.05.592608</dc:identifier>
<dc:title><![CDATA[Data-driven fine-grained region discovery in the mouse brain with transformers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.06.592854v1?rss=1">
<title>
<![CDATA[
NEK1 modulates neurite outgrowth in motor neurons through coordinating retromer formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.06.592854v1?rss=1</link>
<description><![CDATA[
Loss of function of a cell cycle-associated gene NEK1 causes amyotrophic lateral sclerosis (ALS), but how this leads to motor neuron degeneration is unknown. We studied the function of NEK1 in human stem cell-derived motor neurons and found that loss of NEK1 causes decreased neurite length accompanied by transcriptional alterations. We also found that NEK1 interacts with and modulates the formation of the retromer, and that impaired retromer function contributes to neurite outgrowth deficits. We identified SMC3, which interacts with NEK1 during the cell cycle, as a kinase substrate of NEK1 in motor neurons. Notably, loss of SMC3 not only recapitulates the decreased neurite outgrowth, but also affects retromer formation. We suggest that NEK1 interacts with multiple proteins in postmitotic neurons to coordinate retromer formation, and that loss of this leads to impaired neurite outgrowth.
]]></description>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Oses, J. A.</dc:creator>
<dc:creator>Kawaguchi, R.</dc:creator>
<dc:creator>Perrault, L.</dc:creator>
<dc:creator>Frost, D.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Guo, F.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Zhao, P.</dc:creator>
<dc:creator>Ho, T. S.-y.</dc:creator>
<dc:creator>Pandey, R.</dc:creator>
<dc:creator>Geschwind, D. H.</dc:creator>
<dc:creator>Burlingame, A.</dc:creator>
<dc:creator>Woolf, C.</dc:creator>
<dc:date>2024-05-08</dc:date>
<dc:identifier>doi:10.1101/2024.05.06.592854</dc:identifier>
<dc:title><![CDATA[NEK1 modulates neurite outgrowth in motor neurons through coordinating retromer formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.08.593085v1?rss=1">
<title>
<![CDATA[
Closed-loop modulation of remote hippocampal representations with neurofeedback 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.08.593085v1?rss=1</link>
<description><![CDATA[
Humans can remember specific remote events without acting on them and influence which memories are retrieved based on internal goals. However, animal models typically present sensory cues to trigger memory retrieval and then assess retrieval based on action. Thus, it is difficult to determine whether measured neural activity patterns relate to the cue(s), the memory, or the behavior. We therefore asked whether retrieval-related neural activity could be generated in animals without cues or a behavioral report. We focused on hippocampal "place cells" which primarily represent the animals current location (local representations) but can also represent locations away from the animal (remote representations). We developed a neurofeedback system to reward expression of remote representations and found that rats could learn to generate specific spatial representations that often jumped directly to the experimenter-defined target location. Thus, animals can deliberately engage remote representations, enabling direct study of retrieval-related activity in the brain.
]]></description>
<dc:creator>Coulter, M. E.</dc:creator>
<dc:creator>Gillespie, A. K.</dc:creator>
<dc:creator>Chu, J. P.</dc:creator>
<dc:creator>Denovellis, E. L.</dc:creator>
<dc:creator>Nguyen, T. T. K.</dc:creator>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Wadhwani, K.</dc:creator>
<dc:creator>Sharma, B.</dc:creator>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Eden, U.</dc:creator>
<dc:creator>Kemere, C.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:date>2024-05-08</dc:date>
<dc:identifier>doi:10.1101/2024.05.08.593085</dc:identifier>
<dc:title><![CDATA[Closed-loop modulation of remote hippocampal representations with neurofeedback]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.09.593336v1?rss=1">
<title>
<![CDATA[
A mismatch between striatal cholinergic pauses and dopaminergic reward prediction errors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.09.593336v1?rss=1</link>
<description><![CDATA[
Movement, motivation and reward-related learning depend strongly on striatal dopamine and acetylcholine. These neuromodulators each regulate the other, and disturbances to their coordinated signals contribute to human disorders ranging from Parkinsons Disease to depression and addiction. Pauses in the firing of cholinergic interneurons (CINs) are thought to coincide with pulses in dopamine release that encode reward prediction errors (RPEs), together shaping synaptic plasticity and thereby learning. However, such models are based upon recordings from unidentified neurons, and do not incorporate the distinct characteristics of striatal subregions. Here we compare the firing of identified, individual CINs to dopamine release as unrestrained rats performed a probabilistic decision-making task. The relationships between CIN spiking, dopamine release, and behavior varied strongly by subregion. In dorsal-lateral striatum a Go! cue evoked burst-pause CIN spiking, quickly followed by a very brief (~150ms) dopamine pulse that was unrelated to RPE. In dorsal-medial striatum the same cue evoked only a CIN pause; this pause was curtailed by a movement-selective rebound in firing. Finally in ventral striatum a reward cue evoked slower, RPE-coding increases in both dopamine and CIN firing, without any distinct pause. Our results demonstrate a spatial and temporal dissociation between CIN pauses and dopamine RPE signals, and will inform new models of striatal microcircuits and their contributions to behavior.
]]></description>
<dc:creator>Duhne, M.</dc:creator>
<dc:creator>Mohebi, A.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Pelattini, L.</dc:creator>
<dc:creator>Berke, J. D.</dc:creator>
<dc:date>2024-05-09</dc:date>
<dc:identifier>doi:10.1101/2024.05.09.593336</dc:identifier>
<dc:title><![CDATA[A mismatch between striatal cholinergic pauses and dopaminergic reward prediction errors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.07.592993v1?rss=1">
<title>
<![CDATA[
Indels allow antiviral proteins to evolve functional novelty inaccessible by missense mutations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.07.592993v1?rss=1</link>
<description><![CDATA[
Antiviral proteins often evolve rapidly at virus-binding interfaces to defend against new viruses. We investigated whether antiviral adaptation via missense mutations might face limits, which insertion or deletion mutations (indels) could overcome. We report one such case of a nearly insurmountable evolutionary challenge: the human anti-retroviral protein TRIM5 requires more than five missense mutations in its specificity-determining v1 loop to restrict a divergent simian immunodeficiency virus (SIV). However, duplicating just one amino acid in v1 enables human TRIM5 to potently restrict SIV in a single evolutionary step. Moreover, natural primate TRIM5 v1 loops have evolved indels that confer novel antiviral specificities. Thus, indels enable antiviral proteins to overcome viral challenges inaccessible by missense mutations, revealing the potential of these often-overlooked mutations in driving protein innovation.
]]></description>
<dc:creator>Tenthorey, J.</dc:creator>
<dc:creator>del Banco, S.</dc:creator>
<dc:creator>Ramzan, I.</dc:creator>
<dc:creator>Klingenberg, H.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2024-05-10</dc:date>
<dc:identifier>doi:10.1101/2024.05.07.592993</dc:identifier>
<dc:title><![CDATA[Indels allow antiviral proteins to evolve functional novelty inaccessible by missense mutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.12.593711v1?rss=1">
<title>
<![CDATA[
Endogenous OptoRhoGEFs reveal biophysical principles of epithelial tissue furrowing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.12.593711v1?rss=1</link>
<description><![CDATA[
During development, epithelia function as malleable substrates that undergo extensive remodeling to shape developing embryos. Optogenetic control of Rho signaling provides an avenue to investigate the mechanisms of epithelial morphogenesis, but transgenic optogenetic tools can be limited by variability in tool expression levels and deleterious effects of transgenic overexpression on development. Here, we use CRISPR/Cas9 to tag Drosophila RhoGEF2 and Cysts/Dp114RhoGEF with components of the iLID/SspB optogenetic heterodimer, permitting light-dependent control over endogenous protein activities. Using quantitative optogenetic perturbations, we uncover a dose-dependence of tissue furrow depth and bending behavior on RhoGEF recruitment, revealing mechanisms by which developing embryos can shape tissues into particular morphologies. We show that at the onset of gastrulation, furrows formed by cell lateral contraction are oriented and size-constrained by a stiff basal actomyosin layer. Our findings demonstrate the use of quantitative, 3D-patterned perturbations of cell contractility to precisely shape tissue structures and interrogate developmental mechanics.
]]></description>
<dc:creator>Countryman, A. D.</dc:creator>
<dc:creator>Doherty, C. A.</dc:creator>
<dc:creator>Herrera-Perez, R. M.</dc:creator>
<dc:creator>Kasza, K. E.</dc:creator>
<dc:date>2024-05-12</dc:date>
<dc:identifier>doi:10.1101/2024.05.12.593711</dc:identifier>
<dc:title><![CDATA[Endogenous OptoRhoGEFs reveal biophysical principles of epithelial tissue furrowing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.09.592452v1?rss=1">
<title>
<![CDATA[
In Utero Hematopoietic Stem Cell Transplant for Fanconi Anemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.09.592452v1?rss=1</link>
<description><![CDATA[
Fanconi Anemia (FA) is an inherited DNA-repair deficiency caused by mutations in diverse Fanc genes that leads to bone marrow failure and malignancies. FA disease begins at early embryonic stages, and while FA prenatal testing has long been available, no fetal therapies for FA currently exist. Postnatally, FA hematologic disease can be cured through allogeneic hematopoietic stem cell transplantation (HSCT); however, this requires chemotherapy and/or irradiation-based conditioning which amongst various side-effects also increases likelihood of malignancies later in life in these fragile patients. Given fetal immune tolerance and the competitive advantage of healthy hematopoietic stem and progenitor cells (HSPCs) over failing FA HSPCs, in utero HSCT without conditioning may be an alternative approach to stabilization of the hematopoietic system without conventional toxicities. We performed in utero HSCT using HSPCs from wildtype (WT) donors into two FA mouse models (Fancd2-/-, Fanca-/-) and observed robust multi-lineage hematopoietic donor engraftment in homozygous FA mice compared to both heterozygous FA and WT littermates. Upon serial assessments, we also observed increasing donor chimerism up to 94.1%, showcasing the competitive advantage of WT donor HSPCs over FA HSPCs. Given that 1% donor chimerism is predicted to stabilize FA BM, in utero HSCT may be a safe and curative prenatal treatment for all subtypes of FA.
]]></description>
<dc:creator>Swartzrock, L.</dc:creator>
<dc:creator>Dib, C.</dc:creator>
<dc:creator>Denis, M.</dc:creator>
<dc:creator>Willner, H.</dc:creator>
<dc:creator>Ho, K.</dc:creator>
<dc:creator>Haslett, E.</dc:creator>
<dc:creator>Krampf, M. R.</dc:creator>
<dc:creator>Girsen, A.</dc:creator>
<dc:creator>Blumenfeld, Y. J.</dc:creator>
<dc:creator>El-Sayed, Y. Y.</dc:creator>
<dc:creator>Roncarolo, M. G.</dc:creator>
<dc:creator>MacKenzie, T. C.</dc:creator>
<dc:creator>Czechowicz, A. D.</dc:creator>
<dc:date>2024-05-14</dc:date>
<dc:identifier>doi:10.1101/2024.05.09.592452</dc:identifier>
<dc:title><![CDATA[In Utero Hematopoietic Stem Cell Transplant for Fanconi Anemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.10.593451v1?rss=1">
<title>
<![CDATA[
Pro-inflammatory macrophage activation does not require inhibition of mitochondrial respiration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.10.593451v1?rss=1</link>
<description><![CDATA[
Pro-inflammatory macrophage activation is a hallmark example of how mitochondria serve as signaling organelles. Upon classical macrophage activation, oxidative phosphorylation sharply decreases and mitochondria are repurposed to accumulate signals that amplify effector function. However, evidence is conflicting as to whether this collapse in respiration is essential or largely dispensable. Here we systematically examine this question and show that reduced oxidative phosphorylation is not required for pro-inflammatory macrophage activation. Only stimuli that engage both MyD88- and TRIF-linked pathways decrease mitochondrial respiration, and different pro-inflammatory stimuli have varying effects on other bioenergetic parameters. Additionally, pharmacologic and genetic models of electron transport chain inhibition show no direct link between respiration and pro-inflammatory activation. Studies in mouse and human macrophages also reveal accumulation of the signaling metabolites succinate and itaconate can occur independently of characteristic breaks in the TCA cycle. Finally, in vivo activation of peritoneal macrophages further demonstrates that a pro-inflammatory response can be elicited without reductions to oxidative phosphorylation. Taken together, the results suggest the conventional model of mitochondrial reprogramming upon macrophage activation is incomplete.
]]></description>
<dc:creator>Ball, A. B.</dc:creator>
<dc:creator>Jones, A. E.</dc:creator>
<dc:creator>Nguyen, K. B.</dc:creator>
<dc:creator>Rios, A.</dc:creator>
<dc:creator>Marx, N.</dc:creator>
<dc:creator>Hsieh, W. Y.</dc:creator>
<dc:creator>Yang, K.</dc:creator>
<dc:creator>Desousa, B. R.</dc:creator>
<dc:creator>Kim, K. K. O.</dc:creator>
<dc:creator>Veliova, M.</dc:creator>
<dc:creator>del Mundo, Z. M.</dc:creator>
<dc:creator>Shirihai, O. S.</dc:creator>
<dc:creator>Beninca, C.</dc:creator>
<dc:creator>Stiles, L.</dc:creator>
<dc:creator>Bensinger, S. J.</dc:creator>
<dc:creator>Divakaruni, A. S.</dc:creator>
<dc:date>2024-05-14</dc:date>
<dc:identifier>doi:10.1101/2024.05.10.593451</dc:identifier>
<dc:title><![CDATA[Pro-inflammatory macrophage activation does not require inhibition of mitochondrial respiration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.11.593442v1?rss=1">
<title>
<![CDATA[
Sexually dimorphic regulatory T cell-derived enkephalin imparts pregnancy-induced analgesia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.11.593442v1?rss=1</link>
<description><![CDATA[
T cells have emerged as sex-dependent orchestrators of pain chronification but the sexually dimorphic mechanisms by which T cells control pain sensitivity is not resolved. Here, we demonstrate an influence of regulatory T cells (Tregs) on pain processing that is distinct from their canonical functions of immune regulation and tissue repair. Specifically, meningeal Tregs (mTregs) express the endogenous opioid, enkephalin, and mTreg-derived enkephalin exerts an antinociceptive action through a presynaptic opioid receptor signaling mechanism that is dispensable for immunosuppression. We demonstrate that mTregs are both necessary and sufficient to suppress mechanical pain sensitivity in female, but not male, mice, with this modulation reliant on sex hormones. These results uncover a fundamental sex-specific, and immunologically- derived endogenous opioid circuit for nociceptive regulation with critical implications for pain biology.

Highlights1. Gating of allodynia by meningeal Tregs is sex hormone-dependent

3. Treg-derived enkephalin modulates mechanical pain sensitivity, not inflammation

4. Delta opioid receptor on MrgprD+ sensory neuron mediates pain processing by mTregs
]]></description>
<dc:creator>Midavaine, E.</dc:creator>
<dc:creator>Moraes, B. C.</dc:creator>
<dc:creator>Benitez, J.</dc:creator>
<dc:creator>Rodriguez, S. R.</dc:creator>
<dc:creator>Braz, J. M.</dc:creator>
<dc:creator>Kochhar, N. P.</dc:creator>
<dc:creator>Eckalbar, W. L.</dc:creator>
<dc:creator>Domingos, A. I.</dc:creator>
<dc:creator>Pintar, J. E.</dc:creator>
<dc:creator>Basbaum, A. I.</dc:creator>
<dc:creator>Kashem, S. W.</dc:creator>
<dc:date>2024-05-14</dc:date>
<dc:identifier>doi:10.1101/2024.05.11.593442</dc:identifier>
<dc:title><![CDATA[Sexually dimorphic regulatory T cell-derived enkephalin imparts pregnancy-induced analgesia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.12.592631v1?rss=1">
<title>
<![CDATA[
Structural basis of autoantibody binding in Neuromyelitis Optica 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.12.592631v1?rss=1</link>
<description><![CDATA[
Neuromyelitis Optica (NMO) is an autoimmune disease of the central nervous system where pathogenic autoantibodies target the human astrocyte water channel aquaporin-4 causing neurological impairment. Autoantibody binding leads to complement dependent and complement independent cytotoxicity, ultimately resulting in astrocyte death, demyelination, and neuronal loss. Aquaporin-4 assembles in astrocyte plasma membranes as symmetric tetramers or as arrays of tetramers. We report molecular structures of aquaporin-4 alone and bound to Fab fragments from patient-derived NMO autoantibodies using cryogenic electron microscopy. Each antibody binds to epitopes comprised of three extracellular loops of aquaporin-4 with contributions from multiple molecules in the assembly. The structures distinguish between antibodies that bind to the tetrameric form of aquaporin-4, and those targeting higher order orthogonal arrays of tetramers that provide more diverse bridging epitopes.

One-Sentence SummaryPathogenic autoantibodies in Neuromyelitis Optica distinguish multiply presented epitopes on arrays of human aquaporin-4.
]]></description>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Khandelwal, N. K.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Hwang, P.</dc:creator>
<dc:creator>Pourmal, S.</dc:creator>
<dc:creator>Bennett, J. L.</dc:creator>
<dc:creator>Stroud, R. M.</dc:creator>
<dc:date>2024-05-14</dc:date>
<dc:identifier>doi:10.1101/2024.05.12.592631</dc:identifier>
<dc:title><![CDATA[Structural basis of autoantibody binding in Neuromyelitis Optica]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.14.594195v1?rss=1">
<title>
<![CDATA[
Stress hyperglycemia exacerbates inflammatory brain injury after stroke 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.14.594195v1?rss=1</link>
<description><![CDATA[
Post-stroke hyperglycemia occurs in 30% - 60% of ischemic stroke patients as part of the systemic stress response, but neither clinical evidence nor pre-clinical studies indicate whether post-stroke hyperglycemia affects stroke outcome. Here we investigated this issue using a mouse model of permanent ischemia. Mice were maintained either normoglycemic or hyperglycemic during the interval of 17 - 48 hours after ischemia onset. Post-stroke hyperglycemia was found to increase infarct volume, blood-brain barrier disruption, and hemorrhage formation, and to impair motor recovery. Post-stroke hyperglycemia also increased superoxide formation by peri-infarct microglia/macrophages. In contrast, post-stroke hyperglycemia did not increase superoxide formation or exacerbate motor impairment in p47phox-/- mice, which cannot form an active superoxide-producing NADPH oxidase-2 complex. These results suggest that hyperglycemia occurring hours-to-days after ischemia can increase oxidative stress in peri-infarct tissues by fueling NADPH oxidase activity in reactive microglia/macrophages, and by this mechanism contribute to worsened functional outcome.
]]></description>
<dc:creator>Won, S. J.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Butler, N. J.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Mocanu, E.</dc:creator>
<dc:creator>Nzoutchoum, O. T.</dc:creator>
<dc:creator>Lakkaraju, R.</dc:creator>
<dc:creator>Davis, J.</dc:creator>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Swanson, R. A.</dc:creator>
<dc:date>2024-05-14</dc:date>
<dc:identifier>doi:10.1101/2024.05.14.594195</dc:identifier>
<dc:title><![CDATA[Stress hyperglycemia exacerbates inflammatory brain injury after stroke]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.16.594509v1?rss=1">
<title>
<![CDATA[
Likelihood-based interactive local docking into cryo-EM maps in ChimeraX 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.16.594509v1?rss=1</link>
<description><![CDATA[
The interpretation of cryo-EM maps often includes the docking of known or predicted structures of the components, which is particularly useful when the map resolution is worse than 4 [A]. Although it can be effective to search the entire map to find the best placement of a component, the process can be slow when the maps are large. However, frequently there is a well-founded hypothesis about where particular components are located. In such cases, a local search using a map subvolume will be much faster because the search volume is smaller, and more sensitive because optimizing the search volume for the rotation search step enhances signal-to-noise. A Fourier-space likelihood-based local search approach, based on the previously-published em_placement software, has been implemented in the new emplace_local program. Tests confirm that the local search approach enhances speed and sensitivity of the computations. An interactive graphical interface in the ChimeraX molecular graphics program provides a convenient way to set up and evaluate docking calculations, particularly in defining the part of the map into which the components should be placed.

SynopsisLikelihood-based cryo-EM docking using our emplace_local software is faster and more sensitive than our related software, em_placement, when the approximate location of a component is known, and is available conveniently through a plugin to the ChimeraX visualization software.
]]></description>
<dc:creator>Read, R. J.</dc:creator>
<dc:creator>Pettersen, E. F.</dc:creator>
<dc:creator>McCoy, A. J.</dc:creator>
<dc:creator>Croll, T. I.</dc:creator>
<dc:creator>Terwilliger, T. C.</dc:creator>
<dc:creator>Poon, B. K.</dc:creator>
<dc:creator>Meng, E. C.</dc:creator>
<dc:creator>Liebschner, D.</dc:creator>
<dc:creator>Adams, P. D.</dc:creator>
<dc:date>2024-05-16</dc:date>
<dc:identifier>doi:10.1101/2024.05.16.594509</dc:identifier>
<dc:title><![CDATA[Likelihood-based interactive local docking into cryo-EM maps in ChimeraX]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.15.594447v1?rss=1">
<title>
<![CDATA[
The mitochondrial stress- induced protein carboxyl-terminal alanine and threonine tailing (msiCAT-tailing) promotes glioblastoma tumorigenesis by modulating mitochondrial functions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.15.594447v1?rss=1</link>
<description><![CDATA[
The rapid and sustained proliferation of cancer cells necessitates increased protein production, which, along with their disrupted metabolism, elevates the likelihood of translation errors. Ribosome-associated quality control (RQC), a recently identified mechanism, mitigates ribosome collisions resulting from frequent translation stalls. However, the precise pathophysiological role of the RQC pathway in oncogenesis remains ambiguous. Our research centered on the pathogenic implications of mitochondrial stress-induced protein carboxyl-terminal alanine and threonine tailing (msiCAT-tailing), a specific RQC response to translational arrest on the outer mitochondrial membrane, in glioblastoma (GBM). The presence of msiCAT-tailed mitochondrial proteins was observed commonly in glioblastoma stem cells (GSCs). The exogenous introduction of the mitochondrial ATP synthase F1 subunit alpha (ATP5) protein, accompanied by artificial CAT-tail mimicking sequences, enhanced mitochondrial membrane potential ({Delta}{Psi}m) and inhibited the formation of the mitochondrial permeability transition pore (MPTP). These alterations in mitochondrial characteristics provided resistance to staurosporine (STS)-induced apoptosis in GBM cells. Consequently, msiCAT-tailing can foster cell survival and migration, whereas blocking msiCAT-tailing via genetic or pharmacological intervention can impede GBM cell overgrowth.

Impact StatementThe Carboxyl-Terminal Alanine-Threonine-tailed protein ATP5 helps glioblastoma mitochondria maintain a high membrane potential and keep the permeability transition pore closed, thereby promoting tumor growth and increasing resistance to apoptosis.

HighlightsO_LIGlioblastoma (GBM) cells have a disturbed RQC pathway
C_LIO_LImsiCAT-tailing on ATP5 in GBM cells increases mitochondrial membrane potential
C_LIO_LIThis msiCAT-tailing prevents MPTP opening
C_LIO_LIATP5 msiCAT-tailing also inhibits drug-induced apoptosis in GBM cells
C_LIO_LIBlocking msiCAT-tailing impedes the overall growth of GBM cells
C_LI
]]></description>
<dc:creator>Cai, T.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Reddy, E.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Tang, Y.</dc:creator>
<dc:creator>Mondal, I.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Ho, W. S.</dc:creator>
<dc:creator>Lu, R. O.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:date>2024-05-18</dc:date>
<dc:identifier>doi:10.1101/2024.05.15.594447</dc:identifier>
<dc:title><![CDATA[The mitochondrial stress- induced protein carboxyl-terminal alanine and threonine tailing (msiCAT-tailing) promotes glioblastoma tumorigenesis by modulating mitochondrial functions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.19.594881v1?rss=1">
<title>
<![CDATA[
Transcriptional control of central T cell tolerance by NR4A family nuclear receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.19.594881v1?rss=1</link>
<description><![CDATA[
Although deletion of self-reactive thymocytes and their diversion into regulatory T cell (Treg) lineage are critical for immune tolerance and homeostasis, the molecular pathways that link antigen recognition to these fates are incompletely understood. The Nr4a nuclear hormone receptors are transcriptionally upregulated in response to TCR signaling in the thymus and are implicated in both deletion and diversion, but the mechanisms by which they operate are not clear. Redundancy among the family members and their requirement for Treg generation and maintenance have obscured their role in negative selection. Here we take advantage of competitive bone marrow chimeras and the OT-II/RIPmOVA model to demonstrate that Nr4a1 and Nr4a3 are essential for upregulation of Bcl2l11/BIM and negative selection by tissue-restricted model self-antigen (TRA). Moreover, we reveal that the Nr4a family is absolutely required for full induction of a broad transcriptional program triggered in self-reactive thymocytes by TRA recognition, and conserved across model systems and the natural repertoire. Importantly, both model self antigen-specific TCR Tg and polyclonal thymocytes lacking Nr4a1/3 that escape negative selection acquire an anergy-like program that persists in the periphery and is also evident among wild-type recent thymic emigrants (RTEs). We propose that the Nr4a family transduces TCR signals during thymic development to enforce the fates of highly self-reactive clones, mediating not only deletion and Treg diversion, but also contributing to a cell-intrinsic, persistent anergy-like program that may operate at the margins of canonical thymic tolerance mechanisms to restrain self-reactive T cells after thymic egress.
]]></description>
<dc:creator>Nielsen, H. V.</dc:creator>
<dc:creator>Mueller, J. L.</dc:creator>
<dc:creator>Hiwa, R.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Proekt, I.</dc:creator>
<dc:creator>Rackaityte, E.</dc:creator>
<dc:creator>Aylard, D.</dc:creator>
<dc:creator>Scharer, C. D.</dc:creator>
<dc:creator>Anderson, M.</dc:creator>
<dc:creator>Au-Yeung, B. B.</dc:creator>
<dc:creator>Zikherman, J.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.19.594881</dc:identifier>
<dc:title><![CDATA[Transcriptional control of central T cell tolerance by NR4A family nuclear receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.20.595039v1?rss=1">
<title>
<![CDATA[
Histologic variants in bladder cancer harbor aggressive molecular features including TM4SF1 expression and a CA125+ cell state 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.20.595039v1?rss=1</link>
<description><![CDATA[
Histologic variant (HV) subtypes of bladder cancer are clinically aggressive tumors that are more resistant to standard therapy compared to conventional urothelial carcinoma (UC). Little is known about the transcriptional programs that account for the morphological and biological differences in HV tumors. To investigate the tumor biology of HV bladder cancers, we generated a single cell RNA sequencing (scRNA- seq) atlas of nine HV tumors and three UC tumors. Our analyses revealed a tumor cell state specific to HVs that is characterized by expression of MUC16 (CA125), KRT24, and WISP2. This CA125+ cell state bears transcriptional hallmarks of epithelial-mesenchymal transition, is enriched in metastases, is predicted to be highly chemotherapy resistant, and is linked with poor survival, suggesting that this cell state plays an important role in the aggressive biology of HV tumors. Our analyses also provide novel evidence of transcriptional "mimicry" between HVs and histologically similar non-urothelial cell types. Lastly, we identified higher expression of TM4SF1, a cell surface protein associated with cancer metastasis, in HV tumor cells compared to UC tumor cells. Finally, CAR T cells engineered against TM4SF1 protein demonstrated in vitro and in vivo activity against bladder cancer cell lines in a TM4SF1 expression- dependent manner, highlighting its potential as a therapeutic target in bladder cancer.

One sentence summarySingle cell RNA sequencing of primary bladder cancers identified a CA125+ cell state specific to histologic variants that is associated with aggressive biological features and TM4SF1 as a novel therapeutic target for histologic variant subtypes of bladder cancer which can be targeted by anti- TM4SF1 CAR T cells.
]]></description>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Song, H.</dc:creator>
<dc:creator>Yip, E.</dc:creator>
<dc:creator>Chang, K.</dc:creator>
<dc:creator>Allegakoen, P.</dc:creator>
<dc:creator>Lu, K. L.</dc:creator>
<dc:creator>Hui, K.</dc:creator>
<dc:creator>Pham, J.</dc:creator>
<dc:creator>Kasap, C.</dc:creator>
<dc:creator>Kumar, V.</dc:creator>
<dc:creator>Gayle, J.</dc:creator>
<dc:creator>Stohr, B. A.</dc:creator>
<dc:creator>Ding, C.-K. C.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:creator>Meng, M. V.</dc:creator>
<dc:creator>Chou, J.</dc:creator>
<dc:creator>Porten, S. P.</dc:creator>
<dc:creator>Huang, F. W.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.20.595039</dc:identifier>
<dc:title><![CDATA[Histologic variants in bladder cancer harbor aggressive molecular features including TM4SF1 expression and a CA125+ cell state]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.20.595060v1?rss=1">
<title>
<![CDATA[
MHConstructor: A high-throughput, haplotype-informed solution to the MHC assembly challenge 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.20.595060v1?rss=1</link>
<description><![CDATA[
The extremely high levels of genetic polymorphism within the human major histocompatibility complex (MHC) limit the usefulness of reference-based alignment methods for sequence assembly. We incorporate a short read de novo assembly algorithm into a workflow for novel application to the MHC. MHConstructor is a containerized pipeline designed for high-throughput, haplotype-informed, reproducible assembly of both whole genome sequencing and target-capture short read data in large, population cohorts. To-date, no other self-contained tool exists for the generation of de novo MHC assemblies from short read data. MHConstructor facilitates wide-spread access to high quality, alignment-free MHC sequence analysis.
]]></description>
<dc:creator>Wade, K. J.</dc:creator>
<dc:creator>Suseno, R.</dc:creator>
<dc:creator>Kizer, K.</dc:creator>
<dc:creator>Williams, J.</dc:creator>
<dc:creator>Boquett, J.</dc:creator>
<dc:creator>Caillier, S.</dc:creator>
<dc:creator>Pollock, N. R.</dc:creator>
<dc:creator>Renschen, A.</dc:creator>
<dc:creator>Santaniello, A.</dc:creator>
<dc:creator>Oksenberg, J. R.</dc:creator>
<dc:creator>Norman, P. J.</dc:creator>
<dc:creator>Augusto, D. G.</dc:creator>
<dc:creator>Hollenbach, J. A.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.20.595060</dc:identifier>
<dc:title><![CDATA[MHConstructor: A high-throughput, haplotype-informed solution to the MHC assembly challenge]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.21.595212v1?rss=1">
<title>
<![CDATA[
A role for pH dynamics regulating transcription factor DNA binding selectivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.21.595212v1?rss=1</link>
<description><![CDATA[
Intracellular pH (pHi) dynamics regulates diverse cell processes such as proliferation, dysplasia, and differentiation, often mediated by the protonation state of a functionally critical histidine residue in endogenous pH sensing proteins. How pHi dynamics can directly regulate gene expression and whether transcription factors can function as pH sensors has received limited attention. We tested the prediction that transcription factors with a histidine in their DNA binding domain (DBD) that forms hydrogen bonds with nucleotides can have pH-regulated activity, which is relevant to more than 85 transcription factors in distinct families, including FOX, KLF, SOX and MITF/Myc. Focusing on FOX family transcription factors, we used unbiased SELEX-seq to identify pH-dependent DNA binding motif preferences, then confirm pH-regulated binding affinities for FOXC2, FOXM1, and FOXN1 to a canonical FkhP DNA motif that are 2.5 to 7.5 greater at pH 7.0 compared with pH 7.5. For FOXC2, we also find greater activity for an FkhP motif at lower pHi in cells and that pH-regulated binding and activity are dependent on a conserved histidine (His122) in the DBD. RNA-seq with FOXC2 also reveals pH-dependent differences in enriched promoter motifs. Our findings identify pH-regulated transcription factor-DNA binding selectivity with relevance to how pHi dynamics can regulate gene expression for myriad cell behaviours.
]]></description>
<dc:creator>Kisor, K. P.</dc:creator>
<dc:creator>Ruiz, D. G.</dc:creator>
<dc:creator>Jacobson, M. P.</dc:creator>
<dc:creator>Barber, D. L.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.21.595212</dc:identifier>
<dc:title><![CDATA[A role for pH dynamics regulating transcription factor DNA binding selectivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.21.595135v1?rss=1">
<title>
<![CDATA[
Norepinephrine Signals Through Astrocytes To Modulate Synapses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.21.595135v1?rss=1</link>
<description><![CDATA[
AbstractLocus coeruleus (LC)-derived norepinephrine (NE) drives network and behavioral adaptations to environmental saliencies by reconfiguring circuit connectivity, but the underlying synapse-level mechanisms are elusive. Here, we show that NE remodeling of synaptic function is independent from its binding on neuronal receptors. Instead, astrocytic adrenergic receptors and Ca2+ dynamics fully gate the effect of NE on synapses as the astrocyte-specific deletion of adrenergic receptors and three independent astrocyte-silencing approaches all render synapses insensitive to NE. Additionally, we find that NE suppression of synaptic strength results from an ATP-derived and adenosine A1 receptor-mediated control of presynaptic efficacy. An accompanying study from Chen et al. reveals the existence of an analogous pathway in the larval zebrafish and highlights its importance to behavioral state transitions. Together, these findings fuel a new model wherein astrocytes are a core component of neuromodulatory systems and the circuit effector through which norepinephrine produces network and behavioral adaptations, challenging an 80-year-old status quo.
]]></description>
<dc:creator>Lefton, K. B.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Yen, A.</dc:creator>
<dc:creator>Okuda, T.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Dai, Y.</dc:creator>
<dc:creator>Walsh, S.</dc:creator>
<dc:creator>Manno, R.</dc:creator>
<dc:creator>Dougherty, J.</dc:creator>
<dc:creator>Samineni, V. K.</dc:creator>
<dc:creator>Simpson, P. C.</dc:creator>
<dc:creator>Papouin, T.</dc:creator>
<dc:date>2024-05-22</dc:date>
<dc:identifier>doi:10.1101/2024.05.21.595135</dc:identifier>
<dc:title><![CDATA[Norepinephrine Signals Through Astrocytes To Modulate Synapses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.21.595187v1?rss=1">
<title>
<![CDATA[
Analyzing high-throughput assay data to advance the rapid screening of environmental chemicals for human reproductive toxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.21.595187v1?rss=1</link>
<description><![CDATA[
While high-throughput (HTP) assays have been proposed as platforms to rapidly assess reproductive toxicity, there is currently a lack of established assays that specifically address germline development/function and fertility. We assessed the applicability domains of yeast (S. cerevisiae) and nematode (C. elegans) HTP assays in toxicity screening of 124 environmental chemicals, determining their agreement in identifying toxicants and their concordance with reproductive toxicity in vivo. We integrated data generated in the two models and compared results using a streamlined, semi-automated benchmark dose (BMD) modeling approach. We then extracted and modeled relevant mammalian in vivo data available for the matching chemicals included in the Toxicological Reference Database (ToxRefDB). We ranked potencies of common compounds using the BMD and evaluated correlation between the datasets using Pearson and Spearman correlation coefficients. We found moderate to good correlation across the three data sets, with r = 0.48 (95% CI: 0.28-1.00, p<0.001) and rs = 0.40 (p=0.002) for the parametric and rank order correlations between the HTP BMDs; r = 0.95 (95% CI: 0.76-1.00, p=0.0005) and rs = 0.89 (p=0.006) between the yeast assay and ToxRefDB BMDs; and r = 0.81 (95% CI: 0.28-1.00, p=0.014) and rs = 0.75 (p=0.033) between the worm assay and ToxRefDB BMDs. Our findings underscore the potential of these HTP assays to identify environmental chemicals that exhibit reproductive toxicity. Integrating these HTP datasets into mammalian in vivo prediction models using machine learning methods could further enhance the predictive value of these assays in future rapid screening efforts.
]]></description>
<dc:creator>Varshavsky, J. R.</dc:creator>
<dc:creator>Lam, J.</dc:creator>
<dc:creator>Cooper, C.</dc:creator>
<dc:creator>Allard, P.</dc:creator>
<dc:creator>Fung, J.</dc:creator>
<dc:creator>Oke, A.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Robinson, J.</dc:creator>
<dc:creator>Woodruff, T.</dc:creator>
<dc:date>2024-05-22</dc:date>
<dc:identifier>doi:10.1101/2024.05.21.595187</dc:identifier>
<dc:title><![CDATA[Analyzing high-throughput assay data to advance the rapid screening of environmental chemicals for human reproductive toxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.23.595237v1?rss=1">
<title>
<![CDATA[
Human neural dynamics of real-world and imagined navigation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.23.595237v1?rss=1</link>
<description><![CDATA[
The ability to form episodic memories and later imagine them is integral to the human experience, influencing our recollection of the past and our ability to envision the future. While research on spatial navigation in rodents suggests the involvement of the medial temporal lobe (MTL), especially the hippocampus, in these cognitive functions, it is uncertain if these insights apply to the human MTL, especially regarding imagination and the reliving of events. Importantly, by involving human participants, imaginations can be explicitly instructed and their mental experiences verbally reported. In this study, we investigated the role of hippocampal theta oscillations in both real-world and imagined navigation, leveraging motion capture and intracranial electroencephalographic recordings from individuals with chronically implanted MTL electrodes who could move freely. Our results revealed intermittent theta dynamics, particularly within the hippocampus, which encoded spatial geometry and partitioned navigational routes into linear segments during real-world navigation. During imagined navigation, theta dynamics exhibited similar, repetitive patterns despite the absence of external environmental cues. Furthermore, a computational model, generalizing from real-world to imagined navigation, successfully reconstructed participants imagined positions using neural data. These findings offer unique insights into the neural mechanisms underlying human navigation and imagination, with implications for understanding episodic memory formation and retrieval in real-world settings.
]]></description>
<dc:creator>Seeber, M.</dc:creator>
<dc:creator>Stangl, M.</dc:creator>
<dc:creator>Vallejo, M.</dc:creator>
<dc:creator>Topalovic, U.</dc:creator>
<dc:creator>Hiller, S.</dc:creator>
<dc:creator>Halpern, C. H.</dc:creator>
<dc:creator>Langevin, J.-P.</dc:creator>
<dc:creator>Rao, V. R.</dc:creator>
<dc:creator>Fried, I.</dc:creator>
<dc:creator>Eliashiv, D.</dc:creator>
<dc:creator>Suthana, N.</dc:creator>
<dc:date>2024-05-23</dc:date>
<dc:identifier>doi:10.1101/2024.05.23.595237</dc:identifier>
<dc:title><![CDATA[Human neural dynamics of real-world and imagined navigation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.24.595367v1?rss=1">
<title>
<![CDATA[
To be unique or blend in: dynamics of male African elephant character durability across time and social contexts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.24.595367v1?rss=1</link>
<description><![CDATA[
Post-dispersal male African elephants (Loxodonta africana) live within complex social networks. To quantify the durability of male elephant character (or personality) within these networks, we employed behavioral repeatability analysis tools across social and environmental contexts. We collected behavioral data from thirty-four individually-identified male elephants at the same waterhole over five field seasons (2007-2011) in Etosha National Park, Namibia. Using repeatability models to assess ten behavioral categories, we found five behaviors (affiliation, aggression, dominance, self-directed anxious, and self-directed comfort) were consistent at the individual level. Interestingly, some of these behaviors were also significantly repeatable, depending on social context. In particular, the presence of younger males and a keystone male (i.e., the most dominant and socially-integrated individual during our study period) had the biggest impact on adult male behaviors. Surprisingly, the presence of elephants in musth had little impact. Finally, we found that younger individuals were more alike in their overall character profiles than older males, further supporting the hypothesis that male elephants develop unique, yet socially-flexible character types as they age. These results demonstrate that male elephants possess distinct character traits that are also behaviorally adaptable, depending on the social context. Overall, our research further uncovers the complexity of male elephant individuality and social dynamics that can be leveraged to improve in-situ and ex-situ management and conservation decisions for the species.
]]></description>
<dc:creator>O'Connell-Rodwell, C. E.</dc:creator>
<dc:creator>Berezin, J. L.</dc:creator>
<dc:creator>Kinzley, C.</dc:creator>
<dc:creator>Freeman, P. T.</dc:creator>
<dc:creator>Sandri, M. N.</dc:creator>
<dc:creator>Kieschnick, D.</dc:creator>
<dc:creator>Abarca, M.</dc:creator>
<dc:creator>Hayssen, V.</dc:creator>
<dc:date>2024-05-26</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.595367</dc:identifier>
<dc:title><![CDATA[To be unique or blend in: dynamics of male African elephant character durability across time and social contexts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.23.595575v1?rss=1">
<title>
<![CDATA[
Loss of neutrophil Shp1 produces hemorrhagic and lethal acute lung injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.23.595575v1?rss=1</link>
<description><![CDATA[
The acute respiratory distress syndrome (ARDS) is associated with significant morbidity and mortality and neutrophils are critical to its pathogenesis. Neutrophil activation is closely regulated by inhibitory tyrosine phosphatases including Src homology region 2 domain containing phosphatase-1 (Shp1). Here, we report that loss of neutrophil Shp1 in mice produced hyperinflammation and lethal pulmonary hemorrhage in sterile inflammation and pathogen-induced models of acute lung injury (ALI) through a Syk kinase-dependent mechanism. We observed large intravascular neutrophil clusters, perivascular inflammation, and excessive neutrophil extracellular traps in neutrophil-specific Shp1 knockout mice suggesting an underlying mechanism for the observed pulmonary hemorrhage. Targeted immunomodulation through the administration of a Shp1 activator (SC43) reduced agonist-induced reactive oxygen species in vitro and ameliorated ALI-induced alveolar neutrophilia and NETs in vivo. We propose that the pharmacologic activation of Shp1 has the potential to fine-tune neutrophil hyperinflammation that is central to the pathogenesis of ARDS.
]]></description>
<dc:creator>Moussavi-Harami, S. F.</dc:creator>
<dc:creator>Cleary, S. J.</dc:creator>
<dc:creator>Magnen, M.</dc:creator>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>Conrad, C.</dc:creator>
<dc:creator>English, B. C.</dc:creator>
<dc:creator>Qiu, L.</dc:creator>
<dc:creator>Wang, K. M.</dc:creator>
<dc:creator>Abram, C. L.</dc:creator>
<dc:creator>Lowell, C. A.</dc:creator>
<dc:creator>Looney, M. R.</dc:creator>
<dc:date>2024-05-28</dc:date>
<dc:identifier>doi:10.1101/2024.05.23.595575</dc:identifier>
<dc:title><![CDATA[Loss of neutrophil Shp1 produces hemorrhagic and lethal acute lung injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.24.595727v1?rss=1">
<title>
<![CDATA[
Decellularized biohybrid nerve promotes motor axon projections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.24.595727v1?rss=1</link>
<description><![CDATA[
Developing nerve grafts with intact mesostructures, superior conductivity, minimal immunogenicity, and improved tissue integration is essential for the treatment and restoration of neurological dysfunctions. A key factor is promoting directed axon growth into the grafts. To achieve this, we developed biohybrid nerves using decellularized rat sciatic nerve modified by in situ polymerization of poly(3,4-ethylenedioxythiophene) (PEDOT). We compared nine biohybrid nerves with varying polymerization conditions and cycles, selecting the best candidate through material characterization. Our results showed that a 1:1 ratio of FeCl3 oxidant to ethylenedioxythiophene (EDOT) monomer, cycled twice, provided superior conductivity (>0.2 mS/cm), mechanical alignment, intact mesostructures, and high compatibility with cells and blood. To test the biohybrid nerves effectiveness in promoting motor axon growth, we used human Spinal Cord Spheroids (hSCSs) from HUES 3 Hb9:GFP cells, with motor axons labeled with green fluorescent protein (GFP). Seeding hSCS onto one end of the conduit allowed motor axon outgrowth into the biohybrid nerve. Our construct effectively promoted directed motor axon growth, which improved significantly after seeding the grafts with Schwann cells. This study presents a promising approach for reconstructing axonal tracts in humans.
]]></description>
<dc:creator>Mehta, A. S.</dc:creator>
<dc:creator>Zhang, S. L.</dc:creator>
<dc:creator>Xie, X.</dc:creator>
<dc:creator>Khanna, S.</dc:creator>
<dc:creator>Tropp, J.</dc:creator>
<dc:creator>Ji, X.</dc:creator>
<dc:creator>Daso, R.</dc:creator>
<dc:creator>Franz, C.</dc:creator>
<dc:creator>Jordan, S.</dc:creator>
<dc:creator>Rivnay, J.</dc:creator>
<dc:date>2024-05-29</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.595727</dc:identifier>
<dc:title><![CDATA[Decellularized biohybrid nerve promotes motor axon projections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.28.593642v1?rss=1">
<title>
<![CDATA[
Autism gene variants disrupt enteric neuron migration and cause gastrointestinal dysmotility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.28.593642v1?rss=1</link>
<description><![CDATA[
The comorbidity of autism spectrum disorders and severe gastrointestinal symptoms is well-established, yet the molecular underpinnings remain unknown. The identification of high-confidence large-effect autism risk genes offers the opportunity to identify convergent, underlying biology by studying these genes in the context of the gastrointestinal system. Here we show that the expression of these genes is enriched in human prenatal gut neurons as well as their migratory progenitors, suggesting that the development and/or function of these neurons may be disrupted by autism-associated pathogenic variants, leading to gastrointestinal dysfunction. Here we document the prevalence of gastrointestinal issues in patients with large-effect variants in sixteen of these genes, highlighting dysmotility, consistent with potential enteric neuron dysfunction. Using the high-throughput diploid frog Xenopus tropicalis, we individually target five of these genes (SYNGAP1, CHD8, SCN2A, CHD2, and DYRK1A) and observe disrupted enteric neuronal progenitor migration for each. More extensive analysis of DYRK1A reveals that perturbation causes gut dysmotility in vivo, which can be ameliorated by treatment with a selective serotonin reuptake inhibitor (escitalopram) or a serotonin receptor 6 agonist, identified by in vivo drug screening. This work suggests that atypical development of enteric neurons contributes to the gastrointestinal distress commonly seen in individuals with autism and that increasing serotonin signaling may be a productive therapeutic avenue.
]]></description>
<dc:creator>McCluskey, K. E.</dc:creator>
<dc:creator>Stovell, K. M.</dc:creator>
<dc:creator>Law, K.</dc:creator>
<dc:creator>Kostyanovskaya, E.</dc:creator>
<dc:creator>Schmidt, J.</dc:creator>
<dc:creator>Exner, C. R.</dc:creator>
<dc:creator>Dea, J.</dc:creator>
<dc:creator>Brimble, E.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Willsey, A. J.</dc:creator>
<dc:creator>Willsey, H. R.</dc:creator>
<dc:date>2024-05-29</dc:date>
<dc:identifier>doi:10.1101/2024.05.28.593642</dc:identifier>
<dc:title><![CDATA[Autism gene variants disrupt enteric neuron migration and cause gastrointestinal dysmotility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.29.596373v1?rss=1">
<title>
<![CDATA[
Predicted mechanistic impacts of human protein missense variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.29.596373v1?rss=1</link>
<description><![CDATA[
Genome sequencing efforts have led to the discovery of tens of millions of protein missense variants found in the human population with the majority of these having no annotated role and some likely contributing to trait variation and disease. Sequence-based artificial intelligence approaches have become highly accurate at predicting variants that are detrimental to the function of proteins but they do not inform on mechanisms of disruption. Here we combined sequence and structure-based methods to perform proteome-wide prediction of deleterious variants with information on their impact on protein stability, protein-protein interactions and small-molecule binding pockets. AlphaFold2 structures were used to predict approximately 100,000 small-molecule binding pockets and stability changes for over 200 million variants. To inform on protein-protein interfaces we used AlphaFold2 to predict structures for nearly 500,000 protein complexes. We illustrate the value of mechanism-aware variant effect predictions to study the relation between protein stability and abundance and the structural properties of interfaces underlying trans protein quantitative trait loci (pQTLs). We characterised the distribution of mechanistic impacts of protein variants found in patients and experimentally studied example disease linked variants in FGFR1.
]]></description>
<dc:creator>Janes, J.</dc:creator>
<dc:creator>Muller, M.</dc:creator>
<dc:creator>Selvaraj, S.</dc:creator>
<dc:creator>Manoel, D.</dc:creator>
<dc:creator>Stephenson, J.</dc:creator>
<dc:creator>Goncalves, C.</dc:creator>
<dc:creator>Lafita, A.</dc:creator>
<dc:creator>Polacco, B.</dc:creator>
<dc:creator>Obernier, K.</dc:creator>
<dc:creator>Alasoo, K.</dc:creator>
<dc:creator>Lemos, M. C.</dc:creator>
<dc:creator>Krogan, N.</dc:creator>
<dc:creator>Martin, M.</dc:creator>
<dc:creator>Saraiva, L. R.</dc:creator>
<dc:creator>Burke, D.</dc:creator>
<dc:creator>Beltrao, P.</dc:creator>
<dc:date>2024-05-29</dc:date>
<dc:identifier>doi:10.1101/2024.05.29.596373</dc:identifier>
<dc:title><![CDATA[Predicted mechanistic impacts of human protein missense variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.28.596151v1?rss=1">
<title>
<![CDATA[
Chromatin profiling identifies putative dual roles for H3K27me3 in regulating transposons and cell type specific genes in choanoflagellates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.28.596151v1?rss=1</link>
<description><![CDATA[
Gene expression is tightly controlled during animal development to allow the formation of specialized cell types. Our understanding of how animals evolved this exquisite regulatory control remains elusive, but evidence suggests that changes in chromatin-based mechanisms may have contributed. To investigate this possibility, here we examine chromatin-based gene regulatory features in the closest relatives of animals, choanoflagellates. Using Salpingoeca rosetta as a model system, we examined chromatin accessibility and histone modifications at the genome scale and compared these features to gene expression. We first observed that accessible regions of chromatin are primarily associated with gene promoters and found no evidence of distal gene regulatory elements resembling the enhancers that animals deploy to regulate developmental gene expression. Remarkably, a histone modification deposited by polycomb repressive complex 2, histone H3 lysine 27 trimethylation (H3K27me3), appeared to function similarly in S. rosetta to its role in animals, because this modification decorated genes with cell type-specific expression. Additionally, H3K27me3 marked transposons, retaining what appears to be an ancestral role in regulating these elements. We further uncovered a putative new bivalent chromatin state at cell type-specific genes that consists of H3K27me3 and histone H3 lysine 4 mono-methylation (H3K4me1). Together, our discoveries support the scenario that gene-associated histone modification states that underpin development emerged before the evolution of animal multicellularity.
]]></description>
<dc:creator>Gahan, J. M.</dc:creator>
<dc:creator>Helfrich, L. W.</dc:creator>
<dc:creator>Wetzel, L. A.</dc:creator>
<dc:creator>Bhanu, N. V.</dc:creator>
<dc:creator>Yuan, Z.-F.</dc:creator>
<dc:creator>Garcia, B. A.</dc:creator>
<dc:creator>Klose, R.</dc:creator>
<dc:creator>Booth, D. S.</dc:creator>
<dc:date>2024-05-30</dc:date>
<dc:identifier>doi:10.1101/2024.05.28.596151</dc:identifier>
<dc:title><![CDATA[Chromatin profiling identifies putative dual roles for H3K27me3 in regulating transposons and cell type specific genes in choanoflagellates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.29.596548v1?rss=1">
<title>
<![CDATA[
Dual Mechanism of Action: Exosomes from Human iPSC-Cardiomyocytes and Mesenchymal Stem Cells Restore Injured Myocardium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.29.596548v1?rss=1</link>
<description><![CDATA[
BackgroundTransplantation of mesenchymal stem cells or induced pluripotent stem cell derived cardiomyocytes improve heart function after myocardial infarction in pre-clinical models. Exosomes are extracellular vesicles, 30-150nm in size, which regulate the paracrine signal of the stem cells. We investigated the functional outcomes and biological effects of exosomes from pure populations of human bone marrow derived mesenchymal stem cells (MSCs) and induced pluripotent stem cell derived cardiomyocytes (iCMs) in a porcine acute myocardial infarction model.

MethodsYorkshire swine were subject to proximal left anterior descending artery occlusion with a catheter balloon for 1 hour for ischemia-reperfusion injury. Ten 500ul injections containing 5 x 1011 exosomes isolated from the tissue culture media of iCMs or MSCs were delivered transendocardially into the peri-infarct region. Cardiac function was assessed by magnetic resonance imaging (MRI). Multi-omic analyses were performed in the ex vivo swine peri-infarct specimen to delineate the mechanism of action.

ResultsCardiac MRI at weeks 2 and 4 showed significant improvement in heart function in iCM-derived exosomes while MSC-derived exosomes showed a trend towards improvement. A comparative analysis of transcriptomic sequencing of the porcine peri-infarct tissue and Next Generation Sequencing of the exosome cargo confirmed the dual mechanism of action. The marked improvements seen in cardiac function are conferred by miRNA carried by the exosomes, particularly by cardioprotective reduction in metabolism during acute myocardial injury while promoting concurrent cardiomyocyte cell cycle re-entry and proliferation.

ConclusionsSignificant reduction in myocardial metabolism and increase in proliferation signal pathways were found in both exosome treatment groups; however, distinct sets of microRNAs were found to underlie the mechanism of action in each population of exosomes.
]]></description>
<dc:creator>Tzng, E.</dc:creator>
<dc:creator>Bayardo, N. A.</dc:creator>
<dc:creator>Ikeda, G.</dc:creator>
<dc:creator>Takashima, H.</dc:creator>
<dc:creator>Lyons, J.</dc:creator>
<dc:creator>Bennett, M.</dc:creator>
<dc:creator>O'Brien, C. G.</dc:creator>
<dc:creator>Yang, P. C.</dc:creator>
<dc:date>2024-05-30</dc:date>
<dc:identifier>doi:10.1101/2024.05.29.596548</dc:identifier>
<dc:title><![CDATA[Dual Mechanism of Action: Exosomes from Human iPSC-Cardiomyocytes and Mesenchymal Stem Cells Restore Injured Myocardium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.29.595206v1?rss=1">
<title>
<![CDATA[
Targeting complement C3a receptor resolves mitochondrial hyperfusion and subretinal microglial activation in progranulin-deficient frontotemporal dementia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.29.595206v1?rss=1</link>
<description><![CDATA[
Mutations in progranulin (GRN) cause frontotemporal dementia (GRN-FTD) due to deficiency of the pleiotropic protein progranulin. GRN-FTD exhibits diverse pathologies including lysosome dysfunction, lipofuscinosis, microgliosis, and neuroinflammation. Yet, how progranulin loss causes disease remains unresolved. Here, we report that non-invasive retinal imaging of GRN-FTD patients revealed deficits in photoreceptors and the retinal pigment epithelium (RPE) that correlate with cognitive decline. Likewise, Grn-/- mice exhibit early RPE dysfunction, microglial activation, and subsequent photoreceptor loss. Super-resolution live imaging and transcriptomic analyses identified RPE mitochondria as an early driver of retinal dysfunction. Loss of mitochondrial fission protein 1 (MTFP1) in Grn-/- RPE causes mitochondrial hyperfusion and bioenergetic defects, leading to NF-kB-mediated activation of complement C3a-C3a receptor signaling, which drives further mitochondrial hyperfusion and retinal inflammation. C3aR antagonism restores RPE mitochondrial integrity and limits subretinal microglial activation. Our study identifies a previously unrecognized mechanism by which progranulin modulates mitochondrial integrity and complement-mediated neuroinflammation.
]]></description>
<dc:creator>Tan, L. X.</dc:creator>
<dc:creator>Oertel, F. C.</dc:creator>
<dc:creator>Cheng, A.</dc:creator>
<dc:creator>Cobigo, Y.</dc:creator>
<dc:creator>Keihani, A.</dc:creator>
<dc:creator>Bennett, D.</dc:creator>
<dc:creator>Abdelhak, A.</dc:creator>
<dc:creator>Montes, S. C.</dc:creator>
<dc:creator>Chapman, M.</dc:creator>
<dc:creator>Chen, R.</dc:creator>
<dc:creator>Cordano, C.</dc:creator>
<dc:creator>Ward, M.</dc:creator>
<dc:creator>Casaletto, K.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Rosen, H. H.</dc:creator>
<dc:creator>Boxer, A. L.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Green, A.</dc:creator>
<dc:creator>Elahi, F.</dc:creator>
<dc:creator>Lakkaraju, A.</dc:creator>
<dc:date>2024-06-01</dc:date>
<dc:identifier>doi:10.1101/2024.05.29.595206</dc:identifier>
<dc:title><![CDATA[Targeting complement C3a receptor resolves mitochondrial hyperfusion and subretinal microglial activation in progranulin-deficient frontotemporal dementia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.01.596962v1?rss=1">
<title>
<![CDATA[
CRISPRi-based screen of Autism Spectrum Disorder risk genes in microglia uncovers roles of ADNP in microglia endocytosis and uptake of synaptic material 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.01.596962v1?rss=1</link>
<description><![CDATA[
Autism Spectrum Disorders (ASD) are a set of neurodevelopmental disorders with complex biology. The identification of ASD risk genes from exome-wide association studies and de novo variation analyses has enabled mechanistic investigations into how ASD-risk genes alter development. Most functional genomics studies have focused on the role of these genes in neurons and neural progenitor cells. However, roles for ASD risk genes in other cell types are largely uncharacterized. There is evidence from postmortem tissue that microglia, the resident immune cells of the brain, appear activated in ASD. Here, we used CRISPRi-based functional genomics to systematically assess the impact of ASD risk gene knockdown on microglia activation and phagocytosis. We developed an iPSC-derived microglia-neuron coculture system and high-throughput flow cytometry readout for synaptic pruning to enable parallel CRISPRi-based screening of phagocytosis of beads, synaptosomes, and synaptic pruning. Our screen identified ADNP, a high-confidence ASD risk genes, as a modifier of microglial synaptic pruning. We found that microglia with ADNP loss have altered endocytic trafficking, remodeled proteomes, and increased motility in coculture.
]]></description>
<dc:creator>Teter, O. M.</dc:creator>
<dc:creator>Draeger, N. M.</dc:creator>
<dc:creator>Sattler, S. M.</dc:creator>
<dc:creator>McQuade, A.</dc:creator>
<dc:creator>Holmes, B. B.</dc:creator>
<dc:creator>Papakis, V.</dc:creator>
<dc:creator>Leng, K.</dc:creator>
<dc:creator>Boggess, S.</dc:creator>
<dc:creator>Shin, D.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Wells, J.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:date>2024-06-01</dc:date>
<dc:identifier>doi:10.1101/2024.06.01.596962</dc:identifier>
<dc:title><![CDATA[CRISPRi-based screen of Autism Spectrum Disorder risk genes in microglia uncovers roles of ADNP in microglia endocytosis and uptake of synaptic material]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.02.597025v1?rss=1">
<title>
<![CDATA[
Generalized cue reactivity in dopamine neurons after opioids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.02.597025v1?rss=1</link>
<description><![CDATA[
Cue reactivity is the maladaptive neurobiological and behavioral response upon exposure to drug cues and is a major driver of relapse. The leading hypothesis is that dopamine release by addictive drugs represents a persistently positive reward prediction error that causes runaway enhancement of dopamine responses to drug cues, leading to their pathological overvaluation compared to non-drug reward alternatives. However, this hypothesis has not been directly tested. Here we developed Pavlovian and operant procedures to measure firing responses, within the same dopamine neurons, to drug versus natural reward cues, which we found to be similarly enhanced compared to cues predicting natural rewards in drug-naive controls. This enhancement was associated with increased behavioral reactivity to the drug cue, suggesting that dopamine release is still critical to cue reactivity, albeit not as previously hypothesized. These results challenge the prevailing hypothesis of cue reactivity, warranting new models of dopaminergic function in drug addiction, and provide critical insights into the neurobiology of cue reactivity with potential implications for relapse prevention.
]]></description>
<dc:creator>Lehmann, C. M.</dc:creator>
<dc:creator>Miller, N. E.</dc:creator>
<dc:creator>Nair, V. S.</dc:creator>
<dc:creator>Costa, K. M.</dc:creator>
<dc:creator>Schoenbaum, G.</dc:creator>
<dc:creator>Moussawi, K.</dc:creator>
<dc:date>2024-06-02</dc:date>
<dc:identifier>doi:10.1101/2024.06.02.597025</dc:identifier>
<dc:title><![CDATA[Generalized cue reactivity in dopamine neurons after opioids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.01.596949v1?rss=1">
<title>
<![CDATA[
Cross Tissue DNAm Biomarker Prediction using Transfer Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.01.596949v1?rss=1</link>
<description><![CDATA[
DNA methylation (DNAm) is an epigenetic mechanism vital for regulating gene expression and influencing disease states. Developing accurate DNAm biomarkers often requires data from specific tissues, which are sometimes difficult to access. This study explores the use of Transfer Learning (TL) to predict blood DNAm biomarkers using saliva DNAm data, aiming to overcome limitations posed by sample size and tissue accessibility. We developed TL-based algorithms that integrate DNAm data from multiple tissues. These algorithms were evaluated against traditional Lasso regression and direct saliva DNAm estimates. Our results show that TL significantly improves the prediction accuracy of DNAm biomarkers, outperforming traditional methods in 20 out of 26 biomarkers. We further validated our models using independent datasets, demonstrating that TL-derived predictions reflect known biological relationships, such as sex differences in telomere length and the impact of smoking on DNAm biomarkers. Our findings highlight the potential of TL in enhancing DNAm biomarker prediction across tissues, providing a valuable tool for epigenetic research. The developed algorithms and methodologies are accessible to researchers, fostering advancements in personalized medicine and aging research. This study establishes a framework for utilizing TL to bridge the gap between accessible and pertinent tissue data, paving the way for more accurate and versatile DNAm biomarker applications.

ACM Reference FormatKristen M McGreevy, Brian H Chen, Steve Horvath, and Donatello Telesca. 2024. Cross Tissue DNAm Biomarker Prediction using Transfer Learning. 1, 1 (June 2024), 43 pages. https://doi.org/10.1145/nnnnnnn.nnnnnnn
]]></description>
<dc:creator>McGreevy, K. M.</dc:creator>
<dc:creator>Chen, B. H.</dc:creator>
<dc:creator>Horvath, S.</dc:creator>
<dc:creator>Telesca, D.</dc:creator>
<dc:date>2024-06-03</dc:date>
<dc:identifier>doi:10.1101/2024.06.01.596949</dc:identifier>
<dc:title><![CDATA[Cross Tissue DNAm Biomarker Prediction using Transfer Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.01.596978v1?rss=1">
<title>
<![CDATA[
Differences in phenotype between long-lived memory B cells against Plasmodium falciparum merozoite antigens and variant surface antigens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.01.596978v1?rss=1</link>
<description><![CDATA[
Plasmodium falciparum infections elicit strong humoral immune responses to two main groups of antigens expressed by blood-stage parasites: merozoite antigens that are involved in the erythrocyte invasion process and variant surface antigens that mediate endothelial sequestration of infected erythrocytes. Long-lived B cells against both antigen classes can be detected in the circulation for years after exposure, but have not been directly compared. Here, we studied the phenotype of long-lived memory and atypical B cells to merozoite antigens (MSP1 and AMA1) and variant surface antigens (the CIDR1 domain of PfEMP1) in Ugandan adults before and after local reduction of P. falciparum transmission. After a median of 1.7 years without P. falciparum infections, the percentage of antigen-specific activated B cells declined, but long-lived antigen-specific B cells were still detectable in all individuals. The majority of MSP1/AMA1-specific B cells were CD95+CD11c+ memory B cells, which are primed for rapid differentiation into antibody-secreting cells, and FcRL5-T-bet- atypical B cells. On the other hand, most CIDR1-specific B cells were CD95-CD11c- memory B cells. CIDR1-specific B cells were also enriched among a subset of atypical B cells that seem poised for antigen presentation. These results point to differences in how these antigens are recognized or processed by the immune system and how P. falciparum-specific B cells will respond upon re-infection.
]]></description>
<dc:creator>Reyes, R. A.</dc:creator>
<dc:creator>Turner, L.</dc:creator>
<dc:creator>Ssewanyana, I.</dc:creator>
<dc:creator>Jagannathan, P.</dc:creator>
<dc:creator>Feeney, M. E.</dc:creator>
<dc:creator>Lavstsen, T.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:creator>Bol, S.</dc:creator>
<dc:creator>Bunnik, E. M.</dc:creator>
<dc:date>2024-06-03</dc:date>
<dc:identifier>doi:10.1101/2024.06.01.596978</dc:identifier>
<dc:title><![CDATA[Differences in phenotype between long-lived memory B cells against Plasmodium falciparum merozoite antigens and variant surface antigens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.03.597050v1?rss=1">
<title>
<![CDATA[
Defective interferon signaling in the circulating monocytes of type 2 diabetic mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.03.597050v1?rss=1</link>
<description><![CDATA[
Type 2 diabetes mellitus (T2DM) is associated with poor outcome after stroke. Peripheral monocytes play a critical role in the secondary injury and recovery of damaged brain tissue after stroke, but the underlying mechanisms are largely unclear. To investigate transcriptome changes and molecular networks across monocyte subsets in response to T2DM and stroke, we performed single-cell RNA-sequencing (scRNAseq) from peripheral blood mononuclear cells and bulk RNA-sequencing from blood monocytes from four groups of adult mice, consisting of T2DM model db/db and normoglycemic control db/+ mice with or without ischemic stroke. Via scRNAseq we found that T2DM expands the monocyte population at the expense of lymphocytes, which was validated by flow cytometry. Among the monocytes, T2DM also disproportionally increased the inflammatory subsets with Ly6C+ and negative MHC class II expression (MO.6C+II-). Conversely, monocytes from control mice without stroke are enriched with steady-state classical monocyte subset of MO.6C+II+ but with the least percentage of MO.6C+II- subtype. Apart from enhancing inflammation and coagulation, enrichment analysis from both scRNAseq and bulk RNAseq revealed that T2DM specifically suppressed type-1 and type-2 interferon signaling pathways crucial for antigen presentation and the induction of ischemia tolerance. Preconditioning by lipopolysaccharide conferred neuroprotection against ischemic brain injury in db/+ but not in db/db mice and coincided with a lesser induction of brain Interferon-regulatory-factor-3 in the brains of the latter mice. Our results suggest that the increased diversity and altered transcriptome in the monocytes of T2DM mice underlie the worse stroke outcome by exacerbating secondary injury and potentiating stroke-induced immunosuppression.

Significance StatementThe mechanisms involved in the detrimental diabetic effect on stroke are largely unclear. We show here, for the first time, that peripheral monocytes have disproportionally altered the subsets and changed transcriptome under diabetes and/or stroke conditions. Moreover, genes in the IFN-related signaling pathways are suppressed in the diabetic monocytes, which underscores the immunosuppression and impaired ischemic tolerance under the T2DM condition. Our data raise a possibility that malfunctioned monocytes may systemically and focally affect the host, leading to the poor outcome of diabetes in the setting of stroke. The results yield important clues to molecular mechanisms involved in the detrimental diabetic effect on stroke outcome.
]]></description>
<dc:creator>Omodaka, S.</dc:creator>
<dc:creator>Kato, Y.</dc:creator>
<dc:creator>Sato, Y.</dc:creator>
<dc:creator>Falcone-Juengert, J.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Kanoke, A.</dc:creator>
<dc:creator>Eckalbar, W. L.</dc:creator>
<dc:creator>Endo, H.</dc:creator>
<dc:creator>Hsieh, C. L.</dc:creator>
<dc:creator>Aran, D.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:date>2024-06-03</dc:date>
<dc:identifier>doi:10.1101/2024.06.03.597050</dc:identifier>
<dc:title><![CDATA[Defective interferon signaling in the circulating monocytes of type 2 diabetic mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.04.597225v1?rss=1">
<title>
<![CDATA[
Molecular effects of indoor tanning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.04.597225v1?rss=1</link>
<description><![CDATA[
BackgroundTanning bed users have a significantly increased risk of melanoma, but it remains unclear how indoor tanning drives melanomagenesis. Tanning bed radiation is often thought of as a substitute for natural UV radiation despite differences in the maximum doses, UV content, body sites exposed, and patterns of melanoma that arise.

MethodsTo better understand the epidemiologic trends and etiology of melanoma associated with tanning bed use, we described the patterns of melanoma in patients with quantifiable tanning bed usage and performed exome sequencing of 182 melanocytes from normal skin of a subset of these patients.

ResultsTanning bed users were more likely than non-users to have melanoma on body sites with low cumulative levels of sun damage and were more likely to have multiple melanomas. The melanocytes in normal appearing skin from tanning bed users had higher mutation burdens, a higher proportion of melanocytes with pathogenic mutations, and distinct mutational signatures. These differences were most prominent over body sites that experience comparatively less exposure to natural sunlight.

ConclusionsWe conclude that tanning bed radiation induces melanoma by increasing the mutation burden of melanocytes and by mutagenizing a broader field of melanocytes than are typically exposed to natural sunlight. The unique signatures of mutations in skin cells of tanning users may be attributable to the distinct spectra of radiation emitted from solariums.
]]></description>
<dc:creator>Gerami, P.</dc:creator>
<dc:creator>Tandukar, B.</dc:creator>
<dc:creator>Deivendran, D.</dc:creator>
<dc:creator>Olivares, S.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Tang, J.</dc:creator>
<dc:creator>Tan, T.</dc:creator>
<dc:creator>Sharma, H.</dc:creator>
<dc:creator>Bandari, A. K.</dc:creator>
<dc:creator>Cruz-Pacheco, N.</dc:creator>
<dc:creator>Chang, D.</dc:creator>
<dc:creator>Marty, A.</dc:creator>
<dc:creator>Olshen, A.</dc:creator>
<dc:creator>Murad, N. F.</dc:creator>
<dc:creator>Song, J.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Yeh, I.</dc:creator>
<dc:creator>Shain, H.</dc:creator>
<dc:date>2024-06-05</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.597225</dc:identifier>
<dc:title><![CDATA[Molecular effects of indoor tanning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.04.597353v1?rss=1">
<title>
<![CDATA[
At the onset of active whisking, the input layer of barrel cortex exhibits a 24 h window of increased excitability that depends on prior experience. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.04.597353v1?rss=1</link>
<description><![CDATA[
The development of motor control over sensory organs is a critical milestone, enabling active explo-ration and shaping of the sensory environment. Whether the onset of sensory organ motor control directly influences the development of corresponding sensory cortices remains unknown. Here, we confirm and exploit the late onset of whisking behavior in mice to address this question in the somatosensory system. Using ex vivo electrophysiology, we describe a transient increase in the intrinsic excitability of excitatory neurons in layer IV of the barrel cortex, which processes whisker input, immediately following the onset of active whisking around on postnatal days 13 and 14. This increase in neuronal gain is specific to layer IV, independent of changes in synaptic strength, and requires prior sensory experience. Further, these effects are not expressed in inhibitory in-terneurons in barrel cortex. The transient increase in excitability is not evident in layer II/III of barrel cortex or in the visual cortex upon eye opening, suggesting a unique interaction between the development of active sensing and the thalamocortical input layer in the somatosensory iso-cortex. Predictive modeling indicates that, immediately following the onset of active whisking, changes in active membrane conductances alone can reliably distinguish neurons in control but not whisker-deprived hemispheres. Our findings demonstrate an experience-dependent, lamina-specific refinement of neuronal excitability tightly linked to the emergence of active whisking. This tran-sient increase in the gain of the thalamic input layer coincides with a critical period for synaptic plasticity in downstream layers, suggesting a role in cortical maturation and sensory processing.
]]></description>
<dc:creator>Shallow, M. C.</dc:creator>
<dc:creator>Tian, L.</dc:creator>
<dc:creator>Lin, H.</dc:creator>
<dc:creator>Lefton, K. B.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Dougherty, J.</dc:creator>
<dc:creator>Culver, J. P.</dc:creator>
<dc:creator>Lambo, M. E.</dc:creator>
<dc:creator>Hengen, K. B.</dc:creator>
<dc:date>2024-06-05</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.597353</dc:identifier>
<dc:title><![CDATA[At the onset of active whisking, the input layer of barrel cortex exhibits a 24 h window of increased excitability that depends on prior experience.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.04.597450v1?rss=1">
<title>
<![CDATA[
PBX1 and PBX3 transcription factors regulate SHH expression in the Frontonasal Ectodermal Zone through complementary mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.04.597450v1?rss=1</link>
<description><![CDATA[
Sonic hedgehog (SHH) signaling from the frontonasal ectodermal zone (FEZ) is a key regulator of craniofacial morphogenesis. Along with SHH, pre-B-cell leukemia homeobox (PBX) transcription factors regulate midfacial development. PBXs act in the epithelium during fusion of facial primordia, but their specific interactions with SHH have not been fully investigated. We hypothesized that PBX1/3 regulate SHH expression in the FEZ by activating or repressing transcription. The hypothesis was tested by manipulating PBX1/3 expression in chick embryos and profiling epigenomic landscapes at early developmental stages. PBX1/3 expression was perturbed in the chick face beginning at stage 10 (HH10) using RCAS viruses, and the resulting SHH expression was assessed at HH22. Overexpressing PBX1 expanded SHH expression, while overexpressing PBX3 decreased SHH expression. Conversely, reducing PBX1 expression decreased SHH expression, but reducing PBX3 induced ectopic SHH expression. We performed ATAC-seq and mapped binding of PBX1 and PBX3 with ChIP-seq on the FEZ at HH22 to assess direct interactions of PBX1/3 with the SHH locus. These multi-omics approaches uncovered a 400 bp PBX1-enriched element within intron 1 of SHH (chr2:8,173,222-8,173,621). Enhancer activity of this element was demonstrated by electroporation of reporter constructs in ovo and luciferase reporter assays in vitro. When bound by PBX1, this element upregulates transcription, while it downregulates transcription when bound by PBX3. The present study identifies a cis-regulatory element, named SFE1, that interacts with PBX1/3 to modulate SHH expression in the FEZ and establishes that PBX1 and PBX3 play complementary roles in SHH regulation during embryonic development.
]]></description>
<dc:creator>Mok, C. H.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Losa, M.</dc:creator>
<dc:creator>Risolino, M.</dc:creator>
<dc:creator>Selleri, L.</dc:creator>
<dc:creator>Marcucio, R.</dc:creator>
<dc:date>2024-06-05</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.597450</dc:identifier>
<dc:title><![CDATA[PBX1 and PBX3 transcription factors regulate SHH expression in the Frontonasal Ectodermal Zone through complementary mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.04.597471v1?rss=1">
<title>
<![CDATA[
Expansion of a bacterial operon during cancer treatment ameliorates drug toxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.04.597471v1?rss=1</link>
<description><![CDATA[
Dose-limiting toxicities remain a major barrier to drug development and therapy, revealing the limited predictive power of human genetics. Herein, we demonstrate the utility of a more comprehensive approach to studying drug toxicity through longitudinal study of the human gut microbiome during colorectal cancer (CRC) treatment (NCT04054908) coupled to cell culture and mouse experiments. 16S rRNA gene sequencing revealed significant shifts in gut microbial community structure during oral fluoropyrimidine treatment across multiple patient cohorts, in mouse small and large intestinal contents, and in patient-derived ex vivo communities. Metagenomic sequencing revealed marked shifts in pyrimidine-related gene abundance during oral fluoropyrimidine treatment, including enrichment of the preTA operon, which is sufficient for the inactivation of active metabolite 5-fluorouracil (5-FU). preTA+ bacteria depleted 5-FU in gut microbiota grown ex vivo and the mouse distal gut. Germ-free and antibiotic-treated mice experienced increased fluoropyrimidine toxicity, which was rescued by colonization with the mouse gut microbiota, preTA+ E. coli, or preTA-high CRC patient stool. Finally, preTA abundance was negatively associated with fluoropyrimidine toxicity in patients. Together, these data support a causal, clinically relevant interaction between a human gut bacterial operon and the dose-limiting side effects of cancer treatment. Our approach is generalizable to other drugs, including cancer immunotherapies, and provides valuable insights into host-microbiome interactions in the context of disease.

One Sentence Summary: Gut microbial enzymes can be used to predict and prevent anticancer drug toxicity.
]]></description>
<dc:creator>Trepka, K.</dc:creator>
<dc:creator>Kidder, W. A.</dc:creator>
<dc:creator>Kyaw, T. S.</dc:creator>
<dc:creator>Olson, C. A.</dc:creator>
<dc:creator>Upadhyay, V.</dc:creator>
<dc:creator>Noecker, C.</dc:creator>
<dc:creator>Stanfield, D.</dc:creator>
<dc:creator>Steiding, P.</dc:creator>
<dc:creator>Spanogiannopoulos, P.</dc:creator>
<dc:creator>Dumlao, D.</dc:creator>
<dc:creator>Turnbaugh, J. A.</dc:creator>
<dc:creator>Stachler, M. D.</dc:creator>
<dc:creator>Van Blarigan, E. L.</dc:creator>
<dc:creator>Venook, A. P.</dc:creator>
<dc:creator>Atreya, C. E.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:date>2024-06-06</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.597471</dc:identifier>
<dc:title><![CDATA[Expansion of a bacterial operon during cancer treatment ameliorates drug toxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.04.597478v1?rss=1">
<title>
<![CDATA[
Directionality bias is necessary to explain spatiotemporal progression of pathology in mouse models of tauopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.04.597478v1?rss=1</link>
<description><![CDATA[
Mounting evidence implicates trans-synaptic connectome-based spread as a shared mechanism behind different tauopathic conditions, yet also suggests there is divergent spatiotemporal progression between them. A potential parsimonious explanation for this apparent contradiction could be that different conditions incur differential rates and directional biases in tau transmission along fiber tracts. In this meta-analysis we closely examined this hypothesis and quantitatively tested it using spatiotemporal tau pathology patterns from 11 distinct models across 4 experimental studies. For this purpose, we extended a network-based spread model by incorporating net directionality along the connectome. Our data unambiguously supports the directional transmission hypothesis. First, retrograde bias is an unambiguously better predictor of tau progression than anterograde bias. Second, while spread exhibits retrograde character, our best-fitting biophysical models incorporate the mixed effects of both retrograde- and anterograde-directed spread, with notable tau-strain-specific differences. We also found a nontrivial association between directionality bias and tau strain aggressiveness, with more virulent strains exhibiting less retrograde character. Taken together, our study implicates directional transmission bias in tau transmission along fiber tracts as a general feature of tauopathy spread and a strong candidate explanation for the diversity of spatiotemporal tau progression between conditions. This simple and parsimonious mechanism may potentially fill a critical gap in our knowledge of the spatiotemporal ramification of divergent tauopathies.
]]></description>
<dc:creator>Torok, J.</dc:creator>
<dc:creator>Mezias, C.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:date>2024-06-06</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.597478</dc:identifier>
<dc:title><![CDATA[Directionality bias is necessary to explain spatiotemporal progression of pathology in mouse models of tauopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.04.597496v1?rss=1">
<title>
<![CDATA[
The disease-causing tau V337M mutation induces tau hypophosphorylation and perturbs axon morphology pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.04.597496v1?rss=1</link>
<description><![CDATA[
Tau aggregation is a hallmark of several neurodegenerative diseases, including Alzheimers disease and frontotemporal dementia. There are disease-causing variants of the tau-encoding gene, MAPT, and the presence of tau aggregates is highly correlated with disease progression. However, the molecular mechanisms linking pathological tau to neuronal dysfunction are not well understood. This is in part due to an incomplete understanding of the normal functions of tau in development and aging, and how the associated molecular and cellular processes change in the context of causal disease variants of tau. To address these questions in an unbiased manner, we conducted multi-omic characterization of iPSC-derived neurons harboring the MAPT V337M mutation or MAPT knockdown. RNA-seq, ATAC-seq, and phosphoproteomics revealed that both V337M mutation and tau knockdown perturbed levels of transcripts and phosphorylation of proteins related to axonogenesis or axon morphology. When we directly measured axonogenesis, we found that both MAPT V337M and MAPT knockdown caused decreased axon length. Surprisingly, we found that neurons with V337M tau had much lower tau phosphorylation than neurons with WT tau. CRISPR-based screens uncovered regulators of tau phosphorylation in neurons and found that factors involved in axonogenesis modified tau phosphorylation in both MAPT WT and MAPT V337M neurons. Intriguingly, the p38 MAPK pathway specifically modified tau phosphorylation in MAPT V337M neurons. We propose that V337M tau perturbs tau phosphorylation and axon morphology pathways that are relevant to the normal function of tau in development, which could contribute to previously reported cognitive changes in preclinical MAPT variant carriers.
]]></description>
<dc:creator>Mohl, G. A.</dc:creator>
<dc:creator>Dixon, G.</dc:creator>
<dc:creator>Marzette, E.</dc:creator>
<dc:creator>McKetney, J.</dc:creator>
<dc:creator>Samelson, A. J.</dc:creator>
<dc:creator>Pereda Serras, C.</dc:creator>
<dc:creator>Jin, J.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Boggess, S. C.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:date>2024-06-06</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.597496</dc:identifier>
<dc:title><![CDATA[The disease-causing tau V337M mutation induces tau hypophosphorylation and perturbs axon morphology pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.05.597590v1?rss=1">
<title>
<![CDATA[
Longitudinal Associations Between MicroRNAs and Weight in the Diabetes Prevention Program 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.05.597590v1?rss=1</link>
<description><![CDATA[
OBJECTIVECirculating microRNAs show cross-sectional associations with overweight and obesity. Few studies provided data to differentiate between a snapshot perspective on these associations versus how microRNAs characterize prodromal risk from disease pathology and complications. This study assessed longitudinal relationships between circulating microRNAs and weight at multiple time-points in the Diabetes Prevention Program trial.

RESEARCH DESIGN AND METHODSA subset of participants (n=150) from the Diabetes Prevention Program were included. MicroRNAs were measured from banked plasma using a Fireplex Assay. We used generalized linear mixed models to evaluate relationships between microRNAs and changes in weight at baseline, year-1, and year-2. Logistic regression was used to evaluate whether microRNAs at baseline were associated with weight change after 2 years.

RESULTSIn fully adjusted models that included relevant covariates, seven miRs (i.e., miR-126, miR-15a, miR-192, miR-23a, and miR-27a) were statistically associated with weight over 2 years. MiR-197 and miR-320a remained significant after adjustment for multiple comparisons. Baseline levels of let-7f, miR-17, and miR-320c were significantly associated with 3% weight loss after 2 years in fully adjusted models.

DISCUSSIONThis study provided evidence for longitudinal relationships between circulating microRNAs and weight. Because microRNAs characterize the combined effects of genetic determinants and responses to behavioral determinants, they may provide insights about the etiology of overweight and obesity in the context or risk for common, complex diseases. Additional studies are needed to validate the potential genes and biological pathways that might be targeted by these microRNA biomarkers and have mechanistic implications for weight loss and disease prevention.
]]></description>
<dc:creator>Flowers, E.</dc:creator>
<dc:creator>Stroebel, B.</dc:creator>
<dc:creator>Gong, X.</dc:creator>
<dc:creator>Lewis, K. A.</dc:creator>
<dc:creator>Aouizerat, B.</dc:creator>
<dc:creator>Gadgil, M.</dc:creator>
<dc:creator>Kanaya, A. M.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:date>2024-06-07</dc:date>
<dc:identifier>doi:10.1101/2024.06.05.597590</dc:identifier>
<dc:title><![CDATA[Longitudinal Associations Between MicroRNAs and Weight in the Diabetes Prevention Program]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.06.597731v1?rss=1">
<title>
<![CDATA[
Mapping genes for human face shape: exploration of univariate phenotyping strategies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.06.597731v1?rss=1</link>
<description><![CDATA[
Human facial shape, while strongly heritable, involves both genetic and structural complexity, necessitating precise phenotyping for accurate assessment. Common phenotyping strategies include simplifying 3D facial features into univariate traits such as anthropometric measurements (e.g., inter-landmark distances), unsupervised dimensionality reductions (e.g., principal component analysis (PCA) and auto-encoder (AE) approaches), and assessing resemblance to particular facial gestalts (e.g., syndromic facial archetypes). This study provides a comparative assessment of these strategies in genome-wide association studies (GWASs) of 3D facial shape. Specifically, we investigated inter-landmark distances, PCA and AE-derived latent dimensions, and facial resemblance to random, extreme, and syndromic gestalts within a GWAS of 8,426 individuals of recent European ancestry. Inter-landmark distances exhibit the highest SNP-based heritability as estimated via LD score regression, followed by AE dimensions. Conversely, resemblance scores to extreme and syndromic facial gestalts display the lowest heritability, in line with expectations. Notably, the aggregation of multiple GWASs on facial resemblance to random gestalts reveals the highest number of independent genetic loci. This novel, easy-to-implement phenotyping approach holds significant promise for capturing genetically relevant morphological traits derived from complex biomedical imaging datasets, and its applications extend beyond faces. Nevertheless, these different phenotyping strategies capture different genetic influences on craniofacial shape. Thus, it remains valuable to explore these strategies individually and in combination to gain a more comprehensive understanding of the genetic factors underlying craniofacial shape and related traits.

Author SummaryAdvancements linking variation in the human genome to phenotypes have rapidly evolved in recent decades and have revealed that most human traits are influenced by genetic variants to at least some degree. While many traits, such as stature, are straightforward to acquire and investigate, the multivariate and multipartite nature of facial shape makes quantification more challenging. In this study, we compared the impact of different facial phenotyping approaches on gene mapping outcomes. Our findings suggest that the choice of facial phenotyping method has an impact on apparent trait heritability and the ability to detect genetic association signals. These results offer valuable insights into the importance of phenotyping in genetic investigations, especially when dealing with highly complex morphological traits.
]]></description>
<dc:creator>Yuan, M.</dc:creator>
<dc:creator>Goovaerts, S.</dc:creator>
<dc:creator>Vanneste, M.</dc:creator>
<dc:creator>Matthews, H.</dc:creator>
<dc:creator>Hoskens, H.</dc:creator>
<dc:creator>Richmond, S.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Spritz, R. A.</dc:creator>
<dc:creator>Hallgrimsson, B.</dc:creator>
<dc:creator>Walsh, S.</dc:creator>
<dc:creator>Shriver, M. D.</dc:creator>
<dc:creator>Shaffer, J. R.</dc:creator>
<dc:creator>Weinberg, S. M.</dc:creator>
<dc:creator>Peeters, H.</dc:creator>
<dc:creator>Claes, P.</dc:creator>
<dc:date>2024-06-07</dc:date>
<dc:identifier>doi:10.1101/2024.06.06.597731</dc:identifier>
<dc:title><![CDATA[Mapping genes for human face shape: exploration of univariate phenotyping strategies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.04.596966v1?rss=1">
<title>
<![CDATA[
Insula uses overlapping codes for emotion in self and others 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.04.596966v1?rss=1</link>
<description><![CDATA[
In daily life, we must recognize others emotions so we can respond appropriately. This ability may rely, at least in part, on neural responses similar to those associated with our own emotions. We hypothesized that the insula, a cortical region near the junction of the temporal, parietal, and frontal lobes, may play a key role in this process. We recorded local field potential (LFP) activity in human neurosurgical patients performing two tasks, one focused on identifying their own emotional response and one on identifying facial emotional responses in others. We found matching patterns of gamma- and high-gamma band activity for the two tasks in the insula. Three other regions (MTL, ACC, and OFC) clearly encoded both self- and other-emotions, but used orthogonal activity patterns to do so. These results support the hypothesis that the insula plays a particularly important role in mediating between experienced vs. observed emotions.
]]></description>
<dc:creator>Xiao, J.</dc:creator>
<dc:creator>Adkinson, J. A.</dc:creator>
<dc:creator>Allawala, A. B.</dc:creator>
<dc:creator>Banks, G.</dc:creator>
<dc:creator>Bartoli, E.</dc:creator>
<dc:creator>Fan, X.</dc:creator>
<dc:creator>Mocchi, M.</dc:creator>
<dc:creator>Pascuzzi, B.</dc:creator>
<dc:creator>Pulapaka, S.</dc:creator>
<dc:creator>Franch, M. C.</dc:creator>
<dc:creator>Mathew, S. J.</dc:creator>
<dc:creator>Mathura, R. K.</dc:creator>
<dc:creator>Myers, J.</dc:creator>
<dc:creator>Pirtle, V.</dc:creator>
<dc:creator>Provenza, N. R.</dc:creator>
<dc:creator>Shofty, B.</dc:creator>
<dc:creator>Watrous, A. J.</dc:creator>
<dc:creator>Pitkow, X.</dc:creator>
<dc:creator>Goodman, W. K.</dc:creator>
<dc:creator>Pouratian, N.</dc:creator>
<dc:creator>Sheth, S. A.</dc:creator>
<dc:creator>Bijanki, K. R.</dc:creator>
<dc:creator>Hayden, B. Y.</dc:creator>
<dc:date>2024-06-08</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.596966</dc:identifier>
<dc:title><![CDATA[Insula uses overlapping codes for emotion in self and others]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.05.597671v1?rss=1">
<title>
<![CDATA[
Noise induces intercellular Ca2+ signaling waves and the unfolded protein response in the hearing cochlea 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.05.597671v1?rss=1</link>
<description><![CDATA[
Exposure to loud noise is a common cause of acquired hearing loss. Disruption of subcellular calcium homeostasis and downstream stress pathways in the endoplasmic reticulum and mitochondria, including the unfolded protein response, have been implicated in the pathophysiology of noise-induced hearing loss. However, studies on the association between calcium homeostasis and stress pathways has been limited due to limited ability to measure calcium dynamics in mature-hearing, noise-exposed mice. We used a genetically encoded calcium indicator mouse model in which GcAMP is expressed specifically in hair cells or supporting cells under control of Myo15Cre or Sox2Cre, respectively. We performed live calcium imaging and UPR gene expression analysis in 8-week-old mice exposed to levels of noise that cause cochlear synaptopathy (98 db SPL) or permanent hearing loss (106 dB SPL). UPR activation occurred immediately after noise exposure and was noise dose-dependent, with the pro-apoptotic pathway upregulated only after 106 dB noise exposure. Spontaneous calcium transients in hair cells and intercellular calcium waves in supporting cells, which are present in neonatal cochleae, were quiescent in mature-hearing cochleae, but re-activated upon noise exposure. 106 dB noise exposure was associated with more persistent and expansive ICS wave activity. These findings demonstrate a strong and dose-dependent association between noise exposure, UPR activation, and changes in calcium homeostasis in hair cells and supporting cells, suggesting that targeting these pathways may be effective to develop treatments for noise-induced hearing loss.
]]></description>
<dc:creator>Park, Y.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Ismail Mohamad, N.</dc:creator>
<dc:creator>Matthews, I. R.</dc:creator>
<dc:creator>Santra, P.</dc:creator>
<dc:creator>Sherr, E. H.</dc:creator>
<dc:creator>Chan, D. K.</dc:creator>
<dc:date>2024-06-08</dc:date>
<dc:identifier>doi:10.1101/2024.06.05.597671</dc:identifier>
<dc:title><![CDATA[Noise induces intercellular Ca2+ signaling waves and the unfolded protein response in the hearing cochlea]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.07.597858v1?rss=1">
<title>
<![CDATA[
CLCC1 promotes hepatic neutral lipid flux and nuclear pore complex assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.07.597858v1?rss=1</link>
<description><![CDATA[
Imbalances in lipid storage and secretion lead to the accumulation of hepatocyte lipid droplets (LDs) (i.e., hepatic steatosis). Our understanding of the mechanisms that govern the channeling of hepatocyte neutral lipids towards cytosolic LDs or secreted lipoproteins remains incomplete. Here, we performed a series of CRISPR-Cas9 screens under different metabolic states to uncover mechanisms of hepatic neutral lipid flux. Clustering of chemical-genetic interactions identified CLIC-like chloride channel 1 (CLCC1) as a critical regulator of neutral lipid storage and secretion. Loss of CLCC1 resulted in the buildup of large LDs in hepatoma cells and knockout in mice caused liver steatosis. Remarkably, the LDs are in the lumen of the ER and exhibit properties of lipoproteins, indicating a profound shift in neutral lipid flux. Finally, remote homology searches identified a domain in CLCC1 that is homologous to yeast Brl1p and Brr6p, factors that promote the fusion of the inner and outer nuclear envelopes during nuclear pore complex assembly. Loss of CLCC1 lead to extensive nuclear membrane herniations, consistent with impaired nuclear pore complex assembly. Thus, we identify CLCC1 as the human Brl1p/Brr6p homolog and propose that CLCC1-mediated membrane remodeling promotes hepatic neutral lipid flux and nuclear pore complex assembly.
]]></description>
<dc:creator>Mathiowetz, A. J.</dc:creator>
<dc:creator>Meymand, E. S.</dc:creator>
<dc:creator>Deol, K. K.</dc:creator>
<dc:creator>Parlakgul, G.</dc:creator>
<dc:creator>Lange, M.</dc:creator>
<dc:creator>Pang, S. P.</dc:creator>
<dc:creator>Roberts, M. A.</dc:creator>
<dc:creator>Torres, E. F.</dc:creator>
<dc:creator>Jorgens, D. M.</dc:creator>
<dc:creator>Zalpuri, R.</dc:creator>
<dc:creator>Kang, M.</dc:creator>
<dc:creator>Boone, C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Morgens, D. W.</dc:creator>
<dc:creator>Tso, E.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Talukdar, S.</dc:creator>
<dc:creator>Levine, T.</dc:creator>
<dc:creator>Ku, G.</dc:creator>
<dc:creator>Arruda, A. P.</dc:creator>
<dc:creator>Olzmann, J.</dc:creator>
<dc:date>2024-06-08</dc:date>
<dc:identifier>doi:10.1101/2024.06.07.597858</dc:identifier>
<dc:title><![CDATA[CLCC1 promotes hepatic neutral lipid flux and nuclear pore complex assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.07.598027v1?rss=1">
<title>
<![CDATA[
MITOCHONDRIAL ANTIVIRAL PATHWAYS CONTROL ANTI-HIV RESPONSES AND ISCHEMIC STROKE OUTCOMES VIA THE RIG-1 SIGNALING AND INNATE IMMUNITY MECHANISMS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.07.598027v1?rss=1</link>
<description><![CDATA[
Occludin (ocln) is one of the main regulatory cells of the blood-brain barrier (BBB). Ocln silencing resulted in alterations of the gene expression signatures of a variety of genes of the innate immunity system, including IFN-stimulated genes (ISGs) and the antiviral retinoic acid-inducible gene-1 (RIG-1) signaling pathway, which functions as a regulator of the cytoplasmic sensors upstream of the mitochondrial antiviral signaling protein (MAVS). Indeed, we observed dysfunctional mitochondrial bioenergetics, dynamics, and autophagy in our system. Alterations of mitochondrial bioenergetics and innate immune protection translated into worsened ischemic stroke outcomes in EcoHIV-infected ocln deficient mice. Overall, these results allow for a better understanding of the molecular mechanisms of viral infection in the brain and describe a previously unrecognized role of ocln as a key factor in the control of innate immune responses and mitochondrial dynamics, which affect cerebral vascular diseases such as ischemic stroke.
]]></description>
<dc:creator>Torices, S.</dc:creator>
<dc:creator>Moreno, T.</dc:creator>
<dc:creator>Ramaswamy, S.</dc:creator>
<dc:creator>Naranjo, O.</dc:creator>
<dc:creator>Teglas, T.</dc:creator>
<dc:creator>Osborne, O. M.</dc:creator>
<dc:creator>Park, M.</dc:creator>
<dc:creator>Sun, E.</dc:creator>
<dc:creator>Toborek, M.</dc:creator>
<dc:date>2024-06-08</dc:date>
<dc:identifier>doi:10.1101/2024.06.07.598027</dc:identifier>
<dc:title><![CDATA[MITOCHONDRIAL ANTIVIRAL PATHWAYS CONTROL ANTI-HIV RESPONSES AND ISCHEMIC STROKE OUTCOMES VIA THE RIG-1 SIGNALING AND INNATE IMMUNITY MECHANISMS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.06.597815v1?rss=1">
<title>
<![CDATA[
The Capicua C1 Domain is Required for Full Activity of the CIC::DUX4 Fusion Oncoprotein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.06.597815v1?rss=1</link>
<description><![CDATA[
Rearrangements between genes can yield neomorphic fusions that drive oncogenesis. Fusion oncogenes are made up of fractional segments of the partner genes that comprise them, with each partner potentially contributing some of its own function to the nascent fusion oncoprotein. Clinically, fusion oncoproteins driving one diagnostic entity are typically clustered into a single molecular subset and are often treated a similar fashion. However, knowledge of where specific fusion breakpoints occur in partner genes, and the resulting retention of functional domains in the fusion, is an important determinant of fusion oncoprotein activity and may differ between patients. This study investigates this phenomena through the example of CIC::DUX4, a fusion between the transcriptional repressor capicua (CIC) and the double homeobox 4 gene (DUX4), which drives an aggressive subset of undifferentiated round cell sarcoma. Using a harmonized dataset of over 100 patient fusion breakpoints from the literature, we show that most bona fide CIC::DUX4 fusions retain the C1 domain, which is known to contribute to DNA binding by wild type CIC. Mechanistically, deletion or mutation of the C1 domain reduces, but does not eliminate, activation of CIC target genes by CIC::DUX4. We also find that expression of C1-deleted CIC::DUX4 is capable of exerting intermediate transformation-related phenotypes compared with those imparted by full-length CIC::DUX4, but was not sufficient for tumorigenesis in a subcutaneous mouse model. In summary, our results suggest a supercharging role for the C1 domain in the activity of CIC::DUX4.

Significance StatementFusion oncogenes are neomorphic entities comprised of fractional coding sequences from two partner genes that have been inappropriately rearranged. The functional domains contributed by the partner genes shape the function of the resulting fusion. We use CIC::DUX4, a transcription factor fusion that drives an ultra-rare soft tissue sarcoma, to explore how preferential retention of a partner gene domain may influence the activity of the overall fusion. Our results indicate that the capicua C1 domain is retained in most CIC::DUX4 transcripts and is required for full activity of the CIC::DUX4 oncoprotein. We demonstrate that knowledge of where breakpoints occur and the resulting impact this has on the retention of functional domains can teach us about fusion behavior.
]]></description>
<dc:creator>Luck, C.</dc:creator>
<dc:creator>Jacobs, K. A.</dc:creator>
<dc:creator>Okimoto, R. A.</dc:creator>
<dc:date>2024-06-09</dc:date>
<dc:identifier>doi:10.1101/2024.06.06.597815</dc:identifier>
<dc:title><![CDATA[The Capicua C1 Domain is Required for Full Activity of the CIC::DUX4 Fusion Oncoprotein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.07.597982v1?rss=1">
<title>
<![CDATA[
GABA production induced by imipridones is a targetable and imageable metabolic alteration in diffuse midline gliomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.07.597982v1?rss=1</link>
<description><![CDATA[
Diffuse midline gliomas (DMGs) are lethal primary brain tumors in children. The imipridones ONC201 and ONC206 induce mitochondrial dysfunction and have emerged as promising therapies for DMG patients. However, efficacy as monotherapy is limited, identifying a need for strategies that enhance response. Another hurdle is the lack of biomarkers that report on drug-target engagement at an early timepoint after treatment onset. Here, using 1H-magnetic resonance spectroscopy, which is a non-invasive method of quantifying metabolite pool sizes, we show that accumulation of {psi}-aminobutyric acid (GABA) is an early metabolic biomarker that can be detected within a week of ONC206 treatment, when anatomical alterations are absent, in mice bearing orthotopic xenografts. Mechanistically, imipridones activate the mitochondrial protease ClpP and upregulate the stress-responsive transcription factor ATF4. ATF4, in turn, upregulates glutamate decarboxylase, which synthesizes GABA, and downregulates ABAT, which degrades GABA, leading to GABA accumulation in DMG cells and tumors. Functionally, GABA secreted by imipridone-treated cells acts in an autocrine manner via the GABAB receptor to induce expression of superoxide dismutase (SOD1), which mitigates imipridone-induced oxidative stress and, thereby, curbs apoptosis. Importantly, blocking autocrine GABA signaling using the clinical stage GABAB receptor antagonist SGS-742 exacerbates oxidative stress and synergistically induces apoptosis in combination with imipridones in DMG cells and orthotopic tumor xenografts. Collectively, we identify GABA as a unique metabolic adaptation to imipridones that can be leveraged for non-invasive assessment of drug-target engagement and therapy. Clinical translation of our studies has the potential to enable precision metabolic therapy and imaging for DMG patients.

One Sentence SummaryImipridones induce GABA accumulation in diffuse midline gliomas, an effect that can be leveraged for therapy and non-invasive imaging.
]]></description>
<dc:creator>Batsios, G.</dc:creator>
<dc:creator>Udutha, S.</dc:creator>
<dc:creator>Taglang, C.</dc:creator>
<dc:creator>Gillespie, A. M.</dc:creator>
<dc:creator>Lau, B.</dc:creator>
<dc:creator>Ji, S.</dc:creator>
<dc:creator>Phoenix, T.</dc:creator>
<dc:creator>Mueller, S.</dc:creator>
<dc:creator>Venneti, S.</dc:creator>
<dc:creator>Koschmann, C.</dc:creator>
<dc:creator>Viswanath, P.</dc:creator>
<dc:date>2024-06-10</dc:date>
<dc:identifier>doi:10.1101/2024.06.07.597982</dc:identifier>
<dc:title><![CDATA[GABA production induced by imipridones is a targetable and imageable metabolic alteration in diffuse midline gliomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.11.598512v1?rss=1">
<title>
<![CDATA[
Ferroptosis regulates hemolysis in stored murine and human red blood cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.11.598512v1?rss=1</link>
<description><![CDATA[
Red blood cell (RBC) metabolism regulates hemolysis during aging in vivo and in the blood bank. Here, we leveraged a diversity outbred mouse population to map the genetic drivers of fresh/stored RBC metabolism and extravascular hemolysis upon storage and transfusion in 350 mice. We identify the ferrireductase Steap3 as a critical regulator of a ferroptosis-like process of lipid peroxidation. Steap3 polymorphisms were associated with RBC iron content, in vitro hemolysis, and in vivo extravascular hemolysis both in mice and 13,091 blood donors from the Recipient Epidemiology and Donor evaluation Study. Using metabolite Quantitative Trait Loci analyses, we identified a network of gene products (FADS1/2, EPHX2 and LPCAT3) - enriched in donors of African descent - associated with oxylipin metabolism in stored human RBCs and related to Steap3 or its transcriptional regulator, the tumor protein TP53. Genetic variants were associated with lower in vivo hemolysis in thousands of single-unit transfusion recipients.

HighlightsO_LISteap3 regulates lipid peroxidation and extravascular hemolysis in 350 diversity outbred mice
C_LIO_LISteap3 SNPs are linked to RBC iron, hemolysis, vesiculation in 13,091 blood donors
C_LIO_LImQTL analyses of oxylipins identified ferroptosis-related gene products FADS1/2, EPHX2, LPCAT3
C_LIO_LIFerroptosis markers are linked to hemoglobin increments in transfusion recipients
C_LI

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=199 SRC="FIGDIR/small/598512v1_ufig1.gif" ALT="Figure 1">
View larger version (117K):
org.highwire.dtl.DTLVardef@150fec2org.highwire.dtl.DTLVardef@859c43org.highwire.dtl.DTLVardef@1d60156org.highwire.dtl.DTLVardef@f1b91e_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>D'Alessandro, A.</dc:creator>
<dc:creator>Keele, G. R.</dc:creator>
<dc:creator>Hay, A.</dc:creator>
<dc:creator>Nemkov, T.</dc:creator>
<dc:creator>Earley, E. J.</dc:creator>
<dc:creator>Stephenson, D.</dc:creator>
<dc:creator>Vincent, M.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Dzieciatkowska, M.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>Kleinman, S.</dc:creator>
<dc:creator>Spitalnik, S. L.</dc:creator>
<dc:creator>Roubinian, N.</dc:creator>
<dc:creator>Norris, P. J.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>Page, G. P.</dc:creator>
<dc:creator>Stockwell, B. R.</dc:creator>
<dc:creator>Churchill, G. A.</dc:creator>
<dc:creator>Zimring, J. C.</dc:creator>
<dc:date>2024-06-11</dc:date>
<dc:identifier>doi:10.1101/2024.06.11.598512</dc:identifier>
<dc:title><![CDATA[Ferroptosis regulates hemolysis in stored murine and human red blood cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.13.597459v1?rss=1">
<title>
<![CDATA[
Prosapip1 in the dorsal hippocampus mediates synaptic protein composition, long-term potentiation, and spatial memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.13.597459v1?rss=1</link>
<description><![CDATA[
Prosapip1 is a brain-specific protein localized to the postsynaptic density, where it promotes dendritic spine maturation in primary hippocampal neurons. However, nothing is known about the role of Prosapip1 in vivo. To examine this, we utilized the Cre-loxP system to develop a Prosapip1 neuronal knockout mouse. We found that Prosapip1 controls the synaptic localization of its binding partner SPAR, along with PSD-95 and the GluN2B subunit of the NMDA receptor (NMDAR) in the dorsal hippocampus (dHP). We next sought to identify the potential contribution of Prosapip1 to the activity and function of the NMDAR and found that Prosapip1 plays an important role in NMDAR-mediated transmission and long-term potentiation (LTP) in the CA1 region of the dHP. As LTP is the cellular hallmark of learning and memory, we examined the consequences of neuronal knockout of Prosapip1 on dHP-dependent memory. We found that global or dHP-specific neuronal knockout of Prosapip1 caused a deficit in learning and memory whereas developmental, locomotor, and anxiety phenotypes were normal. Taken together, Prosapip1 in the dHP promotes the proper localization of synaptic proteins which, in turn, facilitates LTP driving recognition, social, and spatial learning and memory.
]]></description>
<dc:creator>Hoisington, Z. W.</dc:creator>
<dc:creator>Gangal, H.</dc:creator>
<dc:creator>Phamluong, K.</dc:creator>
<dc:creator>Shukla, C.</dc:creator>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>Moffat, J. J.</dc:creator>
<dc:creator>Homanics, G. E.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2024-06-13</dc:date>
<dc:identifier>doi:10.1101/2024.06.13.597459</dc:identifier>
<dc:title><![CDATA[Prosapip1 in the dorsal hippocampus mediates synaptic protein composition, long-term potentiation, and spatial memory]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.13.598914v1?rss=1">
<title>
<![CDATA[
Microglia target synaptic sites early during excitatory circuit disassembly in neurodegeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.13.598914v1?rss=1</link>
<description><![CDATA[
During development, microglia prune excess synapses to refine neuronal circuits. In neurodegeneration, the role of microglia-mediated synaptic pruning in circuit remodeling and dysfunction is important for developing therapies aimed at modulating microglial function. Here we analyzed the role of microglia in the synapse disassembly of degenerating postsynaptic neurons in the inner retina. After inducing transient intraocular pressure elevation to injure retinal ganglion cells, microglia increase in number, shift to ameboid morphology, and exhibit greater process movement. Furthermore, due to the greater number of microglia, there is increased colocalization of microglia with synaptic components throughout the inner plexiform layer and with excitatory synaptic sites along individual ganglion cell dendrites. Microglia depletion partially restores ganglion cell function, suggesting that microglia activation may be neurotoxic in early neurodegeneration. Our results demonstrate the important role of microglia in synapse disassembly in degenerating circuits, highlighting their recruitment to synaptic sites early after neuronal injury.

HighlightsEarly after transient intraocular pressure elevation: O_LIMicroglia increase in number, complexity, and process movement
C_LIO_LIMicroglia-synaptic contacts increase in the inner plexiform layer
C_LIO_LIMicroglia-synaptic contacts increase on retinal ganglion cell dendrites
C_LIO_LIMicroglia depletion partially restores ganglion cell function
C_LI
]]></description>
<dc:creator>Yu, A.</dc:creator>
<dc:creator>Tan, L. X.</dc:creator>
<dc:creator>Lakkaraju, A.</dc:creator>
<dc:creator>Santina, L. D.</dc:creator>
<dc:creator>Ou, Y.</dc:creator>
<dc:date>2024-06-14</dc:date>
<dc:identifier>doi:10.1101/2024.06.13.598914</dc:identifier>
<dc:title><![CDATA[Microglia target synaptic sites early during excitatory circuit disassembly in neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.14.598695v1?rss=1">
<title>
<![CDATA[
Transcriptomic comparison of in vitro models of the human placenta 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.14.598695v1?rss=1</link>
<description><![CDATA[
Studying the human placenta through in vitro cell culture methods is necessary due to limited access and amenability of human placental tissue to certain experimental methods as well as distinct anatomical and physiological differences between animal and human placentas. Selecting an in vitro culture model of the human placenta is challenging due to representation of different trophoblast cell types with distinct biological roles and limited comparative studies that define key characteristics of these models. Therefore, the aim of this research was to create a comprehensive transcriptomic comparison of common in vitro models of the human placenta compared to bulk placental tissue from the CANDLE and GAPPS cohorts (N=1083). We performed differential gene expression analysis on publicly available RNA sequencing data from 6 common in vitro models of the human placenta (HTR-8/SVneo, BeWo, JEG-3, JAR, Primary Trophoblasts, and Villous Explants) and compared to CANDLE and GAPPS bulk placental tissue or cytotrophoblast, syncytiotrophoblast, and extravillous trophoblast cell types derived from bulk placental tissue. All in vitro placental models had a substantial number of differentially expressed genes (DEGs, FDR<0.01) compared to the CANDLE and GAPPS placentas (Average DEGs=10,873), and the individual trophoblast cell types (Average DEGs=5,346), indicating that there are vast differences in gene expression compared to bulk and cell-type specific human placental tissue. Hierarchical clustering identified 53 gene clusters with distinct expression profiles across placental models, with 22 clusters enriched for specific KEGG pathways, 7 clusters enriched for high-expression placental genes, and 7 clusters enriched for absorption, distribution, metabolism, and excretion genes. In vitro placental models were classified by fetal sex based on expression of Y-chromosome genes that identified HTR-8/SVneo cells as being of female origin, while JEG-3, JAR, and BeWo cells are of male origin. Overall, none of the models were a close approximation of the transcriptome of bulk human placental tissue, highlighting the challenges with model selection. To enable researchers to select appropriate models, we have compiled data on differential gene expression, clustering, and fetal sex into an accessible web application: "Comparative Transcriptomic Placental Model Atlas (CTPMA)" which can be utilized by researchers to make informed decisions about their selection of in vitro placental models.
]]></description>
<dc:creator>Lapehn, S.</dc:creator>
<dc:creator>Nair, S.</dc:creator>
<dc:creator>Firsick, E. J.</dc:creator>
<dc:creator>MacDonald, J.</dc:creator>
<dc:creator>Thoreson, C.</dc:creator>
<dc:creator>Litch, J. A.</dc:creator>
<dc:creator>Bush, N. R.</dc:creator>
<dc:creator>Kadam, L.</dc:creator>
<dc:creator>Girard, S.</dc:creator>
<dc:creator>Myatt, L.</dc:creator>
<dc:creator>Prasad, B.</dc:creator>
<dc:creator>Sathyanarayana, S.</dc:creator>
<dc:creator>Paquette, A. G.</dc:creator>
<dc:date>2024-06-16</dc:date>
<dc:identifier>doi:10.1101/2024.06.14.598695</dc:identifier>
<dc:title><![CDATA[Transcriptomic comparison of in vitro models of the human placenta]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.15.599084v1?rss=1">
<title>
<![CDATA[
Static and Dynamic Cross-Network Functional Connectivity Shows Elevated Entropy in Schizophrenia Patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.15.599084v1?rss=1</link>
<description><![CDATA[
Schizophrenia (SZ) patients exhibit abnormal static and dynamic functional connectivity across various brain domains. We present a novel approach based on static and dynamic inter-network connectivity entropy (ICE), which represents the entropy of a given networks connectivity to all the other brain networks. This novel approach enables the investigation of how connectivity strength is heterogeneously distributed across available targets in both SZ patients and healthy controls. We analyzed fMRI data from 151 schizophrenia patients and demographically matched 160 healthy controls. Our assessment encompassed both static and dynamic ICE, revealing significant differences in the heterogeneity of connectivity levels across available brain networks between SZ patients and healthy controls (HC). These networks are associated with subcortical (SC), auditory (AUD), sensorimotor (SM), visual (VIS), cognitive control (CC), default mode network (DMN) and cerebellar (CB) functional brain domains. Elevated ICE observed in individuals with SZ suggests that patients exhibit significantly higher randomness in the distribution of time-varying connectivity strength across functional regions from each source network, compared to healthy control group. C-means fuzzy clustering analysis of functional ICE correlation matrices revealed that SZ patients exhibit significantly higher occupancy weights in clusters with weak, low-scale functional entropy correlation, while the control group shows greater occupancy weights in clusters with strong, large-scale functional entropy correlation. k-means clustering analysis on time-indexed ICE vectors revealed that cluster with highest ICE have higher occupancy rates in SZ patients whereas clusters characterized by lowest ICE have larger occupancy rates for control group. Furthermore, our dynamic ICE approach revealed that it appears healthy for a brain to primarily circulate through complex, less structured connectivity patterns, with occasional transitions into more focused patterns. However, individuals with SZ seem to struggle with transiently attaining these more focused and structured connectivity patterns. Proposed ICE measure presents a novel framework for gaining deeper insights into understanding mechanisms of healthy and disease brain states and a substantial step forward in the developing advanced methods of diagnostics of mental health conditions.
]]></description>
<dc:creator>Maksymchuk, N.</dc:creator>
<dc:creator>Bustillo, J. R.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>Preda, A.</dc:creator>
<dc:creator>Calhoun, V. D.</dc:creator>
<dc:date>2024-06-17</dc:date>
<dc:identifier>doi:10.1101/2024.06.15.599084</dc:identifier>
<dc:title><![CDATA[Static and Dynamic Cross-Network Functional Connectivity Shows Elevated Entropy in Schizophrenia Patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.18.599357v1?rss=1">
<title>
<![CDATA[
The effect of circulating neutralizing antibodies on the replication of SARS-CoV-2 variants following post-vaccination infections. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.18.599357v1?rss=1</link>
<description><![CDATA[
The impact of pre-existing neutralizing antibodies (NAbs) titers on SARS-CoV-2 viral shedding dynamics in post-vaccination infection (PVI) are not well understood. We characterized viral shedding longitudinally in nasal specimens in relation to baseline (pre/peri-infection) serum neutralizing antibody titers in 125 participants infected with distinct SARS-CoV-2 variants. Among 68 participants who had received vaccinations, we quantified the effect of baseline serum NAb titers on maximum viral RNA titers and on the duration of infectivity. Baseline NAb titers were higher and efficiently targeted a broader range of variants in participants who received one or two monovalent ancestral booster vaccinations compared to those with a full primary vaccine series. In participants with Delta variant infections, baseline NAb titers targeting Delta were negatively correlated with maximum viral RNA copies. Per log10 increase in baseline NAb IC50, maximum viral load was reduced -2.43 (95% confidence interval [CI] -3.76, -1.11) log10 N copies and days of infectious viral shedding were reduced -2.79 [95% CI: -4.99, -0.60] days. By contrast, in those with Omicron infections (BA.1, BA.2, BA.4 or BA.5 lineages) baseline NAb responses against Omicron lineages did not predict viral outcomes. Our results provide robust estimates of the effect of baseline NAbs on the magnitude and duration of nasal viral replication after PVI (albeit with an unclear effect on transmission) and show how immune escape variants efficiently evade these modulating effects.
]]></description>
<dc:creator>Knight, M. A. G.</dc:creator>
<dc:creator>Kelly, J. D.</dc:creator>
<dc:creator>Lu, S.</dc:creator>
<dc:creator>Tassetto, M. A.</dc:creator>
<dc:creator>Goldberg, S. A.</dc:creator>
<dc:creator>Zhang, A.</dc:creator>
<dc:creator>Pineda-Ramirez, J.</dc:creator>
<dc:creator>Anglin, K.</dc:creator>
<dc:creator>Davidson, M.</dc:creator>
<dc:creator>Chen, J. Y.</dc:creator>
<dc:creator>Fortes-Cobby, M.</dc:creator>
<dc:creator>Park, S.</dc:creator>
<dc:creator>Martinez, A.</dc:creator>
<dc:creator>So, M.</dc:creator>
<dc:creator>Donovan, A.</dc:creator>
<dc:creator>Viswanathan, B.</dc:creator>
<dc:creator>Richardson, E. T.</dc:creator>
<dc:creator>McIlwain, D. R.</dc:creator>
<dc:creator>Gaudilliere, B.</dc:creator>
<dc:creator>Rutishauser, R. L.</dc:creator>
<dc:creator>Chenna, A.</dc:creator>
<dc:creator>Petropoulos, C.</dc:creator>
<dc:creator>Wrin, T.</dc:creator>
<dc:creator>Deeks, S.</dc:creator>
<dc:creator>Abedi, G.</dc:creator>
<dc:creator>Saydah, S.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Briggs-Hagen, M.</dc:creator>
<dc:creator>Midgley, C. M.</dc:creator>
<dc:creator>Peluso, M.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:date>2024-06-19</dc:date>
<dc:identifier>doi:10.1101/2024.06.18.599357</dc:identifier>
<dc:title><![CDATA[The effect of circulating neutralizing antibodies on the replication of SARS-CoV-2 variants following post-vaccination infections.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.21.600041v1?rss=1">
<title>
<![CDATA[
ProBASS: a language model with sequence and structural features for predicting the effect of mutations on binding affinity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.21.600041v1?rss=1</link>
<description><![CDATA[
Protein-protein interactions (PPIs) govern virtually all cellular processes. Even a single mutation within PPI can significantly influence overall protein functionality and potentially lead to various types of diseases. To date, numerous approaches have emerged for predicting the change in free energy of binding ({Delta}{Delta}Gbind) resulting from mutations, yet the majority of these methods lack precision. In recent years, protein language models (PLMs) have been developed and shown powerful predictive capabilities by leveraging both sequence and structural data from protein-protein complexes. Yet, PLMs have not been optimized specifically for predicting {Delta}{Delta}Gbind. We developed an approach to predict effects of mutations on PPI binding affinity based on two most advanced protein language models ESM2 and ESM-IF1 that incorporate PPI sequence and structural features, respectively. We used the two models to generate embeddings for each PPI mutant and subsequently fine-tuned our model by training on a large dataset of experimental {Delta}{Delta}Gbind values. Our model, ProBASS (Protein Binding Affinity from Structure and Sequence) achieved a correlation with experimental {Delta}{Delta}Gbind values of 0.83 {+/-} 0.05 for single mutations and 0.69 {+/-} 0.04 for double mutations when model training and testing was done on the same PDB. Moreover, ProBASS exhibited very high correlation (0.81 {+/-} 0.02) between prediction and experiment when training and testing was performed on a dataset containing 2325 single mutations in 132 PPIs. ProBASS surpasses the state-of-the-art methods in correlation with experimental data and could be further trained as more experimental data becomes available. Our results demonstrate that the integration of extensive datasets containing {Delta}{Delta}Gbind values across multiple PPIs to refine the pre-trained PLMs represents a successful approach for achieving a precise and broadly applicable model for {Delta}{Delta}Gbind prediction, greatly facilitating future protein engineering and design studies.
]]></description>
<dc:creator>Gurusinghe, S. N. S.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>DeGrado, W.</dc:creator>
<dc:creator>Shifman, J. M.</dc:creator>
<dc:date>2024-06-26</dc:date>
<dc:identifier>doi:10.1101/2024.06.21.600041</dc:identifier>
<dc:title><![CDATA[ProBASS: a language model with sequence and structural features for predicting the effect of mutations on binding affinity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.21.599604v1?rss=1">
<title>
<![CDATA[
Long-term NMN treatment increases lifespan and healthspan in mice in a sex dependent manner 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.21.599604v1?rss=1</link>
<description><![CDATA[
Nicotinamide adenine dinucleotide (NAD) is essential for many enzymatic reactions, including those involved in energy metabolism, DNA repair and the activity of sirtuins, a family of defensive deacylases. During aging, levels of NAD+ can decrease by up to 50% in some tissues, the repletion of which provides a range of health benefits in both mice and humans. Whether or not the NAD+ precursor nicotinamide mononucleotide (NMN) extends lifespan in mammals is not known. Here we investigate the effect of long-term administration of NMN on the health, cancer burden, frailty and lifespan of male and female mice. Without increasing tumor counts or severity in any tissue, NMN treatment of males and females increased activity, maintained more youthful gene expression patterns, and reduced overall frailty. Reduced frailty with NMN treatment was associated with increases in levels of Anerotruncus colihominis, a gut bacterium associated with lower inflammation in mice and increased longevity in humans. NMN slowed the accumulation of adipose tissue later in life and improved metabolic health in male but not female mice, while in females but not males, NMN increased median lifespan by 8.5%, possible due to sex-specific effects of NMN on NAD+ metabolism. Together, these data show that chronic NMN treatment delays frailty, alters the microbiome, improves male metabolic health, and increases female mouse lifespan, without increasing cancer burden. These results highlight the potential of NAD+ boosters for treating age-related conditions and the importance of using both sexes for interventional lifespan studies.
]]></description>
<dc:creator>Kane, A. E.</dc:creator>
<dc:creator>Chellappa, K.</dc:creator>
<dc:creator>Schultz, M. B.</dc:creator>
<dc:creator>Arnold, M.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Amorim, J.</dc:creator>
<dc:creator>Diener, C.</dc:creator>
<dc:creator>Zhu, D.</dc:creator>
<dc:creator>Mitchell, S. J.</dc:creator>
<dc:creator>Griffin, P. T.</dc:creator>
<dc:creator>Tian, X.</dc:creator>
<dc:creator>Petty, C.</dc:creator>
<dc:creator>Conway, R.</dc:creator>
<dc:creator>Walsh, K.</dc:creator>
<dc:creator>Shelerud, L.</dc:creator>
<dc:creator>Duesing, C.</dc:creator>
<dc:creator>Mueller, A.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>McNamara, M.</dc:creator>
<dc:creator>Shima, R. T.</dc:creator>
<dc:creator>deCabo, R.</dc:creator>
<dc:creator>Gibbons, S. M.</dc:creator>
<dc:creator>Wu, L. E.</dc:creator>
<dc:creator>Ikeno, Y.</dc:creator>
<dc:creator>Baur, J. A.</dc:creator>
<dc:creator>Rajman, L.</dc:creator>
<dc:creator>Sinclair, D. A.</dc:creator>
<dc:date>2024-06-27</dc:date>
<dc:identifier>doi:10.1101/2024.06.21.599604</dc:identifier>
<dc:title><![CDATA[Long-term NMN treatment increases lifespan and healthspan in mice in a sex dependent manner]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.22.600201v1?rss=1">
<title>
<![CDATA[
Coordinated Regulation of Renal Glucose Reabsorption and Gluconeogenesis by mTORC2 and Potassium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.22.600201v1?rss=1</link>
<description><![CDATA[
BackgroundThe kidney proximal tubule is uniquely responsible for reabsorption of filtered glucose and gluconeogenesis (GNG). Insulin stimulates glucose transport and suppresses GNG in the proximal tubule, however, the signaling mechanisms and coordinated regulation of these processes remain poorly understood. The kinase complex mTORC2 is critical for regulation of growth, metabolism, solute transport, and electrolyte homeostasis in response to a wide array of inputs. Here we examined its role in the regulation of renal glucose reabsorption and GNG.

MethodsRictor, an essential component of mTORC2, was knocked out using the Pax8-LC1 system to generate inducible tubule specific Rictor knockout (TRKO) mice. These animals were subjected to fasting, refeeding, and variation in dietary K+. Metabolic parameters including glucose homeostasis and renal function were assessed in balance cages. Kidneys and livers were also harvested for molecular analysis of gluconeogenic enzymes, mTORC2-regulated targets, and plasma membrane glucose transporters.

ResultsOn a normal chow diet, TRKO mice had marked glycosuria despite indistinguishable blood glucose relative to WT controls. Kidney plasma membrane showed lower SGLT2 and SGLT1 in the fed state, supporting reduced renal glucose reabsorption. Additional metabolic testing provided evidence for renal insulin resistance with elevated fasting insulin, impaired pyruvate tolerance, elevated hemoglobin A1c, and increased renal gluconeogenic enzymes in the fasted and fed states. These effects were correlated with reduced downstream phosphorylation of Akt and the transcription factor FOXO4, identifying a novel role of FOXO4 in the kidney. Interestingly, high dietary K+ prevented glycosuria and excessive GNG in TRKO mice, despite persistent reduction in mTORC2 substrate phosphorylation.

ConclusionRenal tubule mTORC2 is critical for coordinated regulation of sodium-glucose cotransport by SGLT2 and SGLT1 as well as renal GNG. Dietary K+ promotes glucose reabsorption and suppresses GNG independently of insulin signaling and mTORC2, potentially providing an alternative signaling mechanism in states of insulin resistance.

SIGNIFICANCE STATEMENTThe kidney contributes to regulation of blood glucose through reabsorption of filtered glucose and gluconeogenesis. This study shows that mTORC2 and dietary potassium coordinate the regulation of sodium-glucose cotransport and glucose production in the kidney via independent mechanisms. New insights into the regulation of these processes in the kidney offer promising implications for diabetes mellitus management and treatment.
]]></description>
<dc:creator>Demko, J.</dc:creator>
<dc:creator>Saha, B.</dc:creator>
<dc:creator>Takagi, E.</dc:creator>
<dc:creator>Manis, A.</dc:creator>
<dc:creator>Weber, R.</dc:creator>
<dc:creator>Pearce, D.</dc:creator>
<dc:date>2024-06-27</dc:date>
<dc:identifier>doi:10.1101/2024.06.22.600201</dc:identifier>
<dc:title><![CDATA[Coordinated Regulation of Renal Glucose Reabsorption and Gluconeogenesis by mTORC2 and Potassium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.25.600671v1?rss=1">
<title>
<![CDATA[
Scalable intracellular delivery via microfluidic vortex shedding enhances the function of chimeric antigen receptor T-cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.25.600671v1?rss=1</link>
<description><![CDATA[
Adoptive chimeric antigen receptor T-cell (CAR-T) therapy is transformative and approved for hematologic malignancies. It is also being developed for the treatment of solid tumors, autoimmune disorders, heart disease, and aging. Despite unprecedented clinical outcomes, CAR-T and other engineered cell therapies face a variety of manufacturing and safety challenges. Traditional methods, such as lentivirus transduction and electroporation, result in random integration or cause significant cellular damage, which can limit the safety and efficacy of engineered cell therapies. We present hydroporation as a gentle and effective alternative for intracellular delivery. Hydroporation resulted in 1.7- to 2-fold higher CAR-T yields compared to electroporation with superior cell viability and recovery. Hydroporated cells exhibited rapid proliferation, robust target cell lysis, and increased pro-inflammatory and regulatory cytokine secretion in addition to improved CAR-T yield by day 5 post-transfection. We demonstrate that scaled-up hydroporation can process 5 x 108 cells in less than 10 s, showcasing the platform as a viable solution for high-yield CAR-T manufacturing with the potential for improved therapeutic outcomes.
]]></description>
<dc:creator>Sytsma, B. J.</dc:creator>
<dc:creator>Allain, V.</dc:creator>
<dc:creator>Bourke, S.</dc:creator>
<dc:creator>Faizee, F.</dc:creator>
<dc:creator>Fathi, M.</dc:creator>
<dc:creator>Berdeaux, R.</dc:creator>
<dc:creator>Ferreira, L. M. R.</dc:creator>
<dc:creator>Brewer, J.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Pan, F. L.</dc:creator>
<dc:creator>Rothrock, A. G.</dc:creator>
<dc:creator>Nyberg, W. A.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Wilson, L. H.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Pawell, R. S.</dc:creator>
<dc:date>2024-06-27</dc:date>
<dc:identifier>doi:10.1101/2024.06.25.600671</dc:identifier>
<dc:title><![CDATA[Scalable intracellular delivery via microfluidic vortex shedding enhances the function of chimeric antigen receptor T-cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.26.600491v1?rss=1">
<title>
<![CDATA[
Structure of the human K2P13.1(THIK-1) channel reveals a novel hydrophilic pore restriction and lipid cofactor site 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.26.600491v1?rss=1</link>
<description><![CDATA[
The halothane-inhibited K2P leak potassium channel K2P13.1 (THIK-1)1-3 is found in diverse cells1,4 including neurons1,5 and microglia6-8 where it affects surveillance6, synaptic pruning7, phagocytosis7, and inflammasome-mediated interleukin-1{beta} release6,8,9. As with many K2Ps1,5,10-14 and other voltage-gated ion channel (VGIC) superfamily members3,15,16, polyunsaturated fatty acid (PUFA) lipids modulate K2P13.1 (THIK-1)1,5,14,17 via a poorly understood mechanism. Here, we present cryo-electronmicroscopy (cryo-EM) structures of human K2P13.1 (THIK-1) and mutants in lipid nanodiscs and detergent. These reveal that, unlike other K2Ps13,18-24, K2P13.1 (THIK-1) has a two-chamber aqueous inner cavity obstructed by a M4 transmembrane helix tyrosine (Tyr273, the flow restrictor). This hydrophilic barrier can be opened by an activatory mutation, S136P25, at natural break in the M2 transmembrane helix and by intrinsic channel dynamics. The structures also reveal a buried lipid in the P1/M4 intersubunit interface at a location, the PUFA site, that coincides with the TREK subfamily K2P modulator pocket for small molecule agonists18,26,27. This overlap, together with the effects of mutation on K2P13.1 (THIK-1) PUFA responses, indicates that the PUFA site lipids are K2P13.1 (THIK-1) cofactors. Comparison with the PUFA-responsive VGIC Kv7.1 (KCNQ1)28-31 reveals a shared role for the equivalent pore domain intersubunit interface in lipid modulation, providing a framework for dissecting the effects of PUFAs on the VGIC superfamily. Our findings reveal the unique architecture underlying K2P13.1 (THIK-1) function, highlight the importance of the P1/M4 interface in control of K2Ps by both natural and synthetic agents, and should aid development of THIK subfamily modulators for diseases such as neuroinflammation6,32 and autism6.
]]></description>
<dc:creator>Roy-Chowdhury, S.</dc:creator>
<dc:creator>Jang, S.</dc:creator>
<dc:creator>Abderemane-Ali, F.</dc:creator>
<dc:creator>Naughton, F.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>Minor, D. L.</dc:creator>
<dc:date>2024-06-27</dc:date>
<dc:identifier>doi:10.1101/2024.06.26.600491</dc:identifier>
<dc:title><![CDATA[Structure of the human K2P13.1(THIK-1) channel reveals a novel hydrophilic pore restriction and lipid cofactor site]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.27.600921v1?rss=1">
<title>
<![CDATA[
Long-term in vitro exposure of Treponema pallidum to sub-bactericidal doxycycline did not induce resistance: Implications for doxy-PEP and syphilis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.27.600921v1?rss=1</link>
<description><![CDATA[
Doxycycline post-exposure prophylaxis (doxy-PEP) could significantly reduce syphilis incidence. However, the increase in intermittent doxycycline usage might select resistant Treponema pallidum (T. pallidum) strains. To assess whether resistance to doxycycline could be induced in this pathogen, we exposed the SS14 strain in vitro both intermittently and continuously to a sub-bactericidal doxycycline concentration that still exerts antibiotic pressure. During and after each exposure experiment, we assessed the doxycycline minimal inhibitory concentration in test and control treponemes and performed whole genome sequencing, concluding that no resistance developed. This work suggests that doxycycline-resistant T. pallidum is not an immediate threat for doxy-PEP implementation.
]]></description>
<dc:creator>Tantalo, L. C.</dc:creator>
<dc:creator>Luetkemeyer, A.</dc:creator>
<dc:creator>Lieberman, N. A. P.</dc:creator>
<dc:creator>Nunley, E.</dc:creator>
<dc:creator>Avendano, C.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>Celum, C.</dc:creator>
<dc:creator>Giacani, L.</dc:creator>
<dc:date>2024-06-27</dc:date>
<dc:identifier>doi:10.1101/2024.06.27.600921</dc:identifier>
<dc:title><![CDATA[Long-term in vitro exposure of Treponema pallidum to sub-bactericidal doxycycline did not induce resistance: Implications for doxy-PEP and syphilis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.27.600910v1?rss=1">
<title>
<![CDATA[
TOMM40 regulates hepatocellular and plasma lipid metabolism via an LXR-dependent pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.27.600910v1?rss=1</link>
<description><![CDATA[
The gene encoding TOMM40 (Transporter of Outer Mitochondrial Membrane 40) is adjacent to that encoding APOE, which has a central role in lipid and lipoprotein metabolism. Human genetic variants near APOE and TOMM40 are strongly associated with plasma lipid levels, but a specific role for TOMM40 in lipid metabolism has not been established. Investigating this, we show that suppression of TOMM40 in human hepatoma cells upregulates expression of APOE and LDLR in part via activation of LXRB (NR1H2) by oxysterols, with consequent increased uptake of VLDL and LDL. This is in part due to disruption of mitochondria-endoplasmic reticulum contact sites, with resulting accrual of reactive oxygen species and non-enzymatically derived oxysterols. With TOMM40 knockdown, cellular triglyceride and lipid droplet content are increased, effects attributable in part to receptor-mediated VLDL uptake, since lipid staining is significantly reduced by concomitant suppression of either LDLR or APOE. In contrast, cellular cholesterol content is reduced due to LXRB-mediated upregulation of the ABCA1 transporter as well as increased production and secretion of oxysterol-derived cholic acid. Consistent with the findings in hepatoma cells, in vivo knockdown of TOMM40 in mice results in significant reductions of plasma triglyceride and cholesterol concentrations, reduced hepatic cholesterol and increased triglyceride content, and accumulation of lipid droplets leading to development of steatosis. These findings demonstrate a role for TOMM40 in regulating hepatic lipid and plasma lipoprotein levels and identify mechanisms linking mitochondrial function with lipid metabolism.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=132 SRC="FIGDIR/small/600910v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Yang, N. V.</dc:creator>
<dc:creator>Chao, J. Y.</dc:creator>
<dc:creator>Garton, K. A.</dc:creator>
<dc:creator>Tran, T.</dc:creator>
<dc:creator>King, S.</dc:creator>
<dc:creator>Orr, J.</dc:creator>
<dc:creator>Oei, J. H.</dc:creator>
<dc:creator>Kang, M.</dc:creator>
<dc:creator>Zalpuri, R.</dc:creator>
<dc:creator>Jorgens, D. M.</dc:creator>
<dc:creator>Konchadi, P.</dc:creator>
<dc:creator>Chorba, J. S.</dc:creator>
<dc:creator>Theusch, E.</dc:creator>
<dc:creator>Krauss, R. M.</dc:creator>
<dc:date>2024-06-29</dc:date>
<dc:identifier>doi:10.1101/2024.06.27.600910</dc:identifier>
<dc:title><![CDATA[TOMM40 regulates hepatocellular and plasma lipid metabolism via an LXR-dependent pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.27.600934v1?rss=1">
<title>
<![CDATA[
Improved detection of microbiome-disease associations via population structure-aware generalized linear mixed effects models (microSLAM) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.27.600934v1?rss=1</link>
<description><![CDATA[
Microbiome association studies typically link host disease or other traits to summary statistics measured in metagenomics data, such as diversity or taxonomic composition. But identifying disease-associated species based on their relative abundance does not provide insight into why these microbes act as disease markers, and it overlooks cases where disease risk is related to specific strains with unique biological functions. To bridge this knowledge gap, we developed microSLAM, a mixed-effects model and an R package that performs association tests that connect host traits to the presence/absence of genes within each microbiome species, while accounting for strain genetic relatedness across hosts. Traits can be quantitative or binary (such as case/control). MicroSLAM is fit in three steps for each species. The first step estimates population structure across hosts. Step two calculates the association between population structure and the trait, enabling detection of species for which a subset of related strains confer risk. To identify specific genes whose presence/absence across diverse strains is associated with the trait, step three models the trait as a function of gene occurrence plus random effects estimated from step two. Applying microSLAM to 710 gut metagenomes from inflammatory bowel disease (IBD) samples, we discovered 49 species whose population structure correlates with IBD. In addition, after controlling for population structure, we found 57 microbial genes that are significantly more common in healthy individuals and 26 that are more common in IBD patients, including a seven-gene operon in Faecalibacterium prausnitzii that is involved in utilization of fructoselysine from the gut environment. Overall, microSLAM detected IBD associations for 45 species that were not detected using relative abundance tests, and it identified specific strains and genes underlying IBD associations for 13 other species. These findings highlight the importance of accounting for within-species genetic variation in microbiome studies.

Author SummaryThe species composition of the human gut microbiome differs significantly between individuals and is associated with various diseases. Many studies have sought to understand this relationship by examining the relative amount of each bacterial species within metagenomic sequencing data from sick and healthy individuals. However, this approach makes it challenging to pinpoint which genes and pathways of a disease-associated species might actually contribute to disease risk, and it misses species where only certain strains are disease associated. To overcome these challenges, we developed an R package, called microSLAM, that uses mixed-effects modeling to associate within-species genetic variation to host traits. microSLAM performs two types of tests for each species: one for identifying strain-disease associations and another for identifying gene-disease associations. The gene tests account for the genetic relatedness of strains across hosts, making them particularly useful for detecting mobile genes. We applied microSLAM to hundreds of gut metagenomes from inflammatory bowel disease studies, identifying dozens of novel associations that were missed using relative abundance tests. MicroSLAM is a general modeling approach that can be applied to human traits beyond disease case/control studies and to microbiomes from other environments.
]]></description>
<dc:creator>Goldman, M.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2024-06-30</dc:date>
<dc:identifier>doi:10.1101/2024.06.27.600934</dc:identifier>
<dc:title><![CDATA[Improved detection of microbiome-disease associations via population structure-aware generalized linear mixed effects models (microSLAM)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.27.600908v1?rss=1">
<title>
<![CDATA[
Peripheral glia and neurons jointly regulate activity-induced synaptic remodeling at the Drosophila neuromuscular junction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.27.600908v1?rss=1</link>
<description><![CDATA[
In the nervous system, reliable communication depends on the ability of neurons to adaptively remodel their synaptic structure and function in response to changes in neuronal activity. While neurons are the main drivers of synaptic plasticity, glial cells are increasingly recognized for their roles as active modulators. However, the underlying molecular mechanisms remain unclear. Here, using Drosophila neuromuscular junction as a model system for a tripartite synapse, we show that peripheral glial cells collaborate with neurons at the NMJ to regulate activity-induced synaptic remodeling, in part through a protein called shriveled (Shv). Shv is an activator of integrin signaling previously shown to be released by neurons during intense stimulation at the fly NMJ to regulate activity-induced synaptic remodeling. We demonstrate that Shv is also present in peripheral glia, and glial Shv is both necessary and sufficient for synaptic remodeling. However, unlike neuronal Shv, glial Shv does not activate integrin signaling at the NMJ. Instead, it regulates synaptic plasticity in two ways: 1) maintaining the extracellular balance of neuronal Shv proteins to regulate integrin signaling, and 2) controlling ambient extracellular glutamate concentration to regulate postsynaptic glutamate receptor abundance. Loss of glial cells showed the same phenotype as loss of Shv in glia. Together, these results reveal that neurons and glial cells homeostatically regulate extracellular Shv protein levels to control activity-induced synaptic remodeling. Additionally, peripheral glia maintains postsynaptic glutamate receptor abundance and contribute to activity-induced synaptic remodeling by regulating ambient glutamate concentration at the fly NMJ.
]]></description>
<dc:creator>Chang, Y.-C.</dc:creator>
<dc:creator>Peng, Y.-J.</dc:creator>
<dc:creator>Lee, J. Y.</dc:creator>
<dc:creator>Chang, K. T.</dc:creator>
<dc:date>2024-07-02</dc:date>
<dc:identifier>doi:10.1101/2024.06.27.600908</dc:identifier>
<dc:title><![CDATA[Peripheral glia and neurons jointly regulate activity-induced synaptic remodeling at the Drosophila neuromuscular junction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.27.601077v1?rss=1">
<title>
<![CDATA[
Uncovering differential tolerance to deletions versus substitutions with a protein language model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.27.601077v1?rss=1</link>
<description><![CDATA[
Deep mutational scanning (DMS) experiments have been successfully leveraged to understand genotype to phenotype mapping, with broad implications for protein engineering, human genetics, drug development, and beyond. To date, however, the overwhelming majority of DMS have focused on amino acid substitutions, excluding other classes of variation such as deletions or insertions. As a consequence, it remains unclear how indels differentially shape the fitness landscape relative to substitutions. In order to further our understanding of the relationship between substitutions and deletions, we leveraged a protein language model to analyze every single amino acid deletion in the human proteome. We discovered hundreds of thousands of sites that display opposing behavior for deletions versus substitutions, i.e. sites that can tolerate being substituted but not deleted, and vice versa. We identified secondary structural elements and sequence context to be important mediators of differential tolerability at these sites. Our results underscore the value of deletion-substitution comparisons at the genome-wide scale, provide novel insights into how substitutions could systematically differ from deletions, and showcase the power of protein language models to generate biological hypotheses in-silico. All deletion-substitution comparisons can be explored and downloaded at https://huggingface.co/spaces/ntranoslab/diff-tol.
]]></description>
<dc:creator>Goldman, G.</dc:creator>
<dc:creator>Chati, P.</dc:creator>
<dc:creator>Ntranos, V.</dc:creator>
<dc:date>2024-07-02</dc:date>
<dc:identifier>doi:10.1101/2024.06.27.601077</dc:identifier>
<dc:title><![CDATA[Uncovering differential tolerance to deletions versus substitutions with a protein language model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.28.600695v1?rss=1">
<title>
<![CDATA[
Neural Stem Cell Relay from B1 to B2 cells in the adult mouse Ventricular-Subventricular Zone 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.28.600695v1?rss=1</link>
<description><![CDATA[
Neurogenesis and gliogenesis continue in the Ventricular-Subventricular Zone (V-SVZ) of the adult rodent brain. B1 cells are astroglial cells derived from radial glia that function as primary progenitors or neural stem cells (NSCs) in the V-SVZ. B1 cells, which have a small apical contact with the ventricle, decline in numbers during early postnatal life, yet neurogenesis continues into adulthood. Here we found that a second population of V-SVZ astroglial cells (B2 cells), that do not contact the ventricle, function as NSCs in the adult brain. B2 cell numbers increase postnatally, remain constant in 12-month-old mice and decrease by 18 months. Transcriptomic analysis of ventricular-contacting and non-contacting B cells revealed key molecular differences to distinguish B1 from B2 cells. Transplantation and lineage tracing of B2 cells demonstrate their function as primary progenitors for adult neurogenesis. This study reveals how NSC function is relayed from B1 to B2 progenitors to maintain adult neurogenesis.
]]></description>
<dc:creator>Cebrian-Silla, A.</dc:creator>
<dc:creator>Assis Nascimento, M.</dc:creator>
<dc:creator>Mancia Leon, W.</dc:creator>
<dc:creator>Gonzalez-Granero, S.</dc:creator>
<dc:creator>Romero-Rodriguez, R.</dc:creator>
<dc:creator>Obernier, K.</dc:creator>
<dc:creator>Steffen, D. M.</dc:creator>
<dc:creator>Lim, D. A.</dc:creator>
<dc:creator>Garcia-Verdugo, J. M.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:date>2024-07-02</dc:date>
<dc:identifier>doi:10.1101/2024.06.28.600695</dc:identifier>
<dc:title><![CDATA[Neural Stem Cell Relay from B1 to B2 cells in the adult mouse Ventricular-Subventricular Zone]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.01.601579v1?rss=1">
<title>
<![CDATA[
HIV-1 control in vivo is related to the number but not the fraction of infected cells with viral unspliced RNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.01.601579v1?rss=1</link>
<description><![CDATA[
In the absence of antiretroviral therapy (ART), a subset of individuals, termed HIV controllers, have levels of plasma viremia that are orders of magnitude lower than non-controllers who are at higher risk for HIV disease progression. In addition to having fewer infected cells resulting in fewer cells with HIV RNA, it is possible that lower levels of plasma viremia in controllers is due to a lower fraction of the infected cells having HIV-1 unspliced RNA (HIV usRNA) compared with non-controllers. To directly test this possibility, we used sensitive and quantitative single cell sequencing methods to compare the fraction of infected cells that contain one or more copies of HIV usRNA in peripheral blood mononuclear cells (PBMC) obtained from controllers and non-controllers. The fraction of infected cells containing HIV usRNA did not differ between the two groups. Rather, the levels of viremia were strongly associated with the total number of infected cells that had HIV usRNA, as reported by others, with controllers having 34-fold fewer infected cells per million PBMC. These results reveal for the first time that viremic control is not associated with a lower fraction of proviruses expressing HIV usRNA, unlike what is reported for elite controllers, but is only related to having fewer infected cells overall, maybe reflecting greater immune clearance of infected cells. Our findings show that proviral silencing is not a key mechanism for viremic control and will help to refine strategies towards achieving HIV remission without ART.
]]></description>
<dc:creator>Capoferri, A. A.</dc:creator>
<dc:creator>Wiegand, A.</dc:creator>
<dc:creator>Hong, F.</dc:creator>
<dc:creator>Jacobs, J. L.</dc:creator>
<dc:creator>Spindler, J.</dc:creator>
<dc:creator>Musick, A.</dc:creator>
<dc:creator>Bale, M. J.</dc:creator>
<dc:creator>Shao, W.</dc:creator>
<dc:creator>Sobolewski, M. D.</dc:creator>
<dc:creator>Cillo, A. R.</dc:creator>
<dc:creator>Luke, B. T.</dc:creator>
<dc:creator>Fennessey, C. M.</dc:creator>
<dc:creator>Gorelick, R. J.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Halvas, E. K.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Coffin, J. M.</dc:creator>
<dc:creator>Mellors, J. W.</dc:creator>
<dc:creator>Kearney, M. F.</dc:creator>
<dc:date>2024-07-02</dc:date>
<dc:identifier>doi:10.1101/2024.07.01.601579</dc:identifier>
<dc:title><![CDATA[HIV-1 control in vivo is related to the number but not the fraction of infected cells with viral unspliced RNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.01.601334v1?rss=1">
<title>
<![CDATA[
Multiplatform molecular profiling and functional genomic screens identify prognostic signatures and mechanisms underlying MEK inhibitor response in somatic NF1 mutant glioblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.01.601334v1?rss=1</link>
<description><![CDATA[
NF1 is recurrently mutated in glioblastoma yet the molecular landscape and efficacy of targeted therapies remain unclear. Here, we combine bulk and single cell genomics of human somatic NF1 mutant, IDH-wildtype glioblastomas with functional genomic analysis of cell lines and mouse intracranial tumor models to identify molecular subgroups within NF1 mutant glioblastomas and mechanisms underlying MEK inhibitor response. Targeted DNA sequencing showed homozygous deletion of the cell cycle regulator CDKN2A/B is a poor prognostic marker in somatic NF1 mutant, but not NF1 wildtype, tumors. DNA methylation array profiling revealed three epigenetic groups highlighted by distinct clinical features, co-mutation patterns, and reference methylation classifier identities. Genome-wide CRISPRi screens in glioblastoma cells revealed cell cycle regulators are conserved mediators of cell growth while response to the MEK inhibitor selumetinib converges on Ras/RAF/MEK pathway activation. Repression of the RAF regulator SHOC2 sensitizes glioblastomas to selumetinib in vitro and in vivo in mouse intracranial glioblastoma models. Single cell RNA-sequencing of mouse intracranial glioblastomas treated with the MEK inhibitor selumetinib reveals distinct responses between mesenchymal-like (MES-like) and non MES-like subpopulations suggesting non-MES like cells are intrinsically resistant to MEK inhibition. Finally, single nuclear RNA-sequencing (snRNA-seq) of human NF1 mutant, CDKN2A/B deleted glioblastomas reveals MES-like tumor cells are associated with selumetinib sensitivity signatures while non MES-like cells exhibit increased cell cycle progression and lack selumetinib sensitivity, further supporting the notion that MEK inhibition specifically targets MES-like tumor cell subpopulations. Taken together, our data underscores the heterogeneity between and within somatic NF1 mutant glioblastomas and delineates mechanisms of MEK inhibitor response across distinct tumor subpopulations, guiding the development of future therapeutic strategies that may synergize with MEK inhibition for NF1 mutant tumors.

The tumor suppressor NF1 is mutated in 15% of glioblastomas,1-3 the most common malignant brain tumor with poor outcomes and few effective treatments.4 NF1 is a GTPase activating protein (GAP) that negatively regulates Ras, and thus, NF1 loss leads to induction of Ras/RAF/MEK/ERK signaling, driving tumorigenesis and comprising a targetable molecular cascade.5,6 Genomic analysis of glioblastoma demonstrates NF1 mutation is associated with a mesenchymal-like (MES-like) transcriptomic tumor cell subpopulation and altered tumor microenvironment.7,8 More broadly, DNA methylation analysis reveals multiple epigenetic subgroups with overlapping relationships to transcriptomic subtype and DNA alterations, underscoring the complex relationship between genetic drivers and molecular signatures.9 While the updated 2021 Central Nervous System WHO tumor classification incorporates an ever increasing amount of molecular criteria for diffuse astrocytic tumors,10 the existence and clinical significance of molecular subgroups within somatic NF1 mutant, IDH-wildtype glioblastomas based on genetic co-mutations, epigenetic profile, or transcriptomic signatures remain unclear.

Preclinical data support the utility of MEK inhibition in NF1 mutant gliomas,11,12 and the MEK inhibitor selumetinib is FDA approved for tumors arising in patients with syndromic neurofibromatosis type 1 (NF-1) harboring a germline NF1 mutation.13,14 In NF-1 associated gliomas, MEK inhibition demonstrates efficacy in a limited case series,15 and combined BRAF/MEK inhibition shows efficacy in BRAF p.V600E mutant gliomas,16 further supporting the translational potential of Ras/Raf/MEK/ERK blockade within genetically defined glioma subtypes. Nevertheless, treatment resistance to molecular monotherapy remains a challenge,17-20 and the mechanisms underlying MEK inhibitor resistance in NF1 mutant glioma are unknown.

Here, we integrate targeted DNA sequencing, DNA methylation profiling, and single nuclear RNA-sequencing (snRNA-seq) of human patient somatic NF1 mutant, IDH-wildtype glioblastomas with single cell RNA-sequencing (scRNA-seq), genome-wide clustered regularly interspaced short palindromic repeats interference (CRISPRi) screens, and pharmacologic studies in cell lines and mouse intracranial glioblastoma models to define molecular subgroups and functional mediators of MEK inhibitor response. Targeted DNA sequencing of NF1 mutant, IDH-wildtype glioblastomas (n=186 tumors) revealed CDKN2A/B deletion was associated with poor outcomes in NF1 mutant, but not NF1 wildtype, glioblastomas. DNA methylation profiling (n=129 tumors) demonstrated three epigenetic subgroups with distinct clinical features, co- mutation patterns across cell cycle genes, and reference methylation classifier identities.

Genome-wide CRISPRi screens in mouse SB28 and human GBM43 glioblastoma cells identified a conserved cell cycle gene network mediating cell growth, consistent with the clinical importance of additional hits affecting the cell cycle in human somatic NF1 mutant glioblastomas. Moreover, genome-wide mediators of selumetinib response converged upon two Ras pathway effectors mediating selumetinib sensitivity: BRAF and SHOC2. SHOC2 repression in glioblastoma cells significantly improved selumetinib response both in vitro and in intracranial allografts in vivo. Single cell RNA-sequencing (scRNA-seq) of mouse intracranial glioblastomas treated with the MEK inhibitor selumetinib revealed MES-like tumor cells correlated with CDKN2A retention and the CRISPRi screen selumetinib sensitivity signature, with selumetinib resistant cells displaying Ras pathway induction. In contrast, non-MES like tumor cells were CDKN2A deficient and lacked expression of the CRISPRi screen selumetinib sensitivity signature, with selumetinib resistant cells inducing a glial de-differentiation program. Finally, snRNA-seq of NF1 mutant, CDKN2A/B deleted, IDH-wildtype glioblastomas (n=9) showed non MES-like tumor cells exhibit increased cell cycle progression and were not associated with the CRISPRi screen selumetinib sensitivity signature. MES-like tumor cells within newly diagnosed, but not recurrent, tumors retained expression of the CRISPRi screen selumetinib sensitivity signature, suggesting resistance can arise both between and within specific transcriptomic glioblastoma cell tumor cell subpopulations. Taken together, our data identifies clinically important subgroups of NF1 mutant, IDH-wildtype glioblastomas and supports a model in which heterogeneity between tumors and within tumor cell subpopulations underlies MEK inhibitor response, supporting the need for additional synergistic therapeutic approaches beyond maximal Ras pathway blockade for NF1 mutant glioblastomas.
]]></description>
<dc:creator>Pan, S.</dc:creator>
<dc:creator>Mirchia, K.</dc:creator>
<dc:creator>Payne, E.</dc:creator>
<dc:creator>Liu, S. J.</dc:creator>
<dc:creator>Al-Adli, N.</dc:creator>
<dc:creator>Peeran, Z.</dc:creator>
<dc:creator>Shukla, P.</dc:creator>
<dc:creator>Young, J. S.</dc:creator>
<dc:creator>Gupta, R.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Pak, J.</dc:creator>
<dc:creator>Seo, K.</dc:creator>
<dc:creator>Ozawa, T.</dc:creator>
<dc:creator>Na, B.</dc:creator>
<dc:creator>Reddy, A. T.</dc:creator>
<dc:creator>Braunstein, S. E.</dc:creator>
<dc:creator>Phillips, J. J.</dc:creator>
<dc:creator>Chang, S.</dc:creator>
<dc:creator>Solomon, D. A.</dc:creator>
<dc:creator>Perry, A.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:creator>Berger, M. S.</dc:creator>
<dc:creator>Abate, A. R.</dc:creator>
<dc:creator>Vasudevan, H. N.</dc:creator>
<dc:date>2024-07-03</dc:date>
<dc:identifier>doi:10.1101/2024.07.01.601334</dc:identifier>
<dc:title><![CDATA[Multiplatform molecular profiling and functional genomic screens identify prognostic signatures and mechanisms underlying MEK inhibitor response in somatic NF1 mutant glioblastoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.01.601474v1?rss=1">
<title>
<![CDATA[
The Neurolipid Atlas: a lipidomics resource for neurodegenerative diseases uncovers cholesterol as a regulator of astrocyte reactivity impaired by ApoE4 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.01.601474v1?rss=1</link>
<description><![CDATA[
Lipid changes in the brain have been implicated in many neurodegenerative diseases including Alzheimers Disease (AD), Parkinsons disease and Amyotrophic Lateral Sclerosis. To facilitate comparative lipidomic research across brain-diseases we established a data commons named the Neurolipid Atlas, that we have pre-populated with novel human, mouse and isogenic induced pluripotent stem cell (iPSC)-derived lipidomics data for different brain diseases. We show that iPSC-derived neurons, microglia and astrocytes display distinct lipid profiles that recapitulate in vivo lipotypes. Leveraging multiple datasets, we show that the AD risk gene ApoE4 drives cholesterol ester (CE) accumulation in human astrocytes recapitulating CE accumulation measured in the human AD brain. Multi-omic interrogation of iPSC-derived astrocytes revealed that cholesterol plays a major role in astrocyte interferon-dependent pathways such as the immunoproteasome and major histocompatibility complex (MHC) class I antigen presentation. We show that through enhanced cholesterol esterification ApoE4 suppresses immune activation of astrocytes. Our novel data commons, available at neurolipidatlas.com, provides a user-friendly tool and knowledge base for a better understanding of lipid dyshomeostasis in neurodegenerative diseases.
]]></description>
<dc:creator>Feringa, F. M.</dc:creator>
<dc:creator>Koppes-den Hertog, S. J.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Derks, R. J. E.</dc:creator>
<dc:creator>Kruijff, I.</dc:creator>
<dc:creator>Erlebach, L.</dc:creator>
<dc:creator>Heijneman, J.</dc:creator>
<dc:creator>Miramontes, R.</dc:creator>
<dc:creator>Pömpner, N.</dc:creator>
<dc:creator>Blomberg, N.</dc:creator>
<dc:creator>Olivier-Jimenez, D.</dc:creator>
<dc:creator>Johanson, L. E.</dc:creator>
<dc:creator>Cammack, A. J.</dc:creator>
<dc:creator>Giblin, A.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Rose, I. V. L.</dc:creator>
<dc:creator>Yuan, H.</dc:creator>
<dc:creator>Ward, M. E.</dc:creator>
<dc:creator>Isaacs, A.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Kronenberg-Versteeg, D.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Thompson, L. M.</dc:creator>
<dc:creator>Ori, A.</dc:creator>
<dc:creator>Mohammed, Y.</dc:creator>
<dc:creator>Giera, M.</dc:creator>
<dc:creator>van der Kant, R. H. N.</dc:creator>
<dc:date>2024-07-03</dc:date>
<dc:identifier>doi:10.1101/2024.07.01.601474</dc:identifier>
<dc:title><![CDATA[The Neurolipid Atlas: a lipidomics resource for neurodegenerative diseases uncovers cholesterol as a regulator of astrocyte reactivity impaired by ApoE4]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.01.601365v1?rss=1">
<title>
<![CDATA[
Structure of a novel α-synuclein filament fold from multiple system atrophy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.01.601365v1?rss=1</link>
<description><![CDATA[
Multiple system atrophy (MSA) is a synucleinopathy, a group of related diseases characterized by the accumulation of -synuclein aggregates in the brain. In MSA, these aggregates form glial cytoplasmic inclusions, which contain abundant cross-{beta} amyloid filaments. Structures of -synuclein filaments isolated from MSA patient tissue were determined by cryo-electron microscopy (cryo-EM), revealing three discrete folds that are distinct from -synuclein filaments associated with other synucleinopathies. Here, we use cryo-EM classification methods to characterize filaments from one individual with MSA and identify a novel, low-populated MSA filament fold (designated Type I2) in addition to a predominant class comprising MSA Type II2. The 3.3-[A] resolution structure of the Type I2 filament reveals a fold consisting of two asymmetric protofilaments. One is identical to a previously solved Type I protofilament, while the second adopts a novel fold that is chimeric between two previously reported Type I and II protofilaments. These results further define disease-specific folds of -synuclein filaments that develop in MSA and have implications for the design of therapeutic and diagnostic molecules that target disease.
]]></description>
<dc:creator>Yan, N. L.</dc:creator>
<dc:creator>Candido, F.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Melo, A. A.</dc:creator>
<dc:creator>Prusiner, S. B.</dc:creator>
<dc:creator>Mordes, D. A.</dc:creator>
<dc:creator>Southworth, D. R.</dc:creator>
<dc:creator>Paras, N. A.</dc:creator>
<dc:creator>Merz, G. E.</dc:creator>
<dc:date>2024-07-04</dc:date>
<dc:identifier>doi:10.1101/2024.07.01.601365</dc:identifier>
<dc:title><![CDATA[Structure of a novel α-synuclein filament fold from multiple system atrophy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.02.601671v1?rss=1">
<title>
<![CDATA[
iTome Volumetric Serial Sectioning Apparatus for TEM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.02.601671v1?rss=1</link>
<description><![CDATA[
An automated ultra-microtome capable of sectioning thousands of ultrathin sections onto standard TEM slot grids was developed and used to section: a complete Drosophila melanogaster first-instar larva, three sections per grid, into 4,866 34-nm-thick sections with a cutting and pickup success rate of 99.74%; 30 microns of mouse cortex measuring roughly 400 um x 2000 um at 40 nm per slice; and a full adult Drosophila brain and ventral nerve column into 9,300 sections with a pickup success rate of 99.95%. The apparatus uses optical interferometers to monitor a reference distance between the cutting knife and multiple sample blocks. Cut sections are picked up from the knife-boat water surface while they are still anchored to the cutting knife. Blocks without embedded tissue are used to displace tissue-containing sections away from the knife edge so that the tissue regions end up in the grid slot instead of on the grid rim.
]]></description>
<dc:creator>Peale, D. R.</dc:creator>
<dc:creator>Hess, H.</dc:creator>
<dc:creator>Lee, P. R.</dc:creator>
<dc:creator>Cardona, A.</dc:creator>
<dc:creator>Bock, D.</dc:creator>
<dc:creator>Schneider-Mizell, C. M.</dc:creator>
<dc:creator>Fetter, R. D.</dc:creator>
<dc:creator>Lee, W.-P.</dc:creator>
<dc:creator>Robinson, C. G.</dc:creator>
<dc:creator>Iyer, N.</dc:creator>
<dc:creator>Managan, C.</dc:creator>
<dc:date>2024-07-04</dc:date>
<dc:identifier>doi:10.1101/2024.07.02.601671</dc:identifier>
<dc:title><![CDATA[iTome Volumetric Serial Sectioning Apparatus for TEM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.02.601752v1?rss=1">
<title>
<![CDATA[
Microglia mediate the early-life programming of adult glucose control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.02.601752v1?rss=1</link>
<description><![CDATA[
Mammalian glucose homeostasis is, in part, nutritionally programmed during early neonatal life, a critical window for the formation of synapses between hypothalamic glucoregulatory centers. Although microglia are known to prune synapses throughout the brain, their specific role in refining hypothalamic glucoregulatory circuits remains unknown. Here, we show that microglia in the mediobasal hypothalamus (MBH) of mice actively engage in synaptic pruning during early life. Microglial phagocytic activity is induced following birth, regresses upon weaning from maternal milk, and is exacerbated by feeding dams a high-fat diet while lactating. In particular, we show that microglia refine perineuronal nets (PNNs) within the neonatal MBH. Indeed, transiently depleting microglia before weaning (P6-16), but not afterward (P21-31), remarkably increased PNN abundance in the MBH. Furthermore, mice lacking microglia only from P6-16 had glucose intolerance due to impaired glucose-responsive pancreatic insulin secretion in adulthood, a phenotype not seen if microglial depletion occurred after weaning. Viral retrograde tracing revealed that this impairment is linked to a reduction in the number of neurons in specific hypothalamic glucoregulatory centers that synaptically connect to the pancreatic {beta}-cell compartment. These findings show that microglia facilitate synaptic plasticity in the MBH during early life through a process that includes PNN refinement, to establish hypothalamic circuits that regulate adult glucose homeostasis.
]]></description>
<dc:creator>Valdearcos, M.</dc:creator>
<dc:creator>McGrath, E. R.</dc:creator>
<dc:creator>Brown Mayfield, S.</dc:creator>
<dc:creator>Folick, A.</dc:creator>
<dc:creator>Cheang, R. T.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Bachor, T. P.</dc:creator>
<dc:creator>Lippert, R. N.</dc:creator>
<dc:creator>Xu, A. W.</dc:creator>
<dc:creator>Koliwad, S. K.</dc:creator>
<dc:date>2024-07-04</dc:date>
<dc:identifier>doi:10.1101/2024.07.02.601752</dc:identifier>
<dc:title><![CDATA[Microglia mediate the early-life programming of adult glucose control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.08.600855v1?rss=1">
<title>
<![CDATA[
The WAVE complex forms linear arrays at negative membrane curvature to instruct lamellipodia formation. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.08.600855v1?rss=1</link>
<description><![CDATA[
Cells generate a wide range of actin-based membrane protrusions for various cell behaviors. These protrusions are organized by different actin nucleation promoting factors. For example, N-WASP controls finger-like filopodia, whereas the WAVE complex controls sheet-like lamellipodia. These different membrane morphologies likely reflect different patterns of nucleator self-organization. N-WASP phase separation has been successfully studied through biochemical reconstitutions, but how the WAVE complex self-organizes to instruct lamellipodia is unknown. Because WAVE complex self-organization has proven refractory to cell-free studies, we leverage in vivo biochemical approaches to investigate WAVE complex organization within its native cellular context. With single molecule tracking and molecular counting, we show that the WAVE complex forms highly regular multilayered linear arrays at the plasma membrane that are reminiscent of a microtubule-like organization. Similar to the organization of microtubule protofilaments in a curved array, membrane curvature is both necessary and sufficient for formation of these WAVE complex linear arrays, though actin polymerization is not. This dependency on negative membrane curvature could explain both the templating of lamellipodia and their emergent behaviors, including barrier avoidance. Our data uncover the key biophysical properties of mesoscale WAVE complex patterning and highlight an integral relationship between NPF self-organization and cell morphogenesis.
]]></description>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Marchando, P.</dc:creator>
<dc:creator>Meyer, K.</dc:creator>
<dc:creator>Tang, Z.</dc:creator>
<dc:creator>Woolfson, D. N.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:date>2024-07-08</dc:date>
<dc:identifier>doi:10.1101/2024.07.08.600855</dc:identifier>
<dc:title><![CDATA[The WAVE complex forms linear arrays at negative membrane curvature to instruct lamellipodia formation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.05.602265v1?rss=1">
<title>
<![CDATA[
Current genomic deep learning models display decreased performance in cell type specific accessible regions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.05.602265v1?rss=1</link>
<description><![CDATA[
AbstractO_ST_ABSBackgroundC_ST_ABSA number of deep learning models have been developed to predict epigenetic features such as chromatin accessibility from DNA sequence. Model evaluations commonly report performance genome-wide; however, cis regulatory elements (CREs), which play critical roles in gene regulation, make up only a small fraction of the genome. Furthermore, cell type specific CREs contain a large proportion of complex disease heritability.

ResultsWe evaluate genomic deep learning models in chromatin accessibility regions with varying degrees of cell type specificity. We assess two modeling directions in the field: general purpose models trained across thousands of outputs (cell types and epigenetic marks), and models tailored to specific tissues and tasks. We find that the accuracy of genomic deep learning models, including two state-of-the-art general purpose models - Enformer and Sei - varies across the genome and is reduced in cell type specific accessible regions. Using accessibility models trained on cell types from specific tissues, we find that increasing model capacity to learn cell type specific regulatory syntax - through single-task learning or high capacity multi-task models - can improve performance in cell type specific accessible regions. We also observe that improving reference sequence predictions does not consistently improve variant effect predictions, indicating that novel strategies are needed to improve performance on variants.

ConclusionsOur results provide a new perspective on the performance of genomic deep learning models, showing that performance varies across the genome and is particularly reduced in cell type specific accessible regions. We also identify strategies to maximize performance in cell type specific accessible regions.
]]></description>
<dc:creator>Kathail, P.</dc:creator>
<dc:creator>Shuai, R. W.</dc:creator>
<dc:creator>Chung, R.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Loeb, G.</dc:creator>
<dc:creator>Ioannidis, N. M.</dc:creator>
<dc:date>2024-07-10</dc:date>
<dc:identifier>doi:10.1101/2024.07.05.602265</dc:identifier>
<dc:title><![CDATA[Current genomic deep learning models display decreased performance in cell type specific accessible regions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.08.602536v1?rss=1">
<title>
<![CDATA[
The impact of Library Size and Scale of Testing on Virtual Screening 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.08.602536v1?rss=1</link>
<description><![CDATA[
Virtual libraries for ligand discovery have recently increased 10,000-fold, and this is thought to have improved hit rates and potencies from library docking. This idea has not, however, been experimentally tested in direct comparisons of larger-vs-smaller libraries. Meanwhile, though libraries have exploded, the scale of experimental testing has little changed, with often only dozens of high-ranked molecules investigated, making interpretation of hit rates and affinities uncertain. Accordingly, we docked a 1.7 billion molecule virtual library against the model enzyme AmpC {beta}-lactamase, testing 1,521 new molecules and comparing the results to the same screen with a library of 99 million molecules, where only 44 molecules were tested. Encouragingly, the larger screen outperformed the smaller one: hit rates improved by two-fold, more new scaffolds were discovered, and potency improved. Overall, 50-fold more inhibitors were found, supporting the idea that there are many more compounds to be discovered than are being tested. With so many compounds evaluated, we could ask how the results vary with number tested, sampling smaller sets at random from the 1521. Hit rates and affinities were highly variable when we only sampled dozens of molecules, and it was only when we included several hundred molecules that results converged. As docking scores improved, so too did the likelihood of a molecule binding; hit rates improved steadily with docking score, as did affinities. This also appeared true on re-analysis of large-scale results against the {sigma}2 and dopamine D4 receptors. It may be that as the scale of both the virtual libraries and their testing grows, not only are better ligands found but so too does our ability to rank them.
]]></description>
<dc:creator>Liu, F.</dc:creator>
<dc:creator>Mailhot, O.</dc:creator>
<dc:creator>Glenn, I. S.</dc:creator>
<dc:creator>Vigneron, S. F.</dc:creator>
<dc:creator>Bassim, V.</dc:creator>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Valencia, K. F.</dc:creator>
<dc:creator>Smith, M. S.</dc:creator>
<dc:creator>Radchenko, D. S.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Moroz, Y. S.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2024-07-11</dc:date>
<dc:identifier>doi:10.1101/2024.07.08.602536</dc:identifier>
<dc:title><![CDATA[The impact of Library Size and Scale of Testing on Virtual Screening]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.08.602542v1?rss=1">
<title>
<![CDATA[
The nuclear periphery confers repression on H3K9me2-marked genes and transposons to shape cell fate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.08.602542v1?rss=1</link>
<description><![CDATA[
Heterochromatic loci marked by histone H3 lysine 9 dimethylation (H3K9me2) are enriched at the nuclear periphery in metazoans, but the effect of spatial position on heterochromatin function has not been defined. Here, we remove three nuclear lamins and lamin B receptor (LBR) in mouse embryonic stem cells (mESCs) and show that heterochromatin detaches from the nuclear periphery. Mutant mESCs sustain naive pluripotency and maintain H3K9me2 across the genome but cannot repress H3K9me2-marked genes or transposons. Further, mutant cells fail to differentiate into epiblast-like cells (EpiLCs), a transition that requires the expansion of H3K9me2 across the genome. Mutant EpiLCs can silence naive pluripotency genes and activate epiblast-stage genes. However, H3K9me2 cannot repress markers of alternative fates, including primitive endoderm. We conclude that the nuclear periphery controls the spatial position, dynamic remodeling, and repressive capacity of H3K9me2-marked heterochromatin to shape cell fate decisions.
]]></description>
<dc:creator>Marin, H.</dc:creator>
<dc:creator>Simental, E.</dc:creator>
<dc:creator>Allen, C.</dc:creator>
<dc:creator>Martin, E.</dc:creator>
<dc:creator>Panning, B.</dc:creator>
<dc:creator>Al-Sady, B.</dc:creator>
<dc:creator>Buchwalter, A.</dc:creator>
<dc:date>2024-07-11</dc:date>
<dc:identifier>doi:10.1101/2024.07.08.602542</dc:identifier>
<dc:title><![CDATA[The nuclear periphery confers repression on H3K9me2-marked genes and transposons to shape cell fate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.08.602602v1?rss=1">
<title>
<![CDATA[
Acute rapamycin treatment reveals novel mechanisms of behavioral, physiological, and functional dysfunction in a maternal inflammation mouse model of autism and sensory over-responsivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.08.602602v1?rss=1</link>
<description><![CDATA[
Maternal inflammatory response (MIR) during early gestation in mice induces a cascade of physiological and behavioral changes that have been associated with autism spectrum disorder (ASD). In a prior study and the current one, we find that mild MIR results in chronic systemic and brain inflammation, mTOR pathway activation, mild brain overgrowth followed by regionally specific volumetric changes, sensory processing dysregulation, and social and repetitive behavior abnormalities. Prior studies of rapamycin treatment in autism models have focused on chronic treatments that alter or prevent physical brain changes. Here, we have focused on the acute effects of rapamycin to uncover novel mechanisms of dysfunction related to mTOR pathway signaling. We find that within 2 hours, rapamycin treatment could rapidly rescue neuronal hyper-excitability, seizure susceptibility, functional network connectivity and brain community structure, repetitive behaviors, and sensory over-responsivity in adult offspring with persistent mild brain overgrowth. These CNS-mediated effects are also associated with alteration of the expression of several ASD-, ion channel-, and epilepsy-associated genes in the same time frame. Reduction of microglia with CSF1R inhibitors or inhibition of NADPH oxidase in young animals reduces the development of some of the behavioral phenotypes, but neither is as effective as acute mTOR inhibition. Our findings indicate that mTOR dysregulation in MIR offspring is a key contributor to various levels of brain dysfunction. However, we demonstrate that the adult MIR brain is also amenable to rapid normalization of these functional changes which results in the rescue of both core and comorbid ASD-like behaviors in adult animals without requiring long-term physical alterations to the brain. Restoring excitatory/inhibitory imbalance and sensory functional network modularity may therefore be important targets for therapeutically addressing both primary sensory and compensatory repetitive behavior phenotypes.
]]></description>
<dc:creator>Le Belle, J. E.</dc:creator>
<dc:creator>Condro, M.</dc:creator>
<dc:creator>Cepeda, C.</dc:creator>
<dc:creator>Oikonomou, K. D.</dc:creator>
<dc:creator>Tessema, K.</dc:creator>
<dc:creator>Dudley, L.</dc:creator>
<dc:creator>Schoenfield, J.</dc:creator>
<dc:creator>Kawaguchi, R.</dc:creator>
<dc:creator>Geschwind, D.</dc:creator>
<dc:creator>Silva, A. J.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:creator>Harris, N. G.</dc:creator>
<dc:creator>Kornblum, H. I.</dc:creator>
<dc:date>2024-07-11</dc:date>
<dc:identifier>doi:10.1101/2024.07.08.602602</dc:identifier>
<dc:title><![CDATA[Acute rapamycin treatment reveals novel mechanisms of behavioral, physiological, and functional dysfunction in a maternal inflammation mouse model of autism and sensory over-responsivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.09.602666v1?rss=1">
<title>
<![CDATA[
GATA6 regulates WNT and BMP programs to pattern precardiac mesoderm during the earliest stages of human cardiogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.09.602666v1?rss=1</link>
<description><![CDATA[
Haploinsufficiency for GATA6 is associated with congenital heart disease (CHD) with variable comorbidity of pancreatic or diaphragm defects, although the etiology of disease is not well understood. Here, we used cardiac directed differentiation from human embryonic stem cells (hESCs) as a platform to study GATA6 function during early cardiogenesis. GATA6 loss-of-function hESCs had a profound impairment in cardiac progenitor cell (CPC) specification and cardiomyocyte (CM) generation due to early defects during the mesendoderm and lateral mesoderm patterning stages. Profiling by RNA-seq and CUT&RUN identified genes of the WNT and BMP programs regulated by GATA6 during early mesoderm patterning. Furthermore, interactome analysis detected GATA6 binding with developmental transcription factors and chromatin remodelers suggesting cooperative regulation of cardiac lineage gene accessibility. We show that modulating WNT and BMP inputs during the first 48 hours of cardiac differentiation is sufficient to partially rescue CPC and CM defects in GATA6 heterozygous and homozygous mutant hESCs. This study provides evidence of the regulatory functions for GATA6 directing human precardiac mesoderm patterning during the earliest stages of cardiogenesis to further our understanding of haploinsufficiency causing CHD and the co-occurrence of cardiac and other organ defects caused by human GATA6 mutations.
]]></description>
<dc:creator>Bisson, J. A.</dc:creator>
<dc:creator>Gordillo, M.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>de Silva, N.</dc:creator>
<dc:creator>Yang, E.</dc:creator>
<dc:creator>Banks, K. M.</dc:creator>
<dc:creator>Shi, Z.-D.</dc:creator>
<dc:creator>Lee, K.</dc:creator>
<dc:creator>Yang, D.</dc:creator>
<dc:creator>Chung, W. K.</dc:creator>
<dc:creator>Huangfu, D.</dc:creator>
<dc:creator>Evans, T.</dc:creator>
<dc:date>2024-07-11</dc:date>
<dc:identifier>doi:10.1101/2024.07.09.602666</dc:identifier>
<dc:title><![CDATA[GATA6 regulates WNT and BMP programs to pattern precardiac mesoderm during the earliest stages of human cardiogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.11.601367v1?rss=1">
<title>
<![CDATA[
Basal p53 maintains a distinct transcriptional program from irradiated p53 in tissue, including tumor suppressors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.11.601367v1?rss=1</link>
<description><![CDATA[
The significance of p53s primary and secondary tumor suppressor programs cannot be overstated. A context- and stress-dependent transcription factor, p53 accumulates to mount its most well-characterized programs in response to a variety of stressors, most notably DNA damage. As cells and tissues never exist in a complete absence of stress, a small amount of p53 exists in cells under physiologic stress, detectable by chromatin immunoprecipitation and sequencing, termed basal p53. Recently, we and others have shown that basal p53 is sufficient to regulate tumor suppressor function. Furthermore, others have suggested the possibility that p53 accumulation in response to experimental stress may be dispensable for its tumor suppression. We previously showed basal p53 occupancy and regulation of known tumor suppressor genes, including PTEN and PHLDA3, in non-transformed breast cells, but this study was limited by experimental stress inherent to cell culture. Given the lack of global characterization of the basal p53 landscape under non-malignant physiologic stress in vivo, we utilized a multi-omics approach to define the murine basal p53 epigenome and its transcriptional program in various normal murine tissues. In this study, we observed basal p53 binding to cis regions of multiple tumor suppressor genes in different tissues, of which some showed p53-dependent regulation of their expression, including Phlda3, Bbc3, Xaf1, and itself. Furthermore, the vast majority of basal p53 target genes were not induced upon irradiation, suggesting basal p53 operates a transcriptional program that is largely distinct from its DNA damage response. Similarly, the basal p53 target gene repertoire is unique to each tissue type.
]]></description>
<dc:creator>Zhou, R. W.</dc:creator>
<dc:creator>Manfredi, J.</dc:creator>
<dc:creator>Silverman, L.</dc:creator>
<dc:creator>Parsons, R. E.</dc:creator>
<dc:date>2024-07-11</dc:date>
<dc:identifier>doi:10.1101/2024.07.11.601367</dc:identifier>
<dc:title><![CDATA[Basal p53 maintains a distinct transcriptional program from irradiated p53 in tissue, including tumor suppressors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.11.603114v1?rss=1">
<title>
<![CDATA[
Localized in vivo gene editing of murine cancer-associated fibroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.11.603114v1?rss=1</link>
<description><![CDATA[
Fibroblasts play critical roles in regulating cellular relationships during tissue homeostasis, immunity, and tumor biology at multiple sites. However, tools to perturb fibroblasts at just one site in vivo are limited, restricting our understanding of how these cellular relationships develop on a local level. We optimized local gene editing of fibroblasts in multiple mouse tumor models to investigate how locally restricted fibroblast perturbations affect the cellular tumor microenvironment (TME). By knocking out surface receptors Osmr, Tgfbr2, or Il1r1 on cancer-associated fibroblasts (CAFs), we uncover that TGFBR2 signaling loss uniquely induces the emergence of a Col18a1hi CAF cell state that is distinct from previously described fibroblast states and is associated with worse survival in human PDAC patients. Further application of a local as well as combinatorial gene knockout technology in CAFs reveals a circuit in which these Col18a1hi CAFs reshape the TME by recruiting Siglec-Fhi neutrophils via Cxcl5 expression; and that the Col18a1hi CAF cell state is further dependent on TNFR1 and canonical Wnt signaling. Together, a fast, affordable, and modular engineering method is demonstrated, allowing discovery of a modified fibroblast identify, as well as the network details of a local inter-cellular circuitry in a tumor.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=138 SRC="FIGDIR/small/603114v2_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@f375c5org.highwire.dtl.DTLVardef@1b8fad9org.highwire.dtl.DTLVardef@15cfd0aorg.highwire.dtl.DTLVardef@1e8dacc_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Kuhn, N. F.</dc:creator>
<dc:creator>Zaleta-Linares, I.</dc:creator>
<dc:creator>Nyberg, W. A.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2024-07-16</dc:date>
<dc:identifier>doi:10.1101/2024.07.11.603114</dc:identifier>
<dc:title><![CDATA[Localized in vivo gene editing of murine cancer-associated fibroblasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.14.603391v1?rss=1">
<title>
<![CDATA[
Peptide-enabled ribonucleoprotein delivery for CRISPR engineering (PERC) in primary human immune cells and hematopoietic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.14.603391v1?rss=1</link>
<description><![CDATA[
Peptide-enabled ribonucleoprotein delivery for CRISPR engineering (PERC) is a new approach for ex vivo genome editing of primary human cells. PERC uses a single amphiphilic peptide reagent to mediate intracellular delivery of the same pre-formed CRISPR ribonucleoprotein enzymes that are broadly used in research and therapeutics, resulting in high-efficiency editing of stimulated immune cells and cultured hematopoietic stem and progenitor cells (HSPCs). PERC facilitates nuclease-mediated gene knockout, precise transgene knock-in, and base editing. PERC involves mixing the CRISPR ribonucleoprotein enzyme with peptide and then incubating the formulation with cultured cells. For efficient transgene knock-in, adeno-associated virus (AAV) bearing homology-directed repair template DNA may be included. In contrast to electroporation, PERC is appealing as it requires no dedicated hardware and has less impact on cell phenotype and viability. Due to the gentle nature of PERC, delivery can be performed multiple times without substantial impact to cell health or phenotype. Here we report methods for improved PERC-mediated editing of T cells as well as novel methods for PERC-mediated editing of HSPCs, including knockout and precise knock-in. Editing efficiencies can surpass 90% using either Cas9 or Cas12a in primary T cells or HSPCs. Because PERC calls for only three readily available reagents - protein, RNA, and peptide - and does not require dedicated hardware for any step, PERC demands no special expertise and is exceptionally straightforward to adopt. The inherent compatibility of PERC with established cell engineering pipelines makes this approach appealing for rapid deployment in research and clinical settings.
]]></description>
<dc:creator>Sahu, S.</dc:creator>
<dc:creator>Castro, M.</dc:creator>
<dc:creator>Muldoon, J. J.</dc:creator>
<dc:creator>Asija, K.</dc:creator>
<dc:creator>Wyman, S. K.</dc:creator>
<dc:creator>Krishnappa, N.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Nguyen, D. N.</dc:creator>
<dc:creator>Wilson, R. C.</dc:creator>
<dc:date>2024-07-16</dc:date>
<dc:identifier>doi:10.1101/2024.07.14.603391</dc:identifier>
<dc:title><![CDATA[Peptide-enabled ribonucleoprotein delivery for CRISPR engineering (PERC) in primary human immune cells and hematopoietic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.15.603003v1?rss=1">
<title>
<![CDATA[
Sst- and Vip-Cre mouse lines without age-related hearing loss 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.15.603003v1?rss=1</link>
<description><![CDATA[
GABAergic interneurons, including somatostatin (SST) and vasoactive intestinal peptide (VIP) positive cells, play a crucial role in cortical circuit processing. Cre recombinase-mediated manipulation of these interneurons is facilitated by commercially available knock-in mouse strains such as Sst-IRES-Cre (Sst-Cre) and Vip-IRES-Cre (Vip-Cre). However, these strains are troublesome for hearing research because they are only available on the C57BL/6 genetic background, which suffer from early onset age-related hearing loss (AHL) due to a mutation of the Cdh23 gene. To overcome this limitation, we backcrossed Sst-Cre and Vip-Cre mice to CBA mice to create normal-hearing offspring with the desired Cre transgenes. We confirmed that in these "CBA Cre" lines, Cre drives appropriate expression of Cre-dependent genes, by crossing CBA Cre mice to Ai14 reporter mice. To assess the hearing capabilities of the CBA Cre mice, we measured auditory brainstem responses (ABRs) using clicks and tones. CBA Cre mice showed significantly lower ABR thresholds compared to C57 control mice at 3, 6, 9, and 12 months. In conclusion, our study successfully generated Sst-Cre and Vip-Cre mouse lines on the CBA background that will be valuable tools for investigating the roles of SST and VIP positive interneurons without the confounding effects of age-related hearing loss.
]]></description>
<dc:creator>Foss, C. T.</dc:creator>
<dc:creator>Olsen, T.</dc:creator>
<dc:creator>Bigelow, J.</dc:creator>
<dc:creator>Hasenstaub, A. R.</dc:creator>
<dc:date>2024-07-18</dc:date>
<dc:identifier>doi:10.1101/2024.07.15.603003</dc:identifier>
<dc:title><![CDATA[Sst- and Vip-Cre mouse lines without age-related hearing loss]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.16.603579v1?rss=1">
<title>
<![CDATA[
Mapping kinase domain resistance mechanisms for the MET receptor tyrosine kinase via deep mutational scanning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.16.603579v1?rss=1</link>
<description><![CDATA[
Mutations in the kinase and juxtamembrane domains of the MET Receptor Tyrosine Kinase are responsible for oncogenesis in various cancers and can drive resistance to MET-directed treatments. Determining the most effective inhibitor for each mutational profile is a major challenge for MET-driven cancer treatment in precision medicine. Here, we used a deep mutational scan (DMS) of [~]5,764 MET kinase domain variants to profile the growth of each mutation against a panel of 11 inhibitors that are reported to target the MET kinase domain. We validate previously identified resistance mutations, pinpoint common resistance sites across type I, type II, and type I [1/2] inhibitors, unveil unique resistance and sensitizing mutations for each inhibitor, and verify non-cross-resistant sensitivities for type I and type II inhibitor pairs. We augment a protein language model with biophysical and chemical features to improve the predictive performance for inhibitor-treated datasets. Together, our study demonstrates a pooled experimental pipeline for identifying resistance mutations, provides a reference dictionary for mutations that are sensitized to specific therapies, and offers insights for future drug development.
]]></description>
<dc:creator>Estevam, G. O.</dc:creator>
<dc:creator>Linossi, E. M.</dc:creator>
<dc:creator>Rao, J.</dc:creator>
<dc:creator>Macdonald, C. B.</dc:creator>
<dc:creator>Ravikumar, A.</dc:creator>
<dc:creator>Chrispens, K. M.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:creator>Coyote-Maestas, W.</dc:creator>
<dc:creator>Pimentel, H.</dc:creator>
<dc:creator>Collisson, E. A.</dc:creator>
<dc:creator>Jura, N.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2024-07-18</dc:date>
<dc:identifier>doi:10.1101/2024.07.16.603579</dc:identifier>
<dc:title><![CDATA[Mapping kinase domain resistance mechanisms for the MET receptor tyrosine kinase via deep mutational scanning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.17.603777v1?rss=1">
<title>
<![CDATA[
PATJ regulates cell stress responses and vascular remodeling post-stroke 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.17.603777v1?rss=1</link>
<description><![CDATA[
PALS1-associated tight junction (PATJ) protein is linked to metabolic disease and stroke in human genetic studies. Despite the recognized role of PATJ in cell polarization, its specific functions in metabolic disease and ischemic stroke recovery remain largely unexplored. Using a mouse model of stroke, we found post-ischemic stroke duration-dependent increase of PATJ abundance in endothelial cells. PATJ knock-out (KO) HEK293 cells generated by CRISPR-Cas9 suggest roles for PATJ in cell proliferation, migration, mitochondrial stress response, and interactions with the Yes-associated protein (YAP)-1 signaling pathway. Notably, PATJ deletion altered YAP1 nuclear translocation. PATJ KO cells demonstrated extensive transcriptional reprograming based on RNA sequencing analysis. Crucially, we identified dysregulation in genes central to vascular development, stress response, and metabolism, including RUNX1, HEY1, NUPR1, and HK2. These insights offer a new understanding of PATJs complex regulatory functions within cellular and vascular physiology and help lay the groundwork for therapeutic strategies targeting endothelial PATJ-mediated pathways for stroke rehabilitation and neurovascular repair.
]]></description>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Jiang, W.</dc:creator>
<dc:creator>Arkelius, K.</dc:creator>
<dc:creator>Swanson, R. A.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:creator>Singhal, N. S.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.17.603777</dc:identifier>
<dc:title><![CDATA[PATJ regulates cell stress responses and vascular remodeling post-stroke]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.19.603617v1?rss=1">
<title>
<![CDATA[
A Novel Whole Tissue Explant Model of Hidradenitis Suppurativa 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.19.603617v1?rss=1</link>
<description><![CDATA[
Hidradenitis Suppurativa (HS) is a relatively common and highly morbid inflammatory skin disease. Due to our relatively limited understanding of HSs pathogenesis, there are currently insufficient treatment options available, and many patients medical needs are not being met. This is partly due to a scarcity of ex vivo human assays and animal models that accurately recapitulate the disease. To address this deficit, we have developed a whole-tissue explant model of HS to examine its pathogenic mechanisms and the efficacy of potential treatments within intact human tissue. We measured cytokine protein and RNA within whole tissue maintained in an agar-media solution, finding that IL-6 and IL-8 concentrations trended upwards in both HS explants and healthy controls, while IL-17A, IL-1{beta}, and TNF- exhibited increases in HS tissue alone. We also show that the explants were responsive to treatment with both dexamethasone and IL-2. Not only do our results show that this model effectively delivers treatments throughout the explants, but they also elucidate which cytokines are related to the explant process regardless of tissue state and which are related to HS tissue specifically, laying the groundwork for future implementations of this model.
]]></description>
<dc:creator>Leboit, P. E.</dc:creator>
<dc:creator>Patel, D. U.</dc:creator>
<dc:creator>Cohen, J. N.</dc:creator>
<dc:creator>Moss, M. I.</dc:creator>
<dc:creator>Naik, H. B.</dc:creator>
<dc:creator>Yates, A. E.</dc:creator>
<dc:creator>Harris, H. W.</dc:creator>
<dc:creator>Klufas, D. M.</dc:creator>
<dc:creator>Kim, E. A.</dc:creator>
<dc:creator>Neuhaus, I. M.</dc:creator>
<dc:creator>Hansen, S. L.</dc:creator>
<dc:creator>Kyle, R. L.</dc:creator>
<dc:creator>Kelly, M.</dc:creator>
<dc:creator>Rosenblum, M. D.</dc:creator>
<dc:creator>Lowe, M. M.</dc:creator>
<dc:date>2024-07-23</dc:date>
<dc:identifier>doi:10.1101/2024.07.19.603617</dc:identifier>
<dc:title><![CDATA[A Novel Whole Tissue Explant Model of Hidradenitis Suppurativa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.22.602974v1?rss=1">
<title>
<![CDATA[
High-yield production of recombinant human myelin oligodendrocyte glycoprotein in SHuffle bacteria without a refolding step 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.22.602974v1?rss=1</link>
<description><![CDATA[
1.1Experimental autoimmune encephalomyelitis (EAE) is a model for central nervous system (CNS) autoimmune demyelinating diseases such as multiple sclerosis (MS) and MOG antibody-associated disease (MOGAD). Immunization with the extracellular domain of recombinant human MOG (rhMOG), which contains pathogenic antibody and T cell epitopes, induces B cell-dependent EAE for studies in mice. However, these studies have been hampered by rhMOG availability due to its insolubility when overexpressed in bacterial cells, and the requirement for inefficient denaturation and refolding. Here, we describe a new protocol for the high-yield production of soluble rhMOG in SHuffle cells, a commercially available E. coli strain engineered to facilitate disulfide bond formation in the cytoplasm. SHuffle cells can produce a soluble fraction of rhMOG yielding >100 mg/L. Analytical size exclusion chromatography multi-angle light scattering (SEC-MALS) and differential scanning fluorimetry of purified rhMOG reveals a homogeneous monomer with a high melting temperature, indicative of a well-folded protein. An in vitro proliferation assay establishes that purified rhMOG can be processed and recognized by T cells expressing a T cell receptor (TCR) specific for the immunodominant MOG35-55 peptide epitope. Lastly, immunization of wild-type, but not B cell deficient, mice with rhMOG resulted in robust induction of EAE, indicating a B cell-dependent induction. Our SHuffle cell method greatly simplifies rhMOG production by combining the high yield and speed of bacterial cell expression with enhanced disulfide bond formation and folding, which will enable further investigation of B cell-dependent EAE and expand human research of MOG in CNS demyelinating diseases.
]]></description>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Sagan, S. A.</dc:creator>
<dc:creator>Moseley, C. E.</dc:creator>
<dc:creator>Zamvil, S. S.</dc:creator>
<dc:creator>Pak, J. E.</dc:creator>
<dc:date>2024-07-23</dc:date>
<dc:identifier>doi:10.1101/2024.07.22.602974</dc:identifier>
<dc:title><![CDATA[High-yield production of recombinant human myelin oligodendrocyte glycoprotein in SHuffle bacteria without a refolding step]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.21.601827v1?rss=1">
<title>
<![CDATA[
Therapeutic DBS for OCD Suppresses the Default Mode Network 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.21.601827v1?rss=1</link>
<description><![CDATA[
BackgroundDeep brain stimulation (DBS) of the anterior limb of the internal capsule (ALIC) is an emerging treatment for severe, refractory obsessive-compulsive disorder (OCD). The therapeutic effects of DBS are hypothesized to be mediated by direct modulation of a distributed cortico-striato-thalmo-cortical network underlying OCD symptoms. However, the exact underlying mechanism by which DBS exerts its therapeutic effects still remains unclear.

MethodIn five participants receiving DBS for severe, refractory OCD (3 responders, 2 non-responders), we conducted a DBS On/Off cycling paradigm during the acquisition of functional MRI to determine the network effects of stimulation across a variety of bipolar configurations. We also performed tractography using diffusion-weighted imaging (DWI) to relate the functional impact of DBS to the underlying structural connectivity between active stimulation contacts and functional brain networks.

ResultsWe found that therapeutic DBS had a distributed effect, suppressing BOLD activity within regions such as the orbitofrontal cortex, dorsomedial prefrontal cortex, and subthalamic nuclei compared to non-therapeutic configurations. Many of the regions suppressed by therapeutic DBS were components of the default mode network (DMN). Moreover, the estimated stimulation field from the therapeutic configurations exhibited significant structural connectivity to core nodes of the DMN.

ConclusionsTherapeutic DBS for OCD suppresses BOLD activity within a distributed set of regions within the DMN relative to non-therapeutic configurations. We propose that these effects may be mediated by interruption of communication through structural white matter connections surrounding the DBS active contacts.
]]></description>
<dc:creator>Slepneva, N.</dc:creator>
<dc:creator>Basich-Pease, G.</dc:creator>
<dc:creator>Reid, L.</dc:creator>
<dc:creator>Frank, A. C.</dc:creator>
<dc:creator>Norbu, T.</dc:creator>
<dc:creator>Krystal, A. D.</dc:creator>
<dc:creator>Sugrue, L. P.</dc:creator>
<dc:creator>Motzkin, J. C.</dc:creator>
<dc:creator>Larson, P. S.</dc:creator>
<dc:creator>Starr, P. A.</dc:creator>
<dc:creator>Morrison, M. A.</dc:creator>
<dc:creator>Lee, A. M.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.21.601827</dc:identifier>
<dc:title><![CDATA[Therapeutic DBS for OCD Suppresses the Default Mode Network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.23.604673v1?rss=1">
<title>
<![CDATA[
Genetic evolution of keratinocytes to cutaneous squamous cell carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.23.604673v1?rss=1</link>
<description><![CDATA[
We performed multi-omic profiling of epidermal keratinocytes, precancerous actinic keratoses, and squamous cell carcinomas to understand the molecular transitions during skin carcinogenesis. Single-cell mutational analyses of normal skin cells showed that most keratinocytes have remarkably low mutation burdens, despite decades of sun exposure, however keratinocytes with TP53 or NOTCH1 mutations had substantially higher mutation burdens. These observations suggest that wild-type keratinocytes (i.e. without pathogenic mutations) are able to withstand high dosages of cumulative UV radiation, but certain pathogenic mutations break these adaptive mechanisms, priming keratinocytes for transformation by increasing their mutation rate. Mutational profiling of squamous cell carcinomas adjacent to actinic keratoses revealed TERT promoter and CDKN2A mutations emerging in actinic keratoses, whereas additional mutations inactivating ARID2 and activating the MAPK-pathway delineated the transition to squamous cell carcinomas. Surprisingly, actinic keratoses were often not related to their neighboring squamous cell carcinoma, indicating that collisions of unrelated neoplasms are common in the skin. Spatial variation in gene expression patterns was common in both tumor and immune cells, with high expression of checkpoint molecules at the invasive front of tumors. In conclusion, this study catalogues the key events during the evolution of cutaneous squamous cell carcinoma.
]]></description>
<dc:creator>Tandukar, B.</dc:creator>
<dc:creator>Deivendran, D.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Cruz-Pacheco, N.</dc:creator>
<dc:creator>Sharma, H.</dc:creator>
<dc:creator>Xu, A.</dc:creator>
<dc:creator>Bandari, A. K.</dc:creator>
<dc:creator>Chen, D.</dc:creator>
<dc:creator>George, C.</dc:creator>
<dc:creator>Marty, A.</dc:creator>
<dc:creator>Cho, R. J.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Saylor, D.</dc:creator>
<dc:creator>Gerami, P.</dc:creator>
<dc:creator>Arron, S. T.</dc:creator>
<dc:creator>Bastian, B. C.</dc:creator>
<dc:creator>Shain, A. H.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.23.604673</dc:identifier>
<dc:title><![CDATA[Genetic evolution of keratinocytes to cutaneous squamous cell carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.23.604809v1?rss=1">
<title>
<![CDATA[
Mechanism-guided engineering of a minimal biological particle for genome editing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.23.604809v1?rss=1</link>
<description><![CDATA[
The widespread application of genome editing to treat or even cure disease requires the delivery of genome editors into the nucleus of target cells. Enveloped Delivery Vehicles (EDVs) are engineered virally-derived particles capable of packaging and delivering CRISPR-Cas9 ribonucleoproteins (RNPs). However, the presence of lentiviral genome encapsulation and replication components in EDVs has obscured the underlying delivery mechanism and precluded particle optimization. Here we show that Cas9 RNP nuclear delivery is independent of the native lentiviral capsid structure. Instead, EDV-mediated genome editing activity corresponds directly to the number of nuclear localization sequences on the Cas9 enzyme. EDV structural analysis using cryo-electron tomography and small molecule inhibitors guided the removal of [~]80% of viral residues, creating a minimal EDV (miniEDV) that retains full RNP delivery capability. MiniEDVs are 25% smaller yet package equivalent amounts of Cas9 RNPs relative to the original EDVs, and demonstrated increased editing in cell lines and therapeutically-relevant primary human T cells. These results show that virally-derived particles can be streamlined to create efficacious genome editing delivery vehicles that could simplify production and manufacturing.

SIGNIFICANCE STATEMENTOur results highlight the importance of understanding how virally-derived particles function to eliminate unnecessary viral proteins and create more efficacious and easier-to-produce delivery vehicles for therapeutic genome editing.
]]></description>
<dc:creator>Ngo, W.</dc:creator>
<dc:creator>Peukes, J. T.</dc:creator>
<dc:creator>Baldwin, A.</dc:creator>
<dc:creator>Xue, Z. W.</dc:creator>
<dc:creator>Hwang, S.</dc:creator>
<dc:creator>Stickels, R. R.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Satpathy, A.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:creator>Schekman, R.</dc:creator>
<dc:creator>Nogales, E.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.23.604809</dc:identifier>
<dc:title><![CDATA[Mechanism-guided engineering of a minimal biological particle for genome editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.23.604849v1?rss=1">
<title>
<![CDATA[
Genetic Architecture and Analysis Practices of Circulating Metabolites in the NHLBI Trans-Omics for Precision Medicine (TOPMed) Program 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.23.604849v1?rss=1</link>
<description><![CDATA[
Circulating metabolite levels partly reflect the state of human health and diseases and can be impacted by genetic determinants. Hundreds of loci associated with circulating metabolites have been identified; however, most findings focus on predominantly European ancestry or single-study analyses. Leveraging the rich metabolomics resources generated by the NHLBI Trans-Omics for Precision Medicine (TOPMed) Program, we harmonized and accessibly cataloged 1,729 circulating metabolites among 25,058 ancestrally diverse samples. We provided a set of reasonable strategies for outlier and imputation handling to process metabolite data. Following the practical analysis framework, we further performed a genome-wide association analysis on 1,135 selected metabolites using whole genome sequencing data from 16,359 individuals passing the quality control filters, and discovered 1,778 independent loci associated with 667 metabolites. Among 108 novel locus-metabolite pairs, we detected not only novel loci within previously implicated metabolite associated genes but also novel genes (such as GAB3 and VSIG4 located in the X chromosome) that have putative roles in metabolic regulation. In the sex-stratified analysis, we revealed 85 independent locus-metabolite pairs with evidence of sexual dimorphism, including well-known metabolic genes such as FADS2, D2HGDH, SUGP1, UTG2B17, strongly supporting the importance of exploring sex difference in the human metabolome. Taken together, our study depicted the genetic contribution to circulating metabolite levels, providing additional insight into the understanding of human health.
]]></description>
<dc:creator>Wang, N.</dc:creator>
<dc:creator>Ockerman, F. P.</dc:creator>
<dc:creator>Zhou, L. Y.</dc:creator>
<dc:creator>Grove, M. L.</dc:creator>
<dc:creator>Alkis, T.</dc:creator>
<dc:creator>Barnard, J.</dc:creator>
<dc:creator>Bowler, R. P.</dc:creator>
<dc:creator>Clish, C. B.</dc:creator>
<dc:creator>Chung, S.</dc:creator>
<dc:creator>Drzymalla, E.</dc:creator>
<dc:creator>Evans, A. M.</dc:creator>
<dc:creator>Franceschini, N.</dc:creator>
<dc:creator>Gerszten, R. E.</dc:creator>
<dc:creator>Gillman, M. G.</dc:creator>
<dc:creator>Hutton, S. R.</dc:creator>
<dc:creator>Kelly, R. S.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Larson, M. G.</dc:creator>
<dc:creator>Lasky-Su, J. A.</dc:creator>
<dc:creator>Meyers, D. A.</dc:creator>
<dc:creator>Woodruff, P. G.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Rich, S. S.</dc:creator>
<dc:creator>Rotter, J.</dc:creator>
<dc:creator>Silverman, E. K.</dc:creator>
<dc:creator>Ramachandran, V. S.</dc:creator>
<dc:creator>Weiss, S. T.</dc:creator>
<dc:creator>Wong, K. E.</dc:creator>
<dc:creator>Wood, A. C.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>NHLBI Trans-Omics for Precision Medicine (TOPMed) Consortium,</dc:creator>
<dc:creator>Yarden, R.</dc:creator>
<dc:creator>Blackwell, T.</dc:creator>
<dc:creator>Smith, A. V.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Raffield, L. M.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.23.604849</dc:identifier>
<dc:title><![CDATA[Genetic Architecture and Analysis Practices of Circulating Metabolites in the NHLBI Trans-Omics for Precision Medicine (TOPMed) Program]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.24.604997v1?rss=1">
<title>
<![CDATA[
Comparative Analysis of Glycoproteomic Software Using a Tailored Glycan Database 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.24.604997v1?rss=1</link>
<description><![CDATA[
Glycoproteomics is a rapidly developing field, and data analysis has been stimulated by several technological innovations. As a result, there are many software tools from which to choose; and each comes with unique features that can be difficult to compare. This work presents a head-to-head comparison of five modern analytical software: Byonic, Protein Prospector, MSFraggerGlyco, pGlyco3, and GlycoDecipher. To enable a meaningful comparison, parameter variables were minimized. One potential confounding variable is the glycan database that informs glycoproteomic searches. We performed glycomic profiling of the samples and used the output to construct matched glycan databases for each software. Up to 19,000 glycopeptide spectra were identified across three replicates of wild-type SH-SY5Y cells. There was substantial overlap among most software for glycoproteins identified, locations of glycosites, and glycans, although Byonic reported a suspiciously large number of glycoproteins and glycosites of questionable reliability. We show that Protein Prospector identified the most glycopeptide spectrum matches with high agreement to known glycosites in UniProt. Overall, our results indicate that glycoproteomic searches should involve more than one software to generate confidence. It may be useful to consider software with peptide-first approaches and with glycan-first approaches.
]]></description>
<dc:creator>Hogan, R. A.</dc:creator>
<dc:creator>Pepi, L. E.</dc:creator>
<dc:creator>Riley, N. M.</dc:creator>
<dc:creator>Chalkley, R. J.</dc:creator>
<dc:date>2024-07-25</dc:date>
<dc:identifier>doi:10.1101/2024.07.24.604997</dc:identifier>
<dc:title><![CDATA[Comparative Analysis of Glycoproteomic Software Using a Tailored Glycan Database]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.26.605328v1?rss=1">
<title>
<![CDATA[
A second X chromosome improves cognition in aging male and female mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.26.605328v1?rss=1</link>
<description><![CDATA[
Women show resilience to cognitive aging, in the absence of dementia, in many populations. To dissect sex differences, we utilized the FCG and XY*mouse models. Female gonads and sex chromosomes improved cognition in aging mice of both sexes. Further, presence of a second X in male and female mice conferred cognitive resilience while its absence in females blocked it. In the hippocampal proteome of aging female mice, the second X increased proteins involved in synaptogenesis signaling - a potential pathway to improved cognition.
]]></description>
<dc:creator>Marino, F.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Merrihew, G. E.</dc:creator>
<dc:creator>MacCoss, M. J.</dc:creator>
<dc:creator>Dubal, D.</dc:creator>
<dc:date>2024-07-26</dc:date>
<dc:identifier>doi:10.1101/2024.07.26.605328</dc:identifier>
<dc:title><![CDATA[A second X chromosome improves cognition in aging male and female mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.26.604313v1?rss=1">
<title>
<![CDATA[
Proteomic Profiling Reveals Age-Related Changes in Transporter Proteins in the Human Blood-Brain Barrier 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.26.604313v1?rss=1</link>
<description><![CDATA[
The Blood-Brain Barrier (BBB) is a crucial, selective barrier that regulates the entry of molecules including nutrients, environmental toxins, and therapeutic medications into the brain. This function relies heavily on brain endothelial cell proteins, particularly transporters and tight junction proteins. The BBB continues to develop postnatally, adapting its selective barrier function across different developmental phases, and alters with aging and disease. Here we present a global proteomics analysis focused on the ontogeny and aging of proteins in human brain microvessels (BMVs), predominantly composed of brain endothelial cells. Our proteomic profiling quantified 6,223 proteins and revealed possible age-related alteration in BBB permeability due to basement membrane component changes through the early developmental stage and age-dependent changes in transporter expression. Notable changes in expression levels were observed with development and age in nutrient transporters and transporters that play critical roles in drug disposition. This research 1) provides important information on the mechanisms that drive changes in the metabolic content of the brain with age and 2) enables the creation of physiologically based pharmacokinetic models for CNS drug distribution across different life stages.
]]></description>
<dc:creator>Zhou, X.</dc:creator>
<dc:creator>Azimi, M.</dc:creator>
<dc:creator>Handin, N.</dc:creator>
<dc:creator>Riselli, A.</dc:creator>
<dc:creator>Vora, B.</dc:creator>
<dc:creator>Chun, E.</dc:creator>
<dc:creator>Yee, S. W.</dc:creator>
<dc:creator>Artursson, P.</dc:creator>
<dc:creator>Giacomini, K. M.</dc:creator>
<dc:date>2024-07-27</dc:date>
<dc:identifier>doi:10.1101/2024.07.26.604313</dc:identifier>
<dc:title><![CDATA[Proteomic Profiling Reveals Age-Related Changes in Transporter Proteins in the Human Blood-Brain Barrier]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.29.605645v1?rss=1">
<title>
<![CDATA[
Senescent fibroblasts in the tumor stroma rewire lung cancer metabolism and plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.29.605645v1?rss=1</link>
<description><![CDATA[
Senescence has been demonstrated to either inhibit or promote tumorigenesis. Resolving this paradox requires spatial mapping and functional characterization of senescent cells in the native tumor niche. Here, we identified senescent p16Ink4a+ cancer-associated fibroblasts with a secretory phenotype that promotes fatty acid uptake and utilization by aggressive lung adenocarcinoma driven by Kras and p53 mutations. Furthermore, rewiring of lung cancer metabolism by p16Ink4a+ cancer- associated fibroblasts also altered tumor cell identity to a highly plastic/dedifferentiated state associated with progression in murine and human LUAD. Our ex vivo senolytic screening platform identified XL888, a HSP90 inhibitor, that cleared p16Ink4a+ cancer- associated fibroblasts in vivo. XL888 administration after establishment of advanced lung adenocarcinoma significantly reduced tumor burden concurrent with the loss of plastic tumor cells. Our study identified a druggable component of the tumor stroma that fulfills the metabolic requirement of tumor cells to acquire a more aggressive phenotype.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Nabora, S. R.</dc:creator>
<dc:creator>Woo, S.-H.</dc:creator>
<dc:creator>Goel, S.</dc:creator>
<dc:creator>Stratton, F.</dc:creator>
<dc:creator>Kuang, C.</dc:creator>
<dc:creator>Mansfield, A.</dc:creator>
<dc:creator>LaFave, L.</dc:creator>
<dc:creator>Peng, T.</dc:creator>
<dc:date>2024-07-30</dc:date>
<dc:identifier>doi:10.1101/2024.07.29.605645</dc:identifier>
<dc:title><![CDATA[Senescent fibroblasts in the tumor stroma rewire lung cancer metabolism and plasticity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.29.605664v1?rss=1">
<title>
<![CDATA[
Conservation, alteration, and redistribution of mammalian striatal interneurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.29.605664v1?rss=1</link>
<description><![CDATA[
Mammalian brains vary in size, structure, and function, but the extent to which evolutionarily novel cell types contribute to this variation remains unresolved1-4. Recent studies suggest there is a primate-specific population of striatal inhibitory interneurons, the TAC3 interneurons5. However, there has not yet been a detailed analysis of the spatial and phylogenetic distribution of this population. Here, we profile single cell gene expression in the developing pig (an ungulate) and ferret (a carnivore), representing 94 million years divergence from primates, and assign newborn inhibitory neurons to initial classes first specified during development6. We find that the initial class of TAC3 interneurons represents an ancestral striatal population that is also deployed towards the cortex in pig and ferret. In adult mouse, we uncover a rare population expressing Tac2, the ortholog of TAC3, in ventromedial striatum, prompting a reexamination of developing mouse striatal interneuron initial classes by targeted enrichment of their precursors. We conclude that the TAC3 interneuron initial class is conserved across Boreoeutherian mammals, with the mouse population representing Th striatal interneurons, a subset of which expresses Tac2. This study suggests that initial classes of telencephalic inhibitory neurons are largely conserved and that during evolution, neuronal types in the mammalian brain change through redistribution and fate refinement, rather than by derivation of novel precursors early in development.
]]></description>
<dc:creator>Corrigan, E. K.</dc:creator>
<dc:creator>DeBerardine, M.</dc:creator>
<dc:creator>Poddar, A.</dc:creator>
<dc:creator>Turrero Garcia, M.</dc:creator>
<dc:creator>Schmitz, M. T.</dc:creator>
<dc:creator>Harwell, C.</dc:creator>
<dc:creator>Paredes, M.</dc:creator>
<dc:creator>Krienen, F. M.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:date>2024-07-30</dc:date>
<dc:identifier>doi:10.1101/2024.07.29.605664</dc:identifier>
<dc:title><![CDATA[Conservation, alteration, and redistribution of mammalian striatal interneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.29.605671v1?rss=1">
<title>
<![CDATA[
Calcium phosphate nanoclusters modify periodontium remodeling and minimize orthodontic relapse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.29.605671v1?rss=1</link>
<description><![CDATA[
Orthodontic relapse is one of the most prevalent concerns of orthodontic therapy. Relapse results in patients teeth reverting towards their pretreatment positions, which increases the susceptibility to functional problems, dental disease, and substantially increases the financial burden for retreatment. This phenomenon is thought to be induced by rapid remodeling of the periodontal ligament (PDL) in the early stages and poor bone quality in the later stages. Current therapies, including fixed or removable retainers and fiberotomies, have limitations with patient compliance and invasiveness. Approaches using biocompatible biomaterials, such as calcium phosphate polymer-induced liquid precursors (PILP), is an ideal translational approach for minimizing orthodontic relapse. Here, post-orthodontic relapse is reduced after a single injection of high concentration PILP (HC-PILP) nanoclusters by altering PDL remodeling in the early stage of relapse and improving trabecular bone quality in the later phase. HC-PILP nanoclusters are achieved by using high molecular weight poly aspartic acid (PASP, 14 kDa) and poly acrylic acid (PAA, 450 kDa), which resulted in a stable solution of high calcium and phosphate concentrations without premature precipitation. In vitro results show that HC-PILP nanoclusters prevented collagen type-I mineralization, which is essential for the tooth-periodontal ligament (PDL)-bone interphase. In vivo experiments show that the PILP nanoclusters minimize relapse and improve the trabecular bone quality in the late stages of relapse. Interestingly, PILP nanoclusters also altered the remodeling of the PDL collagen during the early stages of relapse. Further in vitro experiments showed that PILP nanoclusters alter the fibrillogenesis of collagen type-I by impacting the protein secondary structure. These findings propose a novel approach for treating orthodontic relapse and provide additional insight into the PILP nanoclusters structure and properties on collagenous structure repair.
]]></description>
<dc:creator>Cuylear, D. L.</dc:creator>
<dc:creator>Fu, M. L.</dc:creator>
<dc:creator>Chau, J. C.</dc:creator>
<dc:creator>Kharbikar, B.</dc:creator>
<dc:creator>Kazakia, G.</dc:creator>
<dc:creator>Jheon, A.</dc:creator>
<dc:creator>Habelitz, S.</dc:creator>
<dc:creator>Kapila, S. D.</dc:creator>
<dc:creator>Desai, T. A.</dc:creator>
<dc:date>2024-07-30</dc:date>
<dc:identifier>doi:10.1101/2024.07.29.605671</dc:identifier>
<dc:title><![CDATA[Calcium phosphate nanoclusters modify periodontium remodeling and minimize orthodontic relapse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.30.602578v1?rss=1">
<title>
<![CDATA[
Ciliary biology intersects autism and congenital heart disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.30.602578v1?rss=1</link>
<description><![CDATA[
Autism spectrum disorder (ASD) commonly co-occurs with congenital heart disease (CHD), but the molecular mechanisms underlying this comorbidity remain unknown. Given that children with CHD come to clinical attention by the newborn period, understanding which CHD variants carry ASD risk could provide an opportunity to identify and treat individuals at high risk for developing ASD far before the typical age of diagnosis. Therefore, it is critical to delineate the subset of CHD genes most likely to increase the risk of ASD. However, to date there is relatively limited overlap between high confidence ASD and CHD genes, suggesting that alternative strategies for prioritizing CHD genes are necessary. Recent studies have shown that ASD gene perturbations commonly dysregulate neural progenitor cell (NPC) biology. Thus, we hypothesized that CHD genes that disrupt neurogenesis are more likely to carry risk for ASD. Hence, we performed an in vitro pooled CRISPR interference (CRISPRi) screen to identify CHD genes that disrupt NPC biology similarly to ASD genes. Overall, we identified 45 CHD genes that strongly impact proliferation and/or survival of NPCs. Moreover, we observed that a cluster of physically interacting ASD and CHD genes are enriched for ciliary biology. Studying seven of these genes with evidence of shared risk (CEP290, CHD4, KMT2E, NSD1, OFD1, RFX3, TAOK1), we observe that perturbation significantly impacts primary cilia formation in vitro. While in vivo investigation of TAOK1 reveals a previously unappreciated role for the gene in motile cilia formation and heart development, supporting its prediction as a CHD risk gene. Together, our findings highlight a set of CHD risk genes that may carry risk for ASD and underscore the role of cilia in shared ASD and CHD biology.
]]></description>
<dc:creator>Teerikorpi, N.</dc:creator>
<dc:creator>Lasser, M. C.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Kostyanovskaya, E.</dc:creator>
<dc:creator>Bader, E.</dc:creator>
<dc:creator>Sun, N.</dc:creator>
<dc:creator>Dea, J.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Willsey, A. J.</dc:creator>
<dc:creator>Willsey, H.</dc:creator>
<dc:date>2024-07-31</dc:date>
<dc:identifier>doi:10.1101/2024.07.30.602578</dc:identifier>
<dc:title><![CDATA[Ciliary biology intersects autism and congenital heart disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.01.605860v1?rss=1">
<title>
<![CDATA[
Using virtual patient cohorts to uncover immune response differences in cancer and immunosuppressed COVID-19 patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.01.605860v1?rss=1</link>
<description><![CDATA[
The COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) resulted in millions of deaths globally. Adults with immunosuppression (e.g., solid organ transplant recipients) and those undergoing active cancer treatments experience worse infections and more severe COVID-19. It is difficult to conduct clinical studies in these populations, resulting in a restricted amount of data that can be used to relate mechanisms of immune dysfunction to COVID-19 outcomes in these vulnerable groups. To study immune dynamics after infection with SARS-CoV-2 and to investigate drivers of COVID-19 severity in individuals with cancer and immunosuppression, we adapted our mathematical model of the immune response during COVID-19 and generated virtual patient cohorts of cancer and immunosuppressed patients. The cohorts of plausible patients recapitulated available longitudinal clinical data collected from patients in Montreal, Canada area hospitals. Our model predicted that both cancer and immunosuppressed virtual patients with severe COVID-19 had decreased CD8+ T cells, elevated interleukin-6 concentrations, and delayed type I interferon peaks compared to those with mild COVID-19 outcomes. Additionally, our results suggest that cancer patients experience higher viral loads (however, with no direct relation with severity), likely because of decreased initial neutrophil counts (i.e., neutropenia), a frequent toxic side effect of anti-cancer therapy. Furthermore, severe cancer and immunosuppressed virtual patients suffered a high degree of tissue damage associated with elevated neutrophils. Lastly, parameter values associated with monocyte recruitment by infected cells were found to be elevated in severe cancer and immunosuppressed patients with respect to the COVID-19 reference group. Together, our study highlights that dysfunction in type I interferon and CD8+ T cells are key drivers of immune dysregulation in COVID-19, particularly in cancer patients and immunosuppressed individuals.
]]></description>
<dc:creator>Gazeau, S.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Brunet-Ratnasingham, E.</dc:creator>
<dc:creator>Kaufmann, D. E.</dc:creator>
<dc:creator>Larochelle, C.</dc:creator>
<dc:creator>Morel, P. A.</dc:creator>
<dc:creator>Heffernan, J. M.</dc:creator>
<dc:creator>Davis, C. L.</dc:creator>
<dc:creator>Smith, A. M.</dc:creator>
<dc:creator>Jenner, A. L.</dc:creator>
<dc:creator>Craig, M.</dc:creator>
<dc:date>2024-08-02</dc:date>
<dc:identifier>doi:10.1101/2024.08.01.605860</dc:identifier>
<dc:title><![CDATA[Using virtual patient cohorts to uncover immune response differences in cancer and immunosuppressed COVID-19 patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.01.606248v1?rss=1">
<title>
<![CDATA[
Spatial 3D genome organization controls the activity of bivalent chromatin during human neurogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.01.606248v1?rss=1</link>
<description><![CDATA[
The nuclear genome is spatially organized into a three-dimensional (3D) architecture by physical association of large chromosomal domains with subnuclear compartments including the nuclear lamina at the radial periphery and nuclear speckles within the nucleoplasm1-5. However, how spatial genome architecture regulates human brain development has been overlooked owing to technical limitations. Here, we generate high-resolution maps of genomic interactions with the lamina and speckles in cells of the neurogenic lineage isolated from midgestational human cortex, uncovering an intimate association between subnuclear genome compartmentalization, chromatin state and transcription. During cortical neurogenesis, spatial genome organization is extensively remodeled, relocating hundreds of neuronal genes from the lamina to speckles including key neurodevelopmental genes bivalent for H3K27me3 and H3K4me3. At the lamina, bivalent genes have exceptionally low expression, and relocation to speckles enhances resolution of bivalent chromatin to H3K4me3 and increases transcription >7-fold. We further demonstrate that proximity to the nuclear periphery - not the presence of H3K27me3 - is the dominant factor in maintaining the lowly expressed, poised state of bivalent genes embedded in the lamina. In addition to uncovering a critical role of subnuclear genome compartmentalization in neurogenic transcriptional regulation, our results establish a new paradigm in which knowing the spatial location of a gene is necessary to understanding its epigenomic regulation.
]]></description>
<dc:creator>Ahanger, S. H.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Semenza, E. R.</dc:creator>
<dc:creator>Gil, E.</dc:creator>
<dc:creator>Cole, M. A.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Kriegstein, A. R.</dc:creator>
<dc:creator>Lim, D.</dc:creator>
<dc:date>2024-08-02</dc:date>
<dc:identifier>doi:10.1101/2024.08.01.606248</dc:identifier>
<dc:title><![CDATA[Spatial 3D genome organization controls the activity of bivalent chromatin during human neurogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.03.606273v1?rss=1">
<title>
<![CDATA[
Phylogeny, morphology, and behavior of the new ciliate species Stentor stipatus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.03.606273v1?rss=1</link>
<description><![CDATA[
BackgroundStentor, the genus of large trumpet-shaped ciliates, is well-known for its complex morphology and striking behaviors. Members of this genus are distributed throughout the world in a wide and diverse pool of freshwater ecosystems. Recently, the molecular phylogeny of Stentor has been explored through comparison of 18S small subunit (SSU) ribosomal DNA (rDNA) sequences, clarifying several previously mischaracterized species and species complexes. However, despite their wide distribution, to-date, only about a dozen species of Stentor have been described and verified by phylogenetic means.

ResultsHere, we introduce the discovery of a new species within genus Stentor: Stentor stipatus spec. nov., so named for their distinctive cytosolic dark pigmented granules which surround the macronucleus and are also present cortically alongside cortically-distributed green microalgae. We present morphological, phylogenetic, ecological, and behavioral characterizations of these cells. Phylogenetic analysis of S. stipatus spec. nov. by comparison of SSU rDNA sequence suggests it is a distinct species from its closest relative, S. amethystinus. We demonstrate that S. stipatus spec. nov. is capable of habituation in response to repeated mechanical stimulation. Further, S. stipatus spec. nov. exhibits strongly directed positive phototaxis, like its relative S. pyriformis, but with a distinct action spectrum from both S. coeruleus and S. pyriformis. Finally, S. stipatus phototaxis response strength varies in a consistent pattern throughout the day, providing evidence of potential circadian regulation.

ConclusionsThis work expands the current understanding of the ecological distribution of and behavioral features present within genus Stentor.
]]></description>
<dc:creator>Rajan, D. H.</dc:creator>
<dc:creator>Lee, B.</dc:creator>
<dc:creator>Albright, A.</dc:creator>
<dc:creator>Tang, E. J.</dc:creator>
<dc:creator>Maravillas, A.</dc:creator>
<dc:creator>Vargas, C.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:creator>Cortes, D.</dc:creator>
<dc:date>2024-08-04</dc:date>
<dc:identifier>doi:10.1101/2024.08.03.606273</dc:identifier>
<dc:title><![CDATA[Phylogeny, morphology, and behavior of the new ciliate species Stentor stipatus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.03.606488v1?rss=1">
<title>
<![CDATA[
CRISPR-Cas13d as a molecular tool to achieve targeted gene expression knockdown in chick embryos 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.03.606488v1?rss=1</link>
<description><![CDATA[
The chick embryo is a classical model system commonly used in developmental biology due to its amenability to gene perturbation experiments. Pairing this powerful model organism with cutting-edge technology can significantly expand the range of experiments that can be performed. Recently, the CRISPR-Cas13d system has been successfully adapted for use in zebrafish, medaka, killifish, and mouse embryos to achieve targeted gene expression knockdown. Despite its success in other animal models, no prior study has explored the potential of CRISPR-Cas13d in the chick. Here, we present an adaptation of the CRISPR-Cas13d system to achieve targeted gene expression knockdown in the chick embryo. As proof-of-principle, we demonstrate the knockdown of PAX7, an early neural crest marker. Application of this adapted CRISPR-Cas13d technique resulted in effective knockdown of PAX7 expression and function, comparable to knockdown achieved by translation-blocking morpholino. CRISPR-Cas13d complements preexisting knockdown tools such as CRISPR-Cas9 and morpholinos, thereby expanding the experimental potential and versatility of the chick model system.
]]></description>
<dc:creator>Kim, M.</dc:creator>
<dc:creator>Hutchins, E. J.</dc:creator>
<dc:date>2024-08-04</dc:date>
<dc:identifier>doi:10.1101/2024.08.03.606488</dc:identifier>
<dc:title><![CDATA[CRISPR-Cas13d as a molecular tool to achieve targeted gene expression knockdown in chick embryos]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.13.607777v1?rss=1">
<title>
<![CDATA[
Torsional Twist of the SARS-CoV and SARS-CoV-2 SUD-N and SUD-M domains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.13.607777v1?rss=1</link>
<description><![CDATA[
Coronavirus non-structural protein 3 (nsp3) forms hexameric crowns of pores in the double membrane vacuole that houses the replication-transcription complex. Nsp3 in SARS-like viruses has three unique domains absent in other coronavirus nsp3 proteins. Two of these, SUD-N (Macrodomain 2) and SUD-M (Macrodomain 3), form two lobes connected by a peptide linker and an interdomain disulfide bridge. We resolve the first complete x-ray structure of SARS-CoV SUD-N/M as well as a mutant variant of SARS-CoV-2 SUD-N/M modified to restore cysteines for interdomain disulfide bond naturally lost by evolution. Comparative analysis of all structures revealed SUD-N and SUD-M are not rigidly associated, but rather, have significant rotational flexibility. Phylogenetic analysis supports that the disulfide bond cysteines are also absent in pangolin-SARS and closely related viruses, consistent with pangolins being the presumed intermediate host in the emergence of SARS-CoV-2. The absence of these cysteines does not impact viral replication or protein translation.
]]></description>
<dc:creator>Rosas-Lemus, M.</dc:creator>
<dc:creator>Minasov, G.</dc:creator>
<dc:creator>Brunzelle, J. S.</dc:creator>
<dc:creator>Taha, T. Y.</dc:creator>
<dc:creator>Lemak, S.</dc:creator>
<dc:creator>Yin, S.</dc:creator>
<dc:creator>Shuvalova, L.</dc:creator>
<dc:creator>Rosecrans, J.</dc:creator>
<dc:creator>Khanna, K.</dc:creator>
<dc:creator>Seifert, H. S.</dc:creator>
<dc:creator>Savchenko, A.</dc:creator>
<dc:creator>Stogios, P. J.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Satchell, K. J.</dc:creator>
<dc:date>2024-08-14</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607777</dc:identifier>
<dc:title><![CDATA[Torsional Twist of the SARS-CoV and SARS-CoV-2 SUD-N and SUD-M domains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.13.607810v1?rss=1">
<title>
<![CDATA[
The brain simulates actions and their consequences during REM sleep 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.13.607810v1?rss=1</link>
<description><![CDATA[
Vivid dreams mostly occur during a phase of sleep called REM1-5. During REM sleep, the brains internal representation of direction keeps shifting like that of an awake animal moving through its environment6-8. What causes these shifts, given the immobility of the sleeping animal? Here we show that the superior colliculus of the mouse, a motor command center involved in orienting movements9-15, issues motor commands during REM sleep, e.g. turn left, that are similar to those issued in the awake behaving animal. Strikingly, these motor commands, despite not being executed, shift the internal representation of direction as if the animal had turned. Thus, during REM sleep, the brain simulates actions by issuing motor commands that, while not executed, have consequences as if they had been. This study suggests that the sleeping brain, while disengaged from the external world, uses its internal model of the world to simulate interactions with it.
]]></description>
<dc:creator>Senzai, Y.</dc:creator>
<dc:creator>Scanziani, M.</dc:creator>
<dc:date>2024-08-16</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607810</dc:identifier>
<dc:title><![CDATA[The brain simulates actions and their consequences during REM sleep]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.15.607661v1?rss=1">
<title>
<![CDATA[
Perinatal liver inflammation is associated with persistent elevation of CXCL10 and its canonical receptor CXCR3 on common myeloid progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.15.607661v1?rss=1</link>
<description><![CDATA[
Biliary atresia (BA) is a leading cause of liver failure in infants. Despite effective surgical drainage, patients with BA exhibit attenuated immune responses to childhood vaccines, suggesting there are long-lasting alterations to immune function. The perinatal liver is home to hematopoietic stem and progenitor cells (HSPCs) and serves as the epicenter for rapidly progressive and significantly morbid inflammatory diseases like BA. We have previously established the role of neonatal myeloid progenitors in the pathogenesis of perinatal liver inflammation (PLI) and hypothesize that PLI leads to long-term changes to HSPCs in mice that recovered from PLI. To test this hypothesis, we compared the changes that occur to HSPCs and mature myeloid populations in the bone marrow of adult mice during homeostasis and during PLI. Our results demonstrate that HSPCs from animals that recover from PLI ("PLI-recovered") undergo long-term expansion with a reduced proliferative capacity. Notably, PLI leads to persistent activation of common myeloid progenitors through the involvement of CXCL10 and its canonical receptor, CXCR3. Our data suggests that the CXCR3-CXCL10 axis may mediate the changes in HSPCs that lead to altered immune function observed in BA, providing support for a targetable pathway to mitigate the detrimental long-term immune effects observed in patients with BA.
]]></description>
<dc:creator>Alkhani, A.</dc:creator>
<dc:creator>Baskaran, S.</dc:creator>
<dc:creator>Murti, A.</dc:creator>
<dc:creator>Rapp, B.</dc:creator>
<dc:creator>Levy, C. S.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Nijagal, A.</dc:creator>
<dc:date>2024-08-19</dc:date>
<dc:identifier>doi:10.1101/2024.08.15.607661</dc:identifier>
<dc:title><![CDATA[Perinatal liver inflammation is associated with persistent elevation of CXCL10 and its canonical receptor CXCR3 on common myeloid progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.16.608180v1?rss=1">
<title>
<![CDATA[
Quantized multi-task learning for context-specific representations of gene network dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.16.608180v1?rss=1</link>
<description><![CDATA[
While often represented as static entities, gene networks are highly context-dependent. Here, we developed a multi-task learning strategy to yield context-specific representations of gene network dynamics. We assembled a corpus comprising [~]103 million human single-cell transcriptomes from a broad range of tissues and diseases and performed a two stage pretraining, first with non-malignant cells to generate a foundational model and then with continual learning on cancer cells to tune the model to the cancer domain. We performed multi-task learning with the foundational model to learn context-specific representations of a broad range of cell types, tissues, developmental stages, and diseases. We then leveraged the cancer-tuned model to jointly learn cell states and predict tumor-restricting factors within the colorectal tumor microenvironment. Model quantization allowed resource-efficient fine-tuning and inference while preserving biological knowledge. Overall, multi-task learning enables context-specific disease modeling that can yield contextual predictions of candidate therapeutic targets for human disease.
]]></description>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Venkatesh, M. S.</dc:creator>
<dc:creator>Gomez Ortega, J.</dc:creator>
<dc:creator>Mahesh, S. V.</dc:creator>
<dc:creator>Nandi, T. N.</dc:creator>
<dc:creator>Madduri, R. K.</dc:creator>
<dc:creator>Pelka, K.</dc:creator>
<dc:creator>Theodoris, C. V.</dc:creator>
<dc:date>2024-08-19</dc:date>
<dc:identifier>doi:10.1101/2024.08.16.608180</dc:identifier>
<dc:title><![CDATA[Quantized multi-task learning for context-specific representations of gene network dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.16.608251v1?rss=1">
<title>
<![CDATA[
A virtual clinical trial of psychedelics to treat patients with disorders of consciousness 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.16.608251v1?rss=1</link>
<description><![CDATA[
Disorders of consciousness (DoC), including the unresponsive wakefulness syndrome (UWS) and the minimally conscious state (MCS), have limited treatment options. Recent research suggests that psychedelic drugs, known for their complexity-enhancing properties, could be promising treatments for DoC. This study uses whole-brain computational models to explore this potential. We created individualised models for DoC patients, optimised with empirical fMRI and diffusion-weighted imaging (DWI) data, and simulated the administration of LSD and psilocybin. We used an in-silico perturbation protocol to distinguish between different states of consciousness, including DoC, anaesthesia, and the psychedelic state, and assess the dynamical stability of the brains of DoC patients pre- and post-psychedelic simulation. Our findings indicate that LSD and psilocybin shift DoC patients brains closer to criticality, with a greater effect in MCS patients. In UWS patients, the treatment response correlates with structural connectivity, while in MCS patients, it aligns with baseline functional connectivity. This virtual clinical trial lays a computational foundation for using psychedelics in DoC treatment and highlights the future role of computational modelling in drug discovery and personalised medicine.
]]></description>
<dc:creator>Alnagger, N. L. N.</dc:creator>
<dc:creator>Cardone, P.</dc:creator>
<dc:creator>Martial, C.</dc:creator>
<dc:creator>Sanz Perl, Y.</dc:creator>
<dc:creator>Mindlin, I.</dc:creator>
<dc:creator>Sitt, J. D.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Nutt, D.</dc:creator>
<dc:creator>Mallaroni, P.</dc:creator>
<dc:creator>Mason, N. L.</dc:creator>
<dc:creator>Ramaekers, J. G.</dc:creator>
<dc:creator>Bonhomme, V.</dc:creator>
<dc:creator>Laureys, S.</dc:creator>
<dc:creator>Deco, G.</dc:creator>
<dc:creator>Gosseries, O.</dc:creator>
<dc:creator>Nunez, P.</dc:creator>
<dc:creator>Annen, J.</dc:creator>
<dc:date>2024-08-19</dc:date>
<dc:identifier>doi:10.1101/2024.08.16.608251</dc:identifier>
<dc:title><![CDATA[A virtual clinical trial of psychedelics to treat patients with disorders of consciousness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.21.608784v1?rss=1">
<title>
<![CDATA[
Exploring the diversity of anti-defense systems across prokaryotes, phages, and mobile genetic elements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.21.608784v1?rss=1</link>
<description><![CDATA[
The co-evolution of prokaryotes, phages, and mobile genetic elements (MGEs) over the past billions of years has driven the emergence and diversification of defense and anti-defense systems alike. Anti-defense proteins have diverse functional domains, sequences, and are typically small, creating a challenge to detect anti-defense homologs across the prokaryotic genomes. To date, no tools comprehensively annotate anti-defense proteins within a desired genome or MGE. Here, we developed "AntiDefenseFinder" - a free open-source tool and web service that detects 156 anti-defense systems (of one or more proteins) in any genomic sequence. Using this dataset, we identified 47,981 anti-defense systems distributed across prokaryotes, phage, and MGEs. We found that some genes co-localize in "anti-defense islands", including E. coli T4 and Lambda phages, although many are standalone. Out of the 112 systems detected in bacteria, 100 systems localize only or preferentially in prophages, plasmids, phage satellites, integrons, and integrative and conjugative elements. However, over 80% of anti-Pycsar protein 1 (Apyc1) resides in non-mobile regions of bacteria. Evolutionary and functional analyses revealed that Apyc1 likely originated in bacteria to regulate cNMP signaling, but was co-opted multiple times by phages to overcome cNMP-utilizing defenses. With the AntiDefenseFinder tool, we hope to facilitate the identification of the full repertoire of anti-defense systems in MGEs, the discovery of new protein functions, and a deeper understanding of host-pathogen arms race.
]]></description>
<dc:creator>Tesson, F.</dc:creator>
<dc:creator>Huiting, E.</dc:creator>
<dc:creator>Wei, L.</dc:creator>
<dc:creator>Ren, J.</dc:creator>
<dc:creator>Johnson, M. C.</dc:creator>
<dc:creator>Planel, R.</dc:creator>
<dc:creator>Cury, J.</dc:creator>
<dc:creator>Feng, Y.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Bernheim, A.</dc:creator>
<dc:date>2024-08-21</dc:date>
<dc:identifier>doi:10.1101/2024.08.21.608784</dc:identifier>
<dc:title><![CDATA[Exploring the diversity of anti-defense systems across prokaryotes, phages, and mobile genetic elements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.21.609012v1?rss=1">
<title>
<![CDATA[
Context-specific inhibition of mitochondrial ribosomes by phenicol and oxazolidinone antibiotics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.21.609012v1?rss=1</link>
<description><![CDATA[
The antibiotics chloramphenicol (CHL) and oxazolidinones including linezolid (LZD) are known to inhibit mitochondrial translation. This can result in serious, potentially deadly, side effects when used therapeutically. Although the mechanism by which CHL and LZD inhibit bacterial ribosomes has been elucidated in detail, their mechanism of action against mitochondrial ribosomes has yet to be explored. CHL and oxazolidinones bind to the ribosomal peptidyl transfer center (PTC) of the bacterial ribosome and prevent incorporation of incoming amino acids under specific sequence contexts, causing ribosomes to stall only at certain sequences. Through mitoribosome profiling, we show that inhibition of mitochondrial ribosomes is similarly context-specific - CHL and LZD lead to mitoribosome stalling primarily when there is an alanine, serine, or threonine in the penultimate position of the nascent peptide chain. We further validate context-specific stalling through in vitro translation assays. A high resolution cryo-EM structure of LZD bound to the PTC of the human mitoribosome shows extensive similarity to the mode of bacterial inhibition and also suggests potential avenues for altering selectivity. Our findings could help inform the rational development of future, less mitotoxic, antibiotics, which are critically needed in the current era of increasing antimicrobial resistance.
]]></description>
<dc:creator>Bibel, B.</dc:creator>
<dc:creator>Raskar, T.</dc:creator>
<dc:creator>Couvillion, M.</dc:creator>
<dc:creator>Lee, M.</dc:creator>
<dc:creator>Kleinman, J. I.</dc:creator>
<dc:creator>Takeuchi-Tomita, N.</dc:creator>
<dc:creator>Churchman, L. S.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Fujimori, D. G.</dc:creator>
<dc:date>2024-08-21</dc:date>
<dc:identifier>doi:10.1101/2024.08.21.609012</dc:identifier>
<dc:title><![CDATA[Context-specific inhibition of mitochondrial ribosomes by phenicol and oxazolidinone antibiotics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.22.609145v1?rss=1">
<title>
<![CDATA[
Sensitive and modular amplicon sequencing of Plasmodium falciparum diversity and resistance for research and public health 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.22.609145v1?rss=1</link>
<description><![CDATA[
BackgroundTargeted amplicon sequencing is a powerful and efficient tool for interrogating the Plasmodium falciparum genome, generating actionable data from infections to complement traditional malaria epidemiology. For maximum impact, genomic tools should be multi-purpose, robust, sensitive, and reproducible.

MethodsWe developed, characterized, and implemented MAD4HatTeR, an amplicon sequencing panel based on Multiplex Amplicons for Drug, Diagnostic, Diversity, and Differentiation Haplotypes using Targeted Resequencing, along with a bioinformatic pipeline for data analysis. Additionally, we introduce an analytical approach to detect gene duplications and deletions from amplicon sequencing data. Laboratory control and field samples were used to demonstrate the panels high sensitivity and robustness.

ResultsMAD4HatTeR targets 165 highly diverse loci, focusing on multiallelic microhaplotypes, key markers for drug and diagnostic resistance (including duplications and deletions), and csp and potential vaccine targets. The panel can also detect non-falciparum Plasmodium species. MAD4HatTeR successfully generated data from low-parasite-density dried blood spot and mosquito midgut samples, and detected minor alleles at within-sample allele frequencies as low as 1% with high specificity in high-parasite-density dried blood spot samples. Gene deletions and duplications were reliably detected in mono- and polyclonal controls. Data generated by MAD4HatTeR were highly reproducible across multiple laboratories.

ConclusionsThe successful implementation of MAD4HatTeR in five laboratories, including three in malaria-endemic African countries, showcases its feasibility and reproducibility in diverse settings. MAD4HatTeR is thus a powerful tool for research and a robust resource for malaria public health surveillance and control.
]]></description>
<dc:creator>Aranda-Diaz, A.</dc:creator>
<dc:creator>Vickers, E. N.</dc:creator>
<dc:creator>Murie, K.</dc:creator>
<dc:creator>Palmer, B.</dc:creator>
<dc:creator>Hathaway, N.</dc:creator>
<dc:creator>Gerlovina, I.</dc:creator>
<dc:creator>Boene, S.</dc:creator>
<dc:creator>Garcia-Ulloa, M.</dc:creator>
<dc:creator>Cistero, P.</dc:creator>
<dc:creator>Katairo, T.</dc:creator>
<dc:creator>Semakuba, F. D.</dc:creator>
<dc:creator>Nsengimaana, B.</dc:creator>
<dc:creator>Gwarinda, H.</dc:creator>
<dc:creator>Garcia-Fernandez, C.</dc:creator>
<dc:creator>Da Silva, C.</dc:creator>
<dc:creator>Datta, D.</dc:creator>
<dc:creator>Kiyaga, S.</dc:creator>
<dc:creator>Wiringilimaana, I.</dc:creator>
<dc:creator>Fekele, S. M.</dc:creator>
<dc:creator>Parr, J. B.</dc:creator>
<dc:creator>Conrad, M.</dc:creator>
<dc:creator>Raman, J.</dc:creator>
<dc:creator>Tukwasibwe, S.</dc:creator>
<dc:creator>Ssewanyana, I.</dc:creator>
<dc:creator>Rovira-Vallbona, E.</dc:creator>
<dc:creator>Tato, C. M.</dc:creator>
<dc:creator>Briggs, J.</dc:creator>
<dc:creator>Mayor, A.</dc:creator>
<dc:creator>Greenhouse, B.</dc:creator>
<dc:date>2024-08-22</dc:date>
<dc:identifier>doi:10.1101/2024.08.22.609145</dc:identifier>
<dc:title><![CDATA[Sensitive and modular amplicon sequencing of Plasmodium falciparum diversity and resistance for research and public health]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.23.609244v1?rss=1">
<title>
<![CDATA[
HMGB1 restores a dynamic chromatin environment in the presence of linker histone by deforming nucleosomal DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.23.609244v1?rss=1</link>
<description><![CDATA[
The essential architectural protein HMGB1 increases accessibility of nucleosomal DNA and counteracts the effects of linker histone H1. However, HMGB1 is less abundant than H1 and binds nucleosomes more weakly raising the question of how HMGB1 effectively competes with H1. Here, we show that HMGB1 rescues H1s inhibition of nucleosomal DNA accessibility without displacing H1. HMGB1 also increases the dynamics of condensed, H1-bound chromatin. Cryo-EM shows that HMGB1 binds at internal locations on a nucleosome and locally distorts the DNA. These sites, which are away from the binding site of H1, explain how HMGB1 and H1 co-occupy a nucleosome. Our findings lead to a model where HMGB1 counteracts the activity of H1 by distorting nucleosomal DNA and by contacting the H1 C-terminal tail. Compared to direct competition, nucleosome co-occupancy by HMGB1 and H1 allows a greater diversity of dynamic chromatin states and may be generalizable to other chromatin regulators.
]]></description>
<dc:creator>Saunders, H. S.</dc:creator>
<dc:creator>Chio, U. S.</dc:creator>
<dc:creator>Moore, C. M.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:date>2024-08-24</dc:date>
<dc:identifier>doi:10.1101/2024.08.23.609244</dc:identifier>
<dc:title><![CDATA[HMGB1 restores a dynamic chromatin environment in the presence of linker histone by deforming nucleosomal DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.25.609621v1?rss=1">
<title>
<![CDATA[
Extensive exploration of structure activity relationships for the SARS-CoV-2 macrodomain from shape-based fragment merging and active learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.25.609621v1?rss=1</link>
<description><![CDATA[
The macrodomain contained in the SARS-CoV-2 non-structural protein 3 (NSP3) is required for viral pathogenesis and lethality. Inhibitors that block the macrodomain could be a new therapeutic strategy for viral suppression. We previously performed a large-scale X-ray crystallography-based fragment screen and discovered a sub-micromolar inhibitor by fragment linking. However, this carboxylic acid-containing lead had poor membrane permeability and other liabilities that made optimization difficult. Here, we developed a shape- based virtual screening pipeline - FrankenROCS - to identify new macrodomain inhibitors using fragment X-ray crystal structures. We used FrankenROCS to exhaustively screen the Enamine high-throughput screening (HTS) collection of 2.1 million compounds and selected 39 compounds for testing, with the most potent compound having an IC50 value equal to 130 M. We then paired FrankenROCS with an active learning algorithm (Thompson sampling) to efficiently search the Enamine REAL database of 22 billion molecules, testing 32 compounds with the most potent having an IC50 equal to 220 M. Further optimization led to analogs with IC50 values better than 10 M, with X-ray crystal structures revealing diverse binding modes despite conserved chemical features. These analogs represent a new lead series with improved membrane permeability that is poised for optimization. In addition, the collection of 137 X-ray crystal structures with associated binding data will serve as a resource for the development of structure-based drug discovery methods. FrankenROCS may be a scalable method for fragment linking to exploit ever-growing synthesis-on- demand libraries.
]]></description>
<dc:creator>Correy, G. J.</dc:creator>
<dc:creator>Rachman, M. M.</dc:creator>
<dc:creator>Togo, T.</dc:creator>
<dc:creator>Gahbauer, S.</dc:creator>
<dc:creator>Doruk, Y. U.</dc:creator>
<dc:creator>Stevens, M. G. V.</dc:creator>
<dc:creator>Jaishankar, P.</dc:creator>
<dc:creator>Kelley, B.</dc:creator>
<dc:creator>Goldman, B.</dc:creator>
<dc:creator>Schmidt, M.</dc:creator>
<dc:creator>Kramer, T.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Riley, P.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Renslo, A. R.</dc:creator>
<dc:creator>Walters, W. P.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2024-08-26</dc:date>
<dc:identifier>doi:10.1101/2024.08.25.609621</dc:identifier>
<dc:title><![CDATA[Extensive exploration of structure activity relationships for the SARS-CoV-2 macrodomain from shape-based fragment merging and active learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.27.609995v1?rss=1">
<title>
<![CDATA[
Sustained fertility from first-wave follicle oocytes that pause their growth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.27.609995v1?rss=1</link>
<description><![CDATA[
Ovulation results from the cyclical recruitment of non-renewing, quiescent oocytes for growth. Therefore, the primordial follicles that are established during development from an oocyte encapsulated by granulosa cells are thought to comprise the lifelong ovarian reserve 1-4. However, using oocyte lineage tracing in mice, we observed that a subset of oocytes recruited for growth in the first juvenile wave remain paused for many months before continuing growth, ovulation, fertilization and development into healthy offspring. This small subset of genetically-labeled fetal oocytes, labeled with Sycp3-CreERT2, is distinguished by earlier entry and slower dynamics of meiotic prophase I. While labeled oocytes were initially found in both primordial follicles and growing follicles of the first wave, they disappeared from primordial follicles by puberty. Unexpectedly, these first-wave labeled growing oocytes persisted throughout reproductive lifespan and contributed to offspring at a steady rate beyond 12 months of age, suggesting that follicles can pause mid-growth for extended periods then successfully resume. These results challenge the conclusion from lineage tracing of granulosa cells that first-wave follicles make a limited contribution to fertility5 and furthermore suggest that growth-paused oocytes comprise a second and previously unrecognized ovarian reserve.
]]></description>
<dc:creator>Soygur, B.</dc:creator>
<dc:creator>Gaylord, E. A.</dc:creator>
<dc:creator>Foecke, M. H.</dc:creator>
<dc:creator>Cincotta, S. A.</dc:creator>
<dc:creator>Horan, T. J.</dc:creator>
<dc:creator>Wood, A.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:date>2024-08-28</dc:date>
<dc:identifier>doi:10.1101/2024.08.27.609995</dc:identifier>
<dc:title><![CDATA[Sustained fertility from first-wave follicle oocytes that pause their growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.28.609802v1?rss=1">
<title>
<![CDATA[
Mutant p53 Exploits Enhancers to Elevate Immunosuppressive Chemokine Expression and Impair Immune Checkpoint Inhibitors in Pancreatic Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.28.609802v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer without effective treatments. It is characterized by activating KRAS mutations and p53 alterations. However, how these mutations dysregulate cancer-cell-intrinsic gene programs to influence the immune landscape of the tumor microenvironment (TME) remains poorly understood. Here, we show that p53R172H establishes an immunosuppressive TME, diminishes the efficacy of immune checkpoint inhibitors (ICIs), and enhances tumor growth. Our findings reveal that the upregulation of the immunosuppressive chemokine Cxcl1 mediates these pro-tumorigenic functions of p53R172H. Mechanistically, we show that p53R172H associates with the distal enhancers of the Cxcl1 gene, increasing enhancer activity and Cxcl1 expression. p53R172H occupies these enhancers in an NF-{kappa}B-pathway-dependent manner, suggesting NF-{kappa}Bs role in recruiting p53R172H to the Cxcl1 enhancers. Our work uncovers how a common mutation in a tumor-suppressor transcription factor appropriates enhancers, stimulating chemokine expression and establishing an immunosuppressive TME that diminishes ICI efficacy in PDAC.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=100 SRC="FIGDIR/small/609802v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Mahat, D. B.</dc:creator>
<dc:creator>Kumra, H.</dc:creator>
<dc:creator>Castro, S. A.</dc:creator>
<dc:creator>Metcalf, E.</dc:creator>
<dc:creator>Nguyen, K.</dc:creator>
<dc:creator>Morisue, R.</dc:creator>
<dc:creator>Ho, W. W.</dc:creator>
<dc:creator>Chen, I.</dc:creator>
<dc:creator>Sullivan, B.</dc:creator>
<dc:creator>Yim, L. K.</dc:creator>
<dc:creator>Singh, A.</dc:creator>
<dc:creator>Fu, J.</dc:creator>
<dc:creator>Waterton, S. K.</dc:creator>
<dc:creator>Cheng, Y.-C.</dc:creator>
<dc:creator>Roberge, S.</dc:creator>
<dc:creator>Moiso, E.</dc:creator>
<dc:creator>Chauhan, V. P.</dc:creator>
<dc:creator>Moura Silva, H.</dc:creator>
<dc:creator>Spranger, S.</dc:creator>
<dc:creator>Jain, R. K.</dc:creator>
<dc:creator>Sharp, P. A.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.28.609802</dc:identifier>
<dc:title><![CDATA[Mutant p53 Exploits Enhancers to Elevate Immunosuppressive Chemokine Expression and Impair Immune Checkpoint Inhibitors in Pancreatic Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.29.610377v1?rss=1">
<title>
<![CDATA[
Coordinated Tbx3 / Tbx5 transcriptional control of the adult ventricular conduction system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.29.610377v1?rss=1</link>
<description><![CDATA[
The cardiac conduction system (CCS) orchestrates the electrical impulses that enable coordinated contraction of the cardiac chambers. The T-box transcription factors TBX3 and TBX5 are required for cardiac conduction system development and associated with overlapping and distinct human cardiac conduction system diseases. We evaluated the coordinated role of Tbx3 and Tbx5 in the murine ventricular conduction system (VCS). We engineered a compound Tbx3:Tbx5 conditional knockout allele for both genes located in cis on mouse chromosome 5. Conditional deletion of both T-box transcriptional factors in the ventricular conduction system, using the VCS-specific MinK:Cre, caused loss of VCS function and molecular identity. Combined Tbx3 and Tbx5 deficiency in the adult VCS led to conduction defects, including prolonged PR and QRS intervals and elevated susceptibility to ventricular tachycardia. These electrophysiological defects occurred prior to detectable alterations in cardiac contractility or histologic morphology, indicative of a primary conduction system defect. Tbx3:Tbx5 double knockout VCS cardiomyocytes revealed a transcriptional shift towards non-CCS-specialized working myocardium, indicating a change to their cellular identity. Furthermore, optical mapping revealed a loss of VCS-specific conduction system propagation. Collectively, these findings indicate that Tbx3 and Tbx5 coordinate to control VCS molecular fate and function, with implications for understanding cardiac conduction disorders in humans.
]]></description>
<dc:creator>Burnicka-Turek, O.</dc:creator>
<dc:creator>Trampel, K. A.</dc:creator>
<dc:creator>Laforest, B.</dc:creator>
<dc:creator>Broman, M. T.</dc:creator>
<dc:creator>Khan, Z.</dc:creator>
<dc:creator>Rytkin, E.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Schaffer, E.</dc:creator>
<dc:creator>Gadek, M.</dc:creator>
<dc:creator>Shen, K. M.</dc:creator>
<dc:creator>Efimov, I. R.</dc:creator>
<dc:creator>Moskowitz, I. P.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.29.610377</dc:identifier>
<dc:title><![CDATA[Coordinated Tbx3 / Tbx5 transcriptional control of the adult ventricular conduction system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.29.610420v1?rss=1">
<title>
<![CDATA[
Comprehensive profiling of migratory primordial germ cells reveals niche-specific differences in non-canonical Wnt and Nodal-Lefty signaling in anterior vs posterior migrants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.29.610420v1?rss=1</link>
<description><![CDATA[
Mammalian primordial germ cells (PGCs) migrate asynchronously through the embryonic hindgut and dorsal mesentery to reach the gonads. We previously found that interaction with different somatic niches regulates PGC proliferation along the migration route. To characterize transcriptional heterogeneity of migrating PGCs and their niches, we performed single-cell RNA sequencing of 13,262 mouse PGCs and 7,868 surrounding somatic cells during migration (E9.5, E10.5, E11.5) and in anterior versus posterior locations to enrich for leading and lagging migrants. Analysis of PGCs by position revealed dynamic gene expression changes between faster or earlier migrants in the anterior and slower or later migrants in the posterior at E9.5; these differences include migration-associated actin polymerization machinery and epigenetic reprogramming-associated genes. We furthermore identified changes in signaling with various somatic niches, notably strengthened interactions with hindgut epithelium via non-canonical WNT (ncWNT) in posterior PGCs compared to anterior. Reanalysis of a previously published dataset suggests that ncWNT signaling from the hindgut epithelium to early migratory PGCs is conserved in humans. Trajectory inference methods identified putative differentiation trajectories linking cell states across timepoints and from posterior to anterior in our mouse dataset. At E9.5, we mainly observed differences in cell adhesion and actin cytoskeletal dynamics between E9.5 posterior and anterior migrants. At E10.5, we observed divergent gene expression patterns between putative differentiation trajectories from posterior to anterior including Nodal signaling response genes Lefty1, Lefty2, and Pycr2 and reprogramming factors Dnmt1, Prc1, and Tet1. At E10.5, we experimentally validated anterior migrant-specific Lefty1/2 upregulation via whole-mount immunofluorescence staining for LEFTY1/2 and phosphorylated SMAD2/3, suggesting that elevated autocrine Nodal signaling in migrating PGCs occurs as they near the gonadal ridges. Together, this positional and temporal atlas of mouse PGCs supports the idea that niche interactions along the migratory route elicit changes in proliferation, actin dynamics, pluripotency, and epigenetic reprogramming.
]]></description>
<dc:creator>Jaszczak, R. G.</dc:creator>
<dc:creator>Zussman, J. W.</dc:creator>
<dc:creator>Wagner, D. E.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.29.610420</dc:identifier>
<dc:title><![CDATA[Comprehensive profiling of migratory primordial germ cells reveals niche-specific differences in non-canonical Wnt and Nodal-Lefty signaling in anterior vs posterior migrants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.30.610521v1?rss=1">
<title>
<![CDATA[
A Novel Approach to Reducing Spinal Implant-Associated Infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.30.610521v1?rss=1</link>
<description><![CDATA[
Implant-associated infection is a severe complication of instrumented spinal surgery. Preventive measures against biofilm formation, such as chemical modifications of devices with antibiotics or bactericidal substances, have been explored to decrease infection rates and enhance therapeutic outcomes. However, this approach remains challenging, and the safety issues associated with antibiotic resistance and cellular toxicity need to be addressed. Hence, there is an unmet clinical need for implants that resist biofilm formation. Ultrafine-grained material (UFG), which has a nanoscale grain structure, is a novel form of metal with exceptional mechanical and biocompatibility properties that may also have antimicrobial activity. This study investigates the extent to which UFG stainless steel implants resist biofilm formation by Staphylococcus aureus in vitro and in vivo. The results show that the UFG stainless steel wire significantly reduced biofilm formation at early time points after bacterial infection compared to a standard wire. In mouse models of implant-associated infection below the skin or the fascia, the UFG wire had significantly less biofilm formation at early time points. The UFG wire appeared to delay initial bacterial adhesion and biofilm maturation. These findings suggest that UFG implants may resist initial biofilm formation and maturation, leading to promising antibacterial materials for patients.
]]></description>
<dc:creator>Nishizawa, M.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Serhan, H.</dc:creator>
<dc:creator>Saleh, B.</dc:creator>
<dc:creator>Marcucio, R. S.</dc:creator>
<dc:creator>Morioka, K.</dc:creator>
<dc:date>2024-08-31</dc:date>
<dc:identifier>doi:10.1101/2024.08.30.610521</dc:identifier>
<dc:title><![CDATA[A Novel Approach to Reducing Spinal Implant-Associated Infections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.30.610436v1?rss=1">
<title>
<![CDATA[
Deciphering regulation of FOXP3 expression in human conventional T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.30.610436v1?rss=1</link>
<description><![CDATA[
FOXP3 is a lineage-defining transcription factor (TF) for immune-suppressive regulatory T cells (Tregs). While mice exclusively express FOXP3 in Tregs, humans also transiently express FOXP3 in stimulated conventional CD4+ T cells (Tconvs). Mechanisms governing these distinct expression patterns remain unknown. Here, we performed CRISPR screens tiling the FOXP3 locus and targeting TFs in human Tregs and Tconvs to discover cis-regulatory elements (CREs) and trans-regulators of FOXP3. Tconv FOXP3 expression depended on a subset of Treg CREs and Tconv-selective positive (TcNS+) and negative (TcNS-) CREs. The CREs are occupied and regulated by TFs we identified as critical regulators of FOXP3. Finally, mutagenesis of murine TcNS- revealed that it is critical for restriction of FOXP3 expression to Tregs. We discover CRE and TF circuitry controlling FOXP3 expression and reveal evolution of mechanisms regulating a gene indispensable to immune homeostasis.

HighlightsO_LIComprehensive CRISPR maps of CREs and TFs controlling FOXP3 in human Tregs and Tconvs
C_LIO_LIKey TFs that control FOXP3 directly occupy and regulate CREs forming TF-CRE circuits
C_LIO_LIA previously unknown negative CRE stringently restricts FOXP3 to Tregs in mice
C_LI
]]></description>
<dc:creator>Umhoefer, J. M.</dc:creator>
<dc:creator>Arce, M. M.</dc:creator>
<dc:creator>Dajani, R.</dc:creator>
<dc:creator>Belk, J. A.</dc:creator>
<dc:creator>Mowery, C. T.</dc:creator>
<dc:creator>Nguyen, V.</dc:creator>
<dc:creator>Gowen, B. G.</dc:creator>
<dc:creator>Simeonov, D. R.</dc:creator>
<dc:creator>Curie, G. L.</dc:creator>
<dc:creator>Corn, J. E.</dc:creator>
<dc:creator>Chang, H. Y.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2024-09-02</dc:date>
<dc:identifier>doi:10.1101/2024.08.30.610436</dc:identifier>
<dc:title><![CDATA[Deciphering regulation of FOXP3 expression in human conventional T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.02.610853v1?rss=1">
<title>
<![CDATA[
Methods for high throughput discovery of fluoroprobes that recognize tau fibril polymorphs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.02.610853v1?rss=1</link>
<description><![CDATA[
Aggregation of microtubule-associated protein tau (MAPT/tau) into conformationally distinct fibrils underpins neurodegenerative tauopathies. Fluorescent probes (fluoroprobes), such as thioflavin T (ThT), have been essential tools for studying tau aggregation; however, most of them do not discriminate between amyloid fibril conformations (polymorphs). This gap is due, in part, to a lack of high-throughput methods for screening large, diverse chemical collections. Here, we leverage advances in protein adaptive differential scanning fluorimetry (paDSF) to screen the Aurora collection of 300+ fluorescent dyes against multiple synthetic tau fibril polymorphs. This screen, coupled with orthogonal secondary assays, revealed pan-fibril binding chemotypes, as well as fluoroprobes selective for subsets of fibrils. One fluoroprobe recognized tau pathology in ex vivo brain slices from Alzheimers disease patients. We propose that these scaffolds represent entry points for development of selective fibril ligands and, more broadly, that high throughput, fluorescence-based dye screening is a platform for their discovery.
]]></description>
<dc:creator>Carroll, E. C.</dc:creator>
<dc:creator>Yang, H. E.</dc:creator>
<dc:creator>Jones, J. G.</dc:creator>
<dc:creator>Oehler, A.</dc:creator>
<dc:creator>Charvat, A. F.</dc:creator>
<dc:creator>Montgomery, K. M.</dc:creator>
<dc:creator>Yung, A.</dc:creator>
<dc:creator>Millbern, Z.</dc:creator>
<dc:creator>Vinueza, N. R.</dc:creator>
<dc:creator>Degrado, W. F.</dc:creator>
<dc:creator>Mordes, D. A.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:date>2024-09-02</dc:date>
<dc:identifier>doi:10.1101/2024.09.02.610853</dc:identifier>
<dc:title><![CDATA[Methods for high throughput discovery of fluoroprobes that recognize tau fibril polymorphs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.02.610895v1?rss=1">
<title>
<![CDATA[
The miR-290 and miR-302 clusters are essential for reprogramming of fibroblasts to induced pluripotent stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.02.610895v1?rss=1</link>
<description><![CDATA[
The miR-290 and miR-302 clusters of microRNAs are highly expressed in naive and primed pluripotent stem cells, respectively. Ectopic expression of the embryonic stem cell-specific cell cycle regulating (ESCC) family of microRNAs arising from these two clusters dramatically enhances the reprogramming of both mouse and human somatic cells to induced pluripotency. Here, we used genetic knockouts to dissect the requirement for the miR-290 and miR-302 clusters during the reprogramming of mouse fibroblasts into induced pluripotent stem cells (iPSCs) with retrovirally introduced Oct4, Sox2, and Klf4. Knockout of either cluster alone did not negatively impact the efficiency of reprogramming. Resulting cells appeared identical to their embryonic stem cell microRNA cluster knockout counterparts. In contrast, the combined loss of both clusters blocked the formation of iPSCs. While rare double knockout clones could be isolated, they showed a dramatically reduced proliferation rate, a persistent inability to fully silence the exogenously introduced pluripotency factors, and a transcriptome distinct from individual miR-290 or miR-302 mutant ESC and iPSCs. Taken together, our data show that miR-290 and miR-302 are essential yet interchangeable in reprogramming to the induced pluripotent state.

Impact StatementThe process by which somatic cell reprogramming yields induced pluripotent stem cells (iPSCs) is incompletely understood. MicroRNAs from the miR-290 and miR-302 clusters have been shown to greatly increase reprogramming efficiency, but their requirement in the process has not been studied. Here, we examine this requirement by genetically removing the miRNA clusters in somatic cells. We discover that somatic cells lacking either, but not both, of these miRNA clusters can form iPSC cells. This work thus provides new important insight into mechanisms underlying reprogramming to pluripotency.
]]></description>
<dc:creator>Ye, J.</dc:creator>
<dc:creator>Boileau, R. M.</dc:creator>
<dc:creator>Parchem, R. J.</dc:creator>
<dc:creator>Judson-Torres, R. L.</dc:creator>
<dc:creator>Blelloch, R.</dc:creator>
<dc:date>2024-09-03</dc:date>
<dc:identifier>doi:10.1101/2024.09.02.610895</dc:identifier>
<dc:title><![CDATA[The miR-290 and miR-302 clusters are essential for reprogramming of fibroblasts to induced pluripotent stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.04.611325v1?rss=1">
<title>
<![CDATA[
Structure and inhibition mechanisms of Mycobacterium tuberculosis essential transporterefflux protein A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.04.611325v1?rss=1</link>
<description><![CDATA[
A broad chemical genetics screen in Mycobacterium tuberculosis (Mtb) to identify inhibitors of established or previously untapped targets for therapeutic development yielded compounds (BRD-8000.3 and BRD-9327) that inhibit the essential efflux pump EfpA. To understand the mechanisms of inhibition by these compounds, we determined the structures of EfpA with inhibitors bound at 2.7 -3.4 [A] resolution. Our structures reveal different mechanisms of inhibition for the two inhibitors. BRD-8000.3 binds in a tunnel making contact with the lipid bilayer and extending toward the central cavity to displace the fatty acid chain of a lipid molecule bound in the apo structure, suggesting its blocking of an access route for a natural lipidic substrate, in contrast to its uncompetitive mechanism for the small molecule substrate ethidium bromide which likely enters through an alternative tunnel. Meanwhile, BRD-9327 binds in the outer vestibule without complete blockade of the substrate path to the outside, suggesting its possible inhibition of the dynamical motion necessary for "alternate access" to the two different sides of the membrane, as is characteristic of major facilitator superfamily (MFS) transporters. Both inhibitors may have a role in inhibiting the "alternate access" mechanism that could account for the uncompetitive nature of their efflux of some substrates. Our results explain the basis of the synergy of these inhibitors and their potential for combination in a multi drug strategy for anti-tuberculosis therapy. They also potentially point to a possible function for this essential efflux pump as a lipid transporter. The structures provide a foundation for rational modification of these inhibitors to increase potency.
]]></description>
<dc:creator>Khandelwal, N. K.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Gomez, J. E.</dc:creator>
<dc:creator>Barkho, S.</dc:creator>
<dc:creator>Guan, Z.</dc:creator>
<dc:creator>Eng, A. Y.</dc:creator>
<dc:creator>Kawate, T.</dc:creator>
<dc:creator>Balasubramani, S. G.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Hung, D. T.</dc:creator>
<dc:creator>Stroud, R. M.</dc:creator>
<dc:date>2024-09-05</dc:date>
<dc:identifier>doi:10.1101/2024.09.04.611325</dc:identifier>
<dc:title><![CDATA[Structure and inhibition mechanisms of Mycobacterium tuberculosis essential transporterefflux protein A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.03.610947v1?rss=1">
<title>
<![CDATA[
Vascular HIF2 signaling prevents cardiomegaly, alveolar congestion and capillary remodeling during chronic hypoxia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.03.610947v1?rss=1</link>
<description><![CDATA[
Hypoxia is associated with the onset of cardiovascular diseases including cardiac hypertrophy and pulmonary arterial hypertension (PAH). Endothelial HIF2 signaling mediates pulmonary arterial remodeling and subsequent right ventricular systolic pressure (RVSP) elevation during chronic hypoxia, encouraging novel therapeutic opportunities for PAH based on specific HIF2 inhibitors. Nevertheless, HIF2 relevance beyond the pulmonary endothelium or in the cardiac adaptation to hypoxia remains elusive. Wilms tumor 1 lineage contributes to heart and lung vascular compartments including pericytes, endothelial and smooth muscle cells. Here we describe the response to chronic hypoxia of a novel HIF2 mutant mouse model in the Wt1 lineage (Hif2/Wt1 cKO). Hif2/Wt1 cKO is protected against pulmonary remodeling and increased RVSP induced by hypoxia, but displays alveolar congestion, inflammation and hemorrhages associated with microvascular instability. Furthermore, lack of HIF2 in the Wt1 lineage leads to cardiomegaly, capillary remodeling, right and left ventricular hypertrophy, systolic dysfunction and left ventricular dilation, suggesting pulmonary-independent cardiac direct roles of HIF2 in hypoxia. These structural defects are partially restored upon reoxygenation, while functional parameters remain altered. Our results suggest that cardiopulmonary HIF2 signaling prevents excessive vascular proliferation during chronic hypoxia and define novel protective roles of HIF2 to warrant stable microvasculature and organ function.
]]></description>
<dc:creator>Albendea-Gomez, T.</dc:creator>
<dc:creator>Mendoza-Tamajon, S.</dc:creator>
<dc:creator>Castro-Mecinas, R.</dc:creator>
<dc:creator>Escobar, B.</dc:creator>
<dc:creator>Rocha, S. F.</dc:creator>
<dc:creator>Urra-Balduz, S.</dc:creator>
<dc:creator>Nicolas-Avila, J. A.</dc:creator>
<dc:creator>Oliver, E.</dc:creator>
<dc:creator>Villalba-Orero, M.</dc:creator>
<dc:creator>Martin-Puig, S.</dc:creator>
<dc:date>2024-09-06</dc:date>
<dc:identifier>doi:10.1101/2024.09.03.610947</dc:identifier>
<dc:title><![CDATA[Vascular HIF2 signaling prevents cardiomegaly, alveolar congestion and capillary remodeling during chronic hypoxia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.04.609712v1?rss=1">
<title>
<![CDATA[
Identifying deleterious noncoding variation through gain and loss of CTCF binding activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.04.609712v1?rss=1</link>
<description><![CDATA[
Noncoding single nucleotide variants are the predominant class of genetic variation in whole genome sequencing and are key drivers of phenotypic variation. However, their functional annotation remains challenging. To address this, we develop a hypothesis-driven functional annotation scheme for CTCF binding sites given CTCFs critical roles in gene regulation and extensive profiling in regulatory datasets. We synthesize CTCFs binding patterns at 1,063,879 genomic loci across 214 biological contexts into a summary metric, which we refer to as binding activity. We find that binding activity is significantly enriched for both conserved nucleotides (Pearson R = 0.31, p < 2.2 x 10-16) and sequences that contain high-quality CTCF binding motifs (Pearson R = 0.63, p = 2.9 x 10-12). We then integrate binding activity with high confidence change in precision weight matrix scores. By applying this framework to 1,253,330 SNVs in gnomAD, we explore signatures of selection acting against the disruption of CTCF binding. We find a strong, positive relationship between the mutability adjusted proportion of singletons (MAPS) metric and the loss of CTCF binding at loci with high in vitro activity (Pearson R = 0.67, p = 1.5 x 10-14). To contextualize these findings, we apply MAPS to other functional classes of variation and find that a subset of 198,149 loss of CTCF binding variants are observed as infrequently as missense variants. This work implicates these thousands of rare, noncoding variants that disrupt CTCF binding for further functional studies while providing a blueprint for the interpretable annotation of noncoding variants.
]]></description>
<dc:creator>Tubbs, C.</dc:creator>
<dc:creator>Benton, M. L.</dc:creator>
<dc:creator>McArthur, E.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:creator>Ruderfer, D. M.</dc:creator>
<dc:date>2024-09-08</dc:date>
<dc:identifier>doi:10.1101/2024.09.04.609712</dc:identifier>
<dc:title><![CDATA[Identifying deleterious noncoding variation through gain and loss of CTCF binding activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.09.611948v1?rss=1">
<title>
<![CDATA[
How a pathogenic mutation impairs Hsp60 functional dynamics from monomeric to fully assembled states 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.09.611948v1?rss=1</link>
<description><![CDATA[
Heat Shock Protein 60 kDa (Hsp60) is a mitochondrial chaperonin that cooperates with Hsp10 to drive the correct folding of client proteins. Monomers M of Hsp60 (featuring equatorial, intermediate, and apical domains) first assemble into 7-meric Single rings (S), then pairs of S interface equatorially to form 14-meric Double rings (D) that accommodate clients into their lumen. Recruitment of 7 Hsp10 molecules per pole turns D into a 28-meric Football-shaped complex (F). Sequential hydrolysis of ATP present in each Hsp60 unit of F finally drives client folding and F disassembly. Equatorial domain mutation V72I occurs in SPG13, a form of hereditary spastic paraplegia: while distal to the active site, this severely impairs the chaperone cycle and stability. To understand the molecular bases of this impairment we have run atomistic molecular dynamics (MD) simulations of M, S, D, and F for both WT and mutant Hsp60, with two catalytically relevant Hsp60 aspartates in D and F modelled in three different protonation states. Additionally, D in one protonation state was modelled post-hydrolysis (total production time: 36 {micro}s). By combining complementary experimental and computational approaches for the analysis of functional dynamics and allosteric mechanisms, we consistently find that mutation V72I significantly rewires allosteric routes present in WT Hsp60 across its complexes, from isolated M units right up to F, rigidifying them--as observed experimentally--by introducing a direct allosteric link between equatorial and apical Hsp60 domains that bypasses the ATP binding site (wherein we observe the alteration of mechanisms driving reactivity). Our results reveal a multiscale complexity of functional mechanisms for Hsp60 and its pathogenic mutant, and may lay the foundation for the design of experiments to fully understand both variants.
]]></description>
<dc:creator>Torielli, L.</dc:creator>
<dc:creator>Guarra, F.</dc:creator>
<dc:creator>Shao, H.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Serapian, S. A.</dc:creator>
<dc:creator>Colombo, G.</dc:creator>
<dc:date>2024-09-09</dc:date>
<dc:identifier>doi:10.1101/2024.09.09.611948</dc:identifier>
<dc:title><![CDATA[How a pathogenic mutation impairs Hsp60 functional dynamics from monomeric to fully assembled states]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.04.610637v1?rss=1">
<title>
<![CDATA[
Ketogenic interventions restore cognition and modulate peripheral metabolic dysfunctions in Alzheimer's disease mouse models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.04.610637v1?rss=1</link>
<description><![CDATA[
Lifestyle factors modulate dementia risk. We investigated mechanisms of dementia risk reduction by emerging dietary ketogenic interventions. We show that distinct interventions, a medium-chain triglycerides (MCT)-enriched diet and a carbohydrate-free, high-fat diet (CFHF), improve cognition and dendritic spine density of memory-associated hippocampal neurons in two mouse models of Alzheimers disease (AD). Only the CFHF diet drove increased circulating ketones, suggesting distinct underlying mechanisms. AD mice exhibited baseline and diet-induced susceptibility to peripheral metabolic disturbances that were improved by MCT and exacerbated by CFHF diets. Prominent AD-associated dysregulation of the liver transcriptome was largely restored by both interventions, but MCT also downregulated lipogenic enzymes and did not trigger a CFHF-like inflammatory signature. Novel AD- and diet-induced plasmatic changes in hormones and lipid species were identified. Thus, different ketogenic interventions yield cognitive benefits in AD models while showing intervention-specific modulation of peripheral metabolic defects, with implications for design of therapeutic ketogenic strategies.
]]></description>
<dc:creator>M'Bra, P. E. H.</dc:creator>
<dc:creator>Hamilton, L. K.</dc:creator>
<dc:creator>Moquin-Beaudry, G.</dc:creator>
<dc:creator>Aubin, M.</dc:creator>
<dc:creator>Lyn-Mangahas, C.</dc:creator>
<dc:creator>Pratesi, F.</dc:creator>
<dc:creator>Brunet, E.</dc:creator>
<dc:creator>Castonguay, A.</dc:creator>
<dc:creator>Mailloux, S.</dc:creator>
<dc:creator>Galoppin, M.</dc:creator>
<dc:creator>Bernier, M.</dc:creator>
<dc:creator>Turri, M.</dc:creator>
<dc:creator>Vachon, A.</dc:creator>
<dc:creator>Mayhue, M.</dc:creator>
<dc:creator>Aumont, A.</dc:creator>
<dc:creator>Tetreault, M.</dc:creator>
<dc:creator>Plourde, M.</dc:creator>
<dc:creator>Fernandes, K. J. L.</dc:creator>
<dc:date>2024-09-10</dc:date>
<dc:identifier>doi:10.1101/2024.09.04.610637</dc:identifier>
<dc:title><![CDATA[Ketogenic interventions restore cognition and modulate peripheral metabolic dysfunctions in Alzheimer's disease mouse models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.10.611504v1?rss=1">
<title>
<![CDATA[
ATP-dependent remodeling of chromatin condensates uncovers distinct mesoscale effects of two remodelers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.10.611504v1?rss=1</link>
<description><![CDATA[
ATP-dependent chromatin remodeling enzymes mobilize nucleosomes, but how such mobilization affects chromatin condensation is unclear. Here, we investigate effects of two major remodelers, ACF and RSC using chromatin condensates and single-molecule footprinting. We find that both remodelers inhibit the formation of condensed chromatin. However, the remodelers have distinct effects on pre-formed chromatin condensates. ACF spaces nucleosomes without de-condensing the chromatin, explaining how ACF maintains nucleosome organization in transcriptionally repressed genomic regions. In contrast, RSC catalyzes ATP-dependent de-condensation of chromatin. Surprisingly, RSC also drives micron-scale movements of entire condensates. These newly uncovered activities of RSC explain its central role in transcriptional activation. The biological importance of remodelers may thus reflect both their effects on nucleosome mobilization and the corresponding consequences on chromatin dynamics at the mesoscale.
]]></description>
<dc:creator>Moore, C. M.</dc:creator>
<dc:creator>Kaur, U.</dc:creator>
<dc:creator>Wong, E.</dc:creator>
<dc:creator>Chio, U. S. M.</dc:creator>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Ostrowski, M.</dc:creator>
<dc:creator>Wu, K.</dc:creator>
<dc:creator>Irkliyenko, I.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:date>2024-09-10</dc:date>
<dc:identifier>doi:10.1101/2024.09.10.611504</dc:identifier>
<dc:title><![CDATA[ATP-dependent remodeling of chromatin condensates uncovers distinct mesoscale effects of two remodelers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.08.611839v1?rss=1">
<title>
<![CDATA[
ElixirSeeker: A Machine Learning Framework Utilizing Attention-Driven Fusion of Molecular Fingerprints for the Discovery of Anti-Aging Compounds 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.08.611839v1?rss=1</link>
<description><![CDATA[
Despite the growing interest in anti-aging drug development, high cost and low success rate pose a significant challenge. We present ElixirSeeker, a new machine-learning framework designed to help speed up the discovery of potential anti-aging compounds by utilizing the attention-driven fusion of molecular fingerprints. Our approach integrates molecular fingerprints generated by different algorithms and utilizes XGBoost to select optimal fingerprint lengths. Subsequently, we assign weights to the molecular fingerprints and employ Kernel Principal Component Analysis (KPCA) to reduce dimensionality, integrating different attention-driven methods. We trained the algorithm using DrugAge database. Our comprehensive analyses demonstrate that 64-bit Attention-ElixirFP maintains high predictive accuracy and F1 score while minimizing computational cost. Using ElixirSeeker to screen external compound databases, we identified a number of promising candidate anti-aging drugs. We tested top 6 hits and found that 4 of these compounds extend the lifespan of Caenorhabditis elegans, including Polyphyllin , Medrysone, Thymoquinone and Medrysone. This study illustrates that attention-driven fusion of fingerprints maximizes the learning of molecular activity features, providing a novel approach for high-throughput machine learning discovery of anti-aging molecules.
]]></description>
<dc:creator>Pan, Y.</dc:creator>
<dc:creator>Cai, H.</dc:creator>
<dc:creator>Ye, F.</dc:creator>
<dc:creator>Xu, W.</dc:creator>
<dc:creator>Huang, Z.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Gong, Y.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Ezemaduka, A. N.</dc:creator>
<dc:creator>Gao, S.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Ning, J.</dc:creator>
<dc:creator>Xian, B.</dc:creator>
<dc:date>2024-09-13</dc:date>
<dc:identifier>doi:10.1101/2024.09.08.611839</dc:identifier>
<dc:title><![CDATA[ElixirSeeker: A Machine Learning Framework Utilizing Attention-Driven Fusion of Molecular Fingerprints for the Discovery of Anti-Aging Compounds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.09.612077v1?rss=1">
<title>
<![CDATA[
Leukemia-derived apelin selects endothelial niche clones to promote tumorigenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.09.612077v1?rss=1</link>
<description><![CDATA[
Hematopoietic stem cells are regulated by endothelial and mesenchymal stromal cells in the marrow niche1-3. Leukemogenesis was long believed to be solely driven by genetic perturbations in hematopoietic cells but introduction of genetic mutations in the microenvironment demonstrated the ability of niche cells to drive disease progression4-8. The mechanisms by which the stem cell niche induces leukemia remain poorly understood. Here, using cellular barcoding in zebrafish, we found that clones of niche endothelial and stromal cells are significantly expanded in leukemic marrows. The pro-angiogenic peptide apelin secreted by leukemic cells induced sinusoidal endothelial cell clonal selection and transcriptional reprogramming towards an angiogenic state to promote leukemogenesis in vivo. Overexpression of apelin in normal hematopoietic stem cells led to clonal amplification of the niche endothelial cells and promotes clonal dominance of blood cells. Knock-out of apelin in leukemic zebrafish resulted in a significant reduction in disease progression. Our results demonstrate that leukemic cells remodel the clonal and transcriptional landscape of the marrow niche to promote leukemogenesis and provide a potential therapeutic opportunity for anti-apelin treatment.
]]></description>
<dc:creator>Baron, C. S.</dc:creator>
<dc:creator>Mitchell, O.</dc:creator>
<dc:creator>Avagyan, S.</dc:creator>
<dc:creator>Menard, R.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Robertson, A. L.</dc:creator>
<dc:creator>Potluri, R.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Madelaine, R.</dc:creator>
<dc:creator>McKenna, A.</dc:creator>
<dc:creator>Zon, L. I.</dc:creator>
<dc:date>2024-09-13</dc:date>
<dc:identifier>doi:10.1101/2024.09.09.612077</dc:identifier>
<dc:title><![CDATA[Leukemia-derived apelin selects endothelial niche clones to promote tumorigenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.10.612298v1?rss=1">
<title>
<![CDATA[
BST2 induces vascular smooth muscle cell plasticity and phenotype switching during cancer progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.10.612298v1?rss=1</link>
<description><![CDATA[
BackgroundSmooth muscle cell (SMC) plasticity and phenotypic switching play prominent roles in the pathogenesis of multiple diseases, but their role in tumorigenesis is unknown. We investigated whether and how SMC diversity and plasticity plays a role in tumor angiogenesis and the tumor microenvironment.

Methods and ResultsWe use SMC-specific lineage-tracing mouse models and single cell RNA sequencing to observe the phenotypic diversity of SMCs participating in tumor vascularization. We find that a significant proportion of SMCs adopt a phenotype traditionally associated with macrophage-like cells. These cells are transcriptionally similar to  resolution phase M2b macrophages, which have been described to have a role in inflammation resolution. Computationally predicted by the ligand-receptor algorithm CellChat, signaling from BST2 on the surface of tumor cells to PIRA2 on SMCs promote this phenotypic transition; in vitro SMC assays demonstrate upregulation of macrophage transcriptional programs, and increased proliferation, migration, and phagocytic ability when exposed to BST2. Knockdown of BST2 in the tumor significantly decreases the transition towards a macrophage-like phenotype, and cells that do transition have a comparatively higher inflammatory signal typically associated with anti-tumor effect.

ConclusionAs BST2 is known to be a poor prognostic marker in multiple cancers where it is associated with an M2 macrophage-skewed TME, these studies suggest that phenotypically switched SMCs may have a previously unidentified role in this immunosuppressive milieu. Further translational work is needed to understand how this phenotypic switch could influence the response to anti-cancer agents and if targeted inhibition of SMC plasticity would be therapeutically beneficial.
]]></description>
<dc:creator>Bell, C.</dc:creator>
<dc:creator>Baylis, R.</dc:creator>
<dc:creator>Lopez, N.</dc:creator>
<dc:creator>Ma, W. F.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Bamezai, S.</dc:creator>
<dc:creator>Fu, C.</dc:creator>
<dc:creator>Kojima, Y.</dc:creator>
<dc:creator>Adkar, S.</dc:creator>
<dc:creator>Luo, L.</dc:creator>
<dc:creator>Miller, C.</dc:creator>
<dc:creator>Leeper, N. L.</dc:creator>
<dc:date>2024-09-14</dc:date>
<dc:identifier>doi:10.1101/2024.09.10.612298</dc:identifier>
<dc:title><![CDATA[BST2 induces vascular smooth muscle cell plasticity and phenotype switching during cancer progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.13.612389v1?rss=1">
<title>
<![CDATA[
Intracortical injection of immune checkpoint inhibitor promotes monocyte/macrophage infiltration and restores microglial function and neuronal activity in an AD mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.13.612389v1?rss=1</link>
<description><![CDATA[
Alzheimer's disease (AD) involves complex neuroimmune interactions. However, the role of immune checkpoint pathways in regulating the glial and neuronal functions of the AD brain remains unclear. This study aims to investigate how the brain-specific modulation of the PD-1/PD-L1 axis affects glial and neuronal function in an AD mouse model, which is characterized by the significant upregulation of microglial PD-1 and astrocytic PD-L1. A single intracortical administration of anti-PD-L1 antibody reshaped the glial microenvironment, which was accompanied by the restoration of impaired microglial process convergence. Astrocyte-specific Pd-l1 knockdown using the pSico system validated the essential role of astrocytic PD-L1 in enhancing microglial responses. Notably, the blockade of the PD-1/PD-L1 signaling pathway increased the microglial P2RY12 expression, a key marker of microglial homeostasis, which likely contributed to the restoration of neuronal hyperactivity. Collectively, these findings confirmed the potential of targeting the astrocyte-microglia PD-1/PD-L1 axis to mitigate AD pathology.
]]></description>
<dc:creator>Park, T.</dc:creator>
<dc:creator>Chang, L.</dc:creator>
<dc:creator>Chung, S. W.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Bae, S.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:creator>Kim, Y. H.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Kim, H.-J.</dc:creator>
<dc:creator>Kwon, H.-K.</dc:creator>
<dc:creator>Suh, M.</dc:creator>
<dc:date>2024-09-15</dc:date>
<dc:identifier>doi:10.1101/2024.09.13.612389</dc:identifier>
<dc:title><![CDATA[Intracortical injection of immune checkpoint inhibitor promotes monocyte/macrophage infiltration and restores microglial function and neuronal activity in an AD mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.13.612783v1?rss=1">
<title>
<![CDATA[
Learning-related oscillatory dynamics in the human cortico-basal ganglia network during motor sequence initiation in Parkinson's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.13.612783v1?rss=1</link>
<description><![CDATA[
Learning dexterous motor sequences is crucial to autonomy and quality of life but can be altered in Parkinsons Disease (PD). Learning involves optimizing pre-movement planning (preplanning) of multiple sequence elements to reduce computational overhead during active movement. However, it is unclear which brain regions mediate preplanning or how this process evolves with learning. Recording cortico-basal ganglia field potentials during a multi-day typing task in four individuals with PD, we found evidence for network-wide multi-element preplanning that improved with learning, facilitated by functional connectivity. In both cortex and basal ganglia, pre-movement gamma ({gamma}, 30-250 Hz) activity, historically linked to population spiking, distinguished between future action sequences and became increasingly predictive with learning. For motor cortex {gamma}, this increase was tied to learning-related cross-frequency coupling led by cortically-driven network delta ({delta}, 0.5-4 Hz) synchrony. More generally, coordinated network {delta} supported a complex pattern of learning-driven cross-frequency couplings within and between cortex and basal ganglia, including striatal lead of cortical beta ({beta}, 12-30 Hz) activity, reflecting the specialized roles of these brain regions in motor preparation. In contrast, impaired learning was characterized by practice-driven decreases in {gamma}s predictive value, limited cross-frequency coupling and absent network {delta} synchrony, with network dynamics possibly altered by pathologically high inter-basal ganglia {delta} synchrony. These results suggest that cortically-led {delta} phase coordination optimized cortico-basal ganglia multi-element preplanning through enhanced recruitment of higher-frequency neural activity. Neurostimulation that enhances cortico-basal ganglia {delta} synchrony may thus hold potential for improving skilled fine motor control in PD.
]]></description>
<dc:creator>Presbrey, K. N.</dc:creator>
<dc:creator>Wozny, T. A.</dc:creator>
<dc:creator>Louie, K. H.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:creator>Starr, P. A.</dc:creator>
<dc:creator>Abbasi-Asl, R.</dc:creator>
<dc:creator>Wang, D. D.</dc:creator>
<dc:date>2024-09-15</dc:date>
<dc:identifier>doi:10.1101/2024.09.13.612783</dc:identifier>
<dc:title><![CDATA[Learning-related oscillatory dynamics in the human cortico-basal ganglia network during motor sequence initiation in Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.12.612619v1?rss=1">
<title>
<![CDATA[
Intravital imaging of pulmonary lymphatics in inflammation and metastatic cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.12.612619v1?rss=1</link>
<description><![CDATA[
Intravital microscopy has enabled the study of immune dynamics in the pulmonary microvasculature, but many key events remain unseen because they occur in deeper lung regions. We therefore developed a technique for stabilized intravital imaging of bronchovascular cuffs and collecting lymphatics surrounding pulmonary veins in mice. Intravital imaging of pulmonary lymphatics revealed ventilation-dependence of steady-state lung lymph flow and ventilation-independent lymph flow during inflammation. We imaged the rapid exodus of migratory dendritic cells through lung lymphatics following inflammation and measured effects of pharmacologic and genetic interventions targeting chemokine signaling. Intravital imaging also captured lymphatic immune surveillance of lung-metastatic cancers and lymphatic metastasis of cancer cells. To our knowledge, this is the first imaging of lymph flow and leukocyte migration through intact pulmonary lymphatics. This approach will enable studies of protective and maladaptive processes unfolding within the lungs and in other previously inaccessible locations.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=199 HEIGHT=200 SRC="FIGDIR/small/612619v1_ufig1.gif" ALT="Figure 1">
View larger version (64K):
org.highwire.dtl.DTLVardef@66dd00org.highwire.dtl.DTLVardef@1373576org.highwire.dtl.DTLVardef@1970dfeorg.highwire.dtl.DTLVardef@17c15c_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Cleary, S. J.</dc:creator>
<dc:creator>Qiu, L.</dc:creator>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>Baluk, P.</dc:creator>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Serwas, N. K.</dc:creator>
<dc:creator>Cyster, J. G.</dc:creator>
<dc:creator>McDonald, D. M.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Looney, M. R.</dc:creator>
<dc:date>2024-09-17</dc:date>
<dc:identifier>doi:10.1101/2024.09.12.612619</dc:identifier>
<dc:title><![CDATA[Intravital imaging of pulmonary lymphatics in inflammation and metastatic cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.16.613379v1?rss=1">
<title>
<![CDATA[
Quantitative Comparison of Monomeric StayGold Variants Using Protein Nanocages in Living Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.16.613379v1?rss=1</link>
<description><![CDATA[
To standardize comparison of fluorescent proteins and independently determine which monomeric StayGold variant is best for live microscopy, we analyzed fluorescent protein tagged I3-01 peptides that self-assemble into stable sixty subunit dodecahedrons inside live cells. We find mStayGold is 3-fold brighter and 3-fold more photostable compared with EGFP and superior to other monomeric variants in mammalian cytoplasm. In addition, analysis of intracellular nanocage diffusion confirms the monomeric nature of mStayGold.
]]></description>
<dc:creator>Viola, G.</dc:creator>
<dc:creator>Jacobs, K. A.</dc:creator>
<dc:creator>Lemiere, J.</dc:creator>
<dc:creator>Kutys, M. L.</dc:creator>
<dc:creator>Wittmann, T.</dc:creator>
<dc:date>2024-09-17</dc:date>
<dc:identifier>doi:10.1101/2024.09.16.613379</dc:identifier>
<dc:title><![CDATA[Quantitative Comparison of Monomeric StayGold Variants Using Protein Nanocages in Living Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.17.613524v1?rss=1">
<title>
<![CDATA[
A dopaminergic basis of behavioral control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.17.613524v1?rss=1</link>
<description><![CDATA[
Both goal-directed and automatic processes shape human behavior, but these processes often conflict. Behavioral control is the decision about which process guides behavior. Despite the importance of behavioral control for adaptive decision-making, its neural mechanisms remain unclear. Critically, it is unknown if there are mechanisms for behavioral control that are distinct from those supporting the formation of goal-relevant knowledge. We performed deep phenotyping of individual dopamine system function by combining multiple PET scans, fMRI, and dopaminergic drug administration in a within-subject, double-blind, placebo-controlled design. Subjects performed a rule-based response time task, with goal-directed and automatic decision-making operationalized as model-based and model-free influences on behavior. We found a double dissociation between two aspects of ventral striatal dopamine physiology: D2/3 receptor availability and dopamine synthesis capacity. Convergent and causal evidence indicated that D2/3 receptors regulate behavioral control by enhancing model-based and blunting model-free influences on behavior but do not affect model-based knowledge formation. In contrast, dopamine synthesis capacity was linked to the formation of model-based knowledge but not behavioral control. D2/3 receptors also modulated frontostriatal functional connectivity, suggesting they regulate behavioral control by gating prefrontal inputs to the striatum. These results identify central mechanisms underlying individual and state differences in behavioral control and point to striatal D2/3 receptors as targets for interventions for improving goal-directed behavior.
]]></description>
<dc:creator>Ballard, I. C.</dc:creator>
<dc:creator>Furman, D. J.</dc:creator>
<dc:creator>Berry, A. S.</dc:creator>
<dc:creator>White, R. L.</dc:creator>
<dc:creator>Jagust, W. J.</dc:creator>
<dc:creator>Kayser, A. S.</dc:creator>
<dc:creator>D'Esposito, M.</dc:creator>
<dc:date>2024-09-17</dc:date>
<dc:identifier>doi:10.1101/2024.09.17.613524</dc:identifier>
<dc:title><![CDATA[A dopaminergic basis of behavioral control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.17.613377v1?rss=1">
<title>
<![CDATA[
Engineered Receptors for Soluble Cell-to-Cell Communication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.17.613377v1?rss=1</link>
<description><![CDATA[
Despite recent advances in mammalian synthetic biology, there remains a lack of modular synthetic receptors that can robustly respond to soluble ligands and in turn activate cellular functions. Such receptors would have extensive clinical potential to regulate the activity of engineered therapeutic cells, but to date only receptors against cell surface targets have approached clinical translation. To address this gap, we developed a receptor architecture based on synNotch, called SyNthetic Intramembrane Proteolysis Receptor (SNIPR), that has the added ability to be activated by soluble ligands, both natural and synthetic, with remarkably low baseline activity and high fold activation. SNIPRs are able to access an endocytic, pH-dependent cleavage mechanism to achieve soluble ligand sensing, in addition to employing a canonical-like pathway for detecting surface-bound ligands. We demonstrate the therapeutic capabilities of the receptor platform by localizing the activity of CAR T-cells to solid tumors where soluble disease-associated factors are expressed, bypassing the major hurdle of on-target off-tumor toxicity in bystander organs. We further applied the SNIPR platform to engineer fully synthetic signaling networks between cells orthogonal to natural signaling pathways, expanding the scope of synthetic biology. Our design framework enables cellular communication and environmental interactions, extending the capabilities of synthetic cellular networking in clinical and research contexts.
]]></description>
<dc:creator>Piraner, D. I.</dc:creator>
<dc:creator>Abedi, M. H.</dc:creator>
<dc:creator>Duran Gonzalez, M. J.</dc:creator>
<dc:creator>Chazin-Gray, A.</dc:creator>
<dc:creator>Zhu, I.</dc:creator>
<dc:creator>Ravindran, P. T.</dc:creator>
<dc:creator>Schlichthaerle, T.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:creator>Roybal, K. T.</dc:creator>
<dc:date>2024-09-18</dc:date>
<dc:identifier>doi:10.1101/2024.09.17.613377</dc:identifier>
<dc:title><![CDATA[Engineered Receptors for Soluble Cell-to-Cell Communication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.13.612924v1?rss=1">
<title>
<![CDATA[
M2 Macrophage Exosomes Reverse Cardiac Functional Decline in Mice with Diet-Induced Myocardial Infarction by Suppressing Type 1 Interferon Signaling in Myeloid Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.13.612924v1?rss=1</link>
<description><![CDATA[
Effective treatment strategies to alleviate heart failure that develops as a consequence of myocardial infarction (MI) remain an unmet need in cardiovascular medicine. In this study, we uncovered that exosomes produced by human THP-1 macrophages cultured with the cytokine IL-4 (THP1-IL4-exo), reverse cardiac functional decline in mice that develop MI as a consequence of diet-induced occlusive coronary atherosclerosis. Therapeutic benefits of THP1-IL4-exo stem from their ability to reprogram circulating Ly-6Chi monocytes into an M2-like phenotype and suppress Type 1 Interferon signaling in myeloid cells within the bone marrow, the circulation, and cardiac tissue. Collectively, these benefits suppress myelopoiesis, myeloid cell recruitment to cardiac tissue, and preserve populations of resident cardiac macrophages that together mitigate cardiac inflammation, adverse ventricular remodeling, and heart failure. Our findings introduce THP1-IL4-exo, one form of M2-macrophage exosomes, as novel therapeutics to preserve cardiac function subsequent to MI.
]]></description>
<dc:creator>Ng, M.</dc:creator>
<dc:creator>Gao, A. S.</dc:creator>
<dc:creator>Phu, T. A.</dc:creator>
<dc:creator>Vu, N. K.</dc:creator>
<dc:creator>Raffai, R.</dc:creator>
<dc:date>2024-09-19</dc:date>
<dc:identifier>doi:10.1101/2024.09.13.612924</dc:identifier>
<dc:title><![CDATA[M2 Macrophage Exosomes Reverse Cardiac Functional Decline in Mice with Diet-Induced Myocardial Infarction by Suppressing Type 1 Interferon Signaling in Myeloid Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.16.613176v1?rss=1">
<title>
<![CDATA[
Real-space heterogeneous reconstruction, refinement, and disentanglement of CryoEM conformational states with HetSIREN 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.16.613176v1?rss=1</link>
<description><![CDATA[
Single-particle analysis by Cryo-electron microscopy (CryoEM) provides direct access to the conformation of each macromolecule. However, the images signal-to-noise ratio is low, and some form of classification is usually performed at the image processing level to allow structural modeling. Classical classification methods imply the existence of a discrete number of structural conformations. However, new heterogeneity algorithms introduce a novel reconstruction paradigm, where every state is represented by a lower number of particles, potentially just one, allowing the estimation of conformational landscapes representing the different structural states a biomolecule explores. In this work, we present a novel deep learning-based method called HetSIREN. HetSIREN can fully reconstruct or refine a CryoEM volume in real space based on the structural information summarized in a conformational latent space. The unique characteristics that set HetSIREN apart start with the definition of the approach as a real space-based only method, a fact that allows spatially focused analysis, but also the introduction of a novel network architecture specifically designed to make use of meta-sinusoidal activations, with proven high analytics capacities. Continuing with innovations, HetSIREN can also refine the pose parameters of the images at the same time that it conditions the network with prior information/constraints on the maps, such as Total Variation and L1 denoising, ultimately yielding cleaner volumes with high-quality structural features. Finally, but very importantly, HetSIREN addresses one of the most confusing issues in heterogeneity analysis, as it is the fact that real structural heterogeneity estimation is entangled with pose estimation (and to a lesser extent with CTF estimation), in this way, HetSIREN introduces a novel encoding architecture able to decouple pose and CTF information from the conformational landscape, resulting in more accurate and interpretable conformational latent spaces. We present results on computer-simulated data, public data from EMPIAR, and data from experimental systems currently being studied in our laboratories. An important finding is the sensitivity of the structure and dynamics of the SARS-CoV-2 Spike protein on the storage temperature.
]]></description>
<dc:creator>Herreros, D.</dc:creator>
<dc:creator>Mata, C. P.</dc:creator>
<dc:creator>Noddings, C.</dc:creator>
<dc:creator>Irene, D.</dc:creator>
<dc:creator>Krieger, J. M.</dc:creator>
<dc:creator>Agard, D.</dc:creator>
<dc:creator>Tsai, M.-D.</dc:creator>
<dc:creator>Sorzano, C. O. S.</dc:creator>
<dc:creator>Carazo, J. M.</dc:creator>
<dc:date>2024-09-19</dc:date>
<dc:identifier>doi:10.1101/2024.09.16.613176</dc:identifier>
<dc:title><![CDATA[Real-space heterogeneous reconstruction, refinement, and disentanglement of CryoEM conformational states with HetSIREN]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.17.611698v1?rss=1">
<title>
<![CDATA[
The Gq/11 family of Gα subunits is necessary and sufficient for lower jaw development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.17.611698v1?rss=1</link>
<description><![CDATA[
Vertebrate jaw development is coordinated by highly conserved ligand-receptor systems such as the peptide ligand Endothelin 1 (Edn1) and Endothelin receptor type A (Ednra), which are required for patterning of lower jaw structures. The Edn1/Ednra signaling pathway establishes the identity of lower jaw progenitor cells by regulating expression of numerous patterning genes, but the intracellular signaling mechanisms linking receptor activation to gene regulation remain poorly understood. As a first step towards elucidating this mechanism, we examined the function of the Gq/11 family of G subunits in zebrafish using pharmacological inhibition and genetic ablation of Gq/11 activity and transgenic induction of a constitutively active Gq protein in edn1-/- embryos. Genetic loss of Gq/11 activity fully recapitulated the edn1-/- phenotype, with genes encoding G11 being most essential. Furthermore, inducing Gq activity in edn1-/-embryos not only restored Edn1/Ednra-dependent jaw structures and gene expression signatures but also caused homeosis of the upper jaw structure into a lower jaw-like structure. These results indicate that Gq/11 is necessary and sufficient to mediate the lower jaw patterning mechanism for Ednra in zebrafish.

Summary statementGq/11 is the signaling mediator downstream of Endothelin 1 and Endothelin Receptor Type A that drives tissue patterning for all lower jaw structures in zebrafish.
]]></description>
<dc:creator>Kanai, S. M.</dc:creator>
<dc:creator>Garcia, C. R.</dc:creator>
<dc:creator>Augustus, M. R.</dc:creator>
<dc:creator>Sharafeldeen, S. A.</dc:creator>
<dc:creator>Brooks, E. P.</dc:creator>
<dc:creator>Sucharov, J.</dc:creator>
<dc:creator>Lencer, E. S.</dc:creator>
<dc:creator>Nichols, J. T.</dc:creator>
<dc:creator>Clouthier, D. E.</dc:creator>
<dc:date>2024-09-19</dc:date>
<dc:identifier>doi:10.1101/2024.09.17.611698</dc:identifier>
<dc:title><![CDATA[The Gq/11 family of Gα subunits is necessary and sufficient for lower jaw development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.17.613111v1?rss=1">
<title>
<![CDATA[
An integrated view of the structure and function of the human 4D nucleome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.17.613111v1?rss=1</link>
<description><![CDATA[
The dynamic three-dimensional (3D) organization of the human genome (the "4D Nucleome") is closely linked to genome function. Here, we integrate a wide variety of genomic data generated by the 4D Nucleome Project to provide a detailed view of human 3D genome organization in widely used embryonic stem cells (H1-hESCs) and immortalized fibroblasts (HFFc6). We provide extensive benchmarking of 3D genome mapping assays and integrate these diverse datasets to annotate spatial genomic features across scales. The data reveal a rich complexity of chromatin domains and their sub-nuclear positions, and over one hundred thousand structural loops and promoter-enhancer interactions. We developed 3D models of population-based and individual cell-to-cell variation in genome structure, establishing connections between chromosome folding, nuclear organization, chromatin looping, gene transcription, and DNA replication. We demonstrate the use of computational methods to predict genome folding from DNA sequence, uncovering potential effects of genetic variants on genome structure and function. Together, this comprehensive analysis contributes insights into human genome organization and enhances our understanding of connections between the regulation of genome function and 3D genome organization in general.
]]></description>
<dc:creator>The 4D Nucleome Consortium,</dc:creator>
<dc:creator>Dekker, J.</dc:creator>
<dc:creator>Oksuz, B. A.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Minsk, M. K.</dc:creator>
<dc:creator>Kuang, S.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Gibcus, J. H.</dc:creator>
<dc:creator>Krietenstein, N.</dc:creator>
<dc:creator>Rando, O.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Kukalev, A.</dc:creator>
<dc:creator>Willemin, A.</dc:creator>
<dc:creator>Winick-Ng, W.</dc:creator>
<dc:creator>Kempfer, R.</dc:creator>
<dc:creator>Pombo, A.</dc:creator>
<dc:creator>Yu, M.</dc:creator>
<dc:creator>Kumar, P.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Belmont, A. S.</dc:creator>
<dc:creator>Sasaki, T.</dc:creator>
<dc:creator>van Schaik, T.</dc:creator>
<dc:creator>Brueckner, L.</dc:creator>
<dc:creator>Peric-Hupkes, D.</dc:creator>
<dc:creator>van Steensel, B.</dc:creator>
<dc:creator>Wang, P.</dc:creator>
<dc:creator>Chai, H.</dc:creator>
<dc:creator>Kim, M.</dc:creator>
<dc:creator>Ruan, Y.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Quinodoz, S. A.</dc:creator>
<dc:creator>Bhat, P.</dc:creator>
<dc:creator>Guttman, M.</dc:creator>
<dc:creator>Zhao, W.</dc:creator>
<dc:creator>Chien, S.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Venev, S. V.</dc:creator>
<dc:creator>Plewczynski, D.</dc:creator>
<dc:creator>Irastorza Azcarate, I.</dc:creator>
<dc:creator>Szabo, D.</dc:creator>
<dc:creator>Thieme, C. J.</dc:creator>
<dc:creator>Szczepinska,</dc:creator>
<dc:date>2024-09-19</dc:date>
<dc:identifier>doi:10.1101/2024.09.17.613111</dc:identifier>
<dc:title><![CDATA[An integrated view of the structure and function of the human 4D nucleome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.20.612365v1?rss=1">
<title>
<![CDATA[
Preventing Inappropriate Signals Pre- and Post-Ligand Perception by a Toggle-Switch Mechanism of ERECTA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.20.612365v1?rss=1</link>
<description><![CDATA[
Dynamic control of signaling events requires swift regulation of receptors at an active state. By focusing on Arabidopsis ERECTA (ER) receptor kinase, which perceives peptide ligands to control multiple developmental processes, we report a mechanism preventing inappropriate receptor activity. The ER C-terminal tail (ER_CT) functions as an autoinhibitory domain: its removal confers higher kinase activity and hyperactivity during inflorescence and stomatal development. ER_CT is required for the binding of a receptor kinase inhibitor, BKI1, and two U-box E3 ligases PUB30 and PUB31 that inactivate activated ER. We further identify ER_CT as a phosphodomain trans-phosphorylated by the co-receptor BAK1. The phosphorylation impacts the tail structure, likely releasing from autoinhibition. The phosphonull version enhances BKI1 association, whereas the phosphomimetic version promotes PUB30/31 association. Thus, ER_CT acts as an off-on-off toggle switch, facilitating the release of BKI1 inhibition, enabling signal activation, and swiftly turning over the receptors afterwards. Our results elucidate a mechanism fine-tuning receptor signaling via a phosphoswitch module, keeping the receptor at a low basal state and ensuring the robust yet transient activation upon ligand perception.

SignificanceCells perceive and process external signals through their cell-surface receptors, whose activity must be tightly maintained to prevent the spread of misinformation. How do plant cells prevent the inappropriate receptor activity? We identify a structural module within the C-terminal tail of the ERECTA (ER_CT), that inhibits the receptor pre- and post-signal activation. The ER_CT comprises of a linker and an -helix. Before activation, ER_CT is autoinhibitory and associates with an inhibitory protein. Ligand perception triggers the transphosphorylation of ER_CT by the co-receptor, which then recruits a degradation machinery to swiftly turn over the activated receptor. Thus, we reveal an off-on-off toggle switch mechanism that finely adjusts the activity of the plant receptor, enabling the precise control over cell signaling.
]]></description>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Maes, M.</dc:creator>
<dc:creator>Cochran, A. M.</dc:creator>
<dc:creator>Avila, J. R.</dc:creator>
<dc:creator>Derbyshire, P.</dc:creator>
<dc:creator>Sklenar, J.</dc:creator>
<dc:creator>Haas, K. M.</dc:creator>
<dc:creator>Villen, J.</dc:creator>
<dc:creator>Menke, F.</dc:creator>
<dc:creator>Torii, K. U.</dc:creator>
<dc:date>2024-09-21</dc:date>
<dc:identifier>doi:10.1101/2024.09.20.612365</dc:identifier>
<dc:title><![CDATA[Preventing Inappropriate Signals Pre- and Post-Ligand Perception by a Toggle-Switch Mechanism of ERECTA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.20.613821v1?rss=1">
<title>
<![CDATA[
Extinction of innate floral preferences in the pollinator Eristalis tenax 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.20.613821v1?rss=1</link>
<description><![CDATA[
Innate behaviors allow solitary animals to complete essential tasks in the absence of social learning. However, we know little about the degree to which ecologically relevant innate preferences can change. The hoverfly Eristalis tenax, a solitary generalist pollinator, is an ideal model for studying innate behavior in a naturalistic context because its survival depends on the innate ability to identify flowers. Innate behavior in E. tenax has previously been considered inalterable, but we hypothesized that E. tenax can modulate their innate behavior after training in a multimodal sensory context, in contrast to the prior work that employed unimodal sensory cues. To test this, we examined if E. tenax can extinguish an innate proboscis extension response (PER) to a multimodal floral object after undergoing aversive conditioning with quinine, and if flies can acquire PER to an innately unattractive object using sucrose as reinforcement. Finally, we assessed long-term memory retention. Here, we report a complete extinction of the proboscis extension response (PER) to an innately attractive floral object following aversive training. E. tenax can also acquire PER to an innately unattractive object after appetitive training. Flies can retain these memories for days after training, and aversive memories last longer than appetitive memories. Our results contrast with literature stating that innate preferences cannot be extinguished in E. tenax. This could be because our study uses multimodal objects instead of the unimodal stimuli used in previous work. Ultimately, these findings improve our understanding of how animals navigate the uncertainties of dynamic objects in the natural world.
]]></description>
<dc:creator>Mishra, A.</dc:creator>
<dc:creator>Rajan, D. H.</dc:creator>
<dc:creator>Sharan, M.</dc:creator>
<dc:creator>Gharpure, G.</dc:creator>
<dc:creator>Olsson, S.</dc:creator>
<dc:date>2024-09-21</dc:date>
<dc:identifier>doi:10.1101/2024.09.20.613821</dc:identifier>
<dc:title><![CDATA[Extinction of innate floral preferences in the pollinator Eristalis tenax]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.18.613749v1?rss=1">
<title>
<![CDATA[
BCR ligation selectively inhibits IgE class switch recombination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.18.613749v1?rss=1</link>
<description><![CDATA[
Mechanisms that restrict class switch recombination (CSR) to IgE limit the subsequent production of IgE antibodies and therefore the development of allergic disease. Mice with impaired B cell receptor (BCR) signaling have significantly increased IgE responses, consistent with a role for BCR signaling in IgE regulation. While prior work focused on BCR signaling in IgE-expressing cells to explain these findings, it has been reported that BCR signaling can reduce CSR. Therefore, we investigated the possibility that IgE CSR might be particularly sensitive to inhibition by BCR signaling in unswitched B cells. We found that immunization of mice with high-affinity antigen resulted in reduced representation of IgE-expressing cells among germinal center B cells and plasma cells relative to a low-affinity antigen. Mechanistic experiments with cultured mouse B cells demonstrated that BCR ligands selectively inhibited IgE CSR in a dose-, affinity-, and avidity-dependent manner. Signaling via Syk was required for the inhibition of IgE CSR following BCR stimulation, whereas inhibition of the PI3K subunit p110{delta} increased IgE CSR independently of BCR ligation. The inhibition of IgE CSR by BCR ligands synergized with IL-21 or TGF{beta}1. BCR ligation also inhibited CSR to IgE in human tonsillar B cells, and this inhibition was also synergistic with IL-21. These findings establish that IgE CSR is uniquely susceptible to inhibition by BCR signaling in mouse and human B cells, with important implications for the regulation and pathogenesis of allergic disease.
]]></description>
<dc:creator>Wade-Vallance, A. K.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Libang, J. B.</dc:creator>
<dc:creator>Krishnapura, A. R.</dc:creator>
<dc:creator>Jung, J. B.</dc:creator>
<dc:creator>Matcham, E. W.</dc:creator>
<dc:creator>Robinson, M. J.</dc:creator>
<dc:creator>Allen, C. D. C.</dc:creator>
<dc:date>2024-09-22</dc:date>
<dc:identifier>doi:10.1101/2024.09.18.613749</dc:identifier>
<dc:title><![CDATA[BCR ligation selectively inhibits IgE class switch recombination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.22.614349v1?rss=1">
<title>
<![CDATA[
The HIV-1 Nuclear Export Complex Reveals the Role of RNA in Crm1 Cargo Recognition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.22.614349v1?rss=1</link>
<description><![CDATA[
Crm1 is a highly conserved nuclear exportin that transports >1000 human proteins including ribonucleoprotein (RNP) complexes. The interface between Crm1 and RNP cargos is unknown. The HIV regulatory protein, Rev, was one of the first identified cargos for Crm1 and contains a prototypic nuclear export sequence (NES). We present the cryo-electron microscopy structure of the HIV-1 nuclear export complex (Crm1/Ran-GTP and the Rev/RRE RNP). Rev binds at a previously unseen protein-protein binding site that stabilizes a unique Crm1 dimer and positions two NESs within the Crm1 dimer. The orientation of Rev binding positions the RRE within a charged pocket on the inside of the Crm1 toroid, mediating direct RNA-Ran-GTP contacts, highlighting the significant role of the RRE in the interaction. Structure based mutations, combined with cell-based assays, show that Crm1 has multiple distinct cargo recognition sites and explains how Crm1 can recognize a diverse range of protein and RNP cargos.
]]></description>
<dc:creator>Smith, A. M.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Velarde, A.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Frankel, A. D.</dc:creator>
<dc:date>2024-09-22</dc:date>
<dc:identifier>doi:10.1101/2024.09.22.614349</dc:identifier>
<dc:title><![CDATA[The HIV-1 Nuclear Export Complex Reveals the Role of RNA in Crm1 Cargo Recognition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.20.613996v1?rss=1">
<title>
<![CDATA[
Expanding Automated Multiconformer Ligand Modeling to Macrocycles and Fragments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.20.613996v1?rss=1</link>
<description><![CDATA[
Small molecule ligands exhibit a diverse range of conformations in solution. Upon binding to a target protein, this conformational diversity is reduced. However, ligands can retain some degree of conformational flexibility even when bound to a receptor. In the Protein Data Bank (PDB), a small number of ligands have been modeled with distinct alternative conformations that are supported by macromolecular X-ray crystallography density maps. However, the vast majority of structural models are fit to a single ligand conformation, potentially ignoring the underlying conformational heterogeneity present in the sample. We previously developed qFit-ligand to sample diverse ligand conformations and to select a parsimonious ensemble consistent with the density. While this approach indicated that many ligands populate alternative conformations, limitations in our sampling procedures often resulted in non-physical conformations and could not model complex ligands like macrocycles. Here, we introduce several improvements to qFit-ligand, including integrating RDKit for stochastic conformational sampling. This new sampling method greatly enriches low energy conformations of small molecules and macrocycles. We further extended qFit-ligand to identify alternative conformations in PanDDA-modified density maps from high throughput X-ray fragment screening experiments, as well as single-particle cryo-electron microscopy (cryo-EM) density maps. The new version of qFit-ligand improves fit to electron density and reduces torsional strain relative to deposited single conformer models and our prior version of qFit-ligand. These advances enhance the analysis of residual conformational heterogeneity present in ligand-bound structures, which can provide important insights for the rational design of therapeutic agents.
]]></description>
<dc:creator>Flowers, J.</dc:creator>
<dc:creator>Echols, N.</dc:creator>
<dc:creator>Correy, G.</dc:creator>
<dc:creator>Jaishankar, P.</dc:creator>
<dc:creator>Togo, T.</dc:creator>
<dc:creator>Renslo, A. R.</dc:creator>
<dc:creator>van den Bedem, H.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Wankowicz, S. A.</dc:creator>
<dc:date>2024-09-23</dc:date>
<dc:identifier>doi:10.1101/2024.09.20.613996</dc:identifier>
<dc:title><![CDATA[Expanding Automated Multiconformer Ligand Modeling to Macrocycles and Fragments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.20.614197v1?rss=1">
<title>
<![CDATA[
Tau P301S Transgenic Mice Develop Gait and Eye Movement Impairments That Mimic Progressive Supranuclear Palsy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.20.614197v1?rss=1</link>
<description><![CDATA[
Progressive supranuclear palsy (PSP) is a neurodegenerative disorder with an estimated prevalence of 5-7 people in 100,000. Clinically characterized by impairments in gait, balance, and eye movements, as well as aggregated Tau pathology, PSP leads to death in approximately 5-8 years. No disease-modifying treatments are currently available. The contribution of Tau pathology to the symptoms of patients with PSP is poorly understood, in part due to lack of a rodent model that recapitulates characteristic aspects of PSP. Here, we assessed the hTau.P301S mouse for key clinical features of PSP, finding progressive impairments in balance and gait coordination. Additionally, we found impairments in fast vertical eye movements, one of the most distinctive features of PSP. Across animals, we found that Tau pathology in motor control regions correlated with motor deficits. These findings highlight the utility of the hP301S mouse in modeling key aspects of PSP.
]]></description>
<dc:creator>Creed, R. B.</dc:creator>
<dc:creator>Harris, S. C.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>du Lac, S.</dc:creator>
<dc:creator>Zee, D. S.</dc:creator>
<dc:creator>Dunn, F. A.</dc:creator>
<dc:creator>Bouvier, G.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2024-09-23</dc:date>
<dc:identifier>doi:10.1101/2024.09.20.614197</dc:identifier>
<dc:title><![CDATA[Tau P301S Transgenic Mice Develop Gait and Eye Movement Impairments That Mimic Progressive Supranuclear Palsy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.21.613766v1?rss=1">
<title>
<![CDATA[
Intracellular diffusion in the cytoplasm increases with cell size in fission yeast 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.21.613766v1?rss=1</link>
<description><![CDATA[
Diffusion in the cytoplasm can greatly impact cellular processes, yet regulation of macromolecular diffusion remains poorly understood. There is increasing evidence that cell size affects the density and macromolecular composition of the cytoplasm. Here, we studied whether cell size affects diffusion at the scale of macromolecules tens of microns in diameter. We analyzed the diffusive motions of intracellular genetically-encoded multimeric 40 nm nanoparticles (cytGEMs) in the cytoplasm of the fission yeast Schizosaccharomyces pombe. Using cell size mutants, we showed that cytGEMs diffusion coefficients decreased in smaller cells and increased in larger cells. This increase in diffusion in large cells may be due to a decrease in the DNA-to-Cytoplasm ratio, as diffusion was not affected in large multinucleate cytokinesis mutants. In investigating the underlying causes of altered cytGEMs diffusion, we found that the proteomes of large and small cells exhibited size-specific changes, including the sub-scaling of ribosomal proteins in large cells. Comparison with a similar dataset from human cells revealed that features of size-dependent proteome remodeling were conserved. These studies demonstrate that cell size is an important parameter in determining the biophysical properties and the composition of the cytoplasm.
]]></description>
<dc:creator>Tan, C.</dc:creator>
<dc:creator>Lanz, M.</dc:creator>
<dc:creator>Swaffer, M.</dc:creator>
<dc:creator>Skotheim, J.</dc:creator>
<dc:creator>Chang, F.</dc:creator>
<dc:date>2024-09-23</dc:date>
<dc:identifier>doi:10.1101/2024.09.21.613766</dc:identifier>
<dc:title><![CDATA[Intracellular diffusion in the cytoplasm increases with cell size in fission yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.21.614267v1?rss=1">
<title>
<![CDATA[
Super-enhancer profiling reveals ThPOK/ZBTB7B, a CD4+ cell lineage commitment factor, as a master regulator that restricts breast cancer cells to a luminal non-migratory phenotype 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.21.614267v1?rss=1</link>
<description><![CDATA[
Despite efforts to understand breast cancer biology, metastatic disease remains a clinical challenge. Identifying suppressors of breast cancer progression and mechanisms of transition to more invasive phenotypes could provide game changing therapeutic opportunities. Transcriptional deregulation is central to all malignancies, highlighted by the extensive reprogramming of regulatory elements that underlie oncogenic programs. Among these, super-enhancers (SEs) stand out due to their enrichment in genes controlling cancer hallmarks. To reveal novel breast cancer dependencies, we integrated the analysis of the SE landscape with master regulator activity inference for a series of breast cancer cell lines. As a result, we identified T-helper-inducing Poxviruses and Zinc-finger (POZ)/Kruppel-like factor (ThPOK, ZBTB7B), a CD4+ cell lineage commitment factor, as a breast cancer master regulator that is recurrently associated with a SE. ThPOK expression is highest in luminal breast cancer but is significantly reduced in the basal subtype. Manipulation of ThPOK levels in cell lines shows that its repressive function restricts breast cancer cells to an epithelial phenotype by suppressing the expression of genes involved in the epithelial-mesenchymal transition (EMT), WNT/{beta}-catenin target genes, and the pro-metastatic TGF{beta} pathway. Our study reveals ThPOK as a master transcription factor that restricts the acquisition of metastatic features in breast cancer cells.
]]></description>
<dc:creator>Arcuschin, C. D.</dc:creator>
<dc:creator>Kahrizi, K.</dc:creator>
<dc:creator>Sayaman, R. W.</dc:creator>
<dc:creator>DiBenedetto, C.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Salaberry, P. J.</dc:creator>
<dc:creator>Zakroui, O.</dc:creator>
<dc:creator>Schwarzer, C.</dc:creator>
<dc:creator>Scapozza, A.</dc:creator>
<dc:creator>Betancur, P.</dc:creator>
<dc:creator>Saba, J. D.</dc:creator>
<dc:creator>Coppe, J.-P.</dc:creator>
<dc:creator>Barcellos-Hoff, M. H.</dc:creator>
<dc:creator>Kappes, D.</dc:creator>
<dc:creator>van 't Veer, L.</dc:creator>
<dc:creator>Schor, I. E.</dc:creator>
<dc:creator>Munoz, D. P.</dc:creator>
<dc:date>2024-09-24</dc:date>
<dc:identifier>doi:10.1101/2024.09.21.614267</dc:identifier>
<dc:title><![CDATA[Super-enhancer profiling reveals ThPOK/ZBTB7B, a CD4+ cell lineage commitment factor, as a master regulator that restricts breast cancer cells to a luminal non-migratory phenotype]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.23.614410v1?rss=1">
<title>
<![CDATA[
Comprehensive proteolytic profiling of Aedes aegypti mosquito midgut extracts: Unraveling the blood meal protein digestion system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.23.614410v1?rss=1</link>
<description><![CDATA[
To sustain the gonotrophic cycle, the Aedes aegypti mosquito must acquire a blood meal from a human or other vertebrate host. However, in the process of blood feeding, the mosquito may facilitate the transmission of several bloodborne viral pathogens (e.g., dengue, Zika, and chikungunya). The blood meal is essential as it contains proteins that are digested into polypeptides and amino acid nutrients that are eventually used for egg production. These proteins are digested by several midgut proteolytic enzymes. As such, the female mosquitos reliance on blood may serve as a potential target for vector and viral transmission control. However, this strategy may prove to be challenging since midgut proteolytic activity is a complex process dependent on several exo- and endo-proteases. Therefore, to understand the complexity of Ae. aegypti blood meal digestion, we used Multiplex Substrate Profiling by Mass Spectrometry (MSP-MS) to generate global proteolytic profiles of sugar- and blood-fed midgut tissue extracts, along with substrate profiles of recombinantly expressed midgut proteases. Our results reveal a shift from high exoproteolytic activity in sugar-fed mosquitoes to an expressive increase in endoproteolytic activity in blood-fed mosquitoes. This approach allowed for the identification of 146 cleaved peptide bonds (by the combined 6 h and 24 h blood-fed samples) in the MSP-MS substrate library, and of these 146, 99 (68%) were cleaved by the five recombinant proteases evaluated. These reveal the individual contribution of each recombinant midgut protease to the overall blood meal digestion process of the Ae. aegypti mosquito. Further, our molecular docking simulations support the substrate specificity of each recombinant protease. Therefore, the present study provides key information of midgut proteases and the blood meal digestion process in mosquitoes, which may be exploited for the development of potential inhibitor targets for vector and viral transmission control strategies.

Author SummaryThe Aedes aegypti mosquito is a vector of viral pathogens that can be transmitted directly to humans. For instance, the transmission of dengue, Zika, or chikungunya viruses may happen during the Ae. aegypti acquisition of an infected blood meal. This blood meal is important for the anautogenous mosquito because without the digestion of blood proteins the mosquito will not obtain the necessary nutrients needed for egg production. After imbibing a blood meal, midgut digestive enzymes (proteases) are expressed and secreted into the lumen. To fully understand their roles in blood meal digestion, we used a special technique called Multiplex Substrate Profiling by Mass Spectrometry (MSP-MS). This method allows us to generate global proteolytic activity profiles of Ae. aegypti midgut tissue extracts that were fed with sugar or blood. In addition, we generated substrate cleavage profiles of recombinantly expressed midgut proteases allowing us to understand the enzyme preferences for blood proteins. Therefore, utilizing this approach, we found the contribution of each individual recombinant protease tested relative to the global activity profile of blood-fed midgut tissue extracts. This may be a starting point for the validation of midgut protease inhibition and the development of a new potential vector control strategy.
]]></description>
<dc:creator>O'Donoghue, A. J.</dc:creator>
<dc:creator>Lui, C.</dc:creator>
<dc:creator>Simington, C. J.</dc:creator>
<dc:creator>Montermoso, S.</dc:creator>
<dc:creator>Moreno-Galvez, E.</dc:creator>
<dc:creator>Serafim, M. S. M.</dc:creator>
<dc:creator>Burata, O. E.</dc:creator>
<dc:creator>Lucero, R. M.</dc:creator>
<dc:creator>Nguyen, J. T.</dc:creator>
<dc:creator>Fong, D.</dc:creator>
<dc:creator>Tran, K.</dc:creator>
<dc:creator>Millan, N.</dc:creator>
<dc:creator>Gallimore, J. M.</dc:creator>
<dc:creator>Parungao, K. A.</dc:creator>
<dc:creator>Fong, J.</dc:creator>
<dc:creator>Suzuki, B. M.</dc:creator>
<dc:creator>Jiang, Z.</dc:creator>
<dc:creator>Isoe, J.</dc:creator>
<dc:creator>Rascon, A. A.</dc:creator>
<dc:date>2024-09-24</dc:date>
<dc:identifier>doi:10.1101/2024.09.23.614410</dc:identifier>
<dc:title><![CDATA[Comprehensive proteolytic profiling of Aedes aegypti mosquito midgut extracts: Unraveling the blood meal protein digestion system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.23.613776v1?rss=1">
<title>
<![CDATA[
Macrophages redeploy functional cancer cell surface proteins following phagocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.23.613776v1?rss=1</link>
<description><![CDATA[
Macrophage-mediated phagocytosis is a vital innate immune process altered in cancer. We show here that tumor-associated macrophages (TAMs) redeploy intact cell surface proteins from cancer cells to their own cell surface. We initially observed the canonical epithelial cancer surface marker EpCAM on the surface of TAMs in primary human solid tumors but not paired peripheral blood macrophages. In a murine model of metastatic breast cancer, we also observed EpCAM on the surface of primary TAMs that have phagocytosed breast cancer cells. In a model of a myeloproliferative neoplasm, we again found engulfed cell-derived surface proteins on the surface of macrophages following phagocytosis. A co-culture system and proteomics assay that tags proteins based on their cell-of-origin revealed hundreds of cell surface proteins synthesized in cancer cells are present and fully intact on the surface of macrophages following phagocytosis. Using a biotin transfer assay, we determined that these proteins were on the surface of the cancer cell prior to redeployment by the macrophage following phagocytosis. Furthermore, macrophages that redeploy a neutral amino acid transporter correspondingly show increased transport of an unnatural amino acid substrate. Widespread acquisition of proteins from engulfed cells may contribute to two critical TAM phenotypes: the inability to phagocytose and reprogrammed metabolism.
]]></description>
<dc:creator>Volk, R. F.</dc:creator>
<dc:creator>Casebeer, S. W.</dc:creator>
<dc:creator>Condon, A. C.</dc:creator>
<dc:creator>Zirak, B.</dc:creator>
<dc:creator>Manon, N.</dc:creator>
<dc:creator>Irkliyenko, I.</dc:creator>
<dc:creator>Liao, H.</dc:creator>
<dc:creator>Tao, S.</dc:creator>
<dc:creator>Pollini, T.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Maker, A.</dc:creator>
<dc:creator>Fidler, T.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Zaro, B. W.</dc:creator>
<dc:date>2024-09-25</dc:date>
<dc:identifier>doi:10.1101/2024.09.23.613776</dc:identifier>
<dc:title><![CDATA[Macrophages redeploy functional cancer cell surface proteins following phagocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.23.614494v1?rss=1">
<title>
<![CDATA[
Histone demethylase enzymes KDM5A and KDM5B modulate immune response by suppressing transcription of endogenous retroviral elements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.23.614494v1?rss=1</link>
<description><![CDATA[
Epigenetic factors, including lysine-specific demethylases such as the KDM5 paralogs KDM5A and KDM5B have been implicated in cancer and the regulation of immune responses. Here, we performed a comprehensive multiomic study in cells lacking KDM5A or KDM5B to map changes in transcriptional regulation and chromatin organization. RNA-seq analysis revealed a significant decrease in the expression of Kruppel-associated box containing zinc finger (KRAB-ZNF) genes in KDM5A or KDM5B knockout cell lines, which was accompanied by changes ATAC-seq and H3K4me3 ChIP-seq. Pharmacological inhibition of KDM5A and KDM5B catalytic activity with a pan-KDM5 inhibitor, CPI-455, did not significantly change KRAB-ZNF expression, raising the possibility that regulation of KRAB-ZNF expression does not require KDM5A and KDM5B demethylase activity. KRAB-ZNF are recognized suppressors of the transcription of endogenous retroviruses (ERVs) and HAP1 cells with KDM5A or KDM5B gene inactivation showed elevated ERV expression, increased dsRNA levels and elevated levels of immune response genes. Acute degradation of KDM5A using a dTAG system in HAP1 cells led to increased ERV expression, demonstrating that de-repression of ERV genes occurs rapidly after loss of KDM5A. Co-immunoprecipitation of KDM5A revealed an interaction with the Nucleosome Remodeling and Deacetylase (NuRD) complex suggesting that KDM5A and NuRD may act together to regulate the expression of ERVs through KRAB-ZNFs. These findings reveal roles of KDM5A and KDM5B in modulating ERV expression and underscore the therapeutic potential of using degraders of KDM5A and KDM5B to modulate tumor immune responses.

Author SummaryThe histone demethylases KDM5A and KDM5B are transcriptional repressors that play an important role in cancer and immune response, making them attractive drug targets. Unfortunately, small molecule inhibitors, including CPI-455, that block KDM5A and KDM5B enzymatic activity, have shown only limited effectiveness at suppressing cancer cell viability as single agents in vitro. In this study we undertook a multi-omics approach to map transcriptional and chromatin changes in KDM5A and KDM5B deficient cells compared to those treated with CPI-455. The datasets revealed that KDM5A and KDM5B modulate the expression of KRAB-ZNF genes and that loss of either gene was associated with increased expression of ERV genes and upregulation of immune response markers. Surprisingly, pharmacological inhibition of these enzymes did not phenocopy genetic ablation. In contrast, acute degradation of KDM5A using a dTAG system caused an increase in ERV expression, providing evidence that this immune modulation is independent of demethylase activity. Together with the limited success of small molecule inhibitors, our data provide strong rationale for the development of KDM5A and KDM5B degraders to modulate tumor immune responses.
]]></description>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Sarah, L.</dc:creator>
<dc:creator>Pucciarelli, D.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Diolaiti, M. E.</dc:creator>
<dc:creator>Galonic Fujimori, D.</dc:creator>
<dc:creator>ashworth, A.</dc:creator>
<dc:date>2024-09-25</dc:date>
<dc:identifier>doi:10.1101/2024.09.23.614494</dc:identifier>
<dc:title><![CDATA[Histone demethylase enzymes KDM5A and KDM5B modulate immune response by suppressing transcription of endogenous retroviral elements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.23.614610v1?rss=1">
<title>
<![CDATA[
nSMase2-mediated exosome secretion shapes the tumor microenvironment to immunologically support pancreatic cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.23.614610v1?rss=1</link>
<description><![CDATA[
The pleiotropic roles of nSMase2-generated ceramide include regulation of intracellular ceramide signaling and exosome biogenesis. We investigated the effects of eliminating nSMase2 on early and advanced PDA, including its influence on the microenvironment. Employing the KPC mouse model of pancreatic cancer, we demonstrate that pancreatic epithelial nSMase2 ablation reduces neoplasia and promotes a PDA subtype switch from aggressive basal-like to classical. nSMase2 elimination prolongs survival of KPC mice, hinders vasculature development, and fosters a robust immune response. nSMase2 loss leads to recruitment of cytotoxic T cells, N1-like neutrophils, and abundant infiltration of anti-tumorigenic macrophages in the pancreatic preneoplastic microenvironment. Mechanistically, we demonstrate that nSMase2-expressing PDA cell small extracellular vesicles (sEVs) reduce survival of KPC mice; PDA cell sEVs generated independently of nSMase2 prolong survival of KPC mice and reprogram macrophages to a proinflammatory phenotype. Collectively, our study highlights previously unappreciated opposing roles for exosomes, based on biogenesis pathway, during PDA progression.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=189 SRC="FIGDIR/small/614610v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Hendley, A. M.</dc:creator>
<dc:creator>Ashe, S.</dc:creator>
<dc:creator>Urano, A.</dc:creator>
<dc:creator>Ng, M.</dc:creator>
<dc:creator>Phu, T. A.</dc:creator>
<dc:creator>Peng, X. L.</dc:creator>
<dc:creator>Luan, C.</dc:creator>
<dc:creator>Finger, A.-M.</dc:creator>
<dc:creator>Jang, G. H.</dc:creator>
<dc:creator>Kerper, N. R.</dc:creator>
<dc:creator>Berrios, D. I.</dc:creator>
<dc:creator>Jin, D.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Riahi, I. R.</dc:creator>
<dc:creator>Gbenedio, O. M.</dc:creator>
<dc:creator>Chung, C.</dc:creator>
<dc:creator>Roose, J.</dc:creator>
<dc:creator>Yeh, J. J.</dc:creator>
<dc:creator>Gallinger, S.</dc:creator>
<dc:creator>Biankin, A. V.</dc:creator>
<dc:creator>OKane, G. M.</dc:creator>
<dc:creator>Ntranos, V.</dc:creator>
<dc:creator>Chang, D.</dc:creator>
<dc:creator>Dawson, D. W.</dc:creator>
<dc:creator>Kim, G. E.</dc:creator>
<dc:creator>Weaver, V. M.</dc:creator>
<dc:creator>Raffai, R. L.</dc:creator>
<dc:creator>Hebrok, M.</dc:creator>
<dc:date>2024-09-25</dc:date>
<dc:identifier>doi:10.1101/2024.09.23.614610</dc:identifier>
<dc:title><![CDATA[nSMase2-mediated exosome secretion shapes the tumor microenvironment to immunologically support pancreatic cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.25.613753v1?rss=1">
<title>
<![CDATA[
Oxytocin receptor controls distinct components of pair bonding and development in prairie voles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.25.613753v1?rss=1</link>
<description><![CDATA[
Oxytocin receptor (Oxtr) signaling influences complex social behaviors in diverse species, including social monogamy in prairie voles. How Oxtr regulates specific components of social attachment behaviors and the neural mechanisms mediating them remains unknown. Here, we examine prairie voles lacking Oxtr and demonstrate that pair bonding comprises distinct behavioral modules: the preference for a bonded partner, and the rejection of novel potential mates. Our longitudinal study of social attachment shows that Oxtr sex-specifically influences early interactions between novel partners facilitating the formation of partner preference. Additionally, Oxtr suppresses promiscuity towards novel potential mates following pair bonding, contributing to rejection. Oxtr function regulates coordinated patterns of gene expression in regions implicated in attachment behaviors and regulates the expression of oxytocin in the paraventricular nucleus of the hypothalamus, a principal source of oxytocin. Thus, Oxtr controls genetically separable components of pair bonding behaviors and coordinates development of the neural substrates of attachment.
]]></description>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Berendzen, K. M.</dc:creator>
<dc:creator>Everitt, A.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Williams, G.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Quine, K.</dc:creator>
<dc:creator>Larios, R. D.</dc:creator>
<dc:creator>Long, K. L. P.</dc:creator>
<dc:creator>Hoglen, N.</dc:creator>
<dc:creator>Sulaman, B. A.</dc:creator>
<dc:creator>Heath, M. C.</dc:creator>
<dc:creator>Sherman, M.</dc:creator>
<dc:creator>Kinkel, M.</dc:creator>
<dc:creator>Cai, A.</dc:creator>
<dc:creator>Galo, D.</dc:creator>
<dc:creator>Caamal, L. C.</dc:creator>
<dc:creator>Goodwin, N. L.</dc:creator>
<dc:creator>Beery, A.</dc:creator>
<dc:creator>Bales, K. L.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Willsey, A. J.</dc:creator>
<dc:creator>Manoli, D. S.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.25.613753</dc:identifier>
<dc:title><![CDATA[Oxytocin receptor controls distinct components of pair bonding and development in prairie voles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.25.614984v1?rss=1">
<title>
<![CDATA[
The human ciliopathy protein RSG1 links the CPLANE complex to transition zone architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.25.614984v1?rss=1</link>
<description><![CDATA[
Cilia are essential organelles and variants in genes governing ciliary function result in ciliopathic diseases. The Ciliogenesis and PLANar polarity Effectors (CPLANE) protein complex is essential for ciliogenesis in animals models but remains poorly defined. Notably, all but one subunit of the CPLANE complex have been implicated in human ciliopathy. Here, we identify three families in which variants in the remaining CPLANE subunit CPLANE2/RSG1 also cause ciliopathy. These patients display cleft palate, tongue lobulations and polydactyly, phenotypes characteristic of Oral-Facial-Digital Syndrome. We further show that these alleles disrupt two vital steps of ciliogenesis, basal body docking and recruitment of intraflagellar transport proteins. Moreover, APMS reveals that Rsg1 binds the CPLANE and also the transition zone protein Fam92 in a GTP-dependent manner. Finally, we show that CPLANE is generally required for normal transition zone architecture. Our work demonstrates that CPLANE2/RSG1 is a causative gene for human ciliopathy and also sheds new light on the mechanisms of ciliary transition zone assembly.
]]></description>
<dc:creator>Vazquez, N.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Valenzuela, I.</dc:creator>
<dc:creator>Cusco, I.</dc:creator>
<dc:creator>Codina, M.</dc:creator>
<dc:creator>Martinez-Gil, N.</dc:creator>
<dc:creator>Valverde, D.</dc:creator>
<dc:creator>Lopez-Solarat, C.</dc:creator>
<dc:creator>Bruel, A.-L.</dc:creator>
<dc:creator>Thauvin-Robinet, C.</dc:creator>
<dc:creator>Steichen, E.</dc:creator>
<dc:creator>Filges, I.</dc:creator>
<dc:creator>Joset, P.</dc:creator>
<dc:creator>De Geyter, J.</dc:creator>
<dc:creator>Mooney, N.</dc:creator>
<dc:creator>Gardner, T.</dc:creator>
<dc:creator>Vaidyanathan, K.</dc:creator>
<dc:creator>Marcotte, E.</dc:creator>
<dc:creator>Roberson, E.</dc:creator>
<dc:creator>Phan, T.</dc:creator>
<dc:creator>Derderian, C.</dc:creator>
<dc:creator>Toriyama, M.</dc:creator>
<dc:creator>Demeter, J.</dc:creator>
<dc:creator>Chavez, M.</dc:creator>
<dc:creator>Aziz-Zanjani, M.</dc:creator>
<dc:creator>Jackson, P.</dc:creator>
<dc:creator>Reiter, J.</dc:creator>
<dc:creator>Tizzano, E.</dc:creator>
<dc:creator>Wallingford, J. B.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.25.614984</dc:identifier>
<dc:title><![CDATA[The human ciliopathy protein RSG1 links the CPLANE complex to transition zone architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.25.614804v1?rss=1">
<title>
<![CDATA[
Effect of chronic upregulation of endocannabinoid signaling in vivo with JZL184 on striatal synaptic plasticity and motor learning in YAC128 Huntington disease mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.25.614804v1?rss=1</link>
<description><![CDATA[
Synaptic dysfunction underlies early sensorimotor and cognitive deficits, and precedes neurodegeneration in a variety of disorders, including Alzheimer, Parkinson and Huntington disease (HD). A monogenic inherited disorder, HD manifests with cognitive, motor and mood disorders associated with progressive degeneration of striatal spiny projection neurons and cortical pyramidal neurons. Cortico-basal ganglia-thalamic loops regulate movement selection and motor learning, which are impaired early in HD. Skilled motor learning is mediated in part by plasticity at cortico-striatal synapses, including endocannabinoid-mediated, high-frequency stimulation induced long-term depression (HFS-LTD). Previously, we found impaired HFS-LTD in brain slice recordings from pre-manifest HD mouse models, which was corrected by JZL184, an inhibitor of endocannabinoid 2-arachidonoyl glycerol (2-AG) degradation. Here, we tested the effects of JZL184 administered in vivo to YAC128 HD model and wild-type (WT) littermate mice. JZL184, given orally daily over a 3-week period, significantly increased levels of 2-AG in striatal tissue. While JZL184 treatment had no impact on open field behavior which was similar for the two genotypes, the treatment improved motor learning on the rotarod task in YAC128 mice to the level observed in WT mice. Moreover, HFS-induced striatal plasticity measured by field potential recording in acute brain slice from YAC128 mice was normalized to WT levels after JZL184 treatment. These results suggest a novel target for mitigating early symptoms of HD, and support the need for clinical trials to test the efficacy of modulating the endocannabinoid system in treatment of HD.
]]></description>
<dc:creator>Sepers, M. D.</dc:creator>
<dc:creator>Woodard, C. L.</dc:creator>
<dc:creator>Ramandi, D.</dc:creator>
<dc:creator>Vecchiarelli, H. A.</dc:creator>
<dc:creator>Hill, M. N.</dc:creator>
<dc:creator>Raymond, L. A.</dc:creator>
<dc:date>2024-09-27</dc:date>
<dc:identifier>doi:10.1101/2024.09.25.614804</dc:identifier>
<dc:title><![CDATA[Effect of chronic upregulation of endocannabinoid signaling in vivo with JZL184 on striatal synaptic plasticity and motor learning in YAC128 Huntington disease mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.25.615027v1?rss=1">
<title>
<![CDATA[
Robust lipid scrambling by TMEM16 proteins requires an open groove and thin membrane 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.25.615027v1?rss=1</link>
<description><![CDATA[
Biological membranes are complex and dynamic structures with different populations of lipids in their inner and outer leaflets. The Ca2+-activated TMEM16 family of membrane proteins plays an important role in collapsing this asymmetric lipid distribution by spontaneously, and bidirectionally, scrambling phospholipids between the two leaflets, which can initiate signaling and alter the physical properties of the membrane. While evidence shows that lipid scrambling can occur via an open hydrophilic pathway ("groove") that spans the membrane, it remains unclear if all family members facilitate lipid movement in this manner. Here we present a comprehensive computational study of lipid scrambling by all TMEM16 members with experimentally solved structures. We performed coarse-grained molecular dynamics (MD) simulations of 27 structures from five different family members solved under activating and non-activating conditions, and we captured over 700 scrambling events in aggregate. This enabled us to directly compare scrambling rates, mechanisms, and protein-lipid interactions for fungal and mammalian TMEM16s, in both open (Ca2+-bound) and closed (Ca2+-free) conformations with statistical rigor. We show that all TMEM16 structures thin the membrane and that the majority of scrambling (>90%) occurs at the groove only when TM4 and TM6 have sufficiently separated. Surprisingly, we also observed 60 scrambling events that occurred outside the canonical groove, over 90% of which took place at the dimer-dimer interface in mammalian TMEM16s. This new site suggests an alternative mechanism for lipid scrambling in the absence of an open groove.

Impact StatementThe majority of TMEM16 lipid scrambling occurs in the open groove associated with Ca2+-activation, but limited scrambling also occurs in the dimer interface independent of Ca2+.
]]></description>
<dc:creator>Stephens, C. A.</dc:creator>
<dc:creator>van Hilten, N.</dc:creator>
<dc:creator>Zheng, L.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:date>2024-09-27</dc:date>
<dc:identifier>doi:10.1101/2024.09.25.615027</dc:identifier>
<dc:title><![CDATA[Robust lipid scrambling by TMEM16 proteins requires an open groove and thin membrane]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.26.615271v1?rss=1">
<title>
<![CDATA[
Dysregulation of the fluid homeostasis system by aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.26.615271v1?rss=1</link>
<description><![CDATA[
Chronic dehydration is a leading cause of morbidity for the elderly, but how aging alters the fluid homeostasis system is not well understood. Here, we used a combination of physiologic, behavioral and circuit analyses to characterize how fluid balance is affected by aging in mice. We found that old mice have a primary defect in sensing and producing the anti-diuretic hormone vasopressin, which results in chronic dehydration. Recordings and manipulations of the thirst circuitry revealed that old mice retain the ability to sense systemic cues of dehydration but are impaired in detecting presystemic, likely oropharyngeal, cues generated during eating and drinking, resulting in disorganized drinking behavior on short timescales. Surprisingly, old mice had increased drinking and motivation after 24-hour water deprivation, indicating that aging does not result in a general impairment in the thirst circuit. These findings reveal how a homeostatic system undergoes coordinated changes during aging.
]]></description>
<dc:creator>Jang, H.</dc:creator>
<dc:creator>Behne Sharma, A.</dc:creator>
<dc:creator>Dan, U.</dc:creator>
<dc:creator>Wong, J. H.</dc:creator>
<dc:creator>Knight, Z. A.</dc:creator>
<dc:creator>Garrison, J. L.</dc:creator>
<dc:date>2024-09-27</dc:date>
<dc:identifier>doi:10.1101/2024.09.26.615271</dc:identifier>
<dc:title><![CDATA[Dysregulation of the fluid homeostasis system by aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.26.615226v1?rss=1">
<title>
<![CDATA[
Tumor Cell Spatial Organization Directs EGFR/RAS/RAF Pathway Primary Therapy Resistance through YAP Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.26.615226v1?rss=1</link>
<description><![CDATA[
Non-small cell lung cancers (NSCLC) harboring common mutations in EGFR and KRAS characteristically respond transiently to targeted therapies against those mutations, but invariably, tumors recur and progress. Resistance often emerges through mutations in the therapeutic target or activation of alternative signaling pathways. Mechanisms of acute tumor cell resistance to initial EGFR (EGFRi) or KRASG12C (G12Ci) pathway inhibition remain poorly understood. Our study reveals that acute response to EGFR/RAS/RAF-pathway inhibition is spatial and culture context specific. In vivo, EGFR mutant tumor xenografts shrink by > 90% following acute EGFRi therapy, and residual tumor cells are associated with dense stroma and have increased nuclear YAP. Interestingly, in vitro EGFRi induced cell cycle arrest in NSCLC cells grown in monolayer, while 3D spheroids preferentially die upon inhibitor treatment. We find differential YAP nuclear localization and activity, driven by the distinct culture conditions, as a common resistance mechanism for selective EGFR/KRAS/BRAF pathway therapies. Forced expression of the YAPS127A mutant partially protects cells from EGFR-mediated cell death in spheroid culture. These studies identify YAP activation in monolayer culture as a non-genetic mechanism of acute EGFR/KRAS/BRAF therapy resistance, highlighting that monolayer vs spheroid cell culture systems can model distinct stages of patient cancer progression.

Graphical Abstract

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]]></description>
<dc:creator>Nakagawa, R.</dc:creator>
<dc:creator>Beardsley, A.</dc:creator>
<dc:creator>Durney, S.</dc:creator>
<dc:creator>Hayward, M.-K.</dc:creator>
<dc:creator>Subramanyam, V.</dc:creator>
<dc:creator>Meyer, N. P.</dc:creator>
<dc:creator>Wismer, H.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Weaver, V. M.</dc:creator>
<dc:creator>Van de Mark, D.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:date>2024-09-28</dc:date>
<dc:identifier>doi:10.1101/2024.09.26.615226</dc:identifier>
<dc:title><![CDATA[Tumor Cell Spatial Organization Directs EGFR/RAS/RAF Pathway Primary Therapy Resistance through YAP Signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.27.615276v1?rss=1">
<title>
<![CDATA[
Selection promotes age-dependent degeneration of the mitochondrial genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.27.615276v1?rss=1</link>
<description><![CDATA[
Somatic mutations in mitochondrial genomes (mtDNA) accumulate exponentially during aging. Using single cell sequencing, we characterize the spectrum of age-accumulated mtDNA mutations in mouse and human liver and identify directional forces that accelerate the accumulation of mutations beyond the rate predicted by a neutral model. "Driver" mutations that give genomes a replicative advantage rose to high cellular abundance and carried along "passenger" mutations, some of which are deleterious. In addition, alleles that alter mtDNA-encoded proteins selectively increased in abundance overtime, strongly supporting the idea of a "destructive" selection that favors genomes lacking function. Overall, this combination of selective forces acting in hepatocytes promotes somatic accumulation of mutations in coding regions of mtDNA that are otherwise conserved in evolution. We propose that these selective processes could contribute to the population prevalence of mtDNA mutations, accelerate the course of heteroplasmic mitochondrial diseases and promote age-associated erosion of the mitochondrial genome.
]]></description>
<dc:creator>Korotkevich, E.</dc:creator>
<dc:creator>Conrad, D. N.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:creator>O'Farrell, P. H.</dc:creator>
<dc:date>2024-09-28</dc:date>
<dc:identifier>doi:10.1101/2024.09.27.615276</dc:identifier>
<dc:title><![CDATA[Selection promotes age-dependent degeneration of the mitochondrial genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.27.615443v1?rss=1">
<title>
<![CDATA[
Latent-TGF-β has a domain swapped architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.27.615443v1?rss=1</link>
<description><![CDATA[
The multifunctional cytokine TGF-{beta} is produced in a latent form (L-TGF-{beta}) where a RGD containing homodimeric prodomain forms a "ring" encircling mature TGF-{beta} shielding it from its receptors. Thus L-TGF-{beta} must be activated to function, a process driven by dynamic allostery resulting from integrin binding the L-TGF-{beta} RGD motif. Here we provide critical evidence that defines a domain-swapped architecture of L-TGF-{beta}, an essential component in the dynamic allostery mechanism of L-TGF-{beta} activation.
]]></description>
<dc:creator>Jin, M.</dc:creator>
<dc:creator>Seed, R.</dc:creator>
<dc:creator>Shing, T.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Nishimura, S.</dc:creator>
<dc:date>2024-09-30</dc:date>
<dc:identifier>doi:10.1101/2024.09.27.615443</dc:identifier>
<dc:title><![CDATA[Latent-TGF-β has a domain swapped architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.28.615614v1?rss=1">
<title>
<![CDATA[
Engineered Feedback Employing Natural Hypoxia-Responsive Factors Enhances Synthetic Hypoxia Biosensors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.28.615614v1?rss=1</link>
<description><![CDATA[
DNA-based hypoxia biosensors conditionally express a gene of interest when a cell is in a state of inadequate oxygen supply, which is a feature of several acute and chronic diseases. These biosensors can be deployed in engineered cells to study or treat disease. Although the central mediators of hypoxia responsiveness have been characterized, the dynamics of this response are generally less understood, and there is no general approach to modulate hypoxia biosensors to tune their performance to meet application-specific needs. To address the need for high-performing hypoxia biosensors, we investigated strategies to enhance biosensor performance by identifying minimal promoter choices and positive feedback circuits that both achieved low background and amplified hypoxia-induced gene expression. To generate insight into the mechanisms by which feedback drives differential performance, we developed an explanatory mathematical model. Our analysis suggests a previously unreported dual regulatory mechanism that was necessary to explain the full set of experimental observations and that provides new insights into regulatory dynamics in chronic hypoxia. This study exemplifies the potential of using synthetic gene circuits to perturb natural systems in a manner that uniquely enables the elucidation of novel facets of natural regulation.
]]></description>
<dc:creator>Dreyer, K. S.</dc:creator>
<dc:creator>Donahue, P. S.</dc:creator>
<dc:creator>Boucher, J. D.</dc:creator>
<dc:creator>Chambers, K. M.</dc:creator>
<dc:creator>Ornelas, M. Y.</dc:creator>
<dc:creator>Edelstein, H. I.</dc:creator>
<dc:creator>Leibowitz, B. D.</dc:creator>
<dc:creator>Zhu, K. J.</dc:creator>
<dc:creator>Dray, K. E.</dc:creator>
<dc:creator>Muldoon, J. J.</dc:creator>
<dc:creator>Leonard, J. N.</dc:creator>
<dc:date>2024-09-30</dc:date>
<dc:identifier>doi:10.1101/2024.09.28.615614</dc:identifier>
<dc:title><![CDATA[Engineered Feedback Employing Natural Hypoxia-Responsive Factors Enhances Synthetic Hypoxia Biosensors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.30.615241v1?rss=1">
<title>
<![CDATA[
Maintenance of neuronal TDP-43 expression requires axonal lysosome transport 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.30.615241v1?rss=1</link>
<description><![CDATA[
TDP-43 mislocalization and pathology occurs across a range of neurodegenerative diseases, but the pathways that modulate TDP-43 in neurons are not well understood. We generated a Halo-TDP-43 knock-in iPSC line and performed a genome-wide CRISPR interference FACS-based screen to identify modifiers of TDP-43 levels in neurons. A meta-analysis of our screen and publicly available screens identified both specific hits and pathways present across multiple screens, the latter likely responsible for generic protein level maintenance. We identified BORC, a complex required for anterograde lysosome transport, as a specific modifier of TDP-43 protein, but not mRNA, levels in neurons. BORC loss led to longer half-life of TDP-43 and other proteins, suggesting lysosome location is required for proper protein turnover. As such, lysosome location and function are crucial for maintaining TDP-43 protein levels in neurons.
]]></description>
<dc:creator>Ryan, V. H.</dc:creator>
<dc:creator>Lawton, S.</dc:creator>
<dc:creator>Reyes, J. F.</dc:creator>
<dc:creator>Hawrot, J.</dc:creator>
<dc:creator>Frankenfield, A. M.</dc:creator>
<dc:creator>Seddighi, S.</dc:creator>
<dc:creator>Ramos, D. M.</dc:creator>
<dc:creator>Johnson, N. L.</dc:creator>
<dc:creator>Zou, J.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Replogle, J.</dc:creator>
<dc:creator>Yuan, H.</dc:creator>
<dc:creator>Johnson, K. R.</dc:creator>
<dc:creator>Maric, D.</dc:creator>
<dc:creator>Hao, L.</dc:creator>
<dc:creator>Nalls, M. A.</dc:creator>
<dc:creator>Ward, M.</dc:creator>
<dc:date>2024-10-01</dc:date>
<dc:identifier>doi:10.1101/2024.09.30.615241</dc:identifier>
<dc:title><![CDATA[Maintenance of neuronal TDP-43 expression requires axonal lysosome transport]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.30.615861v1?rss=1">
<title>
<![CDATA[
Circulatory proteins shape microglia state and boost phagocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.30.615861v1?rss=1</link>
<description><![CDATA[
Microglia, the brains immune cells, are highly responsive to their local environment. Given that circulatory proteins can enter the brain, we asked whether microglia are responsive to such proteins. Here, we identify a stable population of microglia specialized to take up circulatory proteins in a region-specific manner under physiological conditions; human hematopoietic stem cell-derived microglia replacing endogenous microglia in chimeric mice show similar regional specialization. Plasma-positive microglia are characterized by prominent expression of genes related to innate immunity and antigen presentation and exhibit high metabolic and phagocytic activity. This activity is dependent, in part, on microglial uptake and accumulation of circulatory Apolipoprotein AI (ApoA-I). Our findings thus identify a new model of communication between brain and periphery through specialized microglia.
]]></description>
<dc:creator>Lu, N.</dc:creator>
<dc:creator>Moran-Losada, P.</dc:creator>
<dc:creator>Hahn, O.</dc:creator>
<dc:creator>Saksena, A.</dc:creator>
<dc:creator>Tapp, E.</dc:creator>
<dc:creator>Chadarevian, J. P.</dc:creator>
<dc:creator>Dong, W.</dc:creator>
<dc:creator>Shi, S. M.</dc:creator>
<dc:creator>Shuken, S. R.</dc:creator>
<dc:creator>Guldner, I.</dc:creator>
<dc:creator>Zeng, W.</dc:creator>
<dc:creator>To, N.-S.</dc:creator>
<dc:creator>Wong, P. S.</dc:creator>
<dc:creator>Hasselmann, J.</dc:creator>
<dc:creator>Davtyan, H.</dc:creator>
<dc:creator>Sun, J.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Yang, A. C.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Cheung, T. H.</dc:creator>
<dc:creator>Abu-Remaileh, M.</dc:creator>
<dc:creator>Blurton-Jones, M.</dc:creator>
<dc:creator>Wyss-Coray, T.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.09.30.615861</dc:identifier>
<dc:title><![CDATA[Circulatory proteins shape microglia state and boost phagocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.30.615930v1?rss=1">
<title>
<![CDATA[
Robust Detection of Brain Stimulation Artifacts in iEEG Using Autoencoder-Generated Signals and ResNet Classification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.30.615930v1?rss=1</link>
<description><![CDATA[
BackgroundIntracranial EEG (iEEG) is crucial for understanding brain function, but stimulation-induced noise complicates data interpretation. Traditional artifact detection methods require manual user input or struggle with noise variability, especially with limited labeled data.

ObjectiveWe developed a supervised method to automatically detect stimulation-induced noise in human iEEG recordings using synthetic data generated by Variational Autoencoders (VAEs) to train a ResNet-18 classifier.

MethodsMulti-lead iEEG data were collected, preprocessed, and used to train VAEs for generating synthetic clean and noisy signals. The ResNet-18 model was trained on images of spectra generated from these synthetic signals and validated on real iEEG data from five participants.

ResultsThe classifier, trained exclusively on synthetic data, demonstrated high accuracy, precision, and recall when applied to real iEEG recordings, with AUC values greater than 0.99 across all participants.

ConclusionWe present a novel approach to effectively detect stimulation-induced noise in iEEG, offering a robust solution for improving data interpretation in scenarios with limited labeled data. Additionally, the pre-trained ResNet-18 model is available for the community to use, facilitating further research and application in similar datasets.
]]></description>
<dc:creator>Saal, J.</dc:creator>
<dc:creator>Khambhati, A. N.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Shirvalkar, P.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.09.30.615930</dc:identifier>
<dc:title><![CDATA[Robust Detection of Brain Stimulation Artifacts in iEEG Using Autoencoder-Generated Signals and ResNet Classification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.01.616097v1?rss=1">
<title>
<![CDATA[
Interrogating the plasma proteome of repetitive head impact exposure and chronic traumatic encephalopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.01.616097v1?rss=1</link>
<description><![CDATA[
BackgroundExposure to repetitive head impacts (RHI) is associated with increased risk for chronic traumatic encephalopathy (CTE), a neurodegenerative tauopathy, and other neuropathological changes. Biological drivers of RHI-related neurodegeneration are not well understood. We interrogated the plasma proteome in aging adults with prior RHI compared to healthy controls (CTL) and individuals with Alzheimers disease (AD), including a subset characterized neuropathologically at autopsy.

MethodsProximity extension assay (Olink Explore(R)) quantified 2,779 plasma proteins in 22 RHI patients (all AD-biomarker negative), 39 biomarker-confirmed AD, and 44 CTL. A subset of participants went to autopsy (N=16) allowing for comparisons of the antemortem plasma proteome between autopsy-confirmed CTE+ (N=7) and CTE-(N=9). Differential abundance and co-expression network analyses identified plasma proteomic signatures of RHI, which were functionally annotated using gene ontology and cell type enrichment analysis. Nonparametric correlations examined plasma proteomic associations with orthogonally-measured plasma biomarkers, global cognitive function, and semi-quantitative ratings of neuropathology burden at autopsy.

ResultsDifferential abundance analysis revealed 434 increased (vs. 6 decreased) proteins in RHI vs. CTL and 193 increased (vs. 14 decreased) in RHI vs. AD. Network analysis identified 9 protein co-expression modules (M1-M9), of which 7 were elevated in RHI compared to AD or CTL. Modules with increased abundance in RHI were enriched for mitochondrial/metabolic, cell division, and immunovascular (e.g., cell adhesion, TNF-signaling) processes. RHI-related modules exhibited strong and selective correlations with immunoassay-based plasma IL-6 in RHI cases, including the M2 TNF-signaling/cell adhesion module which harbored proteins that strongly tracked with cognitive function. RHI-related plasma protein signatures were similar in the subset of participants with autopsy-confirmed CTE, including immune and metabolic modules that positively correlated with medial temporal lobe tau and TDP-43 burden.

ConclusionsMolecular pathways in plasma most consistently implicated in RHI were tied to immune response, mitochondrial function, and cell metabolism. RHI-related proteomic signatures tracked with antemortem cognitive severity and postmortem neuropathological burden, providing converging evidence for their role in disease progression. Differentially abundant proteins and co-expression modules in RHI may inform mechanisms linking RHI to increased dementia risk, thus guiding diagnostic biomarker and therapeutic development for at-risk populations.
]]></description>
<dc:creator>Saloner, R.</dc:creator>
<dc:creator>Casaletto, K. B.</dc:creator>
<dc:creator>Rayaprolu, S.</dc:creator>
<dc:creator>Chakrabarty, P.</dc:creator>
<dc:creator>Abisambra, J. F.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Kramer, J. H.</dc:creator>
<dc:creator>Rabinovici, G. D.</dc:creator>
<dc:creator>Asken, B. M.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.616097</dc:identifier>
<dc:title><![CDATA[Interrogating the plasma proteome of repetitive head impact exposure and chronic traumatic encephalopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.01.616161v1?rss=1">
<title>
<![CDATA[
Long range mutual activation establishes Rho and Rac polarity during cell migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.01.616161v1?rss=1</link>
<description><![CDATA[
In migrating cells, the GTPase Rac organizes a protrusive front, whereas Rho organizes a contractile back. How these GTPases are appropriately positioned at the opposite poles of migrating cells is unknown. Here we leverage optogenetics, manipulation of cell mechanics, and mathematical modeling to reveal a surprising mechanochemical long-range mutual activation of the front and back polarity programs that complements their well-known local mutual inhibition. Rac-based protrusion stimulates Rho activation at the opposite side of the cell via membrane tension-based activation of mTORC2. Conversely, Rho-based contraction induces cortical-flow-based regulation of phosphoinositide signaling to trigger Rac activation at the opposite side of the cell. We develop a minimal unifying mechanochemical model of the cell to explain how this long-range facilitation complements local inhibition to enable robust Rho and Rac partitioning. We show that this long-range mutual activation of Rac and Rho is conserved in epithelial cells and is also essential for efficient polarity and migration of primary human T cells, indicating the generality of this circuit. Our findings demonstrate that the actin cortex and plasma membrane function as an integrated mechanochemical system for long-range partitioning of Rac and Rho during cell migration and likely other cellular contexts.
]]></description>
<dc:creator>De Belly, H.</dc:creator>
<dc:creator>Gallen, A. F.</dc:creator>
<dc:creator>Strickland, E.</dc:creator>
<dc:creator>Estrada, D. C.</dc:creator>
<dc:creator>Zager, P. J.</dc:creator>
<dc:creator>Burkhardt, J. K.</dc:creator>
<dc:creator>Turlier, H.</dc:creator>
<dc:creator>Weiner, O.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.616161</dc:identifier>
<dc:title><![CDATA[Long range mutual activation establishes Rho and Rac polarity during cell migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.01.616167v1?rss=1">
<title>
<![CDATA[
High-Complexity Barcoded Rabies Virus for Scalable Circuit Mapping Using Single-Cell and Single-Nucleus Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.01.616167v1?rss=1</link>
<description><![CDATA[
Single cell genomics has revolutionized our understanding of neuronal cell types. However, scalable technologies for probing single-cell connectivity are lacking, and we are just beginning to understand how molecularly defined cell types are organized into functional circuits. Here, we describe a protocol to generate high-complexity barcoded rabies virus (RV) for scalable circuit mapping from tens of thousands of individual starter cells in parallel. In addition, we introduce a strategy for targeting RV-encoded barcode transcripts to the nucleus so that they can be read out using single-nucleus RNA sequencing (snRNA-seq). We apply this tool in organotypic slice cultures of the developing human cerebral cortex, which reveals the emergence of cell type- specific circuit motifs in midgestation. By leveraging the power and throughput of single cell genomics for mapping synaptic connectivity, we chart a path forward for scalable circuit mapping of molecularly-defined cell types in healthy and disease states.
]]></description>
<dc:creator>Shin, D.</dc:creator>
<dc:creator>Urbanek, M. E.</dc:creator>
<dc:creator>Larson, H. H.</dc:creator>
<dc:creator>Moussa, A. J.</dc:creator>
<dc:creator>Lee, K. Y.</dc:creator>
<dc:creator>Baker, D. L.</dc:creator>
<dc:creator>Standen-Bloom, E.</dc:creator>
<dc:creator>Ramachandran, S.</dc:creator>
<dc:creator>Bogdanoff, D.</dc:creator>
<dc:creator>Cadwell, C. R.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.616167</dc:identifier>
<dc:title><![CDATA[High-Complexity Barcoded Rabies Virus for Scalable Circuit Mapping Using Single-Cell and Single-Nucleus Sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.01.614851v1?rss=1">
<title>
<![CDATA[
Sex differences in the clinical manifestation of autosomal dominant frontotemporal dementia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.01.614851v1?rss=1</link>
<description><![CDATA[
INTRODUCTIONSex differences are apparent in neurodegenerative diseases, but have not been comprehensively characterized in frontotemporal dementia (FTD).

METHODSParticipants included 337 adults with autosomal dominant FTD enrolled in the ALLFTD Consortium. Clinical assessments and plasma were collected annually for up to six years. Linear mixed-effects models investigated how sex and disease stage associated with longitudinal trajectories of cognition, function, and neurofilament light chain (NfL).

RESULTSWhile sex differences were not apparent at asymptomatic stages, females showed more rapid declines across all outcomes in symptomatic stages compared to males. In asymptomatic participants, the association between baseline NfL and clinical trajectories was weaker in females versus males, a difference that attenuated in symptomatic participants.

DISCUSSIONIn genetic FTD, females show cognitive resilience in early disease stages followed by steeper clinical declines later in disease. Baseline NfL may be a less sensitive prognostic tool for clinical progression in females with FTD-causing mutations.
]]></description>
<dc:creator>Memel, M.</dc:creator>
<dc:creator>Staffaroni, A.</dc:creator>
<dc:creator>Ilan-Gala, I.</dc:creator>
<dc:creator>Garcia Castro, J.</dc:creator>
<dc:creator>Kornak, J.</dc:creator>
<dc:creator>Tartaglia, C.</dc:creator>
<dc:creator>Saloner, R.</dc:creator>
<dc:creator>VandeBunte, A.</dc:creator>
<dc:creator>Paolillo, E.</dc:creator>
<dc:creator>Cadwallader, C.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Gorno-Tempini, M. L.</dc:creator>
<dc:creator>Mandelli, M.</dc:creator>
<dc:creator>Apostolova, L.</dc:creator>
<dc:creator>Graff-Radford, N.</dc:creator>
<dc:creator>Litvan, I.</dc:creator>
<dc:creator>Bayram, E.</dc:creator>
<dc:creator>Pressman, P.</dc:creator>
<dc:creator>Miyagawa, T.</dc:creator>
<dc:creator>Mackenzie, I.</dc:creator>
<dc:creator>Goldman, J.</dc:creator>
<dc:creator>Darby, R.</dc:creator>
<dc:creator>Appleby, B.</dc:creator>
<dc:creator>Petrucelli, L.</dc:creator>
<dc:creator>Gendron, T.</dc:creator>
<dc:creator>Heuer, H.</dc:creator>
<dc:creator>Forseberg, L.</dc:creator>
<dc:creator>Rojas, J.</dc:creator>
<dc:creator>Boeve, B.</dc:creator>
<dc:creator>Brushaber, N.</dc:creator>
<dc:creator>Domoto-Reilly, K.</dc:creator>
<dc:creator>Ghoshal, N.</dc:creator>
<dc:creator>Lapid, M.</dc:creator>
<dc:creator>Pascual, B.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Ramos, E.</dc:creator>
<dc:creator>Ramanan, V.</dc:creator>
<dc:creator>Rademakers, R.</dc:creator>
<dc:creator>Rascovsky, K.</dc:creator>
<dc:creator>Pantelyat, A.</dc:creator>
<dc:creator>Masdeu, J.</dc:creator>
<dc:creator>Snyder, A.</dc:creator>
<dc:creator>Boxer, A.</dc:creator>
<dc:creator>Rosen</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.614851</dc:identifier>
<dc:title><![CDATA[Sex differences in the clinical manifestation of autosomal dominant frontotemporal dementia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.01.616181v1?rss=1">
<title>
<![CDATA[
Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.01.616181v1?rss=1</link>
<description><![CDATA[
Dinoflagellates and cyanobacteria in harmful  red tide algal blooms produce saxitoxin (STX) and [~]50 congeners that block voltage-gated sodium channel (NaV) function and disrupt bioelectrical signals1-4. Consuming seafood carrying these lethal toxins causes paralytic shellfish poisoning (PSP), a growing public health hazard due to climate change5-7 that motivates efforts to detect these toxins and counteract their noxious effects. Although structural studies of NaVs8,9 and anuran soluble STX binding proteins known as saxiphilins (Sxphs)10,11 revealed convergent binding modes for the bis-guanidinium STX core10,11, the structural basis for STX congener recognition is unknown. Here, we show that American bullfrog (Rana catesbeiana) RcSxph10,11 and High Himalaya frog (Nanorana parkeri) NpSxph10 use a pre-organized pocket to sequester STX congeners through a binding mode shared with STX. This  lock and key recognition yields a tradeoff between a relatively rigid high-affinity toxin binding site in which bound waters are crucial and the ability of Sxphs to accommodate STX congener modifications. Importantly, functional studies show that Sxphs act as  toxin sponges that reverse NaV block by multiple STX congeners and can detect these bis-guanidinium toxins in a radioligand receptor binding assay (RBA) for PSP toxin environmental testing12,13. Our findings establish how Sxphs sequester diverse neurotoxins and reveal structural factors underlying STX congener binding differences between Sxphs and NaVs that are rooted in the distinct toxin binding orientations on these two targets. These insights expand the molecular foundation required for understanding toxin sponge action and for guiding development of new means to monitor PSTs and mitigate their harmful effects.
]]></description>
<dc:creator>Zakrzewska, S.</dc:creator>
<dc:creator>Nixon, S. A.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Hajare, H. S.</dc:creator>
<dc:creator>Park, E. R.</dc:creator>
<dc:creator>Mulcahy, J. V.</dc:creator>
<dc:creator>Arlinghaus, K. M.</dc:creator>
<dc:creator>Neu, E.</dc:creator>
<dc:creator>Konovalov, K.</dc:creator>
<dc:creator>Provasi, D.</dc:creator>
<dc:creator>Leighfield, T. A.</dc:creator>
<dc:creator>Filizola, M.</dc:creator>
<dc:creator>Du Bois, J.</dc:creator>
<dc:creator>Minor, D. L.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.616181</dc:identifier>
<dc:title><![CDATA[Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.01.616208v1?rss=1">
<title>
<![CDATA[
microbetag: simplifying microbial network interpretation through annotation, enrichment tests and metabolic complementarity analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.01.616208v1?rss=1</link>
<description><![CDATA[
Microbial co-occurrence network inference is often hindered by low accuracy and tool dependency. We introduce microbetag, a comprehensive software ecosystem designed to enhance network annotation. Nodes (taxa) are enriched with phenotypic traits, while edges represent metabolic complementarities, highlighting potential cross-feeding relationships.

microbetags online version relies on microbetagDB, a database of 34,608 high-quality genomes with detailed annotations. A stand-alone tool allows users to apply microbetag to custom reference genomes/bins/MAGs. Additionally, MGG, a CytoscapeApp, offers a streamlined, user-friendly interface for network retrieval and visualization. microbetag effectively identified known metabolic interactions and serves as a robust hypothesis-generating tool.
]]></description>
<dc:creator>Zafeiropoulos, H.</dc:creator>
<dc:creator>Michail Delopoulos, E. I.</dc:creator>
<dc:creator>Erega, A.</dc:creator>
<dc:creator>Schneider, A.</dc:creator>
<dc:creator>Geirnaert, A.</dc:creator>
<dc:creator>Morris, J.</dc:creator>
<dc:creator>Faust, K.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.01.616208</dc:identifier>
<dc:title><![CDATA[microbetag: simplifying microbial network interpretation through annotation, enrichment tests and metabolic complementarity analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.02.616243v1?rss=1">
<title>
<![CDATA[
Short-term caloric restriction or resveratrol supplementation alters large-scale brain network connectivity in male and female rats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.02.616243v1?rss=1</link>
<description><![CDATA[
IntroductionDietary interventions such as caloric restriction (CR) exert positive effects on brain health. Unfortunately, poor compliance hinders the success of this approach. A proposed alternative is resveratrol (Rsv), a CR-mimetic known to promote brain health. Direct comparison between the effects of Rsv and CR on brain health is lacking, with limited knowledge on their sex-specific effects. Therefore, we aimed to compare and unravel the sex-specific impact of these dietary interventions on spontaneous brain activity.

MethodsHere, we used resting-state fMRI to investigate functional connectivity (FC) changes in five prominent resting-state brain networks (RSNs) in healthy four month old male and female F344 rats supplemented to either 40% CR or daily Rsv supplementation (10 mg/kg, oral) for the duration of one month.

ResultsOur results demonstrated a decreased body weight (BW) in CR rats, as well as an increase in body weight in male Rsv supplemented rats, compared to female Rsv supplemented rats, whereas this difference between sexes was not observed in the control or CR groups. Furthermore, we found that both CR or Rsv supplementation induce a female-specific decrease of FC between the subcortical network and hippocampal network, and between the subcortical network and lateral cortical network. Moreover, Rsv supplementation lowered FC within the hippocampal network and between the hippocampal and the default mode like network, the lateral cortical network and the sensory network - an effect not observed for the CR rats. Finally, we observed an overall lower FC in male rats compared to females, irrespective of dietary intervention, within the subcortical network and between the subcortical, the sensory and default mode like network.

DiscussionOur findings reveal that both CR and Rsv induce a similar female-specific decrease of FC in RSNs associated with memory and emotion, all the while CR and Rsv induce dissimilar changes in body weight and other within- and between RSN FC measures. Altogether, this study provides insight into the effects and comparability of short-term CR and Rsv supplementation on brain connectivity within and between RSNs in both male and female F344 rats, providing a FC reference for future research of dietary effects.
]]></description>
<dc:creator>van Rooij, J. R. A.</dc:creator>
<dc:creator>van den Berg, M.</dc:creator>
<dc:creator>Vasilkosvka, T.</dc:creator>
<dc:creator>Van Audekerke, J.</dc:creator>
<dc:creator>Kosten, L.</dc:creator>
<dc:creator>Bertoglio, D.</dc:creator>
<dc:creator>Adhikari, M. H.</dc:creator>
<dc:creator>Verhoye, M.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.02.616243</dc:identifier>
<dc:title><![CDATA[Short-term caloric restriction or resveratrol supplementation alters large-scale brain network connectivity in male and female rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.02.616359v1?rss=1">
<title>
<![CDATA[
Coordinate post-transcriptional regulation by microRNAs and RNA binding proteins is critical for early embryonic cell fate decisions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.02.616359v1?rss=1</link>
<description><![CDATA[
Abstract/SummaryPost-transcriptional control by RNA binding proteins (RBPs) and microRNAs play central roles in mRNA stability and translation (1). However, little is known about how RBPs and microRNAs coordinate in developmental time to regulate cell fate. Here, we show that coordinate RBP and microRNA control of a single transcript, Profilin 2 (Pfn2), is essential for differentiation of embryonic stem cells (ESCs) into the primary germ layer lineages. The Pfn2 3untranslated region has a binding site for Iron Regulatory Proteins and a nearby binding site for ESC enriched microRNAs (2,3). Deletion of this microRNA site leads to increased PFN2 and reduced FGF signaling during pluripotency transition prior to germ layer formation (4). In contrast, deletion of the iron response element leads to decreased PFN2 and a Wnt signaling defect, reduced nuclear beta-catenin, and a subsequent block in mesendodermal lineages during early germ layer formation. The choreographed microRNA-IRE axis of control on the Pfn2 transcript is essential for two key signal transduction steps during ESC differentiation.
]]></description>
<dc:creator>Sangokoya, C.</dc:creator>
<dc:creator>Blelloch, R.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.02.616359</dc:identifier>
<dc:title><![CDATA[Coordinate post-transcriptional regulation by microRNAs and RNA binding proteins is critical for early embryonic cell fate decisions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.03.616574v1?rss=1">
<title>
<![CDATA[
SPLASH-structure: a statistical approach to identify RNA secondary structures from raw sequencing data, bypassing multiple sequence alignment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.03.616574v1?rss=1</link>
<description><![CDATA[
RNA secondary and tertiary structures are essential to life. Experimental methods to detect RNA structure, such as X-ray crystallography and chemical probing, are incisive but suffer from low throughput and dimensionality. Computational approaches, leveraging evolutionary signals from correlated mutations, provide an alternative means to infer RNA structures. However, these methods require assembly and face challenges due to statistical biases inherent in multiple sequence alignment (MSA). Furthermore, these methods cannot exploit a given RNA elements full spectrum of natural sequence variations. Here, we introduce STRUCT (Statistical Testing of RNA Units with Covariation Traits), an assembly-free, MSA-free, and metadata-free statistical method for identifying conserved RNA structures from raw sequencing data, quantifying base-pair covariations or stem variation exclusion in the putative RNA structures. We show STRUCT rediscovers known HIV structural elements and identifies conserved rRNA structures in metatranscriptomics samples. Moreover, STRUCT finds viral structures in mosquito metatranscriptomics samples de novo, including previously unannotated viral genomes, highlighting the methods potential for viral discovery. STRUCT is an ultra-fast, easy-to-use, and robust tool that excels in high-throughput RNA structure prediction and hypothesis generation, presenting a novel approach for discovering structural RNA elements.
]]></description>
<dc:creator>Wang, J. F.</dc:creator>
<dc:creator>Rustagi, A.</dc:creator>
<dc:creator>Salzman, J.</dc:creator>
<dc:date>2024-10-04</dc:date>
<dc:identifier>doi:10.1101/2024.10.03.616574</dc:identifier>
<dc:title><![CDATA[SPLASH-structure: a statistical approach to identify RNA secondary structures from raw sequencing data, bypassing multiple sequence alignment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.04.615095v1?rss=1">
<title>
<![CDATA[
Synergy between cis-regulatory elements can render cohesin dispensable for distal enhancer function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.04.615095v1?rss=1</link>
<description><![CDATA[
Enhancers are critical genetic elements controlling transcription from promoters, but the mechanisms by which they convey regulatory information across large genomic distances remain elusive. Here, we engineered pluripotent stem cells in which cohesin loop extrusion can be inducibly disrupted without causing confounding cell cycle defects. While evident, transcriptional dysregulation was cell-type specific, and not all loci with distal enhancers depend equally on cohesin extrusion. Using comparative genome editing, we demonstrate that enhancer-promoter communication across as little as 20 kilobases can rely on cohesin. However, promoter-proximal regulatory elements can support long-range, cohesin-independent enhancer action - either upon disabling extrusion or across strong CTCF insulators. Finally, transcriptional dynamics and the emergence of new embryonic cell types in response to differentiation cues remained largely robust to disrupting cohesin extrusion. Beyond establishing novel experimental strategies to study cohesin functions in enhancer biology, our work provides mechanistic insight accounting for both cell type- and genomic context-specificity.
]]></description>
<dc:creator>Hansen, K. L.</dc:creator>
<dc:creator>Adachi, A. S.</dc:creator>
<dc:creator>Braccioli, L.</dc:creator>
<dc:creator>Kadvani, S.</dc:creator>
<dc:creator>Boileau, R. M.</dc:creator>
<dc:creator>Pokorny, B.</dc:creator>
<dc:creator>Shah, R.</dc:creator>
<dc:creator>Anderson, E. C.</dc:creator>
<dc:creator>Martinovic, M.</dc:creator>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Carel, I.</dc:creator>
<dc:creator>Bonitto, K.</dc:creator>
<dc:creator>Blelloch, R.</dc:creator>
<dc:creator>Fudenberg, G.</dc:creator>
<dc:creator>de Wit, E.</dc:creator>
<dc:creator>Nora, E. P.</dc:creator>
<dc:date>2024-10-04</dc:date>
<dc:identifier>doi:10.1101/2024.10.04.615095</dc:identifier>
<dc:title><![CDATA[Synergy between cis-regulatory elements can render cohesin dispensable for distal enhancer function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.08.616922v1?rss=1">
<title>
<![CDATA[
CRISPR tiling deletion screens reveal functional enhancers of neuropsychiatric risk genes and allelic compensation effects (ACE) on transcription 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.08.616922v1?rss=1</link>
<description><![CDATA[
Precise transcriptional regulation is critical for cellular function and development, yet the mechanism of this process remains poorly understood for many genes. To gain a deeper understanding of the regulation of neuropsychiatric disease risk genes, we identified a total of 39 functional enhancers for four dosage-sensitive genes, APP, FMR1, MECP2, and SIN3A, using CRISPR tiling deletion screening in human induced pluripotent stem cell (iPSC)-induced excitatory neurons. We found that enhancer annotation provides potential pathological insights into disease-associated copy number variants. More importantly, we discovered that allelic enhancer deletions at SIN3A could be compensated by increased transcriptional activities from the other intact allele. Such allelic compensation effects (ACE) on transcription is stably maintained during differentiation and, once established, cannot be reversed by ectopic SIN3A expression. Further, ACE at SIN3A occurs through dosage sensing by the promoter. Together, our findings unravel a regulatory compensation mechanism that ensures stable and precise transcriptional output for SIN3A, and potentially other dosage-sensitive genes.
]]></description>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Zheng, L.</dc:creator>
<dc:creator>Maliskova, L.</dc:creator>
<dc:creator>Tam, T. W.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Takagi, M. A.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Ren, B.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:date>2024-10-10</dc:date>
<dc:identifier>doi:10.1101/2024.10.08.616922</dc:identifier>
<dc:title><![CDATA[CRISPR tiling deletion screens reveal functional enhancers of neuropsychiatric risk genes and allelic compensation effects (ACE) on transcription]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.08.617294v1?rss=1">
<title>
<![CDATA[
Proteomic profiling of the local and systemic immune response to pediatric respiratory viral infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.08.617294v1?rss=1</link>
<description><![CDATA[
Viral lower respiratory tract infection (vLRTI) is a leading cause of hospitalization and death in children worldwide. Despite this, no studies have employed proteomics to characterize host immune responses to severe pediatric vLRTI in both the lower airway and systemic circulation. To address this gap, gain insights into vLRTI pathophysiology, and test a novel diagnostic approach, we assayed 1,305 proteins in tracheal aspirate (TA) and plasma from 62 critically ill children using SomaScan. We performed differential expression (DE) and pathway analyses comparing vLRTI (n=40) to controls with non-infectious acute respiratory failure (n=22), developed a diagnostic classifier using LASSO regression, and analyzed matched TA and plasma samples. We further investigated the impact of viral load and bacterial coinfection on the proteome. The TA signature of vLRTI was characterized by 200 DE proteins (Padj<0.05) with upregulation of interferons and T cell responses and downregulation of inflammation-modulating proteins including FABP and MIP-5. A nine-protein TA classifier achieved an AUC of 0.96 (95% CI 0.90-1.00) for identifying vLRTI. In plasma, the host response to vLRTI was more muted with 56 DE proteins. Correlation between TA and plasma was limited, although ISG15 was elevated in both compartments. In bacterial coinfection, we observed increases in the TNF-stimulated protein TSG-6, as well as CRP, and interferon-related proteins. Viral load correlated positively with interferon signaling and negatively with neutrophil-activation pathways. Taken together, our study provides fresh insight into the lower airway and systemic proteome of severe pediatric vLRTI, and identifies novel protein biomarkers with diagnostic potential.

IMPORTANCEWe describe the first proteomic profiling of the lower airway and blood in critically ill children with severe viral lower respiratory tract infection (vLRTI). From tracheal aspirate (TA), we defined a proteomic signature of vLRTI characterized by increased expression of interferon signaling proteins and decreased expression of proteins involved in immune modulation including FABP and MIP-5. Using machine learning, we developed a parsimonious diagnostic classifier that distinguished vLRTI from non-infectious respiratory failure with high accuracy. Comparative analysis of paired TA and plasma specimens demonstrated limited concordance, although the interferon-stimulated protein ISG15 was significantly upregulated with vLRTI in both compartments. We further identified TSG-6 and CRP as airway biomarkers of bacterial-viral coinfection, and viral load analyses demonstrated positive correlation with interferon-related protein expression and negative correlation with the expression of neutrophil activation proteins. Taken together, our study provides new insight into the lower airway and systemic proteome of severe pediatric vLRTI.
]]></description>
<dc:creator>Lydon, E.</dc:creator>
<dc:creator>Osborne, C. M.</dc:creator>
<dc:creator>Wagner, B. D.</dc:creator>
<dc:creator>Ambroggio, L.</dc:creator>
<dc:creator>Harris, J. K.</dc:creator>
<dc:creator>Reeder, R. W.</dc:creator>
<dc:creator>Carpenter, T. C.</dc:creator>
<dc:creator>Maddux, A. B.</dc:creator>
<dc:creator>Leroue, M. K.</dc:creator>
<dc:creator>Yehya, N.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Hall, M. W.</dc:creator>
<dc:creator>Zuppa, A.</dc:creator>
<dc:creator>Carcillo, J. A.</dc:creator>
<dc:creator>Meert, K.</dc:creator>
<dc:creator>Sapru, A.</dc:creator>
<dc:creator>Pollack, M. M.</dc:creator>
<dc:creator>McQuillen, P.</dc:creator>
<dc:creator>Notterman, D.</dc:creator>
<dc:creator>Langelier, C. R.</dc:creator>
<dc:creator>Mourani, P. M.</dc:creator>
<dc:date>2024-10-10</dc:date>
<dc:identifier>doi:10.1101/2024.10.08.617294</dc:identifier>
<dc:title><![CDATA[Proteomic profiling of the local and systemic immune response to pediatric respiratory viral infections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.10.617484v1?rss=1">
<title>
<![CDATA[
Heterozygous and generalist MxA super-restrictors overcome breadth-specificity tradeoffs in antiviral restriction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.10.617484v1?rss=1</link>
<description><![CDATA[
Antiviral restriction factors such as MxA (myxovirus resistance protein A) inhibit a broad range of viruses. However, they face the challenge of maintaining this breadth as viruses evolve to escape their defense. Viral escape drives restriction factors to evolve rapidly, selecting for amino acid changes at their virus-binding interfaces to regain defense. How do restriction factors balance the breadth of antiviral functions against the need to evolve specificity against individual escaping viruses? We explored this question in human MxA, which uses its rapidly evolving loop L4 as the specificity determinant for orthomyxoviruses such as THOV and IAV. Previous combinatorial mutagenesis of rapidly evolving residues in human MxA loop L4 revealed variants with a ten-fold increase in potency against THOV. However, this strategy did not yield improved IAV restriction, suggesting a strong tradeoff between antiviral specificity and breadth. Here, using a modified combinatorial mutagenesis strategy, we find  super-restrictor MxA variants with over ten-fold enhanced restriction of the avian IAV strain H5N1 but reduced THOV restriction. Analysis of super-restrictor MxA variants reveals that the identity of residue 561 explains most of MxAs breadth-specificity tradeoff in H5N1 versus THOV restriction. However, rare  generalist super-restrictors with enhanced restriction of both viruses allow MxA to overcome the breadth-specificity tradeoff. Finally, we show that a heterozygous combination of two  specialist super-restrictors, one against THOV and the other against IAV, enhances restriction against both viruses. Thus, two strategies enable restriction factors such as MxA to increase their restriction of diverse viruses to overcome breadth-specificity tradeoffs that may be pervasive in host-virus conflicts.
]]></description>
<dc:creator>Geiger, R. A.</dc:creator>
<dc:creator>Khera, D.</dc:creator>
<dc:creator>Tenthorey, J.</dc:creator>
<dc:creator>Kochs, G.</dc:creator>
<dc:creator>Graf, L.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2024-10-10</dc:date>
<dc:identifier>doi:10.1101/2024.10.10.617484</dc:identifier>
<dc:title><![CDATA[Heterozygous and generalist MxA super-restrictors overcome breadth-specificity tradeoffs in antiviral restriction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.07.617070v1?rss=1">
<title>
<![CDATA[
Antibody-Fab and -Fc features promote Mycobacterium tuberculosis restriction. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.07.617070v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), is a leading cause of death by an infectious disease globally, with no efficacious vaccine. Antibodies are implicated in Mtb control, but the mechanisms of antibody action remain poorly understood. We assembled a library of TB monoclonal antibodies (mAb) and screened for the ability to restrict Mtb in mice, identifying protective antibodies targeting known and novel antigens. To dissect the mechanism of mAb-mediated Mtb restriction, we optimized a protective lipoarabinomannan-specific mAb through Fc-swapping. In vivo analysis of these Fc-variants revealed a critical role for Fc-effector function in Mtb restriction. Restrictive Fc-variants altered distribution of Mtb across innate immune cells. Single-cell transcriptomics highlighted distinctly activated molecular circuitry within innate immune cell subpopulations, highlighting early activation of neutrophils as a key signature of mAb-mediated Mtb restriction. Therefore, improved antibody-mediated restriction of Mtb is associated with reorganization of the tissue-level immune response to infection and depends on the collaboration of antibody Fab and Fc.
]]></description>
<dc:creator>Grace, P. S.</dc:creator>
<dc:creator>Peters, J. M.</dc:creator>
<dc:creator>Sixsmith, J.</dc:creator>
<dc:creator>Lu, R.</dc:creator>
<dc:creator>Fenderson, B. A.</dc:creator>
<dc:creator>Vickers, A.</dc:creator>
<dc:creator>Slein, M. D.</dc:creator>
<dc:creator>Irvine, E. B.</dc:creator>
<dc:creator>McKitrick, T.</dc:creator>
<dc:creator>Wei, M.-H.</dc:creator>
<dc:creator>Cummings, R. D.</dc:creator>
<dc:creator>Wallace, A.</dc:creator>
<dc:creator>Cavacini, L.</dc:creator>
<dc:creator>Choudhary, A. K.</dc:creator>
<dc:creator>Proulx, M. K.</dc:creator>
<dc:creator>Sundling, C.</dc:creator>
<dc:creator>Källenius, G.</dc:creator>
<dc:creator>Reljic, R.</dc:creator>
<dc:creator>Ernst, J. D.</dc:creator>
<dc:creator>Casadevall, A.</dc:creator>
<dc:creator>Locht, C.</dc:creator>
<dc:creator>Pinter, A.</dc:creator>
<dc:creator>Sassetti, C. M.</dc:creator>
<dc:creator>Bryson, B. D.</dc:creator>
<dc:creator>Fortune, S. M.</dc:creator>
<dc:creator>Alter, G.</dc:creator>
<dc:date>2024-10-11</dc:date>
<dc:identifier>doi:10.1101/2024.10.07.617070</dc:identifier>
<dc:title><![CDATA[Antibody-Fab and -Fc features promote Mycobacterium tuberculosis restriction.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.07.617095v1?rss=1">
<title>
<![CDATA[
Opioid receptors reveal a discrete cellular mechanism of endosomal G protein activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.07.617095v1?rss=1</link>
<description><![CDATA[
Many GPCRs initiate a second phase of G protein-mediated signaling from endosomes, which inherently requires an increase in G protein activity on the endosome surface. Gs-coupled GPCRs are thought to achieve this by internalizing and allosterically activating cognate G proteins again on the endosome membrane. Here we demonstrate that the -opioid receptor (MOR), a Gi-coupled GPCR, increases endosomal G protein activity in a different way. Leveraging conformational biosensors, we resolve the subcellular activation dynamics of endogenously expressed MOR and Gi/o-subclass G proteins. We show that MOR activation triggers a transient increase of active-state Gi/o on the plasma membrane that is followed by a prolonged increase on endosomes. Contrary to the Gs-coupled GPCR paradigm, however, we show that the MOR-induced increase of active-state Gi/o on endosomes requires neither internalization of MOR nor activation of MOR in the endosome membrane. We propose a distinct and additional cellular mechanism for GPCR-triggered elevation of G protein activity on endosomes that is mediated by regulated trafficking of the activated G protein rather than its activating GPCR.
]]></description>
<dc:creator>Fisher, N. M.</dc:creator>
<dc:creator>Von Zastrow, M.</dc:creator>
<dc:date>2024-10-11</dc:date>
<dc:identifier>doi:10.1101/2024.10.07.617095</dc:identifier>
<dc:title><![CDATA[Opioid receptors reveal a discrete cellular mechanism of endosomal G protein activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.07.617121v1?rss=1">
<title>
<![CDATA[
Longitudinal multi-omics reveals pathogenic TSC2 variants disrupt developmental trajectories of human cortical organoids derived from Tuberous Sclerosis Complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.07.617121v1?rss=1</link>
<description><![CDATA[
Tuberous Sclerosis Complex (TSC), an autosomal dominant condition, is caused by heterozygous mutations in either the TSC1 or TSC2 genes, manifesting in systemic growth of benign tumors. In addition to brain lesions, neurologic sequelae represent the greatest morbidity in TSC patients. Investigations utilizing TSC1/2-knockout animal or human stem cell models suggest that TSC deficiency-causing hyper-activation of mTOR signaling might precipitate anomalous neurodevelopmental processes. However, how the pathogenic variants of TSC1/2 genes affect the longitudinal trajectory of human brain development remains largely unexplored. Here, we employed 3-dimensional cortical organoids derived from induced pluripotent stem cells (iPSCs) from TSC patients harboring TSC2 variants, alongside organoids from age- and sex-matched healthy individuals as controls. Through comprehensively longitudinal molecular and cellular analyses of TSC organoids, we found that TSC2 pathogenic variants dysregulate neurogenesis, synaptogenesis, and gliogenesis, particularly for reactive astrogliosis. The altered developmental trajectory of TSC organoids significantly resembles the molecular signatures of neuropsychiatric disorders, including autism spectrum disorders, epilepsy, and intellectual disability. Intriguingly, single cell transcriptomic analyses on TSC organoids revealed that TSC2 pathogenic variants disrupt the neuron/reactive astrocyte crosstalk within the NLGN-NRXN signaling network. Furthermore, cellular and electrophysiological assessments of TSC cortical organoids, along with proteomic analyses of synaptosomes, demonstrated that the TSC2 variants precipitate perturbations in synaptic transmission, neuronal network activity, mitochondrial translational integrity, and neurofilament formation. Notably, similar perturbations were observed in surgically resected cortical specimens from TSC patients. Collectively, our study illustrates that disease-associated TSC2 variants disrupt the neurodevelopmental trajectories through perturbations of gene regulatory networks during early cortical development, leading to mitochondrial dysfunction, aberrant neurofilament formation, impaired synaptic formation and neuronal network activity.
]]></description>
<dc:creator>Niu, W.</dc:creator>
<dc:creator>Yu, S.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Chen, R.</dc:creator>
<dc:creator>Michalski, C.</dc:creator>
<dc:creator>Jahangiri, A.</dc:creator>
<dc:creator>Zohdy, Y.</dc:creator>
<dc:creator>Chern, J. J.</dc:creator>
<dc:creator>Whitworth, T. J.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Qin, Z.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Gambello, M. J.</dc:creator>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Wen, Z.</dc:creator>
<dc:date>2024-10-11</dc:date>
<dc:identifier>doi:10.1101/2024.10.07.617121</dc:identifier>
<dc:title><![CDATA[Longitudinal multi-omics reveals pathogenic TSC2 variants disrupt developmental trajectories of human cortical organoids derived from Tuberous Sclerosis Complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.10.617637v1?rss=1">
<title>
<![CDATA[
Functional screen for mediators of onco-mRNA translation specificity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.10.617637v1?rss=1</link>
<description><![CDATA[
Oncogenic protein dosage is tightly regulated to enable cancer cells to adapt and survive. Whether this is regulated at the level of translational control and the key factors in cis and trans remain unknown. The Myc oncogene is a central paradigm of an exquisitely regulated oncogene and a major driver of pancreatic ductal adenocarcinoma (PDAC). Using a functional genome-wide CRISPRi screen in PDAC cells, we identified activators of selective MYC translation through its 5 untranslated region (5UTR) and validated four RNA binding proteins (RBPs), including epitranscriptome modifiers. Among these RBPs, our top hit was RBM42, which is highly expressed in PDAC and predicts poor survival. Combining polysome sequencing and CLIP-seq analyses, we find that RBM42 binds and selectively regulates the translation of MYC and a precise, yet vital suite of pro-oncogenic transcripts, including JUN and EGFR. Mechanistically, employing IP-mass spectrometry analysis, we find that RMB42 is a novel ribosome-associated protein (RAP). Using DMS-Seq and mutagenesis analysis, we show that RBM42 directly binds and remodels the MYC 5UTR RNA structure, facilitating the formation of the translation pre-initiation complex. Importantly, RBM42 is necessary for human PDAC cell growth and fitness and PDAC tumorigenesis in xenograft mouse models in a Myc-dependent manner in vivo. In PDAC patient samples, RBM42 expression is correlated with Myc protein levels and transcriptional activity. This work transforms our understanding of the translational code in cancer and offers a new therapeutic opening to target the expression of oncogenes.
]]></description>
<dc:creator>Kovalski, J. R.</dc:creator>
<dc:creator>Sarioglu, G.</dc:creator>
<dc:creator>Subramanyam, V.</dc:creator>
<dc:creator>Hernandez, G.</dc:creator>
<dc:creator>Rademaker, G.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>Slota, M.</dc:creator>
<dc:creator>Mohan, N.</dc:creator>
<dc:creator>Yiakis, K.</dc:creator>
<dc:creator>Liu, I.</dc:creator>
<dc:creator>Wen, K. W.</dc:creator>
<dc:creator>Kim, G. E.</dc:creator>
<dc:creator>Miglani, S.</dc:creator>
<dc:creator>Burlingame, A.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Perera, R. M.</dc:creator>
<dc:creator>Ruggero, D.</dc:creator>
<dc:date>2024-10-11</dc:date>
<dc:identifier>doi:10.1101/2024.10.10.617637</dc:identifier>
<dc:title><![CDATA[Functional screen for mediators of onco-mRNA translation specificity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.11.617939v1?rss=1">
<title>
<![CDATA[
Striatal lateral inhibition regulates action selection in a mouse model of levodopa-induced dyskinesia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.11.617939v1?rss=1</link>
<description><![CDATA[
Striatal medium spiny neurons (MSNs) integrate multiple external inputs to shape motor output. In addition, MSNs form local inhibitory synaptic connections with one another. The function of striatal lateral inhibition is unknown, but one possibility is in selecting an intended action while suppressing alternatives. Action selection is disrupted in several movement disorders, including levodopa-induced dyskinesia (LID), a complication of Parkinsons disease (PD) therapy characterized by involuntary movements. Here, we identify chronic changes in the strength of striatal lateral inhibitory synapses in a mouse model of PD/LID. These synapses are also modulated by acute dopamine signaling. Chemogenetic suppression of lateral inhibition originating from dopamine D2 receptor-expressing MSNs lowers the threshold to develop involuntary movements in vivo, supporting a role in motor control. By examining the role of lateral inhibition in basal ganglia function and dysfunction, we expand the framework surrounding the role of striatal microcircuitry in action selection.
]]></description>
<dc:creator>Twedell, E. L.</dc:creator>
<dc:creator>Bair-Marshall, C. J.</dc:creator>
<dc:creator>Girasole, A. E.</dc:creator>
<dc:creator>Scaria, L. K.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2024-10-12</dc:date>
<dc:identifier>doi:10.1101/2024.10.11.617939</dc:identifier>
<dc:title><![CDATA[Striatal lateral inhibition regulates action selection in a mouse model of levodopa-induced dyskinesia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.10.617568v1?rss=1">
<title>
<![CDATA[
A Suite of Foundation Models Captures the Contextual Interplay Between Codons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.10.617568v1?rss=1</link>
<description><![CDATA[
In the canonical genetic code, many amino acids are assigned more than one codon. Work by us and others has shown that the choice of these synonymous codon is not random, and carries regulatory and functional consequences. Existing protein foundation models ignore this context-dependent role of coding sequence in shaping the protein landscape of the cell. To address this gap, we introduce cdsFM, a suite of codon-resolution large language models, including both EnCodon and DeCodon models, with up to 1B parameters. Pre-trained on 60 million protein-coding sequences from more than 5,000 species, our models effectively learn the relationship between codons and amino acids, recapitualing the overall structure of the genetic code. In addition to outperforming state-of-the-art genomic foundation models in a variety of zero-shot and few-shot learning tasks, the larger pre-trained models were superior in predicting the choice of synonymous codons. To systematically assess the impact of synonymous codon choices on protein expression and our models ability to capture these effects, we generated a large dataset measuring overall and surface expression levels of three proteins as a function of changes in their synonymous codons. We showed that our EnCodon models could be readily fine-tuned to predict the contextual consequences of synonymous codon choices. Armed with this knowledge, we applied EnCodon to existing clinical datasets of synonymous variants, and we identified a large number of synonymous codons that are likely pathogenic, several of which we experimentally confirmed in a cellbased model. Together, our findings establish the cdsFM suite as a powerful tool for decoding the complex functional grammar underlying the choice of synonymous codons.
]]></description>
<dc:creator>Naghipourfar, M.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Howard, M.</dc:creator>
<dc:creator>Macdonald, C.</dc:creator>
<dc:creator>Saberi, A.</dc:creator>
<dc:creator>Hagen, T.</dc:creator>
<dc:creator>Mofrad, M.</dc:creator>
<dc:creator>Coyote-Maestas, W.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2024-10-13</dc:date>
<dc:identifier>doi:10.1101/2024.10.10.617568</dc:identifier>
<dc:title><![CDATA[A Suite of Foundation Models Captures the Contextual Interplay Between Codons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.09.617456v1?rss=1">
<title>
<![CDATA[
The kinetics of bedaquiline diffusion in tuberculous cavities opens a window for emergence of resistance. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.09.617456v1?rss=1</link>
<description><![CDATA[
Cavitary tuberculosis (TB) is difficult to cure and a site of relapse. Bedaquiline has been a wonder drug in the treatment of multidrug resistant TB, but emergence of resistance threatens its sustained success. To investigate the role of drug distribution in resistance development, we designed a novel laser-capture microdissection scheme to spatially resolve the penetration of bedaquiline in the necrotic center (caseum) of cavities, a recalcitrant site of disease. Working with preclinical models that form large necrotic lesions, we profiled bedaquiline and two next generation diarylquinolines TBAJ-587 and TBAJ-876. Drug concentrations were measured in laser-captured areas of cavity caseum as a function of time and distance from blood supply. To simulate drug coverage in patient cavities, the data were modeled, and drug penetration parameter estimates were linked to clinical plasma pharmacokinetics for bedaquiline and the new diarylquinolines. Pharmacokinetic-pharmacodynamic (PK-PD) simulations revealed that bedaquiline reaches efficacious concentrations in outer and deep caseum after several weeks to months and lingers at subtherapeutic concentrations up to 3 years after therapy ends. TBAJ-587 and TBAJ-876, currently in clinical development, achieve bactericidal concentrations in caseum more rapidly and shorten the window of suboptimal concentrations post treatment compared to bedaquiline. Simulations of clinically plausible dosing schemes were conducted to guide the design of clinical trials for cavitary TB and help mitigate resistance development. In summary, the slow kinetics of diffusion of bedaquiline into and out of cavity caseum creates spatio-temporal windows of subtherapeutic concentrations. Site-of-disease simulations of TBAJ-587 and TBAJ-876 predict reduced opportunities for resistance development.

SIGNIFICANCEClinical resistance to bedaquiline has emerged faster than anticipated. Understanding potential contributing factors could help curb further resistance development, not only for bedaquiline but also for the next generation diarylquinolines currently in phase 2, TBAJ-587 and TBAJ-876. Here we hypothesized and confirmed that the extended time to reach steady state and slow clearance of bedaquiline leads to extremely slow diffusion into and out of cavity caseum, a recalcitrant site of TB disease and relapse. Through modeling of experimental data in a preclinical model of cavitary TB and clinical simulations, we show that the next generation diarylquinolines may reduce spatio-temporal windows of resistance development compared to bedaquiline. Our results can inform dosing schemes of diarylquinoline-based therapies that limit resistance development.
]]></description>
<dc:creator>Bustion, A. E.</dc:creator>
<dc:creator>Ernest, J. P.</dc:creator>
<dc:creator>Kaya, F.</dc:creator>
<dc:creator>Silva, C.</dc:creator>
<dc:creator>Sarathy, J.</dc:creator>
<dc:creator>Blanc, L.</dc:creator>
<dc:creator>Imperial, M.</dc:creator>
<dc:creator>Gengenbacher, M.</dc:creator>
<dc:creator>Xie, M.</dc:creator>
<dc:creator>Zimmerman, M.</dc:creator>
<dc:creator>Robertson, G. T.</dc:creator>
<dc:creator>Weiner, D.</dc:creator>
<dc:creator>Via, L. E.</dc:creator>
<dc:creator>Barry, C. E.</dc:creator>
<dc:creator>Savic, R. M.</dc:creator>
<dc:creator>Dartois, V.</dc:creator>
<dc:date>2024-10-14</dc:date>
<dc:identifier>doi:10.1101/2024.10.09.617456</dc:identifier>
<dc:title><![CDATA[The kinetics of bedaquiline diffusion in tuberculous cavities opens a window for emergence of resistance.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.11.617955v1?rss=1">
<title>
<![CDATA[
Human brain changes after first psilocybin use 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.11.617955v1?rss=1</link>
<description><![CDATA[
Psychedelics have robust effects on acute brain function and long-term behavior but whether they also cause enduring functional and anatomical brain changes is unknown. In a placebo-controlled, within-subjects, electroencephalography, and magnetic resonance imaging study in 28 healthy, entirely psychedelic-naive participants, anatomical and functional brain changes were detected from one-hour to one-month after a single high-dose (25 mg) of psilocybin. Increases in cognitive flexibility, psychological insight, and well-being were seen at one-month. Diffusion imaging done before and one-month after 25mg psilocybin revealed decreased axial diffusivity bilaterally in prefrontal-subcortical tracts that correlated with decreased brain network modularity over the same time period. Decreased modularity also correlated with improved well-being. Increased cortical signal entropy at 1- and 2-hours post-dosing predicted improved psychological well-being at one-month. Next-day psychological insight mediated the entropy to well-being relationship. All effects were exclusive to 25mg psilocybin; no effects occurred with a 1mg psilocybin  placebo dose.
]]></description>
<dc:creator>Lyons, T.</dc:creator>
<dc:creator>Spriggs, M.</dc:creator>
<dc:creator>Kerkela, L.</dc:creator>
<dc:creator>Rosas, F.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Timmermann, C.</dc:creator>
<dc:creator>Oestreich, L.</dc:creator>
<dc:creator>Pagni, B.</dc:creator>
<dc:creator>Zeifman, R.</dc:creator>
<dc:creator>Hampshire, A.</dc:creator>
<dc:creator>Trender, W.</dc:creator>
<dc:creator>Douglas, H.</dc:creator>
<dc:creator>Girn, M.</dc:creator>
<dc:creator>Godfrey, K.</dc:creator>
<dc:creator>Kettner, H.</dc:creator>
<dc:creator>Sharif, F.</dc:creator>
<dc:creator>Espasiano, L.</dc:creator>
<dc:creator>Gazzaley, A.</dc:creator>
<dc:creator>Wall, M.</dc:creator>
<dc:creator>Erritzoe, D.</dc:creator>
<dc:creator>Nutt, D.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:date>2024-10-14</dc:date>
<dc:identifier>doi:10.1101/2024.10.11.617955</dc:identifier>
<dc:title><![CDATA[Human brain changes after first psilocybin use]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.12.618016v1?rss=1">
<title>
<![CDATA[
Immunological Profiling in Knee Osteoarthritis: Treg Dysfunction as Key Driver of Pain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.12.618016v1?rss=1</link>
<description><![CDATA[
Pain is the hallmark symptom of osteoarthritis (OA) and its biological drivers remain poorly understood. While the role of innate immunity in OA has been extensively studied, the involvement of adaptive immunity, in particular regulatory T cells (Tregs), is not well understood. Using a comprehensive multi-omic approach on the peripheral blood from 46 knee OA patients with similar radiographic stage, including deep immunophenotyping, cytokine profiling, transcriptomic and T-cell receptor analysis on sorted CD4 Tregs and effector T cells (Teff), we identified an immunological signature associated with OA-related pain. Cytokines promoting Treg expansion and activation (with increases of sIL2-RA, sTNFR1, sTNFR2) were correlated with the Western Ontario and McMaster Universities Arthritis Index (WOMAC) pain subscore, suggesting a potential Treg dysfunction. Nineteen T cell subsets were correlated with WOMAC pain. Notably, we found a negative correlation of cell subsets associated with Treg expansion and activation (FoxP3+CTLA4+, CD4+CD57+, Treg CD95+, CD4 Treg CD45RA-). Differential gene expression analysis between patients with low and high WOMAC pain intensity (threshold [&ge;] 40/100) revealed an upregulation of inflammasome-related genes such as IL1RL1, IL31RA, IFITM3, NLRP3, IFNG in Tregs. Functional enrichment analysis highlighted an overrepresentation of innate immune response, IL-8, and interferon activation pathways suggesting a pro-inflammatory state in Tregs of patients with high pain intensity. Collectively, our systems immunology approach highlights multiple associations between Treg dysfunctionality and OA-related pain, providing new insights into the adaptive immune systems contribution to OA-related pain.
]]></description>
<dc:creator>Binvignat, M.</dc:creator>
<dc:creator>Dubois, J.</dc:creator>
<dc:creator>Marco Salvador, M.</dc:creator>
<dc:creator>Stys, P.</dc:creator>
<dc:creator>Pitoiset, F.</dc:creator>
<dc:creator>Roux, A.</dc:creator>
<dc:creator>Barbie, M.</dc:creator>
<dc:creator>Hassler, S.</dc:creator>
<dc:creator>Lorenzon, R.</dc:creator>
<dc:creator>Ribet, C.</dc:creator>
<dc:creator>Mhanna, V.</dc:creator>
<dc:creator>Vantomme, H.</dc:creator>
<dc:creator>Adda, L.</dc:creator>
<dc:creator>Barennes, P.</dc:creator>
<dc:creator>Coatnoan, N.</dc:creator>
<dc:creator>Legouge, K.</dc:creator>
<dc:creator>Aheng, C.</dc:creator>
<dc:creator>Courties, A.</dc:creator>
<dc:creator>Minssen, L.</dc:creator>
<dc:creator>Butte, A. J.</dc:creator>
<dc:creator>Six, A.</dc:creator>
<dc:creator>Rosenzwajg, M.</dc:creator>
<dc:creator>Tchitchek, N.</dc:creator>
<dc:creator>Berenbaum, F.</dc:creator>
<dc:creator>Klatzmann, D.</dc:creator>
<dc:creator>Mariotti-Ferrandiz, E.</dc:creator>
<dc:creator>Sellam, J.</dc:creator>
<dc:date>2024-10-15</dc:date>
<dc:identifier>doi:10.1101/2024.10.12.618016</dc:identifier>
<dc:title><![CDATA[Immunological Profiling in Knee Osteoarthritis: Treg Dysfunction as Key Driver of Pain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.14.616932v1?rss=1">
<title>
<![CDATA[
The Taiwan Precision Medicine Initiative: A Cohort for Large-Scale Studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.14.616932v1?rss=1</link>
<description><![CDATA[
The Taiwan Precision Medicine Initiative (TPMI), a project initiated by the Academia Sinica in collaboration with 16 major medical centers around Taiwan, has recruited 565,390 participants who consented to provide DNA samples for genetic profiling and grant access to their electronic medical records (EMR) for studies to develop precision medicine. Access to the EMR is both retrospective and prospective, allowing researchers to conduct prospective studies over time. Genetic profiling is done with population-optimized SNP arrays for the Han Chinese populations that enable genetic analyses such as genome-wide association, phenome-wide association, and polygenic risk score studies to evaluate common disease risk and pharmacogenetic response. Furthermore, the TPMI participants agree to be contacted for future research opportunities related to their genetic risks and receive personalized genetic risk profiles with health management recommendations. TPMI has established the TPMI Data Access Platform (TDAP), a central database and analysis platform that both safeguards the security of the data and facilitates academic research. The TPMI is the largest non-European cohort that merges genetic profiles with EMR in the world. With a cohort that can be followed over time, it can be utilized to validate genetic risk prediction models, conduct clinical trials to show the efficacy of risk-based health management, and optimize health policies based on genetic risks. In this report, we describe the TPMI study design, the population and genetic characteristics of the TPMI cohort, and the power it provides to conduct crucial studies in developing precision medicine on a population and personal level. As Han Chinese represent almost 20% of the worlds population, the results of TPMI studies will benefit >1.4 billion people around the world and serve as a model for developing population-based precision medicine.
]]></description>
<dc:creator>Yang, H.-C.</dc:creator>
<dc:creator>Kwok, P.-Y.</dc:creator>
<dc:creator>Li, L.-H.</dc:creator>
<dc:creator>Liu, Y.-M.</dc:creator>
<dc:creator>Jong, Y.-J.</dc:creator>
<dc:creator>Lee, K.-Y.</dc:creator>
<dc:creator>Wang, D.-W.</dc:creator>
<dc:creator>Tsai, M.-F.</dc:creator>
<dc:creator>Yang, J.-H.</dc:creator>
<dc:creator>Chen, C.-H.</dc:creator>
<dc:creator>Yeh, E.-C.</dc:creator>
<dc:creator>Wei, C.-y.</dc:creator>
<dc:creator>Fann, C. S.- J.</dc:creator>
<dc:creator>Huang, Y.-T.</dc:creator>
<dc:creator>Chen, C.-W.</dc:creator>
<dc:creator>Lee, Y.-J.</dc:creator>
<dc:creator>Chu, S.-K.</dc:creator>
<dc:creator>Ho, C.-h.</dc:creator>
<dc:creator>Yang, C.-S.</dc:creator>
<dc:creator>Lee, Y. L.</dc:creator>
<dc:creator>Chen, H.-H.</dc:creator>
<dc:creator>Hou, M.-C.</dc:creator>
<dc:creator>Chiou, J.-F.</dc:creator>
<dc:creator>Yang, S.-F.</dc:creator>
<dc:creator>Wang, C.-H.</dc:creator>
<dc:creator>Huang, C.-Y.</dc:creator>
<dc:creator>Chiu, K.-M.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Chiang, F.-T.</dc:creator>
<dc:creator>Lee, S.-L.</dc:creator>
<dc:creator>Chen, S.-S.</dc:creator>
<dc:creator>Yao, W.-J.</dc:creator>
<dc:creator>Chien, C.-C.</dc:creator>
<dc:creator>Lin, S.-Y.</dc:creator>
<dc:creator>Chang, F.-P.</dc:creator>
<dc:creator>Ho, H.-L.</dc:creator>
<dc:creator>Yeh, Y.-C.</dc:creator>
<dc:creator>Tseng, W.-C.</dc:creator>
<dc:creator>Lin, M.-H.</dc:creator>
<dc:creator>Chang, H.-T.</dc:creator>
<dc:creator>Tseng, L.-M.</dc:creator>
<dc:creator>Liang, W.-Y.</dc:creator>
<dc:creator>Chen, P. C.-H.</dc:creator>
<dc:creator>Hang, J.-F.</dc:creator>
<dc:creator>Lin, S.-C.</dc:creator>
<dc:creator>Chan, Y.-J.</dc:creator>
<dc:creator>Kuo, Y.</dc:creator>
<dc:date>2024-10-17</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.616932</dc:identifier>
<dc:title><![CDATA[The Taiwan Precision Medicine Initiative: A Cohort for Large-Scale Studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.14.618095v1?rss=1">
<title>
<![CDATA[
Replicating retroviral delivery of an IL-15 superagonist improves antitumor immunity and long-term survival in poorly immunogenic glioblastoma models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.14.618095v1?rss=1</link>
<description><![CDATA[
Glioblastoma (GBM) is the most lethal primary brain neoplasm due to its highly immunosuppressive microenvironment and resistance to conventional therapies. To overcome this challenge, we engineered a replicating retrovirus (RRV) to deliver a superagonist interleukin-15 receptor-linked fusion protein (RLI) directly to tumor cells, engineering them into local immunotherapy biofactories. This strategy leverages the tumor-selective replication of RRV to achieve localized and sustained RLI expression within the tumor microenvironment. In two orthotopic poorly immunogenic GBM mouse models, intratumoral administration of RRV RLI significantly reduced tumor growth and prolonged survival compared to controls, with some mice achieving long-term remission and demonstrating immunologic memory upon rechallenge. Transcriptomic and flow cytometric analyses revealed that RRV RLI treatment enhanced infiltration and activation of CD8 T cells, NK cells, and upregulated antigen presentation pathways within the tumor microenvironment. Depletion studies indicated that the therapeutic efficacy of RRV RLI is dependent on both CD4 and CD8 T cells. Notably, combining RRV RLI with the GBM standard of care chemotherapeutic agent temozolomide (TMZ) synergistically improved survival outcomes. Subsequent single-cell RNA and T cell receptor sequencing identified enhanced effector cell activation, antigen presentation, and clonal T cell expansion in the combination therapy group. Further T cell receptor analysis and clustering implied a tumor-specific immune response rather than one targeting the viral delivery vehicle, suggesting that this therapeutic approach could be reapplied without eliciting anti-vector immunity. Our findings suggest that RRV-mediated delivery of RLI effectively transforms GBM tumors into immunostimulatory hubs, eliciting a potent anti-tumor immune response. This novel viral immunotherapy holds significant promise for clinical translation in the treatment of GBM and other difficult-to-treat solid tumors.
]]></description>
<dc:creator>Haddad, A. F.</dc:creator>
<dc:creator>Saha, A.</dc:creator>
<dc:creator>Spatz, J.</dc:creator>
<dc:creator>Collins, S.</dc:creator>
<dc:creator>Lovalvo, I.</dc:creator>
<dc:creator>Gill, S.</dc:creator>
<dc:creator>Montoya, M. L.</dc:creator>
<dc:creator>Shukla, P.</dc:creator>
<dc:creator>Hong, J.</dc:creator>
<dc:creator>Wang, E.</dc:creator>
<dc:creator>Chuntova, P.</dc:creator>
<dc:creator>Lad, M.</dc:creator>
<dc:creator>Chalif, E.</dc:creator>
<dc:creator>Kasahara, N.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:date>2024-10-17</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618095</dc:identifier>
<dc:title><![CDATA[Replicating retroviral delivery of an IL-15 superagonist improves antitumor immunity and long-term survival in poorly immunogenic glioblastoma models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.14.618111v1?rss=1">
<title>
<![CDATA[
Telocytes link epithelial nutrient sensing with amplification of the ILC2-tuft cell circuit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.14.618111v1?rss=1</link>
<description><![CDATA[
Group 2 innate lymphocytes (ILC2s) are prevalent in small intestine but engagement of type 2 immunity during basal processes are incompletely described. Thymic stromal lymphopoietin (TSLP), a cytokine implicated in ILC2 activation, was constitutively expressed in villus telocytes and crypt-associated trophocytes, specialized fibroblasts that sustain epithelial identity. Feeding increased TSLP and induced ILC2 type 2 cytokines that were attenuated by deletion of TSLP in PDGFR+ stromal cells or TSLP receptor on ILC2s. Mouse and human telocytes expressed receptors for glucagon-like peptide-2 (GLP-2), which is released by enteroendocrine cells (EECs) after eating. GLP-2 induced intestinal TSLP, TSLP-dependent ILC2 cytokine production, and tuft cell hyperplasia. The telocyte-alarmin relay couples EEC nutrient detection with amplification of a tuft cell chemosensory circuit that diversifies surveillance of ingested cargo.

One-Sentence SummaryIntestinal telocyte TSLP relays signals from enteroendocrine cells to ILC2s to amplify the tuft cell circuit in response to feeding.
]]></description>
<dc:creator>Liao, C.</dc:creator>
<dc:creator>Ji, M.</dc:creator>
<dc:creator>Wang, Z.-E.</dc:creator>
<dc:creator>Drucker, D. J.</dc:creator>
<dc:creator>Liang, H.-E.</dc:creator>
<dc:creator>Locksley, R. M.</dc:creator>
<dc:date>2024-10-17</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618111</dc:identifier>
<dc:title><![CDATA[Telocytes link epithelial nutrient sensing with amplification of the ILC2-tuft cell circuit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.14.618280v1?rss=1">
<title>
<![CDATA[
Representations of stimulus meaning in the hippocampus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.14.618280v1?rss=1</link>
<description><![CDATA[
The ability to discriminate and categorize the meaning of environmental stimuli and respond accordingly is essential for survival. The ventral hippocampus (vHPC) controls emotional and motivated behaviors in response to environmental cues and is hypothesized to do so in part by deciphering the positive or negative quality of these cues. Yet, what features of the environment are represented in the activity patterns of vCA1 neurons, and whether the positive or negative meaning of a stimulus is present at this stage, remains unclear. Here, using 2-photon calcium imaging across six different experimental paradigms, we consistently found that vCA1 ensembles encode the identity, sensory features, and intensity of learned and innately salient stimuli, but not their overall valence. These results offer a reappraisal of vCA1 function, wherein information corresponding to individual stimulus features and their behavioral saliency predominates, while valence-related information is attached elsewhere.
]]></description>
<dc:creator>Biane, J. S.</dc:creator>
<dc:creator>Ladow, M. A.</dc:creator>
<dc:creator>Fan, A.</dc:creator>
<dc:creator>Choi, H. S.</dc:creator>
<dc:creator>Zhou, L. Z.</dc:creator>
<dc:creator>Hassan, S.</dc:creator>
<dc:creator>Apodaca-Montano, D. L.</dc:creator>
<dc:creator>Kwon, A. O.</dc:creator>
<dc:creator>Bratsch-Prince, J. X.</dc:creator>
<dc:creator>Kheirbek, M. A.</dc:creator>
<dc:date>2024-10-17</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618280</dc:identifier>
<dc:title><![CDATA[Representations of stimulus meaning in the hippocampus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.15.618087v1?rss=1">
<title>
<![CDATA[
Dark Microglia Are Abundant in Normal Postnatal Development, where they Remodel Synapses via Phagocytosis and Trogocytosis, and Are Dependent on TREM2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.15.618087v1?rss=1</link>
<description><![CDATA[
This study examined dark microglia--a state linked to central nervous system pathology and neurodegeneration--during postnatal development in the mouse ventral hippocampus, finding that dark microglia interact with blood vessels and synapses and perform trogocytosis of pre-synaptic axon terminals. Furthermore, we found that dark microglia in development notably expressed C-type lectin domain family 7 member A (CLEC7a), lipoprotein lipase (LPL) and triggering receptor expressed on myeloid cells 2 (TREM2) and required TREM2, differently from other microglia, suggesting a link between their role in remodeling during development and central nervous system pathology. Together, these results point towards a previously under-appreciated role for dark microglia in synaptic pruning and plasticity during normal postnatal development.
]]></description>
<dc:creator>Vecchiarelli, H. A.</dc:creator>
<dc:creator>Bisht, K.</dc:creator>
<dc:creator>Sharma, K. P.</dc:creator>
<dc:creator>Weiser Novak, S.</dc:creator>
<dc:creator>Traetta, M. E.</dc:creator>
<dc:creator>Garcia-Segura, M. E.</dc:creator>
<dc:creator>St-Pierre, M.-K.</dc:creator>
<dc:creator>Savage, J. C.</dc:creator>
<dc:creator>Willis, C.</dc:creator>
<dc:creator>Picard, K.</dc:creator>
<dc:creator>Bordeleau, M.</dc:creator>
<dc:creator>Vernoux, N.</dc:creator>
<dc:creator>Khakpour, M.</dc:creator>
<dc:creator>Garg, R.</dc:creator>
<dc:creator>Loewen, S. M.</dc:creator>
<dc:creator>Murray, C. J.</dc:creator>
<dc:creator>Grinberg, Y. Y.</dc:creator>
<dc:creator>Faustino, J.</dc:creator>
<dc:creator>Halvorson, T.</dc:creator>
<dc:creator>Lau, V.</dc:creator>
<dc:creator>Pluchino, S.</dc:creator>
<dc:creator>Vexler, Z. S.</dc:creator>
<dc:creator>Carson, M. J.</dc:creator>
<dc:creator>Manor, U.</dc:creator>
<dc:creator>Peruzzotti-Jametti, L.</dc:creator>
<dc:creator>Tremblay, M.-E.</dc:creator>
<dc:date>2024-10-18</dc:date>
<dc:identifier>doi:10.1101/2024.10.15.618087</dc:identifier>
<dc:title><![CDATA[Dark Microglia Are Abundant in Normal Postnatal Development, where they Remodel Synapses via Phagocytosis and Trogocytosis, and Are Dependent on TREM2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.15.618329v1?rss=1">
<title>
<![CDATA[
Adhesion G protein-coupled receptor ADGRG1 promotes protective microglial response in Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.15.618329v1?rss=1</link>
<description><![CDATA[
Germline genetic architecture of Alzheimers disease (AD) indicates microglial mechanisms of disease susceptibility and outcomes. However, the mechanisms that enable microglia to mediate protective responses to AD pathology remain elusive. Adgrg1 is specifically expressed in yolk-sac-derived microglia. This study reveals the role of yolk-sac-derived microglia in AD pathology, highlighting the function of ADGRG1 in modulating microglial protective responses to amyloid deposition. Utilizing both constitutive and inducible microglial Adgrg1 knockout 5xFAD models, we demonstrate that Adgrg1 deficiency leads to increased amyloid deposition, exacerbated neuropathology, and accelerated cognitive impairment. Transcriptomic analyses reveal a distinct microglial state characterized by downregulated genes associated with homeostasis, phagocytosis, and lysosomal functions. Functional assays in mouse models and human embryonic stem cells-derived microglia support that microglial ADGRG1 is required for efficient A{beta} phagocytosis. Together, these results uncover a GPCR-dependent microglial response to A{beta}, pointing towards potential therapeutic strategies to alleviate disease progression by enhancing microglial functional competence.
]]></description>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Wangzhou, A.</dc:creator>
<dc:creator>Yu, D.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Schmidt, R.</dc:creator>
<dc:creator>De Florencio, S. L.</dc:creator>
<dc:creator>Chao, L.</dc:creator>
<dc:creator>Perez, Y.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Ransohoff, R. M.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Piao, X.</dc:creator>
<dc:date>2024-10-18</dc:date>
<dc:identifier>doi:10.1101/2024.10.15.618329</dc:identifier>
<dc:title><![CDATA[Adhesion G protein-coupled receptor ADGRG1 promotes protective microglial response in Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.15.618585v1?rss=1">
<title>
<![CDATA[
CELL-Diff: Unified diffusion modeling for protein sequences and microscopy images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.15.618585v1?rss=1</link>
<description><![CDATA[
Fluorescence microscopy is ubiquitously used in cell biology research to characterize the cellular role of a protein. To help elucidate the relationship between the amino acid sequence of a protein and its cellular function, we introduce CELL-Diff, a unified diffusion model facilitating bidirectional transformations between protein sequences and their corresponding microscopy images. Utilizing reference cell morphology images and a protein sequence, CELL-Diff efficiently generates corresponding protein images. Conversely, given a protein image, the model out-puts protein sequences. CELL-Diff integrates continuous and diffusion models within a unified framework and is implemented using a transformer-based network. We train CELL-Diff on the Human Protein Atlas (HPA) dataset and finetune it on the OpenCell dataset. Experimental results demonstrate that CELL-Diff outperforms existing methods in generating high-fidelity protein images, making it a practical tool for investigating subcellular protein localization and interactions.
]]></description>
<dc:creator>Zheng, D.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:date>2024-10-18</dc:date>
<dc:identifier>doi:10.1101/2024.10.15.618585</dc:identifier>
<dc:title><![CDATA[CELL-Diff: Unified diffusion modeling for protein sequences and microscopy images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.16.618588v1?rss=1">
<title>
<![CDATA[
Methylation Clocks Do Not Predict Age or Alzheimer's Disease Risk Across Genetically Admixed Individuals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.16.618588v1?rss=1</link>
<description><![CDATA[
Epigenetic aging clocks based on DNA methylation patterns across the genome have emerged as a potential biomarker for risk of age-related diseases, like Alzheimers disease (AD), and environmental and social stressors. However, methylation clocks have not been comprehensively validated in genetically diverse individuals. Here we evaluate a set of first-, second-, and third-generation methylation clocks in 621 AD patients and matched controls from African American, Hispanic, and White cohorts. The clocks are less accurate at predicting age in genetically admixed cohorts compared to the White cohort, especially for those with substantial African ancestry. This decreased accuracy holds in >2,500 individuals of European and African ancestry from three additional datasets. The clocks also fail to consistently identify age acceleration in admixed AD cases compared to controls. To explore potential causes for the lack of generalization of the clocks, we intersected clock CpGs with methylation, germline genetic variants, and methylation QTL (meQTL) data from global populations. We find differential methylation between African and European ancestry individuals is common for clock CpGs. Genetic variants rarely disrupt clock CpGs between populations, but a substantial fraction of clock CpGs have meQTL with significantly higher frequencies in African genetic ancestries. Our results demonstrate that methylation clocks often fail to predict age and AD risk when applied across populations and suggest avenues for improving their portability by considering differences in genetic and epigenetic patterns across human populations.
]]></description>
<dc:creator>Cruz-Gonzalez, S.</dc:creator>
<dc:creator>Gu, E.</dc:creator>
<dc:creator>Gomez, L.</dc:creator>
<dc:creator>Mews, M.</dc:creator>
<dc:creator>Vance, J. M.</dc:creator>
<dc:creator>Cuccaro, M. L.</dc:creator>
<dc:creator>Cornejo-Olivas, M. R.</dc:creator>
<dc:creator>Feliciano-Astacio, B. E.</dc:creator>
<dc:creator>Byrd, G. S.</dc:creator>
<dc:creator>Haines, J. L.</dc:creator>
<dc:creator>Pericak-Vance, M. A.</dc:creator>
<dc:creator>Griswold, A. J.</dc:creator>
<dc:creator>Bush, W. S.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:date>2024-10-18</dc:date>
<dc:identifier>doi:10.1101/2024.10.16.618588</dc:identifier>
<dc:title><![CDATA[Methylation Clocks Do Not Predict Age or Alzheimer's Disease Risk Across Genetically Admixed Individuals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.19.619180v1?rss=1">
<title>
<![CDATA[
Microglia regulate GABAergic neurogenesis in prenatal human brain through IGF1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.19.619180v1?rss=1</link>
<description><![CDATA[
GABAergic neurons are an essential cellular component of neural circuits. Their abundance and diversity have enlarged significantly in the human brain, contributing to the expanded cognitive capacity of humans. However, the developmental mechanism of the extended production of GABAergic neurons in the human brain remains elusive. Here, we use single-cell transcriptomics, bioinformatics, and histological analyses to uncover microglial regulation of the sustained proliferation of GABAergic progenitors and neuroblasts in the human medial ganglionic eminence (hMGE). We show that insulin-like growth factor 1 (IGF1) and its receptor IGR1R as the top ligand-receptor pair underlying microglia-progenitor communication in the prenatal human brain. Using our newly developed neuroimmune hMGE organoids, which mimics hMGE cytoarchitecture and developmental trajectory, we demonstrate that microglia-derived IGF1 promotes progenitor proliferation and the production of GABAergic neurons. Conversely, IGF1-neutralizing antibodies and IGF1 knockout human embryonic stem cells (hESC)-induced microglia (iMG) completely abolished iMG-mediated progenitor proliferation. Together, these findings reveal a previously unappreciated role of microglia-derived IGF1 in promoting proliferation of neural progenitors and the development of GABAergic neurons.
]]></description>
<dc:creator>Yu, D.</dc:creator>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Wangzhou, A.</dc:creator>
<dc:creator>De Florencio, S.</dc:creator>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Paredes, M.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Piao, X.</dc:creator>
<dc:date>2024-10-20</dc:date>
<dc:identifier>doi:10.1101/2024.10.19.619180</dc:identifier>
<dc:title><![CDATA[Microglia regulate GABAergic neurogenesis in prenatal human brain through IGF1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.20.619312v1?rss=1">
<title>
<![CDATA[
Medicinal chemistry of benzoisoxazole-4,7-dione analogues identifies the ribosomal small subunit as a target for specific mycobacterial translational fidelity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.20.619312v1?rss=1</link>
<description><![CDATA[
A central dogma of molecular biology is the "speed-accuracy trade-off," where ribosomes must slow down to ensure accurate protein synthesis. In mycobacteria, a high basal level of mistranslation at glutamine and asparagine codons, caused by an indirect tRNA aminoacylation pathway, promotes tolerance to the antibiotic rifampicin. While pharmacologically increasing translational fidelity is a promising strategy to combat antibiotic tolerance, the underlying mechanisms remain poorly understood. Here, we screened 9,000 synthetic compounds and identified benzo[d]isoxazole-4,7-diones as a novel chemical class that reduces mycobacterial mistranslation. Medicinal chemistry optimization yielded a lead compound, 9787, with superior potency in decreasing mistranslation and reversing rifampicin tolerance. Using competitive chemical proteomics, we identified the 30S ribosomal protein S5 (RpS5) as the specific cellular target. Remarkably, compound 9787 enhances translational fidelity at concentrations that do not measurably impact the overall rate of protein synthesis. Our findings challenge the universality of the speed-accuracy trade-off, demonstrating that fidelity can be improved independently of translation speed. This work reveals that the ribosomal small subunit is a druggable target for modulating translational quality control and introduces a new strategy for combating antibiotic-tolerant bacteria without the associated fitness cost of slowed translation.

ImportanceA fundamental principle in molecular biology holds that ribosomes face a trade-off between translation speed and accuracy: going faster means making more errors, while maintaining high fidelity requires slowing down. This study challenges that paradigm by identifying a small molecule that increases translational accuracy in mycobacteria without affecting the rate of protein synthesis. The compound targets ribosomal protein S5 and specifically reduce errors arising from physiologically mischarged tRNAs - a quality control problem distinct from the well-studied codon*anticodon mismatches. This form of mistranslation contributes to antibiotic tolerance in tuberculosis, making it a potential therapeutic target. Our findings reveal unexpected flexibility in how ribosomes maintain translation quality and suggest that pharmacologically increasing fidelity without the fitness cost of slowed protein synthesis may be an attractive strategy for combating antibiotic-tolerant bacteria.
]]></description>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Chaudhuri, S.</dc:creator>
<dc:creator>Feid, S. C.</dc:creator>
<dc:creator>Pan, M.</dc:creator>
<dc:creator>Kawaji, Q.</dc:creator>
<dc:creator>Liu, G.</dc:creator>
<dc:creator>Javid, B.</dc:creator>
<dc:date>2024-10-21</dc:date>
<dc:identifier>doi:10.1101/2024.10.20.619312</dc:identifier>
<dc:title><![CDATA[Medicinal chemistry of benzoisoxazole-4,7-dione analogues identifies the ribosomal small subunit as a target for specific mycobacterial translational fidelity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.20.619055v1?rss=1">
<title>
<![CDATA[
Shc1 cooperates with Frs2 and Shp2 to recruit Grb2 in FGF-induced lens development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.20.619055v1?rss=1</link>
<description><![CDATA[
Fibroblast growth factor (FGF) signaling elicits multiple downstream pathways, most notably the Ras/MAPK cascade facilitated by the adaptor protein Grb2. However, the mechanism by which Grb2 is recruited to the FGF signaling complex remains unresolved. Here we showed that genetic ablation of FGF signaling prevented lens induction by disrupting transcriptional regulation and actin cytoskeletal arrangements, which could be reproduced by deleting the juxtamembrane region of the FGF receptor and rescued by Kras activation. Conversely, mutations affecting the Frs2-binding site on the FGF receptor or the deletion of Frs2 and Shp2 primarily impact later stages of lens vesicle development involving lens fiber cell differentiation. Our study further revealed that the loss of Grb2 abolished MAPK signaling, resulting in a profound arrest of lens development. However, removing Grb2s putative Shp2 dephosphorylation site (Y209) neither produced a detectable phenotype nor impaired MAPK signaling during lens development. Furthermore, the catalytically inactive Shp2 mutation (C459S) only modestly impaired FGF signaling, whereas replacing Shp2s C-terminal phosphorylation sites (Y542/Y580) previously implicated in Grb2 binding only caused placental defects, perinatal lethality, and reduced lacrimal gland branching without impacting lens development, suggesting that Shp2 only partially mediates Grb2 recruitment. In contrast, we observed that FGF signaling is required for the phosphorylation of the Grb2-binding sites on Shc1 and the deletion of Shc1 exacerbates the lens vesicle defect caused by Frs2 and Shp2 deletion. These findings establish Shc1 as a critical collaborator with Frs2 and Shp2 in targeting Grb2 during FGF signaling.
]]></description>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Garg, A.</dc:creator>
<dc:creator>Park, E. S.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Chow, A.</dc:creator>
<dc:creator>Peregrin, J.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.20.619055</dc:identifier>
<dc:title><![CDATA[Shc1 cooperates with Frs2 and Shp2 to recruit Grb2 in FGF-induced lens development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.21.618927v1?rss=1">
<title>
<![CDATA[
Differential enrichment of retinal ganglion cells underlies proposed core neurodegenerative transcription programs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.21.618927v1?rss=1</link>
<description><![CDATA[
In a published Correction1, a revised analysis updated two "core transcription programs" proposed to underlie axon injury-induced retinal ganglion cell (RGC) neurodegeneration. Though extensive, the Correction purported to leave the two principal conclusions of its parent study2 unaltered. The first of those findings was that a core program mediated by the Activating Transcription Factor-4 (ATF4) and its likely heterodimeric partner does not include numerous canonical ATF4 target genes stimulated by RGC axon injury. The second was that the Activating Transcription Factor-3 (ATF3) and C/EBP Homologous Protein (CHOP) function with unprecedented coordination in a parallel program regulating innate immunity pathways. Here those unexpected findings are revealed to instead reflect insufficient knockout coupled with differences in RGC enrichment across conditions. This analysis expands on the published Corrections redefinition of the purported transcription programs to raise foundational questions about the proposed functions and relationships of these transcription factors in neurodegeneration.
]]></description>
<dc:creator>Watkins, T. A.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.21.618927</dc:identifier>
<dc:title><![CDATA[Differential enrichment of retinal ganglion cells underlies proposed core neurodegenerative transcription programs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.22.619677v1?rss=1">
<title>
<![CDATA[
Phenotypic landscape of a fungal meningitis pathogen reveals its unique biology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.22.619677v1?rss=1</link>
<description><![CDATA[
Cryptococcus neoformans is the most common cause of fungal meningitis and the top-ranked W.H.O. priority fungal pathogen. Only distantly related to model fungi, C. neoformans is also a powerful experimental system for exploring conserved eukaryotic mechanisms lost from specialist model yeast lineages. To decipher its biology globally, we constructed 4328 gene deletions and measured--with exceptional precision--the fitness of each mutant under 141 diverse growth-limiting in vitro conditions and during murine infection. We defined functional modules by clustering genes based on their phenotypic signatures. In-depth studies leveraged these data in two ways. First, we defined and investigated new components of key signaling pathways, which revealed animal-like pathways/components not predicted from studies of model yeasts. Second, we identified environmental adaptation mechanisms repurposed to promote mammalian virulence by C. neoformans, which lacks a known animal reservoir. Our work provides an unprecedented resource for deciphering a deadly human pathogen.
]]></description>
<dc:creator>Boucher, M.</dc:creator>
<dc:creator>Banerjee, S.</dc:creator>
<dc:creator>Joshi, M.</dc:creator>
<dc:creator>Wei, A.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Lei, S.</dc:creator>
<dc:creator>Ciranni, M.</dc:creator>
<dc:creator>Condon, A.</dc:creator>
<dc:creator>Langen, A.</dc:creator>
<dc:creator>Goddard, T.</dc:creator>
<dc:creator>Caradonna, I.</dc:creator>
<dc:creator>Goranov, A.</dc:creator>
<dc:creator>Homer, C.</dc:creator>
<dc:creator>Mortensen, Y.</dc:creator>
<dc:creator>Petnic, S.</dc:creator>
<dc:creator>Reilly, M.</dc:creator>
<dc:creator>Xiong, Y.</dc:creator>
<dc:creator>Susa, K.</dc:creator>
<dc:creator>Pastore, V. P.</dc:creator>
<dc:creator>Zaro, B.</dc:creator>
<dc:creator>Madhani, H.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.619677</dc:identifier>
<dc:title><![CDATA[Phenotypic landscape of a fungal meningitis pathogen reveals its unique biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.21.619520v1?rss=1">
<title>
<![CDATA[
Multigenerational Proteolytic Inactivation of Restriction Upon Subtle Genomic Hypomethylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.21.619520v1?rss=1</link>
<description><![CDATA[
Restriction-modification (R-M) systems, present in most bacterial genomes, protect against phage infection by detecting and degrading invading foreign DNA. However, like many prokaryotic anti-phage systems, R-M systems pose a significant risk of auto-immunity, exacerbated by the presence of hundreds to thousands of potential cleavage sites in the bacterial genome. In Pseudomonas aeruginosa, restriction inactivation upon growth at high temperatures was previously described, however, which system is being inactivated, the underlying mechanism, as well as the timing of recovery, remain unknown. Here, we report that P. aeruginosa Type I methyltransferase (HsdMS) and restriction endonuclease (HsdR) components are degraded by two Lon-like proteases when replicating above 41 {degrees}C, which induces partial genome hypomethylation and simultaneously prevents self-targeting, respectively. Interestingly, upon return to 37 {degrees}C, methyltransferase activity returns gradually, with restriction activity not fully recovering for over 60 bacterial generations, representing the longest bacterial memory to our knowledge. Forced expression of HsdR over the first 45 generations is toxic, demonstrating the fitness benefit of HsdR inactivation. Our findings demonstrate that type I R-M is tightly regulated post-translationally with a remarkable memory effect to ensure genomic stability and emphasize the importance of mitigating auto-toxicity for bacterial defense systems.
]]></description>
<dc:creator>Shmidov, E.</dc:creator>
<dc:creator>Villani, A.</dc:creator>
<dc:creator>Mendoza, S. D.</dc:creator>
<dc:creator>Avihu, E.</dc:creator>
<dc:creator>Lebenthal-Loinger, I.</dc:creator>
<dc:creator>Karako-Lampert, S.</dc:creator>
<dc:creator>Shoshani, S.</dc:creator>
<dc:creator>Ye, C.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Yan, H.</dc:creator>
<dc:creator>Tang, W.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Banin, E.</dc:creator>
<dc:date>2024-10-23</dc:date>
<dc:identifier>doi:10.1101/2024.10.21.619520</dc:identifier>
<dc:title><![CDATA[Multigenerational Proteolytic Inactivation of Restriction Upon Subtle Genomic Hypomethylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.22.619740v1?rss=1">
<title>
<![CDATA[
Endogenous neuronal DNA double-strand breaks are not sufficient to drive brain aging and neurodegeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.22.619740v1?rss=1</link>
<description><![CDATA[
Loss of genomic information due to the accumulation of somatic DNA damage has been implicated in aging and neurodegeneration1-3. Somatic mutations in human neurons increase with age4, but it is unclear whether this is a cause or a consequence of brain aging. Here, we clarify the role of endogenous, neuronal DNA double-strand breaks (DSBs) in brain aging and neurodegeneration by generating mice with post-developmental inactivation of the classical non-homologous end-joining (C-NHEJ) core factor Xrcc4 in forebrain neurons. Xrcc4 is critical for the ligation step of C-NHEJ and has no known function outside of DSB repair5,6. We find that, unlike their wild-type counterparts, C-NHEJ-deficient neurons accumulate high levels of DSB foci with age, indicating that neurons undergo frequent DSBs that are typically efficiently repaired by C-NHEJ across their lifespan. Genome-wide mapping reveals that endogenous neuronal DSBs preferentially occur in promoter regions and other genic features. Analysis of 3-D genome organization shows intra-chromosomal clustering and loop extrusion of neuronal DSB regions. Strikingly, however, DSB accumulation caused by loss of C-NHEJ induces only minor epigenetic alterations and does not significantly affect gene expression, 3-D genome organization, or mutational outcomes at neuronal DSBs. Despite extensive aging-associated accumulation of neuronal DSBs, mice with neuronal Xrcc4 inactivation do not show neurodegeneration, neuroinflammation, reduced lifespan, or impaired memory and learning behavior. We conclude that the formation of spontaneous neuronal DSBs caused by normal cellular processes is insufficient to cause brain aging and neurodegeneration, even in the absence of C-NHEJ, the principal neuronal DSB repair pathway.
]]></description>
<dc:creator>Cohen, S.</dc:creator>
<dc:creator>Cheradame, L.</dc:creator>
<dc:creator>Pratt, K.</dc:creator>
<dc:creator>Collins, S.</dc:creator>
<dc:creator>Barillas, A.</dc:creator>
<dc:creator>Carlson, A.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Legube, G.</dc:creator>
<dc:creator>Villeda, S.</dc:creator>
<dc:creator>Mullins, D.</dc:creator>
<dc:creator>Schwer, B.</dc:creator>
<dc:date>2024-10-23</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.619740</dc:identifier>
<dc:title><![CDATA[Endogenous neuronal DNA double-strand breaks are not sufficient to drive brain aging and neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.23.619748v1?rss=1">
<title>
<![CDATA[
Type 2 diabetes remodels collateral circulation and promotes leukocyte adhesion following ischemic stroke 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.23.619748v1?rss=1</link>
<description><![CDATA[
AbstractType 2 diabetes mellitus (T2DM) is associated with impaired leptomeningeal collateral compensation and poor stroke outcome. Neutrophils tethering and rolling on endothelium after stroke can also independently reduce flow velocity. However, the chronology and topological changes in collateral circulation in T2DM is not yet defined. Here, we describe the spatial and temporal blood flow dynamics and vessel remodeling in pial arteries and veins and leukocyte- endothelial adhesion following middle cerebral artery (MCA) stroke using two-photon microscopy in awake control and T2DM mice. Relative to control mice prior to stroke, T2DM mice already exhibited smaller pial vessels with reduced flow velocity. Following stroke, T2DM mice displayed persistently reduced blood flow in pial arteries and veins, resulting in a poor recovery of downstream penetrating arterial flow and a sustained deficit in microvascular flow. There was also persistent increase of leukocyte adhesion to the endothelium of veins, coincided with elevated neutrophils infiltration into brain parenchyma in T2DM mice compared to control mice after stroke. Our data suggest that T2DM-induced increase in chronic inflammation may contribute to the remodeling of leptomeningeal collateral circulation and the observed hemodynamics deficiency that potentiates poor stroke outcome.

HighlightsO_LIBlood flow and leukocyte imaging in awake mice by two-photon microscopy before and after stroke under physiological conditions
C_LIO_LIT2DM induces collateral remodeling prior to stroke
C_LIO_LIT2DM reduces blood flow and impedes recovery in pial arteries and veins after ischemic stroke
C_LIO_LIPoor recovery of penetrating arterial flow and sustained deficit in microvascular flow after ischemic stroke in T2DM mice
C_LIO_LIT2DM increases persistent leukocyte adhesion to endothelium of veins and elevates neutrophils infiltration into the brain parenchyma after ischemic stroke.
C_LI
]]></description>
<dc:creator>Sato, Y.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Kato, Y.</dc:creator>
<dc:creator>Kanoke, A.</dc:creator>
<dc:creator>Sun, Y. J.</dc:creator>
<dc:creator>Nishijima, Y.</dc:creator>
<dc:creator>Wang, R. K.</dc:creator>
<dc:creator>Stryker, M.</dc:creator>
<dc:creator>Endo, H.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:date>2024-10-23</dc:date>
<dc:identifier>doi:10.1101/2024.10.23.619748</dc:identifier>
<dc:title><![CDATA[Type 2 diabetes remodels collateral circulation and promotes leukocyte adhesion following ischemic stroke]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.22.617161v1?rss=1">
<title>
<![CDATA[
Single-Nuclei Analysis of the Unfolded Protein Response (SNUPR): A Novel Method revealing bortezomib resistance mechanisms in Multiple Myeloma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.22.617161v1?rss=1</link>
<description><![CDATA[
AbstractThe unfolded protein response (UPR) is a key stress resistance pathway that has become a key potential target for improving the efficacy of cancer chemotherapy. The UPR involves the activation of three ER-resident stress sensors: PERK, IRE-1 and ATF6 with different signalling outcomes leading to cell death or survival. These cell-fate decisions are difficult to predict and are the result of the complex interaction of PERK, IRE-1 and ATF6 downstream events that have differences in their dynamics and their interplay. These characteristics of the UPR are still poorly defined due to lack of methods to monitor their activation simultaneously at single-cell level. We developed SNUPR (Single Nuclei analysis of the Unfolded Protein Response), an accessible technique that allows the profiling of the three UPR branches in nuclear suspensions by flow cytometry, and applied it to study UPR dynamics in a cancer-specific context. By performing transcriptomic analysis, we found that ER-stress sensor specific gene signatures correlate with patient survival in several blood malignancies, and by using SNUPR, we detected high heterogeneity during UPR activation in vitro in different human cancer cell lines, which could not be have been predicted by the level of expression of the sensors. Our SNUPR analyses further indicate that this heterogeneity is explained by variations in the intensity and duration of ER stress-induced protein synthesis inhibition via PERK, acting as upstream regulator of both the IRE-1/XBP1 and ATF6 dependent transcriptional programs. We extend the relevance of these observations by demonstrating that IRE-1/XBP1s pathway plays a critical role in bortezomib resistance of multiple myeloma cells and patients. We present here SNUPR, that can be used to monitor UPR dynamics with single-cell resolution and identified clinical contexts in which targeting a specific UPR branch could be detrimental or help circumventing chemotherapy resistance.

One Sentence SummarySNUPR method enable single-cell UPR profiling and reveals the role of IRE-1 axis in predicting bortezomib resistance in multiple myeloma.

HighlightsO_LISNUPR allows simultaneous profiling of PERK, IRE-1 and ATF6 activation with single- cell resolution.
C_LIO_LIInhibition of protein synthesis via PERK control the activation levels of the IRE-1/XBP1s and ATF6 pathway.
C_LIO_LIIRE-1 activation and associated transcriptional signatures predict the outcome of patients with multiple myeloma treated with Bortezomib.
C_LIO_LIIRE-1 activity, but not PERK or ATF6, is essential to acquire bortezomib resistance in multiple myeloma cell lines.
C_LI

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=190 HEIGHT=200 SRC="FIGDIR/small/617161v2_ufig1.gif" ALT="Figure 1">
View larger version (41K):
org.highwire.dtl.DTLVardef@297668org.highwire.dtl.DTLVardef@1cab0eorg.highwire.dtl.DTLVardef@2238b4org.highwire.dtl.DTLVardef@b8ebf6_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Arguello, R. J.</dc:creator>
<dc:creator>Pierre, P.</dc:creator>
<dc:creator>Gigan, J. P.</dc:creator>
<dc:creator>Garcia-Gonzalez, P.</dc:creator>
<dc:creator>Galliot, L.</dc:creator>
<dc:creator>Ghaffar, Y.</dc:creator>
<dc:creator>Flores-Santibanez, F.</dc:creator>
<dc:creator>Seillier, E.</dc:creator>
<dc:creator>Lavignolle-Heguy, R.</dc:creator>
<dc:creator>Fischaux, S.</dc:creator>
<dc:creator>Dos Santos, D.</dc:creator>
<dc:creator>Combes, A.</dc:creator>
<dc:creator>Narita, M.</dc:creator>
<dc:creator>Gatti, E.</dc:creator>
<dc:creator>Nal, B.</dc:creator>
<dc:creator>Rocchi, S.</dc:creator>
<dc:creator>Moreaux, J.</dc:creator>
<dc:date>2024-10-25</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.617161</dc:identifier>
<dc:title><![CDATA[Single-Nuclei Analysis of the Unfolded Protein Response (SNUPR): A Novel Method revealing bortezomib resistance mechanisms in Multiple Myeloma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.22.618529v1?rss=1">
<title>
<![CDATA[
Comprehensive structure-function analysis reveals gain- and loss-of-function mechanisms impacting oncogenic KRAS activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.22.618529v1?rss=1</link>
<description><![CDATA[
To dissect variant-function relationships in the KRAS oncoprotein, we performed deep mutational scanning (DMS) screens for both wild-type and KRASG12D mutant alleles. We defined the spectrum of oncogenic potential for nearly all possible KRAS variants, identifying several novel transforming alleles and elucidating a model to describe the frequency of KRAS mutations in human cancer as a function of transforming potential, mutational probability, and tissue-specific mutational signatures. Biochemical and structural analyses of variants identified in a KRASG12D second-site suppressor DMS screen revealed that attenuation of oncogenic KRAS can be mediated by protein instability and conformational rigidity, resulting in reduced binding affinity to effector proteins, such as RAF and PI3-kinases, or reduced SOS-mediated nucleotide exchange activity. These studies define the landscape of single amino acid alterations that modulate the function of KRAS, providing a resource for the clinical interpretation of KRAS variants and elucidating mechanisms of oncogenic KRAS inactivation for therapeutic exploitation.
]]></description>
<dc:creator>Kwon, J.</dc:creator>
<dc:creator>Dilly, J.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Kim, E.</dc:creator>
<dc:creator>Bian, Y.</dc:creator>
<dc:creator>Dharmaiah, S.</dc:creator>
<dc:creator>Tran, T. H.</dc:creator>
<dc:creator>Kapner, K. S.</dc:creator>
<dc:creator>Ly, S. H.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Rabara, D.</dc:creator>
<dc:creator>Waybright, T. J.</dc:creator>
<dc:creator>Giacomelli, A. O.</dc:creator>
<dc:creator>Hong, A. L.</dc:creator>
<dc:creator>Misek, S. L.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Ravi, A.</dc:creator>
<dc:creator>Beroukhim, R.</dc:creator>
<dc:creator>Lemke, C. T.</dc:creator>
<dc:creator>Haigis, K. M.</dc:creator>
<dc:creator>Esposito, D.</dc:creator>
<dc:creator>Root, D. E.</dc:creator>
<dc:creator>Nissley, D. V.</dc:creator>
<dc:creator>Stephen, A. G.</dc:creator>
<dc:creator>McCormick, F.</dc:creator>
<dc:creator>Simanshu, D. K.</dc:creator>
<dc:creator>Hahn, W. C.</dc:creator>
<dc:creator>Aguirre, A. J.</dc:creator>
<dc:date>2024-10-25</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.618529</dc:identifier>
<dc:title><![CDATA[Comprehensive structure-function analysis reveals gain- and loss-of-function mechanisms impacting oncogenic KRAS activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.23.619793v1?rss=1">
<title>
<![CDATA[
Sequence-dependent scale for translocon-mediated insertion of interfacial helices in membranes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.23.619793v1?rss=1</link>
<description><![CDATA[
Biological membranes consist of a lipid bilayer studded with integral and peripheral membrane proteins. Most -helical membrane proteins require protein-conducting insertases known as translocons to assist in their membrane insertion and folding. While the sequence-dependent propensities for a helix to either translocate through the translocon or insert into the membrane have been codified into numerical hydrophobicity scales, the corresponding propensity to partition into the membrane interface remains unraveled. By engineering diagnostic glycosylation sites around test peptide sequences inserted into a host protein, we devised a system that can differentiate between water-soluble, surface-bound, and transmembrane (TM) states of the sequence based on its glycosylation pattern. Using this system, we determined the sequence-dependent propensities for transfer from the translocon to a TM, interfacial or extramembrane space. UMAP analysis of a large collection of TM and water-soluble helices provide useful embeddings for analysis of these propensities and aid in understanding the physical properties and functions of antimicrobial, lytic, and fusogenic peptides.
]]></description>
<dc:creator>Grau, B.</dc:creator>
<dc:creator>Kormos, R.</dc:creator>
<dc:creator>Bano-Polo, M.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Garcia-Murria, M. J.</dc:creator>
<dc:creator>Hajredini, F.</dc:creator>
<dc:creator>Sanchez del Pino, M.</dc:creator>
<dc:creator>Jo, H.</dc:creator>
<dc:creator>Martinez-Gil, L.</dc:creator>
<dc:creator>Von Heijne, G.</dc:creator>
<dc:creator>DeGrado, W.</dc:creator>
<dc:creator>Mingarro, I.</dc:creator>
<dc:date>2024-10-25</dc:date>
<dc:identifier>doi:10.1101/2024.10.23.619793</dc:identifier>
<dc:title><![CDATA[Sequence-dependent scale for translocon-mediated insertion of interfacial helices in membranes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.23.619844v1?rss=1">
<title>
<![CDATA[
AI-readiness for Biomedical Data: Bridge2AI Recommendations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.23.619844v1?rss=1</link>
<description><![CDATA[
Biomedical research is rapidly adopting artificial intelligence (AI). Yet the inherent complexity of biomedical data preparation requires implementing actionable, robust criteria for ethical and explainable AI (XAI) at the "pre-model" stage, encompassing data acquisition, detailed transformations, and ethical governance. Simple conformance to FAIR (Findable, Accessible, Interoperable, Reusable) Principles is insufficient.

Here, we define criteria and practices for reliable AI-readiness of biomedical data, developed by the NIH Bridge to Artificial Intelligence (Bridge2AI) Standards Working Group across seven core dimensions of dataset AI-readiness: FAIRness, Provenance, Characterization, Ethics, Pre-model Explainability, Sustainability, and Computability. Conformance to these criteria provides a basis for pre-model scientific rigor and ethical integrity, mitigating downstream risks of bias and error before AI modeling. We apply and evaluate these standards across all four Bridge2AI flagship datasets, spanning functional genomics to clinical medicine, and encode them in machine-actionable metadata bound to the datasets.

This framework sets a benchmark for preparing ethical, reusable datasets in biomedical AI and provides standardized methods for reliable pre-model data evaluation.
]]></description>
<dc:creator>Clark, T.</dc:creator>
<dc:creator>Caufield, H.</dc:creator>
<dc:creator>Mohan, J. A.</dc:creator>
<dc:creator>Al Manir, S.</dc:creator>
<dc:creator>Amorim, E.</dc:creator>
<dc:creator>Eddy, J.</dc:creator>
<dc:creator>Gim, N.</dc:creator>
<dc:creator>Gow, B.</dc:creator>
<dc:creator>Goar, W.</dc:creator>
<dc:creator>Haendel, M.</dc:creator>
<dc:creator>Hansen, J. N.</dc:creator>
<dc:creator>Harris, N.</dc:creator>
<dc:creator>Hermjakob, H.</dc:creator>
<dc:creator>McWeeney, S. K.</dc:creator>
<dc:creator>Nebeker, C.</dc:creator>
<dc:creator>Nikolov, M.</dc:creator>
<dc:creator>Shaffer, J.</dc:creator>
<dc:creator>Sheffield, N.</dc:creator>
<dc:creator>Sheynkman, G.</dc:creator>
<dc:creator>Stevenson, J.</dc:creator>
<dc:creator>Mungall, C.</dc:creator>
<dc:creator>Chen, J. Y.</dc:creator>
<dc:creator>Wagner, A.</dc:creator>
<dc:creator>Kong, S. W.</dc:creator>
<dc:creator>Ghosh, S. S.</dc:creator>
<dc:creator>Patel, B.</dc:creator>
<dc:creator>Williams, A.</dc:creator>
<dc:creator>Munoz-Torres, M. C.</dc:creator>
<dc:date>2024-10-25</dc:date>
<dc:identifier>doi:10.1101/2024.10.23.619844</dc:identifier>
<dc:title><![CDATA[AI-readiness for Biomedical Data: Bridge2AI Recommendations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.23.619941v1?rss=1">
<title>
<![CDATA[
Imaging synaptic density in ageing and Alzheimer's Disease with -SynVesT-1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.23.619941v1?rss=1</link>
<description><![CDATA[
Monitoring synaptic injury in neurodegenerative diseases may provide new insights into the evolution of the degenerative process as well as a potential mechanism to target for preservation of function. Synaptic density imaging with PET is a relatively new approach to this issue. However, there are remaining questions about technical approaches to data analysis including reference region selection, and how specific phenotypic presentations and symptoms of Alzheimers Disease (AD) are reflected in alterations in synaptic density.

MethodsUsing an SV2A PET ligand radiolabeled with the 18F isotope ([18F]-SynVesT-1) we performed sensitivity analyses to determine the optimal reference tissue modelling approach to derive whole brain ratio images. Using these whole brain images from a sample of young adults, older adults, and patients with varied phenotypic presentations of AD we then contrast regional SV2A density and in vivo AD biomarkers.

ResultReference tissue optimisation concluded that a cerebellar grey matter reference region is best for deriving whole brain ratio images. Using these whole brain ratio images, we find a strong inverse association between [18F]-SynVesT-1 PET uptake and amyloid beta and tau PET deposition. Finally, we find that individuals with lower temporal grey matter volume but higher temporal [18F]-SynVesT-1 PET uptake show preserved performance on the MMSE.

Conclusions[18F]-SynVesT-1 PET shows a close association with in vivo AD pathology and preserved SV2A density may be a possible marker for resilience to neurodegeneration.
]]></description>
<dc:creator>Giorgio, J.</dc:creator>
<dc:creator>Soleimani-Meigooni, D. N.</dc:creator>
<dc:creator>Janabi, M.</dc:creator>
<dc:creator>Baker, S.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Toueg, T. N.</dc:creator>
<dc:creator>Weimer, R.</dc:creator>
<dc:creator>Zinnhardt, B.</dc:creator>
<dc:creator>Green, A.</dc:creator>
<dc:creator>Rabinovici, G. D.</dc:creator>
<dc:creator>Jagust, W.</dc:creator>
<dc:date>2024-10-26</dc:date>
<dc:identifier>doi:10.1101/2024.10.23.619941</dc:identifier>
<dc:title><![CDATA[Imaging synaptic density in ageing and Alzheimer's Disease with -SynVesT-1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.24.620089v1?rss=1">
<title>
<![CDATA[
Encoding manifolds constructed from grating responses organize responses to natural scenes in cortical visual areas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.24.620089v1?rss=1</link>
<description><![CDATA[
A challenge in sensory neuroscience is understanding how populations of neurons operate in concert to represent diverse stimuli. To meet this challenge, we have created "encoding manifolds" that reveal the overall responses of brain areas to diverse stimuli and organize individual neurons according to their selectivity and response dynamics. Here we use encoding manifolds to compare the population-level encoding of primary visual cortex (VISp) with that of five higher visual areas (VISam, VISal, VISpm, VISlm, and VISrl), using data from the Allen Institute Visual Coding-Neuropixels dataset from the mouse. We show that the topology of the encoding manifold for VISp and for higher visual areas is continuous, with smooth coordinates along which stimulus selectivity and response dynamics are organized with layer and cell-type specificity. Surprisingly, the manifolds revealed novel relationships between how natural scenes are encoded relative to static gratings--a relationship conserved across visual areas. Namely, neurons preferring natural scenes preferred either low or high spatial frequency gratings, but not intermediate ones. Analyzing responses by cortical layer reveals a preference for gratings concentrated in layer 6, whereas preferences for natural scenes tended to be higher in layers 2/3 and 4. The results demonstrate how machine learning approaches can be used to organize and visualize the structure of sensory coding, thereby revealing novel relationships within and across brain areas and sensory stimuli.

Significance StatementManifolds have become commonplace for analyzing and visualizing neural responses. However, prior work has focused on building manifolds that organize diverse stimuli in neural response coordinates. Here, we demonstrate the utility of an alternative approach: building manifolds to represent neurons in stimulus/response coordinates, which we term  encoding manifolds. This approach has several advantages, such as being able to directly visualize and compare how different brain areas encode diverse stimulus ensembles. This approach reveals novel relationships between layer-specific responses and the encoding of natural versus artificial stimuli.
]]></description>
<dc:creator>Dyballa, L.</dc:creator>
<dc:creator>Field, G. D.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:creator>Zucker, S. W.</dc:creator>
<dc:date>2024-10-26</dc:date>
<dc:identifier>doi:10.1101/2024.10.24.620089</dc:identifier>
<dc:title><![CDATA[Encoding manifolds constructed from grating responses organize responses to natural scenes in cortical visual areas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.25.620280v1?rss=1">
<title>
<![CDATA[
Differential Sensitivity of Midline Patterning to Mitosis during and after Primitive Streak Extension 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.25.620280v1?rss=1</link>
<description><![CDATA[
BackgroundMidline establishment is a fundamental process during early embryogenesis for Bilaterians. Midline patterning in nonamniotes can occur without mitosis, through Planar Cell Polarity (PCP) signaling. By contrast, amniotes utilize both cell proliferation and PCP signaling for patterning early midline landmark, the primitive streak (PS). This study examined their roles for midline patterning at post PS-extension.

ResultsIn contrast to PS extension stages, embryos under mitotic arrest during the post PS-extension preserved notochord (NC) extension and Hensens node (HN)/PS regression judged by both morphology and marker genes, although they became shorter, and laterality was lost. Remarkably, no or background level of expression was detected for the majority of PCP core components in the NC-HN-PS area at post PS-extension stages, except for robustly detected prickle-1. Morpholino knockdown of Prickle-1 showed little influence on midline patterning, except for suppressed embryonic growth. Lastly, associated with mitotic arrest-induced size reduction, midline tissue cells displayed hypertrophy.

ConclusionThus, the study has identified at least two distinct mitosis sensitivity phases during early midline pattering: One is PS extension that requires both mitosis and PCP, and the other is mitotic arrest-resistant midline patterning with little influence by PCP at post PS-extension stages.
]]></description>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Asai, R.</dc:creator>
<dc:creator>Mikawa, T.</dc:creator>
<dc:date>2024-10-26</dc:date>
<dc:identifier>doi:10.1101/2024.10.25.620280</dc:identifier>
<dc:title><![CDATA[Differential Sensitivity of Midline Patterning to Mitosis during and after Primitive Streak Extension]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.25.620297v1?rss=1">
<title>
<![CDATA[
APOBEC3A drives metastasis of ovarian cancer by altering epithelial-to-mesenchymal transition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.25.620297v1?rss=1</link>
<description><![CDATA[
High-grade serous ovarian cancer (HGSOC) is the most prevalent and aggressive histological subtype of ovarian cancer, and often presents with metastatic disease. The drivers of metastasis in HGSOC remain enigmatic. APOBEC3A (A3A), an enzyme that generates mutations across various cancers, has been proposed as a mediator of tumor heterogeneity and disease progression. However, the role of A3A in HGSOC has not been explored. Through analysis of genome sequencing from primary HGSOC, we observed an association between high levels of APOBEC3 mutagenesis and poor overall survival. We experimentally addressed this correlation by modeling A3A activity in HGSOC cell lines and mouse models which resulted in increased metastatic behavior of HGSOC cells in culture and distant metastatic spread in vivo. A3A activity in both primary and cultured HGSOC cells yielded consistent alterations in expression of epithelial-mesenchymal-transition (EMT) genes resulting in hybrid EMT and mesenchymal signatures, and providing a mechanism for their increased metastatic potential. Our findings define the prevalence of A3A mutagenesis in HGSOC and implicate A3A as a driver of HGSOC metastasis via EMT, underscoring its clinical relevance as a potential prognostic biomarker. Our study lays the groundwork for the development of targeted therapies aimed at mitigating the deleterious impact of A3A-driven EMT in HGSOC.
]]></description>
<dc:creator>Devenport, J. M.</dc:creator>
<dc:creator>Tran, T.</dc:creator>
<dc:creator>Harris, B. R.</dc:creator>
<dc:creator>Fingerman, D. F.</dc:creator>
<dc:creator>DeWeerd, R. A.</dc:creator>
<dc:creator>Elkhidir, L.</dc:creator>
<dc:creator>LaVigne, D. A.</dc:creator>
<dc:creator>Fuh, K.</dc:creator>
<dc:creator>Sun, L.</dc:creator>
<dc:creator>Bednarski, J. J.</dc:creator>
<dc:creator>Drapkin, R.</dc:creator>
<dc:creator>Mullen, M.</dc:creator>
<dc:creator>Green, A. M.</dc:creator>
<dc:date>2024-10-28</dc:date>
<dc:identifier>doi:10.1101/2024.10.25.620297</dc:identifier>
<dc:title><![CDATA[APOBEC3A drives metastasis of ovarian cancer by altering epithelial-to-mesenchymal transition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.27.620437v1?rss=1">
<title>
<![CDATA[
Restoring the Multiple Sclerosis Associated Imbalance of Gut Indole Metabolites Promotes Remyelination and Suppresses Neuroinflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.27.620437v1?rss=1</link>
<description><![CDATA[
In multiple sclerosis (MS) the circulating metabolome is dysregulated, with indole lactate (ILA) being one of the most significantly reduced metabolites. We demonstrate that oral supplementation of ILA impacts key MS disease processes in two preclinical models. ILA reduces neuroinflammation by dampening immune cell activation as well as infiltration; and promotes remyelination and in vitro oligodendrocyte differentiation through the aryl hydrocarbon receptor (AhR). Supplementation of ILA, a reductive indole metabolite, restores the gut microbiomes oxidative/reductive metabolic balance by lowering circulating indole acetate (IAA), an oxidative indole metabolite, that blocks remyelination and oligodendrocyte maturation. The ILA-induced reduction in circulating IAA is linked to changes in IAA-producing gut microbiota taxa and pathways that are also dysregulated in MS. Notably, a lower ILA:IAA ratio correlates with worse MS outcomes. Overall, these findings identify ILA as a potential anti-inflammatory remyelinating agent and provide insights into the role of gut dysbiosis-related metabolic alterations in MS progression.

One Sentence SummaryIndole lactate, a postbiotic metabolite reduced in MS, corrects gut microbiome metabolic imbalances associated with remyelination and neuroinflammation.
]]></description>
<dc:creator>Jank, L.</dc:creator>
<dc:creator>Singh, S. S.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Dhukhwa, A.</dc:creator>
<dc:creator>Siavoshi, F.</dc:creator>
<dc:creator>Joshi, D.</dc:creator>
<dc:creator>Minney, V.</dc:creator>
<dc:creator>Gupta, K.</dc:creator>
<dc:creator>Ghimire, S.</dc:creator>
<dc:creator>Deme, P.</dc:creator>
<dc:creator>Schoeps, V. A.</dc:creator>
<dc:creator>Soman, K.</dc:creator>
<dc:creator>Ladakis, D.</dc:creator>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Borkowski, K.</dc:creator>
<dc:creator>Newman, J.</dc:creator>
<dc:creator>Baranzini, S. E.</dc:creator>
<dc:creator>Waubant, E. L.</dc:creator>
<dc:creator>Fitzgerald, K. C.</dc:creator>
<dc:creator>Mangalam, A. K.</dc:creator>
<dc:creator>Haughey, N.</dc:creator>
<dc:creator>Kornberg, M. D.</dc:creator>
<dc:creator>Chamling, X.</dc:creator>
<dc:creator>Calabresi, P. A.</dc:creator>
<dc:creator>Bhargava, P.</dc:creator>
<dc:date>2024-10-28</dc:date>
<dc:identifier>doi:10.1101/2024.10.27.620437</dc:identifier>
<dc:title><![CDATA[Restoring the Multiple Sclerosis Associated Imbalance of Gut Indole Metabolites Promotes Remyelination and Suppresses Neuroinflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.25.620326v1?rss=1">
<title>
<![CDATA[
Phosphorylation of HP1/Swi6 relieves competition with Suv39/Clr4 on nucleosomes and enables H3K9 trimethyl spreading. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.25.620326v1?rss=1</link>
<description><![CDATA[
Heterochromatin formation in Schizosaccharomyces pombe requires the spreading of histone 3 (H3) Lysine 9 (K9) methylation (me) from nucleation centers by the H3K9 methylase, Suv39/Clr4, and the reader protein, HP1/Swi6. To accomplish this, Suv39/Clr4 and HP1/Swi6 have to associate with nucleosomes both nonspecifically, binding DNA and octamer surfaces and specifically, via recognition of methylated H3K9 by their respective chromodomains. However, how both proteins avoid competition for the same nucleosomes in this process is unclear. Here, we show that phosphorylation tunes oligomerization and the nucleosome affinity of HP1/Swi6 such that it preferentially partitions onto Suv39/Clr4s trimethyl product rather than its unmethylated substrates. Preferential partitioning enables efficient conversion from di-to trimethylation on nucleosomes in vitro and H3K9me3 spreading in vivo. Together, our data suggests that phosphorylation of HP1/Swi6 creates a regime that increases oligomerization and relieves competition with the "read-write" mechanism of Suv39/Clr4, together promoting for productive heterochromatin spreading.
]]></description>
<dc:creator>Kennedy, D. R.</dc:creator>
<dc:creator>Lemiere, J.</dc:creator>
<dc:creator>Tan, C.</dc:creator>
<dc:creator>Simental, E.</dc:creator>
<dc:creator>Braxton, J.</dc:creator>
<dc:creator>Maxwell, R. A.</dc:creator>
<dc:creator>Amine, A. A.</dc:creator>
<dc:creator>Al-Sady, B.</dc:creator>
<dc:date>2024-10-29</dc:date>
<dc:identifier>doi:10.1101/2024.10.25.620326</dc:identifier>
<dc:title><![CDATA[Phosphorylation of HP1/Swi6 relieves competition with Suv39/Clr4 on nucleosomes and enables H3K9 trimethyl spreading.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.26.620318v1?rss=1">
<title>
<![CDATA[
DNMT3AR882H Is Not Required for Disease Maintenance in Primary Human AML, but Is Associated With Increased Leukemia Stem Cell Frequency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.26.620318v1?rss=1</link>
<description><![CDATA[
Genetic mutations are being thoroughly mapped in human cancers, yet a fundamental question in cancer biology is whether such mutations are functionally required for cancer initiation, maintenance of established cancer, or both. Here, we study this question in the context of human acute myeloid leukemia (AML), where DNMT3AR882 missense mutations often arise early, in pre-leukemic clonal hematopoiesis, and corrupt the DNA methylation landscape to initiate leukemia. We developed CRISPR-based methods to directly correct DNMT3AR882 mutations in leukemic cells obtained from patients. Surprisingly, DNMT3AR882 mutations were largely dispensable for disease maintenance. Replacing DNMT3AR882 mutants with wild-type DNMT3A did not impair the ability of AML cells to engraft in vivo, and minimally altered DNA methylation. Taken together, DNMT3AR882 mutations are initially necessary for AML initiation, but are largely dispensable for disease maintenance. The notion that initiating oncogenes differ from those that maintain cancer has important implications for cancer evolution and therapy.

STATEMENT OF SIGNIFICANCEUnderstanding which driver mutations are required for cancer initiation, maintenance, or both phases remains poorly understood. Here, we uncover that highly prevalent pre-leukemic DNMT3A mutations are only required during disease initiation, but become dispensable after leukemic transformation, uncovering the context-specific role of this driver mutation with important therapeutic implications.
]]></description>
<dc:creator>Koehnke, T.</dc:creator>
<dc:creator>Karigane, D.</dc:creator>
<dc:creator>Hilgart, E.</dc:creator>
<dc:creator>Fan, A. C.</dc:creator>
<dc:creator>Kayamori, K.</dc:creator>
<dc:creator>Miyauchi, M.</dc:creator>
<dc:creator>Collins, C. T.</dc:creator>
<dc:creator>Suchy, F. P.</dc:creator>
<dc:creator>Rangavajhula, A.</dc:creator>
<dc:creator>Feng, Y.</dc:creator>
<dc:creator>Nakauchi, Y.</dc:creator>
<dc:creator>Martinez-Montes, E.</dc:creator>
<dc:creator>Fowler, J.</dc:creator>
<dc:creator>Loh, K. M.</dc:creator>
<dc:creator>Nakauchi, H.</dc:creator>
<dc:creator>Koldobskiy, M. A.</dc:creator>
<dc:creator>Feinberg, A. P.</dc:creator>
<dc:creator>Majeti, R.</dc:creator>
<dc:date>2024-10-29</dc:date>
<dc:identifier>doi:10.1101/2024.10.26.620318</dc:identifier>
<dc:title><![CDATA[DNMT3AR882H Is Not Required for Disease Maintenance in Primary Human AML, but Is Associated With Increased Leukemia Stem Cell Frequency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.27.620516v1?rss=1">
<title>
<![CDATA[
A multi-subunit autophagic capture complex facilitates degradation of ER stalled MHC-I in pancreatic cancer. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.27.620516v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDA) evades immune detection partly via autophagic capture and lysosomal degradation of major histocompatibility complex class I (MHC-I). Why MHC-I is susceptible to capture via autophagy remains unclear. By synchronizing exit of proteins from the endoplasmic reticulum (ER), we show that PDAC cells display prolonged retention of MHC-I in the ER and fail to efficiently route it to the plasma membrane. A capture-complex composed of NBR1 and the ER-phagy receptor TEX264 facilitates targeting of MHC-I for autophagic degradation, and suppression of either receptor is sufficient to increase total levels and re-route MHC-I to the plasma membrane. Binding of MHC-I to the capture complex is linked to antigen presentation efficiency, as inhibiting antigen loading via knockdown of TAP1 or beta 2-Microglobulin led to increased binding between MHC-I and the TEX264-NBR1 capture complex. Conversely, expression of ER directed high affinity antigenic peptides led to increased MHC-I at the cell surface and reduced lysosomal degradation. A genome-wide CRISPRi screen identified NFXL1, as an ER-resident E3 ligase that binds to MHC-I and mediates its autophagic capture. High levels of NFXL1 are negatively correlated with MHC-I protein expression and predicts poor patient prognosis. These data highlight an ER resident capture complex tasked with sequestration and degradation of non-conformational MHC-I in PDAC cells, and targeting this complex has the potential to increase PDAC immunogenicity.
]]></description>
<dc:creator>Berquez, M.</dc:creator>
<dc:creator>Li, A. L.</dc:creator>
<dc:creator>Luy, M. A.</dc:creator>
<dc:creator>Venida, A. C.</dc:creator>
<dc:creator>O'Loughlin, T.</dc:creator>
<dc:creator>Rademaker, G.</dc:creator>
<dc:creator>Barpanda, A.</dc:creator>
<dc:creator>Hu, J.</dc:creator>
<dc:creator>Yano, J.</dc:creator>
<dc:creator>Wiita, A.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:creator>Bruno, P. M.</dc:creator>
<dc:creator>Perera, R. M.</dc:creator>
<dc:date>2024-10-29</dc:date>
<dc:identifier>doi:10.1101/2024.10.27.620516</dc:identifier>
<dc:title><![CDATA[A multi-subunit autophagic capture complex facilitates degradation of ER stalled MHC-I in pancreatic cancer.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.28.620733v1?rss=1">
<title>
<![CDATA[
A mitochondrial redox switch licenses the onset of morphogenesis in animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.28.620733v1?rss=1</link>
<description><![CDATA[
Embryos undergo pre-gastrulation cleavage cycles to generate a critical cell mass before transitioning to morphogenesis. The molecular underpinnings of this transition have traditionally centered on zygotic chromatin remodeling and genome activation1,2, as their repression can prevent downstream processes of differentiation and organogenesis. Despite precedents that oxygen depletion can similarly suspend development in early embryos3-6, hinting at a pivotal role for oxygen metabolism in this transition, whether there is a bona fide chemical switch that licenses the onset of morphogenesis remains unknown. Here we discover that a mitochondrial oxidant acts as a metabolic switch to license the onset of animal morphogenesis. Concomitant with the instatement of mitochondrial membrane potential, we found a burst-like accumulation of mitochondrial superoxide (O2-) during fly blastoderm formation. In vivo chemistry experiments revealed that an electron leak from site IIIQo at ETC Complex III is responsible for O2- production. Importantly, depleting mitochondrial O2- fully mimics anoxic conditions and, like anoxia, induces suspended animation prior to morphogenesis, but not after. Specifically, H2O2, and not ONOO-, NO, or HO*, can single-handedly account for this mtROS-based response. We demonstrate that depleting mitochondrial O2- similarly prevents the onset of morphogenetic events in vertebrate embryos and ichthyosporea, close relatives of animals. We postulate that such redox-based metabolic licensing of morphogenesis is an ancient trait of holozoans that couples the availability of oxygen to development, conserved from early-diverging animal relatives to vertebrates.
]]></description>
<dc:creator>Kahlin, U.</dc:creator>
<dc:creator>Dalla Ricca, F.</dc:creator>
<dc:creator>Pillai, S. J.</dc:creator>
<dc:creator>Olivetta, M.</dc:creator>
<dc:creator>Tharp, K. M.</dc:creator>
<dc:creator>Jao, L.-E.</dc:creator>
<dc:creator>Dudin, O.</dc:creator>
<dc:creator>McDonald, K. L.</dc:creator>
<dc:creator>Aydogan, M. G.</dc:creator>
<dc:date>2024-10-29</dc:date>
<dc:identifier>doi:10.1101/2024.10.28.620733</dc:identifier>
<dc:title><![CDATA[A mitochondrial redox switch licenses the onset of morphogenesis in animals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.29.620718v1?rss=1">
<title>
<![CDATA[
eIF4ET regulates meiotic proteome levels to enable oocyte formation and storage 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.29.620718v1?rss=1</link>
<description><![CDATA[
Animals store oocytes in a dormant state for weeks to decades before ovulation. The homeostatic programs oocytes use to endure long-term storage are poorly understood. Using female nematodes as a short-lived model, we found that oocyte formation and storage required IFET-1, the conserved eIF4E-Transporter protein (eIF4ET). IFET-1 co-assembled with CAR-1 (Lsm14) to form micron-scale condensates in stored oocytes, which dissipated after oocyte activation. Depletion of IFET-1 destabilized the stored oocyte proteome, leading to lower translation, a decline in microtubule maintenance proteins, and errors in microtubule organization and meiotic spindle assembly. Deleting individual domains within IFET-1 impaired oocyte storage without affecting oocyte formation. Thus, in addition to establishing a healthy oocyte reserve in young mothers, IFET-1 ensures that correct levels of cytoskeletal proteins are maintained as oocytes age. Human eIF4ET also localized to micron-scale puncta in dormant oocytes in a reproductively healthy patient. Our results clarify the role for eIF4ET in maintaining the oocyte reserve and further support eIF4ET dysfunction as an upstream cause of embryonic aneuploidy and age-related infertility.
]]></description>
<dc:creator>Bhatia, P.</dc:creator>
<dc:creator>Amin, R.</dc:creator>
<dc:creator>Familiari, N. E.</dc:creator>
<dc:creator>Yaguchi, K.</dc:creator>
<dc:creator>Tran, V. M.</dc:creator>
<dc:creator>Bond, A.</dc:creator>
<dc:creator>Bukulmez, O.</dc:creator>
<dc:creator>Woodruff, J. B.</dc:creator>
<dc:date>2024-10-29</dc:date>
<dc:identifier>doi:10.1101/2024.10.29.620718</dc:identifier>
<dc:title><![CDATA[eIF4ET regulates meiotic proteome levels to enable oocyte formation and storage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.29.620973v1?rss=1">
<title>
<![CDATA[
Orexinergic projections to substantia innominata mediate arousal and analgesia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.29.620973v1?rss=1</link>
<description><![CDATA[
Understanding neural circuits involved in anesthesia is crucial for improving its safety and efficacy. Hypothalamic orexin neurons (LHAOX), projecting broadly, are essential in regulating arousal and pain. However, the precise targets remain unclear. Here we investigated the orexin projections to the substantia innominata (SI). Combining optogenetics, fiber photometry, and EEG/EMG allowed us to manipulate orexin activities, while simultaneously recording local ligand release and global cortical activities during anesthesia. Brain slice electrophysiology revealed the synaptic connections in the SI, while RNAscope was employed to examine the distribution of orexin receptors and downstream neuronal types. Presynaptic vesicles were identified in the orexin terminals in the SI, where 49.16% of cells expressed OX2R and 6.8% expressed OX1R. Orexin release in the SI was reversibly suppressed by isoflurane. Optogenetic activation of the LHAOX[-&gt;]SI circuit significantly increased orexin release and promoted arousal from various anesthesia stages, including reanimation during 0.75% isoflurane (p < 0.0001), prolongation of 3% isoflurane induction (p = 0.0033), and acceleration of emergence from 2% isoflurane (p < 0.0001). Furthermore, activating this circuit induced analgesia to both thermal (p = 0.0074) and inflammatory (p = 0.0127) pain. Patch-clamp recordings revealed that optogenetic activation of orexin terminals in the SI elicited excitatory postsynaptic currents, which were blocked by the OX2R antagonist. SI contains more GABAergic (28.17%) and glutamatergic (11.96%) neurons than cholinergic neurons (4.13%), all of which expressed OX2R. Thus, LHAOX neurons innervate SI neurons to regulate both arousal and pain predominantly through OX2R.
]]></description>
<dc:creator>Xiang, X.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Guan, Z.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:date>2024-10-31</dc:date>
<dc:identifier>doi:10.1101/2024.10.29.620973</dc:identifier>
<dc:title><![CDATA[Orexinergic projections to substantia innominata mediate arousal and analgesia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.30.621078v1?rss=1">
<title>
<![CDATA[
A Novel Human TBCK- Neuronal Cell Model Results in Severe Neurodegeneration and Partial Rescue with Mitochondrial Fission Inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.30.621078v1?rss=1</link>
<description><![CDATA[
Background and ObjectivesTBCK syndrome is a rare fatal pediatric neurodegenerative disease caused by biallelic loss-of-function mutations in the TBCK gene. Previous studies by our lab and others have implicated mTOR, autophagy, lysosomes, and intracellular mRNA transport, however the exact primary pathologic mechanism is unknown. This gap has prevented the development of targeted therapies.

MethodsWe employed a human neural progenitor cell line (NPC), ReNcell VM, which can differentiate into neurons and astrocytes, to understand the role of TBCK in mTORC1 activity and neuronal autophagy and cellular mechanisms of pathology. We used shRNA technology to knockdown TBCK in ReNcells.

ResultsThese data showed that loss of TBCK did not inhibit mTORC1 activity in neither NPC nor neurons. Additionally, analysis of eight patient-derived cells and TBCK knock down HeLa cells showed that mTORC1 inhibition is inconsistent across different patients and cell types. We showed that TBCK knockdown in ReNcells affected NPC differentiation to neurons and astrocytes. Specifically, differentiation defects are coupled to cell cycle defects in NPC and increased cell death during differentiation. RNAseq analysis indicated the downregulation of several different neurodevelopmental and differentiation pathways. We observed a higher number of LC3-positive vesicles in the soma and neurites of TBCK knockdown cells. Further, TBCK knockdown altered mitochondrial dynamics and membrane potential in NPC, neurons and astrocytes. We found partial mitochondrial rescue with the mitochondrial fission inhibitor mdivi- 1.

DiscussionThis work outlines a new Human Cell Model for TBCK-related neurodegeneration and the essential role of mitochondrial health and partial rescue with mitochondrial fission inhibitor. This data, along with human neurons and astrocytes, illuminate mechanisms of neurodegeneration and provide a possible novel therapeutic avenue for affected patients.
]]></description>
<dc:creator>Angireddy, R.</dc:creator>
<dc:creator>Murali, S.</dc:creator>
<dc:creator>Karisetty, B. C.</dc:creator>
<dc:creator>Kossenkov, A.</dc:creator>
<dc:creator>Katsura, K.</dc:creator>
<dc:creator>Ohl, L.</dc:creator>
<dc:creator>Smith, S.</dc:creator>
<dc:creator>Clark, K.</dc:creator>
<dc:creator>Bhoj, E.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:date>2024-10-31</dc:date>
<dc:identifier>doi:10.1101/2024.10.30.621078</dc:identifier>
<dc:title><![CDATA[A Novel Human TBCK- Neuronal Cell Model Results in Severe Neurodegeneration and Partial Rescue with Mitochondrial Fission Inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.30.621206v1?rss=1">
<title>
<![CDATA[
A designed Zn2+ sensor domain transmits binding information to transmembrane histidine kinases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.30.621206v1?rss=1</link>
<description><![CDATA[
Generating stimulus-responsive, allosteric signaling de novo is a significant challenge in protein design. In natural systems like bacterial histidine kinases (HKs), signal transduction occurs when ligand binding initiates a signal that is amplified across biological membranes over long distances to induce large-scale rearrangements and phosphorylation relays. Here, we ask whether our understanding of protein design and multi-domain, intramolecular signaling has progressed sufficiently to enable engineering of a HK with tunable de novo components. We generated de novo metal-binding sensor domains and substituted them for the native sensor domain of a transmembrane HK, affording chimeras that transduce signals initiated from a de novo sensor. Signaling depended on the designed sensors stability and the interdomain linkers phase and length. These results show the usefulness of de novo design to elucidate biochemical mechanisms and principles for design of new signaling systems.
]]></description>
<dc:creator>Hatstat, A. K.</dc:creator>
<dc:creator>Kormos, R.</dc:creator>
<dc:creator>Xu, V.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:date>2024-10-31</dc:date>
<dc:identifier>doi:10.1101/2024.10.30.621206</dc:identifier>
<dc:title><![CDATA[A designed Zn2+ sensor domain transmits binding information to transmembrane histidine kinases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.31.621371v1?rss=1">
<title>
<![CDATA[
Automated segmentation of soft X-ray tomography: native cellular structure with sub-micron resolution at high throughput for whole-cell quantitative imaging in yeast 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.31.621371v1?rss=1</link>
<description><![CDATA[
Soft X-ray tomography (SXT) is an invaluable tool for quantitatively analyzing cellular structures at sub-optical isotropic resolution. However, it has traditionally depended on manual segmentation, limiting its scalability for large datasets. Here, we leverage a deep learning-based auto-segmentation pipeline to segment and label cellular structures in hundreds of cells across three Saccharomyces cerevisiae strains. This task-based pipeline employs manual iterative refinement to improve segmentation accuracy for key structures, including the cell body, nucleus, vacuole, and lipid droplets, enabling high-throughput and precise phenotypic analysis. Using this approach, we quantitatively compared the 3D whole-cell morphometric characteristics of wild-type, VPH1-GFP, and vac14 strains, uncovering detailed strain-specific cell and organelle size and shape variations. We show the utility of SXT data for precise 3D curvature analysis of entire organelles and cells and detection of fine morphological features using surface meshes. Our approach facilitates comparative analyses with high spatial precision and statistical throughput, uncovering subtle morphological features at the single cell and population level. This workflow significantly enhances our ability to characterize cell anatomy and supports scalable studies on the mesoscale, with applications in investigating cellular architecture, organelle biology, and genetic research across diverse biological contexts.

Significance StatementO_LISoft X-ray tomography offers many powerful features for whole-cell multi-organelle imaging, but, like other high resolution volumetric imaging modalities, is typically limited by low throughput due to laborious segmentation.
C_LIO_LIAuto-segmentation for soft X-ray tomography overcomes this limitation, enabling statistical 3D morphometric analysis of multiple organelles in whole cells across cell populations.
C_LIO_LIThe combination of high 3D resolution of SXT data with statistically useful throughput represents an avenue for more thorough characterizations of cells in toto and opens new mesoscale biological questions and statistical whole-cell modeling of organelle and cell morphology, interactions, and responses to perturbations.
C_LI
]]></description>
<dc:creator>Chen, J.-H. F.</dc:creator>
<dc:creator>Mirvis, M.</dc:creator>
<dc:creator>Ekman, A.</dc:creator>
<dc:creator>Vanslembrouck, B.</dc:creator>
<dc:creator>Le Gros, M.</dc:creator>
<dc:creator>Larabell, C.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2024-11-01</dc:date>
<dc:identifier>doi:10.1101/2024.10.31.621371</dc:identifier>
<dc:title><![CDATA[Automated segmentation of soft X-ray tomography: native cellular structure with sub-micron resolution at high throughput for whole-cell quantitative imaging in yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.01.621612v1?rss=1">
<title>
<![CDATA[
Multi-modal characterization of rodent tooth development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.01.621612v1?rss=1</link>
<description><![CDATA[
Craniofacial tissues undergo hard tissue development through mineralization and changes in physicochemical properties. This study investigates the mechanical and chemical properties of developing enamel, dentin, and bone in the mouse mandible. We employ a multi-modal, multi-scale analysis of the developing incisor and first molar at postnatal day 12 by integrating micro-computed tomography (microCT), nanoindentation (NI), energy dispersive spectroscopy (EDS), and Raman spectroscopy. Our findings demonstrate distinct patterns of mechanical, elemental, and chemical changes across mineralized tissues. These results suggest that mineral composition drives mechanical properties across different craniofacial hard tissues. Integrating multi-modal characterization of mineralized tissues opens new opportunities for investigating structure-function relationships in craniofacial biology and genetics.
]]></description>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Katsura, K. A.</dc:creator>
<dc:creator>Badt, N. Z.</dc:creator>
<dc:creator>Didziokas, M.</dc:creator>
<dc:creator>Dougherty, S.</dc:creator>
<dc:creator>Goldsby, D. L.</dc:creator>
<dc:creator>Bhoj, E. J.</dc:creator>
<dc:creator>Vining, K. H.</dc:creator>
<dc:date>2024-11-01</dc:date>
<dc:identifier>doi:10.1101/2024.11.01.621612</dc:identifier>
<dc:title><![CDATA[Multi-modal characterization of rodent tooth development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.01.621613v1?rss=1">
<title>
<![CDATA[
Heart tube morphogenesis is regulated by segment-specific gene regulatory networks controlled by MEF2C 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.01.621613v1?rss=1</link>
<description><![CDATA[
The gene regulatory networks (GRNs) that control early heart formation are beginning to be understood, but lineage-specific GRNs remain largely undefined. We investigated networks controlled by the vital transcription factor MEF2C, with a time course of single-nucleus RNA- and ATAC-sequencing in wild-type and Mef2c-null embryos. We identified a "posteriorized" cardiac gene signature and chromatin landscape in the absence of MEF2C. Integrating our multiomics data in a deep learning-based model, we constructed developmental trajectories for each of the outflow tract, ventricular, and inflow tract segments, and alterations of these in Mef2c-null embryos. We computationally identified segment-specific MEF2C-dependent enhancers, with activity in the developing zebrafish heart. Finally, using inferred GRNs we discovered that the Mef2c-null heart malformations are partly driven by increased activity of the nuclear hormone receptor NR2F2. Our results delineate lineage-specific GRNs in the early heart tube and provide a generalizable framework for dissecting transcriptional networks governing developmental processes.
]]></description>
<dc:creator>Muncie-Vasic, J. M.</dc:creator>
<dc:creator>Sinha, T.</dc:creator>
<dc:creator>Clark, A. P.</dc:creator>
<dc:creator>Brower, E. F.</dc:creator>
<dc:creator>Saucerman, J. J.</dc:creator>
<dc:creator>Black, B. L.</dc:creator>
<dc:creator>Bruneau, B. G.</dc:creator>
<dc:date>2024-11-01</dc:date>
<dc:identifier>doi:10.1101/2024.11.01.621613</dc:identifier>
<dc:title><![CDATA[Heart tube morphogenesis is regulated by segment-specific gene regulatory networks controlled by MEF2C]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.30.621099v1?rss=1">
<title>
<![CDATA[
Hypoxia induces extensive protein and proteolytic remodeling of the cell surface in pancreatic adenocarcinoma (PDAC) cell lines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.30.621099v1?rss=1</link>
<description><![CDATA[
The tumor microenvironment (TME) plays a crucial role in cancer progression. Hypoxia is a hallmark of the TME and induces a cascade of molecular events that affects cellular processes involved in metabolism, metastasis, and proteolysis. In pancreatic ductal adenocarcinoma (PDAC), tumor tissues are extremely hypoxic. Here, we leveraged mass spectrometry technologies to examine hypoxia-induced alterations in the abundance and proteolytic modifications to cell surface and secreted proteins. Across four PDAC cell lines, we discovered extensive proteolytic remodeling of cell surface proteins involved in cellular adhesion and motility. Looking outward at the surrounding secreted space, we identified hypoxia-regulated secreted and proteolytically-shed proteins that are responsible for regulating the humoral immune and inflammatory response and an upregulation of proteins involved in metabolic processing and tissue development. Combining cell surface N-terminomics and secretomics to evaluate the cellular response to hypoxia enabled us to identify significantly altered candidate proteins which may serve as potential biomarkers and therapeutic targets in PDAC. Furthermore, this approach provides a blue print for studying dysregulated extracellular proteolysis in other cancers and inflammatory diseases.
]]></description>
<dc:creator>Lui, I.</dc:creator>
<dc:creator>Schaefer, K.</dc:creator>
<dc:creator>Kirkemo, L.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Perera, R. M.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2024-11-02</dc:date>
<dc:identifier>doi:10.1101/2024.10.30.621099</dc:identifier>
<dc:title><![CDATA[Hypoxia induces extensive protein and proteolytic remodeling of the cell surface in pancreatic adenocarcinoma (PDAC) cell lines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.01.621560v1?rss=1">
<title>
<![CDATA[
Site-specific DNA insertion into the human genome with engineered recombinases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.01.621560v1?rss=1</link>
<description><![CDATA[
Technologies for precisely inserting large DNA sequences into the genome are critical for diverse research and therapeutic applications. Large serine recombinases (LSRs) can mediate direct, site-specific genomic integration of multi-kilobase DNA sequences without a pre-installed landing pad, but current approaches suffer from low insertion rates and high off-target activity. Here, we present a comprehensive engineering roadmap for the joint optimization of DNA recombination efficiency and specificity. We combined directed evolution, structural analysis, and computational models to rapidly identify additive mutational combinations. We further enhanced performance through donor DNA optimization and dCas9 fusions, enabling simultaneous target and donor recruitment. Top engineered LSR variants achieved up to 53% integration efficiency and 97% genome-wide specificity at an endogenous human locus, and effectively integrated large DNA cargoes (up to 12 kb tested) for stable expression in challenging cell types, including non-dividing cells, human embryonic stem cells, and primary human T cells. This blueprint for rational engineering of DNA recombinases enables precise genome engineering without the generation of double-stranded breaks.
]]></description>
<dc:creator>Fanton, A.</dc:creator>
<dc:creator>Bartie, L. J.</dc:creator>
<dc:creator>Martins, J. Q.</dc:creator>
<dc:creator>Tran, V. Q.</dc:creator>
<dc:creator>Goudy, L.</dc:creator>
<dc:creator>Durrant, M. G.</dc:creator>
<dc:creator>Wei, J.</dc:creator>
<dc:creator>Pawluk, A.</dc:creator>
<dc:creator>Konermann, S.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:creator>Hsu, P. D.</dc:creator>
<dc:date>2024-11-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.01.621560</dc:identifier>
<dc:title><![CDATA[Site-specific DNA insertion into the human genome with engineered recombinases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.04.621823v1?rss=1">
<title>
<![CDATA[
Chromatin mimicry by human JC virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.04.621823v1?rss=1</link>
<description><![CDATA[
Chronically persistent viruses are integral components of the organismal ecosystem in humans and animals 1 2. Many of these viruses replicate and accumulate within the cell nucleus 3. The nuclear location allows viruses to evade cytoplasmic host viral sensors and promotes viral replication 4. One of the unexplored and puzzling aspects of the viral nuclear lifecycle involves the viruss ability to deal with the physical constraints of nuclear architecture. To replicate and accumulate within the nucleus in large numbers sufficient for infection spreading, DNA viruses need to overcome the spatial limitations imposed by chromatin and the nuclear matrix. We found that one of the most widespread and potentially lethal human viruses, the JC polyomavirus 5, interferes with nuclear heterochromatin to create virus-occupied space. The JC viruss impact on heterochromatin is mediated by the viral nonstructural protein, Agnoprotein (Agno). Agnos interference with heterochromatin is governed by structurally diverse mimics of host epigenetic regulators that facilitate virus-induced chromatin reorganization and a dramatic decline in nuclear stiffness in the infected cells. The JCV epigenetic mimicry is critical for the virus infection, as evident from reduced replication of mimic-mutant viruses. Our data suggest that modulation of nuclear mechanical properties is a novel strategy enabling chronicity of the JC and possibly other nuclear virus infections.
]]></description>
<dc:creator>Schaefer, U.</dc:creator>
<dc:creator>Miroshnikova, Y. A.</dc:creator>
<dc:creator>Xie, W.</dc:creator>
<dc:creator>Larson, A. G.</dc:creator>
<dc:creator>Lu, Z.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Bradic, M.</dc:creator>
<dc:creator>Goldgur, Y.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Sharma, V. P.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Patel, D. J.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:creator>Wickstrom, S. A.</dc:creator>
<dc:creator>Tarakhovsky, A.</dc:creator>
<dc:date>2024-11-04</dc:date>
<dc:identifier>doi:10.1101/2024.11.04.621823</dc:identifier>
<dc:title><![CDATA[Chromatin mimicry by human JC virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.04.621904v1?rss=1">
<title>
<![CDATA[
Antigen-Specific T Cell Receptor Discovery for Treating Progressive Multifocal Leukoencephalopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.04.621904v1?rss=1</link>
<description><![CDATA[
BackgroundProgressive multifocal leukoencephalopathy (PML) is a frequently fatal disease of the central nervous system caused by JC virus (JCV). Survival is dependent on early diagnosis and ability to re-establish anti-viral T cell immunity. Adoptive transfer of polyomavirus-specific T cells has shown promise; however, there are no readily available HLA-matched anti-viral T cells to facilitate rapid treatment.

ObjectiveIdentify epitopes of the JCV major capsid protein VP1 that elicit an immune response in the context of human leukocyte antigen allele A*02:01 (HLA-A2) and isolate cognate T cell receptors (TCRs) from healthy donors. Evaluate individual VP1-specific TCRs for their capacity to be expressed in T cells and clear JCV in vitro.

MethodsPBMCs from HLA-A2+ healthy donors were stimulated with peptide libraries tiled across the JCV VP1 protein. Multiple rounds of stimulation were performed to identify the antigens that induced the largest expansion and CD8+ T cell response (measured as INF{gamma}, TNF, CD137, and CD69 expression). High-affinity, antigen-specific CD8+ T cells were isolated based on intensity of tetramer binding for downstream single-cell TCR sequencing. Candidate TCRs were selected based on tetramer binding affinity and activation assays. Promising TCRs were introduced into the T cell genome via viral transduction for in vitro validation including peptide-pulsed K562 cells and astrocyte cells, and JCV-infected astrocytes.

ResultsFour conserved JCV VP1 epitopes (amino acids 100-108, 251-259, 253-262, and 274-283) presented by HLA-A2 were identified. VP1(100-108) consistently elicited the highest level of IFN-{gamma} production from multiple donors and this peptide is in a highly conserved region of VP1. We next identified fourteen high avidity TCRs specific for VP1(100-108). When virally transduced into primary human T cells, seven of these TCRs demonstrated specific binding to VP1(100-108):HLA-A2 tetramers, and four showed increased IFN-{gamma} response when incubated with peptide. Primary CD8+ T cells expressing two of these TCRs cleared both HLA-A2 positive K562 cells and HLA-A2 positive SVG astrocyte cell line presenting exogenously added VP1 peptide at a range of E:T ratios. In addition, both TCR-transduced T cell populations effectively lysed JCV-infected astrocytes.

ConclusionsWe identified JCV VP1 epitopes that are immunogenic in the context of HLA-A2 MHC-I, including epitopes that have not been previously described. The VP1(100-108) epitope was used to isolate HLA-A2-restricted TCRs. When cloned into primary human CD8+ T cells, these TCRs recognized VP1 (100-108)-presenting targets, and the transduced T cells conferred cytotoxic activity and eliminated K562 and astrocyte cells displaying the VP1(100-108) peptide and not sham peptide, as well as JCV-infected astrocytes. Taken together, these data suggest that JCV VP1-specific TCRs could be appealing therapeutics for HLA-A2+ individuals with PML in whom intrinsic T cell immunity cannot be rescued.
]]></description>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Martinov, T.</dc:creator>
<dc:creator>Thelen, A.</dc:creator>
<dc:creator>Sunahara, M.</dc:creator>
<dc:creator>Mureli, S.</dc:creator>
<dc:creator>Vazquez, A.</dc:creator>
<dc:creator>Gerdts, J.</dc:creator>
<dc:creator>Dandekar, R.</dc:creator>
<dc:creator>Cortese, I.</dc:creator>
<dc:creator>Fouassier, C.</dc:creator>
<dc:creator>Schanzer, E.</dc:creator>
<dc:creator>Urnov, F. D.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Shy, B. R.</dc:creator>
<dc:creator>Greenberg, P. D.</dc:creator>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:date>2024-11-04</dc:date>
<dc:identifier>doi:10.1101/2024.11.04.621904</dc:identifier>
<dc:title><![CDATA[Antigen-Specific T Cell Receptor Discovery for Treating Progressive Multifocal Leukoencephalopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.04.621665v1?rss=1">
<title>
<![CDATA[
Corticonigral projections recruit substantia nigra pars lateralis dopaminergic neurons for auditory threat memories 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.04.621665v1?rss=1</link>
<description><![CDATA[
Dopaminergic neurons (DANs) in the lateral substantia nigra project to the tail of striatum (TS), which is involved in threat conditioning. Auditory cortex also contributes to threatening behaviors, but whether it directly interacts with midbrain DANs and how these interactions might influence threat conditioning remain unclear. Here, functional mapping revealed robust excitatory input from auditory and temporal association cortexes to substantia nigra pars lateralis (SNL) DANs, but not to pars compacta (SNc) DANs. SNL DANs exhibited unique firing patterns, with irregular pacemaking and higher maximal firing, reflecting different channel complements than SNc DANs. Behaviorally, inhibiting cortex to SNL projections impaired memory retrieval during auditory threat conditioning. Thus, we demonstrate robust corticonigral projections to SNL DANs, contrasting with previous observations of sparse cortical input to substantia nigra DANs. These findings distinguish SNL DANs from other nigral populations, highlighting their role in threatening behaviors and expanding knowledge of cortex to midbrain interactions.
]]></description>
<dc:creator>Sansalone, L.</dc:creator>
<dc:creator>Evans, R. C.</dc:creator>
<dc:creator>Twedell, E.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Khaliq, Z. M.</dc:creator>
<dc:date>2024-11-05</dc:date>
<dc:identifier>doi:10.1101/2024.11.04.621665</dc:identifier>
<dc:title><![CDATA[Corticonigral projections recruit substantia nigra pars lateralis dopaminergic neurons for auditory threat memories]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.05.622029v1?rss=1">
<title>
<![CDATA[
Pseudomonas aeruginosa essential gene perturbations that confer vulnerability to the mammalian host environment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.05.622029v1?rss=1</link>
<description><![CDATA[
Multidrug-resistant Pseudomonas aeruginosa causes highly morbid infections that are challenging to treat. While antibiotics reduce bacterial populations during infection, the host environment also plays a key role in inhibiting and eliminating pathogens. Identifying genetic targets that create vulnerabilities to the host environment may uncover strategies to synergize with nutrient limitation or inherent immune processes to clear bacterial infections. Here, we screened a partial knockdown library targeting P. aeruginosa essential and conditionally essential genes in a murine pneumonia model to identify genes with increased vulnerability in the host environment. We found that partial CRISPR interference (CRISPRi) knockdown of 178 genes showed significant fitness defects in mice relative to axenic culture. We validated two important outliers: ispD, encoding a key enzyme in isoprenoid precursor biosynthesis, and pgsA, encoding an enzyme involved in phospholipid synthesis that is strongly upregulated in human infections. Partial knockdown of both genes showed decreased virulence in a mouse survival assay but had little impact on in vitro growth. The use of CRISPRi screening to uncover genetic vulnerabilities represents a promising strategy to prioritize antibacterial targets that interact with the host environment.
]]></description>
<dc:creator>Prasad, N. K.</dc:creator>
<dc:creator>Ward, R. D.</dc:creator>
<dc:creator>Yu, M. A.</dc:creator>
<dc:creator>Kwon, M. S.</dc:creator>
<dc:creator>Banta, A. B.</dc:creator>
<dc:creator>Rosenberg, O. S.</dc:creator>
<dc:creator>Peters, J. M.</dc:creator>
<dc:date>2024-11-05</dc:date>
<dc:identifier>doi:10.1101/2024.11.05.622029</dc:identifier>
<dc:title><![CDATA[Pseudomonas aeruginosa essential gene perturbations that confer vulnerability to the mammalian host environment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.06.621353v1?rss=1">
<title>
<![CDATA[
De novo structural variants in autism spectrum disorder disrupt distal regulatory interactions of neuronal genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.06.621353v1?rss=1</link>
<description><![CDATA[
Three-dimensional genome organization plays a critical role in gene regulation, and disruptions can lead to developmental disorders by altering the contact between genes and their distal regulatory elements. Structural variants (SVs) can disturb local genome organization, such as the merging of topologically associating domains upon boundary deletion. Testing large numbers of SVs experimentally for their effects on chromatin structure and gene expression is time and cost prohibitive. To address this, we propose a computational approach to predict SV impacts on genome folding, which can help prioritize causal hypotheses for functional testing. We developed a weighted scoring method that measures chromatin contact changes specifically affecting regions of interest, such as regulatory elements or promoters, and implemented it in the SuPreMo-Akita software (Gjoni and Pollard 2024). With this tool, we ranked hundreds of de novo SVs (dnSVs) from autism spectrum disorder (ASD) individuals and their unaffected siblings based on predicted disruptions to nearby neuronal regulatory interactions. This revealed that putative cisregulatory element interactions (CREints) are more disrupted by dnSVs from ASD probands versus unaffected siblings. We prioritized candidate variants that disrupt ASD CREints and validated our top-ranked locus using isogenic excitatory neurons with and without the dnSV, confirming accurate predictions of disrupted chromatin contacts. This study establishes disrupted genome folding as a potential genetic mechanism in ASD and provides a general strategy for prioritizing variants predicted to disrupt regulatory interactions across tissues.
]]></description>
<dc:creator>Gjoni, K.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Everitt, A.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2024-11-07</dc:date>
<dc:identifier>doi:10.1101/2024.11.06.621353</dc:identifier>
<dc:title><![CDATA[De novo structural variants in autism spectrum disorder disrupt distal regulatory interactions of neuronal genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.06.622302v1?rss=1">
<title>
<![CDATA[
Long-term effects of psilocybin on dynamic and effectivity connectivity of fronto-striatal-thalamic circuits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.06.622302v1?rss=1</link>
<description><![CDATA[
Psilocybin has been shown to induce fast and sustained improvements in mental well-being across various populations, yet its long-term mechanisms of action are not fully understood. Initial evidence suggests that longitudinal functional and structural brain changes implicate fronto-striatal-thalamic (FST) circuitry, a broad system involved in goal-directed behavior and motivational states. Here, we apply empirical methods and computational modeling to resting-state fMRI data from a within-subject longitudinal psilocybin trial in psychedelic-naive healthy volunteers. We first show increases in FST dynamic activity four weeks after a full dose of psilocybin. We then proceed to mechanistically account for these increased dynamics, by showing that reduced structural constraints underlie increased FST dynamic activity post psilocybin. Further, we show that these reduced structural constraints come along with increased bottom-up and reduced top-down modulation of FST circuits. While cortical reductions in top-down modulation are linked to regional 5-HT2A receptor availability, increased information outflow via subcortical and limbic regions relate to local D2 receptor availability. Together, these findings show that increased FST flexibility weeks after psilocybin administration is linked to serotonergic-mediated decreases in top-down information flow and dopaminergic-mediated increases in bottom-up information flow. This long-term functional re-organization of FST circuits may represent a common mechanism underling the potential clinical efficacy of psilocybin across various neuropsychiatric disorders including substance abuse, major depression, and anorexia.

Significance StatementFronto-striatal-thalamic systems, which underlie motivation and reward, go through profound functional and structural changes following psilocybin administration. We leveraged longitudinal fMRI data from a within-subject psilocybin trial in psychedelic-naive healthy participants to show that psilocybin increases fronto-striatal-thalamic dynamic activity as well as flexibility four weeks after dosing. Computational modeling revealed that this increased flexibility is mechanistically caused by reduced structural constraints on functional dynamics. Further long-term changes included increased bottom-up and reduced top-down information flow mediated by the serotonergic and dopaminergic systems. This long-term functional re-organization of fronto-striatal-thalamic circuits may reflect a common mechanism underlying clinical symptoms improvements across diagnostic groups, such as increased openness, improved well-being, and reductions in anhedonia, apathy, and substance craving.
]]></description>
<dc:creator>Pasquini, L.</dc:creator>
<dc:creator>Vohryzek, J.</dc:creator>
<dc:creator>Escrichs, A.</dc:creator>
<dc:creator>Sanz Perl, Y.</dc:creator>
<dc:creator>Alvarez, A.</dc:creator>
<dc:creator>Idesis, S.</dc:creator>
<dc:creator>Girn, M.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Mitchell, J. M.</dc:creator>
<dc:creator>Gazzaley, A.</dc:creator>
<dc:creator>Kringelbach, M.</dc:creator>
<dc:creator>Nutt, D. J.</dc:creator>
<dc:creator>Lyons, T.</dc:creator>
<dc:creator>Carhart-Harris, R. L.</dc:creator>
<dc:creator>Deco, G.</dc:creator>
<dc:date>2024-11-07</dc:date>
<dc:identifier>doi:10.1101/2024.11.06.622302</dc:identifier>
<dc:title><![CDATA[Long-term effects of psilocybin on dynamic and effectivity connectivity of fronto-striatal-thalamic circuits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.06.622380v1?rss=1">
<title>
<![CDATA[
Acetic acid induces osmotic imbalances in drug-resistant bacteria synergistically enhancing cobalt-doped carbon quantum dots bactericidal efficiency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.06.622380v1?rss=1</link>
<description><![CDATA[
When pathogenic bacteria colonise a wound, they can create an alkaline ecological niche which selects for their survival by creating an inflammatory environment which restricts healthy wound healing to proceed. To aid healing, wound acidification has been exploited to disrupt this process and stimulate fibroblast growth, increase wound oxygen concentrations, minimise proteolytic activity and re-stimulate the host immune system. Within this study, we have developed unique cobalt doped carbon quantum dot nanoparticles which work together with mild acetic acid creating a potent synergistic antimicrobial therapy. The acidic environment alters the osmotic balance of microorganisms forcing them to swell and speed up the internalisation of the ultra-small particles. The particles hyperpolarise the bacterial membranes and generate damaging peroxidase species resulting in cellular lysis. In mice, cobalt doped carbon quantum dots remove MRSA infection while allowing wounds to heal at equivalent rates to uninfected wounds. This work demonstrates how synergistic antimicrobial treatment strategies can be successfully used to combat antimicrobial resistant infections.
]]></description>
<dc:creator>Truskewycz, A.</dc:creator>
<dc:creator>Choi, B.</dc:creator>
<dc:creator>Pedersen, L.</dc:creator>
<dc:creator>Han, J.</dc:creator>
<dc:creator>Halberg, N.</dc:creator>
<dc:date>2024-11-07</dc:date>
<dc:identifier>doi:10.1101/2024.11.06.622380</dc:identifier>
<dc:title><![CDATA[Acetic acid induces osmotic imbalances in drug-resistant bacteria synergistically enhancing cobalt-doped carbon quantum dots bactericidal efficiency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.05.622086v1?rss=1">
<title>
<![CDATA[
Integrated, high-dimensional analysis of CD4 T cell epitope specificities and phenotypes reveals unexpected diversity in the response to Mycobacterium tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.05.622086v1?rss=1</link>
<description><![CDATA[
Immunity to Mycobacterium tuberculosis (Mtb), like many pathogens, is encoded jointly by the antigen specificities and functions of responding CD4 T cells. However, these features span a large two-dimensional possibility space - defined on one axis by the Mtb proteome, and on the other by the T cell transcriptome - that exceeds the dimensionality of existing technologies. Here we present an approach ("CRESTA") that combines highly-multiplexed DNA-barcoded epitope probes, single cell sequencing, and clonal analysis of T Cell Receptors (TCRs) to robustly detect rare antigen-specific CD4 T cells across hundreds of epitopes simultaneously and reveal their transcriptome-wide phenotypes. By comprehensively assaying known epitopes in Mtb-infected participants, we reveal polyclonal and multi-epitope responses across a spectrum of differentiation states, uncover previously-unobserved phenotypic diversity within and between epitopes, and increase the total number of known Mtb epitope-mapped TCR:{beta}s by [~]8-fold. We expect CRESTA to enable high-dimensional analyses of CD4 T cell responses in various settings, including infection, cancer, autoimmunity and allergy.
]]></description>
<dc:creator>Mead, H. L.</dc:creator>
<dc:creator>Kirschman, J. H.</dc:creator>
<dc:creator>Harms, C. E.</dc:creator>
<dc:creator>Kelley, E. J.</dc:creator>
<dc:creator>Soria-Bustos, J.</dc:creator>
<dc:creator>Nelson, G. A.</dc:creator>
<dc:creator>Ogongo, P.</dc:creator>
<dc:creator>Ouma, G.</dc:creator>
<dc:creator>Ouma, S. G.</dc:creator>
<dc:creator>Ernst, J. D.</dc:creator>
<dc:creator>Altin, J. A.</dc:creator>
<dc:date>2024-11-08</dc:date>
<dc:identifier>doi:10.1101/2024.11.05.622086</dc:identifier>
<dc:title><![CDATA[Integrated, high-dimensional analysis of CD4 T cell epitope specificities and phenotypes reveals unexpected diversity in the response to Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.06.622278v1?rss=1">
<title>
<![CDATA[
Human xenobiotic metabolism proteins have full-length and split homologs in the gut microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.06.622278v1?rss=1</link>
<description><![CDATA[
Xenobiotics, including pharmaceutical drugs, can be metabolized by both host and microbiota, in some cases by homologous enzymes. We conducted a systematic search for all human proteins with gut microbial homologs. Because gene fusion and fission can obscure homology detection, we built a pipeline to identify not only full-length homologs, but also cases where microbial homologs were split across multiple adjacent genes in the same neighborhood or operon ("split homologs"). We found that human proteins with full-length gut microbial homologs disproportionately participate in xenobiotic metabolism. While this included many different enzyme classes, short-chain and aldo-keto reductases were the most frequently detected, especially in prevalent gut microbes, while cytochrome P450 homologs were largely restricted to lower-prevalence facultative anaerobes. In contrast, human proteins with split homologs tended to play roles in central metabolism, especially of nucleobase-containing compounds. We identify twelve specific drugs that gut microbial split homologs may metabolize; two of these, 6-mercaptopurine by xanthine dehydrogenase (XDH) and 5-fluorouracil by dihydropyrimidine dehydrogenase (DPYD), have been recently confirmed in mouse models. This work provides a comprehensive map of homology between the human and gut microbial proteomes, indicates which human xenobiotic enzyme classes are most likely to be shared by gut microorganisms, and finally demonstrates that split homology may be an underappreciated explanation for microbial contributions to drug metabolism.

Article SummaryWe develop a pipeline to systematically find human proteins with gut microbial homologs, including those split across multiple microbial genes (e.g., operons). This reveals thousands of proteins with full-length gut homologs, especially reductases and hydrolases that metabolize xenobiotics. Nearly two dozen split homologs are also observed for central metabolic enzymes, many of which can transform substrate analogs; in two cases, previous studies verify that microbial split homologs enable the expected drug to be metabolized in vivo. These results, which we provide as a resource, map out homology and shed light on parallel drug metabolism between host and microbiome.
]]></description>
<dc:creator>Rendina, M.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:date>2024-11-08</dc:date>
<dc:identifier>doi:10.1101/2024.11.06.622278</dc:identifier>
<dc:title><![CDATA[Human xenobiotic metabolism proteins have full-length and split homologs in the gut microbiome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.06.622344v1?rss=1">
<title>
<![CDATA[
Evolutionarily guided transcription factor design programs novel T cell states 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.06.622344v1?rss=1</link>
<description><![CDATA[
Protein-coding genes in the human genome evolved via modular rearrangement of domains from ancestral genes1. Here, we develop a scalable, evolutionarily guided method to assemble novel protein-coding genes from constituent domains within a protein family, termed DESynR (Domain Engineered via Synthesis and Recombination) genes. Using primary human chimeric antigen receptor T cells as a model system, we find that the expression of DESynR Activator Protein-1 (AP-1) transcription factors (TFs) significantly outperforms the overexpression of natural AP-1 TFs in multiple functional assays in vitro and in vivo. Top DESynR AP-1 TFs exhibit non-intuitive architectures of constituent domains, including from TFs that are not canonically expressed in T cells. DESynR AP-1 TFs induce broad transcriptional and epigenetic reprogramming of T cells and, in some cases, lead to the development of non-natural T cell states, engaging gene expression modules from disparate human cell types. Taken together, we demonstrate that novel configurations of existing protein domains may uncover non-evolved genes that program cell states with therapeutically relevant functions.
]]></description>
<dc:creator>Takacsi-Nagy, O.</dc:creator>
<dc:creator>Hartman, A.</dc:creator>
<dc:creator>Chen, A. Y.</dc:creator>
<dc:creator>Yin, Y.</dc:creator>
<dc:creator>Reeder, G. C.</dc:creator>
<dc:creator>Kernick, C.</dc:creator>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>McClellan, A. K.</dc:creator>
<dc:creator>Raposo, C. J.</dc:creator>
<dc:creator>Theberath, N. E.</dc:creator>
<dc:creator>Yan, P. K.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Roth, T. L.</dc:creator>
<dc:creator>Satpathy, A. T.</dc:creator>
<dc:date>2024-11-08</dc:date>
<dc:identifier>doi:10.1101/2024.11.06.622344</dc:identifier>
<dc:title><![CDATA[Evolutionarily guided transcription factor design programs novel T cell states]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.07.622474v1?rss=1">
<title>
<![CDATA[
Genetic Basis, Quantitative Nature, and Functional Relevance of Evolutionarily Conserved DNA Methylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.07.622474v1?rss=1</link>
<description><![CDATA[
DNA methylation (DNAm) is a key epigenetic mark that modulates regulatory elements and gene expression, playing a crucial role in mammalian development and physiological function. Despite extensive characterization of DNAm profiles across species, little is known about its evolutionary conservation. Here, we conducted a comparative epigenome-wide analysis of great apes to identify and characterize sequence- and methylation-conserved CpGs (MCCs). Using 202 DNAm arrays, alongside 6 matched genotype and 13 matched transcriptomic datasets, we identified 11,500 MCCs for which methylation was evolutionarily related to sequences of CpGs and methylation quantitative trait loci. MCCs were the most stable across healthy human tissues and exhibited weaker genetic associations than other CpGs. Moreover, MCCs showed minimal associations with demographic, environmental factors, and noncancer diseases, yet demonstrated stronger associations with certain cancers than other CpGs, particularly gastrointestinal cancers. Functional enrichment analysis revealed that genes associated with MCC methylation in cancer were enriched for cancer driver genes and canonical cancer pathways, highlighting a significant regulatory role for MCCs in tumorigenesis. Collectively, our findings reveal the extent of DNAm conservation in great ape evolution, its association with genetic conservation, and its relevance to human diseases. These integrative analyses offer evolutionary insights into epigenetic variation and its functional implications in human populations.
]]></description>
<dc:creator>Dong, Z.</dc:creator>
<dc:creator>Schaffner, S.</dc:creator>
<dc:creator>Fu, M.</dc:creator>
<dc:creator>Whitehead, J.</dc:creator>
<dc:creator>MacIsaac, J. L.</dc:creator>
<dc:creator>Rehkopf, D. H.</dc:creator>
<dc:creator>Boyce, W. T.</dc:creator>
<dc:creator>Rosero-Bixby, L.</dc:creator>
<dc:creator>Quintana-Murci, L.</dc:creator>
<dc:creator>Patin, E.</dc:creator>
<dc:creator>Miller, G. E.</dc:creator>
<dc:creator>Korthauer, K.</dc:creator>
<dc:creator>Kobor, M. S.</dc:creator>
<dc:date>2024-11-09</dc:date>
<dc:identifier>doi:10.1101/2024.11.07.622474</dc:identifier>
<dc:title><![CDATA[Genetic Basis, Quantitative Nature, and Functional Relevance of Evolutionarily Conserved DNA Methylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.07.622542v1?rss=1">
<title>
<![CDATA[
Disease-Linked Regulatory DNA Variants and Homeostatic Transcription Factors in Epidermis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.07.622542v1?rss=1</link>
<description><![CDATA[
Identifying noncoding single nucleotide variants (SNVs) in regulatory DNA linked to polygenic disease risk, the transcription factors (TFs) they bind, and the target genes they dysregulate is a goal in polygenic disease research. Massively parallel reporter gene analysis (MPRA) of 3,451 SNVs linked to risk for polygenic skin diseases characterized by disrupted epidermal homeostasis identified 355 differentially active SNVs (daSNVs). daSNV target gene analysis, combined with daSNV editing, underscored dysregulated epidermal differentiation as a pathomechanism shared across common polygenic skin diseases. CRISPR knockout screens of 1772 human TFs revealed 108 TFs essential for epidermal progenitor differentiation, uncovering novel roles for ZNF217, CXXC1, FOXJ2, IRX2 and NRF1. Population sampling CUT&RUN of 27 homeostatic TFs identified allele-specific DNA binding (ASB) differences at daSNVs enriched near epidermal homeostasis and monogenic skin disease genes, with notable representation of SP/KLF and AP-1/2 TFs. This resource implicates dysregulated differentiation in risk for diverse polygenic skin diseases.
]]></description>
<dc:creator>Porter, D. F.</dc:creator>
<dc:creator>Meyers, R. M.</dc:creator>
<dc:creator>Miao, W.</dc:creator>
<dc:creator>Reynolds, D.</dc:creator>
<dc:creator>Hong, A. W.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Mondal, S.</dc:creator>
<dc:creator>Siprashvili, Z.</dc:creator>
<dc:creator>Srinivasan, S.</dc:creator>
<dc:creator>DUCOLI, L.</dc:creator>
<dc:creator>Meyers, J.</dc:creator>
<dc:creator>Nguyen, D.</dc:creator>
<dc:creator>Ko, L.</dc:creator>
<dc:creator>Kellman, L.</dc:creator>
<dc:creator>Elfaki, I.</dc:creator>
<dc:creator>Guo, M.</dc:creator>
<dc:creator>Winge, M.</dc:creator>
<dc:creator>Lopez-Pajares, V.</dc:creator>
<dc:creator>Tao, S.</dc:creator>
<dc:creator>Khavari, P.</dc:creator>
<dc:date>2024-11-11</dc:date>
<dc:identifier>doi:10.1101/2024.11.07.622542</dc:identifier>
<dc:title><![CDATA[Disease-Linked Regulatory DNA Variants and Homeostatic Transcription Factors in Epidermis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.11.623014v1?rss=1">
<title>
<![CDATA[
Single-cell spatial transcriptomics of fixed, paraffin-embedded biopsies reveals colitis-associated cell networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.11.623014v1?rss=1</link>
<description><![CDATA[
Background & AimsImaging-based, single-cell spatial transcriptomics (iSCST) using formalin-fixed, paraffin-embedded (FFPE) tissue could transform translational research by retaining all tissue cell subsets and spatial locations while enabling the analysis of archived specimens. We aimed to develop a robust framework for applying iSCST to archived clinical FFPE mucosal biopsies from patients with inflammatory bowel disease (IBD).

MethodsWe performed a comprehensive benchmarking comparison of three iSCST platforms capable of analyzing FFPE specimens. We analyzed FFPE mucosal biopsies (n=57) up to 5 years old from non-IBD controls (HC; n=9) and patients with ulcerative colitis (UC;n=11). After platform-specific cell segmentation, we applied a uniform data processing pipeline to all datasets, including transcript detection, cell annotation, differential gene expression, and neighborhood enrichment. Transcriptomic signatures identified with iSCST were validated using external, publicly available bulk transcriptomic datasets.

ResultsA custom 290-plex Xenium gene panel exhibited the highest sensitivity and specificity for transcript detection, enabling precise identification and quantification of diverse cell subsets and differentially expressed genes across cell types and disease states. We mapped transcriptionally distinct fibroblast subsets to discrete spatial locations and identified inflammation-associated fibroblasts (IAFs) and monocytes as a colitis-associated cellular neighborhood. We also identified signatures associated with Vedolizumab (VDZ) responsiveness. VDZ non-responders were characterized by an IAF-monocyte transcriptional signature, while responders exhibited enrichment of epithelial gene sets.

ConclusionsOur optimized iSCST framework for archived FFPE biopsies provides unique advantages for assessing the role of colitis-associated cellular networks in routinely collected clinical samples. FFPE-based biomarkers could integrate with existing clinical workflows and potentially aid in risk-stratifying patients.
]]></description>
<dc:creator>Mennillo, E.</dc:creator>
<dc:creator>Lotstein, M. L.</dc:creator>
<dc:creator>Lee, G.</dc:creator>
<dc:creator>Johri, V.</dc:creator>
<dc:creator>Ekstrand, C.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Hou, J.</dc:creator>
<dc:creator>Leet, D. E.</dc:creator>
<dc:creator>He, J. Y.</dc:creator>
<dc:creator>Mahadevan, U.</dc:creator>
<dc:creator>Eckalbar, W. L.</dc:creator>
<dc:creator>Oh, D. Y.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Kattah, M. G.</dc:creator>
<dc:creator>Combes, A.</dc:creator>
<dc:date>2024-11-11</dc:date>
<dc:identifier>doi:10.1101/2024.11.11.623014</dc:identifier>
<dc:title><![CDATA[Single-cell spatial transcriptomics of fixed, paraffin-embedded biopsies reveals colitis-associated cell networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.11.623012v1?rss=1">
<title>
<![CDATA[
The Impact of Charge Regulation and Ionic Intranuclear Environment on the Nucleosome Core Particle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.11.623012v1?rss=1</link>
<description><![CDATA[
We theoretically investigate how the intranuclear environment influences the charge of a nucleosome core particle (NCP) - the fundamental unit of chromatin consisting of DNA wrapped around a core of histone proteins. The molecular-based theory explicitly considers the size, shape, conformations, charges, and chemical states of all molecular species - thereby linking the structural state with the chemical/charged state of the system. We investigate how variations in monovalent and divalent salt concentrations, as well as pH, affect the charge distribution across different regions of an NCP and quantify the impact of charge regulation. The effective charge of an NCP emerges from a delicate and complex balance involving the chemical dissociation equilibrium of the amino acids and the DNA-phosphates, the electrostatic interaction between them, and the translational entropy of the mobile solution ions, i.e., counter ion release and ion condensation. From our results, we note the significant effect of divalent magnesium ions on the charge and electrostatic energy as well as the counterion cloud that surrounds an NCP, as a function of magnesium concentration, charge neutralization, and even charge inversion is predicted - in line with experimental observation of NCPs. The strong Mg-dependence of the nucleosome charge state arises from ion bridges between two DNA-phosphates and one Mg + ion. We demonstrate that to describe and predict the charged state of an NCP properly, it is essential to consider molecular details, such as DNA-phosphate ion condensation and the acid-base equilibrium of the amino acids that comprise the core histone proteins.
]]></description>
<dc:creator>Nap, R. J.</dc:creator>
<dc:creator>Gonzalez, P. C.</dc:creator>
<dc:creator>Coraor, A. E.</dc:creator>
<dc:creator>Virk, R. K. A.</dc:creator>
<dc:creator>de Pablo, J. J.</dc:creator>
<dc:creator>Backman, V.</dc:creator>
<dc:creator>Szleifer, I.</dc:creator>
<dc:date>2024-11-12</dc:date>
<dc:identifier>doi:10.1101/2024.11.11.623012</dc:identifier>
<dc:title><![CDATA[The Impact of Charge Regulation and Ionic Intranuclear Environment on the Nucleosome Core Particle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.12.623285v1?rss=1">
<title>
<![CDATA[
Loss of TMEM55B modulates lipid metabolism through dysregulated lipophagy and mitochondrial function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.12.623285v1?rss=1</link>
<description><![CDATA[
Lipophagy is a form of selective autophagy that targets the lipid droplets for lysosomal decay and has been implicated in the onset and progression of metabolic dysfunction-associated steatotic liver disease (MASLD). Factors that augment lipophagy have been identified as targets for MASLD therapeutic development. TMEM55B is a key regulator of lysosomal positioning which is critical for the lysosome fusion with the autophagosome, but less studied. Here, we demonstrate that inhibition of TMEM55B in murine models accelerates MASLD onset and progression. In cellular models, TMEM55B deficiency enhances lipophagy, leading to increased fatty acid release from lysosomes to mitochondria but simultaneously impairs mitophagy, causing an accumulation of dysfunctional mitochondria. This imbalance leads to increased lipid accumulation and oxidative stress, worsening MASLD. These findings underscore the importance of lysosomal positioning in lipid metabolism and suggest that augmenting lipophagy may exacerbate disease in the context of mitochondrial dysfunction.
]]></description>
<dc:creator>Qin, Y.</dc:creator>
<dc:creator>Teker, S.</dc:creator>
<dc:creator>La Cunza, N.</dc:creator>
<dc:creator>Theusch, E.</dc:creator>
<dc:creator>Yang, N.</dc:creator>
<dc:creator>Venkatesan, L.</dc:creator>
<dc:creator>Su, J.</dc:creator>
<dc:creator>Krauss, R. M.</dc:creator>
<dc:creator>Lakkaraju, A.</dc:creator>
<dc:creator>Mattis, A. N.</dc:creator>
<dc:creator>Medina, M. W.</dc:creator>
<dc:date>2024-11-13</dc:date>
<dc:identifier>doi:10.1101/2024.11.12.623285</dc:identifier>
<dc:title><![CDATA[Loss of TMEM55B modulates lipid metabolism through dysregulated lipophagy and mitochondrial function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.11.623079v1?rss=1">
<title>
<![CDATA[
Sulfur-species in Zinc-specific Condylar Zones of a Rat Temporomandibular Joint 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.11.623079v1?rss=1</link>
<description><![CDATA[
In this study, we performed synchrotron-based micro-X-ray fluorescence (-XRF) imaging of elements Zn and S, and X-ray absorption near edge spectroscopy (XANES) coupled with -XRF for identification of Zn and S species in the condylar zones of a rat temporomandibular joint (TMJ). Histologic localization of Zn and hypoxia-inducible factor-1 (HIF-1) were mapped using an optical microscope. These data were visually correlated with -XRF and XANES data to provide insights into plausible biological S-species in Z-enriched condylar zones of a rat TMJ. Furthermore, -XRF coupled with micro-X-ray diffraction (-XRD) was used to underline Z-incorporated biological apatite in the subchondral bone and bone of the rat TMJ.

Results illustrated the potential dependence between biometal Zn and nonmetal S and their collective governance of cell and tissue functions in a zone-specific manner. Elemental Zn with organic and inorganic S-species at the cartilage-bone interface and transformation of plausible Zn-enriched mineralization kinetics of biological apatite from subchondral bone to condylar bone were ascertained using -XRF-XANES and -XRD. The coupled -XRF-XANES complementing with -XRD and immunohistology provided an informative view of S and Zn and their association with zone-specific biological pathways in situ. Understanding the spatial distributions of the main S-species with redox-inert Zn in regions of cartilage, bone, and the interface is essential for further unlocking questions surrounding formation and resorption-related biomineralization pathways as related to osteoarthritis or genetically inherited diseases. Using these complementary techniques with microspectroscopic spatial information provided insights into the associations between biometal Zn and nonmetal S and a window into detecting the plausible early-stage diagnostic biomarkers for humans with TMJ osteoarthritis.
]]></description>
<dc:creator>Lee, B. H.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Ho, T. J.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Tamura, N.</dc:creator>
<dc:creator>Webb, S.</dc:creator>
<dc:creator>Bone, S.</dc:creator>
<dc:creator>Ho, S. P.</dc:creator>
<dc:date>2024-11-14</dc:date>
<dc:identifier>doi:10.1101/2024.11.11.623079</dc:identifier>
<dc:title><![CDATA[Sulfur-species in Zinc-specific Condylar Zones of a Rat Temporomandibular Joint]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.14.623592v1?rss=1">
<title>
<![CDATA[
Interspecies Organoids Reveal Human-Specific Molecular Features of Dopaminergic Neuron Development and Vulnerability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.14.623592v1?rss=1</link>
<description><![CDATA[
The disproportionate expansion of telencephalic structures during human evolution involved tradeoffs that imposed greater connectivity and metabolic demands on midbrain dopaminergic neurons. Despite the central role of dopaminergic neurons in human-enriched disorders, molecular specializations associated with human-specific features and vulnerabilities of the dopaminergic system remain unexplored. Here, we establish a phylogeny-in-a-dish approach to examine gene regulatory evolution by differentiating pools of human, chimpanzee, orangutan, and macaque pluripotent stem cells into ventral midbrain organoids capable of forming long-range projections, spontaneous activity, and dopamine release. We identify human-specific gene expression changes related to axonal transport of mitochondria and reactive oxygen species buffering and candidate cis- and trans-regulatory mechanisms underlying gene expression divergence. Our findings are consistent with a model of evolved neuroprotection in response to tradeoffs related to brain expansion and could contribute to the discovery of therapeutic targets and strategies for treating disorders involving the dopaminergic system.
]]></description>
<dc:creator>Nolbrant, S.</dc:creator>
<dc:creator>Wallace, J. L.</dc:creator>
<dc:creator>Ding, J.</dc:creator>
<dc:creator>Zhu, T.</dc:creator>
<dc:creator>Sevetson, J. L.</dc:creator>
<dc:creator>Kajtez, J.</dc:creator>
<dc:creator>Baldacci, I. A.</dc:creator>
<dc:creator>Corrigan, E. K.</dc:creator>
<dc:creator>Hoglin, K.</dc:creator>
<dc:creator>McMullen, R.</dc:creator>
<dc:creator>Schmitz, M. T.</dc:creator>
<dc:creator>Breevoort, A.</dc:creator>
<dc:creator>Swope, D.</dc:creator>
<dc:creator>Wu, F.</dc:creator>
<dc:creator>Pavlovic, B. J.</dc:creator>
<dc:creator>Salama, S. R.</dc:creator>
<dc:creator>Kirkeby, A.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Schaefer, N. K.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:date>2024-11-15</dc:date>
<dc:identifier>doi:10.1101/2024.11.14.623592</dc:identifier>
<dc:title><![CDATA[Interspecies Organoids Reveal Human-Specific Molecular Features of Dopaminergic Neuron Development and Vulnerability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.14.623693v1?rss=1">
<title>
<![CDATA[
Reduction of DNA Topoisomerase Top2 reprograms the epigenetic landscape and extends health and life span across species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.14.623693v1?rss=1</link>
<description><![CDATA[
DNA topoisomerases are essential molecular machines that manage DNA topology in the cell and play important roles in DNA replication and transcription. We find that knocking down the enzyme Topoisomerase Top2 or its mammalian homolog Top2b increases the life span of S. cerevisiae, C. elegans, and mice. Top2b reduction also extends the health span of mice and alleviates the pathologies of aging in multiple tissues. At the cellular/molecular level, Top2b reduction attenuates the major hallmarks of aging, such as cellular senescence, de-regulated nutrient sensing, epigenetic alteration, and lysosomal biogenesis. We observed that Top2b reduction significantly changes the epigenetic landscape in various mouse tissues toward those of the young animals, and differentially down-regulates genes with active promoter and high expression. Our observations suggest that Top2 reduction confers longevity effect across species via a conserved mechanism, and may be used as a novel therapeutic strategy for countering aging.
]]></description>
<dc:creator>Zhu, M.</dc:creator>
<dc:creator>Ma, M.</dc:creator>
<dc:creator>Luo, L.</dc:creator>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Pan, Y.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Xiao, Y.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Xian, B.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:date>2024-11-15</dc:date>
<dc:identifier>doi:10.1101/2024.11.14.623693</dc:identifier>
<dc:title><![CDATA[Reduction of DNA Topoisomerase Top2 reprograms the epigenetic landscape and extends health and life span across species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.18.624137v1?rss=1">
<title>
<![CDATA[
Inter- and Intrahemispheric Sources of Vestibular Signals to V1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.18.624137v1?rss=1</link>
<description><![CDATA[
Head movements are sensed by the vestibular organs. Unlike classical senses, signals from vestibular organs are not conveyed to a dedicated cortical area but are broadcast throughout the cortex. Surprisingly, the routes taken by vestibular signals to reach the cortex are still largely uncharted. Here we show that the primary visual cortex (V1) receives real-time head movement signals -- direction, velocity, and acceleration -- from the ipsilateral pulvinar and contralateral visual cortex. The ipsilateral pulvinar provides the main head movement signal, with a bias toward contraversive movements (e.g. clockwise movements in left V1). Conversely, the contralateral visual cortex provides head movement signals during ipsiversive movements. Crucially, head movement variables encoded in V1 are already encoded in the pulvinar, suggesting that those variables are computed subcortically. Thus, the convergence of inter- and intrahemispheric signals endows V1 with a rich representation of the animals head movements.
]]></description>
<dc:creator>Bouvier, G.</dc:creator>
<dc:creator>Sanzeni, A.</dc:creator>
<dc:creator>Hamada, E.</dc:creator>
<dc:creator>Brunel, N.</dc:creator>
<dc:creator>Scanziani, M.</dc:creator>
<dc:date>2024-11-19</dc:date>
<dc:identifier>doi:10.1101/2024.11.18.624137</dc:identifier>
<dc:title><![CDATA[Inter- and Intrahemispheric Sources of Vestibular Signals to V1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.19.624390v1?rss=1">
<title>
<![CDATA[
Cellular behavior analysis from live-cell imaging of TCR T cell-cancer cell interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.19.624390v1?rss=1</link>
<description><![CDATA[
T cell therapies, such as chimeric antigen receptor (CAR) T cells and T cell receptor (TCR) T cells, are a growing class of anti-cancer treatments. However, expansion to novel indications and beyond last-line treatment requires engineering cells dynamic population behaviors. Here we develop the tools for cellular behavior analysis of T cells from live-cell imaging, a common and inexpensive experimental setup used to evaluate engineered T cells. We first develop a state-of-the-art segmentation and tracking pipeline, Caliban, based on human-in-the-loop deep learning. We then build the Occident pipeline to collect a catalog of phenotypes that characterize cell populations, morphology, movement, and interactions in co-cultures of modified T cells and antigen-presenting tumor cells. We use Caliban and Occident to interrogate how interactions between T cells and cancer cells differ when beneficial knock-outs of RASA2 and CUL5 are introduced into TCR T cells. We apply spatiotemporal models to quantify T cell recruitment and proliferation after interactions with cancer cells. We discover that, compared to a safe harbor knockout control, RASA2 knockout T cells have longer interaction times with cancer cells leading to greater T cell activation and killing efficacy, while CUL5 knockout T cells have increased proliferation rates leading to greater numbers of T cells for hunting. Together, segmentation and tracking from Caliban and phenotype quantification from Occident enable cellular behavior analysis to better engineer T cell therapies for improved cancer treatment.
]]></description>
<dc:creator>Verma, A.</dc:creator>
<dc:creator>Yu, C.</dc:creator>
<dc:creator>Bachl, S.</dc:creator>
<dc:creator>Lopez, I.</dc:creator>
<dc:creator>Schwartz, M. S.</dc:creator>
<dc:creator>Moen, E.</dc:creator>
<dc:creator>Kale, N.</dc:creator>
<dc:creator>Ching, C.</dc:creator>
<dc:creator>Miller, G.</dc:creator>
<dc:creator>Dougherty, T.</dc:creator>
<dc:creator>Pao, E.</dc:creator>
<dc:creator>Graf, W.</dc:creator>
<dc:creator>Ward, C.</dc:creator>
<dc:creator>Jena, S. G.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Carnevale, J.</dc:creator>
<dc:creator>Van Valen, D. A.</dc:creator>
<dc:creator>Engelhardt, B. E.</dc:creator>
<dc:date>2024-11-21</dc:date>
<dc:identifier>doi:10.1101/2024.11.19.624390</dc:identifier>
<dc:title><![CDATA[Cellular behavior analysis from live-cell imaging of TCR T cell-cancer cell interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.19.624397v1?rss=1">
<title>
<![CDATA[
Beta burst characteristics and coupling within the sensorimotor cortical-subthalamic nucleus circuit in Parkinson's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.19.624397v1?rss=1</link>
<description><![CDATA[
BackgroundBursts of exaggerated subthalamic nucleus (STN) beta activity contribute to clinical impairments in Parkinsons disease (PD). Few studies have explored the characteristics and coupling of bursts across the sensorimotor cortical-STN circuit.

ObjectiveWe sought to (1) establish the characteristics of sensorimotor cortical and STN bursts during naturalistic behaviours, and (2) determine the predictability of STN bursts from motor cortical recordings.

MethodsWe analysed 1,478 hours of wirelessly streamed bilateral sensorimotor cortical and STN recordings from 5 PD patients.

ResultsSTN bursts were longer than cortical bursts and had shorter inter-burst intervals. Long bursts (>200ms) in both structures displayed temporal overlap (>30%), with an estimated cortico-STN conduction delay of 8ms. Furthermore, approximately 27% of all STN bursts were preceded by a cortical burst.

ConclusionCortical beta bursts tend to precede STN beta bursts, with short delays. However, subcortical mechanisms are also likely to contribute to STN burst initiation and propagation.
]]></description>
<dc:creator>Yao, P.</dc:creator>
<dc:creator>Abdi-Sargezeh, B.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Tan, H.</dc:creator>
<dc:creator>Hahn, A.</dc:creator>
<dc:creator>Starr, P.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:creator>Oswal, A.</dc:creator>
<dc:date>2024-11-21</dc:date>
<dc:identifier>doi:10.1101/2024.11.19.624397</dc:identifier>
<dc:title><![CDATA[Beta burst characteristics and coupling within the sensorimotor cortical-subthalamic nucleus circuit in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.21.624591v1?rss=1">
<title>
<![CDATA[
Coordinated protein modules define DNA damage responses to carboplatin at single cell resolution in human ovarian carcinoma models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.21.624591v1?rss=1</link>
<description><![CDATA[
Tubo-ovarian high-grade serous carcinoma (HGSC) is the most lethal gynecological malignancy and frequently responds to platinum-based chemotherapy because of common genetic and somatic impairment of DNA damage repair (DDR) pathways. The mechanisms of clinical platinum resistance are diverse and poorly molecularly defined. Consequently, there are no biomarkers or medicines that improve patient outcomes. Herein we use single cell mass cytometry (CyTOF) to systematically evaluate the phosphorylation and abundance of proteins known to participate in the DNA damage response (DDR). Single cell analyses of highly characterized HGSC cell lines that phenocopy human patients show that cells with comparable levels of intranuclear platinum, a proxy for carboplatin uptake, undergo different cell fates. Unsupervised analyses revealed a continuum of DDR responses. Decompositional methods were used to identify eight distinct protein modules of carboplatin resistance and sensitivity at single cell resolution. CyTOF profiling of primary and secondary platinum-resistance patient models shows that a complex DDR sensitivity module is strongly associated with response, suggesting it as a potential tool to clinically characterize complex drug resistance phenotypes.
]]></description>
<dc:creator>Bedia, J. S.</dc:creator>
<dc:creator>Huang, Y.-W.</dc:creator>
<dc:creator>Gonzalez, A. D.</dc:creator>
<dc:creator>Gonzalez, V. D.</dc:creator>
<dc:creator>Funingana, I.-G.</dc:creator>
<dc:creator>Rahil, Z.</dc:creator>
<dc:creator>Mike, A.</dc:creator>
<dc:creator>Lowar, A.</dc:creator>
<dc:creator>Vias, M.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Brenton, J. D.</dc:creator>
<dc:creator>Fantl, W. J.</dc:creator>
<dc:date>2024-11-22</dc:date>
<dc:identifier>doi:10.1101/2024.11.21.624591</dc:identifier>
<dc:title><![CDATA[Coordinated protein modules define DNA damage responses to carboplatin at single cell resolution in human ovarian carcinoma models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.21.624708v1?rss=1">
<title>
<![CDATA[
Dermatopontin-expressing fibroblasts mediate an essential skin macrophage niche 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.21.624708v1?rss=1</link>
<description><![CDATA[
Fibroblasts are present in all tissues and are crucial for maintaining tissue homeostasis. We previously identified fibroblasts marked by Dermatopontin (Dpt) but their role in supporting macrophage homeostasis remains unclear. Here, we generated novel mesenchymal lineage-restricted genetic tools to target Dpt expressing fibroblasts and elucidate their role in supporting skin macrophages. Transcriptional profiling, flow cytometry, and in situ hybridization uncovered two broad populations of F4/80-expressing skin macrophages, denoted by high expression of CD206 and CD64 (CD206hiCD64+), or CD11c. Targeted depletion of Dpt+ fibroblasts resulted in a profound loss of both macrophage populations. Conditional deletion of colony-stimulating factor-1 (Csf1) in Dpt+ fibroblasts revealed that CD206hiCD64+, and not CD11c+, macrophages are acutely dependent on fibroblast-derived Csf1, consistent with their higher expression of the Csf1 receptor. Following Csf1 deletion in Dpt+ fibroblasts, loss of CD206hiCD64+ macrophages were observed across the dermis, dermal white adipose tissue (dWAT), and adventitia, accompanied by a modest upregulation of fibroblast-related and extracellular matrix (ECM) genes and structural changes to the skin. Alterations to the skin network upon loss of fibroblast-derived Csf1 and CD206hiCD64+ macrophages led to a significant delay in wound healing. We also demonstrate the CSF1-CSF1R signaling pathway is functionally relevant in human systemic sclerosis, or scleroderma, as elevated levels of CSF1 produced by fibroblasts and an increased abundance of macrophages both correlate with disease severity. Our findings demonstrate the role of Dpt+ fibroblasts in regulating a Csf1-dependent macrophage niche in skin and orchestrating responses in injury and disease.
]]></description>
<dc:creator>Vollmers, A. C.</dc:creator>
<dc:creator>Wu, S. Z.</dc:creator>
<dc:creator>Altieri, A.</dc:creator>
<dc:creator>McCartney, E. E.</dc:creator>
<dc:creator>Bender, H.</dc:creator>
<dc:creator>Lee, W. P.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Hu, C.</dc:creator>
<dc:creator>Uttarwar, S.</dc:creator>
<dc:creator>Vander Heiden, J. A.</dc:creator>
<dc:creator>Davidson, C.</dc:creator>
<dc:creator>Chung, Y.</dc:creator>
<dc:creator>Ortiz, W.</dc:creator>
<dc:creator>Long, M.</dc:creator>
<dc:creator>Asuncion, R.</dc:creator>
<dc:creator>Yang, Y. A.</dc:creator>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Modrusan, Z.</dc:creator>
<dc:creator>Krishnamurty, A.</dc:creator>
<dc:creator>Fu, W.</dc:creator>
<dc:creator>Müller, S.</dc:creator>
<dc:creator>Buechler, M. B.</dc:creator>
<dc:creator>Turley, S.</dc:creator>
<dc:date>2024-11-22</dc:date>
<dc:identifier>doi:10.1101/2024.11.21.624708</dc:identifier>
<dc:title><![CDATA[Dermatopontin-expressing fibroblasts mediate an essential skin macrophage niche]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.23.625018v1?rss=1">
<title>
<![CDATA[
Blood draw site and blood matrix influence mineral assessment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.23.625018v1?rss=1</link>
<description><![CDATA[
Circulating mineral concentrations are used to assess nutritional status or as markers for clinical conditions, but the quality of these measurements depends on the methods used for blood processing. Since the procedures for measuring mineral levels in blood are not standardized, discrepancies in sampling may influence the analytical results. We previously reported that zinc content in blood samples showed significant variation depending on several pre-analytical variables selected during blood collection and processing. In this study, we extend our analysis to determine how other mineral levels might be affected by different blood draw sites (capillary or venous) or sample matrices (plasma or serum). Sequential capillary and venous blood samples were collected from a diverse cohort of sixty healthy adults and analyzed for multiple minerals using inductively coupled plasma- optical emission spectrometry. Calcium, copper, iron, and magnesium were the only minerals that were detected in all samples and were free from contamination in the blood collection tubes used for the study. When assessing different blood draw sites, the concentrations of calcium, copper, iron, and magnesium were 2-11% higher from capillary compared to the venous plasma. When assessing different blood sample matrices, the concentrations of calcium, copper, and magnesium were 2-5% higher in serum compared to plasma samples, whereas the concentration of iron was 7% higher in plasma compared to serum samples. The differences observed in these four essential minerals from discrepant draw sites and blood matrices demonstrate the importance of controlling key pre-analytic variables when assessing mineral levels in blood.
]]></description>
<dc:creator>Alejandro, M. G.</dc:creator>
<dc:creator>Schultz, K.</dc:creator>
<dc:creator>Killilea, D. W.</dc:creator>
<dc:date>2024-11-24</dc:date>
<dc:identifier>doi:10.1101/2024.11.23.625018</dc:identifier>
<dc:title><![CDATA[Blood draw site and blood matrix influence mineral assessment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.21.624682v1?rss=1">
<title>
<![CDATA[
Replication stress marker phospho-RPA2 predicts response to platinum and PARP inhibitors in homologous recombination-proficient ovarian cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.21.624682v1?rss=1</link>
<description><![CDATA[
BackgroundOvarian cancer treatment includes cytoreductive surgery, platinum-based chemotherapy, and often poly (ADP-ribose) polymerase (PARP) inhibitors. Homologous recombination (HR)-deficiency is a well-established predictor of therapy sensitivity. However, over 50% of HR-proficient tumors also exhibit sensitivity to standard-of-care treatments. Currently, there are no biomarkers to identify which HR-proficient tumors will be sensitive to standard-of-care therapy. Replication stress may serve as a key determinant of response.

MethodsWe evaluated phospho-RPA2-T21 (pRPA2) foci via immunofluorescence as a potential biomarker of replication stress in formalin-fixed, paraffin-embedded tumor samples collected at diagnosis from patients treated with platinum chemotherapy (discovery cohort: n = 31, validation cohort: n = 244) or PARP inhibitors (n = 87). Recurrent tumors (n = 37) were also analyzed. pRPA2 scores were calculated using automated imaging analysis. Samples were defined as pRPA2-High if > 16% of cells had [&ge;] 2 pRPA2 foci.

ResultsIn the discovery cohort, HR-proficient, pRPA2-High tumors demonstrated significantly higher rates of pathologic complete response to platinum chemotherapy than HR-proficient, pRPA2-Low tumors. In the validation cohort, patients with HR-proficient, pRPA2-High tumors had significantly longer survival after platinum treatment than those with HR-proficient, pRPA2-Low tumors. Additionally, the pRPA2 assay effectively predicted survival outcomes in patients treated with PARP inhibitors and in recurrent tumor samples.

ConclusionOur study underscores the importance of considering replication stress markers alongside HR status in therapeutic planning. Our work suggest that this assay could be used throughout a patients treatment course to expand the number of patients receiving effective therapy while reducing unnecessary toxicity.
]]></description>
<dc:creator>Schab, A.</dc:creator>
<dc:creator>Compadre, A.</dc:creator>
<dc:creator>Drexler, R.</dc:creator>
<dc:creator>Loeb, M.</dc:creator>
<dc:creator>Rodriguez, K.</dc:creator>
<dc:creator>Brill, J.</dc:creator>
<dc:creator>Harrington, S.</dc:creator>
<dc:creator>Sandoval, C.</dc:creator>
<dc:creator>Sanders, B.</dc:creator>
<dc:creator>Kuroki, L.</dc:creator>
<dc:creator>McCourt, C.</dc:creator>
<dc:creator>Hagemann, A. R.</dc:creator>
<dc:creator>Thaker, P.</dc:creator>
<dc:creator>Mutch, D.</dc:creator>
<dc:creator>Powell, M.</dc:creator>
<dc:creator>Serra, V.</dc:creator>
<dc:creator>Hagemann, I. S.</dc:creator>
<dc:creator>Walts, A. E.</dc:creator>
<dc:creator>Karlan, B. Y.</dc:creator>
<dc:creator>Orsulic, S.</dc:creator>
<dc:creator>Fuh, K.</dc:creator>
<dc:creator>Sun, L.</dc:creator>
<dc:creator>Verma, P.</dc:creator>
<dc:creator>Lomonosova, E.</dc:creator>
<dc:creator>Zhao, P.</dc:creator>
<dc:creator>Khabele, D.</dc:creator>
<dc:creator>Mullen, M. M.</dc:creator>
<dc:date>2024-11-25</dc:date>
<dc:identifier>doi:10.1101/2024.11.21.624682</dc:identifier>
<dc:title><![CDATA[Replication stress marker phospho-RPA2 predicts response to platinum and PARP inhibitors in homologous recombination-proficient ovarian cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.22.624784v1?rss=1">
<title>
<![CDATA[
Unraveling SARS-CoV-2 Host-Response Heterogeneity through Longitudinal Molecular Subtyping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.22.624784v1?rss=1</link>
<description><![CDATA[
Hospitalized COVID-19 patients exhibit diverse immune responses during acute infection, which are associated with a wide range of clinical outcomes. However, understanding these immune heterogeneities and their links to various clinical complications, especially long COVID, remains a challenge. In this study, we performed unsupervised subtyping of longitudinal multi-omics immunophenotyping in over 1,000 hospitalized patients, identifying two critical subtypes linked to mortality or mechanical ventilation with prolonged hospital stay and three severe subtypes associated with timely acute recovery. We confirmed that unresolved systemic inflammation and T-cell dysfunctions were hallmarks of increased severity and further distinguished patients with similar acute respiratory severity by their distinct immune profiles, which correlated with differences in demographic and clinical complications. Notably, one critical subtype (SubF) was uniquely characterized by early excessive inflammation, insufficient anticoagulation, and fatty acid dysregulation, alongside higher incidences of hematologic, cardiac, and renal complications, and an elevated risk of long COVID. Among the severe subtypes, significant differences in viral clearance and early antiviral responses were observed, with one subtype (SubC) showing strong early T-cell cytotoxicity but a poor humoral response, slower viral clearance, and greater risks of chronic organ dysfunction and long COVID. These findings provide crucial insights into the complex and context-dependent nature of COVID-19 immune responses, highlighting the importance of personalized therapeutic strategies to improve both acute and long-term outcomes.
]]></description>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Nie, Y.</dc:creator>
<dc:creator>Maguire, C.</dc:creator>
<dc:creator>Syphurs, C.</dc:creator>
<dc:creator>Sheen, H.</dc:creator>
<dc:creator>Karoly, M.</dc:creator>
<dc:creator>Lapp, L.</dc:creator>
<dc:creator>Gygi, J. P.</dc:creator>
<dc:creator>Jayavelu, N. D.</dc:creator>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Hoch, A.</dc:creator>
<dc:creator>IMPACC Network,</dc:creator>
<dc:creator>Corry, D.</dc:creator>
<dc:creator>Kheradmand, F.</dc:creator>
<dc:creator>McComsey, G. A.</dc:creator>
<dc:creator>Fernandez-Sesma, A.</dc:creator>
<dc:creator>Simon, V.</dc:creator>
<dc:creator>Metcalf, J. P.</dc:creator>
<dc:creator>Agudelo Higuita, N. I.</dc:creator>
<dc:creator>Messer, W. B.</dc:creator>
<dc:creator>David, M. M.</dc:creator>
<dc:creator>Nadeau, K. C.</dc:creator>
<dc:creator>Kraft, M.</dc:creator>
<dc:creator>Bime, C.</dc:creator>
<dc:creator>Schaenman, J.</dc:creator>
<dc:creator>Erle, D.</dc:creator>
<dc:creator>Calfee, C. S.</dc:creator>
<dc:creator>Atkinson, M. A.</dc:creator>
<dc:creator>Brackenridge, S. C.</dc:creator>
<dc:creator>Hafler, D. A.</dc:creator>
<dc:creator>Shaw, A. C.</dc:creator>
<dc:creator>Rahman, A.</dc:creator>
<dc:creator>Hough, C. L.</dc:creator>
<dc:creator>Geng, L. N.</dc:creator>
<dc:creator>Ozonoff, A.</dc:creator>
<dc:creator>Haddad, E. K.</dc:creator>
<dc:creator>Reed, E. F.</dc:creator>
<dc:creator>Bakel, H. v.</dc:creator>
<dc:creator>Kim-Schulze, S. H.</dc:creator>
<dc:creator>Krammer, F.</dc:creator>
<dc:creator>Wilson, M.</dc:creator>
<dc:creator>Eckalbar, W.</dc:creator>
<dc:creator>Bosinger,</dc:creator>
<dc:date>2024-11-25</dc:date>
<dc:identifier>doi:10.1101/2024.11.22.624784</dc:identifier>
<dc:title><![CDATA[Unraveling SARS-CoV-2 Host-Response Heterogeneity through Longitudinal Molecular Subtyping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.25.625195v1?rss=1">
<title>
<![CDATA[
CRISPRa-mediated disentanglement of the Dux-MERVL axis in the 2C-like state, totipotency and cell death 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.25.625195v1?rss=1</link>
<description><![CDATA[
Transposable elements (TEs) provide sequences that are powerful cis-regulatory drivers of gene expression programmes. This is particularly apparent during early development when many TEs become de-repressed. MERVL elements are highly yet transiently upregulated in mouse totipotent 2-cell (2C) embryos during major zygotic genome activation (ZGA), and in 2C-like cells in vitro. One of the most powerful activators of MERVL is the pioneer transcription factor, Dux. However, apparent differences lie in the requirement for Dux versus MERVL activation in embryos, for unclear reasons. Moreover, sustained Dux activation causes cell toxicity in multiple cell types, which may or may not be linked to MERVL activation. Using a CRISPR-activation, 2C-GFP reporter system, we have unpicked the relative role of Dux and MERVL in ZGA, totipotent-like characteristics and cell toxicity. We find that direct MERVL activation comprises only a portion of the Dux-dependent transcriptome, and which is sufficient for expanded fate potential, but not other totipotency features. Conversely, Dux-induced pathology is independent of MERVL activation and involves induction of the pro-apoptotic factor, Noxa. Our study highlights the complexity of the Dux-MERVL transcriptional network and uncovers a new player in Dux-driven pathology.
]]></description>
<dc:creator>Chammas, P.</dc:creator>
<dc:creator>Xie, S. Q.</dc:creator>
<dc:creator>Sepulveda-Rincon, L. P.</dc:creator>
<dc:creator>Leeke, B. J.</dc:creator>
<dc:creator>Dore, M. H.</dc:creator>
<dc:creator>Dormann, D.</dc:creator>
<dc:creator>Wagner, R. T.</dc:creator>
<dc:creator>McManus, M. T.</dc:creator>
<dc:creator>Karimi, M. M.</dc:creator>
<dc:creator>Young, G.</dc:creator>
<dc:creator>Percharde, M.</dc:creator>
<dc:date>2024-11-25</dc:date>
<dc:identifier>doi:10.1101/2024.11.25.625195</dc:identifier>
<dc:title><![CDATA[CRISPRa-mediated disentanglement of the Dux-MERVL axis in the 2C-like state, totipotency and cell death]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.26.625482v1?rss=1">
<title>
<![CDATA[
A contextual fear conditioning paradigm in head-fixed mice exploring virtual reality. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625482v1?rss=1</link>
<description><![CDATA[
Contextual fear conditioning is a classical laboratory task that tests associative memory formation and recall. Techniques such as multi-photon microscopy and holographic stimulation offer tremendous opportunities to understand the neural underpinnings of these memories. However, these techniques generally require animals to be head-fixed. There are few paradigms that test contextual fear conditioning in head-fixed mice, and none where the behavioral outcome following fear conditioning is freezing, the most common measure of fear in freely moving animals. To address this gap, we developed a contextual fear conditioning paradigm in head-fixed mice using virtual reality (VR) environments. We designed an apparatus to deliver tail shocks (unconditioned stimulus, US) while mice navigated a VR environment (conditioned stimulus, CS). The acquisition of contextual fear was tested when the mice were reintroduced to the shock-paired VR environment the following day. We tested three different versions of this paradigm and, in all of them, observed an increased conditioned fear response characterized by increased freezing behavior. This was especially prominent during the first trial in the shock-paired VR environment, compared to a neutral environment where the mice received no shocks. Our results demonstrate that head-fixed mice can be fear conditioned in VR, discriminate between a feared and neutral VR context, and display freezing as a conditioned response, similar to freely behaving animals. Furthermore, using a two-photon microscope, we imaged from large populations of hippocampal CA1 neurons before, during, and following contextual fear conditioning. Our findings reconfirmed those from the literature on freely moving animals, showing that CA1 place cells undergo remapping and show narrower place fields following fear conditioning. Our approach offers new opportunities to study the neural mechanisms underlying the formation, recall, and extinction of contextual fear memories. As the head-fixed preparation is compatible with multi-photon microscopy and holographic stimulation, it enables long-term tracking and manipulation of cells throughout distinct memory stages and provides subcellular resolution for investigating axonal, dendritic, and synaptic dynamics in real-time.
]]></description>
<dc:creator>Krishnan, S.</dc:creator>
<dc:creator>Dong, C.</dc:creator>
<dc:creator>Ratigan, H.</dc:creator>
<dc:creator>Morales-Rodriguez, D.</dc:creator>
<dc:creator>Cherian, C.</dc:creator>
<dc:creator>Sheffield, M.</dc:creator>
<dc:date>2024-11-27</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625482</dc:identifier>
<dc:title><![CDATA[A contextual fear conditioning paradigm in head-fixed mice exploring virtual reality.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.26.625496v1?rss=1">
<title>
<![CDATA[
Programmable epigenome editing by transient delivery of CRISPR epigenome editor ribonucleoproteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625496v1?rss=1</link>
<description><![CDATA[
Programmable epigenome editors modify gene expression in mammalian cells by altering the local chromatin environment at target loci without inducing DNA breaks. However, the large size of CRISPR-based epigenome editors poses a challenge to their broad use in biomedical research and as future therapies. Here, we present Robust ENveloped Delivery of Epigenome-editor Ribonucleoproteins (RENDER) for transiently delivering programmable epigenetic repressors (CRISPRi, DNMT3A-3L-dCas9, CRISPRoff) and activator (TET1-dCas9) as ribonucleoprotein complexes into human cells to modulate gene expression. After rational engineering, we show that RENDER induces durable epigenetic silencing of endogenous genes across various human cell types, including primary T cells. Additionally, we apply RENDER to epigenetically repress endogenous genes in human stem cell-derived neurons, including the reduction of the neurodegenerative disease associated V337M-mutated Tau protein. Together, our RENDER platform advances the delivery of CRISPR-based epigenome editors into human cells, broadening the use of epigenome editing in fundamental research and therapeutic applications.
]]></description>
<dc:creator>Nunez, J. K.</dc:creator>
<dc:creator>Xu, D.</dc:creator>
<dc:creator>Besselink, S.</dc:creator>
<dc:creator>Ramadoss, G. N.</dc:creator>
<dc:creator>Dierks, P. H.</dc:creator>
<dc:creator>Lubin, J. P.</dc:creator>
<dc:creator>Pattali, R. K.</dc:creator>
<dc:creator>Brim, J. I.</dc:creator>
<dc:creator>Christenson, A. E.</dc:creator>
<dc:creator>Colias, P. J.</dc:creator>
<dc:creator>Ornelas, I. J.</dc:creator>
<dc:creator>Nguyen, C. D.</dc:creator>
<dc:creator>Chasins, S. E.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:date>2024-11-27</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625496</dc:identifier>
<dc:title><![CDATA[Programmable epigenome editing by transient delivery of CRISPR epigenome editor ribonucleoproteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.25.622742v1?rss=1">
<title>
<![CDATA[
Arterial endothelial deletion of Alk1 causes epistaxis and cerebral microhemorrhage with aberrant arteries and defective smooth muscle coverage. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.25.622742v1?rss=1</link>
<description><![CDATA[
Hereditary Hemorrhagic Telangiectasia (HHT) is an autosomal dominant vascular disorder with manifestations including severe nose bleeding and microhemorrhage in brains. Despite being the second most common inherited bleeding disorder, the pathophysiological mechanism underlying HHT-associated hemorrhage is poorly understood. HHT pathogenesis is thought to follow a Knudsonian two-hit model, requiring a second somatic mutation for lesion formation. Mutations in activin receptor-like kinase 1 (ALK1) gene cause HHT type 2. We hypothesize that somatic mutation of Alk1 in arterial endothelial cells (AECs) leads to arterial defects and hemorrhage. Here, we mutated Alk1 in AECs in postnatal mice using Bmx(PAC)-CreERT2 and found that somatic arterial endothelial mutation of Alk1 was sufficient to induce spontaneous epistaxis and multifocal cerebral microhemorrhage. This bleeding occurred in the presence of tortuous and enlarged blood vessels, loss of arterial molecular marker Efnb2, disorganization of vascular smooth muscle, and impaired vasoregulation. Our data suggest that arterial endothelial deletion of Alk1 leading to reduced arterial identity and disrupted vascular smooth muscle cell coverage is a plausible molecular mechanism for HHT-associated severe epistaxis. This work provides the first evidence that somatic Alk1 mutation in AECs can cause hemorrhagic vascular lesions, offering a novel preclinical model critically needed for studying HHT-associated epistaxis, and delineating an arterial mechanism to HHT pathophysiology.
]]></description>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Jacobs, K. A.</dc:creator>
<dc:creator>Raygor, K. P.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Wang, R. A.</dc:creator>
<dc:date>2024-11-28</dc:date>
<dc:identifier>doi:10.1101/2024.11.25.622742</dc:identifier>
<dc:title><![CDATA[Arterial endothelial deletion of Alk1 causes epistaxis and cerebral microhemorrhage with aberrant arteries and defective smooth muscle coverage.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.26.625329v1?rss=1">
<title>
<![CDATA[
An updated reference genome sequence and annotation reveals gene losses and gains underlying naked mole-rat biology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625329v1?rss=1</link>
<description><![CDATA[
The naked mole-rat (NMR; Heterocephalus glaber) is a eusocial subterranean rodent with a highly unusual set of physiological traits that has attracted great interest amongst the scientific community. However, the genetic basis of most of these traits has not been elucidated. To facilitate our understanding of the molecular mechanisms underlying NMR physiology and behaviour, we generated a long-read chromosomal-level genome assembly of the NMR. This genome was subsequently annotated and incorporated into multiple whole genome alignments in the Ensembl database. Our long-read assembly identified thousands of repeats and genes that were previously unassembled in the NMR and improved the results of routinely used short-read sequencing-based experiments such as RNA-seq, snRNA-seq, and ATAC-seq. We identified several spermatozoa related gene losses that may underlie the unique degenerative sperm phenotype in NMRs (IRGC, FSCB, AKAP3, MROH2B, CATSPER1, DCDC2C, ATP1A4, TEKT5, and ZAN), and an additional gene loss related to the established NK-cell absence in NMRs (PILRB). We resolved several tandem duplications in genes related to pathways underlying unique NMR adaptations including hypoxia tolerance, oxidative stress, and nervous system protection (TINF2, TCP1, KYAT1). Lastly, we describe our ongoing efforts to generate a reference telomere-to-telomere assembly in the NMR which includes the resolution of complex gene families. This new reference genome should accelerate the discovery of the genetic underpinnings of NMR physiology and adaptation.
]]></description>
<dc:creator>Sokolowski, D. J.</dc:creator>
<dc:creator>Miclaus, M.</dc:creator>
<dc:creator>Nater, A.</dc:creator>
<dc:creator>Faykoo-Martinez, M.</dc:creator>
<dc:creator>Hoekzema, K.</dc:creator>
<dc:creator>Zuzarte, P.</dc:creator>
<dc:creator>Monis, S.</dc:creator>
<dc:creator>Alvi, S. A.</dc:creator>
<dc:creator>Erdmann, J.</dc:creator>
<dc:creator>Erdmann, A. L.</dc:creator>
<dc:creator>Kumaragurubaran, R.</dc:creator>
<dc:creator>Bayerl, J.</dc:creator>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Karimpour, N.</dc:creator>
<dc:creator>Ungerleider, K.</dc:creator>
<dc:creator>Hou, H.</dc:creator>
<dc:creator>Fergal, M.</dc:creator>
<dc:creator>Hourlier, T.</dc:creator>
<dc:creator>Clarke, Z. A.</dc:creator>
<dc:creator>Lischer, H. E.</dc:creator>
<dc:creator>Leordean, D. V.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Pugh, T. J.</dc:creator>
<dc:creator>Smith, E. S. J.</dc:creator>
<dc:creator>Haggerty, L.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:creator>Lilue, J.</dc:creator>
<dc:creator>Holmes, M. M.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:creator>Bruggmann, R.</dc:creator>
<dc:creator>Simpson, J. T.</dc:creator>
<dc:creator>Balmus, G.</dc:creator>
<dc:creator>Wilson, M. D.</dc:creator>
<dc:date>2024-11-28</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625329</dc:identifier>
<dc:title><![CDATA[An updated reference genome sequence and annotation reveals gene losses and gains underlying naked mole-rat biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.26.625540v1?rss=1">
<title>
<![CDATA[
Actin filament assembly driven by distributive polymerases clustered on membrane surfaces 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625540v1?rss=1</link>
<description><![CDATA[
Actin filaments created by the Arp2/3 complex form branched networks, that grow and push against cellular membranes. We employ theory and simulation to describe how membrane surfaces accelerate filament assembly via clustering of proteins that bind actin monomers and/or profilin-actin complexes. Briefly, thermal fluctuations drive filament tips on constrained, two-dimensional random walks across the membrane, where they encounter multiple actin-charged polymerases. At low actin concentrations, filament elongation is limited by delivery of monomers to the membrane surface; at high actin concentrations, elongation depends on how quickly fluctuating filaments search the membrane. Using experimentally measured parameter values we conclude that surface-mediated polymerization can outpace solution-mediated elongation, even at high actin concentrations (>200 {micro}M). The finite time required for profilin dissociation decreases the advantage conferred by surface-associated polymerases, but only in the absence of force. Load forces enhance the effect of surface polymerases, which can both accelerate elongation and increase the force required to stall filament assembly.
]]></description>
<dc:creator>Mullins, D.</dc:creator>
<dc:creator>Skruber, K.</dc:creator>
<dc:date>2024-11-28</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625540</dc:identifier>
<dc:title><![CDATA[Actin filament assembly driven by distributive polymerases clustered on membrane surfaces]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.27.625786v1?rss=1">
<title>
<![CDATA[
Genome Organization with CUT and Tag (GO-CaT) identifies substructure and maturation of lamina-associated domains in neurons of the developing human brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.27.625786v1?rss=1</link>
<description><![CDATA[
Approximately 30-40% of the human genome is anchored to the nuclear lamina (NL) through variably sized (10 kb-10 Mb) lamina-associated domains (LADs), which can be classified into two subtypes (T1 and T2) based on their level of lamina-association. The dynamics of LAD substructure in cells that remain postmitotic for long periods of time are poorly understood. Here, we developed Genome Organization with CUT and Tag (GO-CaT) to determine the T1- and T2-LAD substructure of postmitotic excitatory neurons isolated from the prenatal and adult human cortex. While T1-LADs exhibited epigenomic features characteristic of stable, cell type-invariant LADs including strong transcriptional repression, in prenatal neurons, T2-LADs were enriched for promoter-enhancer DNA interactions, intermediate levels of gene expression, and genetic risk associated with neurodevelopmental and cognitive disorders. In adult cortical neurons, T1-LADs were expanded in size and genomic coverage, incorporating the majority of the prenatal T2-LADs, sequestering genes involved in neurodevelopment. In contrast, the minority of prenatal T2-LADs that relocated to inter-LAD regions in adult neurons were enriched for processes related to synaptic function. Overall, these data provide evidence that LADs "mature" in postmitotic neurons, remodeling from a genomic architecture that is more permissive for the dynamics of transcription of development to one that is more restricted and focused on the decades-long transcriptional needs of adult brain neurons.
]]></description>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Gil, E.</dc:creator>
<dc:creator>Ahanger, S. H.</dc:creator>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:creator>Lim, D.</dc:creator>
<dc:date>2024-11-28</dc:date>
<dc:identifier>doi:10.1101/2024.11.27.625786</dc:identifier>
<dc:title><![CDATA[Genome Organization with CUT and Tag (GO-CaT) identifies substructure and maturation of lamina-associated domains in neurons of the developing human brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.26.625568v1?rss=1">
<title>
<![CDATA[
Structural and functional insights into activation and regulation of the dynein-dynactin-NuMA complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625568v1?rss=1</link>
<description><![CDATA[
During cell division, NuMA orchestrates the focusing of microtubule minus-ends in spindle poles and cortical force generation on astral microtubules by interacting with dynein motors, microtubules, and other cellular factors. Here, we used in vitro reconstitution, cryo-electron microscopy, and live cell imaging to understand the mechanism and regulation of NuMA. We determined the structure of the processive dynein/dynactin/NuMA complex (DDN) and showed that the NuMA N-terminus drives dynein motility in vitro and facilitates dynein-mediated transport in live cells. The C-terminus of NuMA directly binds to and suppresses the dynamics of the microtubule minus-end. Full-length NuMA is autoinhibited for its interactions with dynein and microtubules, but mitotically phosphorylated NuMA activates dynein in vitro and interphase cells. Together with dynein, activated full-length NuMA focuses microtubule minus-ends into aster-like structures. These results provide critical insights into the activation of NuMA and dynein for their mitotic functions.
]]></description>
<dc:creator>Aslan, M.</dc:creator>
<dc:creator>Cho, N. H.</dc:creator>
<dc:creator>d'Amico, E. A.</dc:creator>
<dc:creator>Taheri, A.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Zhong, X.</dc:creator>
<dc:creator>Blaauw, M.</dc:creator>
<dc:creator>Carter, A. P.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:creator>Yildiz, A.</dc:creator>
<dc:date>2024-11-29</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625568</dc:identifier>
<dc:title><![CDATA[Structural and functional insights into activation and regulation of the dynein-dynactin-NuMA complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.29.622360v1?rss=1">
<title>
<![CDATA[
NuMA mechanically reinforces the spindle independently of its partner dynein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.29.622360v1?rss=1</link>
<description><![CDATA[
Both motor and non-motor proteins organize microtubules to build the spindle and maintain it against opposing forces. NuMA, a long microtubule binding protein, is essential to spindle structure and function. NuMA recruits the motor dynein to spindle microtubule minus-ends to actively cluster them, but whether NuMA performs other spindle roles remains unknown. Here, we show that NuMA acts independently of dynein to passively reinforce the mammalian spindle. NuMA that cannot bind dynein is sufficient to protect spindle poles against fracture under external force. In contrast, NuMA with a shorter coiled-coil or disrupted self-interactions cannot protect spindle poles, and NuMA turnover differences cannot explain mechanical differences. In vitro, NuMAs C-terminus self-interacts and bundles microtubules without dynein, dependent on residues essential to pole protection in vivo. Together, this suggests that NuMA reinforces spindle poles by crosslinking microtubules, using its long coiled-coiled and self-interactions to reach multiple, far-reaching pole microtubules. We propose that NuMA acts as a mechanical "multitasker" targeting contractile motor activity and separately crosslinking microtubules, both functions synergizing to drive spindle mechanical robustness.
]]></description>
<dc:creator>Cho, N. H.</dc:creator>
<dc:creator>Aslan, M.</dc:creator>
<dc:creator>Yildiz, A.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2024-12-01</dc:date>
<dc:identifier>doi:10.1101/2024.11.29.622360</dc:identifier>
<dc:title><![CDATA[NuMA mechanically reinforces the spindle independently of its partner dynein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.01.626277v1?rss=1">
<title>
<![CDATA[
The GluA1 cytoplasmic tail regulates intracellular AMPA receptor trafficking and synaptic transmission onto dentate gyrus GABAergic interneurons, gating response to novelty 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.01.626277v1?rss=1</link>
<description><![CDATA[
The GluA1 subunit, encoded by the putative schizophrenia-associated gene GRIA1, is required for activity-regulated AMPA receptor (AMPAR) trafficking, and plays a key role in cognitive and affective function. The cytoplasmic, carboxy-terminal domain (CTD) is the most divergent region across AMPAR subunits. The GluA1 CTD has received considerable attention for its role during long-term potentiation (LTP) at CA1 pyramidal neuron synapses. However, its function at other synapses and, more broadly, its contribution to different GluA1-dependent processes, is poorly understood. Here, we used mice with a constitutive truncation of the GluA1 CTD to dissect its role regulating AMPAR localization and function as well as its contribution to cognitive and affective processes. We found that GluA1 CTD truncation affected AMPAR subunit levels and intracellular trafficking. {Delta}CTD GluA1 mice exhibited no memory deficits, but presented exacerbated novelty-induced hyperlocomotion and dentate gyrus granule cell (DG GC) hyperactivity, among other behavioral alterations. Mechanistically, we found that AMPAR EPSCs onto DG GABAergic interneurons were significantly reduced, presumably underlying, at least in part, the observed changes in neuronal activity and behavior. In summary, this study dissociates CTD-dependent from CTD-independent GluA1 functions, unveiling the GluA1 CTD as a crucial hub regulating AMPAR function in a cell type-specific manner.
]]></description>
<dc:creator>Leana-Sandoval, G.</dc:creator>
<dc:creator>Kolli, A. V.</dc:creator>
<dc:creator>Chinn, C. A.</dc:creator>
<dc:creator>Madrid, A.</dc:creator>
<dc:creator>Lo, I.</dc:creator>
<dc:creator>Sandoval, M. A.</dc:creator>
<dc:creator>Alizo Vera, V.</dc:creator>
<dc:creator>Simms, J.</dc:creator>
<dc:creator>Wood, M. A.</dc:creator>
<dc:creator>Diaz Alonso, J.</dc:creator>
<dc:date>2024-12-02</dc:date>
<dc:identifier>doi:10.1101/2024.12.01.626277</dc:identifier>
<dc:title><![CDATA[The GluA1 cytoplasmic tail regulates intracellular AMPA receptor trafficking and synaptic transmission onto dentate gyrus GABAergic interneurons, gating response to novelty]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.02.626379v1?rss=1">
<title>
<![CDATA[
The VGCC auxiliary subunit α2δ1 is an extracellular GluA1 interactor and regulates LTP, spatial memory, and seizure susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.02.626379v1?rss=1</link>
<description><![CDATA[
Activity-dependent synaptic accumulation of AMPA receptors (AMPARs) and subsequent long-term synaptic strengthening underlie different forms of learning and memory. The AMPAR subunit GluA1 amino-terminal domain is essential for synaptic docking of AMPAR during LTP, but the precise mechanisms involved are not fully understood. Using unbiased proteomics, we identified the epilepsy and intellectual disability-associated VGCC auxiliary subunit 2{delta}1 as a candidate extracellular AMPAR slot. Presynaptic 2{delta}1 deletion in CA3 affects synaptic AMPAR incorporation during long-term potentiation, but not basal synaptic transmission, at CA1 synapses. Consistently, mice lacking 2{delta}1 in CA3 display a specific impairment in CA1-dependent spatial memory, but not in memory tests involving other cortical regions. Decreased seizure susceptibility in mice lacking 2{delta}1 in CA3 suggests a regulation of circuit excitability by 2{delta}1/AMPAR interactions. Our study sheds light on the regulation of activity-dependent AMPAR trafficking, and highlights the synaptic organizing roles of 2{delta}1.

Significance statementActivity-dependent accumulation of AMPA receptors (AMPARs) at excitatory synapses and subsequent synaptic strengthening underlies long-term potentiation (LTP), forms of learning and memory, and some epilepsies. The "slot model" posits that postsynaptic scaffolding contain "slots" for AMPAR complexes, and that increased synaptic activity augments the availability of slots to accommodate more receptors, thereby strengthening synapses and enabling LTP. The presence of the GluA1 AMPAR subunit amino-terminal domain (ATD) has recently emerged as an additional requirement for LTP. Here we identify the auxiliary voltage-gated calcium channel subunit 2{delta}1 as a GluA1 ATD interacting protein and provide evidence supporting a role for 2{delta}1 as an extracellular AMPAR slot regulating activity dependent synaptic AMPAR clustering, excitability, and cognitive function.
]]></description>
<dc:creator>Leana-Sandoval, G.</dc:creator>
<dc:creator>Kolli, A. V.</dc:creator>
<dc:creator>Sandoval, M. A.</dc:creator>
<dc:creator>Saavedra, E. M.</dc:creator>
<dc:creator>Li, K. H.</dc:creator>
<dc:creator>Chen, L. Y.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Ramirez-Franco, J. J.</dc:creator>
<dc:creator>Diaz-Alonso, J.</dc:creator>
<dc:date>2024-12-02</dc:date>
<dc:identifier>doi:10.1101/2024.12.02.626379</dc:identifier>
<dc:title><![CDATA[The VGCC auxiliary subunit α2δ1 is an extracellular GluA1 interactor and regulates LTP, spatial memory, and seizure susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.27.625710v1?rss=1">
<title>
<![CDATA[
Polyfunctional IL-21+ IFNg+ T follicular helper cells contribute to checkpoint inhibitor diabetes mellitus and can be targeted by JAK inhibitor therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.27.625710v1?rss=1</link>
<description><![CDATA[
Immune checkpoint inhibitors (ICI) have revolutionized cancer therapy, but their use is limited by the development of autoimmunity in healthy tissues as a side effect of treatment. Such immune-related adverse events (IrAE) contribute to hospitalizations, cancer treatment interruption and even premature death. ICI-induced autoimmune diabetes mellitus (ICI-T1DM) is a life-threatening IrAE that presents with rapid pancreatic beta-islet cell destruction leading to hyperglycemia and life-long insulin dependence. While prior reports have focused on CD8+ T cells, the role for CD4+ T cells in ICI-T1DM is less understood. Here, we identify expansion CD4+ T follicular helper (Tfh) cells expressing interleukin 21 (IL-21) and interferon gamma (IFN{gamma}) as a hallmark of ICI-T1DM. Furthermore, we show that both IL-21 and IFN{gamma} are critical cytokines for autoimmune attack in ICI-T1DM. Because IL-21 and IFN{gamma} both signal through JAK-STAT pathways, we reasoned that JAK inhibitors (JAKi) may protect against ICI-T1DM. Indeed, JAKi provide robust in vivo protection against ICI-T1DM in a mouse model that is associated with decreased islet-infiltrating Tfh cells. Moreover, JAKi therapy impaired Tfh cell differentiation in patients with ICI-T1DM. These studies highlight CD4+ Tfh cells as underrecognized but critical mediators of ICI-T1DM that may be targeted with JAKi to prevent this grave IrAE.

VISUAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=92 SRC="FIGDIR/small/625710v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Huang, N.</dc:creator>
<dc:creator>Ortega, J.</dc:creator>
<dc:creator>Kimbrell, K.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Scott, L. N.</dc:creator>
<dc:creator>Peluso, E. M.</dc:creator>
<dc:creator>Wang, S. J.</dc:creator>
<dc:creator>Kao, E.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Olay, J.</dc:creator>
<dc:creator>Quandt, Z.</dc:creator>
<dc:creator>Angell, T. E.</dc:creator>
<dc:creator>Su, M. A.</dc:creator>
<dc:creator>Lechner, M. G.</dc:creator>
<dc:date>2024-12-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.27.625710</dc:identifier>
<dc:title><![CDATA[Polyfunctional IL-21+ IFNg+ T follicular helper cells contribute to checkpoint inhibitor diabetes mellitus and can be targeted by JAK inhibitor therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.27.625750v1?rss=1">
<title>
<![CDATA[
Coordinated variation in the immune systems and microbiomes of healthy humans is linked to tonic interferon states 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.27.625750v1?rss=1</link>
<description><![CDATA[
Human immune systems are highly variable, with most variation attributable to non-genetic sources. The gut microbiome crucially shapes the immune system; however, its relationship with the baseline immune states of healthy humans remains incompletely understood. Therefore, we performed multi-omic profiling of 110 healthy participants through the ImmunoMicrobiome study. A factor-based integrative approach identified coordinated variation, revealing that the tonic interferon response was amongst the most variable immune features in healthy participants. Microbiome composition, pathways, and stool metabolites varied concomitantly with interferon response pathways. Distinct transcriptional programs involving inflammation and TGF-{beta} in SIGLEC-1high monocytes and CD69high activated MAIT and NK cells were representative of these programs. Our study provides extensive data to examine the relationship between the immune states and microbiomes of healthy individuals at steady state, which paves the way for delineating inter-individual differences relevant for disease susceptibility and responses to therapy.
]]></description>
<dc:creator>Babdor, J.</dc:creator>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Davidson, B.</dc:creator>
<dc:creator>Noecker, C.</dc:creator>
<dc:creator>Rahim, M.</dc:creator>
<dc:creator>Bisanz, J. E.</dc:creator>
<dc:creator>Tenvooren, I.</dc:creator>
<dc:creator>Marquez, D.</dc:creator>
<dc:creator>Calvo, M.</dc:creator>
<dc:creator>Johri, V.</dc:creator>
<dc:creator>McCarthy, E.</dc:creator>
<dc:creator>Shaheed, A.</dc:creator>
<dc:creator>Ekstrand, C.</dc:creator>
<dc:creator>Weakley, A. M.</dc:creator>
<dc:creator>Yu, F. B.</dc:creator>
<dc:creator>Fiehn, O.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:date>2024-12-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.27.625750</dc:identifier>
<dc:title><![CDATA[Coordinated variation in the immune systems and microbiomes of healthy humans is linked to tonic interferon states]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.27.625761v1?rss=1">
<title>
<![CDATA[
Nucleotide GPT: Sequence-Based Deep Learning Prediction of Nuclear Subcompartment-Associated Genome Architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.27.625761v1?rss=1</link>
<description><![CDATA[
The spatial organization of the genome within the nucleus is partially determined by its interactions with distinct nuclear subcompartments, such as the nuclear lamina and nuclear speckles, which play key roles in gene regulation during development. However, whether these genome-nuclear subcompartment interactions are encoded in the underlying DNA sequence remains poorly understood. The mechanisms for gene regulation are primarily encoded in noncoding DNA sequences, but deciphering how these sequence features control gene expression remains a significant challenge in genomics. Here, we present Nucleotide GPT, a transformer-based model that predicts genomic associations with spatially distinct, physical nuclear subcompartments from DNA sequence alone. Pre-trained on a diverse set of multi-species genomes, we demonstrate Nucleotide GPTs genomic understanding through evaluation on diverse prediction tasks, including histone modifications, promoter detection, and transcription factor binding sites. When finetuned to predict genome interactions with two separate nuclear subcompartments - the lamina of the inner nuclear membrane and nuclear speckles that lie more interior - Nucleotide GPT achieves an average accuracy of 73.6% for lamina-associated domains (LADs) and 79.4% accuracy for speckle-associated domains (SPADs), averaged across three cortical development cell types. Analysis of the models learned representations through Uniform Manifold Approximation and Projection (UMAP) reveals that Nucleotide GPT develops internal embeddings that effectively distinguish LADs from inter-LADs, with predicted probabilities closely corresponding to experimentally determined LAD classifications. When examining these representations in the context of cell type-invariant constitutive LADs (cLADs) compared to cell type-specific LADs, the model assigns lower confidence scores to cell type-specific LADs compared to cLADs that are conserved across neuronal differentiation, suggesting sequence features may play a stronger role in maintaining cLAD associations. Examination of the models attention patterns at correctly classified regions suggests that specific sequence elements govern model decision making about nuclear subcompartment associations. Our results demonstrate the utility of transformer architectures for studying three-dimensional (3D) genome organization and substantiate a role for DNA sequence in determining nuclear subcompartment associations.
]]></description>
<dc:creator>Mclaughlin, S. M.</dc:creator>
<dc:creator>Ahanger, S. H.</dc:creator>
<dc:creator>Lim, D. A.</dc:creator>
<dc:date>2024-12-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.27.625761</dc:identifier>
<dc:title><![CDATA[Nucleotide GPT: Sequence-Based Deep Learning Prediction of Nuclear Subcompartment-Associated Genome Architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.29.626078v1?rss=1">
<title>
<![CDATA[
Rebuilding the autoimmune-damaged corneal stroma through topical lubrication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.29.626078v1?rss=1</link>
<description><![CDATA[
Corneal lubrication is the most common treatment for relieving the signs and symptoms of dry eye and is considered to be largely palliative with no regenerative functions. Here we challenge this notion by demonstrating that wetting the desiccated cornea of an aqueous-deficient mouse model with the simplest form of lubrication, a saline-based solution, is sufficient to rescue the severely disrupted collagen-rich architecture of the stroma, the largest corneal compartment that is essential to transparency and vision. At the single cell level we show that stromal keratocytes responsible for maintaining stromal integrity are converted from an inflammatory state into unique reparative cell states by lubrication alone, thus revealing the extensive plasticity of these cells and the regenerative function of lubricating the surface. We further show that the generation of a reparative phenotype is due, in part, to disruption of an IL1{beta} autocrine amplification loop promoting chronic inflammation. Thus, our study uncovers the regenerative potential of topical lubrication in dry eye and represents a paradigm shift in our understanding of its therapeutic impact.
]]></description>
<dc:creator>Efraim, Y.</dc:creator>
<dc:creator>Chen, F. Y. T.</dc:creator>
<dc:creator>Niknezhad, S. V.</dc:creator>
<dc:creator>Pham, D.</dc:creator>
<dc:creator>Cheong, K. N.</dc:creator>
<dc:creator>An, L.</dc:creator>
<dc:creator>Sinada, H.</dc:creator>
<dc:creator>McNamara, N. A.</dc:creator>
<dc:creator>Knox, S.</dc:creator>
<dc:date>2024-12-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.29.626078</dc:identifier>
<dc:title><![CDATA[Rebuilding the autoimmune-damaged corneal stroma through topical lubrication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.03.626676v1?rss=1">
<title>
<![CDATA[
Widespread gene-environment interactions shape the immune response to SARS-CoV-2 infection in hospitalized COVID-19 patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.03.626676v1?rss=1</link>
<description><![CDATA[
Genome-wide association studies performed in patients with coronavirus disease 2019 (COVID-19) have uncovered various loci significantly associated with susceptibility to SARS-CoV-2 infection and COVID-19 disease severity. However, the underlying cis-regulatory genetic factors that contribute to heterogeneity in the response to SARS-CoV-2 infection and their impact on clinical phenotypes remain enigmatic. Here, we used single-cell RNA-sequencing to quantify genetic contributions to cis-regulatory variation in 361,119 peripheral blood mononuclear cells across 63 COVID-19 patients during acute infection, 39 samples collected in the convalescent phase, and 106 healthy controls. Expression quantitative trait loci (eQTL) mapping across cell types within each disease state group revealed thousands of cis-associated variants, of which hundreds were detected exclusively in immune cells derived from acute COVID-19 patients. Patient-specific genetic effects dissipated as infection resolved, suggesting that distinct gene regulatory networks are at play in the active infection state. Further, 17.2% of tested loci demonstrated significant cell state interactions with genotype, with pathways related to interferon responses and oxidative phosphorylation showing pronounced cell state-dependent variation, predominantly in CD14+ monocytes. Overall, we estimate that 25.6% of tested genes exhibit gene-environment interaction effects, highlighting the importance of environmental modifiers in the transcriptional regulation of the immune response to SARS-CoV-2. Our findings underscore the importance of expanding the study of regulatory variation to relevant cell types and disease contexts and argue for the existence of extensive gene-environment effects among patients responding to an infection.
]]></description>
<dc:creator>Randolph, H. E.</dc:creator>
<dc:creator>Aguirre-Gamboa, R.</dc:creator>
<dc:creator>Brunet-Ratnasingham, E.</dc:creator>
<dc:creator>Nakanishi, T.</dc:creator>
<dc:creator>Locher, V.</dc:creator>
<dc:creator>Ketter, E.</dc:creator>
<dc:creator>Brandolino, C.</dc:creator>
<dc:creator>Larochelle, C.</dc:creator>
<dc:creator>Prat, A.</dc:creator>
<dc:creator>Arbour, N.</dc:creator>
<dc:creator>Dumaine, A.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:creator>Durand, M.</dc:creator>
<dc:creator>Richards, J. B.</dc:creator>
<dc:creator>Kaufmann, D. E.</dc:creator>
<dc:creator>Barreiro, L. B.</dc:creator>
<dc:date>2024-12-04</dc:date>
<dc:identifier>doi:10.1101/2024.12.03.626676</dc:identifier>
<dc:title><![CDATA[Widespread gene-environment interactions shape the immune response to SARS-CoV-2 infection in hospitalized COVID-19 patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.02.626017v1?rss=1">
<title>
<![CDATA[
Epigenetic investigation of multifocal small intestinal neuroendocrine tumours reveals accelerated ageing of tumours and epigenetic alteration of metabolic genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.02.626017v1?rss=1</link>
<description><![CDATA[
BackgroundSmall intestinal neuroendocrine tumours (SI-NETs) are the most common malignancy of the small intestine and around 50% of patients present in clinic with multifocal disease. Recent investigations into the genomic architecture of multifocal SI-NETs have found evidence that these synchronous primary tumours evolve independently of each other. They also have extremely low mutational burden and few known driver genes, suggesting that epigenetic dysregulation may be driving tumorigenesis. Very little is known about epigenetic gene regulation, metabolism and ageing in these tumours, and how these traits differ across multiple tumours within individual patients.

MethodsIn this study, we performed the first investigation of genome-wide DNA methylation in multifocal SI-NETs, assessing multiple primary tumours within each patient (n=79 primary tumours from 14 patients) alongside matched metastatic tumours (n=12) and normal intestinal epithelial tissue (n=9). We assessed multifocal SI-NET differential methylation using a novel method, comparing primary tumours with matched normal epithelial tissue and an enterochromaffin-enriched cell line to enrich for tumour-specific effects. This method reduced the identification of  false positive methylation differences driven by cell composition differences between tumour and normal epithelial tissue. We also assessed tumour ageing using epigenetic clocks and applied metabolic predictors in the dataset to assess methylation variation across key metabolic genes.

ResultsWe have identified 12,392 tumour-specific differentially methylated positions (Bonferroni corrected p<0.05) which were enriched for neural pathways. The expression levels of the genes associated with top sites were also found to be significantly altered in SI-NETs. Age acceleration was observed across SI-NETs and a variability in epigenetic  age of tumours within each patient, which we believe is reflecting the  order in which tumours have developed. This is supported by the correlation of age acceleration with somatic mutational count in the tumours. We have identified SI-NET associated alterations to the methylation patterns in key metabolic genes compared to matched normal tissue, which is more pronounced in metastatic tumours and tumours harbouring chromosome 18 loss of heterozygosity, indicating metabolic differences in these tumour subtypes.

ConclusionsWe have identified accelerated ageing and changes to regulation of metabolic genes, alongside an epigenetic signature of multifocal SI-NETs. These findings add to our understanding of multifocal SI-NET biology and their molecular differences which may be instrumental in the development of these elusive tumours.
]]></description>
<dc:creator>Webster, A. P.</dc:creator>
<dc:creator>Makinen, N.</dc:creator>
<dc:creator>Mensah, N.</dc:creator>
<dc:creator>Castignani, C.</dc:creator>
<dc:creator>Larose Cadieux, E.</dc:creator>
<dc:creator>Shivdasani, R.</dc:creator>
<dc:creator>Singh, P.</dc:creator>
<dc:creator>Vaikkinen, H.</dc:creator>
<dc:creator>Dhami, P.</dc:creator>
<dc:creator>Ecker, S.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:creator>Rimmer, B.</dc:creator>
<dc:creator>Henderson, S.</dc:creator>
<dc:creator>Herrero, J.</dc:creator>
<dc:creator>Suderman, M.</dc:creator>
<dc:creator>Yousefi, P. D.</dc:creator>
<dc:creator>Beck, S.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Nakakura, E.</dc:creator>
<dc:creator>Thirlwell, C.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.02.626017</dc:identifier>
<dc:title><![CDATA[Epigenetic investigation of multifocal small intestinal neuroendocrine tumours reveals accelerated ageing of tumours and epigenetic alteration of metabolic genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.02.626435v1?rss=1">
<title>
<![CDATA[
Prediabetes Phenotype Clusters in the Diabetes Prevention Program Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.02.626435v1?rss=1</link>
<description><![CDATA[
ObjectiveThe purpose of this study was to apply clustering methods to identify and characterize prediabetes phenotypes and their relationships with treatment arm and type 2 diabetes (T2D) outcomes in the Diabetes Prevention Program (DPP), and to compare the utility of additional clustering measures in phenotype characterization and T2D risk stratification.

Research Design and MethodsThis was a secondary analysis of data from a subset of participants (n=994) from the previously completed Diabetes Prevention Program trial. Unsupervised k-means clustering analysis was applied to derive the optimal number of clusters of participants based on common clinical risk factors alone or common risk factors plus more comprehensive measures of glucose tolerance and body composition.

ResultsFive clusters were derived from common clinical characteristics and the addition of comprehensive measures of glucose tolerance and body composition. Within each modeling approach, participants show significantly different levels of risk factors. The clinical only model showed higher accuracy for time to T2D, however the more comprehensive models further differentiated a metabolically health overweight phenotype. For both models, the greatest differentiation in determining time to T2D was in the metformin arm of the trial.

ConclusionsData driven clustering of patients with prediabetes allows for identification of prediabetes phenotypes at greater risk for disease progression and responses to risk reduction interventions. Further investigation into phenotypic differences in treatment response could enable better personalization of prediabetes and T2D prevention and treatment choices.
]]></description>
<dc:creator>Stroebel, B. M.</dc:creator>
<dc:creator>Gadgil, M.</dc:creator>
<dc:creator>Lewis, K.</dc:creator>
<dc:creator>Longoria, K.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Flowers, E.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.02.626435</dc:identifier>
<dc:title><![CDATA[Prediabetes Phenotype Clusters in the Diabetes Prevention Program Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.02.626490v1?rss=1">
<title>
<![CDATA[
Target cell tension regulates macrophage trogocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.02.626490v1?rss=1</link>
<description><![CDATA[
Macrophages are known to engulf small membrane fragments, or trogocytose, target cells and pathogens, rather than fully phagocytose them. However, little is known about what causes macrophages to choose trogocytosis versus phagocytosis. Here, we report that cortical tension of target cells is a key regulator of macrophage trogocytosis. At low tension, macrophages will preferentially trogocytose antibody-opsonized cells, while at high tension they tend towards phagocytosis. Using model vesicles, we demonstrate that macrophages will rapidly switch from trogocytosis to phagocytosis when membrane tension is increased. Stiffening the cortex of target cells also biases macrophages to phagocytose them, a trend that can be countered by increasing antibody surface density and is captured in a mechanical model of trogocytosis. This work suggests that a distinct molecular pathway for trogocytosis is not required to explain differences in trogocytosis among target cell types and points to a mechanism for target cells to modulate trogocytosis.
]]></description>
<dc:creator>Cornell, C. E.</dc:creator>
<dc:creator>Chorlay, A.</dc:creator>
<dc:creator>Krishnamurthy, D.</dc:creator>
<dc:creator>Martin, N. R.</dc:creator>
<dc:creator>Baldauf, L.</dc:creator>
<dc:creator>Fletcher, D. A.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.02.626490</dc:identifier>
<dc:title><![CDATA[Target cell tension regulates macrophage trogocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.05.626924v1?rss=1">
<title>
<![CDATA[
Convergence of autism proteins at the cilium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.05.626924v1?rss=1</link>
<description><![CDATA[
Hundreds of high-confidence autism genes have been identified, yet the relevant etiological mechanisms remain unclear. Gene ontology analyses have repeatedly identified enrichment of proteins with annotated functions in gene expression regulation and neuronal communication. However, proteins are often pleiotropic and these annotations are inherently incomplete. Our recent autism functional genetics work has suggested that these genes may share a common mechanism at the cilium, a membrane-bound organelle critical for neurogenesis, brain patterning, and neuronal activity-all processes strongly implicated in autism. Moreover, autism commonly co-occurs with conditions that are known to involve ciliary-related pathologies, including congenital heart disease, hydrocephalus, and blindness. However, the role of autism genes at the cilium has not been systematically investigated. Here we demonstrate that autism proteins spanning disparate functional annotations converge in expression, localization, and function at cilia, and that patients with pathogenic variants in these genes have cilia-related co-occurring conditions and biomarkers of disrupted ciliary function. This degree of convergence among genes spanning diverse functional annotations strongly suggests that cilia are relevant to autism, as well as to commonly co-occurring conditions, and that this organelle should be explored further for therapeutic potential.

One-Sentence SummaryAutism genes of diverse functional annotations converge at cilia, deepening our understanding of underlying autism biology and co-occurring conditions.
]]></description>
<dc:creator>Kostyanovskaya, E.</dc:creator>
<dc:creator>Lasser, M. C.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Schmidt, J.</dc:creator>
<dc:creator>Bader, E.</dc:creator>
<dc:creator>Buteo, C.</dc:creator>
<dc:creator>Arbelaez, J.</dc:creator>
<dc:creator>Sindledecker, A. R.</dc:creator>
<dc:creator>McCluskey, K. E.</dc:creator>
<dc:creator>Castillo, O.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Dea, J.</dc:creator>
<dc:creator>Helde, K. A.</dc:creator>
<dc:creator>Graglia, J. M.</dc:creator>
<dc:creator>Brimble, E.</dc:creator>
<dc:creator>Kastner, D. B.</dc:creator>
<dc:creator>Ehrlich, A. T.</dc:creator>
<dc:creator>Willsey, A. J.</dc:creator>
<dc:creator>State, M. W.</dc:creator>
<dc:creator>Willsey, H. R.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.05.626924</dc:identifier>
<dc:title><![CDATA[Convergence of autism proteins at the cilium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.03.626573v1?rss=1">
<title>
<![CDATA[
Distinct differentiation trajectories leave lasting impacts on gene regulation and function of V2a interneurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.03.626573v1?rss=1</link>
<description><![CDATA[
During development, early regionalization segregates lineages and directs diverse cell fates. Sometimes, however, distinct progenitors produce analogous cell types. For example, V2a neurons, are excitatory interneurons that emerge from different anteroposterior progenitors. V2a neurons demonstrate remarkable plasticity after spinal cord injury and improve motor function, showing potential for cell therapy. To examine how lineage origins shape their properties, we differentiated V2a neurons from hPSC-derived progenitors with distinct anteroposterior identities. Single-nucleus multiomic analysis revealed lineage-specific transcription factor motifs and numerous differentially expressed genes related to axon growth and calcium handling. Bypassing lineage patterning via transcription factor-induced differentiation yielded neurons distinct from both developmentally relevant populations and human tissue, emphasizing the need to follow developmental steps to generate authentic cell identities. Using in silico and in vitro loss-of-function analyses, we identified CREB5 and TCF7L2 as regulators specific to posterior identities, underscoring the critical role of lieage origins in determining cell states and functions.
]]></description>
<dc:creator>Elder, N. H.</dc:creator>
<dc:creator>Majd, A.</dc:creator>
<dc:creator>Bulger, E. A.</dc:creator>
<dc:creator>Samuel, R. M.</dc:creator>
<dc:creator>Zholudeva, L. V.</dc:creator>
<dc:creator>McDevitt, T. C.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:date>2024-12-06</dc:date>
<dc:identifier>doi:10.1101/2024.12.03.626573</dc:identifier>
<dc:title><![CDATA[Distinct differentiation trajectories leave lasting impacts on gene regulation and function of V2a interneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.06.627170v1?rss=1">
<title>
<![CDATA[
In vivo detection of HIV-1 antisense transcripts in untreated and ART-treated individuals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.06.627170v1?rss=1</link>
<description><![CDATA[
Natural antisense transcripts are expressed in eukaryotes, prokaryotes, and viruses and can possess regulatory functions at the transcriptional and/or post-transcriptional levels. In vitro studies have shown that HIV-1 antisense transcripts (AST) promote viral latency through epigenetic silencing of the proviral 5' long terminal repeat (LTR). However, expression of HIV-1 AST in vivo have not been convincingly demonstrated. Here, we used single RNA template amplification, detection, and sequencing to demonstrate expression of AST in unstimulated PBMC collected from people with HIV-1 (PWH). We found that AST had high genetic diversity that matched proviruses in cells from blood and lymph nodes. We measured a median of 26 copies of AST per 100 infected cells in PWH on ART and a median of 2 copies per 100 infected cells in PWH not on ART. The expression of HIV-1 AST in vivo is consistent with a potential regulatory role in regulation of HIV-1 expression.
]]></description>
<dc:creator>Capoferri, A. A.</dc:creator>
<dc:creator>Sklutuis, R.</dc:creator>
<dc:creator>Famuyiwa, T. O.</dc:creator>
<dc:creator>Pathak, S.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Rausch, J. W.</dc:creator>
<dc:creator>Luke, B. T.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Mellors, J. W.</dc:creator>
<dc:creator>Coffin, J. M.</dc:creator>
<dc:creator>Groebner, J. L.</dc:creator>
<dc:creator>Romerio, F.</dc:creator>
<dc:creator>Kearney, M. F.</dc:creator>
<dc:date>2024-12-06</dc:date>
<dc:identifier>doi:10.1101/2024.12.06.627170</dc:identifier>
<dc:title><![CDATA[In vivo detection of HIV-1 antisense transcripts in untreated and ART-treated individuals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.03.626612v1?rss=1">
<title>
<![CDATA[
High KIR diversity in Uganda and Botswana children living with HIV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.03.626612v1?rss=1</link>
<description><![CDATA[
Killer-cell immunoglobulin-like receptors (KIRs) are essential components of the innate immune system found on the surfaces of natural killer (NK) cells. The KIRs encoding genes are located on chromosome 19q13.4 and are genetically diverse across populations. KIRs are associated with various disease states including HIV progression, and are linked to transplantation rejection and reproductive success. However, there is limited knowledge on the diversity of KIRs from Uganda and Botswana HIV-infected paediatric cohorts, with high endemic HIV rates. We used next-generation sequencing technologies on 312 (246 Uganda, 66 Botswana) samples to generate KIR allele data and employed customised bioinformatics techniques for allelic, allotype and disease association analysis. We show that these sample sets from Botswana and Uganda have different KIRs of different diversities. In Uganda, we observed 147 vs 111 alleles in the Botswana cohort, which had a more than 1 % frequency. We also found significant deviation towards homozygosity for the KIR3DL2 gene for both rapid (RPs) and long-term non-progressors (LTNPs)in the Ugandan cohort. The frequency of the bw4-80I ligand was also significantly higher among the LTNPs than RPs (8.9 % Vs 2.0%, P-value: 0.032). In the Ugandan cohort, KIR2DS4*001 (OR: 0.671, 95 % CI: 0.481-0.937, FDR adjusted Pc=0.142) and KIR2DS4*006 (OR: 2.519, 95 % CI: 1.085-5.851, FDR adjusted Pc=0.142) were not associated with HIV disease progression after adjustment for multiple testing. Our study results provide additional knowledge of the genetic diversity of KIRs in African populations and provide evidence that will inform future immunogenetics studies concerning human disease susceptibility, evolution and host immune responses.
]]></description>
<dc:creator>John, M.</dc:creator>
<dc:creator>Kyobe, S.</dc:creator>
<dc:creator>Amujal, M.</dc:creator>
<dc:creator>Katagirya, E.</dc:creator>
<dc:creator>Diphoko, T.</dc:creator>
<dc:creator>Sebetso, G.</dc:creator>
<dc:creator>Mwesigwa, S.</dc:creator>
<dc:creator>Mboowa, G.</dc:creator>
<dc:creator>Retshabile, G.</dc:creator>
<dc:creator>Williams, L.</dc:creator>
<dc:creator>Mogomotsi, M.</dc:creator>
<dc:creator>Mlotshwa, B.</dc:creator>
<dc:creator>Jjingo, D.</dc:creator>
<dc:creator>kateete, D. P.</dc:creator>
<dc:creator>Joloba, M.</dc:creator>
<dc:creator>Mardon, G.</dc:creator>
<dc:creator>Hanchard, N.</dc:creator>
<dc:creator>Hollenbach, J. A.</dc:creator>
<dc:date>2024-12-07</dc:date>
<dc:identifier>doi:10.1101/2024.12.03.626612</dc:identifier>
<dc:title><![CDATA[High KIR diversity in Uganda and Botswana children living with HIV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.08.627385v1?rss=1">
<title>
<![CDATA[
STING agonists drive recruitment and intrinsic type I interferon responses in monocytic lineage cells for optimal anti-tumor immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.08.627385v1?rss=1</link>
<description><![CDATA[
The cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway, a sensor of cytosolic DNA, orchestrates the production of pro-inflammatory cytokines, chemokines, and type I interferons (IFN-I), thereby contributing to spontaneous tumor surveillance. Intratumoral delivery of synthetic STING agonists induces IFN-I dependent tumor regression in preclinical cancer models and is being tested clinically. In this study, we investigate the role of monocytic lineage (MCs) cells in response to STING agonist induced IFN-I signaling. We show that CCR2-deficient mice, lacking inflammatory MCs in the periphery, or Lyz2-Cre-IFNAR1fl/fl mice in which IFN-I signaling in monocytes is reduced, exhibit impaired responses to STING agonist therapy of MC38 and/or B16F10 tumors. STING agonist treatment induced CCR5-dependent migration of MCs carrying tumor antigen from the tumor to the lymph nodes. Single-cell RNA sequencing of CD45+ cells from lymph nodes and tumors of mice in which half the hematopoietic cells lack the interferon alpha/beta receptor 1 (IFNAR1) revealed that STING agonist therapy induces intrinsic IFNAR1-dependent acquisition of an inflammatory monocytic cell phenotype distinct from inflammatory cDC and a reduction in macrophages with a pro-tumor TGF{beta}/angiogenesis transcriptome. IL-18 - IL-18R1 interaction was the top predicted interaction between monocytic lineage cells and CD8+T cells or NK cells. Blocking IL-18 reduced IFN-{gamma} production by CD8 T cells in LNs and decreased the therapeutic efficacy of STING agonist treatment in CCR2+/+ but not in CCR2-/- mice. These findings support a pivotal role for IL-18 producing inflammatory monocytic lineage cells in CD8 T cell control of melanoma following STING agonist treatment.
]]></description>
<dc:creator>Girard, M.</dc:creator>
<dc:creator>Yu, T.</dc:creator>
<dc:creator>Batista, N. V.</dc:creator>
<dc:creator>Yeung, K. K.</dc:creator>
<dc:creator>Lamorte, S.</dc:creator>
<dc:creator>Gao, W.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>McGaha, T. L.</dc:creator>
<dc:creator>Watts, T. H.</dc:creator>
<dc:date>2024-12-11</dc:date>
<dc:identifier>doi:10.1101/2024.12.08.627385</dc:identifier>
<dc:title><![CDATA[STING agonists drive recruitment and intrinsic type I interferon responses in monocytic lineage cells for optimal anti-tumor immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.10.627829v1?rss=1">
<title>
<![CDATA[
Downstream branches of receptor tyrosine kinase signaling act interdependently to shape the face 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.10.627829v1?rss=1</link>
<description><![CDATA[
BackgroundPreviously we found that increasing fibroblast growth factor (FGF) signaling in the neural crest cells within the frontonasal process (FNP) of the chicken embryo caused dysmorphology that was correlated with reduced proliferation, disrupted cellular orientation, and lower MAPK activation but no change in PLCy and PI3K activation. This suggests RTK signaling may drive craniofacial morphogenesis through specific downstream effectors that affect cellular activities. In this study we inhibited three downstream branches of RTK signaling to determine their role in regulating cellular activities and how these changes affect morphogenesis of the FNP.

ResultsSmall molecule inhibitors of MEK1/2, PI3K, and PLCy were delivered individually and in tandem to the right FNP of chicken embryos. All treatments caused asymmetric proximodistal truncation on the treated side and a mild expansion on the untreated side compared to DMSO control treated FNPs. Inhibiting each pathway caused similar decreased proliferation and disrupted cellular orientation, but did not affect apoptosis.

ConclusionsSince RTK signaling is a ubiquitous and tightly regulated biochemical system we conclude that the downstream pathways are robust to developmental perturbation through redundant signaling systems.

Bullet pointsInhibiting three downstream effectors of receptor tyrosine kinase (RTK) signaling (MEK1/2, PLCy, and PI3K) in the frontonasal process of chicken embryos caused similar mild truncation of growth. Combining all three inhibitors had a slightly stronger effect on truncation.

Individual inhibitors did not have specific effects on cellular proliferation, apoptosis, or cellular orientation.

The downstream branches of RTK signaling likely have shared interdependent effects on cellular activities that contribute to morphogenesis.
]]></description>
<dc:creator>Hanne, N.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Vidal-Garcia, M.</dc:creator>
<dc:creator>Allen, C.</dc:creator>
<dc:creator>Shakir, M. B.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Hallgrimsson, B.</dc:creator>
<dc:creator>Marcucio, R.</dc:creator>
<dc:date>2024-12-11</dc:date>
<dc:identifier>doi:10.1101/2024.12.10.627829</dc:identifier>
<dc:title><![CDATA[Downstream branches of receptor tyrosine kinase signaling act interdependently to shape the face]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.08.627394v1?rss=1">
<title>
<![CDATA[
Enhancer heterogeneity in acute lymphoblastic leukemia drives differential gene expression between patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.08.627394v1?rss=1</link>
<description><![CDATA[
Genetic alterations alone cannot account for the diverse phenotypes of cancer cells. Even cancers with the same driver mutation show significant transcriptional heterogeneity and varied responses to therapy. However, the mechanisms underpinning this heterogeneity remain under-explored. Here, we find that novel enhancer usage is a common feature in acute lymphoblastic leukemia (ALL). In particular, KMT2A::AFF1 ALL, an aggressive leukemia with a poor prognosis and a low mutational burden, exhibits substantial transcriptional heterogeneity between individuals. Using single cell multiome analysis and extensive chromatin profiling, we reveal that much transcriptional heterogeneity in KMT2A::AFF1 ALL is driven by novel enhancer usage. Using high resolution Micro-Capture-C in primary patient samples, we also identify patient-specific enhancer activity at key oncogenes such as MEIS1 and RUNX2, driving high levels of expression of both oncogenes in a patient-specific manner. Overall, our data show that enhancer heterogeneity is highly prevalent in KMT2A::AFF1 ALL and may also be a mechanism that drives transcriptional heterogeneity in cancer more generally.

Key PointsO_LILeukemia patients with the same driver mutations often display gene expression differences
C_LIO_LIUsing chromatin profiling and high resolution 3C methods we show that enhancer heterogeneity drives gene expression differences
C_LI
]]></description>
<dc:creator>Smith, A. L.</dc:creator>
<dc:creator>Denny, N.</dc:creator>
<dc:creator>Chahrour, C.</dc:creator>
<dc:creator>Sharp, K.</dc:creator>
<dc:creator>Elliott, N.</dc:creator>
<dc:creator>Harman, J.</dc:creator>
<dc:creator>Jackson, T.</dc:creator>
<dc:creator>Geng, H.</dc:creator>
<dc:creator>Smith, O.</dc:creator>
<dc:creator>Bond, J.</dc:creator>
<dc:creator>Roberts, I.</dc:creator>
<dc:creator>Stam, R. W.</dc:creator>
<dc:creator>Crump, N. T.</dc:creator>
<dc:creator>Davies, J.</dc:creator>
<dc:creator>Roy, A.</dc:creator>
<dc:creator>Milne, T. A.</dc:creator>
<dc:date>2024-12-12</dc:date>
<dc:identifier>doi:10.1101/2024.12.08.627394</dc:identifier>
<dc:title><![CDATA[Enhancer heterogeneity in acute lymphoblastic leukemia drives differential gene expression between patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.10.627859v1?rss=1">
<title>
<![CDATA[
Recognizing amino acid sidechains in a medium resolution cryo-electron density map 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.10.627859v1?rss=1</link>
<description><![CDATA[
Building an accurate atomic structure model of a protein into a cryo-electron microscopy (cryo-EM) map at worse than 3 [A] resolution is difficult. To facilitate this task, we devised a method for assigning the amino acid residue sequence to the backbone fragments traced in an input cryo-EM map (EMSequenceFinder). EMSequenceFinder relies on a Bayesian scoring function for ranking 20 standard amino acid residue types at a given backbone position, based on the fit to a density map, map resolution, and secondary structure propensity. The fit to a density is quantified by a convolutional neural network that was trained on [~]5.56 million amino acid residue densities extracted from cryo-EM maps at 3-10 [A] resolution and corresponding atomic structure models deposited in the Electron Microscopy Data Bank (EMDB). We benchmarked EMSequenceFinder by predicting the sequences of 58,044 distinct L-helix and {beta}-strand fragments, given the fragment backbone coordinates fitted in their density maps. EMSequenceFinder identifies the correct sequence as the best-scoring sequence in 77.8% of these cases. We also assessed EMSequenceFinder on separate datasets of cryo-EM maps at resolutions from 4 to 6 L. The accuracy of EMSequenceFinder (63.5%) was better than that of three tested state-of-the-art methods, including findMysequence (45%), ModelAngelo (27%), and sequence_from_map in Phenix (12.9%). We further illustrate EMSequenceFinder by threading the SARS-CoV-2 NSP2 sequence into eight cryo-EM maps at resolutions from 3.7 to 7.0 [A]. EMSequenceFinder is implemented in our open-source Integrative Modeling Platform (IMP) program. Thus, it is expected to be helpful for integrative structure modeling based on a cryo-EM map and other information, such as models of protein complex components and chemical crosslinks between them. EMSequenceFinder is available as part of our open source IMP distribution at https://integrativemodeling.org/.
]]></description>
<dc:creator>Mondal, D.</dc:creator>
<dc:creator>Kumar, V.</dc:creator>
<dc:creator>Satler, T.</dc:creator>
<dc:creator>Ramachandran, R.</dc:creator>
<dc:creator>Saltzberg, D.</dc:creator>
<dc:creator>Chemmama, I.</dc:creator>
<dc:creator>Pilla, K. B.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Webb, B. M.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Verba, K. A.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:date>2024-12-12</dc:date>
<dc:identifier>doi:10.1101/2024.12.10.627859</dc:identifier>
<dc:title><![CDATA[Recognizing amino acid sidechains in a medium resolution cryo-electron density map]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.11.627791v1?rss=1">
<title>
<![CDATA[
spotPCR: A Rapid and Efficient Approach for Indexing Individual Template Molecules using Unique Molecular Identifiers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.11.627791v1?rss=1</link>
<description><![CDATA[
Low-frequency mutations provide valuable insights in various fields, including drug resistance identification, cancer and infectious disease research. One promising strategy to enhance the sensitivity and specificity of mutation detection is the incorporation of unique molecular identifiers (UMIs) during polymerase chain reaction (PCR) amplification and before deep sequencing. However, conventional methods for UMI incorporation oRen necessitate multiple labor-intensive steps. spotPCR (Specific Primer Limited Unique Molecular Identifier Tagging PCR) overcomes these challenges, streamlining the UMI tagging process.
]]></description>
<dc:creator>Limberis, J. D.</dc:creator>
<dc:creator>Nagel, R. J.</dc:creator>
<dc:creator>Chakravorty, S.</dc:creator>
<dc:creator>Block, D.</dc:creator>
<dc:creator>Dewell, S.</dc:creator>
<dc:creator>Nalyvayko, A.</dc:creator>
<dc:creator>Metcalfe, J. Z.</dc:creator>
<dc:date>2024-12-12</dc:date>
<dc:identifier>doi:10.1101/2024.12.11.627791</dc:identifier>
<dc:title><![CDATA[spotPCR: A Rapid and Efficient Approach for Indexing Individual Template Molecules using Unique Molecular Identifiers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.13.628212v1?rss=1">
<title>
<![CDATA[
Leucine Aminopeptidase LyLAP enables lysosomal degradation of membrane proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.13.628212v1?rss=1</link>
<description><![CDATA[
Proteolysis of hydrophobic helices is required for complete breakdown of every transmembrane protein trafficked to the lysosome and sustains high rates of endocytosis. However, the lysosomal mechanisms for degrading hydrophobic domains remain unknown. Combining lysosomal proteomics with functional genomic data mining, we identify Lysosomal Leucine Aminopeptidase (LyLAP; formerly Phospholipase B Domain-Containing 1) as the hydrolase most tightly associated with elevated endocytic activity. Untargeted metabolomics and biochemical reconstitution demonstrate that LyLAP is not a phospholipase, but a processive monoaminopeptidase with strong preference for N-terminal leucine - an activity necessary and sufficient for breakdown of hydrophobic transmembrane domains. LyLAP is upregulated in pancreatic ductal adenocarcinoma (PDA), which relies on macropinocytosis for nutrient uptake, and its ablation led to buildup of undigested hydrophobic peptides, which compromised lysosomal membrane integrity and inhibited PDA cell growth. Thus, LyLAP enables lysosomal degradation of membrane proteins, and may represent a vulnerability in highly endocytic cancer cells.

One sentence summaryLyLAP degrades transmembrane proteins to sustain high endocytosis and lysosomal membrane stability in pancreatic cancer.
]]></description>
<dc:creator>Jain, A.</dc:creator>
<dc:creator>Heremans, I.</dc:creator>
<dc:creator>Rademaker, G.</dc:creator>
<dc:creator>Detomasi, T.</dc:creator>
<dc:creator>Rohweder, P.</dc:creator>
<dc:creator>Anderson, D.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Hernandez, G.</dc:creator>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>von Linde, T.</dc:creator>
<dc:creator>Lange, M.</dc:creator>
<dc:creator>Spacci, M.</dc:creator>
<dc:creator>Citron, Y.</dc:creator>
<dc:creator>Olzmann, J.</dc:creator>
<dc:creator>Dawson, D.</dc:creator>
<dc:creator>Craik, C.</dc:creator>
<dc:creator>Bommer, G.</dc:creator>
<dc:creator>Perera, R. M.</dc:creator>
<dc:creator>Zoncu, R.</dc:creator>
<dc:date>2024-12-14</dc:date>
<dc:identifier>doi:10.1101/2024.12.13.628212</dc:identifier>
<dc:title><![CDATA[Leucine Aminopeptidase LyLAP enables lysosomal degradation of membrane proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.13.628239v1?rss=1">
<title>
<![CDATA[
Multiplexed mosaic tumor models reveal natural phenotypic variations in drug response within and between populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.13.628239v1?rss=1</link>
<description><![CDATA[
Many agents that show promise in preclinical cancer models lack efficacy in patients due to patient heterogeneity that is not captured in traditional assays. To address this problem, we have developed GENEVA, a platform that measures the molecular and phenotypic consequences of drug perturbations within diverse populations of cancer cells at single-cell resolution, both in vitro and in vivo. Here, we apply GENEVA to study the KRAS G12C inhibitors, recapitulating known properties of these drugs and uncovering a previously unknown role for mitochondrial activation in cell death induced by KRAS inhibition. We demonstrate that this finding can be leveraged for the development of combination therapies with greater efficacy. Finally, we show that the application of GENEVA with in vivo mouse models revealed epithelial to mesenchymal transition (EMT) as a key mechanism for resistance to KRAS G12C inhibition.
]]></description>
<dc:creator>Yu, J. X.</dc:creator>
<dc:creator>Suh, J. M.</dc:creator>
<dc:creator>Popova, K. D.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Joshi, T.</dc:creator>
<dc:creator>Culbertson, B.</dc:creator>
<dc:creator>Spinelli, J. B.</dc:creator>
<dc:creator>Subramanyam, V.</dc:creator>
<dc:creator>Lou, K.</dc:creator>
<dc:creator>Shokat, K. M.</dc:creator>
<dc:creator>Weissman, J.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2024-12-16</dc:date>
<dc:identifier>doi:10.1101/2024.12.13.628239</dc:identifier>
<dc:title><![CDATA[Multiplexed mosaic tumor models reveal natural phenotypic variations in drug response within and between populations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.15.628586v1?rss=1">
<title>
<![CDATA[
Diverse and Location-Specific Roles of PlexinA2, PlexinA4, and NCAM in Developing Hippocampal Mossy Fibers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.15.628586v1?rss=1</link>
<description><![CDATA[
Mossy fibers (MFs) originate from dentate granule cells and innervate area CA3 of the hippocampus. Upon entry of CA3, MFs partition into two prominent axon bundles, the suprapyramidal tract (SPT) and infrapyramidal tract (IPT) and form lamina specific synaptic contacts in the stratum lucidum (SL) and stratum oriens (SO), respectively. Here we employed new mouse lines to dissect the function of Sema6A and its receptors, PlexinA2 (PlxnA2) and PlxnA4, in developing MFs. In Sema6a-/- mice, MF partitioning into SPT and IPT bundles is incomplete and IPT axons in the SO are overextended, while the SPT correctly innervates the SL. Loss of neuronal Sema6a results in defective MF patterning and we show that this involves Sema6A reverse signaling. Plxna4 controls MF partitioning, SPT axon bundling and laminar targeting to the SL, as well as IPT length. Many of these defects are replicated in mice deficient for PlxnA4 GAP catalytic activity, underscoring the importance of this GAP domain. MFs are tightly fasciculated in Plxna2-/- mice and fail to separate into SPT and IPT bundles, and defects are significantly reduced in PlxnA2 GAP mutants, highlighting the involvement of GAP-independent signaling events. To further explore the molecular basis of aberrant axon fasciculation, we employed anti-PlxnA2 dependent proximity biotinylation and identified several PlxnA2-associated Ig-CAM family members. We observed a genetic interaction between Plxna2 and Ncam1, but not Plxna4 and Ncam1, for SPT and IPT formation and positioning in CA3. Together, our studies provide insights into the multifaceted and overlapping, yet distinct, functions of PlxnA family members in orchestrating specific guidance decisions in developing MFs.
]]></description>
<dc:creator>Zhao, X.-F.</dc:creator>
<dc:creator>Kohen, R.</dc:creator>
<dc:creator>van Battum, E. Y.</dc:creator>
<dc:creator>Zeng, Y.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Johnson, C. N.</dc:creator>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Lim, B. C.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>Rasband, J. M.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Pasterkamp, R. J.</dc:creator>
<dc:creator>Rasband, M. N.</dc:creator>
<dc:creator>Giger, R. J.</dc:creator>
<dc:date>2024-12-16</dc:date>
<dc:identifier>doi:10.1101/2024.12.15.628586</dc:identifier>
<dc:title><![CDATA[Diverse and Location-Specific Roles of PlexinA2, PlexinA4, and NCAM in Developing Hippocampal Mossy Fibers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.16.628616v1?rss=1">
<title>
<![CDATA[
Tax1bp1 enhances bacterial virulence and promotes inflammatory responses during Mycobacterium tuberculosis infection of alveolar macrophages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.16.628616v1?rss=1</link>
<description><![CDATA[
Crosstalk between autophagy, host cell death, and inflammatory host responses to bacterial pathogens enables effective innate immune responses that limit bacterial growth while minimizing coincidental host damage. Mycobacterium tuberculosis (Mtb) thwarts innate immune defense mechanisms in alveolar macrophages (AMs) during the initial stages of infection and in recruited bone marrow-derived cells during later stages of infection. However, how protective inflammatory responses are achieved during Mtb infection and the variation of the response in different macrophage subtypes remain obscure. Here, we show that the autophagy receptor Tax1bp1 plays a critical role in enhancing inflammatory cytokine production and increasing the susceptibility of mice to Mtb infection. Surprisingly, although Tax1bp1 restricts Mtb growth during infection of bone marrow-derived macrophages (BMDMs) (Budzik et al. 2020) and terminates cytokine production in response to cytokine stimulation or viral infection, Tax1bp1 instead promotes Mtb growth in AMs, neutrophils, and a subset of recruited monocyte-derived cells from the bone marrow. Tax1bp1 also leads to increases in bacterial growth and inflammatory responses during infection of mice with Listeria monocytogenes, an intracellular pathogen that is not effectively targeted to canonical autophagy. In Mtb-infected AMs but not BMDMs, Tax1bp1 enhances necrotic-like cell death early after infection, reprogramming the mode of host cell death to favor Mtb replication in AMs. Tax1bp1s impact on host cell death is a mechanism that explains Tax1bp1s cell type-specific role in the control of Mtb growth. Similar to Tax1bp1-deficiency in AMs, the expression of phosphosite-deficient Tax1bp1 restricts Mtb growth. Together, these results show that Tax1bp1 plays a crucial role in linking the regulation of autophagy, cell death, and pro-inflammatory host responses and enhancing susceptibility to bacterial infection.

Author SummaryAlthough macrophages are the first innate immune cells to encounter Mycobacterium tuberculosis during infection, M. tuberculosis has evolved the ability to persist in them. Recent studies highlight that some types of macrophages are more permissive to M. tuberculosis replication and survival than others, but the mechanisms for cell type-specific differences in M. tuberculosis growth are only beginning to be understood. We found that the host factor, Tax1bp1 (Tax-1 binding protein 1), supports M. tuberculosis growth during animal infection and in specific subsets of innate immune cells, including alveolar macrophages while restricting M. tuberculosis in bone marrow-derived macrophages. We also found that Tax1bp1 has a similar phenotype in enhancing the pathogenesis of another intracellular pathogen, Listeria monocytogenes. Compared to bone marrow-derived macrophages, in alveolar macrophages, Tax1bp1 enhances the release of inflammatory mediators and leads to necrotic-like host cell death, which is known to enhance M. tuberculosis growth. Phosphorylation of Tax1bp1 in alveolar macrophages promotes M. tuberculosis growth. Our research highlights that Tax1bp1 is a host target for host-directed therapy against M. tuberculosis and controls host responses to M. tuberculosis in a cell type-specific manner.
]]></description>
<dc:creator>Chin, J.</dc:creator>
<dc:creator>Abeydeera, N.</dc:creator>
<dc:creator>Repasy, T.</dc:creator>
<dc:creator>Rivera-Lugo, R.</dc:creator>
<dc:creator>Mitchell, G.</dc:creator>
<dc:creator>Nguyen, V. Q.</dc:creator>
<dc:creator>Zheng, W.</dc:creator>
<dc:creator>Richards, A.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Ernst, J. D.</dc:creator>
<dc:creator>Cox, J. S.</dc:creator>
<dc:creator>Budzik, J. M.</dc:creator>
<dc:date>2024-12-16</dc:date>
<dc:identifier>doi:10.1101/2024.12.16.628616</dc:identifier>
<dc:title><![CDATA[Tax1bp1 enhances bacterial virulence and promotes inflammatory responses during Mycobacterium tuberculosis infection of alveolar macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.11.628042v1?rss=1">
<title>
<![CDATA[
Patient-derived Induced Pluripotent Stem Cells as a Model to Study Frontotemporal Dementia Pathologies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.11.628042v1?rss=1</link>
<description><![CDATA[
The neurodegenerative disorder Frontotemporal Dementia (FTD) can be caused by a repeat expansion (GGGGCC; G4C2) in C9orf72. The function of wild-type C9orf72 and the mechanism by which the C9orf72-G4C2 mutation causes FTD, however, remain unresolved. Diverse disease models including human brain samples and differentiated neurons from patient-derived induced pluripotent stem cells (iPSCs) identified some hallmarks associated with FTD, but these models have limitations, including biopsies capturing only a static snapshot of dynamic processes and differentiated neurons being labor-intensive, costly, and post-mitotic. We find that patient-derived iPSCs, without being differentiated into neurons, exhibit established FTD hallmarks, including increased lysosome pH, decreased lysosomal cathepsin activity, cytosolic TDP-43 proteinopathy, and increased nuclear TFEB. Moreover, lowering lysosome pH in FTD iPSCs mitigates TDP-43 proteinopathy, suggesting a key role for lysosome dysfunction. RNA-seq reveals dysregulated transcripts in FTD iPSCs affecting calcium signaling, cell death, synaptic function, and neuronal development. We confirm differences in protein expression for some dysregulated genes not previously linked to FTD, including CNTFR (neuronal survival), Annexin A2 (anti-apoptotic), NANOG (neuronal development), and moesin (cytoskeletal dynamics). Our findings underscore the potential of FTD iPSCs as a model for studying FTD cellular pathology and for drug screening to identify therapeutics.

SIGNIFICANCE STATEMENTO_LIUnderstanding the cellular pathology of Frontotemporal Dementia linked to a GGGGCC expansion in the C9orf72 gene remains a challenge.
C_LIO_LIThis study shows that undifferentiated patient-derived iPSCs exhibit hallmark FTD characteristics, including lysosome dysfunction and TDP-43 proteinopathy, and identifies dysregulated genes related to neurodegeneration.
C_LIO_LIThese findings highlight patient-derived iPSCs as a valuable model for studying FTD pathology and for drug screening, potentially guiding future research in therapeutic development.
C_LI
]]></description>
<dc:creator>Barber, D. L.</dc:creator>
<dc:creator>Infante-Tadeo, S.</dc:creator>
<dc:date>2024-12-17</dc:date>
<dc:identifier>doi:10.1101/2024.12.11.628042</dc:identifier>
<dc:title><![CDATA[Patient-derived Induced Pluripotent Stem Cells as a Model to Study Frontotemporal Dementia Pathologies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.16.628706v1?rss=1">
<title>
<![CDATA[
Cell cycle-regulated transcriptional pausing of Drosophila replication-dependent histone genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.16.628706v1?rss=1</link>
<description><![CDATA[
Coordinated expression of replication-dependent (RD) histones genes occurs within the Histone Locus Body (HLB) during S phase, but the molecular steps in transcription that are cell cycle regulated are unknown. We report that Drosophila RNA Pol II promotes HLB formation and is enriched in the HLB outside of S phase, including G1-arrested cells that do not transcribe RD histone genes. In contrast, the transcription elongation factor Spt6 is enriched in HLBs only during S phase. Proliferating cells in the wing and eye primordium express full-length histone mRNAs during S phase but express only short nascent transcripts in cells in G1 or G2 consistent with these transcripts being paused and then terminated. Full-length transcripts are produced when Cyclin E/Cdk2 is activated as cells enter S phase. Thus, activation of transcription elongation by Cyclin E/Cdk2 and not recruitment of RNA pol II to the HLB is the critical step that links histone gene expression to cell cycle progression in Drosophila.
]]></description>
<dc:creator>Kemp, J. P.</dc:creator>
<dc:creator>Geisler, M. S.</dc:creator>
<dc:creator>Hoover, M.</dc:creator>
<dc:creator>Cho, C.-Y.</dc:creator>
<dc:creator>O'Farrell, P. H.</dc:creator>
<dc:creator>Marzluff, W. F.</dc:creator>
<dc:creator>Duronio, R. J.</dc:creator>
<dc:date>2024-12-17</dc:date>
<dc:identifier>doi:10.1101/2024.12.16.628706</dc:identifier>
<dc:title><![CDATA[Cell cycle-regulated transcriptional pausing of Drosophila replication-dependent histone genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.17.628051v1?rss=1">
<title>
<![CDATA[
Small molecule activation of the tumor suppressor kinase LKB1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.17.628051v1?rss=1</link>
<description><![CDATA[
The ability to identify and target oncogenic signals has transformed clinical oncology. Drug development for targeted therapies has historically focused on the inhibition of oncogenic kinases and GTPases. However, many cancer patients do not benefit from targeted approaches because their tumors lack targetable mutations. Therapeutic augmentation of tumor suppressive signaling could be a viable alternative but poses challenges. Specifically, designing compounds capable of stimulating kinase activity is more structurally challenging than inhibitor design, and most kinases lack targetable allosteric pockets. Inactivation of the liver kinase B1 (LKB1) tumor suppressor kinase is associated with poor prognosis and therapeutic resistance. Thus, augmented LKB1 function could be beneficial for cancer patients whose tumors retain intact copies of the gene. LKB1 signals as part of an obligate trimer including the scaffolding protein Mouse protein-25 (MO25) and the pseudokinase (PSK) STE20-related kinase adapter protein (STRAD). As STRAD binds but does not metabolize ATP, it provides the opportunity for a novel activation strategy. We have developed STRAD-binding compounds capable of activating LKB1 and demonstrate the therapeutic benefits of LKB1 activation in a target-dependent manner within cancer cell lines.
]]></description>
<dc:creator>Kretschmer, L. S.</dc:creator>
<dc:creator>Mitchell, D. C.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>He, C.</dc:creator>
<dc:creator>Ding, L.</dc:creator>
<dc:creator>Adler, M.</dc:creator>
<dc:creator>Turnham, R. E.</dc:creator>
<dc:creator>Kellett, T.</dc:creator>
<dc:creator>Keith, A.</dc:creator>
<dc:creator>Hwang, Y. C.</dc:creator>
<dc:creator>Chan, G. K. L.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Ramamoorthi, R.</dc:creator>
<dc:creator>Lively, S.</dc:creator>
<dc:creator>Drakas, R. A.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Qing, T.</dc:creator>
<dc:creator>Verba, K. A.</dc:creator>
<dc:creator>Ostrem, J. M. L.</dc:creator>
<dc:creator>Beresis, R. T.</dc:creator>
<dc:creator>Gordan, J. D.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.17.628051</dc:identifier>
<dc:title><![CDATA[Small molecule activation of the tumor suppressor kinase LKB1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.17.629023v1?rss=1">
<title>
<![CDATA[
HIV-SEQ REVEALS GLOBAL HOST GENE EXPRESSION DIFFERENCES BETWEEN HIV-TRANSCRIBING CELLS FROM VIREMIC AND SUPPRESSED PEOPLE WITH HIV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.17.629023v1?rss=1</link>
<description><![CDATA[
"Active" reservoir cells transcribing HIV can perpetuate chronic inflammation in virally suppressed people with HIV (PWH) and likely contribute to viral rebound after antiretroviral therapy (ART) interruption, so they represent an important target for new therapies. These cells, however, are difficult to study using single-cell RNA-seq (scRNA-seq) due to their low frequency and low levels of HIV transcripts, which are usually not polyadenylated. Here, we developed "HIV-seq" to enable more efficient capture of HIV transcripts - including non-polyadenylated ones - for scRNA-seq analysis of cells from PWH. By spiking in a set of custom-designed capture sequences targeting conserved regions of the HIV genome during scRNA-seq, we increased our ability to find HIV RNA+ cells from PWH by up to 44%. Implementing HIV-seq in conjunction with surface phenotyping by CITE-seq on paired blood specimens from PWH before vs. after ART suppression, we found that HIV RNA+ cells were enriched among T effector memory (Tem) cells during both viremia and ART suppression, but exhibited a cytotoxic signature during viremia only. By contrast, HIV RNA+ cells from the ART-suppressed timepoints exhibited a distinct anti-inflammatory signature involving elevated TGF-{beta} and diminished IFN signaling. Overall, these findings demonstrate that active reservoir cells exhibit transcriptional features distinct from HIV RNA+ cells during viremia, and underscore HIV-seq as a useful tool to better understand the mechanisms by which HIV-transcribing cells can persist during ART.
]]></description>
<dc:creator>Frouard, J.</dc:creator>
<dc:creator>Telwatte, S.</dc:creator>
<dc:creator>Luo, X.</dc:creator>
<dc:creator>Elphick, N.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Arneson, D.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Butte, A. J.</dc:creator>
<dc:creator>Wong, J. K.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Deeks, S.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Roan, N. R.</dc:creator>
<dc:creator>Yukl, S.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.17.629023</dc:identifier>
<dc:title><![CDATA[HIV-SEQ REVEALS GLOBAL HOST GENE EXPRESSION DIFFERENCES BETWEEN HIV-TRANSCRIBING CELLS FROM VIREMIC AND SUPPRESSED PEOPLE WITH HIV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.17.629038v1?rss=1">
<title>
<![CDATA[
Differential roles of NaV1.2 and NaV1.6 in neocortical pyramidal cell excitability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.17.629038v1?rss=1</link>
<description><![CDATA[
Mature neocortical pyramidal cells functionally express two sodium channel (NaV) isoforms: NaV1.2 and NaV1.6. These isoforms are differentially localized to pyramidal cell compartments, and as such are thought to contribute to different aspects of neuronal excitability. But determining their precise roles in pyramidal cell excitability has been hampered by a lack of tools that allow for selective, acute block of each isoform individually. Here, we leveraged aryl sulfonamide-based molecule (ASC) inhibitors of NaV channels that exhibit state-dependent block of both NaV1.2 and NaV1.6, along with knock-in mice with changes in NaV1.2 or NaV1.6 structure that prevents ASC binding. This allowed for acute, potent, and reversible block of individual isoforms that permitted dissection of the unique contributions of NaV1.2 and NaV1.6 in pyramidal cell excitability. Remarkably, block of each isoform had contrasting--and in some situations, opposing--effects on neuronal action potential output, with NaV1.6 block decreasing and NaV1.2 block increasing output. Thus, NaV isoforms have unique roles in regulating different aspects of pyramidal cell excitability, and our work may help guide development of therapeutics designed to temper hyperexcitability through selective NaV isoform blockade.
]]></description>
<dc:creator>Garcia, J. D.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Banks, E.</dc:creator>
<dc:creator>Fenton, T.</dc:creator>
<dc:creator>DeKeyser, J.-M.</dc:creator>
<dc:creator>Abramova, T. V.</dc:creator>
<dc:creator>George, A. L.</dc:creator>
<dc:creator>Ben-Shalom, R.</dc:creator>
<dc:creator>Hackos, D. H.</dc:creator>
<dc:creator>Bender, K. J.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.17.629038</dc:identifier>
<dc:title><![CDATA[Differential roles of NaV1.2 and NaV1.6 in neocortical pyramidal cell excitability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.18.629088v1?rss=1">
<title>
<![CDATA[
The first comorbidity networks in companion dogs in the Dog Aging Project 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.629088v1?rss=1</link>
<description><![CDATA[
Comorbidity and its association with age are of great interest in geroscience. However, there are few model organisms that are well-suited to study comorbidities that will have high relevance to humans. In this light, we turn our attention to the companion dog. The companion dog shares many morbidities with humans. Thus, a better understanding of canine comorbidity relationships could benefit both humans and dogs. We present an analysis of canine comorbidity networks from the Dog Aging Project, a large epidemiological cohort study of companion dogs in the United States. We included owner-reported health conditions that occurred in at least 60 dogs (n=160) and included only dogs that had at least one of those health conditions (n=26,614). We constructed an undirected comorbidity network using a Poisson binomial test, adjusting for age, sex, sterilization status, breed background (i.e., purebred vs. mixed-breed), and weight. The comorbidity network reveals well-documented comorbidities, such as diabetes with cataracts and blindness, and hypertension with chronic kidney disease (CKD). In addition, this network also supports less well-studied comorbidity relationships, such as proteinuria with anemia. A directed comorbidity network accounting for time of reported condition onset suggests that diabetes precedes cataracts, elbow/hip dysplasia before osteoarthritis, and keratoconjunctivitis sicca before corneal ulcer, which are consistent with the canine literature. Analysis of age-stratified networks reveals that global centrality measures increase with age and are the highest in the Senior group compared to the Young Adult and Mature Adult groups. Only the Senior group identified the association between hypertension and CKD. Our results suggest that comorbidity network analysis is a promising method to enhance clinical knowledge and canine healthcare management.

Author SummaryCompanion dogs age alongside humans and suffer many of the same diseases, making them an ideal "real-world" model for human health. Using owner-reported data from 26,614 dogs enrolled in the nationwide Dog Aging Project, we built the first large-scale maps--called comorbidity networks--that show which canine diseases tend to appear together and in what order. The networks correctly highlighted well-known pairings such as diabetes with cataracts and blindness, and hypertension with chronic kidney disease. They also revealed under-appreciated links--for example, protein loss in urine associated with anaemia--suggesting new avenues for veterinary research and care. By adding the reported date of diagnosis, we could infer likely sequences of the diseases: diabetes generally preceded cataracts, hip dysplasia came before osteoarthritis, and dry-eye disease often led to corneal ulcers. When we split the data by life stage, we saw disease webs become denser and more centred on a few key conditions as dogs grew older, echoing patterns seen in people. Together, these findings show that network analysis of large pet-health datasets can guide clinicians, inform breeding and prevention strategies, and ultimately improve the wellbeing of both dogs and humans.
]]></description>
<dc:creator>Fang, A.</dc:creator>
<dc:creator>Kumar, L.</dc:creator>
<dc:creator>Creevy, K. E.</dc:creator>
<dc:creator>Promislow, D. E. L.</dc:creator>
<dc:creator>Ma, J.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629088</dc:identifier>
<dc:title><![CDATA[The first comorbidity networks in companion dogs in the Dog Aging Project]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.18.629283v1?rss=1">
<title>
<![CDATA[
Visualizing Functional Network Connectivity Differences Using an Explainable Machine-learning Method 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.629283v1?rss=1</link>
<description><![CDATA[
Functional network connectivity (FNC) estimated from resting-state functional magnetic resonance imaging showed great information about the neural mechanism in different brain disorders. But previous research has mainly focused on standard statistical learning approaches to find FNC features separating patients from control. Although machine learning approaches provide better models separating controls from patients, it is not straightforward for these approaches to provide intuition on the model and the underlying neural process of each disorder. Explainable machine learning offers a solution to this problem by applying machine learning to understand the neural process behind brain disorders. In this study, we introduce a novel framework leveraging SHapley Additive exPlanations (SHAP) to identify crucial Functional Network Connectivity (FNC) features distinguishing between two distinct population classes. Initially, we validate our approach using synthetic data. Subsequently, applying our framework, we ascertain FNC biomarkers distinguishing between, controls and schizophrenia patients with accuracy of 81.04% as well as middle aged adults and old aged adults with accuracy 71.38%, respectively, employing Random Forest (RF), XGBoost, and CATBoost models. Our analysis underscores the pivotal role of the cognitive control network (CCN), subcortical network (SCN), and somatomotor network (SMN) in discerning individuals with schizophrenia from controls. In addition, our platform found CCN and SCN as the most important networks separating young adults from older.
]]></description>
<dc:creator>Sendi, M. S. E.</dc:creator>
<dc:creator>Itkyal, V. S.</dc:creator>
<dc:creator>Edwards-Swart, S. J.</dc:creator>
<dc:creator>Chun, J. Y.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>Ford, J. M.</dc:creator>
<dc:creator>Preda, A.</dc:creator>
<dc:creator>van Erp, T. G. M.</dc:creator>
<dc:creator>Pearlson, G. D.</dc:creator>
<dc:creator>Turner, J. A.</dc:creator>
<dc:creator>Calhoun, V. D.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629283</dc:identifier>
<dc:title><![CDATA[Visualizing Functional Network Connectivity Differences Using an Explainable Machine-learning Method]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.18.629217v1?rss=1">
<title>
<![CDATA[
Not all cross-modal responses are explained by face movements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.629217v1?rss=1</link>
<description><![CDATA[
The meaning and functions of cross-modal sensory processing in the cortex is at the center of an ongoing debate. While some studies claim that such responses reflect genuine multisensory integration, others argue they are mere artifacts of stimulus-evoked movement or changes in internal state. We examined this issue by measuring face movements and neural activity in awake mouse primary auditory cortex (A1) and primary visual cortex (V1) during visual and auditory stimulation. Visual stimuli rarely evoked face movements, A1 responses to visual input remained robust even in the absence of movement, and optogenetic silencing of V1 reduced A1 visual responses, confirming a sensory origin of these cross-modal responses. These findings directly challenge the argument that cross- modal responses are purely movement-driven and emphasize that rather than assuming all cross-modal effects are artifactual, researchers must rigorously test each case.

HighlightsO_LIUnlike auditory stimuli, visual stimuli rarely evoke face movements
C_LIO_LIFace movements explain sound-evoked firing in VC but not visually evoked firing in AC
C_LIO_LISilencing VC suppresses visually evoked firing in AC
C_LIO_LIThese results confirm a sensory (rather than motor) origin of these responses
C_LI
]]></description>
<dc:creator>Olsen, T. R.</dc:creator>
<dc:creator>Hasenstaub, A. R.</dc:creator>
<dc:date>2024-12-21</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629217</dc:identifier>
<dc:title><![CDATA[Not all cross-modal responses are explained by face movements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.19.629502v1?rss=1">
<title>
<![CDATA[
The cell biology and genome of Stentor pyriformis, a giant cell that embeds symbiotic algae in a microtubule meshwork 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.19.629502v1?rss=1</link>
<description><![CDATA[
Endosymbiotic events in which an endosymbiont is retained within a cell that remains capable of phagocytosis, a situation known as mixotrophy, provide potentially important clues about the evolutionary origins of eukaryotes, particularly regarding the relative evolutionary sequence of phagocytosis and endosymbiosis. Mixotrophy in ciliates is commonplace, but has been investigated at a cellular and molecular level almost entirely in one organism, Paramecium bursaria. Reliance on just one model system makes it difficult to know which cell biological aspects of this system represent general features of ciliate mixotrophy versus accidental features of the specific organism. Here we describe the cell biology and genome of the giant heterotrichous ciliate Stentor pyriformis. We show that this giant unicellular organism contains Chlorella variabilis as its endosymbiont, that the Chlorella can live freely outside the host, that within the host the Chlorella cells are docked near the cell surface, surrounded by microtubule "baskets", and that the photosynthetic efficiency of the Chlorella is reduced inside the Stentor cell compared to when it is free-living outside the host, with photon energy instead being shunted to non-photochemical quenching. Compared to the non-mixotrophic Stentor coeruleus, S. pyriformis has several distinct cellular features that may be related to endosymbiosis: the presence of microtubule baskets, which are absent in S. coeruleus; positive rather than negative phototaxis, which is likely an adaptation to allow the photosynthetic symbionts access to sufficient light; and a lack of pigment in the host cell, which may be an adaptation to tolerate high light levels. Compared to P. bursaria, S. pyriformis has several similar cellular features: in both organisms, the symbiont is a strain of Chlorella variabilis; the Chlorella endosymbiont retains the ability to live freely when separated from the host; and the algal symbionts contained in perialgal vesicles are docked at the cell surface. One potentially informative difference between P. bursaria and S. pyriformis is that S. pyriformis employs a standard genetic code, similar to other Stentor species but different from most other ciliates, including P. bursaria, which use a non-standard code in which one or more stop-codons are respecified to encode amino acids. This difference in genetic code could serve as a barrier to impede gene transfer from symbiont to host in other ciliates, but this would not be a factor in S. pyriformis. A second cell biological difference is that whereas P. bursaria performs phototaxis by a kinetic accumulation mechanism, in which swimming is non-directional but cells slow down in regions of higher light intensity, S. pyriformis performs directed swimming towards the direction of high light intensity. However, as in P. bursaria, phototaxis in S. pyriformis requires the presence of the Chlorella, implying a potential flow of information from the symbiont to direct the orientation and swimming of the host cell. We propose that S. pyriformis will serve as a useful model system for studying the evolution of mixotrophy and endosymbiosis, with unique advantages in terms of size and regenerative ability as well as distinct cellular and genomic features compared with other mixotrophic ciliate models.
]]></description>
<dc:creator>Boudreau, v.</dc:creator>
<dc:creator>Albright, A. R.</dc:creator>
<dc:creator>Gerbich, T.</dc:creator>
<dc:creator>Fadero, T.</dc:creator>
<dc:creator>Yan, V.</dc:creator>
<dc:creator>Larson, B.</dc:creator>
<dc:creator>Lucas-DeMott, A.</dc:creator>
<dc:creator>Yung, J.</dc:creator>
<dc:creator>Moulin, S.</dc:creator>
<dc:creator>Descovich, C. P.</dc:creator>
<dc:creator>Slabodnick, M. M.</dc:creator>
<dc:creator>Burlacot, A.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Niyogi, K. K.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2024-12-21</dc:date>
<dc:identifier>doi:10.1101/2024.12.19.629502</dc:identifier>
<dc:title><![CDATA[The cell biology and genome of Stentor pyriformis, a giant cell that embeds symbiotic algae in a microtubule meshwork]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.20.629759v1?rss=1">
<title>
<![CDATA[
Development and early life stress sensitivity of the rat cortical microstructural similarity network 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.20.629759v1?rss=1</link>
<description><![CDATA[
Understanding how early life experiences shape brain network development is a key challenge in the neuroscience of mental health disorders. To address this, we used magnetic resonance imaging (MRI) similarity network analysis to study the effects of stress in the rat, an important animal model in neuropsychiatry. We measured magnetization transfer ratio (MTR) at each of 53 distinct cortical areas and estimated a cortical similarity network for each individual scan, in two independent experimental datasets. We first characterized normative network development in rats scanned repeatedly between postnatal days 20 (weanling) and 290 (mid-adulthood) (N=47), and then contrasted these findings with a cohort exposed to early life stress in the form of repeated maternal separation (RMS, N=40). The normative rat cortical similarity network exhibited biologically meaningful organization, aligning with prior cytoarchitectonic and tract-tracing data, and displayed complex topological features, including rich club organization. During postnatal and adolescent development, brain regions became more similar, including an early phase of fronto-hippocampal convergence. Early increases in inter-areal similarity were reversed in a later phase of fronto-hippocampal divergence in mid-adulthood. RMS exposure altered inter-areal similarity, especially between frontal and parahippocampal regions, that were also most active developmentally and in aging. Our results reveal how normative cortical network changes in the developing brain are influenced by early life stress. These findings suggest a new translational framework for elucidating how environmental risk factors lead to atypical development of cortical networks.
]]></description>
<dc:creator>Smith, R. L.</dc:creator>
<dc:creator>Sawiak, S. J.</dc:creator>
<dc:creator>Dorfschmidt, L.</dc:creator>
<dc:creator>Dutcher, E. G.</dc:creator>
<dc:creator>Jones, J. A.</dc:creator>
<dc:creator>Hahn, J. D.</dc:creator>
<dc:creator>Sporns, O.</dc:creator>
<dc:creator>Swanson, L. W.</dc:creator>
<dc:creator>Taylor, P. A.</dc:creator>
<dc:creator>Glen, D. R.</dc:creator>
<dc:creator>Dalley, J. W.</dc:creator>
<dc:creator>McMahon, F. J.</dc:creator>
<dc:creator>Raznahan, A.</dc:creator>
<dc:creator>Vertes, P. E.</dc:creator>
<dc:creator>Bullmore, E. T.</dc:creator>
<dc:date>2024-12-21</dc:date>
<dc:identifier>doi:10.1101/2024.12.20.629759</dc:identifier>
<dc:title><![CDATA[Development and early life stress sensitivity of the rat cortical microstructural similarity network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.18.629278v1?rss=1">
<title>
<![CDATA[
Contribution of front-line, standard of care drugs to bactericidal responses, resistance emergence, and cure in murine models of easy- or hard-to-treat tuberculosis disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.629278v1?rss=1</link>
<description><![CDATA[
By assessing the standard of care regimen for tuberculosis (TB) in BALB/c and C3HeB/FeJ mice, we demonstrate that rifampin, with or without pyrazinamide, is essential to effective bactericidal response and resistance suppression. Potency measurements in an in vitro lipid-rich model and rabbit caseum assay recapitulate the significance of rifampin as a sterilizing agent. These outcomes align with clinical performance, thus emphasizing the value of in vitro predictive tools and murine TB models with human-like pathology.
]]></description>
<dc:creator>Peroutka-Bigus, N.</dc:creator>
<dc:creator>Brooks, E. J.</dc:creator>
<dc:creator>Ramey, M. E.</dc:creator>
<dc:creator>D'Erasmo, H.</dc:creator>
<dc:creator>Ernest, J. P.</dc:creator>
<dc:creator>Bauman, A. A.</dc:creator>
<dc:creator>Woolhiser, L. K.</dc:creator>
<dc:creator>Savic, R. M.</dc:creator>
<dc:creator>Lenaerts, A. J.</dc:creator>
<dc:creator>Aldridge, B.</dc:creator>
<dc:creator>Sarathy, J. P.</dc:creator>
<dc:creator>Robertson, G. T.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629278</dc:identifier>
<dc:title><![CDATA[Contribution of front-line, standard of care drugs to bactericidal responses, resistance emergence, and cure in murine models of easy- or hard-to-treat tuberculosis disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.20.629645v1?rss=1">
<title>
<![CDATA[
MRS-Sim: Open-Source Framework for Simulating Realistic In Vivo-like MR Spectroscopy Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.20.629645v1?rss=1</link>
<description><![CDATA[
Realistic, in vivo-like synthetic data is increasingly needed to develop and validate methods in magnetic resonance spectroscopy. MRS-Sim is a powerful, open-source framework for simulating such data while providing known ground truth values. Its modularity enables modeling the complexities of MRS data for various in vivo scenarios. The underlying physical equations include both commonly used spectral components of linear-combination fitting routines and two novel components. The first is a 3D B0 field map simulator that models B0 field inhomogeneities, ranging from slight variations to severe distortions. The second is a novel semi-parametric generator that mimics signals from poorly characterized residual water regions and spectral baseline contributions. This framework can simulate scenarios ranging from raw multi-coil transients to preprocessed, coil-combined multi-average data.

Simulating realistic in vivo-like datasets requires appropriate model parameter ranges and distributions, best determined by analyzing the fitting parameters from existing in vivo data. Therefore, MRS-Sim includes tools for analyzing the ranges and statistical distributions of those parameters from in vivo datasets fitted with Osprey, allowing simulations to be tailored to specific datasets. Additionally, the accompanying repository of supplemental information assists non-expert users with general simulations of MRS data.

The modularity of this framework facilitates easy customization various in vivo scenarios and promotes continued community development. Using a single framework for diverse applications addresses the inconsistencies in current protocols. By simulating in vivo-like data, MRS-Sim supports many MRS tasks, including verifying spectral fitting protocols and conducting reproducibility analyses. Readily available synthetic data also benefits deep learning research, particularly when sufficient in vivo data is unavailable for training. Overall, MRS-Sim will promote reproducibility and make MRS research more accessible to a wider audience.
]]></description>
<dc:creator>LaMaster, J. T.</dc:creator>
<dc:creator>Oeltzschner, G.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.20.629645</dc:identifier>
<dc:title><![CDATA[MRS-Sim: Open-Source Framework for Simulating Realistic In Vivo-like MR Spectroscopy Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.20.629813v1?rss=1">
<title>
<![CDATA[
Haplotype editing with CRISPR/Cas9 as a therapeutic approach for dominant-negative missense mutations in NEFL 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.20.629813v1?rss=1</link>
<description><![CDATA[
Inactivation of disease alleles by allele-specific editing is a promising approach to treat dominant-negative genetic disorders, provided the causative gene is haplo-sufficient. We previously edited a dominant NEFL missense mutation with inactivating frameshifts and rescued disease-relevant phenotypes in induced pluripotent stem cell (iPSC)-derived motor neurons. However, a multitude of different NEFL missense mutations cause disease. Here, we addressed this challenge by targeting common single-nucleotide polymorphisms in cis with NEFL disease mutations for gene excision. We validated this haplotype editing approach for two different missense mutations and demonstrated its therapeutic potential in iPSC-motor neurons. Surprisingly, our analysis revealed that gene inversion, a frequent byproduct of excision editing, failed to reliably disrupt mutant allele expression. We deployed alternative strategies and novel molecular assays to increase therapeutic editing outcomes while maintaining specificity for the mutant allele. Finally, population genetics analysis demonstrated the power of haplotype editing to enable therapeutic development for the greatest number of patients. Our data serve as an important case study for many dominant genetic disorders amenable to this approach.
]]></description>
<dc:creator>Dua, P. H.</dc:creator>
<dc:creator>Simon, B. M.</dc:creator>
<dc:creator>Marley, C. B.</dc:creator>
<dc:creator>Feliciano, C. M.</dc:creator>
<dc:creator>Watry, H. L.</dc:creator>
<dc:creator>Steury, D.</dc:creator>
<dc:creator>Abraham, A.</dc:creator>
<dc:creator>Gilbertson, E. N.</dc:creator>
<dc:creator>Ramey, G. D.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:creator>Judge, L. M.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.20.629813</dc:identifier>
<dc:title><![CDATA[Haplotype editing with CRISPR/Cas9 as a therapeutic approach for dominant-negative missense mutations in NEFL]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.20.629847v1?rss=1">
<title>
<![CDATA[
Protein CREATE enables closed-loop design of de novo synthetic protein binders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.20.629847v1?rss=1</link>
<description><![CDATA[
Proteins have proven to be useful agents in a variety of fields, from serving as potent therapeutics to enabling complex catalysis for chemical manufacture. However, they remain difficult to design and are instead typically selected for using extensive screens or directed evolution. Recent developments in protein large language models have enabled fast generation of diverse protein sequences in unexplored regions of protein space predicted to fold into varied structures, bind relevant targets, and catalyze novel reactions. Nevertheless, we lack methods to characterize these proteins experimentally at scale and update generative models based on those results. We describe Protein CREATE (Computational Redesign via an Experiment-Augmented Training Engine), an integrated computational and experimental pipeline that incorporates an experimental workflow leveraging next generation sequencing and phage display with single-molecule readouts to collect vast amounts of quantitative binding data for updating protein large language models. We use Protein CREATE to generate and assay thousands of designed binders to IL-7 receptor  and insulin receptor with parallel positive and negative selections to identify on-target binders. We discover not only individual novel binders but also features of ligand-receptor binding, including preservation of the IL7R - ligand hydrophobic interface specifically and existence of multiple approaches to contact the insulin receptor. We also demonstrate the importance of structural features, such as the lack of unpaired cysteine residues, toward design fidelity and find computational pre-screening metrics, such as interchain predicted TM scoring (iPTM), while useful, are imperfect predictors as they neither guarantee experimental binding nor rule it out. We use the data collected from Protein CREATE to score designs from the initial generative models. Globally, Protein CREATE will power future closed-loop design-build-test cycles to enable fine-grained design of protein binders.
]]></description>
<dc:creator>Lourenco, A. L.</dc:creator>
<dc:creator>Subramanian, A. M.</dc:creator>
<dc:creator>Spencer, R. K.</dc:creator>
<dc:creator>Miao, J.</dc:creator>
<dc:creator>Anaya, M.</dc:creator>
<dc:creator>Fu, W.</dc:creator>
<dc:creator>Chow, E. D.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.20.629847</dc:identifier>
<dc:title><![CDATA[Protein CREATE enables closed-loop design of de novo synthetic protein binders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.21.629509v1?rss=1">
<title>
<![CDATA[
The biophysical mechanism of mitochondrial pearling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.21.629509v1?rss=1</link>
<description><![CDATA[
Mitochondrial networks exhibit remarkable dynamics that are driven in part by fission and fusion events. However, there are other reorganizations of the network that do not involve fission and fusion. One such exception is the elusive, "beads-on-a-string" morphological transition of mitochondria. During such transitions, the cylindrical tubes of the mitochondrial membrane transiently undergo shape changes to a string of "pearls" connected along thin tubes. These dynamics have been anecdotally observed in many contexts and given disparate explanations. Here we unify these observations by proposing a common underlying mechanism based on the biophysical properties of tubular fluid membranes for which it is known that, under particular regimes of tension and pressure, membranes reach an instability and undergo a shape transition to a string of connected pearls. First, we use high-speed light-sheet microscopy to show that transient, short-lived pearling events occur spontaneously in the mitochondrial network in every cell type we have examined, including primary fibroblasts, T-cells, neurons, and budding yeast. We present evidence that transient mitochondrial pearling occurs during important biological events, particularly during T cell activation, neuronal firing, and replicative senescence. Using our high-temporal resolution data, we identify two distinct categories of spontaneous pearling, i) internal pressure-driven pearling generated by ionic flux, and ii) external tension-driven pearling generated by the cytoskeleton. By applying live-cell STED and FIB-SEM imaging we document the structural reorganization of inner cristae membranes during mitochondrial pearling and the role of the MICOS complex in regulating the frequency of pearling events. We then establish numerous methods for inducing pearling, including the ability to induce these dynamics with single mitochondrion precision. These methods include ionophores, channel activators, osmotic shock, detergents, laser stimulation, membrane intercalating molecules, chemical fixation, and micro-needle force. These disparate inducers establish three main physical causes of pearling, i) ionic flux producing internal osmotic pressure, ii) membrane packing lowering bending elasticity, and iii) external mechanical force increasing membrane tension. Pearling dynamics thereby reveal a fundamental biophysical facet of mitochondrial biology. We suggest that pearling should take its place beside fission and fusion as a key process of mitochondrial dynamics, with implications for physiology, disease, and aging.
]]></description>
<dc:creator>Sturm, G.</dc:creator>
<dc:creator>Hake, K.</dc:creator>
<dc:creator>Lefebvre, A. E. Y. T.</dc:creator>
<dc:creator>Rux, C. J.</dc:creator>
<dc:creator>Ivanova, D.</dc:creator>
<dc:creator>Millett-Sikking, A.</dc:creator>
<dc:creator>Tharp, K. M.</dc:creator>
<dc:creator>Rao, B.</dc:creator>
<dc:creator>Closser, M.</dc:creator>
<dc:creator>Waite, A.</dc:creator>
<dc:creator>Precido-Lopez, M.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:creator>Lu, W.</dc:creator>
<dc:creator>Manley, S.</dc:creator>
<dc:creator>Landoni, J. C.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.21.629509</dc:identifier>
<dc:title><![CDATA[The biophysical mechanism of mitochondrial pearling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.21.629922v1?rss=1">
<title>
<![CDATA[
Collapse of directed functional hierarchy under classical serotonergic psychedelics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.21.629922v1?rss=1</link>
<description><![CDATA[
Despite divergent behavioral and phenomenological profiles, both psychedelic states and reduced states of consciousness have been associated with a flattening of the brains functional hierarchy. To address this apparent paradox, we developed a more specific definition of hierarchy based on the proximity of the brain to thermodynamic equilibrium and then applied it to investigate the changes to the functional hierarchy elicited by three classical serotonergic psychedelics - psilocybin, lysergic acid diethylamide, and dimethyltryptamine. We found that all three psychedelics consistently induced a global reduction in the functional hierarchy. In contrast to the flattening of the functional hierarchy observed during loss of consciousness, psychedelics displaced the brain towards equilibrium while simultaneously increasing the complexity of neural activity, indicating a unique mechanism linked to specific changes in the configuration and differentiation of resting-state networks. This work showcases how metrics based on statistical mechanics can be used for the specific characterization of different global brain states, contributing to the understanding of consciousness as a collective process emerging from complex neural interactions.
]]></description>
<dc:creator>Vohryzek, J.</dc:creator>
<dc:creator>Kringelbach, M. L.</dc:creator>
<dc:creator>Lopez-Sola, E.</dc:creator>
<dc:creator>Garcia Guzman, E.</dc:creator>
<dc:creator>Timmermann, C.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Tagliazucchi, E.</dc:creator>
<dc:creator>Ruffini, G.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Deco, G.</dc:creator>
<dc:creator>Sanz Perl, Y.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.21.629922</dc:identifier>
<dc:title><![CDATA[Collapse of directed functional hierarchy under classical serotonergic psychedelics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.21.629933v1?rss=1">
<title>
<![CDATA[
Spatiotemporal photocatalytic proximity labeling proteomics reveals ligand-activated extracellular and intracellular EGFR neighborhoods 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.21.629933v1?rss=1</link>
<description><![CDATA[
Photo-proximity labeling proteomics (PLP) methods have recently shown that the cell surface receptors can dynamically form lateral interactome networks. Here, we present a paired set of PLP workflows that simultaneously track neighborhood changes for oncogenic epidermal growth factor receptor (EGFR) with temporal resolution, both outside and inside of cells. We achieved this by augmenting the multiscale PLP workflow we call MultiMap, where three photo-probes with different labeling ranges were photo-activated by one photocatalyst, Eosin Y. By anchoring Eosin Y extracellularly and intracellularly on EGFR, we captured hundreds of proteins on both sides of the cell membrane that change in proximity to EGFR upon EGF activation. Neighbors engaged with EGFR within minutes to over an hour, reflecting dynamic interactomes during early, mid- and late-signaling including phosphorylation, internalization, degradation and transcriptional regulation. This rapid "photographic" labeling approach provides snapshots of signaling neighborhoods, revealing their dynamic nature, and potential for drug targeting.
]]></description>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Ngo, W.</dc:creator>
<dc:creator>Chou, Y.-T.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Susa, K. J.</dc:creator>
<dc:creator>Jun, Y.-w.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.21.629933</dc:identifier>
<dc:title><![CDATA[Spatiotemporal photocatalytic proximity labeling proteomics reveals ligand-activated extracellular and intracellular EGFR neighborhoods]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.23.630038v1?rss=1">
<title>
<![CDATA[
Gametocyte production and transmission fitness of African and Asian Plasmodium falciparum isolates with differential susceptibility to artemisinins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.23.630038v1?rss=1</link>
<description><![CDATA[
The emergence of Plasmodium falciparum parasites partially resistant to artemisinins (ART-R) poses a significant threat to recent gains in malaria control. ART-R has been associated with PfKelch13 (K13) mutations, which differ in fitness costs. This study investigates the gametocyte production and transmission fitness of African and Asian P. falciparum isolates with different K13 genotypes across multiple mosquito species. We tested three ART-sensitive (ART-S) isolates (NF54, NF135, NF180) and three ART-R isolates (ARN1G, 3815, PAT-023) for sexual conversion and transmission to Anopheles stephensi, An. gambiae and An. coluzzii. ART-R levels were quantified in vitro using the Ring-stage Survival Assay (RSA), and the transmission-reducing effects of dihydroartemisinin (DHA) on mature gametocytes were assessed. Results showed that ART-S parasite lines consistently produced gametocytes and transmitted effectively in all three mosquito species. ART-R isolates showed variability: ARN1G maintained high transmission levels, whereas 3815 showed limited transmission potential despite higher sporozoite loads in An. coluzzii. The African ART-R isolate PAT-023 demonstrated low gametocyte commitment but was transmitted efficiently in both An. gambiae and An. coluzzii. DHA exposure reduced mosquito infectivity for all isolates, regardless of K13 genotype. These findings, based on a limited number of field isolates, suggest that ART-R parasites remain transmissible across different Anopheles species. However, ART-R does not appear to confer a direct transmission advantage. This study highlights the complexity of ART-R dynamics and underscores the need for further research to inform malaria control strategies in regions where ART-R parasites are circulating.
]]></description>
<dc:creator>Proellochs, N. I.</dc:creator>
<dc:creator>Andolina, C.</dc:creator>
<dc:creator>Ramjith, J.</dc:creator>
<dc:creator>Stoter, R.</dc:creator>
<dc:creator>van Gemert, G.-J.</dc:creator>
<dc:creator>Graumans, W.</dc:creator>
<dc:creator>Campino, S.</dc:creator>
<dc:creator>Vanheer, L. N.</dc:creator>
<dc:creator>Okitwi, M.</dc:creator>
<dc:creator>Tumwebaze, P. K.</dc:creator>
<dc:creator>Conrad, M. D.</dc:creator>
<dc:creator>Clark, T. G.</dc:creator>
<dc:creator>Fidock, D. A.</dc:creator>
<dc:creator>Menard, D.</dc:creator>
<dc:creator>Mok, S.</dc:creator>
<dc:creator>Bousema, T.</dc:creator>
<dc:date>2024-12-23</dc:date>
<dc:identifier>doi:10.1101/2024.12.23.630038</dc:identifier>
<dc:title><![CDATA[Gametocyte production and transmission fitness of African and Asian Plasmodium falciparum isolates with differential susceptibility to artemisinins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.24.630257v1?rss=1">
<title>
<![CDATA[
Inflammatory Monocytes Increase Prior to Detectable HIV-1 Rebound Viremia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.24.630257v1?rss=1</link>
<description><![CDATA[
The persistence of HIV-1 proviruses in latently infected cells allows viremia to resume upon treatment cessation. To characterize the resulting immune response, we compare plasma proteomics and single-cell transcriptomics of peripheral blood mononuclear cells (PBMCs) before, during, and after detectable plasma viremia. We observe unique transcriptional signatures prior to viral rebound including a significant increase in CD16++ monocytes with increased anti-viral gene expression. Inflammatory proteins were identified in plasma after detectable rebound. Identifying early signals of imminent viral rebound after treatment cessation will aid in the development of strategies to prolong time to viral rebound and cure HIV-1.
]]></description>
<dc:creator>Farrell-Sherman, A.</dc:creator>
<dc:creator>de la Force, N.</dc:creator>
<dc:creator>Prator, C.</dc:creator>
<dc:creator>Valieris, R.</dc:creator>
<dc:creator>Azam, W.</dc:creator>
<dc:creator>Silva, I.</dc:creator>
<dc:creator>Deeks, S.</dc:creator>
<dc:creator>Thanh, C.</dc:creator>
<dc:creator>Bosch, R.</dc:creator>
<dc:creator>Henrich, T. J.</dc:creator>
<dc:creator>Cohn, L. B.</dc:creator>
<dc:date>2024-12-25</dc:date>
<dc:identifier>doi:10.1101/2024.12.24.630257</dc:identifier>
<dc:title><![CDATA[Inflammatory Monocytes Increase Prior to Detectable HIV-1 Rebound Viremia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.24.630272v1?rss=1">
<title>
<![CDATA[
Afadin Sorts Different Retinal Neuron Types into Accurate Cellular Layers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.24.630272v1?rss=1</link>
<description><![CDATA[
Neurons use cell-adhesion molecules (CAMs) to interact with other neurons and the extracellular environment: the combination of CAMs specifies migration patterns, neuronal morphologies, and synaptic connections across diverse neuron types. Yet little is known regarding the intracellular signaling cascade mediating the CAM recognitions at the cell surface across different neuron types. In this study, we investigated the neural developmental role of Afadin1-4, a cytosolic adapter protein that connects multiple CAM families to intracellular F-actin. We introduced the conditional Afadin mutant5 to an embryonic retinal Cre, Six3-Cre6-8. We reported that the mutants lead to the scrambled retinal neuron distribution, including Bipolar Cells (BCs), Amacrine Cells (ACs), and retinal ganglion cells (RGCs), across three cellular layers of the retina. This scrambled pattern was first reported here at neuron-type resolution. Importantly, the mutants do not display deficits for BCs, ACs, or RGCs in terms of neural fate specifications or survival. Additionally, the displayed RGC types still maintain synaptic partners with putative AC types, indicating that other molecular determinants instruct synaptic choices independent of Afadin. Lastly, there is a significant decline in visual function and mis-targeting of RGC axons to incorrect zones of the superior colliculus, one of the major retinorecipient areas. Collectively, our study uncovers a unique cellular role of Afadin in sorting retinal neuron types into proper cellular layers as the structural basis for orderly visual processing.
]]></description>
<dc:creator>Lum, M. R.</dc:creator>
<dc:creator>Patel, S.</dc:creator>
<dc:creator>Graham, H. K.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Yi, Y.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Yao, M.</dc:creator>
<dc:creator>La Torre, A.</dc:creator>
<dc:creator>Della Santina, L.</dc:creator>
<dc:creator>Han, Y.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Welsbie, D. S.</dc:creator>
<dc:creator>Duan, X.</dc:creator>
<dc:date>2024-12-25</dc:date>
<dc:identifier>doi:10.1101/2024.12.24.630272</dc:identifier>
<dc:title><![CDATA[Afadin Sorts Different Retinal Neuron Types into Accurate Cellular Layers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.27.630358v1?rss=1">
<title>
<![CDATA[
Integration of Transplanted Interneurons Over a New Period of Ocular Dominance Plasticity in Adult Mouse Visual Cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.27.630358v1?rss=1</link>
<description><![CDATA[
Cortical interneurons play an important role in mediating the juvenile critical period for ocular dominance plasticity in the mouse primary visual cortex. Previously, we showed that transplantation of cortical interneurons derived from the medial ganglionic eminence (MGE) opens a robust period of ocular dominance plasticity 33-35 days after transplantation into neonatal host visual cortex. The plasticity can be induced by transplanting either PV or SST MGE-derived cortical interneurons; it requires transplanted interneurons to express the vesicular GABAergic transporter; and it is manifested by changes to the host visual circuit. Here, we show that transplantation of MGE-derived cortical interneurons into the adult host visual cortex also opens a period of ocular dominance plasticity. The transplanted interneurons must be active to induce plasticity, and the neuronal activity and tuning of visually evoked responses in transplanted and host PV and SST interneurons are modulated by the locomotor state of the host. We also show that changes in activity over the period of plasticity induction are different between PV and SST interneurons but similar between host and transplanted interneurons of each type. The present findings demonstrate that the transplant-induced plasticity generated in adult visual cortex has many features in common with the role of these interneurons during the normal, juvenile critical period.
]]></description>
<dc:creator>Rakela, B.</dc:creator>
<dc:creator>Sun, J.</dc:creator>
<dc:creator>Marchetta, P.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:creator>Hasenstaub, A. R.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:date>2024-12-27</dc:date>
<dc:identifier>doi:10.1101/2024.12.27.630358</dc:identifier>
<dc:title><![CDATA[Integration of Transplanted Interneurons Over a New Period of Ocular Dominance Plasticity in Adult Mouse Visual Cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.27.630563v1?rss=1">
<title>
<![CDATA[
Human precentral gyrus neurons link speech sequences from listening to speaking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.27.630563v1?rss=1</link>
<description><![CDATA[
Speech perception and production are interconnected processes, but the underlying neural mechanisms remain unclear. We investigated this relationship by recording large-scale single-neuron activity in the human brain during a delayed sentence repetition task. Contrary to the traditional view that the precentral gyrus is solely responsible for motor execution, we found that neurons there encoded activity across all task phases of listening, delay, initiation, and speaking. Notably, we discovered "mirror" neurons that activated transiently after hearing and before producing specific speech sounds, and "bridge" neurons that maintained activity between the same speech elements during listening and speaking. Population analysis revealed distinct latent components for each task phase and persistent dynamics for specific sentences. Overall, this study provides novel insights into the neuronal basis of speech processing, emphasizing the intricate interplay between perception, production, and verbal working memory.
]]></description>
<dc:creator>Xu, D.</dc:creator>
<dc:creator>Chung, J. E.</dc:creator>
<dc:creator>Greicius, Q. R.</dc:creator>
<dc:creator>Metzger, S. L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Leonard, M. K.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:date>2024-12-28</dc:date>
<dc:identifier>doi:10.1101/2024.12.27.630563</dc:identifier>
<dc:title><![CDATA[Human precentral gyrus neurons link speech sequences from listening to speaking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.28.630582v1?rss=1">
<title>
<![CDATA[
Convergent representations and spatiotemporal dynamics of speech and language in brain and deep neural networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.28.630582v1?rss=1</link>
<description><![CDATA[
Recent studies have explored the correspondence between single-modality DNN models (speech or text) and specific brain networks for speech and language. The key factors underlying these correlations and their spatiotemporal evolution within the brain language network remain unclear, particularly across different DNN modalities. To address these questions, we analyzed the representation similarity between self-supervised learning (SSL) models for speech (Wav2Vec2) and language (GPT-2), against neural responses to naturalistic speech captured via high-density electrocorticography. Our results indicated high prediction accuracy of both types of SSL models relative to neural activity before and after word onsets. It was the shared components between Wav2Vec2.0 and GPT-2 that explained the majority portion of the SSL-brain similarity. Furthermore, we observed distinct spatiotemporal dynamics: both models showed high encoding accuracy 40 milliseconds before word onset, especially in the mid-superior temporal gyrus (mid-STG), which can be explained by the shared contextual components in the SSL models; the Wav2Vec2.0 also peaked at 200 milliseconds after word onset around the posterior STG, which was mainly attributed to the acoustic-phonetic and static semantic information encoded in the SSL models. These results highlight how contextual and acoustic-phonetic cues encoded in DNNs align with spatiotemporal neural activity patterns, suggesting a significant overlap in how artificial and biological systems process linguistic information.
]]></description>
<dc:creator>Chen, P.</dc:creator>
<dc:creator>Xiang, S.</dc:creator>
<dc:creator>He, L.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:date>2024-12-28</dc:date>
<dc:identifier>doi:10.1101/2024.12.28.630582</dc:identifier>
<dc:title><![CDATA[Convergent representations and spatiotemporal dynamics of speech and language in brain and deep neural networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.30.630730v1?rss=1">
<title>
<![CDATA[
Layilin Regulates Treg Motility and Suppressive Capacity in Skin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.30.630730v1?rss=1</link>
<description><![CDATA[
Regulatory T cells (Tregs) are essential for maintaining immune tolerance in both lymphoid and non-lymphoid tissues. We discovered that layilin, a C-type lectin receptor, is predominantly expressed on Tregs in skin. Layilin was highly expressed on a subset of clonally expanded  effector Tregs in both healthy and psoriatic skin. Layilin expressing Tregs exhibited a transcriptional profile indicative of enhanced adhesion. Deletion of layilin in Tregs in vivo resulted in significantly attenuated skin inflammation. Mechanistically, layilin enhanced Treg adhesion via modulation of LFA-1, resulting in distinct cytoskeletal alterations consistent with enhanced focal adhesion and lamellipodia formation. Taken together, we define layilin as a critical regulator of Treg suppressive capacity through modulating motility and adhesion in a non-lymphoid tissue.
]]></description>
<dc:creator>Gouirand, V.</dc:creator>
<dc:creator>Clancy, S.</dc:creator>
<dc:creator>Macon, C.</dc:creator>
<dc:creator>Valle, J.</dc:creator>
<dc:creator>Pauli, M.</dc:creator>
<dc:creator>Truong, H.-A.</dc:creator>
<dc:creator>Cohen, J.</dc:creator>
<dc:creator>Kinet, M.</dc:creator>
<dc:creator>Lowe, M. M.</dc:creator>
<dc:creator>Lord, S. J.</dc:creator>
<dc:creator>Skruber, K.</dc:creator>
<dc:creator>Harris, H.</dc:creator>
<dc:creator>Kim, E.</dc:creator>
<dc:creator>Neuhaus, I.</dc:creator>
<dc:creator>Reif, K.</dc:creator>
<dc:creator>Zarrin, A. A.</dc:creator>
<dc:creator>Mullins, R. D.</dc:creator>
<dc:creator>Rosenblum, M.</dc:creator>
<dc:date>2024-12-30</dc:date>
<dc:identifier>doi:10.1101/2024.12.30.630730</dc:identifier>
<dc:title><![CDATA[Layilin Regulates Treg Motility and Suppressive Capacity in Skin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.31.630783v1?rss=1">
<title>
<![CDATA[
A generative framework for enhanced cell-type specificity in rationally designed mRNAs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.31.630783v1?rss=1</link>
<description><![CDATA[
mRNA delivery offers new opportunities for disease treatment by directing cells to produce therapeutic proteins. However, designing highly stable mRNAs with programmable cell type-specificity remains a challenge. To address this, we measured the regulatory activity of 60,000 5 and 3 untranslated regions (UTRs) across six cell types and developed PARADE (Prediction And RAtional DEsign of mRNA UTRs), a generative AI framework to engineer untranslated RNA regions with tailored cell type-specific activity. We validated PARADE by testing 15,800 de novo-designed sequences across these cell lines and identified many sequences that demonstrated superior specificity and activity compared to existing RNA therapeutics. mRNAs with PARADE-engineered UTRs also exhibited robust tissue-specific activity in animal models, achieving selective expression in the liver and spleen. We also leveraged PARADE to enhance mRNA stability, significantly increasing protein output and therapeutic durability in vivo. These advancements translated to notable increases in therapeutic efficacy, as PARADE-designed UTRs in oncosuppressor mRNAs, namely PTEN and P16, effectively reduced tumor growth in patient-derived neuroglioma xenograft models and orthotopic mouse models. Collectively, these findings establish PARADE as a versatile platform for designing safer, more precise, and highly stable mRNA therapies.
]]></description>
<dc:creator>Khoroshkin, M. S.</dc:creator>
<dc:creator>Zinkevich, A.</dc:creator>
<dc:creator>Aristova, E.</dc:creator>
<dc:creator>Yousefi, H.</dc:creator>
<dc:creator>Lee, S. B.</dc:creator>
<dc:creator>Mittmann, T.</dc:creator>
<dc:creator>Manegold, K.</dc:creator>
<dc:creator>Penzar, D.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:creator>Kulakovskiy, I. V.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2024-12-31</dc:date>
<dc:identifier>doi:10.1101/2024.12.31.630783</dc:identifier>
<dc:title><![CDATA[A generative framework for enhanced cell-type specificity in rationally designed mRNAs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.31.630834v1?rss=1">
<title>
<![CDATA[
Chronic degenerative failure of salivary glands can be reversed through restoring mitochondrial function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.31.630834v1?rss=1</link>
<description><![CDATA[
Chronic degenerative wounds are often deemed irreparable, directing research efforts to focus predominantly on acute tissue injury regeneration while leaving endogenous repair mechanisms for chronically damaged tissues largely unexplored. In this study, we demonstrate that non-healing, severely degenerated salivary gland tissues can be fundamentally restored through first-line treatment with muscarinic agonists. This approach rescues tissue structure and function, returning it to a homeostatic-like state, and reactivates endogenous regeneration processes to drive new cell expansion that persists for months post-treatment. Furthermore, neuromimetic activation profoundly depletes radiation-induced DNA damage and re-establishes the nerve-acinar relationship, ultimately restoring the tissues physiological capacity to maintain homeostasis, even in the absence of treatment. We show that full recovery of organ function, comparable to uninjured controls, is primarily mediated by the re-differentiation of aberrantly de-differentiated epithelial acinar cells and the restoration of mitochondrial function via a muscarinic-calcium signaling pathway. These findings challenge the prevailing notion that chronic organ degeneration is irreversible and propose a readily testable therapeutic strategy for epithelial restoration with potential applications across a spectrum of chronic injuries.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Tan, L. X.</dc:creator>
<dc:creator>Sun, B.</dc:creator>
<dc:creator>Griffin, N.</dc:creator>
<dc:creator>Niknezhad, S. V.</dc:creator>
<dc:creator>Yu, C.</dc:creator>
<dc:creator>Berthoin, L.</dc:creator>
<dc:creator>Cruz Pacheco, N.</dc:creator>
<dc:creator>Mohabbat, S.</dc:creator>
<dc:creator>Sinada, H.</dc:creator>
<dc:creator>Gaylord, E. A.</dc:creator>
<dc:creator>Chen, Y. T.</dc:creator>
<dc:creator>An, L.</dc:creator>
<dc:creator>Efraim, Y.</dc:creator>
<dc:creator>Bahney, C. S.</dc:creator>
<dc:creator>Lombaert, I. M. A.</dc:creator>
<dc:creator>Knox, S.</dc:creator>
<dc:date>2025-01-02</dc:date>
<dc:identifier>doi:10.1101/2024.12.31.630834</dc:identifier>
<dc:title><![CDATA[Chronic degenerative failure of salivary glands can be reversed through restoring mitochondrial function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.31.630926v1?rss=1">
<title>
<![CDATA[
A20's Linear Ubiquitin Binding Motif Restrains Pathogenic Activation of TH17/22 cells and IL-22 Driven Enteritis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.31.630926v1?rss=1</link>
<description><![CDATA[
A20, encoded by the TNFAIP3 gene, is a protein linked to Crohns disease and celiac disease in humans. We now find that mice expressing point mutations in A20s M1 ubiquitin binding motif (ZF7) spontaneously develop proximate enteritis that requires both luminal microbes and T cells. Cellular and transcriptomic profiling reveal expansion of TH17/22 cells and aberrant expression of IL-17A and IL-22 in intestinal lamina propria of A20ZF7 mice. While deletion of IL-17A from A20ZF7/ZF7 mice exacerbates enteritis, deletion of IL-22 abrogates intestinal epithelial cell hyperproliferation, barrier dysfunction, and alarmin expression. A20ZF7/ZF7 TH17/22 cells autonomously express more ROR{psi}t and IL-22 after differentiation in vitro. ATAC sequencing identified an enhancer region upstream of the Il22 gene in A20ZF7/ZF7 T cells, and this enhancer demonstrated increased activating histone acetylation coupled with exaggerated Il22 transcription. Finally, CRISPR/Cas9-mediated ablation of A20ZF7 in human T cells increases ROR{psi}t expression and IL22 transcription. These studies link A20s M1 ubiquitin binding function with ROR{psi}t expression, epigenetic activation of TH17/22 cells, and IL-22 driven enteritis.
]]></description>
<dc:creator>Bowman, C. J.</dc:creator>
<dc:creator>Stibor, D.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Shimizu, H.</dc:creator>
<dc:creator>Yamashita, E.</dc:creator>
<dc:creator>Lenci, N.</dc:creator>
<dc:creator>Advincula, R.</dc:creator>
<dc:creator>Kim, M. C.</dc:creator>
<dc:creator>Turnbaugh, J. A.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Razani, B.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Malynn, B. A.</dc:creator>
<dc:creator>Ma, A.</dc:creator>
<dc:date>2025-01-02</dc:date>
<dc:identifier>doi:10.1101/2024.12.31.630926</dc:identifier>
<dc:title><![CDATA[A20's Linear Ubiquitin Binding Motif Restrains Pathogenic Activation of TH17/22 cells and IL-22 Driven Enteritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.31.630969v1?rss=1">
<title>
<![CDATA[
Aedes aegypti midgut chymotrypsin has no significant role in blood meal protein digestion or fecundity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.31.630969v1?rss=1</link>
<description><![CDATA[
The Aedes aegypti mosquito is a vector of dengue, Zika, and chikungunya. The mosquitos reliance on blood facilitates the transmission of these viral pathogens to humans. Digestion of blood proteins depends on the biphasic expression of serine proteases, with trypsin-like activity contributing to most of the activity in the midgut. Other proteases found (serine collagenase- and chymotrypsin-like) are thought to contribute to digestion, but their roles are largely understudied. Thus, elucidating the activity and specific roles of all midgut proteases will help understand the complexity of the digestion process and help validate them as potential targets for the development of a new vector control strategy. Herein, we focused on characterizing the activity profile and role of Ae. aegypti chymotrypsin (AaCHYMO). Knockdown studies resulted in elimination and significant reduction of chymotrypsin-like activity in blood fed midgut extracts, while in vitro fluorescent and blood protein digestion assays revealed important substrate specificity differences. Interestingly, knockdown of AaCHYMO did not impact fecundity, indicating the presence of an intricate network of proteases working collectively to degrade blood proteins. Further, knockdown of the ecdysone receptor (EcR) led to a decrease in overall AaCHYMO expression and activity in the mosquito, which may play an important regulatory role.
]]></description>
<dc:creator>Rascon, A. A.</dc:creator>
<dc:creator>Ramirez, A. G.</dc:creator>
<dc:creator>Isoe, J.</dc:creator>
<dc:creator>Serafim, M. S. M.</dc:creator>
<dc:creator>Fong, D.</dc:creator>
<dc:creator>Le, M. A.</dc:creator>
<dc:creator>Nguyen, J. T.</dc:creator>
<dc:creator>Burata, O. E.</dc:creator>
<dc:creator>Lucero, R. M.</dc:creator>
<dc:date>2025-01-02</dc:date>
<dc:identifier>doi:10.1101/2024.12.31.630969</dc:identifier>
<dc:title><![CDATA[Aedes aegypti midgut chymotrypsin has no significant role in blood meal protein digestion or fecundity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.01.631011v1?rss=1">
<title>
<![CDATA[
Perturbing whole-brain models of brain hierarchy: an application for depression following pharmacological treatment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.01.631011v1?rss=1</link>
<description><![CDATA[
Neural representation can extend beyond localised activity to encompass global patterns, where information is distributed across brain networks in a hierarchical manner. Recent research suggests that the hierarchy of causal influences shaping these patterns can serve as a signature of distinct brain states, with implications for neuropsychiatric disorders. Here, we first delve into how whole-brain models, guided by the Thermodynamics of Mind framework, can estimate the brain hierarchy of specific brain states, and how perturbations of such models can study the in-silico transitions to other states represented by static functional connectivity. We then show an application for major depressive disorder, where different brain hierarchical reconfigurations have been found following psilocybin and escitalopram treatments. We build whole-brain models of depressed patients before and after psilocybin and escitalopram interventions, and we carry a dynamic sensitivity analysis to explore the susceptibility of brain states and their drivability to healthier states. We show that susceptibility is on average reduced by escitalopram and increased by psilocybin, and that both treatments succeed in promoting healthier transitions. These results align with the post-treatment window of plasticity opened by serotonergic psychedelics, as well as with the similar clinical efficacy of both drugs observed in clinical trials.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=193 HEIGHT=200 SRC="FIGDIR/small/631011v1_ufig1.gif" ALT="Figure 1">
View larger version (56K):
org.highwire.dtl.DTLVardef@7f1b46org.highwire.dtl.DTLVardef@d7a883org.highwire.dtl.DTLVardef@783e39org.highwire.dtl.DTLVardef@3d334d_HPS_FORMAT_FIGEXP  M_FIG C_FIG We apply whole-brain models of brain hierarchy based on the Thermodynamics of Mind framework to investigate state transitions in depression. Dynamic sensitivity analysis explores how psilocybin and escitalopram affect susceptibility and drivability to healthier states. Results show that psilocybin increases susceptibility, while escitalopram reduces it, with both enabling optimal transitions. This pipeline demonstrates the promise of in-silico approaches to inform neurostimulation protocols, potentially enhancing or complementing antidepressant therapies.
]]></description>
<dc:creator>Socoro Garrigosa, M.</dc:creator>
<dc:creator>Sanz Perl, Y.</dc:creator>
<dc:creator>Kringelbach, M. L.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Vohryzek, J.</dc:creator>
<dc:creator>Deco, G.</dc:creator>
<dc:date>2025-01-02</dc:date>
<dc:identifier>doi:10.1101/2025.01.01.631011</dc:identifier>
<dc:title><![CDATA[Perturbing whole-brain models of brain hierarchy: an application for depression following pharmacological treatment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.02.631038v1?rss=1">
<title>
<![CDATA[
CD38 expression by neonatal human naïve CD4+ T cells shapes their distinct metabolic state and high regulatory T cell potential 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.02.631038v1?rss=1</link>
<description><![CDATA[
Neonatal life is marked by rapid antigen exposure, necessitating establishment of peripheral immune tolerance via conversion of naive CD4+ T cells into regulatory T cells (Tregs). Here, we demonstrate heighted capacity for FOXP3 expression and tolerogenic function among cord blood versus adult blood naive CD4+ T cells and that this is linked to their unique metabolic profile and elevated expression of the NADase, CD38. Early life naive CD4+ T cells demonstrate a metabolic preference for glycolysis, which directly facilitates their differentiation trajectory. We reveal an age-dependent gradient in CD38 levels on naive CD4+ T cells and show that high CD38 expression contributes to both the glycolytic state and tolerogenic potential of neonatal CD4+ T cells, effects that are mediated at least in part via the NAD-dependent deacetylase SIRT1. Thus, the early life window for peripheral tolerance in humans is critically enabled by the immunometabolic state of the naive CD4+ compartment.
]]></description>
<dc:creator>Dwyer, L. R.</dc:creator>
<dc:creator>Clancy, S.</dc:creator>
<dc:creator>Gouirand, V.</dc:creator>
<dc:creator>Chien, C.</dc:creator>
<dc:creator>Rogers, E. E.</dc:creator>
<dc:creator>Oltman, S. P.</dc:creator>
<dc:creator>Jelliffe-Pawlowski, L. L.</dc:creator>
<dc:creator>Lynch, S. V.</dc:creator>
<dc:creator>Wagner, A.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Scharschmidt, T. C.</dc:creator>
<dc:date>2025-01-02</dc:date>
<dc:identifier>doi:10.1101/2025.01.02.631038</dc:identifier>
<dc:title><![CDATA[CD38 expression by neonatal human naïve CD4+ T cells shapes their distinct metabolic state and high regulatory T cell potential]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.02.631109v1?rss=1">
<title>
<![CDATA[
Identifying a next-generation antimalarial trioxolane in a landscape of artemisinin partial resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.02.631109v1?rss=1</link>
<description><![CDATA[
For over two decades, artemisinin-based combination therapy (ACT) has been the standard of care for the treatment of uncomplicated falciparum malaria. However, artemisinin partial resistance (ART-R) is now prevalent in Southeast Asia and has emerged in eastern Africa, threatening ACT efficacy. Mechanistically, ART-R results from an endocytosis defect that limits concentrations of host-derived free heme in the parasite digestive vacuole, allowing early ring-stage parasites to survive exposure to the artemisinin component of ACT. The artemisinin-inspired 1,2,4-trioxolane artefenomel exhibits an extended pharmacokinetic exposure profile that predicts efficacy against ART-R parasites. Unfortunately, the development of artefenomel was halted recently after almost a decade of clinical trials. Herein, we describe the discovery of RLA-4735 and its single-enantiomer form RLA-5764, next-generation antimalarial trioxolanes that exhibit excellent in vitro potency against Plasmodium falciparum and single-exposure efficacy in a murine P. berghei model, thus retaining many of the favorable pharmacokinetic and pharmacodynamic properties of artefenomel while markedly improving solubility and development potential. In P. falciparum samples collected from patients in Uganda in 2019 and 2023, ex vivo ring-stage survival assays revealed the emergence of the ART-R phenotype over this timeframe, and furthermore demonstrated markedly superior activity of artefenomel and RLA-4735 as compared to dihydroartemisinin (the active metabolite of artemisinin components of ACTs) against ART-R parasites. Overall, our findings suggest a role for next-generation trioxolanes in addressing ART-R, and present a potent new, artefenomel-adjacent chemotype with good potential to deliver new development candidates.

Summary SentenceKlope et. al. described the discovery and in vivo characterization of antimalarial endoperoxides effective against artemisinin-resistant parasites as potential development candidates for uncomplicated, blood-stage malaria.
]]></description>
<dc:creator>Klope, M. T.</dc:creator>
<dc:creator>Talukder, P.</dc:creator>
<dc:creator>Blank, B. R.</dc:creator>
<dc:creator>Chelebieva, S.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Fontaine, S. D.</dc:creator>
<dc:creator>Gonciarz, R. L.</dc:creator>
<dc:creator>Jaishankar, P.</dc:creator>
<dc:creator>Lee, G. J.</dc:creator>
<dc:creator>Legac, J.</dc:creator>
<dc:creator>Mathur, V.</dc:creator>
<dc:creator>Narayan, A.</dc:creator>
<dc:creator>Okitwi, M.</dc:creator>
<dc:creator>Orena, S.</dc:creator>
<dc:creator>Settineri, N. S.</dc:creator>
<dc:creator>Tapia, J.</dc:creator>
<dc:creator>Taremwa, Y.</dc:creator>
<dc:creator>Tumwebaze, P. K.</dc:creator>
<dc:creator>Vinod, A.</dc:creator>
<dc:creator>Burrows, J. N.</dc:creator>
<dc:creator>Rosenthal, P. J.</dc:creator>
<dc:creator>Cooper, R. A.</dc:creator>
<dc:creator>Renslo, A. R.</dc:creator>
<dc:date>2025-01-03</dc:date>
<dc:identifier>doi:10.1101/2025.01.02.631109</dc:identifier>
<dc:title><![CDATA[Identifying a next-generation antimalarial trioxolane in a landscape of artemisinin partial resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.02.631118v1?rss=1">
<title>
<![CDATA[
Targeting Osteosarcoma heterogeneity to improve therapeutic response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.02.631118v1?rss=1</link>
<description><![CDATA[
Intra tumor heterogeneity complicates cancer therapy by providing tumors with the ability to alter their phenotypes and become more therapy resistant. Here, we tested the hypothesis that identifying and modulating expression of key state-specific transcription factors could be used as a strategy for driving cells to a more therapy-sensitive state. Recent single-cell studies have explored the inter and intra tumoral heterogeneity of osteosarcoma and identified gene pathways enriched in specific cell states. For example, metastatic tumors are characterized by an expression of genes in the TNF-, PI3K, TGF{beta} and mTOR pathways. We identified similar profiles in osteosarcoma patient-derived xenograft-derived cell lines and potential transcription factor drivers of these states. We then used perturb-seq to downregulate expression of key transcription factors and evaluated the effect of these modulations on single cell RNA profiles and drug responses. Knockdown of NFE2L3 or NR0B1 increased the proportion of cells sensitive to targeted therapy. This approach, which could potentially be applied to other cancers, could be used as a strategy to increase the response to targeted therapies by increasing the proportion of cells in a drug-sensitive state.

HighlightsO_LIDistinct transcriptomic states were identified in osteosarcoma cell lines using single-cell RNA sequencing.
C_LIO_LILineage tracing identified states with differential sensitivity to therapy.
C_LIO_LIUsing perturb-seq, we identified transcription factors that drive cells towards a more sensitive state.
C_LIO_LIThe transcription factor NFE2L3 was identified as targets capable of reprogramming cells to a sensitive state.
C_LI
]]></description>
<dc:creator>Melano, B.</dc:creator>
<dc:creator>Dinh, T.</dc:creator>
<dc:creator>Zirak, B.</dc:creator>
<dc:creator>Karimzadeh, M.</dc:creator>
<dc:creator>Woo, B. J.</dc:creator>
<dc:creator>Choi, B.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Sweet-Cordero, A.</dc:creator>
<dc:date>2025-01-03</dc:date>
<dc:identifier>doi:10.1101/2025.01.02.631118</dc:identifier>
<dc:title><![CDATA[Targeting Osteosarcoma heterogeneity to improve therapeutic response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.02.631150v1?rss=1">
<title>
<![CDATA[
Lactylation fuels nucleotide biosynthesis and facilitates deuterium metabolic imaging of tumor proliferation in H3K27M-mutant gliomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.02.631150v1?rss=1</link>
<description><![CDATA[
Oncogenes hyperactive lactate production, but the mechanisms by which lactate facilitates tumor growth are unclear. Here, we demonstrate that lactate is essential for nucleotide biosynthesis in pediatric diffuse midline gliomas (DMGs). The oncogenic histone H3K27M mutation upregulates phosphoglycerate kinase 1 (PGK1) and drives lactate production from [U-13C]-glucose in DMGs. Lactate activates the nucleoside diphosphate kinase NME1 via lactylation and promotes the synthesis of nucleoside triphosphates essential for tumor proliferation. Importantly, we show that this mechanistic link between glycolysis and nucleotide biosynthesis provides a unique opportunity for deuterium metabolic imaging of DMGs. Spatially mapping 2H-lactate production from [6,6-2H]-glucose allows visualization of the metabolically active tumor lesion and provides an early readout of response to standard-of-care radiation and targeted therapy that precedes extended survival and reflects pharmacodynamic alterations at the tissue level in preclinical DMG models in vivo at clinical field strength (3T). In essence, we have identified an H3K27M-lactate-NME1 axis that promotes DMG proliferation and facilitates non-invasive metabolic imaging of DMGs.

STATEMENT OF SIGNIFICANCEThis study establishes a role for lactate in driving nucleotide biosynthesis in DMGs. Importantly, imaging lactate production from glucose using DMI provides a readout of tumor proliferation and early response to therapy in clinically relevant DMG models. Our studies lay the foundation for precision metabolic imaging of DMG patients.
]]></description>
<dc:creator>Batsios, G.</dc:creator>
<dc:creator>Taglang, C.</dc:creator>
<dc:creator>udutha, s.</dc:creator>
<dc:creator>Gillespie, A. M.</dc:creator>
<dc:creator>Robinson, S. P.</dc:creator>
<dc:creator>Phoenix, T.</dc:creator>
<dc:creator>Mueller, S.</dc:creator>
<dc:creator>Venneti, S.</dc:creator>
<dc:creator>Koschmann, C.</dc:creator>
<dc:creator>Viswanath, P.</dc:creator>
<dc:date>2025-01-03</dc:date>
<dc:identifier>doi:10.1101/2025.01.02.631150</dc:identifier>
<dc:title><![CDATA[Lactylation fuels nucleotide biosynthesis and facilitates deuterium metabolic imaging of tumor proliferation in H3K27M-mutant gliomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.06.631621v1?rss=1">
<title>
<![CDATA[
WormID-Benchmark: Extracting Whole-Brain Neural Dynamics of C. elegans At the Neuron Resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.06.631621v1?rss=1</link>
<description><![CDATA[
The nematode C. elegans is a premier model organism for studying neural circuit function due to its fully mapped connectome and genetically identifiable neurons. Recent advances in 3D light microscopy and fluorescent protein tagging have enabled whole-brain imaging at single-neuron resolution. However, extracting meaningful neural dynamics from these high-resolution recordings requires addressing three fundamental challenges: (i) accurate detection of individual neurons in fluorescence images, (ii) precise identification of neuron classes based on anatomical and colorimetric cues, and (iii) robust tracking of neurons over time in calcium imaging videos. To systematically evaluate these challenges, we introduce WormID-Bench, a large-scale, multi-laboratory dataset comprising 118 worms from five distinct research groups, along with standardized evaluation metrics for detection, identification, and tracking. Our benchmark reveals that existing computational approaches show substantial room for improvement in sensitivity, specificity, and generalization across diverse experimental conditions. By providing an open and reproducible benchmarking framework1, WormID-Bench aims to accelerate the development of high-throughput and scalable computational tools for whole-brain neural dynamics extraction in C. elegans, setting the stage for broader advancements in functional connectomics.
]]></description>
<dc:creator>Adhinarta, J.</dc:creator>
<dc:creator>Dong, J.</dc:creator>
<dc:creator>He, T.</dc:creator>
<dc:creator>Sprague, D.</dc:creator>
<dc:creator>Wan, J.</dc:creator>
<dc:creator>Lee, H. J.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Lu, H.</dc:creator>
<dc:creator>Yemini, E.</dc:creator>
<dc:creator>Kato, S.</dc:creator>
<dc:creator>Varol, E.</dc:creator>
<dc:creator>Wei, D.</dc:creator>
<dc:date>2025-01-07</dc:date>
<dc:identifier>doi:10.1101/2025.01.06.631621</dc:identifier>
<dc:title><![CDATA[WormID-Benchmark: Extracting Whole-Brain Neural Dynamics of C. elegans At the Neuron Resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.08.631999v1?rss=1">
<title>
<![CDATA[
MicroRNA signatures of VO2peak in older adult participants of the Study of Muscle, Mobility and Aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.08.631999v1?rss=1</link>
<description><![CDATA[
BackgroundPeak oxygen consumption during exercise (VO2peak), is a direct measure of cardiorespiratory fitness (CF), a key indicator of physical function and overall health. However, the molecular changes that underpin VO2peak variation are not clear. Our objective is to understand the miRNA signatures that relate to VO2peak variation, which could provide insights to novel mechanisms that contribute to low VO2peak.

MethodsWe used small RNA sequencing to analyze serum samples from 72 participants (70-79 yrs old, 53% female) of the Study of Muscle, Mobility and Aging (SOMMA). We analyzed samples from individuals with low or high VO2peak (N=18/group) as well as samples from 36 randomly selected participants spanning the entire spectrum of VO2peak. We used LIMMA analysis package for regression analysis and to identify differentially expressed miRNAs. We used receiver operating characteristic curve analysis to evaluate the Area Under the Curve (AUC) and sensitivity and specificity rates.

ResultsWe identified 1,055 miRNAs expressed in all serum samples. Expression of 65 miRNAs differed between participants with low and high VO2peak (p < 0.05). After p-value adjustment, expression of 5 miRNAs (miR-1301-3p, -431-5p, -501-5p, -519a-3p, and -18a-3p) remained significantly different (FDR = 0.05). The five miRNAs had AUC ranging from 0.77 to 0.84. The optimal sensitivity and specificity ranged from 70 to 80% and 80 to 90%, respectively. After adjustment for age and sex covariates, 46 miRNAs significantly correlated with VO2peak (p < 0.05) and miR-519a-3p remained significant based on adjusted of p-values.

ConclusionsWe identified a miRNA signature of VO2peak in older individuals that might provide insights to novel mechanisms that drive low VO2peak. Future studies will validate the findings in a larger, longitudinal study cohort.
]]></description>
<dc:creator>Karere, G.</dc:creator>
<dc:creator>Hsu, F.-C.</dc:creator>
<dc:creator>Hepple, R. T.</dc:creator>
<dc:creator>Coen, P. M.</dc:creator>
<dc:creator>Cummings, S.</dc:creator>
<dc:creator>Newman, A.</dc:creator>
<dc:creator>Glynn, N. W.</dc:creator>
<dc:creator>Sparks, L.</dc:creator>
<dc:creator>Lane, N. E.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Wagner, N.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Chan, J.</dc:creator>
<dc:creator>Cox, L. A.</dc:creator>
<dc:creator>Kritchevsky, S.</dc:creator>
<dc:date>2025-01-09</dc:date>
<dc:identifier>doi:10.1101/2025.01.08.631999</dc:identifier>
<dc:title><![CDATA[MicroRNA signatures of VO2peak in older adult participants of the Study of Muscle, Mobility and Aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.08.632037v1?rss=1">
<title>
<![CDATA[
C12ORF57: a novel principal regulator of synaptic AMPA currents and excitatory neuronal homeostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.08.632037v1?rss=1</link>
<description><![CDATA[
ObjectiveExcitatory neuronal homeostasis is crucial for neuronal survival, circuit function, and plasticity. Disruptions in this form of homeostasis are believed to underpin a variety of neuronal conditions including intellectual disability, epilepsy, and autism. However, the underlying genetic and molecular mechanisms maintaining this homeostasis remain poorly understood. Biallelic recurrent loss of function mutations in C12ORF57, an evolutionarily conserved X amino acid novel open reading frame, underlie Temtamy syndrome (TS)--a neurodevelopmental disorder characterized by epilepsy, dysgenesis of the corpus callosum, and severe intellectual disability.

MethodsThrough multiple lines of inquiry, we establish that C12ORF57/GRCC10 plays an unexpected central role in synaptic homeostatic downscaling in response to elevated activity, uncovering a novel mechanism for neuronal excitatory homeostasis. To probe these mechanisms, we developed a new knockout (KO) mouse model of the genes murine ortholog, Grcc10 as well as cellular and in vitro assays.

ResultsGrcc10 KO mice exhibit the characteristic phenotypic features seen in human TS patients, including increased epileptiform activity. Corresponding with the enhanced seizure susceptibility, hippocampal neurons in these mice exhibited significantly increased AMPA receptor expression levels and higher amplitude of miniature excitatory postsynaptic currents (mEPSCs). We further found that GRCC10/C12ORF57 modulates the activity of calcium/calmodulin dependent kinase 4 (CAMK4) and thereby regulates the expression of CREB and ARC.

InterpretationOur study suggests through this novel mechanism, deletion of Grcc10 disrupts the characteristic synaptic AMPA receptor downscaling that accompanies increased activity in glutamatergic neurons.
]]></description>
<dc:creator>Jiang, R.</dc:creator>
<dc:creator>Chouchane, M.</dc:creator>
<dc:creator>Moey, C.</dc:creator>
<dc:creator>Bunt, J.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Allkuraya, F.</dc:creator>
<dc:creator>Alobeid, E.</dc:creator>
<dc:creator>Richards, L.</dc:creator>
<dc:creator>Ullian, E.</dc:creator>
<dc:creator>Sherr, E.</dc:creator>
<dc:date>2025-01-10</dc:date>
<dc:identifier>doi:10.1101/2025.01.08.632037</dc:identifier>
<dc:title><![CDATA[C12ORF57: a novel principal regulator of synaptic AMPA currents and excitatory neuronal homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.09.632276v1?rss=1">
<title>
<![CDATA[
Neural plate pre-patterning enables specification of intermediate neural progenitors in the spinal cord 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.09.632276v1?rss=1</link>
<description><![CDATA[
Dorsal-ventral patterning of neural progenitors in the posterior neural tube, which gives rise to the spinal cord, has served as a model system to understand how extracellular signals organize developing tissues. While previous work has shown that signaling gradients diversify progenitor fates at the dorsal and ventral ends of the tissue, the basis of fate specification in intermediate regions has remained unclear. Here we use zebrafish to investigate the neural plate, which precedes neural tube formation, and show that its pre-patterning by a distinct signaling environment enables intermediate fate specification. Systematic spatial analysis of transcription factor (TF) expression and signaling activity using a reference-based mapping approach shows that the neural plate is partitioned into a striking complexity of TF co-expression states that, in part, correspond to the activity of gastrulation signals such as FGF and Wnt that persist through axis extension. Using in toto analysis of cellular movement combined with fate mapping, we find that dbx1b-expressing intermediate progenitors (p0) originate from a neural-plate specific state characterized by transient co-expression of the TFs pax3a, olig4 and her3. Finally, we show that this state is defined by Wnt signaling in the posterior neural plate and that ectopic Wnt activation within pax3a/olig4+ cells is sufficient to promote dbx1b expression. Our data broadly support a model in which neural progenitor specification occurs through the sequential use of multiple signals to progressively diversify the neural tissue as it develops. This has implications for in vitro differentiation of spinal cord cell types and for understanding signal-based patterning in other developmental contexts.
]]></description>
<dc:creator>Nandagopal, S.</dc:creator>
<dc:creator>Cha, A.</dc:creator>
<dc:creator>Jia, B. Z.</dc:creator>
<dc:creator>Liao, H.</dc:creator>
<dc:creator>Comenho, C.</dc:creator>
<dc:creator>Lahav, G.</dc:creator>
<dc:creator>Wagner, D. E.</dc:creator>
<dc:creator>Tsai, T. Y.-C.</dc:creator>
<dc:creator>Megason, S. G.</dc:creator>
<dc:date>2025-01-10</dc:date>
<dc:identifier>doi:10.1101/2025.01.09.632276</dc:identifier>
<dc:title><![CDATA[Neural plate pre-patterning enables specification of intermediate neural progenitors in the spinal cord]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.09.632202v1?rss=1">
<title>
<![CDATA[
Dose-dependent sensitivity of human 3D chromatin to a heart disease-linked transcription factor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.09.632202v1?rss=1</link>
<description><![CDATA[
Dosage-sensitive transcription factors (TFs) underlie altered gene regulation in human developmental disorders, and cell-type specific gene regulation is linked to the reorganization of 3D chromatin during cellular differentiation. Here, we show dose-dependent regulation of chromatin organization by the congenital heart disease (CHD)- linked, lineage-restricted TF TBX5 in human cardiomyocyte differentiation. Genome organization, including compartments, topologically associated domains, and chromatin loops, are sensitive to reduced TBX5 dosage in a human model of CHD, with variations in response across individual cells. Regions normally bound by TBX5 are especially sensitive, while co-occupancy with CTCF partially protects TBX5-bound TAD boundaries and loop anchors. These results highlight the importance of lineage-restricted TF dosage in cell-type specific 3D chromatin dynamics, suggesting a new mechanism for TF-dependent disease.
]]></description>
<dc:creator>Grant, Z. L.</dc:creator>
<dc:creator>Kuang, S.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Horrillo, A. J.</dc:creator>
<dc:creator>Rao, K. S.</dc:creator>
<dc:creator>Kameswaran, V.</dc:creator>
<dc:creator>Joubran, C.</dc:creator>
<dc:creator>Lau, P. K.</dc:creator>
<dc:creator>Dong, K.</dc:creator>
<dc:creator>Yang, B.</dc:creator>
<dc:creator>Bartosik, W. M.</dc:creator>
<dc:creator>Zemke, N. R.</dc:creator>
<dc:creator>Ren, B.</dc:creator>
<dc:creator>Kathiriya, I. S.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Bruneau, B. G.</dc:creator>
<dc:date>2025-01-12</dc:date>
<dc:identifier>doi:10.1101/2025.01.09.632202</dc:identifier>
<dc:title><![CDATA[Dose-dependent sensitivity of human 3D chromatin to a heart disease-linked transcription factor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.08.632047v1?rss=1">
<title>
<![CDATA[
Viscoelastic HyA Hydrogel Promotes Recovery of Muscle Quality and Vascularization in a Murine Model of Delayed Rotator Cuff Repair 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.08.632047v1?rss=1</link>
<description><![CDATA[
Rotator cuff tears are among the most common musculotendinous injuries with high risk of permanent functional disability. Following surgical repair, sub-optimal patient outcomes are directly correlated with poor muscle quality; namely, injury site fatty infiltration (FI), fibrosis, and muscle atrophy. Muscle resident fibro-adipogenic progenitor cells (FAPs) have been identified as key regulators of post-injury skeletal muscle regeneration and repair by maintaining a promyogenic environment. In this work, human-derived FAPs (hFAPs) were encapsulated into hyaluronic acid (HyA)-based hydrogels functionalized with bsp-RGD(15) cell adhesion peptide, heparin, and a matrix metalloproteinase (MMP)-cleavable crosslinker. Hydrogel-encapsulated hFAPs increased expression of the promyogenic marker UCP1 and production of the anti-inflammatory cytokine IL-10, while downregulating the expression of the fibrotic marker SMA over time. A murine model of unilateral rotator cuff transection, denervation, and delayed repair was treated with the HyA hydrogel or PBS and compared to a contralateral, non-injured control limb. Muscle histology 6 weeks post-repair revealed that the hydrogel reduced fibrosis, FI, and muscle atrophy while supporting vascularization of the injured tissue region. Collectively, these results suggest that the hydrogel alone can promote muscle regeneration in a clinically relevant delayed repair model of rotator cuff tear, which we hypothesize is due to controlled FAP differentiation into promyogenic lineages.
]]></description>
<dc:creator>Pfaff, M. R.</dc:creator>
<dc:creator>Wague, A.</dc:creator>
<dc:creator>Davies, M.</dc:creator>
<dc:creator>Killaars, A. R.</dc:creator>
<dc:creator>Ning, D.</dc:creator>
<dc:creator>Garcia, S.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Nuthalapati, P.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Feeley, B. T.</dc:creator>
<dc:creator>Healy, K. E.</dc:creator>
<dc:date>2025-01-13</dc:date>
<dc:identifier>doi:10.1101/2025.01.08.632047</dc:identifier>
<dc:title><![CDATA[Viscoelastic HyA Hydrogel Promotes Recovery of Muscle Quality and Vascularization in a Murine Model of Delayed Rotator Cuff Repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.09.632255v1?rss=1">
<title>
<![CDATA[
BDNF in Ventrolateral Orbitofrontal Cortex to Dorsolateral Striatum Circuit Moderates Alcohol Consumption and Gates Alcohol Habit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.09.632255v1?rss=1</link>
<description><![CDATA[
BDNF plays a crucial role in shaping the structure and function of neurons. BDNF signaling in the dorsolateral striatum (DLS) is part of an endogenous pathway that protects against the development of alcohol use disorder (AUD). Dysregulation of BDNF levels in the cortex or dysfunction of BDNF/TrkB signaling in the DLS results in the escalation of alcohol drinking and compulsive alcohol use. The major source of BDNF in the striatum is the prefrontal cortex. We identified a small ensemble of BDNF-positive neurons in the ventrolateral orbitofrontal cortex (vlOFC), a region involved in AUD, that extend axonal projections to the DLS. We speculated that BDNF in vlOFC-to-DLS circuit may play a role in limiting alcohol drinking and that heavy alcohol use disrupts this protective pathway. We found that BDNF expression is reduced in the vlOFC of male but not female mice after long-term cycles of binge alcohol drinking and withdrawal. We discovered that overexpression of BDNF in vlOFC-to-DLS but not in vlOFC-to-dorsomedial striatum (DMS) or M2 motor cortex-to-DLS circuit reduces alcohol but not sucrose intake and preference. The DLS plays a major role in habitual behaviors. We hypothesized that BDNF in vlOFC-to-DLS circuitry controls alcohol intake by gating habitual alcohol seeking. We found that BDNF over-expression in vlOFC-to-DLS circuit and systemic administration of BDNF receptor TrkB agonist, LM22A-4, biases habitually trained mice towards goal-directed alcohol seeking. Together, our data suggest that BDNF in a small ensemble of vlOFC-to-DLS neurons gates alcohol intake by attenuating habitual alcohol seeking.
]]></description>
<dc:creator>Gunasekaran, S.</dc:creator>
<dc:creator>Moffat, J. J.</dc:creator>
<dc:creator>Epstein, J. D.</dc:creator>
<dc:creator>Phamluong, K.</dc:creator>
<dc:creator>Ehinger, Y.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:date>2025-01-13</dc:date>
<dc:identifier>doi:10.1101/2025.01.09.632255</dc:identifier>
<dc:title><![CDATA[BDNF in Ventrolateral Orbitofrontal Cortex to Dorsolateral Striatum Circuit Moderates Alcohol Consumption and Gates Alcohol Habit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.11.632543v1?rss=1">
<title>
<![CDATA[
Kynurenine Metabolism is Associated with Antidepressant Response to Selective Serotonin Reuptake Inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.11.632543v1?rss=1</link>
<description><![CDATA[
Alterations in the kynurenine pathway, and in particular the balance of neuroprotective and neurotoxic metabolites, have been implicated in the pathophysiology of Major Depressive Disorder (MDD) and antidepressant treatment response. In this study, we examined the relationship between changes in kynurenine pathway activity (Kynurenine/Tryptophan ratio), focusing on the balance of neuroprotective-to neurotoxic metabolites (Kynurenic Acid/Quinolinic Acid and Kynurenic Acid/3-Hydroxykynurenine ratios), and response to 8 weeks of selective serotonin reuptake inhibitor (SSRI) treatment, including early changes four weeks after SSRI initiation. Additionally, we examined relationships between kynurenine metabolite ratios and three promising biomarkers of depression and antidepressant response: amygdala/hippocampal volume, and glutamate metabolites in the anterior cingulate cortex. Responders showed an increase in the Kynurenic Acid/3-Hydroxykynurenine ratio by week 8 (F(1,46) = 11.92, p = .001) and early increases in the Kynurenine/Tryptophan ratios at week 4 (F(2,58) = 5.224, p = .008), while Non-Responders did not. Pre-treatment Kynurenic Acid/Quinolinic Acid and Kynurenic Acid/3-Hydroxykynurenine ratios were positively associated with right amygdala volume ({beta} = . 247 p = .032 and {beta} = .245 p = .028, respectively). Lastly, in a subset of participants, pre-treatment Kynurenic Acid/3-Hydroxykynurenine ratio showed a positive, small effect size association with glutamate metabolites (Glx) in the anterior cingulate cortex ({beta} = .307 p = .079), which became significant post-treatment with a large effect size ({beta} = .652 p = .021). These results suggest that response to SSRIs may arise from shifting the balance from neurotoxic to neuroprotective kynurenine metabolites.
]]></description>
<dc:creator>RAMPERSAUD, R.</dc:creator>
<dc:creator>Suneson, K.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Lindqvist, D.</dc:creator>
<dc:creator>Ho, T.</dc:creator>
<dc:creator>Myerhoff, D.</dc:creator>
<dc:creator>Reus, V.</dc:creator>
<dc:creator>Irwin, M.</dc:creator>
<dc:creator>Mellon, S.</dc:creator>
<dc:creator>Wolkowitz, O.</dc:creator>
<dc:creator>Brundin, L.</dc:creator>
<dc:date>2025-01-13</dc:date>
<dc:identifier>doi:10.1101/2025.01.11.632543</dc:identifier>
<dc:title><![CDATA[Kynurenine Metabolism is Associated with Antidepressant Response to Selective Serotonin Reuptake Inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.13.632567v1?rss=1">
<title>
<![CDATA[
Predicting Metabolic Dysfunction Associated Steatotic Liver Disease Risk Using Patient-Derived Induced Pluripotent Stem Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.13.632567v1?rss=1</link>
<description><![CDATA[
Background and AimsMetabolic Dysfunction Associated Steatotic Liver Disease (MASLD) is reversible at early stages, making early identification of high-risk individuals clinically valuable. Previously, we demonstrated that patient-derived induced pluripotent stem cells (iPSCs) harboring MASLD DNA risk variants exhibit greater oleate-induced intracellular lipid accumulation than those without these variants. This study aimed to develop an iPSC-based MASLD risk predictor using functional lipid accumulation assessments.

MethodsWe quantified oleate-induced intracellular lipid accumulation in iPSCs derived from three cohorts of diverse ancestry: 1) CIRM cohort (20 biopsy-confirmed MASH cases, 2 biopsy-confirmed MASLD cases, 17 controls), 2) POST cohort (18 MASLD cases, 17 controls), and 3) UCSF cohort (4 biopsy-confirmed MASH cases, 8 controls). Lipid accumulation levels in the CIRM cohort were used to define an iPSC-based MASLD risk score, which was used to predict case/control status in the POST and UCSF cohorts.

ResultsIn all three cohorts, lipid accumulation was higher in MASLD/MASH cases vs. controls (CIRM cases vs. controls 3.32 {+/-} 0.25 vs. 2.70 {+/-} 0.19 -fold change, p=0.06; POST cases vs. controls 3.63 {+/-} 0.33 vs. 2.70 {+/-} 0.31, p=0.05; and UCSF cases vs. controls 4.39{+/-}0.46 vs. 2.03{+/-}0.20, p=0.0002). The iPSC-based MASLD risk score achieved a sensitivity of 44% and specificity of 75% in the POST cohort and 75% and 100%, respectively, in the UCSF cohort. Differences in cohort disease severity and cardiometabolic profiles may explain performance variability.

ConclusionWhile validation in larger cohorts is needed, these findings suggest that oleate-induced intracellular lipid accumulation in subject-derived iPSCs is predictive of MASH development. Additional cellular phenotypes and donor information should be explored to improve predictive accuracy to inform MASLD surveillance and prevention strategies.
]]></description>
<dc:creator>Qin, Y.</dc:creator>
<dc:creator>Chhetri, P.</dc:creator>
<dc:creator>Theusch, E.</dc:creator>
<dc:creator>Lim, G.</dc:creator>
<dc:creator>Teker, S.</dc:creator>
<dc:creator>Kuang, Y.-L.</dc:creator>
<dc:creator>Aziziraftar, S. K.</dc:creator>
<dc:creator>Mehraban, M. H.</dc:creator>
<dc:creator>Munoz-Howell, A.</dc:creator>
<dc:creator>Saxena, V.</dc:creator>
<dc:creator>Le Guillou, D.</dc:creator>
<dc:creator>Mattis, A. N.</dc:creator>
<dc:creator>Maher, J. J.</dc:creator>
<dc:creator>Medina, M. N.</dc:creator>
<dc:date>2025-01-15</dc:date>
<dc:identifier>doi:10.1101/2025.01.13.632567</dc:identifier>
<dc:title><![CDATA[Predicting Metabolic Dysfunction Associated Steatotic Liver Disease Risk Using Patient-Derived Induced Pluripotent Stem Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.14.632786v1?rss=1">
<title>
<![CDATA[
Pseudomonas superinfection drives Pf phage transmission within airway infections in patients with cystic fibrosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.14.632786v1?rss=1</link>
<description><![CDATA[
Pf bacteriophages, lysogenic viruses that infect Pseudomonas aeruginosa (Pa), are implicated in the pathogenesis of chronic Pa infections; phage-infected (Pf+) strains are known to predominate in people with cystic fibrosis (pwCF) who are older and have more severe disease. However, the transmission patterns of Pf underlying the progressive dominance of Pf+ strains are unclear. In particular, it is unknown whether phage transmission commonly occurs horizontally between bacteria within the airway via viral particles or if Pf+ bacteria are mostly acquired via new Pseudomonas infections. Here, we have studied Pa genomic sequences from 3 patient cohorts totaling 663 clinical isolates from 105 pwCF. We identify Pf+ isolates and analyze transmission patterns of Pf within patients between genetically similar groups of bacteria called "clone types". We find that Pf is predominantly passed down vertically within Pa lineages and rarely via horizontal transfer between clone types within the airway. Conversely, we find extensive evidence of Pa superinfection by a new, genetically distinct Pa that is Pf+. Finally, we find that clinical isolates show reduced activity of the type IV pilus and reduced susceptibility to Pf in vitro. These results cast new light on the transmission of virulence-associated phages in the clinical setting.
]]></description>
<dc:creator>Pourtois, J. D.</dc:creator>
<dc:creator>Haddock, N. L.</dc:creator>
<dc:creator>Gupta, A.</dc:creator>
<dc:creator>Khosravi, A.</dc:creator>
<dc:creator>Martinez, H.</dc:creator>
<dc:creator>Schmidt, A. K.</dc:creator>
<dc:creator>Prakash, P. S.</dc:creator>
<dc:creator>Jain, R.</dc:creator>
<dc:creator>Fleming, P.</dc:creator>
<dc:creator>Chang, T. H.</dc:creator>
<dc:creator>Milla, C.</dc:creator>
<dc:creator>Secor, P. R.</dc:creator>
<dc:creator>De Leo, G. A.</dc:creator>
<dc:creator>Bollyky, P. L.</dc:creator>
<dc:creator>Burgener, E. B.</dc:creator>
<dc:date>2025-01-15</dc:date>
<dc:identifier>doi:10.1101/2025.01.14.632786</dc:identifier>
<dc:title><![CDATA[Pseudomonas superinfection drives Pf phage transmission within airway infections in patients with cystic fibrosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.21.634052v1?rss=1">
<title>
<![CDATA[
Defining the cell and molecular origins of the primate ovarian reserve 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.21.634052v1?rss=1</link>
<description><![CDATA[
The primate ovarian reserve is established during late fetal development and consists of quiescent primordial follicles in the ovarian cortex, each composed of granulosa cells surrounding an oocyte in dictate. As late stages of fetal development are not routinely accessible for study with human tissue, we exploited the evolutionary proximity of the rhesus macaque to investigate primate follicle formation. Similar to human prenatal ovaries, the rhesus also develops multiple types of pre-granulosa (PG) cells, with the majority of primordial follicles derived from PG2 with small variable contributions from PG1. We observed that activated medullary follicles recruit fetal theca cells to establish a two-cell system for sex-steroid hormone production prior to birth, providing a cell-based explanation for mini puberty.
]]></description>
<dc:creator>Wamaitha, S. E.</dc:creator>
<dc:creator>Rojas, E. J.</dc:creator>
<dc:creator>Monticolo, F.</dc:creator>
<dc:creator>Hsu, F.-M.</dc:creator>
<dc:creator>Sosa, E.</dc:creator>
<dc:creator>Mackie, A. M.</dc:creator>
<dc:creator>Oyama, K.</dc:creator>
<dc:creator>Custer, M.</dc:creator>
<dc:creator>Murphy, M.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:creator>Shu, J.</dc:creator>
<dc:creator>Hennebold, J. D.</dc:creator>
<dc:creator>Clark, A.</dc:creator>
<dc:date>2025-01-21</dc:date>
<dc:identifier>doi:10.1101/2025.01.21.634052</dc:identifier>
<dc:title><![CDATA[Defining the cell and molecular origins of the primate ovarian reserve]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.20.633978v1?rss=1">
<title>
<![CDATA[
Druggable genome CRISPRi screen in 3D hydrogels reveals regulators of cortactin-driven actin remodeling in invading glioblastoma cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.20.633978v1?rss=1</link>
<description><![CDATA[
To identify new therapeutic targets that limit glioblastoma (GBM) invasion, we applied druggable-genome CRISPR screens to patient-derived GBM cells in micro-dissectible biomimetic 3D hydrogel platforms that permit separation and independent analysis of core vs. invasive fractions. We identified 12 targets whose suppression limited invasion, of which ACP1 (LMW-PTP) and Aurora Kinase B (AURKB) were validated in neurosphere assays. Proximity labeling analysis identified cortactin as an ACP1- AURKB link, as cortactin undergoes serine phosphorylation by AURKB and tyrosine dephosphorylation by ACP1. Suppression of ACP1 or AURKB in culture and in vivo shifted the balance of cortactin phosphorylation in GBM and reduced actin polymerization and actin-cortactin co-localization. Additional biophysical analysis implicated AURKB in GBM cell adhesion and cortical stiffness, and ACP1 in resistance to mechanical stress and shape plasticity needed for 3D migration. These findings reveal a novel targetable axis that balances kinase and phosphatase activities to regulate actin polymerization during GBM invasion.
]]></description>
<dc:creator>Hu, M.</dc:creator>
<dc:creator>Weldy, A.</dc:creator>
<dc:creator>Lovalvo, I.</dc:creator>
<dc:creator>Akins, E.</dc:creator>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Chang, A.</dc:creator>
<dc:creator>Sati, A.</dc:creator>
<dc:creator>Lad, M.</dc:creator>
<dc:creator>Lui, A.</dc:creator>
<dc:creator>Rajidi, A.</dc:creator>
<dc:creator>Kothekar, A.</dc:creator>
<dc:creator>Ding, E. A.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:date>2025-01-24</dc:date>
<dc:identifier>doi:10.1101/2025.01.20.633978</dc:identifier>
<dc:title><![CDATA[Druggable genome CRISPRi screen in 3D hydrogels reveals regulators of cortactin-driven actin remodeling in invading glioblastoma cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.22.634424v1?rss=1">
<title>
<![CDATA[
Causal modeling of gene effects from regulators to programs to traits: integration of genetic associations and Perturb-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.22.634424v1?rss=1</link>
<description><![CDATA[
AbstractGenetic association studies provide a unique tool for identifying causal links from genes to human traits and diseases. However, it is challenging to determine the biological mechanisms underlying most associations, and we lack genome-scale approaches for inferring causal mechanistic pathways from genes to cellular functions to traits. Here we propose new approaches to bridge this gap by combining quantitative estimates of gene-trait relationships from loss-of-function burden tests with gene-regulatory connections inferred from Perturb-seq experiments in relevant cell types. By combining these two forms of data, we aim to build causal graphs in which the directional associations of genes with a trait can be explained by their regulatory effects on biological programs or direct effects on the trait. As a proof-of-concept, we constructed a causal graph of the gene regulatory hierarchy that jointly controls three partially co-regulated blood traits. We propose that perturbation studies in trait-relevant cell types, coupled with gene-level effect sizes for traits, can bridge the gap between genetics and biology.
]]></description>
<dc:creator>Ota, M.</dc:creator>
<dc:creator>Spence, J. P.</dc:creator>
<dc:creator>Zeng, T.</dc:creator>
<dc:creator>Dann, E.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:date>2025-01-24</dc:date>
<dc:identifier>doi:10.1101/2025.01.22.634424</dc:identifier>
<dc:title><![CDATA[Causal modeling of gene effects from regulators to programs to traits: integration of genetic associations and Perturb-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.23.634542v1?rss=1">
<title>
<![CDATA[
Convergent Agonist and Heat Activation of Nociceptor TRPM3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.23.634542v1?rss=1</link>
<description><![CDATA[
Detecting noxious heat is vital for survival, triggering pain responses that protect against harm1,2. The TRPM3 channel is a key nociceptor for sensing noxious heat and a promising therapeutic target for pain treatment and neurological disorders such as epilepsy3-11. Here, we functionally and structurally characterized TRPM3 in response to diverse stimuli: the synthetic superagonist CIM0216 Ref12, the anticonvulsant antagonist primidone13,14, and heat1,10,15. Our findings reveal that TRPM3 is intrinsically dynamic, with its intracellular domain (ICD) sampling both resting and activated states, though strongly favoring the resting state without stimulation. CIM0216 binds to the S1-S4 domain, inducing conformational changes in the ICD and shifting the equilibrium toward activation. Remarkably, heat induces similar ICD rearrangements, revealing a converged activation mechanism driven by chemical compounds and temperature. This mechanism is supported by functional data showing that mutations facilitating the ICD movement markedly increase the sensitivity of TRPM3 to both chemical and thermal signals. These findings establish a critical role of the ICD in temperature sensing in TRPM3, a mechanism likely conserved across the TRPM family. Finally, we show that primidone binds to the same site as CIM0216 but acts as an antagonist. This study provides a framework for understanding the thermal sensing mechanisms of temperature-sensitive ion channels and offers a structural foundation for developing TRPM3-target therapeutics for pain and neurological disorders.
]]></description>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Lu, W.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Jin, F.</dc:creator>
<dc:creator>Park, S. J.</dc:creator>
<dc:creator>Keuning, S. I.</dc:creator>
<dc:creator>Massimino, R. P.</dc:creator>
<dc:creator>Choi, W.</dc:creator>
<dc:creator>Vu, S.</dc:creator>
<dc:date>2025-01-24</dc:date>
<dc:identifier>doi:10.1101/2025.01.23.634542</dc:identifier>
<dc:title><![CDATA[Convergent Agonist and Heat Activation of Nociceptor TRPM3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.23.634631v1?rss=1">
<title>
<![CDATA[
Loss of Fanconi anemia proteins causes a reliance on lysosomal exocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.23.634631v1?rss=1</link>
<description><![CDATA[
Mutations in the FA pathway lead to a rare genetic disease that increases risk of bone marrow failure, acute myeloid leukemia, and solid tumors. FA patients have a 500 to 800-fold increase in head and neck squamous cell carcinoma compared to the general population and the treatment for these malignancies are ineffective and limited due to the deficiency in DNA damage repair. Using unbiased CRISPR-interference screening, we found the loss of FA function renders cells dependent on key exocytosis genes such as SNAP23. Further investigation revealed that loss of FA pathway function induced deficiencies in lysosomal health, dysregulation of autophagy and increased lysosomal exocytosis. The compromised cellular state caused by the loss of FA genes is accompanied with decreased lysosome abundance and increased lysosomal membrane permeabilization in cells. We found these signatures in vitro across multiple cell types and cell lines and in clinically relevant FA patient cancers. Our findings are the first to connect the FA pathway to lysosomal exocytosis and thus expands our understanding of FA as a disease and of induced dependencies in FA mutant cancers.
]]></description>
<dc:creator>Fu, B. X. H. X. H.</dc:creator>
<dc:creator>Xu, A.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Grandis, J. R.</dc:creator>
<dc:creator>Johnson, D. E.</dc:creator>
<dc:creator>Gilbert, L.</dc:creator>
<dc:date>2025-01-24</dc:date>
<dc:identifier>doi:10.1101/2025.01.23.634631</dc:identifier>
<dc:title><![CDATA[Loss of Fanconi anemia proteins causes a reliance on lysosomal exocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.25.634870v1?rss=1">
<title>
<![CDATA[
URAdime - a tool for analyzing primer sequences in sequencing data to identify dimers and super-amplicons. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.25.634870v1?rss=1</link>
<description><![CDATA[
Targeted sequencing of multiplex PCR amplicons is commonplace in research laboratories and clinical diagnostics. There are numerous tools for the a priori optimization of primers and reactions, but no tools to detect specific problematic primers post hoc. We developed URAdime, a tool for analyzing primer sequences in sequencing data to identify dimers and super-amplicons. We show that it successfully detects these unwanted amplicons and accurately attributes their generation to specific primers.
]]></description>
<dc:creator>Limberis, J. D.</dc:creator>
<dc:creator>Metcalfe, J. Z.</dc:creator>
<dc:date>2025-01-28</dc:date>
<dc:identifier>doi:10.1101/2025.01.25.634870</dc:identifier>
<dc:title><![CDATA[URAdime - a tool for analyzing primer sequences in sequencing data to identify dimers and super-amplicons.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.27.635090v1?rss=1">
<title>
<![CDATA[
Nemo-like kinase disrupts nuclear import and drives TDP43 mislocalization in ALS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.27.635090v1?rss=1</link>
<description><![CDATA[
Cytoplasmic TDP43 mislocalization and aggregation are pathological hallmarks of amyotrophic lateral sclerosis (ALS). However, the initial cellular insults that lead to TDP43 mislocalization remain unclear. In this study, we demonstrate that Nemo-like kinase (NLK)--a proline-directed serine/threonine kinase--promotes the mislocalization of TDP43 and other RNA-binding proteins by disrupting nuclear import. NLK levels are selectively elevated in neurons exhibiting TDP43 mislocalization in ALS patient tissues, while genetic reduction of NLK reduces toxicity in human neuron models of ALS. Our findings suggest that NLK is a promising therapeutic target for neurodegenerative diseases.
]]></description>
<dc:creator>Bekier, M. E.</dc:creator>
<dc:creator>Pinarbasi, E.</dc:creator>
<dc:creator>Mesojedec, J. J.</dc:creator>
<dc:creator>Ghaffari, L.</dc:creator>
<dc:creator>de Majo, M.</dc:creator>
<dc:creator>Ullian, E.</dc:creator>
<dc:creator>Koontz, M.</dc:creator>
<dc:creator>Coleman, S.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Tank, E. M.</dc:creator>
<dc:creator>Waksmacki, J.</dc:creator>
<dc:creator>Barmada, S.</dc:creator>
<dc:date>2025-01-28</dc:date>
<dc:identifier>doi:10.1101/2025.01.27.635090</dc:identifier>
<dc:title><![CDATA[Nemo-like kinase disrupts nuclear import and drives TDP43 mislocalization in ALS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.29.635429v1?rss=1">
<title>
<![CDATA[
Phage defense and genome editing using novel retrons sourced from isolated environmental bacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.29.635429v1?rss=1</link>
<description><![CDATA[
Retrons are bacterial immune systems that protect a bacterial population against phages by killing infected hosts. Retrons typically comprise a reverse transcriptase, a template noncoding RNA that is partially reverse transcribed into RT-DNA, and a toxic effector. The reverse transcriptase, noncoding RNA, and RT-DNA complex sequester the toxic effector until triggered by phage infection, at which point the toxin is released to induce cell death. Due to their ability to produce single-stranded DNA in vivo, retrons have also been engineered to produce donor templates for genome editing in both prokaryotes and eukaryotes. However, the current repertoire of experimentally characterized retrons is limited, with most retrons sourced from clinical and laboratory strains of bacteria. To better understand retron biology and natural diversity, and to expand the current toolbox of retron-based genome editors, we developed a pipeline to isolate retrons and their bacterial hosts from a variety of environmental samples. Here, we present six of these novel retrons, each isolated from a different host bacterium. We characterize the full operon of these retrons and test their ability to defend against a panel of E. coli phages. For two of these retrons, we further unravel their mechanism of defense by identifying the phage genes responsible for triggering abortive infection. Finally, we engineer these retrons for genome editing in E. coli, demonstrating their potential use in a biotechnological application.
]]></description>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Mestre, M. R.</dc:creator>
<dc:creator>Rojas-Montero, M.</dc:creator>
<dc:creator>Shipman, S.</dc:creator>
<dc:date>2025-01-29</dc:date>
<dc:identifier>doi:10.1101/2025.01.29.635429</dc:identifier>
<dc:title><![CDATA[Phage defense and genome editing using novel retrons sourced from isolated environmental bacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.30.635804v1?rss=1">
<title>
<![CDATA[
Emergence of binding and catalysis from a designed generalist binding protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.30.635804v1?rss=1</link>
<description><![CDATA[
The evolution of binding and catalysis played a central role in the emergence of life. While natural proteins have finely tuned affinities for their primary ligands, they also bind weakly and promiscuously to other molecules, which serve as starting points for stepwise, incremental evolution of entirely new specificities. Thus, modern proteins emerged from the joint exploration of sequence and structural space. The ability of natural proteins to bind small molecule fragments in well-defined geometries has been widely evaluated using methods including crystallographic fragment screening. However, this approach had not been applied to de novo proteins. Here, we apply this method to explore the binding specificity of a de novo small molecule-binding protein ABLE. As in Nature, we found ABLE was capable of forming weak complexes, which were excellent starting points for designing entirely new functions, including a binder of a turn-on fluorophore and a highly efficient Kemp eliminase enzyme (kcat/KM = 2,200,000 M-1s-1) approaching the diffusion limit. This work illustrates how simultaneous consideration of both sequence and chemical structure diversity can guide the emergence of new function in designed proteins.
]]></description>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Bergmann, L.</dc:creator>
<dc:creator>Correy, G. J.</dc:creator>
<dc:creator>Tan, S.</dc:creator>
<dc:creator>Hou, K.</dc:creator>
<dc:creator>Biel, J.</dc:creator>
<dc:creator>Lu, L.</dc:creator>
<dc:creator>Bakanas, I.</dc:creator>
<dc:creator>Polizzi, N. F.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:date>2025-01-31</dc:date>
<dc:identifier>doi:10.1101/2025.01.30.635804</dc:identifier>
<dc:title><![CDATA[Emergence of binding and catalysis from a designed generalist binding protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.31.635826v1?rss=1">
<title>
<![CDATA[
Initial leads to combat streptogramin resistance generated from X-ray fragment screening against VatD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.31.635826v1?rss=1</link>
<description><![CDATA[
Streptogramins, a potent antibiotic class targeting bacterial ribosomes, are effective against severe infections such as those caused by vancomycin-resistant Staphylococcus aureus (VRSA) and Enterococcus faecium (VRE). The synergistic binding of group A and B streptogramins to the peptidyl transferase center (PTC) and nascent peptide exit tunnel (NPET), respectively, yields a bactericidal effect. However, the efficacy of streptogramins is compromised by resistance mechanisms, including the enzymatic acetylation of group A streptogramins by Virginiamycin Acetyl Transferase (Vat) enzymes, which reduce antibiotic affinity for the bacterial ribosome. Here, we applied fragment-based drug discovery (FBDD) to identify starting points for the development of inhibitors of VatD. X-ray crystallography screening revealed three primary fragment-binding sites on VatD. In the acetyl-CoA binding site, fragments bound to stabilize distinct conformational states in critical residues, His82 and Trp121, within the acetyl subsite. In the antibiotic binding site, two fragments were identified that form unique hydrogen/hydrophobic bonds, suggesting specific interactions that could be leveraged for competitive inhibition by disrupting streptogramin modification. Elaborations of these fragments showed weak inhibition of VatDs acetyltransferase activity, indicating the potential for further optimization. These findings establish initial hits that could restore streptogramin efficacy by targeting VatD directly, providing a structural foundation for inhibitor development against resistant bacterial strains.
]]></description>
<dc:creator>Asthana, P. -</dc:creator>
<dc:creator>Lee, S. -</dc:creator>
<dc:creator>MacDonald, C. M.</dc:creator>
<dc:creator>Seiple, I. B.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2025-02-02</dc:date>
<dc:identifier>doi:10.1101/2025.01.31.635826</dc:identifier>
<dc:title><![CDATA[Initial leads to combat streptogramin resistance generated from X-ray fragment screening against VatD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.03.636363v1?rss=1">
<title>
<![CDATA[
A scoping study of the whole-cell imaging literature: a foundational corpus, potential for data-mining and research synthesis, and a call for standardization of an emerging field 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.03.636363v1?rss=1</link>
<description><![CDATA[
The level of cellular organization bridging the mesoscale and whole-cell scale is coming into focus as a new frontier in cell biology. Great progress has been made in unraveling the complex physical and functional interconnectivity of organelles, but how the entire organelle network is spatially arranged within the cytoplasm is only beginning to be explored. Drawing on cross-disciplinary research synthesis methods, we systematically curated the whole-cell volumetric imaging literature, resulting in a corpus consisting of 89 studies and 118 image datasets. We describe the trajectory and current state of the field between 2004 and 2024. A broad characterization, or "scoping review", of bibliometrics, study design, and reporting practices shows accelerating technological development and research output. We find high variability in study design and reporting practices, including imaging modality, model organism, cellular contexts, organelles imaged, and analyses. Due to the laborious, low-throughput nature of most volumetric imaging methods, we find trends toward small sample sizes (<10 cells) and small cell types. We describe common quantitative analyses across studies, including volumetric ratios of organelles and inter-organelle contact analyses. This work establishes the initial iteration of a growing dataset of whole-cell imaging literature and data, and motivates a call for standardized whole-cell imaging study design, reporting, and data sharing practices in the context of an emerging sub-field of cell biology. Our curated dataset now provides the basis for a plethora of future aggregate and comparative analyses to reveal larger patterns and generalized hypotheses about the systems behavior and regulation of whole-cell organelle networks. More broadly, we showcase the potential of new rigorous secondary research methods to strengthen cell biologys literature review and reproducibility toolkit, create new avenues for discovery, and promote open research practices that support secondary data-reuse and integration.
]]></description>
<dc:creator>Mirvis, M.</dc:creator>
<dc:creator>Weingard, B.</dc:creator>
<dc:creator>Goodman, S.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2025-02-04</dc:date>
<dc:identifier>doi:10.1101/2025.02.03.636363</dc:identifier>
<dc:title><![CDATA[A scoping study of the whole-cell imaging literature: a foundational corpus, potential for data-mining and research synthesis, and a call for standardization of an emerging field]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.31.635980v1?rss=1">
<title>
<![CDATA[
Swirling motion of breast cancer cells radially aligns collagen fibers to enable collective invasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.31.635980v1?rss=1</link>
<description><![CDATA[
In breast cancer (BC), radial alignment of collagen fibers at the tumor-matrix interface facilitates collective invasion of cancer cells into the surrounding stromal matrix, a critical step toward metastasis. Collagen remodeling is driven by proteases and cellular forces, mediated by matrix mechanical plasticity, or irreversible matrix deformation in response to force. However, the specific mechanisms causing collagen radial alignment remain unclear. Here, we study collective invasion of BC tumor spheroids in collagen-rich matrices. Increasing plasticity to BC-relevant ranges facilitates invasion, with increasing stiffness potentiating a transition from single cell to collective invasion. At enhanced plasticity, cells radially align collagen at the tumor-matrix interface prior to invasion. Surprisingly, cells migrate tangentially to the tumor-matrix interface in a swirling-like motion, perpendicular to the direction of alignment. Mechanistically, swirling generates local shear stresses, leading to distally propagating contractile radial stresses due to negative normal stress, an underappreciated property of collagen-rich matrices. These contractile stresses align collagen fibers radially, facilitating collective invasion. The basement membrane (BM), which separates epithelia from stroma in healthy tissues, acts as a mechanical insulator by preventing swirling cells from aligning collagen. Thus, after breaching the BM, swirling of BC cells at the tumor-stroma interface radially aligns collagen to facilitate invasion.
]]></description>
<dc:creator>Saraswathibhatla, A.</dc:creator>
<dc:creator>Rabbi, M. F.</dc:creator>
<dc:creator>Varma, S.</dc:creator>
<dc:creator>Srivastava, V.</dc:creator>
<dc:creator>Ilina, O.</dc:creator>
<dc:creator>Alyafei, N. H. K.</dc:creator>
<dc:creator>Hodgson, L.</dc:creator>
<dc:creator>Gartner, Z.</dc:creator>
<dc:creator>Friedl, P.</dc:creator>
<dc:creator>West, R.</dc:creator>
<dc:creator>Kim, T. B.</dc:creator>
<dc:creator>Chaudhuri, O.</dc:creator>
<dc:date>2025-02-06</dc:date>
<dc:identifier>doi:10.1101/2025.01.31.635980</dc:identifier>
<dc:title><![CDATA[Swirling motion of breast cancer cells radially aligns collagen fibers to enable collective invasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.09.637334v1?rss=1">
<title>
<![CDATA[
A lateralized pathway for associating nutrients with flavors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.09.637334v1?rss=1</link>
<description><![CDATA[
Animals learn about the external world, in part, via interoceptive signals1,2. For example, the nutrient content of food is first estimated in the mouth, in the form of flavor, and then measured again via slower signals from the gut. How these signals from the mouth and gut are integrated to drive learning is unknown. Here we identify a lateralized dopamine pathway that is specialized for learning about the nutrient content of food. We show that dopamine neurons in the ventral tegmental area (VTADA) are necessary for associating nutrients with flavors, and that post-ingestive nutrients trigger DA release selectively in a small region of the anterior basolateral amygdala (BLA) but not canonical DA targets in striatum. Remarkably, this nutrient-triggered DA release occurs preferentially on the left side of the brain in both mice and humans, revealing that the DA system is functionally lateralized. We identify the gut sensors that are responsible for nutrient-triggered DA release; show that they activate BLA-projecting DA neurons defined by expression of cholecystokinin (CCK); and demonstrate that stimulation of DA axon terminals in the anterior BLA drives flavor-nutrient learning but not other aspects of feeding behavior. Two-photon imaging of neurons in the left anterior BLA reveals that they integrate gustatory and post-ingestive cues, and silencing these neurons prevents flavor-nutrient learning. These findings establish a neural basis for how animals learn about the nutrient content of their food. They also reveal unexpectedly that post-ingestive nutrients are differentially represented on the right and left sides of the brain.
]]></description>
<dc:creator>Grove, J. C. R.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Backes, H.</dc:creator>
<dc:creator>Kuzmanovic, B.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Ubadiah, V.</dc:creator>
<dc:creator>Qiu, L.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Small, D. M.</dc:creator>
<dc:creator>Tittgemeyer, M.</dc:creator>
<dc:creator>Knight, Z. A.</dc:creator>
<dc:date>2025-02-10</dc:date>
<dc:identifier>doi:10.1101/2025.02.09.637334</dc:identifier>
<dc:title><![CDATA[A lateralized pathway for associating nutrients with flavors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.10.637575v1?rss=1">
<title>
<![CDATA[
Mammalian oocytes receive maternal-effect RNAs from granulosa cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.10.637575v1?rss=1</link>
<description><![CDATA[
It is currently thought that growing mammalian oocytes receive only small molecules via gap junctions from surrounding support cells, the granulosa cells. From the study of chimeric preantral oocyte and granulosa cell reaggregations, we provide evidence that growing mouse oocytes receive mRNAs from granulosa cells. Among the >1,000 granulosa-transcribed RNAs we identified in the oocyte, those that contribute to proper oocyte maturation and early embryo development were highly enriched. Predicted motifs for two RNA-binding proteins that function in RNA trafficking, FMRP and TDP43, were abundant in the UTRs of the granulosa-derived transcripts. Immunostaining demonstrated that both FMRP and TDP43 co-localize with the actin-rich granulosa cell protrusions that span the zone pellucida and connect to the oocyte, suggesting their role in importing mRNAs. Our results offer the possibility that oocyte failure may not always reflect an intrinsic oocyte deficiency but could arise from insufficient supply of maternal transcripts by granulosa cells during oocyte growth.
]]></description>
<dc:creator>Munger, S. C.</dc:creator>
<dc:creator>Tijani, A.</dc:creator>
<dc:creator>Doherty, C. A.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:date>2025-02-11</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.637575</dc:identifier>
<dc:title><![CDATA[Mammalian oocytes receive maternal-effect RNAs from granulosa cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.11.637252v1?rss=1">
<title>
<![CDATA[
Human Papilloma Virus does not fully inactivate p53 in HNSCC 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.11.637252v1?rss=1</link>
<description><![CDATA[
Head and neck squamous cell carcinoma (HNSCC) is a major global health challenge. Inactivation of the tumor suppressor p53 is the most frequent molecular event in this malignancy. p53 inactivation occurs either through TP53 mutations in human papilloma virus (HPV)-negative cases or via HPV-mediated p53 degradation in HPV-positive (HPV+) cases, where most tumors retain a wild-type (WT) TP53 allele. This underscores the critical role of p53-regulated processes in HNSCC pathogenesis. Clinically, HPV+ HNSCC is associated with significantly better outcomes than HPV-negative cases. However, despite HPV E6-mediated degradation of p53, approximately 10% of HPV+ HNSCC tumors harbor TP53 mutations, suggesting an additional selective pressure to suppress p53 signaling.

In this study, we demonstrate that HPV+ TP53-WT HNSCC cells have residual, tumor suppressive p53 activity. Specifically, analysis of human tumor data reveals that among HPV+ HNSCC cases, those with WT TP53 have significantly better survival outcomes than both HPV+ cases with TP53 mutations and HPV-negative cases. Experimentally, genetic ablation of WT p53 in HPV+ HNSCC cells increased proliferation, migration, and invasion. Transcriptomic analysis revealed that p53 continues to regulate gene expression despite the presence of HPV.

Further, human tumors with HPV+ TP53-WT status exhibit tumor-suppressive methylation patterns, fewer chromosomal alterations, and suppression of PI3K-AKT signaling compared to HPV+ TP53-mutant tumors. Importantly, loss of WT p53 in HPV+ HNSCC cells increased the levels of PI3K catalytic subunit p110, reduced expression of the molecular PI3K-AKT inhibitor INPP5D and enhanced sensitivity to pharmacologic PI3K inhibition.

Together, our findings challenge the prevailing view that p53 is completely inactivated in HPV+ HNSCC and reveal tumor suppressive, p53-driven mechanisms that persist in these tumors. These insights highlight a potential role for TP53-based stratification in guiding treatment decisions and suggest new therapeutic vulnerabilities in HPV+ HNSCC.
]]></description>
<dc:creator>Gencel-Augusto, J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Woerner, L. C.</dc:creator>
<dc:creator>Borah, A. A.</dc:creator>
<dc:creator>Myers, J. N.</dc:creator>
<dc:creator>Johnson, D. E.</dc:creator>
<dc:creator>Grandis, J. R.</dc:creator>
<dc:date>2025-02-14</dc:date>
<dc:identifier>doi:10.1101/2025.02.11.637252</dc:identifier>
<dc:title><![CDATA[Human Papilloma Virus does not fully inactivate p53 in HNSCC]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.14.638232v1?rss=1">
<title>
<![CDATA[
NF-κB dependent gene expression and plasma IL-1β, TNFα and GCSF drive transcriptomic diversity and CD4:CD8 ratio in people with HIV on ART. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.14.638232v1?rss=1</link>
<description><![CDATA[
Despite antiretroviral therapy (ART), people with HIV (PWH) on ART experience higher rates of morbidity and mortality vs. age-matched HIV negative controls, which may be driven by chronic inflammation due to persistent virus. We performed bulk RNA sequencing (RNA-seq) on peripheral CD4+ T cells, as well as quantified plasma immune marker levels from 154 PWH on ART to identify host immune signatures associated with immune recovery (CD4:CD8) and HIV persistence (cell-associated HIV DNA and RNA). Using a novel dimension reduction tool - Pairwise Controlled Manifold Approximation (PaCMAP), we defined three distinct participant transcriptomic clusters. We found that these three clusters were largely defined by differential expression of genes regulated by the transcription factor NF-{kappa}B. While clustering was not associated with HIV reservoir size, we observed an association with CD4:CD8 ratio, a marker of immune recovery and prognostic factor for mortality in PWH on ART. Furthermore, distinct patterns of plasma IL-1{beta}, TNF- and GCSF were also strongly associated with the clusters, suggesting that these immune markers play a key role in CD4+ T cell transcriptomic diversity and immune recovery in PWH on ART. These findings reveal novel subgroups of PWH on ART with distinct immunological characteristics, and define a transcriptional signature associated with clinically significant immune parameters for PWH. A deeper understanding of these subgroups could advance clinical strategies to treat HIV-associated immune dysfunction.
]]></description>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Gornalusse, G.</dc:creator>
<dc:creator>Siegel, D.</dc:creator>
<dc:creator>Barbehenn, A.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Hecht, F.</dc:creator>
<dc:creator>Pilcher, C.</dc:creator>
<dc:creator>Semenova, L.</dc:creator>
<dc:creator>Murdoch, D. M.</dc:creator>
<dc:creator>Margolis, D. M.</dc:creator>
<dc:creator>Levy, C. N.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Rudin, C. D.</dc:creator>
<dc:creator>Hladik, F.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Lee, S. A.</dc:creator>
<dc:creator>Browne, E. P.</dc:creator>
<dc:date>2025-02-14</dc:date>
<dc:identifier>doi:10.1101/2025.02.14.638232</dc:identifier>
<dc:title><![CDATA[NF-κB dependent gene expression and plasma IL-1β, TNFα and GCSF drive transcriptomic diversity and CD4:CD8 ratio in people with HIV on ART.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.14.638366v1?rss=1">
<title>
<![CDATA[
Partial Input Loss Differentially Modifies Neural Pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.14.638366v1?rss=1</link>
<description><![CDATA[
Following input loss from degeneration, injury, and/or aging, downstream circuits undergo modifications that can impact neural computations. How neural computations across different pathways are affected by common input loss remain understudied. Using the retina to leverage known cell types, well-defined circuitry, and molecular tools, we show how multiple pathways adjust their functional properties differently to common input loss and further locate these changes within each pathway. Specifically, we asked if two OFF ganglion cell types, alpha OFF-sustained (AOFF-S) and OFF-transient (AOFF-T) cells, and their respective dominant presynaptic partners, type 2 and type 3a cone bipolar cells, respond differentially to partial cone loss. We find that AOFF-T ganglion cells exhibit more circuit changes than AOFF-S ganglion cells, resulting in altered spatiotemporal tuning following partial cone loss. We show that the underlying mechanisms include changes in glutamatergic, GABAergic, and glycinergic circuits in the pathway of AOFF-T ganglion cells. In response to common input loss, our study finds different locations of circuit modifications across OFF pathways. In two OFF pathways, these distinct functional changes contribute to maintaining perceptually relevant information, preserving key visual features despite input loss. These findings provide insight into how sensory systems can compensate to ultimately serve vision.
]]></description>
<dc:creator>Lee, J. Y.</dc:creator>
<dc:creator>Kastner, D. B.</dc:creator>
<dc:creator>Harris, S. C.</dc:creator>
<dc:creator>Santina, L. D.</dc:creator>
<dc:creator>John, J. V.</dc:creator>
<dc:creator>Stone, N. S.</dc:creator>
<dc:creator>Dunn, F. A.</dc:creator>
<dc:date>2025-02-16</dc:date>
<dc:identifier>doi:10.1101/2025.02.14.638366</dc:identifier>
<dc:title><![CDATA[Partial Input Loss Differentially Modifies Neural Pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.12.637954v1?rss=1">
<title>
<![CDATA[
Coordinated macrophage and T cell interactions mediate response to checkpoint blockade in colorectal cancer. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.12.637954v1?rss=1</link>
<description><![CDATA[
Mismatch repair deficiency (MMRd), either due to inherited or somatic mutation, is prevalent in colorectal cancer (CRC) and other cancers. While anti-PD-1 therapy is utilized in both local and advanced disease, up to 50% of MMRd CRC fail to respond. Using animal and human models of MMRd, we determined that interactions between MHC+ C1Q+ CXCL9+ macrophages and TCF+ BHLHE40+ PRF1+ T cell subsets are associated with control of MMRd tumor growth, during anti-PD-1 treatment. In contrast, resistance is associated with upregulation of TIM3, LAG3, TIGIT, and PD-1 expression on T cells, and infiltration of the tumor with immunosuppressive TREM2+ macrophages and monocytes. By combining anti-PD-1 with anti-LAG3/CTLA4/TREM2, up to 100% tumor eradication was achieved in MMRd CRC and remarkably, in >70% in MMRp CRC. This study identifies key T cell and macrophage subsets mediating the efficacy of immunotherapy in overcoming immune escape in both MMRd and MMRp CRC settings.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=150 SRC="FIGDIR/small/637954v1_ufig1.gif" ALT="Figure 1">
View larger version (47K):
org.highwire.dtl.DTLVardef@1778bd6org.highwire.dtl.DTLVardef@17da4e5org.highwire.dtl.DTLVardef@1d4d1d2org.highwire.dtl.DTLVardef@11b58e3_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LIAnti-PD-1 therapy leads to the accumulation and colocalization of MHCI/II+ C1Q+ CXCL9+ macrophages and DCs with TCF+ CCL5+ T cells that have high TCR diversity.
C_LIO_LIResistance to anti-PD-1 therapy involves multiple T cell checkpoints, TREM2+ macrophages, IL1B+ TREM1+ monocytes and neutrophils, and IFITM+ tumor cells.
C_LIO_LISimultaneous blockade of PD-1, LAG3, CTLA-4 and TREM2 dramatically prevents progression of both MMRd and MMRp tumors.
C_LIO_LICombination therapy completely eliminates tumors by leveraging MHC+ macrophage, CD4+ and CD8+ T cell interactions, facilitating durable anti-tumor effects.
C_LI
]]></description>
<dc:creator>Mestrallet, G.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:creator>Vaninov, N.</dc:creator>
<dc:creator>Cho, N. W.</dc:creator>
<dc:creator>Velazquez, L.</dc:creator>
<dc:creator>Ananthanarayanan, A.</dc:creator>
<dc:creator>Spitzer, M.</dc:creator>
<dc:creator>Vabret, N.</dc:creator>
<dc:creator>Cimen Bozkus, C.</dc:creator>
<dc:creator>Samstein, R. M.</dc:creator>
<dc:creator>Bhardwaj, N.</dc:creator>
<dc:date>2025-02-17</dc:date>
<dc:identifier>doi:10.1101/2025.02.12.637954</dc:identifier>
<dc:title><![CDATA[Coordinated macrophage and T cell interactions mediate response to checkpoint blockade in colorectal cancer.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.14.638277v1?rss=1">
<title>
<![CDATA[
Near 100% efficient homology-dependent genome engineering in the human fungal pathogen Cryptococcus neoformans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.14.638277v1?rss=1</link>
<description><![CDATA[
We recently described CRISPR/Cas9-based short homology-dependent genome engineering in the human fungal pathogen Cryptococcus neoformans, a haploid budding yeast that is the most common cause of fungal meningitis and an emerging model organism. This was achieved by electroporation of strains stably expressing a codon-optimized Cas9 with two separate DNA molecules, one encoding a selectable marker flanked by short homology arms and a second encoding a sgRNA under the control of the U6 snRNA promoter. However, the efficiency of desired homology-dependent repair relative to undesired non-homologous end-joining (NHEJ) events can be low and variable. Here, we describe methods and strains enabling extremely efficient ([~]99%) homology-dependent genome editing in C. neoformans. This high-efficiency method requires two manipulations. First, we placed the sgRNA-expressing segment into the marker-containing DNA flanked by targeting homology; thus, only a single DNA molecule is introduced into cells. Second, we used a strain mutant for the non-homologous end-joining factor Ku80 (encoded by YKU80). We also report the engineering of a yku80::amdS mutant strain harboring an insertion mutation that can be removed scarlessly via recombination between direct repeats. This enables the functional restoration of YKU80 after homology- dependent genome editing via selection against the amdS marker using fluoroacetamide. This approach minimizes documented drawbacks of using Ku-defective strains in downstream experiments. Finally, we describe a plasmid series that enables rapid cloning of sgRNA-marker constructs for genomic manipulation of C. neoformans, including gene deletion and C-terminal tagging. These methods, strains, and plasmids accelerate the genomic manipulation of C. neoformans.
]]></description>
<dc:creator>Nalley, M.</dc:creator>
<dc:creator>Banerjee, S.</dc:creator>
<dc:creator>Huang, M. Y.</dc:creator>
<dc:creator>Madhani, H. D.</dc:creator>
<dc:date>2025-02-18</dc:date>
<dc:identifier>doi:10.1101/2025.02.14.638277</dc:identifier>
<dc:title><![CDATA[Near 100% efficient homology-dependent genome engineering in the human fungal pathogen Cryptococcus neoformans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.18.638918v1?rss=1">
<title>
<![CDATA[
Genome modeling and design across all domains of life with Evo 2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.18.638918v1?rss=1</link>
<description><![CDATA[
All of life encodes information with DNA. While tools for sequencing, synthesis, and editing of genomic code have transformed biological research, intelligently composing new biological systems would also require a deep understanding of the immense complexity encoded by genomes. We introduce Evo 2, a biological foundation model trained on 9.3 trillion DNA base pairs from a highly curated genomic atlas spanning all domains of life. We train Evo 2 with 7B and 40B parameters to have an unprecedented 1 million token context window with single-nucleotide resolution. Evo 2 learns from DNA sequence alone to accurately predict the functional impacts of genetic variation--from noncoding pathogenic mutations to clinically significant BRCA1 variants--without task-specific finetuning. Applying mechanistic interpretability analyses, we reveal that Evo 2 autonomously learns a breadth of biological features, including exon-intron boundaries, transcription factor binding sites, protein structural elements, and prophage genomic regions. Beyond its predictive capabilities, Evo 2 generates mitochondrial, prokaryotic, and eukaryotic sequences at genome scale with greater naturalness and coherence than previous methods. Guiding Evo 2 via inference-time search enables controllable generation of epigenomic structure, for which we demonstrate the first inference-time scaling results in biology. We make Evo 2 fully open, including model parameters, training code, inference code, and the OpenGenome2 dataset, to accelerate the exploration and design of biological complexity.
]]></description>
<dc:creator>Brixi, G.</dc:creator>
<dc:creator>Durrant, M. G.</dc:creator>
<dc:creator>Ku, J.</dc:creator>
<dc:creator>Poli, M.</dc:creator>
<dc:creator>Brockman, G.</dc:creator>
<dc:creator>Chang, D.</dc:creator>
<dc:creator>Gonzalez, G. A.</dc:creator>
<dc:creator>King, S. H.</dc:creator>
<dc:creator>Li, D. B.</dc:creator>
<dc:creator>Merchant, A. T.</dc:creator>
<dc:creator>Naghipourfar, M.</dc:creator>
<dc:creator>Nguyen, E.</dc:creator>
<dc:creator>Ricci-Tam, C.</dc:creator>
<dc:creator>Romero, D. W.</dc:creator>
<dc:creator>Sun, G.</dc:creator>
<dc:creator>Taghibakshi, A.</dc:creator>
<dc:creator>Vorontsov, A.</dc:creator>
<dc:creator>Yang, B.</dc:creator>
<dc:creator>Deng, M.</dc:creator>
<dc:creator>Gorton, L.</dc:creator>
<dc:creator>Nguyen, N.</dc:creator>
<dc:creator>Wang, N. K.</dc:creator>
<dc:creator>Adams, E.</dc:creator>
<dc:creator>Baccus, S. A.</dc:creator>
<dc:creator>Dillmann, S.</dc:creator>
<dc:creator>Ermon, S.</dc:creator>
<dc:creator>Guo, D.</dc:creator>
<dc:creator>Ilango, R.</dc:creator>
<dc:creator>Janik, K.</dc:creator>
<dc:creator>Lu, A. X.</dc:creator>
<dc:creator>Mehta, R.</dc:creator>
<dc:creator>Mofrad, M. R. K.</dc:creator>
<dc:creator>Ng, M. Y.</dc:creator>
<dc:creator>Pannu, J.</dc:creator>
<dc:creator>Re, C.</dc:creator>
<dc:creator>Schmok, J. C.</dc:creator>
<dc:creator>St. John, J.</dc:creator>
<dc:creator>Sullivan, J.</dc:creator>
<dc:creator>Zhu, K.</dc:creator>
<dc:creator>Zynda, G.</dc:creator>
<dc:creator>Balsam, D.</dc:creator>
<dc:creator>Collison, P.</dc:creator>
<dc:creator>Costa, A. B.</dc:creator>
<dc:creator>Hernandez-Boussard, T.</dc:creator>
<dc:creator>Ho, E.</dc:creator>
<dc:creator>Liu, M.-Y.</dc:creator>
<dc:creator>McGrath, T.</dc:creator>
<dc:creator>P</dc:creator>
<dc:date>2025-02-21</dc:date>
<dc:identifier>doi:10.1101/2025.02.18.638918</dc:identifier>
<dc:title><![CDATA[Genome modeling and design across all domains of life with Evo 2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.24.639979v1?rss=1">
<title>
<![CDATA[
A network of coiled-coil and actin-like proteins controls the cellular organization of magnetosome organelles in deep-branching magnetotactic bacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.24.639979v1?rss=1</link>
<description><![CDATA[
Magnetotactic Bacteria (MTB) are a diverse group of microorganisms that use magnetosomes, organelles composed of magnetite or greigite, to navigate along geomagnetic fields. While MTB span several phyla and exhibit diverse phenotypes, magnetosome formation has been mechanistically studied in only two species of Alphaproteobacteria. Here, we use Desulfovibrio magneticus RS-1 to uncover the mechanisms behind tooth-shaped magnetosome assembly in deep-branching MTB. Our findings show that magnetic particles in RS-1 initially form randomly within the cell before localizing to the positive cell curvature. Genetic and proteomic analyses indicate that early biomineralization involves membrane-associated proteins found in all MTB, while later stages depend on coiled-coil (Mad20, 23, 25, and 26) and actin-like (MamK and Mad28) proteins, most of which are unique to deep-branching MTB. These findings suggest that while biomineralization originates from a common ancestor, magnetosome chain organization has distinct evolutionarily origins among different MTB lineages.
]]></description>
<dc:creator>Russell, V. V.</dc:creator>
<dc:creator>Iavarone, A. T.</dc:creator>
<dc:creator>Ozyamak, E.</dc:creator>
<dc:creator>Grant, C.</dc:creator>
<dc:creator>Komeili, A.</dc:creator>
<dc:date>2025-02-24</dc:date>
<dc:identifier>doi:10.1101/2025.02.24.639979</dc:identifier>
<dc:title><![CDATA[A network of coiled-coil and actin-like proteins controls the cellular organization of magnetosome organelles in deep-branching magnetotactic bacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.24.639882v1?rss=1">
<title>
<![CDATA[
Natural Killer Cell Receptor Signaling and Activation Depend on Cell Cycle Stages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.24.639882v1?rss=1</link>
<description><![CDATA[
Receptor signaling in Natural Killer (NK) cells leads to post-translational modification (e.g., phosphorylation) of sub-cellular signaling proteins within minutes of receptor stimulation that eventually give rise to diverse effector functions including cell proliferation. Recent single-cell mass cytometry (i.e., CyTOF) experiments in macrophages showed variations of abundances of phosphorylated signaling proteins across cell cycle states indicating a dependence of cell signaling kinetics on an order of magnitude slower kinetics (~ several hours) of cell cycle transitions. We investigated cell cycle dependence of NKG2D signaling kinetics in NK cells by CyTOF measurements performed on IL-2-treated NKG2D-stimulated primary human CD56dim NK cells. The CyTOF experiments revealed monotonic or semi-monotonic increases of the average protein abundances of the majority of signaling proteins such as pCrkL, pPLC{gamma}2, and pErk, and the degranulation marker protein CD107a with progressing cell cycle states at specific time points post-NKG2D stimulation; however, several proteins such as pVav1, pS6, and pAkt, and early activation marker protein CD69 also showed non-monotonic variations in the average abundances with progressing cell cycle states. We used minimal mathematical and computational models coupling signaling and cell cycle processes to show that non-monotonic variations in the signaling protein abundances with progressing cell cycle stages are likely to arise in situations where protein synthesis and degradation and signaling kinetics are actively regulated by cell cycle processes.
]]></description>
<dc:creator>Wethington, D.</dc:creator>
<dc:creator>Nayak, I.</dc:creator>
<dc:creator>Jensen, H.</dc:creator>
<dc:creator>Stewart, W. C.</dc:creator>
<dc:creator>Aguilar, O. A.</dc:creator>
<dc:creator>Chen, S.-Y.</dc:creator>
<dc:creator>Nolan, G. P.</dc:creator>
<dc:creator>Behbehani, G. K.</dc:creator>
<dc:creator>Lanier, L. L.</dc:creator>
<dc:creator>Das, J.</dc:creator>
<dc:date>2025-02-28</dc:date>
<dc:identifier>doi:10.1101/2025.02.24.639882</dc:identifier>
<dc:title><![CDATA[Natural Killer Cell Receptor Signaling and Activation Depend on Cell Cycle Stages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.27.640481v1?rss=1">
<title>
<![CDATA[
Comparative analysis of human and mouse ovaries across age 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.27.640481v1?rss=1</link>
<description><![CDATA[
Mouse is a tractable model for human ovarian biology, however its utility is limited by incomplete understanding of how transcription and signaling differ interspecifically and with age. We compared ovaries between species using 3D-imaging, single-cell transcriptomics, and functional studies. In mice, we mapped declining follicle numbers and oocyte competence during aging; in human ovaries, we identified cortical follicle pockets and density changes. Oocytes had species-specific gene expression patterns during growth that converged toward maturity. Age-related transcriptional changes were greater in oocytes than granulosa cells across species, although mature oocytes change more in humans. We identified ovarian sympathetic nerves and glia; nerve density increased in aged human ovaries and, when ablated in mice, perturbed folliculogenesis. This comparative atlas defines shared and species-specific hallmarks of ovarian biology.
]]></description>
<dc:creator>Gaylord, E. A.</dc:creator>
<dc:creator>Foecke, M. F.</dc:creator>
<dc:creator>Samuel, R. M.</dc:creator>
<dc:creator>Soygur, B.</dc:creator>
<dc:creator>Detweiler, A. M.</dc:creator>
<dc:creator>Dorman, L.</dc:creator>
<dc:creator>Borja, M.</dc:creator>
<dc:creator>Laird, A. E.</dc:creator>
<dc:creator>Arjyal, R.</dc:creator>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Neff, N.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:date>2025-03-03</dc:date>
<dc:identifier>doi:10.1101/2025.02.27.640481</dc:identifier>
<dc:title><![CDATA[Comparative analysis of human and mouse ovaries across age]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.27.640483v1?rss=1">
<title>
<![CDATA[
Single-cell RNA sequencing of peripheral blood defines two immunological subtypes of Sjögren's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.27.640483v1?rss=1</link>
<description><![CDATA[
Sjogrens disease (SjD) is a heterogeneous autoimmune disorder with significant clinical and molecular diversity. While anti-SSA antibodies serve as a hallmark serological biomarker, nearly half of patients lack them, raising fundamental questions about distinct pathogenic mechanisms. To address this, we performed single-cell RNA sequencing with surface protein profiling on 1.5 million PBMCs from 333 participants in the Sjogrens International Collaborative Clinical Alliance cohort, comparing those with and without SjD and stratifying them by anti-SSA status. Our analysis identified two immunologically distinct subtypes of SjD, with SSA-positive patients exhibiting a dominant and persistent IFN-I signature, profoundly impacting immune cell maturation. Notably, transitional B cells were disproportionately affected, displaying signs of premature maturation, reduced BCR diversity, and shorter CDR3 regions, reinforced interactions with proinflammatory cells, thereby fostering an environment conducive to autoreactivity. By contrast, SSA-negative SjD participants lacked an upregulated IFN-I signature, challenging prevailing pathogenic models and suggesting alternative immune dysregulation pathways. These findings support a two-disease model of SjD and highlight transitional B cells as both a key biomarker and a potential therapeutic target. Targeting IFN-I signaling and transitional B cell maturation may represent a novel therapeutic avenue to modulate immune dysregulation and prevent autoreactivity in SjD.
]]></description>
<dc:creator>Urbanski, G.</dc:creator>
<dc:creator>Taylor, K. E.</dc:creator>
<dc:creator>Flynn, E.</dc:creator>
<dc:creator>Norouzi, A.</dc:creator>
<dc:creator>Chu, C.</dc:creator>
<dc:creator>Davidson, B.</dc:creator>
<dc:creator>Ghosh, A.</dc:creator>
<dc:creator>Nititham, J.</dc:creator>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Poon, A. W.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Combes, A. K.</dc:creator>
<dc:creator>Eckalbar, W. L.</dc:creator>
<dc:creator>Criswell, L. A.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Shiboski, C. H.</dc:creator>
<dc:date>2025-03-03</dc:date>
<dc:identifier>doi:10.1101/2025.02.27.640483</dc:identifier>
<dc:title><![CDATA[Single-cell RNA sequencing of peripheral blood defines two immunological subtypes of Sjögren's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.28.640798v1?rss=1">
<title>
<![CDATA[
Independent signaling pathways provide a fail-safe mechanism to prevent tumorigenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.28.640798v1?rss=1</link>
<description><![CDATA[
Controlled signaling activity is vital for normal tissue homeostasis and oncogenic signaling activation facilitates tumorigenesis. Here we use single-cell transcriptomics to investigate the effects of pro-proliferative signaling on epithelial homeostasis using the Drosophila follicle cell lineage. Notably, EGFR-Ras overactivation induces cell cycle defects by activating the transcription factors Pointed and E2f1 and impedes differentiation. Hh signaling simultaneously promotes an undifferentiated state and induces differentiation via activation of EMT-associated transcription factors zfh1 and Mef2. As a result, overactivation of Hh signaling generates a transcriptional hybrid state comparable to epithelial-mesenchymal-transition. Co-overactivation of Hh signaling with EGFR-Ras signaling blocks differentiation and induces key characteristics of tumor cells including a loss of tissue architecture caused by reduced expression of cell adhesion molecules, sustained proliferation and an evasion of cell cycle checkpoints. These findings provide new insight into how non-interacting signaling pathways converge at the transcriptional level to prevent malignant cell behavior.

Graphical abstract

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]]></description>
<dc:creator>Rust, K.</dc:creator>
<dc:creator>Schubert, A.</dc:creator>
<dc:creator>Peralta, J. M.</dc:creator>
<dc:creator>Nystul, T. G.</dc:creator>
<dc:date>2025-03-06</dc:date>
<dc:identifier>doi:10.1101/2025.02.28.640798</dc:identifier>
<dc:title><![CDATA[Independent signaling pathways provide a fail-safe mechanism to prevent tumorigenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.04.641363v1?rss=1">
<title>
<![CDATA[
Searching for influencers among placental immune cells in preeclampsia. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.04.641363v1?rss=1</link>
<description><![CDATA[
Cells in maternal and fetal immune systems may communicate, leading to immune tolerance during pregnancy; however, this hypothesis remains controversial. Here, we profiled single-cell transcriptional signatures in placental layers comprising the maternal-fetal interface and deep placenta, then searched for genes associated with preeclampsia. To investigate the underlying principle of the failure of immune tolerance, we started by clarifying the systemic framework, comprising models of immune interaction frequency (IIF) and specific triggers (i.e., influencers) of tolerance (IT). We generated single-cell transcriptional profiles of normal term (Norms) and preeclampsia preterm (PePT) parturitions. Fetal and maternal cells are admixed across the placenta, for both Norms and PePTs, rejecting the IIF model of immune failure during pregnancy posed by excessive interactions between fetomaternal cells. Whereas placental layers are well mixed with maternal cells, we identified a conserved gradual immune transition of fetal T-cells in both PePT and Norm, disproving the IIF model. To search for influencers of PePT in the IT model, we established and validated a classification model for PePT and Norm immune cells, including T-cells, and then prioritized major contributors to the classifier model, which are highly enriched in ligands and receptors (p = 5.98e-5). Among the prioritized ligand receptors, SPP1 and CD44 are suggested as influencers of inflammation signatures and were experimentally validated by the exclusive colocalization of SPP1- and CD44-expressing cells in the PePT placentas. Different interleukin-4 and interferon-{psi} levels in the serum and urine of PePTs further support the contribution of SPP1 to associated pathways, including allograft rejection. Our findings provide insight into the influence of specific immune interactions between cells in the human placenta and their influencer-derived impact on PePT.
]]></description>
<dc:creator>Paik, H.</dc:creator>
<dc:creator>Ko, T. L.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Park, J.-E.</dc:creator>
<dc:creator>Bunis, D.</dc:creator>
<dc:creator>Lee, S.-K.</dc:creator>
<dc:creator>Heo, H.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.04.641363</dc:identifier>
<dc:title><![CDATA[Searching for influencers among placental immune cells in preeclampsia.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.04.641443v1?rss=1">
<title>
<![CDATA[
Assigning Targetable Molecular Pathways to Transdiagnostic Subgroups Across Autism and Related Neurodevelopmental Disorders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.04.641443v1?rss=1</link>
<description><![CDATA[
Significant genetic, behavioural and neuroanatomic heterogeneity is common in autism spectrum- and related- neurodevelopmental disorders (NDDs). This heterogeneity constrains the development of effective therapies for diverse patients in precision medicine paradigms. This has led to the search for subgroups of individuals having common etiologic factors/biology (e.g., genetic pathways), thus creating potential uniformity in prognosis and/or treatment response. Despite NDDs having a strong genetic component, only ~15-20% of individuals will present with a specific rare genetic variant considered clinically pathogenic, and therefore, subtyping efforts tend to focus on using clinical, cognitive, and/or brain imaging phenotypes to group individuals. Here we delineated mechanisms via mouse to human translational neuroscience. Using MRI derived structural neuroanatomy and a spatial transcriptomic comparison, we linked subgroups of 135 NDD relevant mouse models (3,515 individual mice) separately to two human databases, with 1,234 and 1,015 human individuals with NDDs, composed of autism, attention-deficit/hyperactivity disorder (ADHD), obsessive compulsive disorder (OCD), other related NDDs, and typically developing controls. Subgroups were significantly linked by consistent neuroanatomy across all three datasets, mouse and human, indicating that direct cross-species subgrouping and translation is consistent and reproducible. Ultimately, four specific neuroanatomical clusters were found and linked to precise molecular mechanisms: two showing a chromatin/transcription motif, with one of those showing specific links to G-protein coupled receptors (GPCR) and Notch signalling, and another two being mainly synaptic in origin, with one off those showing specific connections to axon guidance and Wnt signaling. Assigning molecular pathways, and thus genetic information, from the mouse to individual participants provides an insight into undetected and/or related genetic variants that could be working in combination or interacting with an environmental influence. Moreover, the subgroups found are transdiagnostic, including participants with autism, ADHD, and OCD, which indicates that NDDs as a whole can be subdivided into consistent neuroanatomical clusters with cohesive underlying biological mechanisms. This work allows us to bridge the gap between preclinical models and human disorders, linking previously idiopathic human patients to pertinent genetics, molecular mechanisms, and pathways.
]]></description>
<dc:creator>Ellegood, J.</dc:creator>
<dc:creator>Beauchamp, A.</dc:creator>
<dc:creator>Yee, Y.</dc:creator>
<dc:creator>Devenyi, G.</dc:creator>
<dc:creator>Ziolkowski, J.</dc:creator>
<dc:creator>Qiu, L.</dc:creator>
<dc:creator>Askalan, R.</dc:creator>
<dc:creator>Ayub, M.</dc:creator>
<dc:creator>Suetterlin, P.</dc:creator>
<dc:creator>Donovan, A.</dc:creator>
<dc:creator>Basson, M. A.</dc:creator>
<dc:creator>Quesnel, K. M.</dc:creator>
<dc:creator>Berube, N. G.</dc:creator>
<dc:creator>Woo, T.</dc:creator>
<dc:creator>Beversdorf, D.</dc:creator>
<dc:creator>Bjornsson, H.</dc:creator>
<dc:creator>Blakely, R.</dc:creator>
<dc:creator>Crawley, J.</dc:creator>
<dc:creator>Crosbie, J.</dc:creator>
<dc:creator>Orr, B. O.</dc:creator>
<dc:creator>Davis, G. W.</dc:creator>
<dc:creator>Genestine, M.</dc:creator>
<dc:creator>DiCicco-Bloom, E.</dc:creator>
<dc:creator>Egan, S.</dc:creator>
<dc:creator>Fink, K. D.</dc:creator>
<dc:creator>Asbury, S.</dc:creator>
<dc:creator>Lai, J.</dc:creator>
<dc:creator>Rilett, K.</dc:creator>
<dc:creator>Foster, J. A.</dc:creator>
<dc:creator>Vincent, J. B.</dc:creator>
<dc:creator>Frankland, P.</dc:creator>
<dc:creator>Georgiades, S.</dc:creator>
<dc:creator>Penagarikano, O.</dc:creator>
<dc:creator>Geschwind, D.</dc:creator>
<dc:creator>Giger, R. J.</dc:creator>
<dc:creator>Markx, S.</dc:creator>
<dc:creator>Gogos, J.</dc:creator>
<dc:creator>Golzio, C.</dc:creator>
<dc:creator>Pagani, M.</dc:creator>
<dc:creator>Gozzi, A.</dc:creator>
<dc:creator>Pacey, L. K.</dc:creator>
<dc:creator>Hampson, D.</dc:creator>
<dc:creator>Huang, T.-N.</dc:creator>
<dc:creator>Yen, T.-L.</dc:creator>
<dc:creator>Hsueh, Y.-P.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.04.641443</dc:identifier>
<dc:title><![CDATA[Assigning Targetable Molecular Pathways to Transdiagnostic Subgroups Across Autism and Related Neurodevelopmental Disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.05.640882v1?rss=1">
<title>
<![CDATA[
In silico discovery of nanobody binders to a G-protein coupled receptor using AlphaFold-Multimer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.05.640882v1?rss=1</link>
<description><![CDATA[
Antibodies are central mediators of the adaptive immune response, and they are powerful research tools and therapeutics. Antibody discovery requires substantial experimental effort, such as immunization campaigns or in vitro library screening. Predicting antibody-antigen binding a priori remains challenging. However, recent machine learning methods raise the possibility of in silico antibody discovery, bypassing or reducing initial experimental bottlenecks. Here, we report a virtual screen using AlphaFold-Multimer (AF-M) that prospectively identified nanobody binders to MRGPRX2, a G protein-coupled receptor (GPCR) and therapeutic target for the treatment of pseudoallergic inflammation and itch. Using previously reported nanobody-GPCR structures, we identified a set of AF-M outputs that effectively discriminate between interacting and non-interacting nanobody-GPCR pairs. We used these outputs to perform a prospective in silico screen, identified nanobodies that bind MRGPRX2 with high affinity, and confirmed activity in signaling and functional cellular assays. Our results provide a proof of concept for fully computational antibody discovery pipelines that can circumvent laboratory experiments.
]]></description>
<dc:creator>Harvey, E. P.</dc:creator>
<dc:creator>Smith, J. S.</dc:creator>
<dc:creator>Hurley, J. D.</dc:creator>
<dc:creator>Granados, A.</dc:creator>
<dc:creator>Schmid, E. W.</dc:creator>
<dc:creator>Liang-Lin, J. G.</dc:creator>
<dc:creator>Paul, S.</dc:creator>
<dc:creator>Meara, E. M.</dc:creator>
<dc:creator>Ferguson, M. P.</dc:creator>
<dc:creator>Calvillo-Miranda, V. G.</dc:creator>
<dc:creator>Marks, D. S.</dc:creator>
<dc:creator>Walter, J. C.</dc:creator>
<dc:creator>Kruse, A. C.</dc:creator>
<dc:creator>Susa, K. J.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.05.640882</dc:identifier>
<dc:title><![CDATA[In silico discovery of nanobody binders to a G-protein coupled receptor using AlphaFold-Multimer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.05.641592v1?rss=1">
<title>
<![CDATA[
Revealing Changes in Linear and Nonlinear Functional Connectivity After Psilocybin and Escitalopram Treatment in Patients with Depression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.05.641592v1?rss=1</link>
<description><![CDATA[
Major Depressive Disorder (MDD) is typically characterized by altered linear functional connectivity (FC) across large-scale brain networks. Yet, it is unclear whether similar alterations are observed when nonlinear FC is examined. This study investigated how antidepressant treatment (i.e., psilocybin and escitalopram) modulates both linear FC and nonlinear FC in individuals with MDD. Here, we focused specifically on five key canonical brain networks: the Default Mode Network (DMN), Frontoparietal Network (FPN), Salience Network (SAL), Dorsal Attention Network (DAN), and Ventral Attention Network (VAN). Across both treatments, using resting-state fMRI data, we first compared changes in linear and nonlinear FC between responders and non-responders. Responders exhibited increased linear FC within the VAN and greater nonlinear FC within the DMN and VAN than non-responders. We also observed more between-network linear FC for DMN-DAN and nonlinear FC for DMN-VAN in responders than non-responders. Next, we compared treatments and observed that Psilocybin responders showed greater connectivity between FPN-VAN (linear FC), DMN-VAN (nonlinear FC), and SAL-VAN (nonlinear FC) integration than Escitalopram responders, reflecting enhanced coordination and integration between higher-order networks. Conversely, Escitalopram responders exhibited reduced within-network linear FC within the DMN and SAL and between the DMN and VAN, consistent with a dampening of self-referential and salience processing and altered attentional control. These findings highlight potentially distinct mechanisms of action for psilocybin and escitalopram. Incorporating both linear and nonlinear FC analyses provided a novel characterization of these effects, emphasizing the role of these different interactions in antidepressant response. Future studies should investigate the long-term stability of these network changes and their relationship to clinical outcomes.
]]></description>
<dc:creator>Quah, S. K. L.</dc:creator>
<dc:creator>Glick, C.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Pasquini, L.</dc:creator>
<dc:creator>Carhart-Harris, R. L.</dc:creator>
<dc:creator>Saggar, M.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.05.641592</dc:identifier>
<dc:title><![CDATA[Revealing Changes in Linear and Nonlinear Functional Connectivity After Psilocybin and Escitalopram Treatment in Patients with Depression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.06.641890v1?rss=1">
<title>
<![CDATA[
Heterocellular and homocellular electrically coupled networks of the thalamus and cortex revealed by focal photomapping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.06.641890v1?rss=1</link>
<description><![CDATA[
Electrical synapses are expressed ubiquitously across the brain and are crucial components of active neural circuitry and connectomes. Identification of coupled networks in living tissue is limited by technical demands of multiplexed recordings, and no dyes, fluorescent reporters, or genetic labels can currently fill the gap. We introduce a novel method of identifying and quantifying electrical synapses, opto-{delta}L, that combines focal photostimulation of soma-targeted opsins with a spike timing-based computation for the strength of electrical synapses to rapidly measure and map electrically coupled networks in vitro. We leverage opto-{delta}L to show that coupled networks of the mature thalamic reticular nucleus extend as far as 100 m, synapse promiscuously across genetic subtypes of neurons, and couple 1-4 neighboring neurons to each recorded hub cell. We also demonstrate application of opto-{delta}L to cortical networks. These results highlight the broad potential of opto-{delta}L to interrogate the identity and roles of electrical synapses in circuitry, behavior, and cognition.

TeaserElectrically coupled neural networks are newly unmasked by functional photomapping.
]]></description>
<dc:creator>Vaughn, M. J.</dc:creator>
<dc:creator>Bender, K.</dc:creator>
<dc:creator>Haas, J.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.06.641890</dc:identifier>
<dc:title><![CDATA[Heterocellular and homocellular electrically coupled networks of the thalamus and cortex revealed by focal photomapping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.11.642707v1?rss=1">
<title>
<![CDATA[
Tau-Associated Neuronal Loss in the Intermediate Nucleus of the Human Hypothalamus (VLPO Analog): Unveiling the Basis of NREM Sleep Dysfunction in PSP and Alzheimer's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.11.642707v1?rss=1</link>
<description><![CDATA[
Sleep disturbances are prevalent in Alzheimers disease (AD) and Progressive Supranuclear Palsy (PSP), often exacerbating disease progression. Understanding the neuropathological basis of these disturbances is essential for identifying potential therapeutic targets. This study investigates the intermediate nucleus (IntN) of the human hypothalamus--a key sleep-regulating region analogous to the rodent ventrolateral preoptic area (VLPO)--to assess neuronal loss and tau pathology in AD and PSP. Using postmortem brain tissue, we applied unbiased stereology to quantify galanin-expressing neurons and phosphorylated tau (p-tau) accumulation.

Among 26 cases analyzed, both AD and PSP exhibited significant neuronal loss in the IntN, with PSP showing the most pronounced reduction (84.9% fewer neurons than healthy controls [HC]). In AD, neuronal loss correlated with Braak staging, with late-stage AD cases (Braak 5-6) demonstrating a 76.9% reduction in galanin-expressing neurons compared to HC, while non-galanin neurons exhibited a more moderate decline (45.7%). In PSP, extensive neuronal loss precluded a clear assessment of p-tau burden. These findings suggest a differential neuronal vulnerability to tau pathology across diseases, aligning with distinct sleep disturbances observed in each condition. PSP, characterized by severe insomnia despite preserved wake-promoting neurons, may be explained by the near-total loss of NREM sleep-regulating neurons. In contrast, AD exhibits a progressive decline in both wake- and sleep-promoting neurons, contributing to excessive daytime sleepiness and sleep fragmentation.

This study provides critical insights into the selective neuronal vulnerabilities underlying sleep dysfunction in tauopathies, emphasizing the need for targeted interventions to mitigate sleep disturbances in these disorders.
]]></description>
<dc:creator>Rastegar-Pouyani, S.</dc:creator>
<dc:creator>Lew, C.</dc:creator>
<dc:creator>Pereira, F.</dc:creator>
<dc:creator>Satpati, A.</dc:creator>
<dc:creator>Paes, V.</dc:creator>
<dc:creator>Leite, R. P.</dc:creator>
<dc:creator>Suemoto, C.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Walsh, C.</dc:creator>
<dc:creator>Neylan, T.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:date>2025-03-13</dc:date>
<dc:identifier>doi:10.1101/2025.03.11.642707</dc:identifier>
<dc:title><![CDATA[Tau-Associated Neuronal Loss in the Intermediate Nucleus of the Human Hypothalamus (VLPO Analog): Unveiling the Basis of NREM Sleep Dysfunction in PSP and Alzheimer's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.12.642800v1?rss=1">
<title>
<![CDATA[
IL-6 underlies microenvironment immunosuppression and resistance to therapy in glioblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.12.642800v1?rss=1</link>
<description><![CDATA[
The glioblastoma tumor immune microenvironment (TIME) is an immunosuppressive barrier to therapy that encumbers glioblastoma responses to immune checkpoint inhibition (ICI). Immunosuppressive cytokines, pro-tumor myeloid cells, and exhausted T-cells are hallmarks of the glioblastoma TIME. Here we integrate spatial and single-cell analyses of patient-matched human glioblastoma samples before and after ICI with genetic, immunologic, single-cell, and pharmacologic studies in preclinical models to reveal that interleukin-6 (IL-6) inhibition reprograms the glioblastoma TIME to sensitize mouse glioblastoma to ICI and radiotherapy. Rare human glioblastoma patients who achieve clinical responses to ICI have lower pre-treatment IL-6 levels compared to glioblastomas who do not respond to ICI. Immune stimulatory gene therapy suppresses IL-6 tumor levels in preclinical murine models of glioblastoma. Furthermore, survival was longer in Il-6 knockout mice with orthotopic SB28 glioblastoma relative to wild-type mice. IL-6 blockade with a neutralizing antibody transiently sensitizes mouse glioblastoma to anti-PD-1 by increasing MHCII+ monocytes, CD103+ migratory dendritic cells (DCs), CD11b+ conventional DCs, and effector CD8+ T cells, and decreasing immunosuppressive Tregs. To translate these findings to a combination treatment strategy for recurrent glioblastoma patients, we show that IL-6 blockade plus ICI durably sensitizes mouse glioblastoma to high-dose radiotherapy.
]]></description>
<dc:creator>Young, J.</dc:creator>
<dc:creator>Cho, N. W.</dc:creator>
<dc:creator>Lucas, C.-H. G.</dc:creator>
<dc:creator>Najem, H.</dc:creator>
<dc:creator>Mirchia, K.</dc:creator>
<dc:creator>Chen, W. C.</dc:creator>
<dc:creator>Seo, K.</dc:creator>
<dc:creator>Zakimi, N.</dc:creator>
<dc:creator>Daggubati, V.</dc:creator>
<dc:creator>Casey-Clyde, T.</dc:creator>
<dc:creator>Nguyen, M. P.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>Phillips, J. J.</dc:creator>
<dc:creator>Ozawa, T.</dc:creator>
<dc:creator>Aghi, M.</dc:creator>
<dc:creator>Taylor, J.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Bhaduri, A.</dc:creator>
<dc:creator>Berger, M. S.</dc:creator>
<dc:creator>Heimberger, A. B.</dc:creator>
<dc:creator>Butowski, N.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:date>2025-03-14</dc:date>
<dc:identifier>doi:10.1101/2025.03.12.642800</dc:identifier>
<dc:title><![CDATA[IL-6 underlies microenvironment immunosuppression and resistance to therapy in glioblastoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.17.643721v1?rss=1">
<title>
<![CDATA[
Representing Mitochondrial Dynamics with Abstract Algebra 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.17.643721v1?rss=1</link>
<description><![CDATA[
This paper addresses the increasing need for comprehensive mathematical descriptions of cell organization by examining the algebraic structure of mitochondrial network dynamics. Mitochondria are cellular structures involved in metabolism that take the form of a network of membrane-based tubes that undergo continuous re-arrangement by a set of morphological processes, including fission and fusion, carried out by protein-based machinery. Because of their network structure, mitochondria can be represented as graphs, and the morphological operations that take place in the cell, referred to as mitochondrial dynamics, can be represented by changes to the graphs. Prior studies have classified mitochondrial graphs based on graph-theoretic features, but an alternative approach is to focus not on the graphs themselves but on the set of morphological operations inducing mitochondrial dynamics, since this may provide a simpler representation. Moreover, the operations are what determine the graphs that will be generated in a biological system. Here we show that mitochondrial dynamics constitute a groupoid that includes the automorphism group of each mitochondria graph. For multi-component mitochondria we define a graph structure that encapsulates the structure of mitochondrial dynamics. Using one of the morphological operations we can define an equivalence relation among mitochondrial graphs that allows us to replace the complex structure of the full groupoid with a vastly simpler groupoid representation based on equivalence classes. Using these formalisms we define a distance metric for similarity between mitochondrial structures based on an edit distance. In the course of defining these structures we provide a mathematical motivation for new experimental questions regarding mitochondrial fusion, the impacts of cell division on mitochondrial morphology, and the presence of a single giant component in some cell types. This work points to a general strategy for formulating a cell structure state-space, based not on the shapes of cellular structures, but on relations between the dynamic operations that produce them.
]]></description>
<dc:creator>Mostov, R.</dc:creator>
<dc:creator>Lewis, G. R.</dc:creator>
<dc:creator>Sturm, G.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2025-03-18</dc:date>
<dc:identifier>doi:10.1101/2025.03.17.643721</dc:identifier>
<dc:title><![CDATA[Representing Mitochondrial Dynamics with Abstract Algebra]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.17.643814v1?rss=1">
<title>
<![CDATA[
Stochastically Emergent Tumors offer in vivo whole genome interrogation of cancer evolution from non-malignant precursors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.17.643814v1?rss=1</link>
<description><![CDATA[
Interrogating the stochastic events underlying tumor evolution from non-malignant precursors is crucial for understanding therapy resistance. Current methods are complicated by chromosomal instability, obscuring driver identification and yielding non-representative genetics. Inspired by patient tumors that evolve without chromosomal instability, we developed Stochastically Emergent Tumors (SETs) by inducing mismatch repair deficiency in non-malignant precursors, then engrafting in mice. Barcoded SETs exhibited increased tumoral and drug target heterogeneity over current models. SETs delineated the stochastic contributions, mutational landscapes, and selective pressures distinguishing tumorigenesis from non-malignant precursor in vitro growth. SETs are an unlimited source for diverse Stochastically Emergent cell Lines (SELs), bolstering under-represented cancers. Since SETs composition dynamically reflects therapy exposure, they are a whole-genome platform for precision oncology. We identified three novel genetic drivers (ZFHX3, CIC, KMT2D) of differential prostate cancer therapy responses. These alterations are enriched in patients of African and Chinese ancestry and correlate with significant differences in survival.
]]></description>
<dc:creator>Moussavi-Baygi, R.</dc:creator>
<dc:creator>Ryan, M. J.</dc:creator>
<dc:creator>Sim, W.</dc:creator>
<dc:creator>Hoelscher, S. B.</dc:creator>
<dc:creator>Luga, V.</dc:creator>
<dc:creator>Chandrakumar, A. A.</dc:creator>
<dc:creator>Hoes, L.</dc:creator>
<dc:creator>Cha, J.</dc:creator>
<dc:creator>Lee, Y. S.</dc:creator>
<dc:creator>Herm, K.</dc:creator>
<dc:creator>Doron, B.</dc:creator>
<dc:creator>Ding, C. K. C.</dc:creator>
<dc:creator>Stohr, B. A.</dc:creator>
<dc:creator>Jin, P.</dc:creator>
<dc:creator>Nadkarni, T.</dc:creator>
<dc:creator>Fang, X.</dc:creator>
<dc:creator>Haryono, M.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Karthaus, W. R.</dc:creator>
<dc:creator>Sawyers, C.</dc:creator>
<dc:creator>Feng, F. Y.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Bose, R.</dc:creator>
<dc:date>2025-03-18</dc:date>
<dc:identifier>doi:10.1101/2025.03.17.643814</dc:identifier>
<dc:title><![CDATA[Stochastically Emergent Tumors offer in vivo whole genome interrogation of cancer evolution from non-malignant precursors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.20.644389v1?rss=1">
<title>
<![CDATA[
In vivo validation of the palmitoylation cycle as a therapeutic target in NRAS-mutant cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.20.644389v1?rss=1</link>
<description><![CDATA[
Normal and oncogenic Ras proteins are functionally dependent on one or more lipid modifications1,2. Whereas K-Ras4b farnesylation is sufficient for stable association with the plasma membrane, farnesylated H-Ras, K-Ras4a, and N-Ras traffic to the Golgi where they must undergo palmitoylation before regulated translocation to cell membranes. N-Ras palmitoylation by the DHHC family of palmitoyl acyl transferases (PATs) and depalmitoylation by ABHD17 serine hydrolases is a dynamic process that is essential for the growth of acute myeloid leukemias (AMLs) harboring oncogenic NRAS mutations3-6. Here, we have tested whether co-targeting ABHD17 enzymes and Ras signal output would cooperatively inhibit the proliferation and survival of NRAS-mutant AMLs while sparing normal tissues that retain K-Ras4b function. We show that ABD778, a potent and selective ABHD17 inhibitor with in vivo activity, selectively reduces the growth of NRAS-mutant AML cells in vitro and is synergistic with the allosteric MEK inhibitor PD0325901 (PD901)7,8. Similarly, ABD778 and PD901 significantly extended the survival of recipient mice transplanted with three independent primary mouse AMLs harboring an oncogenic NrasG12Ddriver mutation. Resistant leukemias that emerged during continuous drug treatment acquired by-pass mutations that confer adaptive drug resistance and increase mitogen activated protein kinase (MAPK) signal output. ABD778 augmented the anti-leukemia activity of the pan-PI3 kinase inhibitor pictilisib9, the K/N-RasG12C inhibitor sotorasib10, and the FLT3 inhibitor gilteritinib11. Co-treatment with ABD778 and gilteritinib restored drug sensitivity in a patient-derived xenograft model of adaptive resistance to FLT3 inhibition. These data validate the palmitoylation cycle as a promising therapeutic target in AML and support exploring it in other NRAS-mutant cancers.
]]></description>
<dc:creator>Decker, M.</dc:creator>
<dc:creator>Huang, B. J.</dc:creator>
<dc:creator>Ware, T.</dc:creator>
<dc:creator>Boone, C.</dc:creator>
<dc:creator>Tang, M.</dc:creator>
<dc:creator>Ybarra, J.</dc:creator>
<dc:creator>Ballapuram, A. C.</dc:creator>
<dc:creator>Taran, K. A.</dc:creator>
<dc:creator>Chen, P.-Y.</dc:creator>
<dc:creator>Amendariz, M.</dc:creator>
<dc:creator>Leung, C. J.</dc:creator>
<dc:creator>Harris, M.</dc:creator>
<dc:creator>Tjoa, K.</dc:creator>
<dc:creator>Hongo, H.</dc:creator>
<dc:creator>Abelson, S.</dc:creator>
<dc:creator>Rivera, J.</dc:creator>
<dc:creator>Ngo, N.</dc:creator>
<dc:creator>Herbst, D. M.</dc:creator>
<dc:creator>Suciu, R. M.</dc:creator>
<dc:creator>Guijas, C.</dc:creator>
<dc:creator>Sedighi, K.</dc:creator>
<dc:creator>Andalis, T.</dc:creator>
<dc:creator>Roche, E.</dc:creator>
<dc:creator>Xie, B.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Smith, C. C.</dc:creator>
<dc:creator>Stieglitz, E.</dc:creator>
<dc:creator>Niphakis, M. J.</dc:creator>
<dc:creator>Cravatt, B.</dc:creator>
<dc:creator>Shannon, K.</dc:creator>
<dc:date>2025-03-21</dc:date>
<dc:identifier>doi:10.1101/2025.03.20.644389</dc:identifier>
<dc:title><![CDATA[In vivo validation of the palmitoylation cycle as a therapeutic target in NRAS-mutant cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.21.644645v1?rss=1">
<title>
<![CDATA[
LSD reconfigures the frequency-specific network landscape of the human brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.21.644645v1?rss=1</link>
<description><![CDATA[
Lysergic acid diethylamide (LSD) and other psychedelic substances profoundly alter human consciousness. While several studies have demonstrated changes in brain function and connectivity associated with psychedelics, we still have a limited understanding of how LSD reshapes brain networks operating across different frequency bands. In this study, we applied the recently developed FREQ-NESS method to MEG data from 14 healthy participants who received LSD under four conditions: eyes-closed with or without music and eyes-open with or without a video stimulus. LSD significantly restructures canonical networks in the alpha and beta bands. Relative to broadband brain activity, it enhances the prominence of high alpha (12.1, 13.3 Hz) across all experimental conditions and high beta (25.3 Hz) in three conditions. Conversely, LSD decreases the prominence of low beta (18.1, 19.3 Hz) in both Open and Closed conditions and low alpha (8.5 Hz) in the latter. In addition, LSD substantially alters the spatial distributions or topographies of the networks. Under LSD, the low alpha (8.5 Hz) network shifts anteriorly toward the motor cortex, while high alpha (12.1, 13.3 Hz) becomes more localized to the visual cortex. Low beta (18.1, 19.3 Hz) expands over the temporal and occipital cortices, whereas high beta (25.3, 26.5 Hz) topographies remain unchanged. Our findings provide critical insights into the specific frequencies and spatial networks in which LSD modulates brain connectivity, adding nuance to prevailing theories about network disintegration under psychedelics.

Significance statementPsychedelic substances such as lysergic acid diethylamide (LSD) profoundly alter perception and cognition, yet their effect on brain activity across different frequency bands remains unclear. Using FREQ-NESS applied to MEG data, we show that LSD reorganizes brain networks by enhancing high-frequency alpha and beta rhythms while suppressing lower-frequency counterparts. These changes occur across various experimental conditions and shift the spatial distribution of brain activity, particularly in motor and visual regions. Our findings suggest that LSD modulates brain connectivity in a frequency- and region-specific manner, offering new insights into how psychedelics alter consciousness. This work advances our understanding of LSDs neural effects, which may be relevant for therapeutic applications and models of brain function.
]]></description>
<dc:creator>Shinozuka, K.</dc:creator>
<dc:creator>Rosso, M.</dc:creator>
<dc:creator>Coleman, C. R.</dc:creator>
<dc:creator>Roseman, L.</dc:creator>
<dc:creator>Kaelen, M.</dc:creator>
<dc:creator>Muthukumaraswamy, S. D.</dc:creator>
<dc:creator>Nutt, D. J.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Vuust, P.</dc:creator>
<dc:creator>Kringelbach, M. L.</dc:creator>
<dc:creator>Bonetti, L.</dc:creator>
<dc:date>2025-03-24</dc:date>
<dc:identifier>doi:10.1101/2025.03.21.644645</dc:identifier>
<dc:title><![CDATA[LSD reconfigures the frequency-specific network landscape of the human brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.24.645090v1?rss=1">
<title>
<![CDATA[
Membrane-associated polymerases deliver most of the actin subunits to a lamellipodial network 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.24.645090v1?rss=1</link>
<description><![CDATA[
Actin filaments are two-stranded protein polymers that form the basic structural unit of the eukaryotic actin cytoskeleton. While filaments assembled from purified actin in vitro elongate when soluble monomers bind to free filament ends, in cells the mechanism of filament elongation is less clear. Most monomeric actin in the cytoplasm is bound to the accessory protein profilin, and many regulators of filament assembly recruit actin-profilin complexes to membrane surfaces where they locally accelerate filament elongation. Employing quantitative live-cell imaging of actin-profilin fusion proteins and biochemically defined mutants of the branched actin regulator, WAVE1, we find that only [~]25% of the actin in leading-edge lamellipodial networks enters directly from solution, while the majority enters via membrane-associated polymerases.
]]></description>
<dc:creator>Skruber, K.</dc:creator>
<dc:creator>Sept, D.</dc:creator>
<dc:creator>Mullins, D.</dc:creator>
<dc:date>2025-03-25</dc:date>
<dc:identifier>doi:10.1101/2025.03.24.645090</dc:identifier>
<dc:title><![CDATA[Membrane-associated polymerases deliver most of the actin subunits to a lamellipodial network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.24.645104v1?rss=1">
<title>
<![CDATA[
High-Throughput Screening on Primary Tumor-Associated Microglia and Macrophages Identifies HDAC Inhibitors as Enhancers of Phagocytosis and Potent Partners for Immunotherapy in Glioblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.24.645104v1?rss=1</link>
<description><![CDATA[
Glioblastoma multiforme (GBM) is a lethal brain tumor with limited treatment options. Tumor-associated macrophages and microglia (TAMs) drive immune suppression and tumor progression, making them a key therapeutic target for GBM. Enhancing TAM phagocytosis in GBM has shown promise, particularly with innate checkpoint inhibitors, such as CD47-blocking antibodies. However, small molecule approaches, which offer tunable and potentially synergistic mechanisms, remain underexplored in this context. In this study, we conducted the first large-scale chemical screen on primary TAMs from patients with GBM, identifying histone deacetylase (HDAC) inhibitors as potent inducers of phagocytosis. These compounds demonstrated phagocytosis-inducing effects across multiple GBM patient samples, with further amplification when combined with CD47 blockade. In a xenograft GBM model, HDAC inhibitors enhanced phagocytosis and suppressed tumor growth, with even greater efficacy in combination with CD47 antibodies. Our findings highlight HDAC inhibitors as promising agents to reprogram TAMs and synergize with immune checkpoint therapies, offering a novel strategy to bolster anti-tumor immunity in GBM.
]]></description>
<dc:creator>Khalaj, M.</dc:creator>
<dc:creator>Gutierrez, M. L.</dc:creator>
<dc:creator>Nejad, P.</dc:creator>
<dc:creator>Raveh, T.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:creator>Weissman, I. L.</dc:creator>
<dc:date>2025-03-27</dc:date>
<dc:identifier>doi:10.1101/2025.03.24.645104</dc:identifier>
<dc:title><![CDATA[High-Throughput Screening on Primary Tumor-Associated Microglia and Macrophages Identifies HDAC Inhibitors as Enhancers of Phagocytosis and Potent Partners for Immunotherapy in Glioblastoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.26.645557v1?rss=1">
<title>
<![CDATA[
A multi-modal cell-free RNA language model for liquid biopsy applications 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.26.645557v1?rss=1</link>
<description><![CDATA[
Cell-free RNA (cfRNA) profiling has emerged as a powerful tool for non-invasive disease detection, but its application is limited by data sparsity and complexity, especially in settings with constrained sample availability. We introduce Exai-1, a multi-modal, transformer-based generative foundation model that integrates RNA sequence embeddings with cfRNA abundance data to capture biologically meaningful representations of circulating RNAs. By leveraging both sequence and expression modalities, Exai-1 captures a biologically meaningful latent structure of cfRNA profiles. Pre-trained on over 306 billion tokens from 8,339 samples, Exai-1 enhances signal fidelity, reduces technical noise, and improves disease detection by generating synthetic cfRNA profiles. We show that self-attention and variational inference are particularly important for preservation of biological signals and contextual relationships. Additionally, Exai-1 facilitates cross-biofluid translation and assay compatibility through disentangling biological signals from confounders. By uniting sequence-informed embeddings with cfRNA expression patterns, Exai-1 establishes a transfer learning foundation for liquid biopsy, offering a scalable and adaptable framework for next-generation cfRNA-based diagnostics.
]]></description>
<dc:creator>Karimzadeh, M.</dc:creator>
<dc:creator>Sababi, A. M.</dc:creator>
<dc:creator>Momen-Roknabadi, A.</dc:creator>
<dc:creator>Chen, N.-C.</dc:creator>
<dc:creator>Cavazos, T. B.</dc:creator>
<dc:creator>Sekhon, S.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Hanna, R.</dc:creator>
<dc:creator>Huang, A.</dc:creator>
<dc:creator>Nguyen, D.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Lam, T.</dc:creator>
<dc:creator>Hartwig, A.</dc:creator>
<dc:creator>Fish, L.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Behsaz, B.</dc:creator>
<dc:creator>Hormozdiari, F.</dc:creator>
<dc:creator>Alipanahi, B.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2025-04-01</dc:date>
<dc:identifier>doi:10.1101/2025.03.26.645557</dc:identifier>
<dc:title><![CDATA[A multi-modal cell-free RNA language model for liquid biopsy applications]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.01.646243v1?rss=1">
<title>
<![CDATA[
Smoothened inhibition of PKA at cilia transduces Hedgehog signals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.01.646243v1?rss=1</link>
<description><![CDATA[
Hedgehog (HH) signaling in vertebrates is dependent on the primary cilium, an organelle that scaffolds signal transduction. HH signals induce Smoothened (SMO) enrichment in the cilium and indirectly triggers the conversion of GLI proteins into transcriptional activators of HH target genes. Recently, SMO has been shown to inhibit protein kinase A (PKA). To test the hypothesis that SMO specifically inhibits PKA at cilia to activate the HH signal transduction pathway, we developed a ciliary PKA biosensor. Activation of the HH signal transduction pathway by either Sonic hedgehog (SHH) or SMO agonist (SAG) inhibited ciliary PKA activity. Blocking SMO phosphorylation by GRK2/3 prevented ciliary SMO from inhibiting ciliary PKA activity. Gi was dispensable for SMO inhibition of ciliary PKA. In contrast, mutating the SMO C-terminal tail protein kinase inhibitor (PKI) pseudosubstrate site interfered with the ability of SMO to inhibit ciliary PKA. Therefore, HH signaling is transduced via SMO direct inhibition of PKA at cilia, in a manner dependent on GRK2/3.
]]></description>
<dc:creator>Nguyen, T. D.</dc:creator>
<dc:creator>Konjikusic, M. J.</dc:creator>
<dc:creator>Castillo, L. D.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:date>2025-04-01</dc:date>
<dc:identifier>doi:10.1101/2025.04.01.646243</dc:identifier>
<dc:title><![CDATA[Smoothened inhibition of PKA at cilia transduces Hedgehog signals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.02.646699v1?rss=1">
<title>
<![CDATA[
Effect of Azithromycin treatment on the microbial composition, functional dynamics and resistomes of endocervical, vaginal and rectal microbiomes of women in Fiji with Chlamydia trachomatis infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.02.646699v1?rss=1</link>
<description><![CDATA[
Antibiotics disrupt mucosal microbial communities, yet the effects on microbiomes with Chlamydia trachomatis (Ct) infection remain poorly understood. Some data exist on vaginal microbiomes pre- and post-treatment, but none are available for the endocervix or rectum that are primary sites of infection. We applied metagenomic shotgun sequencing to vaginal, endocervical and rectal samples from women who, overtime, had Ct persistence, clearance, or no infection to evaluate azithromycin-induced changes in microbial composition, function, and the resistome. Our results show a shift in composition and function that support Ct post-treatment with azithromycin resistance mutations in the Ct rplV gene and significant endocervical enrichment of azithromycin resistance genes in Lactobacillus iners and Gardnerella vaginalis, the strains of which have moderate/high potential for biofilm formation. These findings highlight the unintended ecological consequences of azithromycin, including resistance gene propagation and microbiome shifts that support persistent/recurrent Ct, emphasizing the need for novel treatment and microbiome- preserving strategies.
]]></description>
<dc:creator>Bommana, S.</dc:creator>
<dc:creator>Olagoke, S.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Kama, M.</dc:creator>
<dc:creator>Dehdashti, M.</dc:creator>
<dc:creator>Kodimerla, R.</dc:creator>
<dc:creator>Read, T.</dc:creator>
<dc:creator>Dean, D.</dc:creator>
<dc:date>2025-04-02</dc:date>
<dc:identifier>doi:10.1101/2025.04.02.646699</dc:identifier>
<dc:title><![CDATA[Effect of Azithromycin treatment on the microbial composition, functional dynamics and resistomes of endocervical, vaginal and rectal microbiomes of women in Fiji with Chlamydia trachomatis infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.01.646034v1?rss=1">
<title>
<![CDATA[
Improving interpretability of transcription factor binding models with DNA shape features 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.01.646034v1?rss=1</link>
<description><![CDATA[
Deep learning models in genomics that predict molecular phenotypes from DNA sequence traditionally focus on one-hot encoded nucleotide representations. Here, we develop a novel model that extends this approach by incorporating DNA structural attributes indicative of local DNA shape alongside canonical sequence inputs. This augmentation provides an additional axis for model interpretability and aids in identifying regulatory patterns not apparent from sequence alone. Applying this approach to prediction of transcription factor binding (ChIP-seq) demonstrates that combining sequence and structural DNA information can improve the identification of regulatory elements to provide a more nuanced understanding of genomic function and regulation.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=96 SRC="FIGDIR/small/646034v1_ufig1.gif" ALT="Figure 1">
View larger version (23K):
org.highwire.dtl.DTLVardef@b648b6org.highwire.dtl.DTLVardef@15fa931org.highwire.dtl.DTLVardef@15d0d31org.highwire.dtl.DTLVardef@d212e_HPS_FORMAT_FIGEXP  M_FIG C_FIG Schematic overview of the DeepShape model. One-hot encoded sequence, and five DNA shape attributes--minor groove width (MGW), helical twist (HelT), propeller twist (ProT), roll, and electrostatic potential (EP)--are input separately to sequence and shape branches of the model, each processed through two convolutional layers and subsequently concatenated for further processing through additional convolutional layers.
]]></description>
<dc:creator>Keivanfar, R. L.</dc:creator>
<dc:creator>Yang, F.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Ioannidis, N. M.</dc:creator>
<dc:date>2025-04-03</dc:date>
<dc:identifier>doi:10.1101/2025.04.01.646034</dc:identifier>
<dc:title><![CDATA[Improving interpretability of transcription factor binding models with DNA shape features]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.08.647866v1?rss=1">
<title>
<![CDATA[
Autism-associated Scn2a haploinsufficiency disrupts in vivo dendritic signaling and impairs flexible decision-making 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.08.647866v1?rss=1</link>
<description><![CDATA[
SCN2A is a high-confidence risk gene for autism spectrum disorder. Loss-of-function mutations in Scn2a reduce dendritic excitability in neocortical pyramidal cells. However, the impact of Scn2a haploinsufficiency on dendritic signaling in vivo, particularly during behavior, is unknown. In this study, we used two-photon microscopy to image dendritic calcium transients in deep layer pyramidal cells in the mouse medial frontal cortex. Scn2a+/- mice had diminished coupling between apical and proximal dendritic compartments. Pyramidal tract neurons had abnormal event rates, while intratelencephalic neurons had compartment-specific alterations indicative of diminished dendritic integration. In a matching pennies task, Scn2a+/- mice were inflexible in the face of changing competitive pressure. Apical dendritic tuft in IT neurons typically encoded reward and strategy, but these task-specific representations were altered in Scn2a+/- mice. Collectively, the findings demonstrate that Scn2a haploinsufficiency weakens dendritic integration in vivo and disrupts the dendritic encoding of decision variables, potentially contributing to the cognitive rigidity in autism spectrum disorder.
]]></description>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Shen, L.</dc:creator>
<dc:creator>Indajang, J.</dc:creator>
<dc:creator>Savalia, N. K.</dc:creator>
<dc:creator>Johnson, T. G.</dc:creator>
<dc:creator>Qu, J.</dc:creator>
<dc:creator>Bender, K. J.</dc:creator>
<dc:creator>Kwan, A. C.</dc:creator>
<dc:date>2025-04-09</dc:date>
<dc:identifier>doi:10.1101/2025.04.08.647866</dc:identifier>
<dc:title><![CDATA[Autism-associated Scn2a haploinsufficiency disrupts in vivo dendritic signaling and impairs flexible decision-making]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.13.648587v1?rss=1">
<title>
<![CDATA[
Telomere Orchestrates Encapsulation of Mitotic Chromosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.13.648587v1?rss=1</link>
<description><![CDATA[
Telomere is crucial for continued cell proliferation and survival. Using high-temporal microscopy, we show that in mitosis it provides an initiation site for reassembly of barrier/scaffolding proteins during nuclear envelop reassembly. Subsequent spread of these proteins onto decondensing chromosomes completes encapsulation of the entire chromosome set. Depletion of telomere-tip components, TRF1 and TRF2, randomizes nuclear envelope reassembly initiation sites leading to multiple micronuclei production. The data show that telomere tip orchestrates this precisely timed program of nuclear envelop re-assembly.

One Sentence SummaryYang and Mikawa investigated the role of telomere in initiating and forming a single nucleus.
]]></description>
<dc:creator>Yang, P. B.</dc:creator>
<dc:creator>Mikawa, T.</dc:creator>
<dc:date>2025-04-14</dc:date>
<dc:identifier>doi:10.1101/2025.04.13.648587</dc:identifier>
<dc:title><![CDATA[Telomere Orchestrates Encapsulation of Mitotic Chromosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.10.648051v1?rss=1">
<title>
<![CDATA[
Submicron-Sampling of Living Cells by Macrophages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.10.648051v1?rss=1</link>
<description><![CDATA[
SUMMARY PARAGRAPHAn effective immune system must sample and appreciate healthy-self identity to prevent autoimmunity and to contrast to pathogenic insults1-3. Self-proteins are presented to T cells in the thymus during immune cell development2,3, and must be presented throughout the body to both maintain regulatory T cell populations4-6 and provide a tonic signal to maintain conventional T cells over time7-9. The ready observations of continuous apoptosis in some organs together with the ingestion of that material by myeloid populations has led to a conventional understanding of ongoing cell-death as a major source of self-antigens10, complemented in some situations by uptake of free-floating cell-derived vesicles. Here, we used a series of companion imaging and vesicular labeling technologies to reveal an alternate process undertaken by macrophages that results in non-destructive and direct sampling of living cells. The process requires cell-cell contact, does not require caspase activation, and takes place via a trogocytosis-like stretching of the target cell into the macrophage, leading to the generation of submicron-sized vesicles containing cytoplasm. Using a high-dimensional flow-based method for labeling vesicles ingested under this versus other conditions, we find that live-sampled material is distinctly processed, is poorly subject to fusion with lysosomes, and produces ensuing differential effects on the presentation of those to CD4 versus CD8 T cells. Disrupting this trafficking by redirecting antigen to the lysosome significantly reduced the associated macrophage-mediated priming of CD8 T cells. This demonstrates an important and substantial sampling of living cells by the immune system, with clear consequences for maintaining the border of immunity.
]]></description>
<dc:creator>Fan, A. C.</dc:creator>
<dc:creator>Thota, R.</dc:creator>
<dc:creator>Serwas, N.</dc:creator>
<dc:creator>Marchuk, K.</dc:creator>
<dc:creator>Ruhland, M.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2025-04-16</dc:date>
<dc:identifier>doi:10.1101/2025.04.10.648051</dc:identifier>
<dc:title><![CDATA[Submicron-Sampling of Living Cells by Macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.15.649018v1?rss=1">
<title>
<![CDATA[
Archaeal SegAB forms a bipolar structure that promotes chromosome segregation in spherical cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.15.649018v1?rss=1</link>
<description><![CDATA[
Archaeal segAB operons are thought to promote chromosome segregation, but their mechanism remains unknown. We employ comparative genomics, structural biology, genetic knockouts, and quantitative cell biology to investigate how SegA and SegB proteins work together to segregate chromosomes in the thermophilic archaeon Sulfolobus acidocaldarius. In vitro, SegB binds a centromeric DNA sequence adjacent to the segAB operon, and in vivo forms a distinct focus on each segregating chromosome. SegA, a ParA-like ATPase, binds DNA non-specifically in vitro and promotes chromosome compaction and segregation in vivo. During division, SegA shifts from chromosome-associated puncta to form a single, elongated figure that runs between separating SegB foci. Late in division, SegA retreats to regions surrounding separated SegB foci. Elongated SegA figures appear in segB knockout cells but no longer lie perpendicular to the division plane. We propose that SegA and SegB interact to form a bipolar, DNA-segregating structure radically different from bacterial ParABS systems.
]]></description>
<dc:creator>Charles-Orszag, A.</dc:creator>
<dc:creator>Lord, S. J.</dc:creator>
<dc:creator>Herrera, N.</dc:creator>
<dc:creator>Strauskulage, L.</dc:creator>
<dc:creator>Bhowmick, A.</dc:creator>
<dc:creator>Goddard, T.</dc:creator>
<dc:creator>Wassmer, B.</dc:creator>
<dc:creator>van Wolferen, M.</dc:creator>
<dc:creator>Asper, G.</dc:creator>
<dc:creator>Flis, A.</dc:creator>
<dc:creator>Rodriguez, J.</dc:creator>
<dc:creator>Redding, S.</dc:creator>
<dc:creator>Rosenberg, O.</dc:creator>
<dc:creator>Albers, S.-V.</dc:creator>
<dc:creator>Mullins, D.</dc:creator>
<dc:date>2025-04-16</dc:date>
<dc:identifier>doi:10.1101/2025.04.15.649018</dc:identifier>
<dc:title><![CDATA[Archaeal SegAB forms a bipolar structure that promotes chromosome segregation in spherical cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.11.648468v1?rss=1">
<title>
<![CDATA[
Dissociating physiological ripples and epileptiform discharges with vision transformers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.11.648468v1?rss=1</link>
<description><![CDATA[
Two frequently studied bursts of neural activity in the hippocampus are normal physiological ripples and abnormal interictal epileptiform discharges (IEDs). While they are different waveforms, IEDs are notoriously picked up as false positives when using typical automated ripples detectors which are prone to sharp edge artifacts. This has created challenges for studying ripples and IEDs independently. We leveraged recent advances in computer vision on time-frequency feature representations to enable more comprehensive and objective dissociation of these phenomena. We retrospectively evaluated human intracranial recordings from 46 hippocampal depth electrode sites among 17 patients with focal epilepsy, the majority of whom had a seizure-onset zone/network involving the hippocampus. We implemented a common human ripple detection algorithm and broadband spectrograms of all detected "ripple candidates" were projected into low-dimensional space. We segmented them using k-means to infer pseudo-labels for probable ripples and probable IEDs. Independently, human expert IED labels were manually annotated for comparison. State-of-the-art vision transformer models were implemented on individual spectrograms to approach ripple vs. IED dissociation as an image classification problem. We detected 31,847 ripple/IED candidates, and a median 3.9% per patient (range: 0-47.2%) were IEDs based on expert label overlap. Low-dimensional projection of spectrograms separated canonical IEDs vs. ripples better than raw or ripple-filtered waveforms. Canonical ripple and IED candidates emerged at opposite poles with a continuous landscape of intermediates in between. A binary vision transformer model trained on expert-labeled IED vs. non-IED candidate spectrograms with 5-fold cross-validation showed a mean area under the curve (AUC) of 0.970 and mean precision-recall curve of 0.694, both significantly above chance. To evaluate generalizability, we implemented a leave-one-patient-out cross-validation approach, in which training on pseudo-labels and testing on expert-labeled data demonstrated near-expert performance (mean AUC 0.966 across patients, range 0.892-0.997). Transformer-derived attention maps revealed that models were tuned to triangle-like edge artifact spatial features in the spectrograms. Model-derived probabilities (i.e. of being an IED) for all candidates demonstrated continuous transitions between ripples vs. IEDs, as opposed to binary clustering. The delineation between ripples and IEDs appears best represented as a gradient (i.e. not binary) due to physiological ripple features overlapping with sharpened and/or high frequency pathophysiological IED features. Vision transformers nevertheless perform virtually at human expert levels in dissociating these phenomena by leveraging time-frequency spatial features enabled by neural data spectrograms. Such tools applied to spectrotemporal representations may augment comprehensive investigations in cognitive neurophysiology and epileptiform signal biomarker optimization for closed-loop applications.
]]></description>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Kleen, J.</dc:creator>
<dc:date>2025-04-18</dc:date>
<dc:identifier>doi:10.1101/2025.04.11.648468</dc:identifier>
<dc:title><![CDATA[Dissociating physiological ripples and epileptiform discharges with vision transformers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.15.648984v1?rss=1">
<title>
<![CDATA[
β-III tubulin identifies anti-fibrotic state of pericytes in pulmonary fibrosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.15.648984v1?rss=1</link>
<description><![CDATA[
Pericytes have been implicated in pulmonary fibrosis, yet their activated cellular states and functional roles remain largely unclear. Here, we identified {beta}-III tubulin (Tuj1) as a distinctive marker for fibrosis-associated lung pericytes. Most pericytes in fibrotic regions are Tuj1-positive and interact uniquely with multiple endothelial cells, localizing near collagen-producing fibroblasts and pro-fibrotic SPP1+/arginase+ macrophages. Tuj1 expression is predominantly induced in pericytes during the fibrotic phase, and Tuj1 gene (Tubb3) knockout in mice exacerbates lung fibrosis, accompanied by an increase in the neighboring pro-fibrotic fibroblasts and macrophages, suggesting an anti-fibrotic role for Tuj1-expressing pericytes. Mechanistically, the anti-fibrotic chemokine CXCL10 is upregulated in Tuj1-expressing pericytes, whereas this upregulation is not observed in Tubb3 knockout. Moreover, CXCL10 inhibits the pro-fibrotic differentiation of macrophages induced by lung fibroblasts in culture, implying that CXCL10 may mediate the anti-fibrotic effects of Tuj1-expressing pericytes. These findings underscore the role of lung pericytes in negatively regulating fibrotic process and their potential as therapeutic targets for pulmonary fibrosis patients.

SummaryIn response to fibrotic stimuli, lung pericytes upregulate {beta}-III tubulin (Tuj1) expression, adopting an anti-fibrotic phenotype. This phenotype acts as a negative immune regulator of pro-fibrotic macrophage differentiation by releasing CXCL10, thereby counteracting the progression of pulmonary fibrosis.
]]></description>
<dc:creator>Sato, R.</dc:creator>
<dc:creator>Imamura, K.</dc:creator>
<dc:creator>Tsukui, T.</dc:creator>
<dc:creator>Yoshida, T.</dc:creator>
<dc:creator>Tomita, Y.</dc:creator>
<dc:creator>Fujino, K.</dc:creator>
<dc:creator>Ikeda, T.</dc:creator>
<dc:creator>Onizawa, K.</dc:creator>
<dc:creator>Sogo, T.</dc:creator>
<dc:creator>Combs, C. A.</dc:creator>
<dc:creator>Murgai, M.</dc:creator>
<dc:creator>Kopp, J. B.</dc:creator>
<dc:creator>Suzuki, M.</dc:creator>
<dc:creator>Sakagami, T.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:creator>Mukouyama, Y.-s.</dc:creator>
<dc:date>2025-04-19</dc:date>
<dc:identifier>doi:10.1101/2025.04.15.648984</dc:identifier>
<dc:title><![CDATA[β-III tubulin identifies anti-fibrotic state of pericytes in pulmonary fibrosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.15.649017v1?rss=1">
<title>
<![CDATA[
Survey of hippocampal responses to sound in naive mice reveals widespread activation by broadband noise onsets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.15.649017v1?rss=1</link>
<description><![CDATA[
Recent studies suggest some hippocampal (HC) neurons respond to passively presented sounds in naive subjects, but the specificity and prevalence of these responses remain unclear. We used Neuropixels probes to record unit activity in HC and auditory cortex (ACtx) of awake, untrained mice during presentation of diverse sound stimuli. A subset of HC neurons exhibited reliable, short-latency responses to passive sounds, including tones and broadband noise. HC units showed evidence of tuning for tone frequency but not spectrotemporal features in continuous dynamic moving ripples. Across sound types, HC responses overwhelmingly occurred at stimulus onset; they quickly adapted to continuous sounds and did not respond at sound offset. Among all sounds tested, broadband noise was by far most effective at driving HC activity, with response prevalence scaling with increasing spectral bandwidth and density. Responses to noise were also far more common than visual flash stimuli. Sound-evoked face movements, quantified by total facial motion energy (FME), correlated with population-level HC activity, but many individual units responded regardless of movement, indicating both auditory and motor-related inputs. These results show that abrupt, acoustic events are sufficient to activate HC neurons in the absence of learning or behavioral engagement. This suggests a possible role for HC in detecting salient environmental changes and supports the idea that auditory inputs contribute directly to HC function. Given emerging links between hearing loss and dementia, these findings highlight a potential pathway by which auditory deafferentation could impact cognitive health.
]]></description>
<dc:creator>Bigelow, J.</dc:creator>
<dc:creator>Suzuki, T.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Hasenstaub, A. R.</dc:creator>
<dc:date>2025-04-20</dc:date>
<dc:identifier>doi:10.1101/2025.04.15.649017</dc:identifier>
<dc:title><![CDATA[Survey of hippocampal responses to sound in naive mice reveals widespread activation by broadband noise onsets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.15.649026v1?rss=1">
<title>
<![CDATA[
Improved Injury Detection Through Harmonizing Multi-Site Neuroimaging Data after Experimental TBI: A Translational Outcomes Project in NeuroTrauma (TOP-NT) Consortium Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.15.649026v1?rss=1</link>
<description><![CDATA[
Multi-site neuroimaging studies have become increasingly common in order to generate larger samples of reproducible data to answer questions associated with smaller effect sizes. The data harmonization model NeuroCombat has been shown to remove site effects introduced by differences in site-related technical variance while maintaining group differences, yet its effect on improving statistical power in pre-clinical models of CNS disease is unclear. The present study examined fractional anisotropy data computed from diffusion weighted imaging data at 3 and 30 days post-controlled cortical impact injury from 184 adult rats across four sites as part of the Translational-Outcome-Project-in-Neurotrauma (TOP-NT) Consortium. Findings confirmed prior clinical reports that NeuroCombat fails to remove site effects in data containing a high proportion-of-outliers (>5%) and skewness, which introduced significant variation in non-outlier sites. After removal of one outlier site and harmonization using a global sham population, harmonization displayed an increase in effect size in data that displayed group level effects (p<0.01) in both univariate and voxel-level volumes of pathology. This was characterized by movement toward similar distributions in voxel measurements (Kolmogorov-Smirnov p<<0.001 to >0.01) and statistical power increases within the ipsilateral cortex. Harmonization improved statistical power and frequency of significant differences in areas with existing group differences, thus improving the ability to detect regions affected by injury rather than by other confounds. These findings indicate the utility of NeuroCombat in reproducible data collection, where biological differences can be accurately revealed to allow for greater reliability in multi-site neuroimaging studies.

Significance StatementThis project demonstrates the utility of NeuroCombat in reducing site effects in multi-site rodent imaging. We also demonstrate that harmonization improves the ability to distinguish between sham and injured rats at the voxel level and increase statistical power and effect size in areas of injury. Multi-center studies are becoming more common to allow for increased efficiency in data collection, and with conservative approaches and analysis into the datasets, NeuroCombat can be utilized to improve study reliability and reproducibility.
]]></description>
<dc:creator>Kislik, G.</dc:creator>
<dc:creator>Fox, R.</dc:creator>
<dc:creator>Korotcov, A. V.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Febo, M.</dc:creator>
<dc:creator>Moghadas, B.</dc:creator>
<dc:creator>Bibic, A.</dc:creator>
<dc:creator>Zou, Y.</dc:creator>
<dc:creator>Wan, J.</dc:creator>
<dc:creator>Koehler, R. C.</dc:creator>
<dc:creator>Adebayo, T.</dc:creator>
<dc:creator>Burns, M. P.</dc:creator>
<dc:creator>McCabe, J. T.</dc:creator>
<dc:creator>Wang, K. K.</dc:creator>
<dc:creator>Huie, J. R.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Harris, N. G.</dc:creator>
<dc:creator>The TOP-NT Investigators,</dc:creator>
<dc:date>2025-04-20</dc:date>
<dc:identifier>doi:10.1101/2025.04.15.649026</dc:identifier>
<dc:title><![CDATA[Improved Injury Detection Through Harmonizing Multi-Site Neuroimaging Data after Experimental TBI: A Translational Outcomes Project in NeuroTrauma (TOP-NT) Consortium Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.15.649037v1?rss=1">
<title>
<![CDATA[
D3 dopamine receptors implicate a subtype of medium spiny neuron in the aversive effects of antipsychotic medications 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.15.649037v1?rss=1</link>
<description><![CDATA[
Second generation antipsychotics (SGAs) are widely used clinical tools, yet they often cause negative side effects and take weeks to become effective, leading to poor patient compliance. The effect/side effect profile of individual SGAs is highly variable, and the mechanisms that underlie this variability are not well understood. We found that SGA activity at D3 dopamine receptors (D3R) in the Nucleus Accumbens (NAc) mediates the aversive effects of SGAs. Using single-nucleus RNA sequencing, we found that D3R is expressed in a subpopulation of D1R neurons and defines its own distinct NAc cell type. We demonstrate that while multiple SGAs (clozapine and quetiapine) cause acute conditioned place aversion in mice, only chronic treatment with quetiapine, an arrestin-biased agonist at D3R, causes aversion to abate. We further show at both the cell and population level that quetiapine inhibits D3R-neurons in the lateral shell (LatSh) of the NAc. Selective optogenetic inhibition of D3R-neurons in the LatSh produces real time place aversion in mice, implicating this cell type in the aversive effects of SGAs. Our findings suggest a cellular and systems-level mechanism underlying aversion to SGAs and highlight the pathway to selective tolerance to this aversion, providing a framework for future therapeutic strategies in SGA development.
]]></description>
<dc:creator>Lewis, E.</dc:creator>
<dc:creator>Muir, J.</dc:creator>
<dc:creator>Li, L. C.</dc:creator>
<dc:creator>Glienke, J.</dc:creator>
<dc:creator>Gooding, S. W.</dc:creator>
<dc:creator>Bender, K.</dc:creator>
<dc:creator>Kim, C.</dc:creator>
<dc:creator>Whistler, J.</dc:creator>
<dc:date>2025-04-20</dc:date>
<dc:identifier>doi:10.1101/2025.04.15.649037</dc:identifier>
<dc:title><![CDATA[D3 dopamine receptors implicate a subtype of medium spiny neuron in the aversive effects of antipsychotic medications]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.18.649514v1?rss=1">
<title>
<![CDATA[
Essential Role of Protein Kinase R in the Pathogenesis of Pulmonary Veno-occlusive Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.18.649514v1?rss=1</link>
<description><![CDATA[
Pulmonary veno-occlusive disease (PVOD) is a rare and severe subtype of pulmonary arterial hypertension, marked by progressive remodeling of small pulmonary arteries and veins with no therapies. Using a mitomycin C (MMC)-induced rat model, we previously demonstrated that protein kinase R (PKR)-mediated integrated stress response (ISR) drives endothelial dysfunction and vascular remodeling. To determine if PKR is the sole mediator of ISR and the pathogenesis, we treated control (Ctrl) and PKR knockout (KO) mice with the same dose of MMC. Consistent with rat data, Ctrl mice displayed ISR activation, vascular remodeling, and pulmonary hypertension after MMC treatment, while KO mice showed none of these phenotypes. Proteomic analysis revealed that MMC-mediated ISR activation attenuates protein synthesis in Ctrl but not in KO mice. These findings underscore the significance of PKR-dependent ISR activation and subsequent perturbation of proteostasis as central mechanisms driving PVOD pathogenesis and identifying PKR as a promising therapeutic target.
]]></description>
<dc:creator>Prabhakar, A.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Wadhwa, M.</dc:creator>
<dc:creator>Barpanda, A.</dc:creator>
<dc:creator>Lyons, J.</dc:creator>
<dc:creator>Gowda, A.</dc:creator>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Arvind, A.</dc:creator>
<dc:creator>Ghatpande, P.</dc:creator>
<dc:creator>Wiita, A.</dc:creator>
<dc:creator>Graham, B. B.</dc:creator>
<dc:creator>Lagna, G.</dc:creator>
<dc:creator>Hata, A.</dc:creator>
<dc:date>2025-04-23</dc:date>
<dc:identifier>doi:10.1101/2025.04.18.649514</dc:identifier>
<dc:title><![CDATA[Essential Role of Protein Kinase R in the Pathogenesis of Pulmonary Veno-occlusive Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.20.649464v1?rss=1">
<title>
<![CDATA[
Plasma lipidome dysregulation in frontotemporal dementia reveals shared, genotype-specific, and severity-linked alterations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.20.649464v1?rss=1</link>
<description><![CDATA[
Developing new treatment strategies for frontotemporal dementia (FTD) and other forms of neurodegeneration requires biomarkers to monitor disease progression. Dysregulated brain lipid metabolism, particularly sphingolipids enriched in the nervous system, is a key feature of neurodegeneration. However, plasma lipids have been investigated less for their potential as biomarkers than brain imaging and serum proteins. Here we examined the plasma lipidomes of a cohort of heterozygous carriers of gene variants associated with autosomal dominant familial FTD (including GRN, C9orf72, and MAPT loci), comparing them with aged-matched controls. In general, FTD subjects exhibited increases in plasma levels of specific species of gangliosides (GM3(d18:1_16:0), GM3(d18:1_18:0), and GM3(d18:1_24:1)) and ceramide (Cer(d18:1_23:0)) and selected polyunsaturated triacylglycerols (TG). Other species of ceramides (Cer(d18:0_22:0)), phosphatidylethanolamine (PE(18:0_24:0)), and sphingomyelin (SM(38:0)) were reduced in plasma of FTD subjects. Levels of glucosylsphingosine (GlcSph(d18:1)) were elevated specifically in GRN carriers, SM(34:1) was reduced in C9orf72 carriers, and TG(16:0_18:1_20:3)) were decreased in MAPT variant carriers. Notably, the ganglioside GM3(d18:1_16:0) was consistently elevated across all FTD genetic subtypes. Furthermore, the levels of these lipids correlated with disease severity in FTD patients. Our findings suggest that specific plasma lipid changes, notably several sphingolipids, may be useful biomarkers for FTD disease or progression.
]]></description>
<dc:creator>Ambaw, Y. A.</dc:creator>
<dc:creator>Ljubenkov, P. A.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Hamed, A.</dc:creator>
<dc:creator>Boland, S.</dc:creator>
<dc:creator>Boxer, A. L.</dc:creator>
<dc:creator>Walther, T. C.</dc:creator>
<dc:creator>Farese, R.</dc:creator>
<dc:date>2025-04-24</dc:date>
<dc:identifier>doi:10.1101/2025.04.20.649464</dc:identifier>
<dc:title><![CDATA[Plasma lipidome dysregulation in frontotemporal dementia reveals shared, genotype-specific, and severity-linked alterations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.22.649486v1?rss=1">
<title>
<![CDATA[
Spatiotemporal Analysis of Remyelination Reveals a Concerted Interferon-Responsive Glial State That Coordinates Immune Infiltration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.22.649486v1?rss=1</link>
<description><![CDATA[
Remyelination, the process by which axons are re-encased in myelin after injury, is a critical step in restoring brain function, yet the dynamics from initial injury to repair remain poorly characterized. Here, we combined optimized single-nucleus RNA-seq with Slide-seqv2, a high-resolution spatial transcriptomics technology, to densely reconstruct the cellular processes that coordinate remyelination after a focal demyelinating injury. This revealed several findings: First, we found extensive transcriptional diversity of glia and monocyte-derived macrophages from demyelination to repair. Second, we identified a population of infiltrating peripheral lymphocytes--predominantly CD8 T-cells and natural killer cells--that are enriched specifically during remyelination. Third, we identified a concerted interferon-response gene signature that is shared across several cell types--microglia, astrocytes, and the oligodendrocyte lineage--just prior to reestablishment of myelin. These interferon-responsive glia (IRG) form clusters around remyelinating white matter and their formation is solely dependent on the type I interferon receptor. Functionally, we found that IRG secrete the cytokine CXCL10 which mediates infiltration of peripheral lymphocytes into the repairing white matter. Depletion of the most abundant infiltrating lymphocyte, CD8 T-cells, attenuated the differentiation of mature oligodendrocytes during remyelination. Together, our data reveals the diversity of glial-immune interactions that orchestrate white matter repair and a type I-dependent glial state that drives lymphocyte influx into damaged white matter to modulate oligodendrocyte differentiation.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=57 SRC="FIGDIR/small/649486v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Dolan, M.-J.</dc:creator>
<dc:creator>Rakosi-Schmidt, R.</dc:creator>
<dc:creator>Garcia, E.</dc:creator>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Nadaf, N. M.</dc:creator>
<dc:creator>Dixon, J.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:creator>Morshed, N.</dc:creator>
<dc:creator>Depp, C.</dc:creator>
<dc:creator>Doman, J.</dc:creator>
<dc:creator>Xia, J.</dc:creator>
<dc:creator>Jäntti, H.</dc:creator>
<dc:creator>Stevens, B.</dc:creator>
<dc:creator>Macosko, E. Z.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.22.649486</dc:identifier>
<dc:title><![CDATA[Spatiotemporal Analysis of Remyelination Reveals a Concerted Interferon-Responsive Glial State That Coordinates Immune Infiltration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.22.649607v1?rss=1">
<title>
<![CDATA[
Towards individualized deep brain stimulation: A stereoencephalography-based workflow for unbiased neurostimulation target identification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.22.649607v1?rss=1</link>
<description><![CDATA[
ObjectivesDeep brain stimulation (DBS) is increasingly being used to treat a variety of neuropsychiatric conditions, many of which exhibit idiosyncratic symptom presentations and neural correlates across individuals. Thus, we have utilized inpatient stereoelectroencephalography (sEEG) to identify personalized therapeutic stimulation sites for chronic implantation of DBS. Informed by our experience, we have developed a statistics-driven framework for stimulation testing to identify therapeutic targets.

Materials and MethodsFourteen participants (major depressive disorder = 6, chronic pain = 6, obsessive-compulsive disorder = 2) underwent inpatient testing using sEEG and symptom monitoring to identify personalized stimulation targets for subsequent DBS implantation. We present a structured approach to this sEEG testing, integrating a Stimulation Testing Decision Tree with power analysis and effect size considerations to inform adequately powered results to detect therapeutic stimulation sites with statistical rigor.

ResultsEffect sizes (Hedges g) of stimulation-induced symptom score changes ranged from -1.5 to +2.39. The standard deviation of sham trial responses was a strong predictor of stimulation response variability, as confirmed by a leave-one-out cross-validated linear regression (R2 = 0.67, permutation p<0.001). Thus, early sham trial data could be used to estimate the variability of stimulation responses for power analysis calculations. We show that approximately 10 sham trials were needed to robustly estimate sham variability. Power analysis (using a paired-t test) showed that for effect sizes [&ge;] 1.1, roughly 10 trials should be used per stimulation site for sufficiently powered results.

ConclusionsThe presented workflow is adaptable to multiple indications and is specifically designed to overcome key challenges experienced during stimulation site testing. Through incorporating sham trials, effect size calculations, and tolerability testing, the described approach can be used to identify personalized and clinically efficacious stimulation sites.
]]></description>
<dc:creator>Saal, J.</dc:creator>
<dc:creator>Kadlec, K.</dc:creator>
<dc:creator>Allawala, A. B.</dc:creator>
<dc:creator>Johnston, L.</dc:creator>
<dc:creator>Leriche, R.</dc:creator>
<dc:creator>Vatsyayan, R.</dc:creator>
<dc:creator>Han, Y.</dc:creator>
<dc:creator>Kist, A.</dc:creator>
<dc:creator>Di Ianni, T.</dc:creator>
<dc:creator>Dawes, H. E.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Lee, A. M.</dc:creator>
<dc:creator>Krystal, A. D.</dc:creator>
<dc:creator>Moussawi, K.</dc:creator>
<dc:creator>Shirvalkar, P.</dc:creator>
<dc:creator>Sellers, K. K.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.22.649607</dc:identifier>
<dc:title><![CDATA[Towards individualized deep brain stimulation: A stereoencephalography-based workflow for unbiased neurostimulation target identification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.25.650293v1?rss=1">
<title>
<![CDATA[
Interrupting Elmsan1 repression of nuclear Acetyl-CoA production therapeutically reprograms cancer cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.25.650293v1?rss=1</link>
<description><![CDATA[
Metabolites are essential substrates for epigenetic modifications. Although nuclear acetyl-CoA constitutes a small fraction of the whole cell pool, it regulates cell fate by locally providing histone acetylation substrate. Here, we combined phenotypic chemical screen and genome-wide CRISPR screen to demonstrate a nucleus-specific acetyl-CoA regulatory mechanism that can be modulated to achieve therapeutic cancer cell reprogramming. While previously thought that nucleus-localized pyruvate dehydrogenase complex (nPDC) is constitutively active, we found that nPDC is constitutively inhibited by the nuclear protein ELMSAN1 through direct interaction. Pharmacologic inhibition of the ELMSAN1-nPDC interaction derepressed nPDC activity, enhancing nuclear acetyl-CoA generation and reprogramming cancer cells to a postmitotic state with diminished cell-of-origin signatures. Reprogramming was synergistically enhanced by histone deacetylase 1/2 inhibition, resulting in inhibited tumor growth, durably suppressed tumor-initiating ability, and improved survival in multiple cancer types in vivo, including therapy-resistant sarcoma patient-derived xenografts and carcinoma cell line xenografts. Our findings highlight the potential of targeting ELMSAN1-nPDC as epigenetic cancer therapy.
]]></description>
<dc:creator>Zhao, T.</dc:creator>
<dc:creator>He, L.</dc:creator>
<dc:creator>Wong, L. P.</dc:creator>
<dc:creator>Mei, S.</dc:creator>
<dc:creator>Xia, J.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Van Vranken, J.</dc:creator>
<dc:creator>Mazzola, M.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Rhee, C.</dc:creator>
<dc:creator>Fang, T.</dc:creator>
<dc:creator>Fukushima, T.</dc:creator>
<dc:creator>Sayles, L.</dc:creator>
<dc:creator>Diaz, M.</dc:creator>
<dc:creator>Gibbons, J. A. B.</dc:creator>
<dc:creator>Mostoslavsky, R.</dc:creator>
<dc:creator>Gygi, S. P.</dc:creator>
<dc:creator>Dou, Z.</dc:creator>
<dc:creator>Sykes, D. B.</dc:creator>
<dc:creator>Sadreyev, R.</dc:creator>
<dc:creator>Sweet-Cordero, A.</dc:creator>
<dc:creator>Scadden, D. T.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.25.650293</dc:identifier>
<dc:title><![CDATA[Interrupting Elmsan1 repression of nuclear Acetyl-CoA production therapeutically reprograms cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.26.650725v1?rss=1">
<title>
<![CDATA[
Discovery of a tRNA-regulatory transcription factor that suppresses breast cancer metastasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.26.650725v1?rss=1</link>
<description><![CDATA[
Transfer RNAs (tRNAs), once viewed as static adaptors in translation, are now recognized as dynamic regulators of gene expression. While recent studies have illuminated roles for tRNA stability, the upstream mechanisms governing tRNA transcription remain poorly understood. To address this gap, we generated the EXpression atlas of tRNA (EXTRNA), a high-resolution tRNA expression dataset spanning 24 cell lines across 9 human tissues. EXTRNA revealed both tissue-type-specific expression programs ("tRNAomes") and unexpected intra-tissue heterogeneity across breast cancer samples. Integrating EXTRNA with computational network analysis and data from other publicly available datasets, we identified Zinc Finger ZZ-Type And EF-Hand Domain Containing 1 (ZZEF1) as the first sequence-specific transcription factor of a particular tRNA. ZZEF1 promoted tRNA-LysUUU transcription by partnering with the ATP-dependent chromatin remodeler Chromodomain Helicase DNA Binding Protein 6 (CHD6), enhancing chromatin accessibility at tRNA-Lys-TTT-3 loci. ZZEF1 deficiency reduced tRNA-LysUUU abundance, decreased the translational efficiency of AAR codon-enriched mRNAs--including the tumor suppressor Serine/Threonine Kinase 3 (STK3)--and promoted metastatic progression in breast cancer in vivo. Together, our findings establish a previously unrecognized mechanism for RNA polymerase III-mediated tRNA transcription and define a regulatory circuit linking chromatin remodeling, codon-specific translation, and tumor suppression. More broadly, this work introduces a framework for dissecting the regulatory logic of the tRNAome and highlights tRNA expression control as a promising avenue for therapeutic intervention.
]]></description>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Markett, D.</dc:creator>
<dc:creator>Karimzadeh, M.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Khoroshkin, M. S.</dc:creator>
<dc:creator>Boyraz, B.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Carpenter, C.</dc:creator>
<dc:creator>Nguyen, P.</dc:creator>
<dc:creator>Garcia, K.</dc:creator>
<dc:creator>Joshi, T.</dc:creator>
<dc:creator>Martin, C.</dc:creator>
<dc:creator>Hanisch, B.</dc:creator>
<dc:creator>Molina, H.</dc:creator>
<dc:creator>Tavazoie, S.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Navickas, A.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.26.650725</dc:identifier>
<dc:title><![CDATA[Discovery of a tRNA-regulatory transcription factor that suppresses breast cancer metastasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.28.650977v1?rss=1">
<title>
<![CDATA[
ARAP2 regulates responses to interferon-gamma by restricting SOCS1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.28.650977v1?rss=1</link>
<description><![CDATA[
Interferon-gamma (IFN{gamma}) is critical for immunity against intra-macrophagic pathogens, signaling through the JAK-STAT pathway to induce a tyrosine-phosphorylation cascade that ensures a potent immune response. Excessive JAK-STAT signaling can drive hyperinflammation and autoimmunity, and thus signaling is tightly and selectively regulated by the IFN{gamma}-inducible protein, Suppressor of Cytokine Signaling 1 (SOCS1). SOCS1 inhibits signaling by directly blocking JAK kinase activity. Here we identified a SOCS1-interacting partner, ARAP2 that fine-tunes SOCS1 function. We report that tyrosine 415 in ARAP2 binds the SOCS1-Src Homology 2 (SH2) domain and limits the ability of SOCS1 to inhibit IFN{gamma} signaling. Our findings show that ARAP2 promotes the IFN{gamma} response through a phosphorylation dependent interaction with the negative regulator SOCS1.
]]></description>
<dc:creator>Keating, N.</dc:creator>
<dc:creator>Doggett, K.</dc:creator>
<dc:creator>Bidgood, G.</dc:creator>
<dc:creator>Guzman, L. G. M.</dc:creator>
<dc:creator>Dagley, L.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Gabrielyan, A.</dc:creator>
<dc:creator>Alvarado, C.</dc:creator>
<dc:creator>Williams, B.</dc:creator>
<dc:creator>Broomfield, B.</dc:creator>
<dc:creator>Duckworth, B.</dc:creator>
<dc:creator>Hockings, C.</dc:creator>
<dc:creator>Youssef, J.</dc:creator>
<dc:creator>Leong, E.</dc:creator>
<dc:creator>Morris, R.</dc:creator>
<dc:creator>Kueh, A.</dc:creator>
<dc:creator>Garnham, A.</dc:creator>
<dc:creator>Casanova, J.-L.</dc:creator>
<dc:creator>Boisson-Dupuis, S.</dc:creator>
<dc:creator>Babon, J.</dc:creator>
<dc:creator>Linossi, E.</dc:creator>
<dc:creator>Tate, M. D.</dc:creator>
<dc:creator>Groom, J. R.</dc:creator>
<dc:creator>Nicholson, S.</dc:creator>
<dc:date>2025-04-29</dc:date>
<dc:identifier>doi:10.1101/2025.04.28.650977</dc:identifier>
<dc:title><![CDATA[ARAP2 regulates responses to interferon-gamma by restricting SOCS1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.27.649460v1?rss=1">
<title>
<![CDATA[
Transcriptomic analysis reveals immune signatures associated with specific cutaneous manifestations of lupus in systemic lupus erythematosus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.27.649460v1?rss=1</link>
<description><![CDATA[
Systemic lupus erythematosus (SLE) presents with diverse heterogenous cutaneous manifestations. However, the molecular and immunologic pathways driving specific cutaneous manifestations of SLE are poorly understood. Here, we leverage transcriptomics from a large well-phenotyped longitudinal cohort of SLE patients to map molecular pathways linked to ten distinct SLE-related rashes. Through whole blood and immune cell-sorted bulk RNA sequencing, we identified immune signatures specific to cutaneous subtypes of SLE. Subacute cutaneous lupus (SCLE) exhibited broad upregulation of interferon, TNF-, and IL6-JAK-STAT3 pathways suggesting potential unique therapeutic responses to JAK and type I interferon inhibition. While interferon signaling is prominent in SCLE, discoid lupus, and acute lupus, it is unexpectedly attenuated in patients with skin and mucosal ulcers. Pathway and cell-type enrichment analysis revealed unexpected roles for CD14+ monocytes in photosensitivity of SLE and NK cells in alopecia, mucosal ulceration, and livedo reticularis. These findings illuminate the immune heterogeneity of rashes in SLE, highlighting subtype-specific mechanistic targets, and presenting opportunities for precision therapies for SLE-associated skin phenotypes.
]]></description>
<dc:creator>Lee, E. Y.</dc:creator>
<dc:creator>Patterson, S.</dc:creator>
<dc:creator>Cutts, Z.</dc:creator>
<dc:creator>Lanata, C. M.</dc:creator>
<dc:creator>Dall'Era, M.</dc:creator>
<dc:creator>Yazdany, J.</dc:creator>
<dc:creator>Criswell, L. A.</dc:creator>
<dc:creator>Haemel, A.</dc:creator>
<dc:creator>Katz, P.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2025-04-30</dc:date>
<dc:identifier>doi:10.1101/2025.04.27.649460</dc:identifier>
<dc:title><![CDATA[Transcriptomic analysis reveals immune signatures associated with specific cutaneous manifestations of lupus in systemic lupus erythematosus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.28.651083v1?rss=1">
<title>
<![CDATA[
Regulatory logic of human cortex evolution by combinatorial perturbations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.28.651083v1?rss=1</link>
<description><![CDATA[
Comparative genomic studies between contemporary and extinct hominins revealed key evolutionary modifications, but their number has hampered a system level investigation of their combined roles in scaffolding modern traits. Through multi-layered integration we selected 15 genes carrying nearly fixed sapiens-specific protein-coding mutations and developed a scalable design of combinatorial CRISPR-Cas9 bidirectional perturbations to uncover their regulatory hierarchy in cortical brain organoids. Interrogating the effects of overexpression and downregulation for all gene pairs in all possible combinations, we defined their impact on transcription and differentiation and reconstructed their regulatory architecture. We uncovered marked cell type-specific effects, including the promotion of alternative fates and the emergence of interneuron populations, alongside a core subnetwork comprising KIF15, NOVA1, RB1CC1 and SPAG5 acting as central regulator across cortical cell types.
]]></description>
<dc:creator>Skaros, A.</dc:creator>
<dc:creator>Vitriolo, A.</dc:creator>
<dc:creator>Leonardi, O.</dc:creator>
<dc:creator>Finazzi, V.</dc:creator>
<dc:creator>Pereira, M. F.</dc:creator>
<dc:creator>Prazzoli, F.</dc:creator>
<dc:creator>Trattaro, S.</dc:creator>
<dc:creator>Moriano, J.</dc:creator>
<dc:creator>Capocefalo, D.</dc:creator>
<dc:creator>Villa, C. E.</dc:creator>
<dc:creator>Boettcher, M.</dc:creator>
<dc:creator>Boeckx, C.</dc:creator>
<dc:creator>Testa, G.</dc:creator>
<dc:date>2025-05-01</dc:date>
<dc:identifier>doi:10.1101/2025.04.28.651083</dc:identifier>
<dc:title><![CDATA[Regulatory logic of human cortex evolution by combinatorial perturbations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.30.651576v1?rss=1">
<title>
<![CDATA[
Evolutionary Sequence and Structural Basis for the Epistatic Origins of Drug Resistance in HIV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.30.651576v1?rss=1</link>
<description><![CDATA[
The emergence of drug resistance in the human immunodeficiency virus (HIV) remains a formidable challenge to the long-term efficacy of antiretroviral therapy (ART). A growing body of evidence highlights the critical role of epistasis, the dependence of mutational effects on the sequence context, in shaping the fitness landscape of HIV under ART-induced selection pressure. However, the biophysical origins of the epistatic interactions involved in engendering drug-resistance mutations (DRMs) remain unclear. Are the mutational correlations "intrinsic" to the properties of the protein, or do they arise because of drug binding? We use a Potts sequence-covariation statistical energy model built on patient-derived HIV-1 protein sequences to construct computational double mutant cycles that probe pairwise epistasis for all observed mutations across the three major HIV drug-target enzymes. We find that the strongest epistatic effects occur between mutations at residue positions that frequently mutate during the course of ART, termed resistance-associated positions. To investigate the structural origins of the strongest epistatic interactions, we perform [~]100 free energy perturbation molecular dynamics simulations, revealing that the primary contribution to the pairwise epistasis between DRMs arises from cooperative effects on protein stability and folding as an intrinsic consequence of the protein mutational landscape. The results collectively reinforce a mechanism of resistance evolution whereby viruses escape drug pressure by selectively engendering mutations at "intrinsically" coupled sites, allowing them to cooperatively ameliorate fitness detriments incurred by individual DRMs.

SignificanceEpistasis refers to the phenomenon where the effect of a mutation on protein structure and function is dependent on the genetic sequence background of the mutation, resulting in the combined effect of mutations being non-additive. Epistasis plays a significant role in the evolution of drug resistance in viruses such as HIV under therapeutic selection pressure. We combine a protein sequence coevolutionary model and molecular dynamics free energy simulations to identify and probe the mechanistic origins of the strongest epistatic interactions connecting HIV drug-resistance mutations. The work establishes a foundation to probe the molecular bases of epistasis and predict the evolution of resistance predicated on the knowledge of epistatic interaction networks.
]]></description>
<dc:creator>Biswas, A.</dc:creator>
<dc:creator>Choudhuri, I.</dc:creator>
<dc:creator>Huang, K.</dc:creator>
<dc:creator>Sun, Q.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Haldane, A.</dc:creator>
<dc:creator>Levy, R.</dc:creator>
<dc:creator>Lyumkis, D.</dc:creator>
<dc:date>2025-05-02</dc:date>
<dc:identifier>doi:10.1101/2025.04.30.651576</dc:identifier>
<dc:title><![CDATA[Evolutionary Sequence and Structural Basis for the Epistatic Origins of Drug Resistance in HIV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.01.651538v1?rss=1">
<title>
<![CDATA[
An intrinsically disordered region of histone demethylase KDM5A activates catalysis through interactions with the nucleosomal acidic patch and DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.01.651538v1?rss=1</link>
<description><![CDATA[
Lysine demethylase 5A (KDM5A) plays a key role in the regulation of chromatin accessibility by catalyzing the removal of trimethyl marks on histone H3K4 (H3K4me3). KDM5A is also an oncogenic driver, with overexpression of KDM5A observed in various cancers, including breast, lung, and ovarian cancer. Past studies have characterized the functions of KDM5A domains, including KDM5A interactions with the histone H3 tail, but have yet to identify the broader mechanisms that drive KDM5A binding to the nucleosome. Through investigation of binding and catalysis on nucleosome substrates, we uncovered multivalent interactions of KDM5A with the H2A/H2B acidic patch and DNA that play crucial roles in the regulation of catalytic activity. We also identified an intrinsically disordered region (IDR) containing bifunctional arginine-rich motifs capable of binding to both the histone H2A/H2B acidic patch and nucleosomal DNA that is necessary for catalysis on nucleosome substrates. Our findings both elucidate previously unknown mechanisms that regulate KDM5A catalytic activity and reveal the ability of an IDR to engage in multiple interactions with chromatin.

ARTICLE HIGHLIGHTSO_LIThe intrinsically disordered region of KDM5A binds the acidic patch and DNA.
C_LIO_LIInteractions with the nucleosome are mediated by arginine-rich motifs in the IDR.
C_LIO_LIThe IDR properly orients KDM5A on the nucleosome to enable catalysis.
C_LI
]]></description>
<dc:creator>Palla, A. M.</dc:creator>
<dc:creator>Lin, C.-C.</dc:creator>
<dc:creator>Trnka, M. J.</dc:creator>
<dc:creator>Leao, E. M.</dc:creator>
<dc:creator>Petronikolou, N.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>McGinty, R. K.</dc:creator>
<dc:creator>Fujimori, D. G.</dc:creator>
<dc:date>2025-05-02</dc:date>
<dc:identifier>doi:10.1101/2025.05.01.651538</dc:identifier>
<dc:title><![CDATA[An intrinsically disordered region of histone demethylase KDM5A activates catalysis through interactions with the nucleosomal acidic patch and DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.29.651039v1?rss=1">
<title>
<![CDATA[
Harmonized Protocol for Subfield Segmentation in the Hippocampal Body on High-Resolution in vivo MRI from the Hippocampal Subfields Group (HSG) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.29.651039v1?rss=1</link>
<description><![CDATA[
Hippocampal subfields differentially develop and age, and they vary in vulnerability to neurodegenerative diseases. Innovation in high-resolution imaging has accelerated clinical research on human hippocampal subfields, but substantial differences in segmentation protocols impede comparisons of results across laboratories. The Hippocampal Subfields Group (HSG) is an international organization seeking to address this issue by developing a histologically-valid, reliable, and freely available segmentation protocol for high-resolution T2-weighted 3 tesla MRI (http://www.hippocampalsubfields.com). Here, we report the first portion of the protocol focused on subfields in the hippocampal body; protocols for the head and tail are in development. The body protocol includes definitions of the internal boundaries between subiculum, Cornu Ammonis (CA) 1-3 subfields, and dentate gyrus, in addition to the external boundaries of the hippocampus apart from surrounding white matter and cerebrospinal fluid. The segmentation protocol is based on a novel histological reference data set labeled by multiple expert neuroanatomists. With broad participation of the research community, we voted on the segmentation protocol via online survey, which included detailed protocol information, feasibility testing, demonstration videos, example segmentations, and labeled histology. All boundary definitions were rated as having high clarity and reached consensus agreement by Delphi procedure. The harmonized body protocol yielded high inter- and intra-rater reliability. In the present paper we report the procedures to develop and test the protocol, as well as the detailed procedures for manual segmentation using the harmonized protocol. The harmonized protocol will significantly facilitate cross-study comparisons and provide increased insight into the structure and function of hippocampal subfields across the lifespan and in neurodegenerative diseases.
]]></description>
<dc:creator>Daugherty, A. M.</dc:creator>
<dc:creator>Carr, V.</dc:creator>
<dc:creator>Canada, K.</dc:creator>
<dc:creator>Radman, G.</dc:creator>
<dc:creator>Brown, T.</dc:creator>
<dc:creator>Augustinack, J.</dc:creator>
<dc:creator>Amunts, K.</dc:creator>
<dc:creator>Bakker, A.</dc:creator>
<dc:creator>Berron, D.</dc:creator>
<dc:creator>Burggren, A.</dc:creator>
<dc:creator>Chetelat, G.</dc:creator>
<dc:creator>de Flores, R.</dc:creator>
<dc:creator>Ding, S.-L.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Insausti, R.</dc:creator>
<dc:creator>Johnson, E.</dc:creator>
<dc:creator>Kanel, P.</dc:creator>
<dc:creator>Keresztes, A.</dc:creator>
<dc:creator>Kedo, O.</dc:creator>
<dc:creator>Kennedy, K. M.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Malykhin, N.</dc:creator>
<dc:creator>Martinez, A.</dc:creator>
<dc:creator>Mueller, S.</dc:creator>
<dc:creator>Mulligan, E.</dc:creator>
<dc:creator>Ofen, N.</dc:creator>
<dc:creator>Palombo, D.</dc:creator>
<dc:creator>Pasquini, L.</dc:creator>
<dc:creator>Pluta, J.</dc:creator>
<dc:creator>Raz, N.</dc:creator>
<dc:creator>Riggins, T.</dc:creator>
<dc:creator>Rodrigue, K. M.</dc:creator>
<dc:creator>Saifullah, S.</dc:creator>
<dc:creator>Schlichting, M. L.</dc:creator>
<dc:creator>Stark, C.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Yushkevich, P.</dc:creator>
<dc:creator>La Joie, R.</dc:creator>
<dc:creator>Wisse, L.</dc:creator>
<dc:creator>Olsen, R.</dc:creator>
<dc:creator>the Alzheimers Disease Neuroimaging Initiative,</dc:creator>
<dc:creator>the Hippocampal Subfields Gro</dc:creator>
<dc:date>2025-05-04</dc:date>
<dc:identifier>doi:10.1101/2025.04.29.651039</dc:identifier>
<dc:title><![CDATA[Harmonized Protocol for Subfield Segmentation in the Hippocampal Body on High-Resolution in vivo MRI from the Hippocampal Subfields Group (HSG)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.01.651659v1?rss=1">
<title>
<![CDATA[
Antibody response to Aedes aegypti D7L1+2 salivary proteins as marker of aggregate vector exposure and correlate of dengue virus susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.01.651659v1?rss=1</link>
<description><![CDATA[
Aedes aegypti mosquitoes transmit several arboviruses of public health importance. Among these is dengue virus (DENV), a flavivirus whose global infection rates continue to rise each year. With limited options available for preventing or treating DENV infections, mosquito control remains the most widely implemented strategy to combat DENV transmission. Due to the global distribution of DENV, which infects an estimated 400 million people per year, vector suppression practices vary drastically by country and/or region and even small differences in microenvironment can significantly impact vector abundance. There remains a significant need to better understand vector exposure rates at an individual level to disentangle vector exposure and arboviral infection rates. To this end, we have optimized a serologic assay to assess the abundance of antibodies directed against the mosquito salivary proteins AeD7L1+2 as a surrogate metric of vector exposure. Utilizing this assay, we found that anti-D7L1+2 IgG levels were unable to identify low levels of Aedes exposure in individuals with limited prior Aedes exposure, indicating they are unreliable markers of an individuals recent exposure to low levels of these vectors. However, antibody levels against D7L1+2 were robust in plasma samples from individuals living in Aedes endemic regions. These antibody levels reflected seasonal changes in Aedes abundance and exposure, indicating their potential for use as an aggregate marker of vector exposure. Additionally, we found that there were slight negative associations with anti-D7L1+2 IgG levels and age in our cohort. Interestingly, we also found that lower titers of anti-AeD7L1+2 IgG correlated with higher infection burden in households. This finding has implications for the potential interaction between AeD7 proteins and DENV during infection events that will require further study.
]]></description>
<dc:creator>Bahr, L.</dc:creator>
<dc:creator>Hamins-Puertolas, M.</dc:creator>
<dc:creator>Buddhari, D.</dc:creator>
<dc:creator>Petzlova, I.</dc:creator>
<dc:creator>Oliveira, F.</dc:creator>
<dc:creator>Thomas, S. J.</dc:creator>
<dc:creator>Thangamani, S.</dc:creator>
<dc:creator>Waickman, A.</dc:creator>
<dc:creator>Anderson, K.</dc:creator>
<dc:date>2025-05-06</dc:date>
<dc:identifier>doi:10.1101/2025.05.01.651659</dc:identifier>
<dc:title><![CDATA[Antibody response to Aedes aegypti D7L1+2 salivary proteins as marker of aggregate vector exposure and correlate of dengue virus susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.04.652125v1?rss=1">
<title>
<![CDATA[
A Cluster of Acidic Residues in the Cytoplasmic Domain of SARS-CoV-2 Spike is Required for Virion-Incorporation and Infectivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.04.652125v1?rss=1</link>
<description><![CDATA[
Abstract/SummaryLike all coronaviruses, the infectivity of SARS-CoV-2 virus particles (virions) requires incorporation of the Spike glycoprotein. Yet, the mechanisms that support the virion-incorporation of Spike are incompletely defined. We noted an unusual feature of human sarbecovirus Spike proteins: their cytoplasmic domains (CDs) contain a stretch of acidic amino acids (DEDDSE). This sequence resembles a cluster of acidic residues, or acidic cluster (AC) motif, found in the cytoplasmic domain of the cellular endoprotease Furin. In Furin, the acidic cluster acts as a protein sorting signal, supporting its intracellular localization at the trans-Golgi network (TGN). We tested the contribution of the acidic cluster motif in the Spike CD to protein interactions and to the infectivity of SARS-CoV-2. We used virus-like particles (VLPs) as a model for viral "infection" (transduction). The SARS-CoV2 VLPs were produced by co-expressing Spike (S), Membrane (M), Envelope (E) and Nucleocapsid (N) proteins and deliver an RNA encoding luciferase to target cells expressing the ACE2 receptor. Remarkably, when all five acidic residues of the DEDDSE sequence were replaced with alanines, the VLPs were rendered non-infectious. The N-terminal DE residues provided most of the activity of the acidic cluster. These virologi-cally-impaired Spike mutants were able to reach the cell surface and induce the formation of syncytia, indicating that they are fusogenic and capable of anterograde traffic through the biosynthetic pathway to the plasma membrane. Despite this, they failed to efficiently incorporate into virions. We observed acidic cluster motif-dependent interactions of the Spike CD with several cellular proteins that could potentially support its role in virion-incorporation, including the ERM proteins Ezrin, Radixin, and Moesin; the retromer subunit Vps35, and the medium subunits of the clathrin adaptor complexes AP1 and AP2. While the key cofactor and mechanism of action remains to be defined, this region of acidic residues in the Spike CD appears to be a novel determinant of SARS-CoV-2 infectivity.
]]></description>
<dc:creator>Stoneham, C. A.</dc:creator>
<dc:creator>Singh, R.</dc:creator>
<dc:creator>De Leon, A.</dc:creator>
<dc:creator>Tafelmeyer, P.</dc:creator>
<dc:creator>Acosta, F.</dc:creator>
<dc:creator>Fuori, A.</dc:creator>
<dc:creator>Anderson, M.</dc:creator>
<dc:creator>Ramirez, P. W.</dc:creator>
<dc:creator>Schwartzer-Sperber, H. S.</dc:creator>
<dc:creator>Pillai, S. K.</dc:creator>
<dc:creator>Lewinski, M. K.</dc:creator>
<dc:creator>Guatelli, J.</dc:creator>
<dc:date>2025-05-06</dc:date>
<dc:identifier>doi:10.1101/2025.05.04.652125</dc:identifier>
<dc:title><![CDATA[A Cluster of Acidic Residues in the Cytoplasmic Domain of SARS-CoV-2 Spike is Required for Virion-Incorporation and Infectivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.05.651928v1?rss=1">
<title>
<![CDATA[
Investigating Local Negative Feedback of Rac Activity by Mathematical Models and Cell Motility Simulations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.05.651928v1?rss=1</link>
<description><![CDATA[
For polarization and directed migration, cells use a combination of local positive feedback and long-range inhibition. We have previously used mathematical models to show the ability of this core circuit to regulate directed cell movement. However, this wave pinning model lacks important additional feedback circuits, including the recently demonstrated local negative feedback from Town and Weiner. Here we extend our models to investigate the consequences of this additional link on cell physiology. We model responses of neutrophil-like HL-60 cells to spatially-controlled optogenetic stimulation of PI3K, leading (via PIP3) to Rac activity. We sequentially build up and investigate partial differential equation (PDE) models of the key Rac, Rac-Inhibitor, and PIP3-Rac-Inhibitor circuits. We fit model parameters to temporal and spatial (cell trajectory) data. Cell shapes, motility, and responses to stimuli are modeled in 2D cell-based simulations, with PDEs for Rac and the other regulatory components solved along the cell edge. We demonstrate that the ability of modeled cells to respond to temporal as well as spatial features of guidance cues depends on the addition of the local negative feedback circuit. Furthermore, the local Rac inhibitor improves the ability of modeled cells to respond to noisy or dynamic extracellular gradients. Our work demonstrates how local negative feedback enhances dynamic polarity and gradient sensing in migratory cells.
]]></description>
<dc:creator>Algorta, J.</dc:creator>
<dc:creator>Town, J. P.</dc:creator>
<dc:creator>Weiner, O.</dc:creator>
<dc:creator>Edelstein-Keshet, L.</dc:creator>
<dc:date>2025-05-06</dc:date>
<dc:identifier>doi:10.1101/2025.05.05.651928</dc:identifier>
<dc:title><![CDATA[Investigating Local Negative Feedback of Rac Activity by Mathematical Models and Cell Motility Simulations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.30.651490v1?rss=1">
<title>
<![CDATA[
Identification of a RIPK2-Regulated Gene Signature as a Candidate Biomarker for RIPK2 Activity and Prognosis in Prostate Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.30.651490v1?rss=1</link>
<description><![CDATA[
Receptor-interacting protein kinase 2 (RIPK2) has emerged as a promising drug target in various cancers, including prostate cancer (PC). However, the absence of reliable biomarkers to assess RIPK2 activity limits both patient selection for anti-RIPK2 therapies and treatment monitoring. To address this gap, we performed RNA-Seq analysis on PC cell lines (22Rv1, DU145, and PC3) with CRISPR/Cas9-mediated RIPK2 knockout (RIPK2-KO) using two independent guide RNAs. This analysis identified 13 candidate RIPK2-regulated genes, of which eight were validated by reverse transcription quantitative PCR (RT-qPCR). Furthermore, treatment with two distinct RIPK2 inhibitors significantly reduced RIPK2 signature scores in five independent PC cell lines in a dose- and/or time-dependent manner. Clinical association analyses revealed that high RIPK2 signature scores correlate with metastasis and worse biochemical recurrence-free, progression-free, disease-free, and overall survival, outperforming RIPK2 mRNA levels as a prognostic biomarker. This study establishes, for the first time, a RIPK2-regulated gene signature as a potential biomarker for RIPK2 activity and PC prognosis, warranting further validation in clinical specimens to provide a much-needed tool for patient stratification and response monitoring in RIPK2-targeted therapies.
]]></description>
<dc:creator>Elgehama, A. M.</dc:creator>
<dc:creator>Yang, Q.</dc:creator>
<dc:creator>He, Z.</dc:creator>
<dc:creator>Ruegg, L. E.</dc:creator>
<dc:creator>You, S.</dc:creator>
<dc:creator>Yang, W.</dc:creator>
<dc:date>2025-05-07</dc:date>
<dc:identifier>doi:10.1101/2025.04.30.651490</dc:identifier>
<dc:title><![CDATA[Identification of a RIPK2-Regulated Gene Signature as a Candidate Biomarker for RIPK2 Activity and Prognosis in Prostate Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.01.651686v1?rss=1">
<title>
<![CDATA[
Detecting neuroplastic effects induced by ketamine in healthy human subjects: a multimodal approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.01.651686v1?rss=1</link>
<description><![CDATA[
We investigated ketamines neuroplastic effects in healthy human subjects using integrated Positron Emission Tomography (PET)/Magnetic Resonance Imaging (MRI) measures before and 1-8 days after a single psychedelic dose of ketamine (1 mg/kg, intravenous). Eleven participants underwent two PET/MRI scans with [11C]-UCBJ (synaptic density/plasticity), 1H-MRS (Glutamate and GABA), and resting-state fMRI (intrinsic brain activity, functional connectivity, graph-theoretic metrics), before and after ketamine. While group-level analyses showed only trend-level increases in PET synaptic markers, we observed significantly elevated Anterior Cingulate Cortex (ACC) glutamate levels post-ketamine. Functional connectivity analyses revealed decreased within-network integrity, particularly in high-order networks like the default mode network (DMN), alongside increased low-to-high-order network integration. Our multimodal analysis showed that increased [11C]-UCBJ volume distribution (VT), a putative index of synaptic plasticity, correlated with reduced intrinsic activity in DMN regions and decreased influence of the posterior cingulate cortex (PCC) in global network dynamics. By linking molecular and network-level changes, our results point to the PCC as a central hub where ketamine may reshape brain hierarchies in the long term, providing new directions for understanding its therapeutic mechanisms and developing targeted treatments.
]]></description>
<dc:creator>Agnorelli, C.</dc:creator>
<dc:creator>Peill, J.</dc:creator>
<dc:creator>Sawicka, G.</dc:creator>
<dc:creator>Kurtin, D. L.</dc:creator>
<dc:creator>Shatalina, E.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Wall, M. B.</dc:creator>
<dc:creator>Rua, C.</dc:creator>
<dc:creator>Godfrey, K.</dc:creator>
<dc:creator>Ertl, N.</dc:creator>
<dc:creator>Searle, G.</dc:creator>
<dc:creator>Zhou, K.</dc:creator>
<dc:creator>Weiss, B.</dc:creator>
<dc:creator>Fagiolini, A.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Matthews, P. M.</dc:creator>
<dc:creator>Rabiner, E.</dc:creator>
<dc:creator>Nutt, D.</dc:creator>
<dc:creator>Erritzoe, D.</dc:creator>
<dc:date>2025-05-07</dc:date>
<dc:identifier>doi:10.1101/2025.05.01.651686</dc:identifier>
<dc:title><![CDATA[Detecting neuroplastic effects induced by ketamine in healthy human subjects: a multimodal approach]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.06.651945v1?rss=1">
<title>
<![CDATA[
Synaptic vesicles that store monoamines and glutamate differ in protein composition. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.06.651945v1?rss=1</link>
<description><![CDATA[
Neuromodulators such as the monoamines are known to differ from classical neurotransmitters like glutamate in the time scale of signaling due to activation of slower G protein-coupled receptors. Recent work has suggested that the mode of release also differs between classical and modulatory transmitters. Although many components of neurotransmitter release machinery have been identified, we still understand little about the mechanisms responsible for differences in release. In this study, we address the differences between release of dopamine and glutamate by comparing the composition of synaptic vesicles (SVs) that contain the vesicular monoamine transporter 2 (VMAT2) versus vesicular glutamate transporter 2 (VGLUT2). Previous work has shown that these SV populations differ in frequency dependence, recycling kinetics and biogenesis. Taking advantage of a CRISPR-generated knock-in mouse with a cytoplasmic hemagglutinin (HA) tag at the N-terminus of VMAT2 to immunoisolate monoamine SVs, we find differences in the abundance and isoform expression of many SV protein families. Validation in primary neurons and in brain tissue confirms these differences in SV protein abundance between dopamine and glutamate release sites. Functional analysis reveals that the loss of differentially expressed SCAMP5 selectively impairs the recycling of VGLUT2 SVs, sparing VMAT2 vesicles in the same neuronal population. These findings provide new insights into the molecular diversity of SVs and the mechanisms that regulate the release of dopamine and glutamate, with implications for the physiological role of these transmitters and behavior.
]]></description>
<dc:creator>Asmerian, H.</dc:creator>
<dc:creator>Diaz, A. J.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>Alberts, J.</dc:creator>
<dc:creator>Sanetra, A. M.</dc:creator>
<dc:creator>Gnanabharathi, B.</dc:creator>
<dc:creator>Carr, N.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Edwards, R. H.</dc:creator>
<dc:creator>Silm, K.</dc:creator>
<dc:date>2025-05-07</dc:date>
<dc:identifier>doi:10.1101/2025.05.06.651945</dc:identifier>
<dc:title><![CDATA[Synaptic vesicles that store monoamines and glutamate differ in protein composition.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.07.652756v1?rss=1">
<title>
<![CDATA[
Associations between floor material and E. coli contamination in rural Bangladeshi households 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.07.652756v1?rss=1</link>
<description><![CDATA[
Soil floors are common in low-income countries and can harbor contamination from unsafely managed human and animal fecal waste. Soil/dust ingestion directly from floors or indirectly via hands, drinking water and food can significantly contribute to childrens ingestion of fecal organisms. We assessed if finished (e.g., concrete) floors are associated with lower E. coli contamination in the domestic environment in rural Bangladesh. We collected samples from 1864 households over 3.5 years, including stored drinking water, child and caregiver hand rinses, courtyard soil, food, and flies (n=24,118 samples), and enumerated E. coli using IDEXX Quanti-Tray/2000. Controlling for potential confounders (socio-demographics, water/sanitation status, animal ownership), households with finished floors had slightly lower log10-transformed E. coli counts ({Delta}log10= -0.10 (-0.20, 0.00)) and prevalence (prevalence ratio=0.90 (0.83, 0.98)) on child hands than households with soil floors; floor material was not associated with contamination levels in other sample types. Finished floors were associated with lower E. coli contamination of child hands, food and stored drinking water following periods of higher rainfall and temperature, and lower E. coli contamination of child hands in households with more domestic animals. Measures to control enteric infections in low-income countries should test flooring improvements to reduce exposure to fecal contamination.
]]></description>
<dc:creator>Tazin, S.</dc:creator>
<dc:creator>Islam, M.</dc:creator>
<dc:creator>Pickering, A. J.</dc:creator>
<dc:creator>Kwong, L. H.</dc:creator>
<dc:creator>Mertens, A.</dc:creator>
<dc:creator>Niven, C.</dc:creator>
<dc:creator>Arnold, B. F.</dc:creator>
<dc:creator>Hubbard, A. E.</dc:creator>
<dc:creator>Alam, M.</dc:creator>
<dc:creator>Sen, D.</dc:creator>
<dc:creator>Islam, S.</dc:creator>
<dc:creator>Rahman, M.</dc:creator>
<dc:creator>Unicomb, L.</dc:creator>
<dc:creator>Luby, S. P.</dc:creator>
<dc:creator>Benjamin-Chung, J.</dc:creator>
<dc:creator>Ercumen, A.</dc:creator>
<dc:date>2025-05-08</dc:date>
<dc:identifier>doi:10.1101/2025.05.07.652756</dc:identifier>
<dc:title><![CDATA[Associations between floor material and E. coli contamination in rural Bangladeshi households]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.04.652129v1?rss=1">
<title>
<![CDATA[
BRAFV600E-Driven Lung Tumorigenesis Requires Ligand-Mediated Activation of ERBB Receptor Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.04.652129v1?rss=1</link>
<description><![CDATA[
Secretion of ligands of the human epidermal growth factor (EGFR) family of receptors or erythroblastic leukemia viral oncogene family (ERBB1-4) is a feature common to many cancer cells. However, our understanding of the role of autocrine ligands in the aberrant behavior of cancer remains incomplete. Here we demonstrate that, in numerous preclinical models of lung tumorigenesis, BRAFV600E signaling promotes expression of ligands including HB-EGF, TGF, Epi- and Amphiregulin. Moreover, using both genetic or pharmacological approaches, we demonstrate that ligand-mediated activation of EGFR signaling in the tumor cell is required to sustain both early-stage BRAFV600E-driven lung tumorigenesis and supports late-stage BRAFV600E-driven lung cancer maintenance. Unbiased Reverse Phase Protein Analyses (RPPA) analyses, paired with targeted validation, reveals ERBB signaling serves to sustain signaling through the ERK1/2 MAP kinase pathway, through effects on ARAF and CRAF, and on the parallel JUN kinase (JNK) pathway. Furthermore, EGFR is activated in a cohort of BRAF-mutated lung cancer patients both pre- and post-treatment. Finally, we noted significant improvement in the depth and durability of therapeutic responses in preclinical models of BRAFV600E-driven lung cancer by combined inhibition of both BRAFV600E signaling plus pan-ERBB signaling. Collectively, this work provides evidence for an important role for ERBB family signaling in the genesis and maintenance of BRAFV600E-driven lung cancers, and the potential for future therapeutic improvement by rational combination targeting of these pathways.

SIGNIFICANCEBRAFT1799A serves as a predictive biomarker for FDA-approved targeted inhibition of BRAFV600E oncoprotein kinase signaling in non-small cell lung cancer (NSCLC). However the occurrence of primary or acquired drug resistance limit the depth and durability of patient responses. Studies described here provide a mechanistic rationale for clinical testing of first-line BRAFV600E inhibition combined with pan-ERBB inhibition to improve the depth and durability of initial patient responses, and delay the emergence of lethal drug resistant disease.
]]></description>
<dc:creator>Dacheux, M.</dc:creator>
<dc:creator>Wu, M.-J.</dc:creator>
<dc:creator>Scherzer, M. T.</dc:creator>
<dc:creator>Nillson, M.</dc:creator>
<dc:creator>Murphy, B.</dc:creator>
<dc:creator>Schuman, S.</dc:creator>
<dc:creator>Ju, Z.</dc:creator>
<dc:creator>Bivona, T.</dc:creator>
<dc:creator>Lito, P.</dc:creator>
<dc:creator>Gumbleton, M.</dc:creator>
<dc:creator>Puri, S.</dc:creator>
<dc:creator>Akerley, W.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Heymach, J. V.</dc:creator>
<dc:creator>Kinsey, C. G.</dc:creator>
<dc:creator>Negrao, M. V.</dc:creator>
<dc:creator>McMahon, M.</dc:creator>
<dc:creator>Vaishnavi, A.</dc:creator>
<dc:date>2025-05-09</dc:date>
<dc:identifier>doi:10.1101/2025.05.04.652129</dc:identifier>
<dc:title><![CDATA[BRAFV600E-Driven Lung Tumorigenesis Requires Ligand-Mediated Activation of ERBB Receptor Signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.05.652209v1?rss=1">
<title>
<![CDATA[
Genome Dynamics and Chromosome Structural Variations in Histoplasma ohiense, a fungal pathogen of humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.05.652209v1?rss=1</link>
<description><![CDATA[
Histoplasma is a clinically important but understudied genus of thermally dimorphic human fungal pathogens. Histoplasma species normally transition between a multicellular sporulating hyphal form in the soil and a unicellular pathogenic yeast form in a mammalian host. Little is known about genome plasticity of Histoplasma, which we address in this study with the ultimate goal of increasing our understanding of its pathogenicity. This study addresses the Histoplasma genome and its plasticity to further our understanding Histoplasmas ability to cause severe disease. Here we present the first telomere-to-telomere genome assemblies for Histoplasma ohiense. To analyze genome alterations between isolates, we develop an analysis tool to identify genome discontinuities relative to a reference genome. We utilize this analysis to interrogate the genome of laboratory strains and natural Histoplasma isolates and discover that the previously published reference genome does not completely match the chromosome structure of the majority of isolates, instead harboring reciprocal chromosome translocations. We identify the telomere-to-telomere Histoplasma ohiense reference genome that is most representative of clinical isolates. Additionally, to determine the rate of Histoplasma genomic changes, we sequence 46 passaged isolates and calculate the mutation rate to be 2.6 x10-10 SNP/base/doubling--the first such measurement to our knowledge within the order Onygenales, which encompasses several critical fungal pathogens. Finally, we sequence populations of cells to assess genome stability over the course of a month under both yeast and hyphal growth conditions. Interestingly, we observe that transposon signal is not static over time and instead increases during growth in both the yeast and hyphal forms as well as through morphologic transitions, suggesting an increase in transposon copy number. Taken together, this work highlights the plasticity of the Histoplasma genome and presents a comprehensive genome assembly that is representative of Histoplasma ohiense natural isolates.
]]></description>
<dc:creator>Heater, S.</dc:creator>
<dc:creator>Voorhies, M.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:date>2025-05-09</dc:date>
<dc:identifier>doi:10.1101/2025.05.05.652209</dc:identifier>
<dc:title><![CDATA[Genome Dynamics and Chromosome Structural Variations in Histoplasma ohiense, a fungal pathogen of humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.05.652301v1?rss=1">
<title>
<![CDATA[
The phosphatases TCPTP, PTPN22, and SHP1 play unique roles in T cell phosphotyrosine maintenance and feedback regulation of the TCR 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.05.652301v1?rss=1</link>
<description><![CDATA[
The protein tyrosine phosphatases (PTPs) TCPTP, PTPN22, and SHP1 are critical regulators of the activating phosphotyrosine (pY) site on the initiating T cell kinase, LckY394. Still, the broader implications of these phosphatases in T cell receptor (TCR) signalling and T cell biology remain unclear. By combining CRISPR/Cas9 gene editing and mass spectrometry, we evaluate the protein- and pY-level effects of TCPTP, PTPN22, and SHP1 in the Jurkat T cell model system. We find that deletion of each phosphatase corresponds to unique changes in the proteome of T cells, with few large-scale changes to TCR signalling proteins. Notably, PTPN22 and SHP1 deletions have opposing effects on pY abundance globally, while TCPTP deletion modestly elevates pY levels. Finally, we show that TCPTP is indirectly involved in Erk1/2 positive feedback to the TCR. Overall, our work provides evidence for alternative functions of three T cell phosphatases long thought to be redundant.
]]></description>
<dc:creator>Callahan, A.</dc:creator>
<dc:creator>Mojumdar, A.</dc:creator>
<dc:creator>Hu, M.</dc:creator>
<dc:creator>Wang, A.</dc:creator>
<dc:creator>Griffith, A.</dc:creator>
<dc:creator>Huang, N.</dc:creator>
<dc:creator>Chua, X. Y.</dc:creator>
<dc:creator>Mroz, N. M.</dc:creator>
<dc:creator>Puterbaugh, R. Z.</dc:creator>
<dc:creator>Reilly, S. P.</dc:creator>
<dc:creator>Salomon, A. R.</dc:creator>
<dc:date>2025-05-09</dc:date>
<dc:identifier>doi:10.1101/2025.05.05.652301</dc:identifier>
<dc:title><![CDATA[The phosphatases TCPTP, PTPN22, and SHP1 play unique roles in T cell phosphotyrosine maintenance and feedback regulation of the TCR]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.05.652217v1?rss=1">
<title>
<![CDATA[
Separable global and local beta burst dynamics in motor cortex of primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.05.652217v1?rss=1</link>
<description><![CDATA[
Sensorimotor beta band oscillations are known to modulate during normal movement control and abnormal beta modulation is linked to pathological bradykinesia. However, the functional differences between beta localized to one brain area versus beta synchronized across brain areas remains unclear. We monitored beta bursts in non-human primates, both neurotypical and stroke- impaired, during the performance of complex motor tasks. Across both groups of animals, we identified two distinct beta burst types: global bursts that tend to be synchronized across cortical and subcortical areas, and local bursts that tend to be confined to cortex. These two types exhibited distinct neural dynamics, with global bursts linked to reduced firing variability and overall slowed movements. In contrast, local bursts often occurred during the execution of complex behaviors, particularly during prehension. We found evidence for changes in the distribution of global and local bursts during recovery after stroke. In impaired animals early after stroke, global bursts predominated and were associated with reduced speed and impaired grasping. Notably, recovery of grasping was associated with a reduction in global bursts and an increase in local bursts, suggesting that local bursts may play an important role during prehension. Our findings reveal distinct roles of global and local beta bursts and indicate that the normalization of global and local burst timing tracks recovery of dexterity.
]]></description>
<dc:creator>Khanna, P.</dc:creator>
<dc:creator>farrokhi, b.</dc:creator>
<dc:creator>Choi, H.</dc:creator>
<dc:creator>Griffin, S.</dc:creator>
<dc:creator>Heimbuch, I.</dc:creator>
<dc:creator>Novik, L.</dc:creator>
<dc:creator>Thiesen, K.</dc:creator>
<dc:creator>Morrison, J. H.</dc:creator>
<dc:creator>Morecraft, R.</dc:creator>
<dc:creator>Ganguly, K.</dc:creator>
<dc:date>2025-05-10</dc:date>
<dc:identifier>doi:10.1101/2025.05.05.652217</dc:identifier>
<dc:title><![CDATA[Separable global and local beta burst dynamics in motor cortex of primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.06.652526v1?rss=1">
<title>
<![CDATA[
Transcription Factor (TF) validation using Dam-IT simultaneously captures genome-wide TF-DNA binding, direct gene regulation, and chromatin accessibility in plant cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.06.652526v1?rss=1</link>
<description><![CDATA[
Transcription Factors (TFs) govern vast networks of gene regulation. However, TF-DNA binding and TF-gene regulation datasets are typically measured separately due to experimental constraints, making it challenging to disentangle true biological relationships from batch effects. To fill this gap, we developed DamID-seq Incorporating Transcriptomics (Dam-IT), which simultaneously captures TF-DNA binding, direct TF-gene regulation, and chromatin accessibility in the same batch of cells. Dam-IT uses a transient cell-based TF-target validation system that is scalable and flexible to many experimental designs. As proof of concept, we used Dam-IT to reveal that bZIP1 directly regulates genes by binding to DNA regions of relatively low chromatin accessibility, supporting a "Hit-and-Run" mechanism of transcription.
]]></description>
<dc:creator>Hinckley, W. E.</dc:creator>
<dc:creator>Jack, A.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Frangos, S.</dc:creator>
<dc:creator>Pasquino, A.</dc:creator>
<dc:creator>Huang, S.-s. C.</dc:creator>
<dc:creator>Coruzzi, G. M.</dc:creator>
<dc:date>2025-05-11</dc:date>
<dc:identifier>doi:10.1101/2025.05.06.652526</dc:identifier>
<dc:title><![CDATA[Transcription Factor (TF) validation using Dam-IT simultaneously captures genome-wide TF-DNA binding, direct gene regulation, and chromatin accessibility in plant cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.09.653165v1?rss=1">
<title>
<![CDATA[
UNCOVERseq Enables Sensitive and Controlled Gene Editing Off-Target Nomination Across CRISPR-Cas Modalities and Systems 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.09.653165v1?rss=1</link>
<description><![CDATA[
The rapid development of CRISPR-Cas gene editing technologies has revolutionized genetic medicine, offering unprecedented precision and potential for treating a wide array of genetic disorders. However, assessing the risks of unintended gene editing effects remains critical, and is complicated by new editing modalities and unclear analytical guidelines. We present UNCOVERseq (Unbiased Nomination of CRISPR Off-target Variants using Enhanced RhPCR), an improved in cellulo off-target nomination workflow designed to sensitively nominate off-target sites (<0.01% editing) with defined input requirements and analytical process controls to provide empirical performance evidence across diverse circumstances. Using this workflow, we nominated off-targets across 192 guide RNAs (gRNAs) and demonstrated superior performance compared to existing methodologies. We identified a subset of six gRNAs with a dynamic range of specificity and confirmed the relevance and high true positive rate of our nomination method, providing relative risk assessments for multiple modalities (S.p. Cas9 and derived high-fidelity variants / base editors) in a translational system involving hematopoietic stem and progenitor cells (HSPCs). Additionally, we established that double-strand break (DSB) editing retains a strong, positive rank correlation to single-strand break (SSB)-mediated base editing, highlighting the importance of DSB nomination sites as candidate loci for base editing. Overall, UNCOVERseq improves informed risk assessment of gene editing in translational systems by enhancing the quality of off-target nomination.
]]></description>
<dc:creator>Kinney, K. J.</dc:creator>
<dc:creator>Jia, K.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Schmaljohn, E.</dc:creator>
<dc:creator>Osborne, T.</dc:creator>
<dc:creator>Thommandru, B.</dc:creator>
<dc:creator>Murugan, K.</dc:creator>
<dc:creator>Sanchez-Pena, A.</dc:creator>
<dc:creator>West, S.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Codipilly, R.</dc:creator>
<dc:creator>Sturgeon, M.</dc:creator>
<dc:creator>Turk, R.</dc:creator>
<dc:creator>McNeill, M.</dc:creator>
<dc:creator>Behlke, M.</dc:creator>
<dc:creator>Jacobi, A. M.</dc:creator>
<dc:creator>Cromer, K.</dc:creator>
<dc:creator>Kurgan, G.</dc:creator>
<dc:date>2025-05-11</dc:date>
<dc:identifier>doi:10.1101/2025.05.09.653165</dc:identifier>
<dc:title><![CDATA[UNCOVERseq Enables Sensitive and Controlled Gene Editing Off-Target Nomination Across CRISPR-Cas Modalities and Systems]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.12.652147v1?rss=1">
<title>
<![CDATA[
In utero lipid nanoparticle delivery achieves robust editing in hematopoietic stem cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.12.652147v1?rss=1</link>
<description><![CDATA[
In vivo genome editing for hematologic malignancies is limited by inefficient delivery of genome editors to hematopoietic stem cells (HSC) in the bone marrow. To overcome this limitation, we capitalized on the inherent liver tropism of lipid nanoparticles (LNPs) and the liver niche of fetal HSCs. We demonstrate that in utero delivery of LNPs without active targeting ligands to the fetal liver results in potentially therapeutic levels of HSC editing.
]]></description>
<dc:creator>Worthington, A. K.</dc:creator>
<dc:creator>Borges, B.</dc:creator>
<dc:creator>Lum, T.</dc:creator>
<dc:creator>Echeverri, E. S.</dc:creator>
<dc:creator>Zada, F. M.</dc:creator>
<dc:creator>Cordero, M.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Zenhausern, R.</dc:creator>
<dc:creator>Celik, O.</dc:creator>
<dc:creator>Shaw, C.</dc:creator>
<dc:creator>Gutierrez-Martinez, P.</dc:creator>
<dc:creator>Omarova, M.</dc:creator>
<dc:creator>Blanchard, C.</dc:creator>
<dc:creator>Burns, S.</dc:creator>
<dc:creator>Cromer, K.</dc:creator>
<dc:creator>Dahlman, J.</dc:creator>
<dc:creator>MacKenzie, T.</dc:creator>
<dc:date>2025-05-12</dc:date>
<dc:identifier>doi:10.1101/2025.05.12.652147</dc:identifier>
<dc:title><![CDATA[In utero lipid nanoparticle delivery achieves robust editing in hematopoietic stem cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.07.652676v1?rss=1">
<title>
<![CDATA[
A protective role of REM sleep in the pathological basal ganglia-cortical circuit of Parkinson's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.07.652676v1?rss=1</link>
<description><![CDATA[
Sleep disturbances have been shown to be intimately and bidirectionally related to disease progression across a wide range of neurodegenerative disorders including Parkinsons disease (PD) and Alzheimers disease. However, the precise neurophysiological mechanisms relating abnormal sleep to aberrant daytime network activity that accelerates disease progression has yet to be determined. We collected chronic, multi-night (n=40), intracranial cortico-basal recordings during sleep from a cohort of patients with PD along with paired polysomnography and morning self-reports. This revealed that longer duration (and shorter latency) of rapid eye movement (REM) sleep predicted reduced daytime resting beta (13-30 Hz) activity and cortico-basal functional and effective connectivity, features established to be pathophysiological in PD. Within REM sleep, stronger cortical delta activity specifically predicted reduced pathophysiological cortico-basal neural network features. Additionally, REM delta power significantly predicted greater self-reported morning alertness. These findings highlight a potentially protective role of REM sleep in cortico-basal network health in PD and daytime subjective experience, representing a potential target for closed loop neuromodulation therapies to impact neurodegenerative disease progression.
]]></description>
<dc:creator>Zhang, J.-X.</dc:creator>
<dc:creator>Smyth, C.</dc:creator>
<dc:creator>Anjum, M. F.</dc:creator>
<dc:creator>Leng, Y.</dc:creator>
<dc:creator>Krystal, A. D.</dc:creator>
<dc:creator>Starr, P. A.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:date>2025-05-13</dc:date>
<dc:identifier>doi:10.1101/2025.05.07.652676</dc:identifier>
<dc:title><![CDATA[A protective role of REM sleep in the pathological basal ganglia-cortical circuit of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.12.653522v1?rss=1">
<title>
<![CDATA[
Conformational biosensors delineate endosomal G protein regulation by GPCRs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.12.653522v1?rss=1</link>
<description><![CDATA[
Many GPCRs trigger a second phase of G protein-coupled signaling from endosomes after initiating signaling from the plasma membrane. This inherently requires receptors to increase the concentration of active-state G proteins on the endosome membrane, but how this is achieved remains incompletely understood. We addressed this question by dissecting the regulation of G protein abundance and activity on endosomes following activation of several Gs-coupled GPCRs-the {beta}2-adrenergic receptor, the VIP-1 receptor, and the adenosine 2b receptor-that are natively co-expressed and differ in their ability to internalize after activation. We first verify GPCR-triggered redistribution of Gs from the plasma membrane to a mixed population of intracellular membranes, including endosomes, that is both reversible after receptor inactivation and triggered irrespective of the ability of the GPCR to internalize. We next show that GPCRs trigger this redistribution process at native expression levels and describe a method, using conformational biosensors, to detect endosomal activation of endogenous Gs. Applying this method, we show that GPCR-mediated production of active-state Gs on endosomes depends on receptor endocytosis, whereas increasing the net amount of Gs on endosomes does not. Our results support a model for Gs regulation on endosomes mediated by two spatially separated receptor coupling events-one at the plasma membrane controlling endosomal Gs abundance and another at endosomes controlling Gs activity. Additionally, our results reveal location-bias in the selectivity of G protein activation on endosomes that is differentially programmed by GPCRs in a receptor-specific manner.
]]></description>
<dc:creator>Wysolmerski, B.</dc:creator>
<dc:creator>Blythe, E. E.</dc:creator>
<dc:creator>von Zastrow, M.</dc:creator>
<dc:date>2025-05-13</dc:date>
<dc:identifier>doi:10.1101/2025.05.12.653522</dc:identifier>
<dc:title><![CDATA[Conformational biosensors delineate endosomal G protein regulation by GPCRs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.09.653201v1?rss=1">
<title>
<![CDATA[
Transcriptomic comparison of early onset preeclampsia and placenta accreta identifies inverse trophoblast and decidua functions at the maternal-fetal interface 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.09.653201v1?rss=1</link>
<description><![CDATA[
Early onset preeclampsia is a placental disorder characterized by shallow implantation, whereas placenta accreta spectrum is a placental disorder of deep placental attachment. This study compares the transcriptome of these two obstetric syndromes. By integrating available microarray and single-cell placenta/decidua transcriptomic datasets, we demonstrated that early onset preeclampsia genes are inversely expressed in placenta accreta, with the most marked differences noted in cell types of decidua, endothelial, and extravillous trophoblasts. Our findings highlight the key functions of trophoblast cell migration and invasion, decidua cell signaling, hypoxia pathways, and global growth factor and collagen contributions to these pregnancy disorders. This research provides new insights into the mechanisms of placentation and unifies these clinical siloes of disease by focusing on the fundamental biology of placental development at the maternal-fetal interface.
]]></description>
<dc:creator>Yin, O.</dc:creator>
<dc:creator>Almonte-Loya, A.</dc:creator>
<dc:creator>Appierdo, R.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Yilmaz, B. D.</dc:creator>
<dc:creator>Oskotsky, T. T.</dc:creator>
<dc:creator>Gonzalez, J. M.</dc:creator>
<dc:creator>Giudice, L. C.</dc:creator>
<dc:creator>Afshar, Y.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.09.653201</dc:identifier>
<dc:title><![CDATA[Transcriptomic comparison of early onset preeclampsia and placenta accreta identifies inverse trophoblast and decidua functions at the maternal-fetal interface]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.13.653825v1?rss=1">
<title>
<![CDATA[
First Generation Tools for the Modeling of Capicua (CIC) - Family Fusion Oncoprotein-Driven Cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.13.653825v1?rss=1</link>
<description><![CDATA[
Clinical divergence between patients harboring CIC-rearrangements is frequently observed. For example, the prototypical CIC::DUX4 fusion associates with soft tissue tumors while CIC::NUTM1 fusions typically localize to the CNS (brain/spinal cord). The basis for these differences is poorly understood due to a lack of molecular tools. To address this need, we generated patient-informed, synthetic coding sequences for CIC::NUTM1, CIC::LEUTX, and ATXN1::DUX4 and validated them in structure-function studies. We found that CIC::NUTM1 drives a transcriptional program distinct from that of CIC::DUX4 due to a C-terminal NUTM1 functional domain, CIC::LEUTX weakly activates CIC target genes through LEUTX transactivation sequences, and ATXN1::DUX4 upregulates CIC target genes via the ATXN1 AXH domain. Our findings indicate that the CIC fusion binding partner may alter overall fusion oncoprotein activity. Thus, these first generation synthetic tools provide an unprecedented resource to study CIC-family fusions beyond CIC::DUX4 and allow for the dissection of this rare subgroup of cancers.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=195 SRC="FIGDIR/small/653825v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Luck, C.</dc:creator>
<dc:creator>Jacobs, K. A.</dc:creator>
<dc:creator>Riad, J.</dc:creator>
<dc:creator>Macaraig, C. D.</dc:creator>
<dc:creator>Ponce, R. K. M.</dc:creator>
<dc:creator>Okimoto, R. A.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.653825</dc:identifier>
<dc:title><![CDATA[First Generation Tools for the Modeling of Capicua (CIC) - Family Fusion Oncoprotein-Driven Cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.09.653155v1?rss=1">
<title>
<![CDATA[
Affinity-matured CD72-targeting Nanobody CAR T-cells Enhance Elimination of Antigen-Low B-cell Malignancies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.09.653155v1?rss=1</link>
<description><![CDATA[
BackgroundChimeric antigen receptor (CAR) T-cell therapies are highly efficacious for several different hematologic cancers. However, for most CAR T targets it is observed that low surface antigen density on tumors can significantly reduce therapeutic efficacy. Here, we explore this dynamic in the context of CD72, a surface antigen we recently found as a promising target for refractory B-cell cancers, but for which CD72 low antigen density can lead to therapeutic resistance in preclinical models.

MethodsPrimary samples were accessed via institutional review board-approved protocols. Affinity-matured and humanized nanobody clones were previously described in Temple et al.1 CAR T-cells were generated via lentiviral transduction. In vitro cytotoxicity assays were performed using luciferase-labeled cell lines. In vivo studies were performed using cell line- or patient-derived xenografts implanted in NOD scid gamma (NSG) mice.

ResultsWe first confirmed ubiquitous CD72 expression across a range of primary B-cell non-Hodgkin lymphomas. We further found that after resistance to CD19-directed therapies, across both B-cell acute lymphoblastic leukemia (B-ALL) models and primary tumor samples, surface CD72 expression was largely preserved while CD22 expression was significantly diminished. Affinity maturation of a nanobody targeting CD72, when incorporated into chimeric antigen receptor (CAR) T-cells, led to more effective elimination in vitro of isogenic models of CD72 low-expressing tumors. These results suggested that nanobody-based CAR T-cells (nanoCARs) may exhibit a similar relationship between binder affinity, antigen expression, and efficacy as previously demonstrated only for scFv-based CAR T-cells. Surprisingly, however, this significantly improved in vitro efficacy only translated to modest in vivo survival benefit. As a parallel strategy to enhance CAR T function, we found that the small molecule bryostatin could also significantly increase CD72 surface antigen density on B-cell malignancy models. Structural modeling and biochemical analysis identified critical residues improving CD72 antigen recognition of our lead affinity-matured nanobody.

ConclusionsTogether, these findings support affinity-matured CD72 nanoCARs as a potential immunotherapy product for CD19-refractory B-cell cancers. Our results also suggest that for B-ALL in particular, CD72 may be a preferable second-line immunotherapy target over CD22.

What is already known on this topicPrevious work using single chain variable fragment (scFv) based CAR Ts has suggested that improving affinity for target antigen could potentially help mitigate tumor resistance mediated by antigen downregulation, or baseline low antigen density. However, it is unknown whether this same dynamic holds for CAR T-cells that utilize different antigen recognition elements, such as nanobodies.

What this study addsHere we show that affinity maturation of nanobody-based CAR T-cells (nanoCARs) targeting CD72 can improve their in vitro efficacy versus CD72-low tumors; however, in vivo efficacy differences are more modest. Furthermore, we show that for refractory B-cell malignancies, surface CD72 appears preserved after CD19 resistance even in situations where CD22 is strongly downregulated.

How this study might affect research, practice or policyCD72 warrants further investigation as a preferred immunotherapy target in the context of CD19-refractory B-cell cancers, though nanobody affinity maturation is not a universal solution to the challenge of low tumor surface antigen density.
]]></description>
<dc:creator>Izgutdina, A.</dc:creator>
<dc:creator>Rashid, T.</dc:creator>
<dc:creator>Temple, W. C.</dc:creator>
<dc:creator>Patino-Escobar, B.</dc:creator>
<dc:creator>Walunj, S.</dc:creator>
<dc:creator>Geng, H.</dc:creator>
<dc:creator>Takamatsu, H.</dc:creator>
<dc:creator>Gil-Alos, D.</dc:creator>
<dc:creator>Kang, A. S.</dc:creator>
<dc:creator>Ramos, E.</dc:creator>
<dc:creator>Chen, S.-Y.</dc:creator>
<dc:creator>Johnson, H.</dc:creator>
<dc:creator>Nix, M. A.</dc:creator>
<dc:creator>Naik, A.</dc:creator>
<dc:creator>Yuan, C. M.</dc:creator>
<dc:creator>Wang, H.-W.</dc:creator>
<dc:creator>Aminov, S.</dc:creator>
<dc:creator>Sahu, S.</dc:creator>
<dc:creator>Larson, R. C.</dc:creator>
<dc:creator>Carpenter, C.</dc:creator>
<dc:creator>Salangsang, F.</dc:creator>
<dc:creator>Phojanakong, P.</dc:creator>
<dc:creator>Camara Serrano, J. A.</dc:creator>
<dc:creator>Tariq, I.</dc:creator>
<dc:creator>Zakraoui, O.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Valeri, A.</dc:creator>
<dc:creator>Martinez-Lopez, J.</dc:creator>
<dc:creator>Maus, M. V.</dc:creator>
<dc:creator>Parekh, S.</dc:creator>
<dc:creator>Verma, A.</dc:creator>
<dc:creator>Shah, N. N.</dc:creator>
<dc:creator>Wiita, A.</dc:creator>
<dc:date>2025-05-15</dc:date>
<dc:identifier>doi:10.1101/2025.05.09.653155</dc:identifier>
<dc:title><![CDATA[Affinity-matured CD72-targeting Nanobody CAR T-cells Enhance Elimination of Antigen-Low B-cell Malignancies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.12.653542v1?rss=1">
<title>
<![CDATA[
GFP-Free Live-Neuron Quantitative Imaging Reveals Compartmentalization and Growth Dynamics of PolyQ Aggregates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.12.653542v1?rss=1</link>
<description><![CDATA[
Huntingtons Disease (HD), the most prevalent polyglutamine (polyQ) neurodegenerative disorder, features brain aggregates induced by mutant huntingtin (mHtt) proteins harboring expanded polyQ tracts. Despite extensive efforts, molecular mechanisms of polyQ aggregates remain elusive. Here we establish quantitative stimulated Raman scattering imaging of polyQ aggregates (q-aggSRS) for non-invasive investigations in live neuronal co-cultures using deuterated glutamine labeling. Q-aggSRS allows for specific visualization by targeting the distinct Raman peak from carbon-deuterium bonds, eliminating the need for bulky GFP tagging. Coupled with analysis from aggregate-tailored expansion microscopy, newly designed two-color imaging, and pulse-chase visualization, we comprehensively quantified the mHtt and non-mHtt proteins within the same aggregates across varying sizes, cell types, mHtt constructs, and subcellular locations. Our findings demonstrate a two-phase aggregate growth model with a distinct core-shell spatial organization, reveal significant heterogeneity in nucleus/cytoplasm compartmentalization specific to neurons, and identify previously unrecognized "gel-like" aggregates specifically in neuronal nuclei. These insights should advance our understanding of native polyQ aggregates and our proposed interaction coefficients may offer new quantitative parameters for developing effective HD therapies.
]]></description>
<dc:creator>Bi, X.</dc:creator>
<dc:creator>Lin, L.-E.</dc:creator>
<dc:creator>Miao, K.</dc:creator>
<dc:creator>Wei, L.</dc:creator>
<dc:date>2025-05-15</dc:date>
<dc:identifier>doi:10.1101/2025.05.12.653542</dc:identifier>
<dc:title><![CDATA[GFP-Free Live-Neuron Quantitative Imaging Reveals Compartmentalization and Growth Dynamics of PolyQ Aggregates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.12.653619v1?rss=1">
<title>
<![CDATA[
Genetic Surveillance Reveals Differential Evolutionary Dynamic of Anopheles gambiae Under Contrasting Insecticidal Tools used in Malaria control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.12.653619v1?rss=1</link>
<description><![CDATA[
Malaria, a febrile disease caused by the Plasmodium parasites and transmitted by mosquitoes, is a leading cause of mortality in children under 5 in endemic countries. The widespread deployment of insecticide-treated bed nets (ITNs) has significantly reduced malaria transmission, but rising levels of insecticide resistance threatens to halt the progress. Monitoring insecticide resistance is vital for effective vector control, particularly when deploying new tools. Understanding mosquito population responses to these interventions is crucial for guiding control programmes in making informed decisions about the selection, timing, and geographic deployment of tools. This genomic study investigates the demographic and evolutionary consequences on the malaria vector Anopheles gambiae of deploying standard ITNs (containing only pyrethroids) and pyrethroid-PBO nets (containing pyrethroids plus the synergist piperonyl butoxide) during a clinical trial in Uganda.

Despite substantial reductions in indoor mosquito densities in the clinical trial, estimates of nucleotide diversity ({pi}) and linkage disequilibrium revealed no significant decline in effective population size, reflecting continued large population size even after effective control. Marked allele frequency shifts at resistance-associated loci indicated strong selection pressures driven by the interventions, with distinct selective dynamics between the two net types, highlighting alternative pyrethroid detoxification pathways in the presence of PBO. A duplication in the Cyp9k1 gene significantly increased in frequency in populations exposed to pyrethroid-only nets but decreased in populations exposed to PBO-treated nets, suggesting that selection for over-expression of this gene is removed when this resistance mechanism is impacted by PBO. An alternative potential detoxification mechanism was selected within a region of the 2La chromosomal inversion on chromosome 2L, which encompasses the UDP-glucose 6-dehydrogenase gene. This variant consistently increased in frequency when exposed to PBO-treated nets. Additionally, pyrethroid-only nets selected for a novel locus on the X chromosome containing the diacylglycerol kinase gene, which is potentially linked to behavioural adaptations through its role in neurotransmission modulation.

Our findings underscore the importance of genomic surveillance in vector control, revealing distinct evolutionary dynamics of insecticide resistance mechanisms in the presence of PBO. While ITNs remain effective, the persistence and evolution of resistance-associated alleles highlight the need for adaptive and dynamic resistance management strategies. By integrating high-resolution genomic data with epidemiological and entomological monitoring, this study offers actionable insights to sustain malaria control efforts amid the ongoing challenge of insecticide resistance.
]]></description>
<dc:creator>Njoroge, H.</dc:creator>
<dc:creator>Namuli, L.</dc:creator>
<dc:creator>Nagi, S. C.</dc:creator>
<dc:creator>Hernandez-Koutoucheva, A.</dc:creator>
<dc:creator>McDermott, D. P.</dc:creator>
<dc:creator>Knight, E.</dc:creator>
<dc:creator>Gonahasa, S.</dc:creator>
<dc:creator>Lynd, A. R.</dc:creator>
<dc:creator>Oruni, A.</dc:creator>
<dc:creator>Maiteki-Sebuguzi, C.</dc:creator>
<dc:creator>Opigo, J.</dc:creator>
<dc:creator>Yeka, A.</dc:creator>
<dc:creator>Katureebe, A.</dc:creator>
<dc:creator>Kyohere, M.</dc:creator>
<dc:creator>Kamya, M.</dc:creator>
<dc:creator>Dorsey, G.</dc:creator>
<dc:creator>hemingway, J.</dc:creator>
<dc:creator>Staedke, S.</dc:creator>
<dc:creator>Clarkson, C. S.</dc:creator>
<dc:creator>Miles, A.</dc:creator>
<dc:creator>Lucas, E. R.</dc:creator>
<dc:creator>Donnelly, M. J.</dc:creator>
<dc:date>2025-05-15</dc:date>
<dc:identifier>doi:10.1101/2025.05.12.653619</dc:identifier>
<dc:title><![CDATA[Genetic Surveillance Reveals Differential Evolutionary Dynamic of Anopheles gambiae Under Contrasting Insecticidal Tools used in Malaria control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.13.651909v1?rss=1">
<title>
<![CDATA[
Select azo compounds post-translationally modulate HTRA1 abundance and activity potentially through interactions at the trimer interface 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.13.651909v1?rss=1</link>
<description><![CDATA[
High-temperature requirement protein A1 (HTRA1) is a secreted serine protease with diverse substrates, including extracellular matrix proteins, proteins involved in amyloid deposition, and growth factors. Accordingly, HTRA1 has been implicated in a variety of neurodegenerative diseases including a leading cause of blindness in the elderly, age-related macular degeneration (AMD). In fact, genome wide association studies have identified that the 10q26 locus which contains HTRA1 confers the strongest genetic risk factor for AMD. A recent study has suggested that AMD-associated risk alleles in HTRA1 correlate with a significant age-related defect in HTRA1 synthesis in the retinal pigmented epithelium (RPE) within the eye, possibly accounting for AMD susceptibility. Thus, we sought to identify small molecule enhancers of HTRA1 transcription and/or protein abundance using an unbiased high-throughput screening approach. To accomplish this goal, we used CRISPR/Sp.Cas9 engineering to introduce an 11 amino acid luminescent peptide tag (HiBiT) onto the C-terminus of HTRA1 in immortalized ARPE-19 cells. Editing was very efficient ([~]88%), verified by genomic DNA analysis, short interfering RNA (siRNA), and HiBiT blotting. Nineteen-hundred and twenty compounds from two libraries were screened. An azo compound with reported anti-amyloidogenic and cardioprotective activity, Chicago Sky Blue 6B (CSB), was identified as an enhancer of endogenous HTRA1 secretion (2.0 {+/-} 0.3 fold) and intracellular levels (1.7 {+/-} 0.2 fold). These results were counter-screened using HiBiT complement factor H (CFH) edited ARPE-19 cells, verified using HiBiT blotting, and were not due to HTRA1 transcriptional changes. Importantly, serine hydrolase activity-based protein profiling (SH-ABPP) demonstrated that CSB does not affect HTRA1s specific activity. However, interestingly, in follow-up studies, Congo Red, another azo compound structurally similar to CSB, also substantially increased intracellular HTRA1 levels (up to 3.6 {+/-} 0.3 fold) but was found to significantly impair HTRA1 enzymatic reactivity (0.45 {+/-} 0.07 fold). Computational modeling of potential azo dye interaction with HTRA1 suggests that CSB and Congo Red can bind to the non-catalytic face of the trimer interface but with different orientation tolerances and interaction energies. These studies identify select azo dyes as HTRA1 chemical probes which may serve as starting points for future HTRA1-centered small molecule therapeutics.
]]></description>
<dc:creator>Hulleman, J.</dc:creator>
<dc:creator>Jeon, S.</dc:creator>
<dc:creator>Bali, S.</dc:creator>
<dc:creator>DiCesare, S.</dc:creator>
<dc:creator>Abbas, A.</dc:creator>
<dc:creator>Daniel, S.</dc:creator>
<dc:creator>Ortega, A.</dc:creator>
<dc:creator>Collier, G.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Bhattacharyaa, A.</dc:creator>
<dc:creator>McCoy, M.</dc:creator>
<dc:creator>Joachimiak, L. A.</dc:creator>
<dc:creator>Posner, B.</dc:creator>
<dc:date>2025-05-16</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.651909</dc:identifier>
<dc:title><![CDATA[Select azo compounds post-translationally modulate HTRA1 abundance and activity potentially through interactions at the trimer interface]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.13.652705v1?rss=1">
<title>
<![CDATA[
Monte Carlo track-structure simulation of the impact of Ultra-Hight Dose Rate and oxygen concentration on the Fenton reaction. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.13.652705v1?rss=1</link>
<description><![CDATA[
BackgroundPreclinical investigations studies have shown that FLASH radiotherapy (FLASH-RT), delivering radiation in ultra-high dose rates (UHDR), preserves healthy tissue and reduces toxicity, all while maintaining an effective tumor response compared to conventional radiotherapy (CONV-RT), the combined biological benefit was termed as "FLASH effect". However, the mechanisms responsible for this effect remain unclear. Research demonstrated that oxygen concentration contributes to the FLASH effect, and it has been hypothesized that Fenton reaction might play a role in the "FLASH effect".

PurposeWe propose to investigate the effect of ultra-high dose rate (UHDR), compared to conventional dose rates (CONV), on the Fenton reaction by studying the radiolysis of Fricke solution. The study will focus on how dose, dose rate, and initial oxygen concentration influence the activation of the Fenton reaction.

Methods and MaterialsTOPAS-nBio version 2.0 was used to simulate the radiolysis of the Fricke system. A cubic water phantom of 3{micro}m side was irradiated by 300MeV protons on one of its edges. For UHDR, a proton field (1.5x1.5{micro}m2) was delivered in a single pulse of 1ns width. The protons were accumulated until reached 5Gy or 10Gy absorbed dose. For CONV, the independent history approach was used to mimic 60Co irradiation. For both dose-rates, oxygen concentrations representative of hypoxic and normoxic tissues (10- 250{micro}M) were simulated. The G-value for oxidant ions G(Fe3+) and {Delta}G-value of Fenton reaction (H2O2 + Fe2+[-&gt;] Fe3++*OH+OH-) were scored. The simulations ended after G(Fe3+) achieved steady-state, and calculated yields were compared with published data.

ResultsFor CONV, G(Fe3+) agreed with ICRU-report 34 data by (0.97{+/-}0.1) %. For UHDR, G(Fe3+) agreed with ICRU data by (1.24{+/-}0.1)% and (0.92{+/-}0.1)% for 5Gy and 10Gy, respectively. Notably, UHDR at 10 Gy reduced the occurrence of Fenton reactions by (1.0{+/-}0.1)% and (11.5{+/-}0.1)% at initial oxygen concentrations of 250 {micro}M and 10 {micro}M, respectively. In consequence, UHDR decreased G(Fe3+) by (1.8{+/-}0.1)% and (12.5{+/-}0.1)% at these oxygen levels. Additionally, increasing the absorbed dose to 15 Gy and 20 Gy at low oxygen (10 {micro}M), UHDR further reduced the {Delta}G-value by (15.7{+/-}0.1)% and (18.6{+/-}0.1)%, respectively. The decrease was driven by intertrack effects present in UHDR pulses and its impact on the scavenging effect that oxygen had over hydrogen radicals.

ConclusionsUHDR reduces the yield of Fe3+ (G(Fe3+)) and significantly impacts Fenton reactions, particularly at low oxygen concentrations, while showing minimal effects at higher oxygen levels. This effect becomes more pronounced at higher dose thresholds, such as 10-20 Gy. This emphasizes the important role of the initial oxygen concentration in UHDR and its influence on the Fenton reaction, a mechanism that may contribute to elucidate the FLASH effect.
]]></description>
<dc:creator>Chaoui, M.</dc:creator>
<dc:creator>Tayalati, Y.</dc:creator>
<dc:creator>Bouhali, O.</dc:creator>
<dc:creator>ramos mendez, j.</dc:creator>
<dc:date>2025-05-16</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.652705</dc:identifier>
<dc:title><![CDATA[Monte Carlo track-structure simulation of the impact of Ultra-Hight Dose Rate and oxygen concentration on the Fenton reaction.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.14.653684v1?rss=1">
<title>
<![CDATA[
Benchmarking cerebellar organoids to model autism spectrum disorder and human brain evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.14.653684v1?rss=1</link>
<description><![CDATA[
While cortical organoids have been used to model different facets of neurodevelopmental conditions and human brain evolution, cerebellar organoids have not yet featured so prominently in the same context, despite increasing evidence of this brain regions importance for cognition and behavior. Here, we provide a longitudinal characterization of cerebellar organoids benchmarked against human fetal data and identify at very early stages of development a significant number of dynamically expressed genes relevant for neurodevelopmental conditions such as autism and attention deficit hyperactivity disorders. Then, we model an ASD mutation impacting CHD8, showing both granule cell and oligodendrocyte lineages prominently affected, resulting in altered network activity in more mature organoids. Lastly, using CRISPR/Cas9 editing, we also model an evolution-relevant mutation in a regulatory region of the CADPS2 gene. We investigate the effect of the derived allele exclusive to Homo sapiens, identifying a rerouting of the CADPS2-expression in rhombic lip cells, coupled with a different sensitivity to hypoxia which in turn lead to a differential timing of granule cell differentiation.

HIGHLIGHTSO_LILongitudinal characterization of cerebellar organoids uncovers disorder related genes especially at early stages of development
C_LIO_LIMutation in CHD8 alter rhombic lip and oligodendrocytes differentiation via WNT pathway
C_LIO_LIRerouting of CADPS2 expression, delaying differentiation and migration, in recent human evolution
C_LI

IN BRIEFAprile and colleagues longitudinally profiled cerebellar organoids, benchmarking them against a fetal human atlas and identified a highly dynamic expression of genes related to cognitive and behavioral disorders especially at early stages of differentiation. Organoids were used to model the impact of a high-penetrance mutation associated with autism spectrum disorder and a high-frequency derived allele in Homo sapiens predicted to have played a role in recent brain evolution.
]]></description>
<dc:creator>Aprile, D.</dc:creator>
<dc:creator>Leonardi, O.</dc:creator>
<dc:creator>Petrella, A.</dc:creator>
<dc:creator>Castaldi, D.</dc:creator>
<dc:creator>Culotta, L.</dc:creator>
<dc:creator>Cheroni, C.</dc:creator>
<dc:creator>Valente, A.</dc:creator>
<dc:creator>Bonfanti, M.</dc:creator>
<dc:creator>Vitriolo, A.</dc:creator>
<dc:creator>Moriano, J.</dc:creator>
<dc:creator>Mirabella, F.</dc:creator>
<dc:creator>Tintori, A.</dc:creator>
<dc:creator>Boeckx, C.</dc:creator>
<dc:creator>Testa, G.</dc:creator>
<dc:date>2025-05-16</dc:date>
<dc:identifier>doi:10.1101/2025.05.14.653684</dc:identifier>
<dc:title><![CDATA[Benchmarking cerebellar organoids to model autism spectrum disorder and human brain evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.15.654128v1?rss=1">
<title>
<![CDATA[
Oxytocin receptor absence reduces selectivity in peer relationships and alters neurochemical release dynamics in prairie voles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.15.654128v1?rss=1</link>
<description><![CDATA[
Abstract/SummaryFriendships, or selective peer relationships, are a vital component of healthy social functioning in humans, while deficits in these relationships are associated with negative physical and mental health consequences. Like humans, prairie voles are among the few mammalian species that form selective social bonds with both peers and mates, making them an excellent model for the mechanistic investigation of selective social attachment. Here, we explored the role of oxytocin receptors in selective peer attachment using prairie voles lacking a functional oxytocin receptor gene (Oxtr1-/-). We found that Oxtr1-/- animals exhibited significant delays in peer relationship formation compared to wildtype animals. Oxytocin receptor function also contributed to the maintenance of peer bonds, as Oxtr1-/- voles displayed reduced relationship stability and lost selective attachments rapidly in a multi-chamber, group-living habitat. Oxtr1-/- voles also showed deficits in both general social reward as well as selective social reward for a peer partner over an unfamiliar conspecific. Evoked oxytocin release in the nucleus accumbens was reduced in Oxtr1-/- animals compared to their wildtype counterparts, indicating that these voles do not have a compensatory increase in oxytocinergic signaling. Together, these data indicate that oxytocin receptors influence the formation, persistence, and reward value of peer relationships.
]]></description>
<dc:creator>Black, A. M.</dc:creator>
<dc:creator>Komatsu, N.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Taskey, S. R.</dc:creator>
<dc:creator>Serrano, N. S.</dc:creator>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Manoli, D. S.</dc:creator>
<dc:creator>Landry, M. P.</dc:creator>
<dc:creator>Beery, A.</dc:creator>
<dc:date>2025-05-16</dc:date>
<dc:identifier>doi:10.1101/2025.05.15.654128</dc:identifier>
<dc:title><![CDATA[Oxytocin receptor absence reduces selectivity in peer relationships and alters neurochemical release dynamics in prairie voles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.17.654540v1?rss=1">
<title>
<![CDATA[
Single Cell RNA Sequencing Reveals Gene Expression Continuums Along the Spatial Hierarchy of the Pulmonary Circulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.17.654540v1?rss=1</link>
<description><![CDATA[
BackgroundThe lung vasculature is comprised of a series of branching vessels extending from the main pulmonary artery to the alveolar capillaries, then back to the pulmonary veins. Lung endothelial cells (EC) exist along this continuum, exposed to gradients of shear stress, oxygen tension and pressure. Single cell RNA sequencing (scRNA-seq) has identified lung EC subsets, but many aspects of the vascular continuum, including vessel size and capillary polarity remain undefined from transcriptomic data.

MethodsWe created an endothelial-enriched scRNA-seq dataset from the P3 mouse lung. Using diffusion pseudotime across all lung EC, we developed an analytical framework to delineate transcriptomic gradients and assign vessel-size scores to categorize individual endothelial cells (EC) along the vascular continuum. We confirmed size-related gene expression patterns with fluorescence in situ hybridization.

ResultsWe categorized capillary 1, arterial and venous EC along two gradients: arterio-venous zonation and vessel size. This approach distinguished large arteries from arterioles, large veins from venules, and revealed arterio-venous polarity within the capillaries. Our data recapitulated previously established zonally defined cell signaling axes, such as high Cxcl12-Cxcr4 signaling in arterioles. We also identified unique cellular communication occurring in large versus small arteries and veins, and localized injury-induced venous EC proliferation to vessels of specific size. This analytical framework was successfully applied to several published mouse and human datasets across different stages of lung development.

ConclusionsThese findings provide a comprehensive transcriptional map of EC across the pulmonary vascular tree, enabling assignment of each individual cell to vessels with defined size and position. This framework offers spatial inferences and novel mechanistic insights from scRNA-seq data sets that may elucidate therapeutic targets to treat pulmonary vascular diseases affecting specific vascular segments. We speculate that similar frameworks could be applied to tissues outside the lung.
]]></description>
<dc:creator>Sveiven, S.</dc:creator>
<dc:creator>Knutsen, C.</dc:creator>
<dc:creator>Zanini, F.</dc:creator>
<dc:creator>Cornfield, D.</dc:creator>
<dc:creator>Alvira, C.</dc:creator>
<dc:date>2025-05-19</dc:date>
<dc:identifier>doi:10.1101/2025.05.17.654540</dc:identifier>
<dc:title><![CDATA[Single Cell RNA Sequencing Reveals Gene Expression Continuums Along the Spatial Hierarchy of the Pulmonary Circulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.19.654776v1?rss=1">
<title>
<![CDATA[
Structural basis for Porcupine inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.19.654776v1?rss=1</link>
<description><![CDATA[
Wnt signalling is essential for embryonic development and tissue homeostasis, and its dysregulation is associated with multiple types of cancer. Porcupine (PORCN), an endoplasmic reticulum (ER)-resident membrane-bound O-acyltransferase, catalyses the palmitoylation of all 19 human Wnts--a critical modification required for their secretion and activity. This central role makes PORCN an attractive therapeutic target for Wnt-driven cancers, with several inhibitors currently in clinical trials. Here, we present high-resolution cryo-electron microscopy structures of human PORCN in complex with the inhibitors C59 (2.4 [A]) and ETC159 (2.6 [A]), as well as in a ligand-free state (3.3 [A]). These structures reveal critical ordered water molecules that form a hydrogen-bonding network within the active site, mediating inhibitor binding. Our docking simulations of diverse PORCN inhibitors demonstrate that despite their different chemical scaffolds, these compounds adopt similar conformations within the acyl-CoA binding site and are also engaged through a conserved water molecule. Our findings provide a structural foundation for the rational design of next-generation PORCN inhibitors with improved pharmacological properties for cancer therapy.
]]></description>
<dc:creator>Black, K. A.</dc:creator>
<dc:creator>Mobbs, J. I.</dc:creator>
<dc:creator>Venugopal, H.</dc:creator>
<dc:creator>Dite, T. A.</dc:creator>
<dc:creator>Leis, A.</dc:creator>
<dc:creator>Wong, L. L.</dc:creator>
<dc:creator>Dagley, L.</dc:creator>
<dc:creator>Thal, D. M.</dc:creator>
<dc:creator>Glukhova, A.</dc:creator>
<dc:date>2025-05-19</dc:date>
<dc:identifier>doi:10.1101/2025.05.19.654776</dc:identifier>
<dc:title><![CDATA[Structural basis for Porcupine inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.19.652834v1?rss=1">
<title>
<![CDATA[
eIF4G2-Mediated Translation Initiation of Histone Modifiers Is Essential for Intestinal Stem Cell Maintenance and Differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.19.652834v1?rss=1</link>
<description><![CDATA[
eIF4G2 (also known as NAT1, p97, or DAP5) is an evolutionally conserved protein homologous to the C-terminal two-thirds portion of eukaryotic translation initiation factor (eIF) 4G. Despite its abundant and ubiquitous expression, the physiological and pathological functions of eIF4G2 are poorly understood. Here, we show that acute loss of eIF4G2 in adult mice results in rapid weight loss with abnormalities in multiple organs, including impaired maintenance and differentiation of intestinal stem cells. Genome-wide ribosome profiling revealed that eIF4G2 is critical for the translation of key histone modification proteins involved in intestinal stemness. Our study underscores the importance of eIF4G2-mediated translation initiation in multicellular organisms.
]]></description>
<dc:creator>Kunitomi, H.</dc:creator>
<dc:creator>Khaine, A. M.</dc:creator>
<dc:creator>Jamee, R.</dc:creator>
<dc:creator>Lancero, M.</dc:creator>
<dc:creator>Raychaudhuri, A.</dc:creator>
<dc:creator>Perli, S. D.</dc:creator>
<dc:creator>Sato, Y.</dc:creator>
<dc:creator>Iwasaki, M.</dc:creator>
<dc:creator>Ruivo, P. R.</dc:creator>
<dc:creator>Tomoda, K.</dc:creator>
<dc:creator>Mito, M.</dc:creator>
<dc:creator>Shichino, Y.</dc:creator>
<dc:creator>Iwasaki, S.</dc:creator>
<dc:creator>Yamanaka, S.</dc:creator>
<dc:date>2025-05-20</dc:date>
<dc:identifier>doi:10.1101/2025.05.19.652834</dc:identifier>
<dc:title><![CDATA[eIF4G2-Mediated Translation Initiation of Histone Modifiers Is Essential for Intestinal Stem Cell Maintenance and Differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.16.654598v1?rss=1">
<title>
<![CDATA[
Repetitive Levodopa Treatment Drives Cell Type-Specific Striatal Adaptations Associated With Progressive Dyskinesia in Parkinsonian Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.16.654598v1?rss=1</link>
<description><![CDATA[
The use of levodopa to manage Parkinsons disease (PD) symptoms leads to levodopa-induced dyskinesia (LID) and other motor fluctuations, which worsen with disease progression and repeated treatment. Aberrant activity of striatal D1- and D2-expressing medium spiny neurons (D1-/D2-MSNs) underlies LID, but the mechanisms driving its progression remain unclear. Using the 6-OHDA mouse model of PD/LID, we combined in vivo and ex vivo recordings to isolate the effect of repeated treatment in LID worsening and other motor fluctuation-related phenotypes. We found that LID worsening is linked to potentiation of levodopa-evoked responses in both D1-/D2-MSNs, independent of changes in dopamine release or MSN intrinsic excitability. Instead, strengthening of glutamatergic synapses onto D1-MSNs emerged as a key driver. Moreover, we found changes in D2-MSN activity that specifically influenced LID duration, potentially contributing to motor fluctuations, which paralleled a reduction in D2R sensitivity. These findings reveal striatal adaptations contributing to worsening of levodopa-related complications.
]]></description>
<dc:creator>Paz, R. M.</dc:creator>
<dc:creator>Ryan, M. B.</dc:creator>
<dc:creator>Marcott, P. F.</dc:creator>
<dc:creator>Girasole, A. E.</dc:creator>
<dc:creator>Faryean, J.</dc:creator>
<dc:creator>Duong, V.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2025-05-21</dc:date>
<dc:identifier>doi:10.1101/2025.05.16.654598</dc:identifier>
<dc:title><![CDATA[Repetitive Levodopa Treatment Drives Cell Type-Specific Striatal Adaptations Associated With Progressive Dyskinesia in Parkinsonian Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.20.654652v1?rss=1">
<title>
<![CDATA[
Synaptic proteins that aggregate and degrade slower with aging accumulate in microglia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.20.654652v1?rss=1</link>
<description><![CDATA[
Neurodegenerative diseases affect 1 in 12 people globally and remain incurable. Central to their pathogenesis is a loss of neuronal protein maintenance and the accumulation of protein aggregates with aging1,2. We engineered bioorthogonal tools3 which allowed us to tag the nascent neuronal proteome and study its turnover with aging, its propensity to aggregate, and its interaction with microglia. We discovered neuronal proteins degraded on average twice as slowly between 4- and 24-month-old mice with individual protein stability differing between brain regions. Further, we describe the aged neuronal  aggregome encompassing 574 proteins, nearly 30% of which showed reduced degradation. The aggregome includes well-known proteins linked to disease as well as a trove of proteins previously not associated with neurodegeneration. Unexpectedly, we found 274 neuronal proteins accumulated in microglia with 65% also displaying reduced degradation and/or aggregation with age. Among these proteins, synaptic proteins were highly enriched, suggesting a cascade of events emanating from impaired synaptic protein turnover and aggregation to the disposal of these proteins, possibly by the engulfment of synapses by microglia. These findings reveal the dramatic loss of neuronal proteome maintenance with aging which could be causal for age-related synapse loss and cognitive decline.
]]></description>
<dc:creator>Guldner, I. H.</dc:creator>
<dc:creator>Wagner, V. P.</dc:creator>
<dc:creator>Moran Losada, P.</dc:creator>
<dc:creator>Shi, S. M.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Meese, B. T.</dc:creator>
<dc:creator>Oh, H.</dc:creator>
<dc:creator>Le Guen, Y.</dc:creator>
<dc:creator>Lu, N.</dc:creator>
<dc:creator>Wong, P. S.</dc:creator>
<dc:creator>To, N.-S.</dc:creator>
<dc:creator>Garceau, D.</dc:creator>
<dc:creator>Guo, Z.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Sasner, M.</dc:creator>
<dc:creator>Keller, A.</dc:creator>
<dc:creator>Yang, A. C.</dc:creator>
<dc:creator>Cheung, T.</dc:creator>
<dc:creator>Wyss-Coray, T.</dc:creator>
<dc:date>2025-05-21</dc:date>
<dc:identifier>doi:10.1101/2025.05.20.654652</dc:identifier>
<dc:title><![CDATA[Synaptic proteins that aggregate and degrade slower with aging accumulate in microglia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.18.654742v1?rss=1">
<title>
<![CDATA[
Cervical Mucus Viscoelasticity and Sperm Velocity are Correlated and Concentration-dependent In Vitro 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.18.654742v1?rss=1</link>
<description><![CDATA[
BackgroundMucus in the endocervix serves as fertility gatekeeper in the reproductive tract through hormonally regulated changes in biophysical properties. Cervical mucus can thicken to prevent ascension of sperm into the upper reproductive tract or thin to permit fertilization. Current reproductive studies of mucus viscoelastic properties rely on subjective visual appraisal of cervical mucus changes. Our goal was to use particle tracking microrheology (PTMR) to objectively assess cervical mucus viscoelastic properties and associate these measurements with in vitro measures of sperm velocity.

MethodsUsing cervical mucus obtained from rhesus macaques (Macaca mulatta) at necropsy, we used to PTMR to measure viscoelasticity (*) under stepwise, serial dilutions. In parallel we measure sperm velocity using custom sperm tracking and analysis workflows.

ResultsWe report that both mucus * and sperm velocity displayed a concentration-dependent behavior, where * increased as mucus concentration increased, and sperm velocity correspondingly decreased. Viscoelasticity and sperm velocity were strongly negatively correlated (p<0.001).

ConclusionsPTMR and sperm tracking in mucus provide quantitative measure of viscoelastic mucus changes. PTMR is potentially a method for quantitatively assessing fertility potential in the cervix that could be applied to both infertility and contraceptives studies.
]]></description>
<dc:creator>Markovetz, M. R.</dc:creator>
<dc:creator>Wei, S.</dc:creator>
<dc:creator>Celluci, C.</dc:creator>
<dc:creator>Roberts, M.</dc:creator>
<dc:creator>Han, L.</dc:creator>
<dc:date>2025-05-23</dc:date>
<dc:identifier>doi:10.1101/2025.05.18.654742</dc:identifier>
<dc:title><![CDATA[Cervical Mucus Viscoelasticity and Sperm Velocity are Correlated and Concentration-dependent In Vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.19.654081v1?rss=1">
<title>
<![CDATA[
Actionable biological programs to enhance EGFR-targeted therapy response unveiled by single-cell lineage tracing in clinically relevant lung cancer models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.19.654081v1?rss=1</link>
<description><![CDATA[
Developing high-resolution approaches to capture both tumor architectural clonality and transcriptional state(s) in individual cells within heterogeneous tumor cell populations could shed light on the evolution of pre-existing and newly emergent tumor subclones and their phenotypes, elucidating their trajectories in response to selective pressures such as drug treatment. Reports to date have focused primarily on analyzing the drug-induced evolution of lung cancer cells in in vitro preclinical models with limited complexity and a relative lack of characterization of actionable biological programs to induce durable responses. Here, we challenged this paradigm and employed a lineage tracing single-cell RNAseq method to track the evolution of primary non-small cell lung cancer (NSCLC) patient-derived organoids (PDOs) and tumor xenografts in response to the standard-of-care EGFR inhibitor osimertinib, with a focus on understanding drug persistence and resistance. Our single-cell lineage tracing-RNAseq system revealed the presence of a discrete set of lineages with distinct transcriptional phenotypes over the course of the treatment. We identified two lineage populations that became predominant during drug treatment and resisted therapy in the PDOs and tumor xenografts. These lineages were present before treatment and harbored Hedgehog pathway and FOXD1 transcriptional programs, respectively. These specific transcriptomic lineages were otherwise undetectable by lower-resolution profiling. Functional studies confirmed the protective role that the baseline expression of the Hedgehog pathway and FOXD1 programs in the lineage tumor cell sub-populations exerts upon targeted therapy. The potential clinical relevance of these regulatory programs was validated by cross-analysis of single-cell transcriptomic data obtained from human NSCLC specimens. Overall, our approach identified pre-existing seeds of resistance before therapy and convergent, adaptive mechanisms supporting tumor residual disease and resistant states. This study highlights the utility of high-resolution tracing of tumor clonal heterogeneity with matched single-cell profiling to reveal occult cell states and molecular mechanisms of therapy resistance and develop counteracting strategies.
]]></description>
<dc:creator>Gini, B.</dc:creator>
<dc:creator>Tamaki, W.</dc:creator>
<dc:creator>Yu, J. X.</dc:creator>
<dc:creator>Barbosa, D.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Pourmoghadam, Y.</dc:creator>
<dc:creator>Allegakoen, P.</dc:creator>
<dc:creator>Kim, D.</dc:creator>
<dc:creator>Miglani, S.</dc:creator>
<dc:creator>Elmes, S.</dc:creator>
<dc:creator>Olivas, V.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Bivona, T. G.</dc:creator>
<dc:date>2025-05-23</dc:date>
<dc:identifier>doi:10.1101/2025.05.19.654081</dc:identifier>
<dc:title><![CDATA[Actionable biological programs to enhance EGFR-targeted therapy response unveiled by single-cell lineage tracing in clinically relevant lung cancer models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.19.654975v1?rss=1">
<title>
<![CDATA[
A catalog of ancient proxies for modern genetic variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.19.654975v1?rss=1</link>
<description><![CDATA[
The ability to observe the genomes of past human populations using ancient DNA provides an extraordinary perspective on many fundamental questions in human genetics, including understanding the evolutionary history of variants that underlie human disease and other phenotypes. However, ancient DNA is often damaged and degraded, yielding low-coverage of most nucleotides. Further, many publicly available genotypes for ancient humans are limited to [~]1.23 million specific loci. Thus, variants of interest often fall outside these specific positions, limiting the ability of ancient DNA to shed light on many loci. Here, we address this challenge by quantifying linkage disequilibrium (LD) between modern variants and ancient genotyped variants (AGVs) to generate a catalog enabling rapid identification of proxy variants. We identified 260,732,675 pairs of AGVs and modern variants with a minimum LD threshold hold of R2 [&ge;] 0.2. Even at R2 [&ge;] 0.9, [&ge;] 60% of common variants were linked to an AGV in non-African ancestry groups, as were 34% of common variants in Africans. We evaluated the accuracy of the genotypes inferred from proxy variants in two high-coverage ancient genomes finding that > 90% of genotypes were correctly predicted, even in a 45,000 year old individual. We also find that AGVs are significantly older than expected and that many likely are evolving neutrally. We integrate these results in a database that researchers can easily query to identify ancient proxy variants if their variant of interest is not directly genotyped in ancient humans.
]]></description>
<dc:creator>Brand, C. M.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:date>2025-05-23</dc:date>
<dc:identifier>doi:10.1101/2025.05.19.654975</dc:identifier>
<dc:title><![CDATA[A catalog of ancient proxies for modern genetic variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.21.655402v1?rss=1">
<title>
<![CDATA[
GABAergic signaling by VIP interneurons gates running-dependent visual recovery in the adult brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.21.655402v1?rss=1</link>
<description><![CDATA[
Experience-dependent plasticity in the adult visual cortex is enhanced by locomotion, a process mediated by vasoactive intestinal peptide (VIP)-expressing interneurons. While VIP interneurons are known to signal through both Gamma-aminobutyric acid (GABA) and VIP peptide, the specific contributions of these pathways during different forms of plasticity remain unclear. Monocular deprivation (MD) in adult mice alters cortical responses, though more slowly and differently than during a critical period in early life. Here, we used two-photon calcium imaging in awake adult mice to dissect the roles of VIP and GABA release from VIP interneurons during adult MD and subsequent binocular recovery. We found comparable level of ocular dominance shifts after MD in mice deficient in either peptidergic or GABA signaling, but disrupting GABA signaling impaired recovery of binocular responses. We also showed that running preferentially enhances contralateral eye responses in binocular primary visual cortex. However, this eye-specific modulation of visual responses by running was altered during recovery from MD and was dependent on VIP signaling pathways. These findings highlight the GABA-mediated inhibition by VIP interneurons as a critical pathway for promoting visual restoration in the adult brain.

Significance StatementUsing longitudinal two-photon imaging in awake adult mice with genetically altered signaling path-ways in VIP interneurons, we demonstrate that GABAergic, but not peptidergic, signaling from VIP interneurons is essential for the recovery of binocular vision following monocular deprivation. We further reveal that locomotion modulates cortical responses in an eye-specific manner, a property dynamically reshaped by plasticity and dependent on VIP interneuron function. These findings identify a discrete inhibitory circuit element that links behavioral state to sensory recovery and highlight GABA release from VIP cells as a potential therapeutic target for restoring visual function in adulthood.
]]></description>
<dc:creator>Lebedeva, A.</dc:creator>
<dc:creator>Kling, F.</dc:creator>
<dc:creator>Rakela, B.</dc:creator>
<dc:creator>Stryker, M. P.</dc:creator>
<dc:creator>Sun, Y. J.</dc:creator>
<dc:date>2025-05-23</dc:date>
<dc:identifier>doi:10.1101/2025.05.21.655402</dc:identifier>
<dc:title><![CDATA[GABAergic signaling by VIP interneurons gates running-dependent visual recovery in the adult brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.20.655097v1?rss=1">
<title>
<![CDATA[
The 1p/19q co-deletion induces targetable and imageable vulnerabilities in glucose metabolism in oligodendrogliomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.20.655097v1?rss=1</link>
<description><![CDATA[
BackgroundThe 1p/19q co-deletion is a hallmark of oligodendrogliomas. The goal of this study was to exploit metabolic vulnerabilities induced by the 1p/19q co-deletion for oligodendroglioma therapy and non-invasive imaging.

MethodsWe used stable isotope tracing, mass spectrometry, and genetic and pharmacological approaches to interrogate [U-13C]-glucose metabolism in patient-derived oligodendroglioma models (SF10417, BT88, BT54, TS603, NCH612). We examined whether tracing [6,6-2H]-glucose metabolism using deuterium metabolic imaging (DMI) provided an early readout of treatment response.

ResultsThe expression of the glycolytic enzyme enolase 1 (ENO1; chromosome 1p36.23) was reduced in patient-derived oligodendroglioma cells and patient biopsies due to the 1p/19q co-deletion and histone hypermethylation. Conversely, ENO2 was upregulated, an effect that was driven by mitogen-activated protein kinase (MAPK) signaling and ERK1-mediated phosphorylation and inactivation of the CIC transcriptional repressor in oligodendrogliomas. Genetic ablation of ENO2 or pharmacological inhibition using POMHEX inhibited proliferation with nanomolar potency but was not cytotoxic to oligodendroglioma cells or tumor xenografts. Mechanistically, ENO2 loss abrogated [U-13C]-glucose metabolism to lactate but shunted glucose towards biosynthesis of serine and purine nucleotides, an effect that was driven by phosphoglycerate dehydrogenase (PHGDH). Importantly, the PHGDH inhibitor D8 was synthetically lethal in combination with POMHEX, and the combination induced tumor regression in vivo. Furthermore, DMI of lactate production from [6,6-2H]-glucose provided an early readout of response to combination therapy that preceded MRI-detectable alterations and reflected extended survival.

ConclusionsWe have identified ENO2 and PHGDH as 1p/19q co-deletion-induced metabolic vulnerabilities in oligodendrogliomas and demonstrated that DMI reports on early response to therapy.

KEY POINTSO_LIThe 1p/19q co-deletion upregulates ENO2 in oligodendrogliomas.
C_LIO_LIENO2 inhibition inhibits glycolysis but upregulates serine and nucleotide biosynthesis via PHGDH.
C_LIO_LICombined inhibition of ENO2 and PHGDH is lethal, an effect that can be visualized by DMI.
C_LI

IMPORTANCE OF THE STUDYOligodendrogliomas are devastating primary brain tumors with long-lasting and life-altering effects on physical and cognitive function. The presence of a 1p/19q co-deletion defines oligodendrogliomas. Here, using clinically relevant patient-derived models and patient tissue, we show that the 1p/19q co-deletion leads to loss of the glycolytic enzyme ENO1 and upregulation of ENO2 in oligodendrogliomas. This provides a unique therapeutic opportunity since most cells rely on ENO1 for glycolysis. Targeting ENO2 using the brain-penetrant inhibitor POMHEX abrogates glycolysis but redirects glucose toward serine and nucleotide biosynthesis, an effect that is driven by PHGDH, the rate-limiting enzyme for serine biosynthesis. Importantly, combined treatment with POMHEX and the PHGDH inhibitor D8 is synthetically lethal in vitro and in vivo. Furthermore, visualizing glucose metabolism using DMI provides an early readout of response to therapy that predicts extended survival in vivo. In summary, we have developed a unique integrated metabolic therapy and imaging approach for oligodendrogliomas.
]]></description>
<dc:creator>Udutha, S.</dc:creator>
<dc:creator>Batsios, G.</dc:creator>
<dc:creator>Taglang, C.</dc:creator>
<dc:creator>Gillespie, A. M.</dc:creator>
<dc:creator>Viswanath, P.</dc:creator>
<dc:date>2025-05-24</dc:date>
<dc:identifier>doi:10.1101/2025.05.20.655097</dc:identifier>
<dc:title><![CDATA[The 1p/19q co-deletion induces targetable and imageable vulnerabilities in glucose metabolism in oligodendrogliomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.21.655369v1?rss=1">
<title>
<![CDATA[
Identification and Characterization of PLUTO-201, a Novel Long Non-Coding RNA Associated with Poor Outcomes in Prostate Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.21.655369v1?rss=1</link>
<description><![CDATA[
Despite extensive investigation, the factors promoting aggressive prostate cancer are poorly understood. By performing a comprehensive analysis of whole-genome transcriptome data to identify differential expression across 1,567 patients with prostate cancer, we now report the identification of a novel lncRNA, Prostate Locus of Uncharacterized Transcript Outlier 201 (PLUTO-201), which is strongly associated with metastasis and poor overall survival in men with prostate cancer. We find that overexpression/knockdown of PLUTO-201 in pre-clinical models of prostate cancer modulates proliferation rates and markers of an aggressive phenotype through regulation of steroid biosynthesis and expression of the MHC class I complex, driving increased growth in androgen-depleted conditions and decreased susceptibility to T cell-mediated cytotoxicity. We further find that the heterogeneous nuclear ribonucleoprotein hnRNPK directly binds PLUTO-201 and is indispensable for its activity. Overall, our findings indicate that PLUTO-201 is a driver of aggressive prostate cancer phenotypes and poor clinical outcomes.

Statement of SignificanceIdentification and characterization of PLUTO-201, a novel lncRNA driving aggressive biology in prostate cancer, sheds new light on the mechanisms driving aggressive prostate cancer and will motivate therapeutic and biomarker development.

Statement of Translational RelevanceThe factors promoting prostate cancer progression and metastasis are poorly understood, resulting in a lack of therapeutic targets and prognostic biomarkers for this disease. Here, we have identified the novel long non-coding RNA (lncRNA) PLUTO-201 as strongly associated with prostate cancer progression and metastasis in patients with localized prostate cancer undergoing prostatectomy. We show that PLUTO-201 promotes proliferation, invasion, and metastasis in multiple prostate cancer models both in vitro and in vivo. Mechanistically, we find that PLUTO-201 downregulates MHC class 1 and upregulates steroid biosynthesis by interacting with the heterogeneous nuclear ribonucleoprotein K (hnRNPK), leading to decreased T cell-mediated cytotoxicity and increased resistance to androgen receptor inhibition. Altogether, this study provides strong evidence for a critical role of PLUTO-201 in prostate cancer progression and metastasis, and a rationale for further exploration of PLUTO-201 as a therapeutic target and prognostic biomarker for patients with prostate cancer.
]]></description>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Younger, N. S.</dc:creator>
<dc:creator>Malik, B.</dc:creator>
<dc:creator>Shin, H. J.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Niknafs, Y.</dc:creator>
<dc:creator>Zhao, S.</dc:creator>
<dc:creator>Wilder-Romans, K.</dc:creator>
<dc:creator>Pitchiaya, S.</dc:creator>
<dc:creator>Han, S.</dc:creator>
<dc:creator>Barnard, T. J.</dc:creator>
<dc:creator>Lloyd, P.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Chesner, L. N.</dc:creator>
<dc:creator>Calvert, M.</dc:creator>
<dc:creator>Egusa, E. A.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Chou, J.</dc:creator>
<dc:creator>Das, R.</dc:creator>
<dc:creator>Kothari, V.</dc:creator>
<dc:creator>Shenoy, T.</dc:creator>
<dc:creator>Diolaiti, M. E.</dc:creator>
<dc:creator>Malik, R.</dc:creator>
<dc:creator>Prensner, J. R.</dc:creator>
<dc:creator>Burlingame, A. L.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Chinnaiyan, A. M.</dc:creator>
<dc:creator>Feng, F.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:date>2025-05-24</dc:date>
<dc:identifier>doi:10.1101/2025.05.21.655369</dc:identifier>
<dc:title><![CDATA[Identification and Characterization of PLUTO-201, a Novel Long Non-Coding RNA Associated with Poor Outcomes in Prostate Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.23.655626v1?rss=1">
<title>
<![CDATA[
Aperiodic neural dynamics define a novel signature of glioma-induced excitation-inhibition dysregulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.23.655626v1?rss=1</link>
<description><![CDATA[
Diffuse gliomas remodel neuronal circuits with prognostic and therapeutic significance for patients. Electrophysiologic measures of cortical excitability hold promise for monitoring disease progression and evaluating therapeutic responses. The power law exponent (aperiodic slope) reflects the balance between excitatory and inhibitory activity within neuronal networks, a critical aspect of normal brain function often disrupted in neurological conditions. Despite its potential, the significance of the aperiodic slope in glioma-infiltrated tissue and its underlying cellular processes has not been fully investigated. Here, we integrate multi-modal electrophysiological analysis with transcriptomic profiling to analyze the aperiodic slope in both normal and glioma-infiltrated cortex. We determine that glioma infiltration induces a flattening of the aperiodic slope, indicating a shift toward excitation dominance that varies according to tumor subtype and correlates with impairments in semantic naming. Single-nucleus RNA sequencing revealed that cortical regions with flat aperiodic slope exhibit transcriptional programs enriched in glutamatergic signaling, membrane depolarization, and excitatory synaptic transmission. The aperiodic slope responds to pharmacologically induced changes in cortical inhibition during propofol administration, a GABAA agonist. Our results establish the aperiodic slope as a robust biomarker of glioma-associated excitation-inhibition imbalance, with potential applications in tumor classification and treatment monitoring.
]]></description>
<dc:creator>Sibih, Y. E.</dc:creator>
<dc:creator>Dada, A. O.</dc:creator>
<dc:creator>Cunningham, E. E.</dc:creator>
<dc:creator>Olshausen, N.</dc:creator>
<dc:creator>Kaur, J.</dc:creator>
<dc:creator>Jayabal, V.</dc:creator>
<dc:creator>Oten, S.</dc:creator>
<dc:creator>Herr, S.</dc:creator>
<dc:creator>Gonzales, C. N.</dc:creator>
<dc:creator>Daniel, A.</dc:creator>
<dc:creator>Krishna, S.</dc:creator>
<dc:creator>Ambati, V. S.</dc:creator>
<dc:creator>Aabedi, A. A.</dc:creator>
<dc:creator>Umbach, G.</dc:creator>
<dc:creator>Mirchia, K.</dc:creator>
<dc:creator>Lalwani, P.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Brang, D.</dc:creator>
<dc:creator>Hervey-Jumper, S. L.</dc:creator>
<dc:date>2025-05-24</dc:date>
<dc:identifier>doi:10.1101/2025.05.23.655626</dc:identifier>
<dc:title><![CDATA[Aperiodic neural dynamics define a novel signature of glioma-induced excitation-inhibition dysregulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.20.655215v1?rss=1">
<title>
<![CDATA[
Perivascular matrix densification dysregulates angiogenesis and activates pro-inflammatory endothelial cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.20.655215v1?rss=1</link>
<description><![CDATA[
Fibrosis is central to numerous fatal conditions including solid cancers, pulmonary fibrosis, cirrhosis and post-infarct cardiac fibrosis amongst many others, thereby collectively contributing to 45% of all deaths in developed nations. The potential for fibrosis across most organ systems may stem from its connections to wound healing and the ubiquitous presence of vascular endothelium. Endothelial cells (ECs) and angiogenesis, cells and associated biological program central to wound healing, have been heavily implicated in many organ-specific fibroses, but the relationship between angiogenesis and fibrogenesis remains debated and little has been established in terms of how the EC phenotype governs tissue healing vs. fibrosis. Here, we examine a murine lung injury model enabling EC lineage tracing and observe the invasion of aberrant ECs from the bronchial microvasculature following lung injury along with concurrent densification of matrix fibers surrounding these vessels. To investigate the underlying mechanisms governing their appearance, we established a microphysiological system (MPS) of arteriole/venule-scale microvessels embedded within a tunable stromal mimetic matrix and find that heightened extracellular matrix fiber density activates ECs, drives endothelial to mesenchymal transition, and promotes aberrant tip EC (ATEC) invasion into the matrix. ATECs remain adherent to fibrotic matrix and possess a pro-inflammatory phenotype that secretes TGF-{beta}2. Notably, our studies establish that the formation of ATECs is gated by destabilization of endothelial adherens junction upon EC adhesion to fibrous matrix, and associated regulation of TGF-{beta} signaling that is mediated by a novel VE-cadherin - TGF-{beta}R2 axis. The current lack of effective anti-fibrotic therapies suggests potential critical involvement of other cell types such as ECs, and our findings suggest new contributions of ECs to fibrotic progression that may better inform future targets for novel anti-fibrotic therapeutics.
]]></description>
<dc:creator>Xia, J.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Jacobs, K.</dc:creator>
<dc:creator>Lin, D.</dc:creator>
<dc:creator>Jarman, E.</dc:creator>
<dc:creator>Matera, D.</dc:creator>
<dc:creator>Loesel, K.</dc:creator>
<dc:creator>Davison, C.</dc:creator>
<dc:creator>Hiraki, H.</dc:creator>
<dc:creator>Tan, X.</dc:creator>
<dc:creator>Shikanov, E.</dc:creator>
<dc:creator>Kent, R.</dc:creator>
<dc:creator>Parent, C.</dc:creator>
<dc:creator>Fan, X.</dc:creator>
<dc:creator>Shikanov, A.</dc:creator>
<dc:creator>Kutys, M. L.</dc:creator>
<dc:creator>Baker, B.</dc:creator>
<dc:date>2025-05-25</dc:date>
<dc:identifier>doi:10.1101/2025.05.20.655215</dc:identifier>
<dc:title><![CDATA[Perivascular matrix densification dysregulates angiogenesis and activates pro-inflammatory endothelial cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.23.655442v1?rss=1">
<title>
<![CDATA[
Mapping early patterning events in human neural development usingan in-vitro microfluidic stem cell model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.23.655442v1?rss=1</link>
<description><![CDATA[
Stem cell models can provide insights into human brain development at embryonic stages which are normally inaccessible. We previously developed the Microfluidic Stem Cell Regionalisation (MiSTR) model, which recapitulates the rostro-caudal patterning of human neural tube through a WNT activation (WNTa) gradient. Through temporal single cell transcriptomics of rostro-caudal and dorso-ventral gradient-patterned MiSTR, we found that rostro-caudal subtypes were regionally specified and fate-determined already during the late epiblast stage, several days before onset of neuralisation at day 3-4. Rostral cells were characterised by expression of HESX1 and SHISA2 during pre-neuralisation and PAX6 during early neuralisation, whereas caudal cells expressed FST and HOXA1 during pre-neuralisation and SOX1 as the dominant neuralising factor. In contrast to the early rostro-caudal specification, response to ventralisation in telencephalic progenitors was developmentally delayed and occurred around day 9. We further uncovered temporal events in human midbrain-hindbrain boundary formation and ventral forebrain patterning, contributing new knowledge on early human neural region-specification.
]]></description>
<dc:creator>Rathore, G. S.</dc:creator>
<dc:creator>Ankjaer, M.</dc:creator>
<dc:creator>Rifes, P.</dc:creator>
<dc:creator>Hanninen, E.</dc:creator>
<dc:creator>Calleja, F. S.</dc:creator>
<dc:creator>Rusimbi, C.</dc:creator>
<dc:creator>Kajtez, J.</dc:creator>
<dc:creator>Nygaard, A. H.</dc:creator>
<dc:creator>Piilgaard, L. S.</dc:creator>
<dc:creator>Dubonyte, U.</dc:creator>
<dc:creator>Norgaard, Z. A.</dc:creator>
<dc:creator>Christensen, J. B.</dc:creator>
<dc:creator>Egerod, K. L.</dc:creator>
<dc:creator>Camp, J. G.</dc:creator>
<dc:creator>Pers, T.</dc:creator>
<dc:creator>Treutlein, B.</dc:creator>
<dc:creator>Kirkeby, A.</dc:creator>
<dc:date>2025-05-26</dc:date>
<dc:identifier>doi:10.1101/2025.05.23.655442</dc:identifier>
<dc:title><![CDATA[Mapping early patterning events in human neural development usingan in-vitro microfluidic stem cell model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.21.655323v1?rss=1">
<title>
<![CDATA[
Molecular mechanism of substrate transport by human peroxisomal ABCD3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.21.655323v1?rss=1</link>
<description><![CDATA[
Peroxisomes are eukaryotic oxidative organelles involved in numerous metabolic functions that include fatty acid oxidation, bile acid synthesis, and detoxification of reactive oxygen species. ATP-binding cassette transporters of the D subfamily (ABCD1-3) mediate the import of CoA thioesters of fatty acids into the peroxisome. ABCD3, the most abundant of these transporters in the peroxisomal membrane, facilitates the transport of a broad spectrum of substrates including branched-chain fatty acids, very long-chain fatty acids, bile salt intermediates, and dicarboxylic acids. Mutations in ABCD3 are associated with defects in congenital bile acid synthesis and variants of Zellweger syndrome. The structural and functional details of the human ABCD3 transporter remain unclear, despite its significance. In this study, we report the cryogenic sample electron microscopy (cryo-EM) structures of full-length human ABCD3 in its apo state and bound to one of the physiological substrates (phytanoyl-CoA) at resolutions of 3.33 [A] and 3.13 [A], respectively. Our biochemical assays reveal that substrate binding induces ATPase activity in ABCD3, suggesting a substrate-dependent conformational change. Structural comparison of the apo and substrate bound states demonstrate that the substrate interaction brings nucleotide-binding domains closer, providing a mechanistic basis of substrate induced ATPase activity. These findings offer critical insights into the transport mechanism of ABCD3 and lay a structural foundation for understanding its role in peroxisomal metabolite import and related diseases.

Significance StatementPeroxisomes are involved in essential cellular metabolic processes that include fatty acid oxidation, bile acid synthesis, and detoxification. The ABCD subfamily of membrane transporter proteins transport fatty acyl-CoA molecules from the cytosol into peroxisomes. Among the ABCD subfamily members, ABCD3 transports branched chain fatty acids and a range of other important metabolites. Dysfunction in ABCD3 has been associated with several inherited and acquired diseases. Using cryo-EM, we determined high-resolution structures of full-length human ABCD3 in both unbound and phytanoyl-CoA-bound forms. We show that substrate binding stimulates ATP hydrolysis activity by apposition of the two nucleotide binding domains of ABCD3, adding structural insight into the mechanism of action of ABCD3 and its role in peroxisomal function and disease.
]]></description>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Khandelwal, N. K.</dc:creator>
<dc:creator>Seka, D. J.</dc:creator>
<dc:creator>Balasubramani, S. G.</dc:creator>
<dc:creator>Dickinson, M. S.</dc:creator>
<dc:creator>Myasnikov, A.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Stroud, R. M.</dc:creator>
<dc:date>2025-05-27</dc:date>
<dc:identifier>doi:10.1101/2025.05.21.655323</dc:identifier>
<dc:title><![CDATA[Molecular mechanism of substrate transport by human peroxisomal ABCD3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.22.655511v1?rss=1">
<title>
<![CDATA[
An information theory framework for capturing multi-connectivity via spatial network encoding reveals reduced population count (Hamming weight) localized to auditory, visual, and motor networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.22.655511v1?rss=1</link>
<description><![CDATA[
1.The human brain operates as a complex system where functional networks evolve and interact across spatially distributed regions. In traditional neuroimaging analyses, functional connectivity (FC), based on pairwise correlations or statistical dependencies of temporal fluctuations in the BOLD signal, has been a primary method for exploring interactions between brain regions and decoding brain function. However, traditional FC methods often overlook the intricate, multi-way interplay among brain elements that emerge from the brains densely interconnected nature. To overcome these limitations, we introduce a novel voxel-centric framework that captures the multi-way interactions between voxels and networks identified via high-model order independent component analysis. This framework posits that individual voxels serve as critical mediators of multi-network communication, reflecting the brains complex functional architecture. By encoding voxel contributions from brain networks into binary representations and quantifying the population count at each voxel via Hamming weights, the proposed method prioritizes high-contribution voxels that facilitate inter-network interactions. This approach provides new insights into the brains functional organization, revealing previously unrecognized patterns of voxel-to-network entanglement. Specifically, in the context of schizophrenia, our method enables the identification of spatial patterns that may underpin the cognitive and perceptual disturbances characteristic of the disorder. This enhanced understanding could improve diagnostic precision and help tailor interventions that target specific dysfunctional networks, offering a pathway to more effective treatments and better patient outcomes in schizophrenia.
]]></description>
<dc:creator>Bostami, B.</dc:creator>
<dc:creator>Lewis, N.</dc:creator>
<dc:creator>Agcaoglu, O.</dc:creator>
<dc:creator>Silva, R. F.</dc:creator>
<dc:creator>Turner, J. A.</dc:creator>
<dc:creator>van Erp, T.</dc:creator>
<dc:creator>Ford, J.</dc:creator>
<dc:creator>Calhoun, V.</dc:creator>
<dc:date>2025-05-27</dc:date>
<dc:identifier>doi:10.1101/2025.05.22.655511</dc:identifier>
<dc:title><![CDATA[An information theory framework for capturing multi-connectivity via spatial network encoding reveals reduced population count (Hamming weight) localized to auditory, visual, and motor networks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.22.655585v1?rss=1">
<title>
<![CDATA[
Chondrocyte-Specific Knockout of Piezo1 and Piezo2 Protects Against Post-Traumatic Osteoarthritis Structural Damage and Pain in Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.22.655585v1?rss=1</link>
<description><![CDATA[
AbstractO_ST_ABSBackgroundC_ST_ABSOsteoarthritis (OA) is a debilitating joint disease characterized by cartilage degeneration, synovial inflammation, and bone remodeling, with limited therapeutic options targeting the underlying pathophysiology. Mechanosensitive ion channels Piezo1 and Piezo2 play crucial roles in chondrocyte responses to mechanical stress, mediating mechanotransduction pathways that influence chondrocyte survival, matrix production, and inflammatory signaling, but their distinct contributions to OA pathogenesis remain unclear.

MethodsUsing inducible, chondrocyte-specific Aggrecan-Cre (Acan) mice, we investigated Piezo1, Piezo2, and combined Piezo1/2 conditional knockouts (cKOs) using the destabilization of the medial meniscus (DMM) model of post-traumatic OA in male and female mice. Pain and behavioral assessments were conducted at four time points to evaluate OA progression, while cartilage damage, bone remodeling, and synovial inflammation were assessed at the final endpoint of 28 weeks. Statistical analyses included one-way and two-way ANOVA with Tukeys multiple comparisons test.

ResultsPiezo1 cKO delayed pain onset but ultimately exacerbated cartilage degradation and synovitis, emphasizing its dual role in protective and pathogenic mechanotransduction. While the Piezo2 cKO reduced pain and preserved activity, it failed to protect cartilage. Notably, Piezo1/2 cKO provided the greatest protection against cartilage degeneration, synovitis, and pain. Micro-computed tomography analyses revealed that Piezo2 is critical for maintaining trabecular bone integrity, with a Piezo2 cKO leading to decreased bone volume, thickness, and density, independent of injury. Piezo2 cKO also reduced normal meniscal ossification that occurs with age in mice. In contrast, a Piezo1/2 cKO normalized most bone remodeling parameters observed in Piezo2 cKO mice but did not restore medial tibial plateau thickness, highlighting Piezo2s essential role in bone structure.

ConclusionsThese findings demonstrate the overlapping and compensatory roles of Piezo1 and Piezo2 in OA pathogenesis. Dual inhibition of Piezo1 and Piezo2 may offer a novel, effective therapeutic strategy targeting both structural and symptomatic aspects of the disease.
]]></description>
<dc:creator>Ely, E.</dc:creator>
<dc:creator>Lenz, K.</dc:creator>
<dc:creator>Paradi, S.</dc:creator>
<dc:creator>Ack, S.</dc:creator>
<dc:creator>Behrmann, A.</dc:creator>
<dc:creator>Dunivan, S.</dc:creator>
<dc:creator>Braxton, L.</dc:creator>
<dc:creator>Liedtke, W.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Collins, K. H.</dc:creator>
<dc:creator>Guilak, F.</dc:creator>
<dc:date>2025-05-27</dc:date>
<dc:identifier>doi:10.1101/2025.05.22.655585</dc:identifier>
<dc:title><![CDATA[Chondrocyte-Specific Knockout of Piezo1 and Piezo2 Protects Against Post-Traumatic Osteoarthritis Structural Damage and Pain in Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.22.655605v1?rss=1">
<title>
<![CDATA[
Extracellular Ca2+-Sensing Receptor (CaSR) Regulates Hypothalamic Function to Control Energy and Skeletal Metabolism in Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.22.655605v1?rss=1</link>
<description><![CDATA[
Introductory paragraphThe extracellular Calcium-sensing receptor (CaSR) regulates cellular responsiveness to physiological changes in ionized calcium (Ca2+) concentrations. The CaSR is expressed in the brain, including in hypothalamic growth hormone stimulating GHRH and anorectic POMC neurons that control growth and energy homeostasis. We embryonically deleted the Casr gene in neurons to create NeuronCaSR-/- mice to delineate the role of this receptor in regulating growth, skeletal development, and energy metabolism. NeuronCaSR-/- mice had reduced size, weight and bone mass compared to littermate controls, with a dysregulated growth hormone axis. They also showed increased adiposity and circulating leptin levels, leptin resistance, and decreased glucose tolerance, along with reduced expression of the anorectic precursor peptide POMC and secondary increases in the expression of the anorectic peptide AgRP in the hypothalamus of NeuronCaSR-/- mice. Knockdown of CaSR in adult mice specifically in the hypothalamic arcuate nucleus, where GHRH, POMC and AgRP neurons reside, also resulted in increased body weight, adiposity, leptin resistance, and glucose intolerance, and reduced bone mass. Together these data suggest that neuronal CaSR critically regulates energy and skeletal metabolism and body growth by modulating hypothalamic function, representing a new paradigm for central integration of calcaemic activities with body function.
]]></description>
<dc:creator>Park-sigal, J. J.</dc:creator>
<dc:creator>Norton, M.</dc:creator>
<dc:creator>Tu, C.-L.</dc:creator>
<dc:creator>Cheng, Z.</dc:creator>
<dc:creator>Fadahunsi, N.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Bewick, G. A.</dc:creator>
<dc:creator>Murphy, K. G.</dc:creator>
<dc:creator>Chang, W. A.</dc:creator>
<dc:date>2025-05-27</dc:date>
<dc:identifier>doi:10.1101/2025.05.22.655605</dc:identifier>
<dc:title><![CDATA[Extracellular Ca2+-Sensing Receptor (CaSR) Regulates Hypothalamic Function to Control Energy and Skeletal Metabolism in Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.23.654927v1?rss=1">
<title>
<![CDATA[
A Cellular Basis for Heightened Gut Sensitivity in Females 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.23.654927v1?rss=1</link>
<description><![CDATA[
Visceral pain disorders, such as irritable bowel syndrome, exhibit a marked female prevalence. Enhanced signaling between enterochromaffin (EC) cells in the gut epithelium and mucosal sensory nerve fibers likely contributes to this sex bias. Here, we identify a novel estrogen-responsive paracrine pathway in which two enteroendocrine cell types, PYY-expressing L-cells and serotonergic EC cells, communicate to increase gut sensitivity in females. We demonstrate that ER estrogen signaling upregulates the bacterial metabolite SCFA receptor Olfr78 on colonic L-cells, increasing PYY release and their sensitivity to acetate. Elevated PYY acts on neighboring EC cells via NPY1R, thereby enhancing serotonin release and gut pain. We propose that hormonal fluctuations, in conjunction with internal (stress) or environmental (diet) factors, amplify this local estrogen-responsive colonic circuit, resulting in maladaptive gut sensitivity.
]]></description>
<dc:creator>Venkataraman, A.</dc:creator>
<dc:creator>Figueroa, E. E.</dc:creator>
<dc:creator>Castro, J.</dc:creator>
<dc:creator>Navarro, F. M. C.</dc:creator>
<dc:creator>Soota, D.</dc:creator>
<dc:creator>Brierley, S. M.</dc:creator>
<dc:creator>Julius, D.</dc:creator>
<dc:creator>Ingraham, H. A.</dc:creator>
<dc:date>2025-05-28</dc:date>
<dc:identifier>doi:10.1101/2025.05.23.654927</dc:identifier>
<dc:title><![CDATA[A Cellular Basis for Heightened Gut Sensitivity in Females]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.24.655953v1?rss=1">
<title>
<![CDATA[
Multivalent Lipid MVL5 Micellar Nanoparticles Exhibit Dramatically Increased Loading of Paclitaxel with PEGylation Enhancing Human Cancer Cell Penetration Depth and Cytotoxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.24.655953v1?rss=1</link>
<description><![CDATA[
Cationic liposomes (CLs) with chain-melted fluid membranes are promising nanocarriers of hydrophobic drugs in cancer chemotherapy, including the frequently employed drug paclitaxel (PTX). CL formulations containing univalent N-[2,3-dioleoyloxy-1-propyl]trimethylammonium chloride (DOTAP), like EndoTAG-1TM, have shown limited success in clinical trials and challenges like endosomal entrapment, limited PTX membrane solubility, and difficulty with in vivo tumor targeting remain. Incorporation of 10 mol% cone-shaped poly(ethylene glycol)-lipid (PEG-lipid) to DOTAP-containing CLs transitions a fraction of the particles to micellar nanodiscs. These PTX--loaded PEGylated CLs and nanodiscs show enhanced cellular uptake in vitro and improved tumor penetration and proapoptotic activity compared to bare CLs in an in vivo solid breast cancer tumor model. Formulations incorporating the multivalent cationic lipid MVL5 (+5e) at 50 mol% form nanoparticles (NPs) comprised almost entirely of nanodiscs, and transition at 75 mol% MVL5 to short micellar rods coexisting with spheres, with rods further transitioning to long flexible rods upon PEGylation. Here, we report on the finding that MVL5-based micellar NPs with disc, rod, and spherical morphologies dramatically improve the solubility of PTX in their fluid lipid membranes by nearly three-fold compared to reference CLs modeled on the EndoTAG-1TM formulation. Cell viability assays revealed that this improved PTX solubility for MVL5 micellar NPs leads to improved cytotoxic efficacy, which is further improved by PEGylation. Remarkably, using fluorescent microscopy and image particle analysis, we find that the cellular uptake and penetration depth of MVL5 nanoparticles is significantly improved by PEGylation. The findings are consistent with a model where the rate-limiting step of PTX delivery by cationic lipid NPs is diffusion of endocytic vesicles containing NPs through the actin mesh near the cell surface combined with the hoping rate of PTX from endosomal membrane to nearby microtubules. PEGylated cationic lipid nanoparticles containing MVL5 therefore represent a very promising hydrophobic cancer drug delivery vehicle for nanomedicine applications.
]]></description>
<dc:creator>Fisher, W. S.</dc:creator>
<dc:creator>Ghasemizadeh, A.</dc:creator>
<dc:creator>Roshan, S.</dc:creator>
<dc:creator>Goldstein, A.</dc:creator>
<dc:creator>Douglas, J.</dc:creator>
<dc:creator>Perez, R.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Ewert, K. K.</dc:creator>
<dc:creator>Safinya, C. R.</dc:creator>
<dc:date>2025-05-29</dc:date>
<dc:identifier>doi:10.1101/2025.05.24.655953</dc:identifier>
<dc:title><![CDATA[Multivalent Lipid MVL5 Micellar Nanoparticles Exhibit Dramatically Increased Loading of Paclitaxel with PEGylation Enhancing Human Cancer Cell Penetration Depth and Cytotoxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.27.656446v1?rss=1">
<title>
<![CDATA[
Caffeine Impairs Red Blood Cell Storage Quality by Dual Inhibition of ADORA2b Signaling and G6PD Activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.27.656446v1?rss=1</link>
<description><![CDATA[
Caffeine is the most widely consumed psychoactive substance globally, yet its peripheral physiological effects remain incompletely understood. Leveraging comprehensive data from 13,091 blood donors in the REDS RBC-Omics study, we identify caffeine as a significant modulator of red blood cell (RBC) storage quality and transfusion outcomes. Elevated caffeine levels were reproducible across multiple donations from 643 recalled donors, selected based on their extremes in hemolytic propensity. Both in the screening and recalled cohorts, higher caffeine levels were associated with disrupted RBC metabolism, characterized by reduced glycolysis, depletion of adenylate pools or 2,3-bisphosphoglycerate, and increased markers of oxidative stress and osmotic fragility, including kynurenine accumulation. These observations were recapitulated in plasma and RBCs of eight volunteers upon consumption of a cup of coffee independently of brewing method (Chemex vs espresso). Clinically, elevated caffeine correlated with increased hemolysis and lower post-transfusion hemoglobin increments, especially pronounced in recipients transfused with RBCs from donors carrying common polymorphisms in the ADORA2b gene, a key regulator of RBC metabolism in hypoxia. These human findings were mechanistically validated using a murine model deficient in ADORA2b, which demonstrated impaired glycolytic flux, compromised antioxidant defenses - including caffeine-dependent direct inhibition of recombinantly-expressed glucose 6-phosphate dehydrogenase, and decreased transfusion efficacy (lower hemoglobin increments, higher bilirubin post-transfusion), effects further exacerbated by caffeine exposure during storage. Our study positions caffeine consumption as a modifiable factor in blood transfusion practice, advocating for precision strategies that integrate genetic and exposome factors, and identifies metabolic interventions to enhance blood quality and clinical outcomes.

One sentence summaryCaffeine consumption and genetic variants in the ADORA2b receptor synergistically impair red blood cell metabolism and transfusion efficacy, revealing a modifiable exposome-gene interaction for precision transfusion medicine.
]]></description>
<dc:creator>Dzieciatkowska, M.</dc:creator>
<dc:creator>Hay, A.</dc:creator>
<dc:creator>Issaian, A.</dc:creator>
<dc:creator>Keele, G. R.</dc:creator>
<dc:creator>Bevers, S.</dc:creator>
<dc:creator>Nemkov, T.</dc:creator>
<dc:creator>Reisz, J. A.</dc:creator>
<dc:creator>Maslanka, M.</dc:creator>
<dc:creator>Stephenson, D.</dc:creator>
<dc:creator>Moore, A.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>Kleinman, S.</dc:creator>
<dc:creator>Norris, P. J.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>Page, G. P.</dc:creator>
<dc:creator>Roubinian, N. H.</dc:creator>
<dc:creator>Xia, Y.</dc:creator>
<dc:creator>Zimring, J. C.</dc:creator>
<dc:creator>D'Alessandro, A.</dc:creator>
<dc:date>2025-05-30</dc:date>
<dc:identifier>doi:10.1101/2025.05.27.656446</dc:identifier>
<dc:title><![CDATA[Caffeine Impairs Red Blood Cell Storage Quality by Dual Inhibition of ADORA2b Signaling and G6PD Activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.28.656516v1?rss=1">
<title>
<![CDATA[
A non-spike nucleocapsid R204P mutation in SARS-CoV-2 Omicron XEC enhances inflammation and pathogenicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.28.656516v1?rss=1</link>
<description><![CDATA[
The global circulation of SARS-CoV-2 in human populations has driven the emergence of Omicron subvariants, which have become highly diversified through recombination. In late 2024, SARS-CoV-2 Omicron XEC variant emerged from the recombination of two JN.1 progeny, KS.1.1 and KP.3.3, and became predominant worldwide. Here, we investigated virological features of the XEC variant. Epidemic dynamics modeling suggested that spike substitutions in XEC mainly contribute to its increased viral fitness. Additionally, four licensed antivirals were effective against XEC. Although the fusogenicity of XEC spike is comparable to that of the JN.1 spike, the intrinsic pathogenicity of XEC in hamsters was significantly higher than that of JN.1. Notably, we found that the nucleocapsid R204P mutation of XEC enhanced inflammation through NF-{kappa}B activation. Recent studies suggest that the evolutionary potential of spike protein is reaching its limit. Indeed, our findings highlight the critical role of non-spike mutations in the future evolution of SARS-CoV-2.
]]></description>
<dc:creator>Tsujino, S.</dc:creator>
<dc:creator>Tsuda, M.</dc:creator>
<dc:creator>Ito, J.</dc:creator>
<dc:creator>Deguchi, S.</dc:creator>
<dc:creator>Taha, T. Y.</dc:creator>
<dc:creator>Nasser, H.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Rosecrans, J.</dc:creator>
<dc:creator>Suzuki, R.</dc:creator>
<dc:creator>Suzuki, S.</dc:creator>
<dc:creator>Yoshimatsu, K.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>The Genotype to Phenotype Japan (G2P-Japan) Consortium,</dc:creator>
<dc:creator>Ikeda, T.</dc:creator>
<dc:creator>Takayama, K.</dc:creator>
<dc:creator>Sato, K.</dc:creator>
<dc:creator>Tanaka, S.</dc:creator>
<dc:creator>Tamura, T.</dc:creator>
<dc:creator>Fukuhara, T.</dc:creator>
<dc:date>2025-05-30</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656516</dc:identifier>
<dc:title><![CDATA[A non-spike nucleocapsid R204P mutation in SARS-CoV-2 Omicron XEC enhances inflammation and pathogenicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.28.656743v1?rss=1">
<title>
<![CDATA[
A Transcriptomics-Based Computational Drug Repositioning Pipeline Identifies Simvastatin And Primaquine As Novel Therapeutics For Endometriosis Pain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.28.656743v1?rss=1</link>
<description><![CDATA[
Endometriosis has limited treatment options, prompting the search for novel therapeutics. We previously used a transcriptomics-based computational drug repositioning pipeline to analyze public bulk transcriptomic data of eutopic endometrium from cases and controls and identified several drug candidates. Fenoprofen, our top in silico candidate, was validated in a rat model of endometriosis-associated pain. Building on this, we evaluated herein two additional candidates, simvastatin (a cholesterol-lowering drug) and primaquine (an antimalarial), based on strong endometrial gene expression reversal scores and favorable safety profiles. Using the rat model, we conducted behavioral testing, bulk RNA sequencing, and differential expression analysis to assess their therapeutic potential. We also assessed endometriosis diagnosis among patients prescribed simvastatin in electronic medical records (EMR) across six University of California (UC) healthcare institutions. In vivo validation using a rat model of endometriosis demonstrated that both simvastatin and primaquine significantly reduced vaginal hyperalgesia, a surrogate marker of endometriosis-related pain. RNA-seq of uteri and lesions confirmed reversal of disease-associated gene expression signatures following treatment. Analysis of UC-wide EMR data found lower relative risk of endometriosis among those prescribed simvastatin compared to a matched control group. Overall, simvastatin and primaquine attenuated pain-associated behaviors and reversed endometriosis-related gene expression changes in an animal model. Moreover, simvastatin prescription was associated with a lower relative risk of endometriosis in our retrospective multi-center cohort study. These findings highlight their potential as repurposed therapeutics for endometriosis and support the effectiveness of computational drug repositioning in identifying new treatment strategies.

One Sentence SummarySimvastatin and primaquine reduced endometriosis pain and reversed gene signatures, with simvastatin also linked to lower disease risk.
]]></description>
<dc:creator>Oskotsky, T. T.</dc:creator>
<dc:creator>Tang, X.</dc:creator>
<dc:creator>Arthurs, E.</dc:creator>
<dc:creator>Govil, A.</dc:creator>
<dc:creator>Abbasi, F.</dc:creator>
<dc:creator>Bhoja, A.</dc:creator>
<dc:creator>Bunis, D. J.</dc:creator>
<dc:creator>Lau, A.</dc:creator>
<dc:creator>Einhaus, J.</dc:creator>
<dc:creator>Diop, M.</dc:creator>
<dc:creator>Irwin, J. C.</dc:creator>
<dc:creator>Gaudilliere, B.</dc:creator>
<dc:creator>Stevenson, D. K.</dc:creator>
<dc:creator>Giudice, L. C.</dc:creator>
<dc:creator>McAllister, S. L.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2025-05-30</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656743</dc:identifier>
<dc:title><![CDATA[A Transcriptomics-Based Computational Drug Repositioning Pipeline Identifies Simvastatin And Primaquine As Novel Therapeutics For Endometriosis Pain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.29.656244v1?rss=1">
<title>
<![CDATA[
Genome-wide CRISPR screens identify PTGES3 as a novel AR modulator 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.29.656244v1?rss=1</link>
<description><![CDATA[
The androgen receptor (AR) is a critical driver of prostate cancer (PCa). To study regulators of AR protein levels and oncogenic activity, we created the first live cell quantitative endogenous AR fluorescent reporters. Leveraging this novel AR reporter, we performed genome-scale CRISPRi flow cytometry sorting screens to systematically identify genes that modulate AR protein levels. We identified and validated known AR protein regulators including HOXB13 and GATA2 and also unexpected top hits including PTGES3, a poorly characterized gene in PCa. PTGES3 repression resulted in loss of AR protein, cell cycle arrest, and cell death in AR-driven PCa models. PTGES3 is not a commonly essential gene, and our data nominate it as a prime PCa therapeutic target. Clinically, analysis of PCa data demonstrate that PTGES3 expression is associated with AR-directed therapy resistance. Mechanistically, we show PTGES3 binds directly to AR, forms a protein complex with AR in the nucleus, regulates AR protein stability in vitro and in vivo and modulates AR function in the nucleus at AR target genes. PTGES3 represents a novel therapeutic target for overcoming known mechanisms of resistance to existing AR-directed therapies in PCa.
]]></description>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Melnyk, J. E.</dc:creator>
<dc:creator>Fu, B. X. H.</dc:creator>
<dc:creator>Shrestha, R.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Sjostrom, M.</dc:creator>
<dc:creator>Feng, S.</dc:creator>
<dc:creator>Anderson, J.</dc:creator>
<dc:creator>Han, W.</dc:creator>
<dc:creator>Chesner, L. N.</dc:creator>
<dc:creator>Shin, H. J.</dc:creator>
<dc:creator>Farsh, T.</dc:creator>
<dc:creator>Suarez, H. J.</dc:creator>
<dc:creator>Nath, S.</dc:creator>
<dc:creator>Chou, J.</dc:creator>
<dc:creator>Das, R.</dc:creator>
<dc:creator>Egusa, E. A.</dc:creator>
<dc:creator>Calvert, M.</dc:creator>
<dc:creator>Kishishita, A.</dc:creator>
<dc:creator>Barpanda, A.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Maheshwari, A.</dc:creator>
<dc:creator>Chen, W. S.</dc:creator>
<dc:creator>Alshalalfa, M.</dc:creator>
<dc:creator>Winters, A.</dc:creator>
<dc:creator>Hua, J. T.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Davicioni, E.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:creator>Stohr, B. A.</dc:creator>
<dc:creator>Siddiqui, J.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Small, E. J.</dc:creator>
<dc:creator>Shokat, K. M.</dc:creator>
<dc:creator>Nelson, P.</dc:creator>
<dc:creator>Quigley, D. A.</dc:creator>
<dc:creator>Wasmuth, E. V.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:creator>Feng, F. Y.</dc:creator>
<dc:date>2025-05-30</dc:date>
<dc:identifier>doi:10.1101/2025.05.29.656244</dc:identifier>
<dc:title><![CDATA[Genome-wide CRISPR screens identify PTGES3 as a novel AR modulator]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.27.656356v1?rss=1">
<title>
<![CDATA[
Complex I Drives Glutamine-Dependent TCA Cycle to Support Viability of MYChigh Breast Cancer Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.27.656356v1?rss=1</link>
<description><![CDATA[
In many cancers, stably elevated MYC levels drive persistent and concerted activation of cell growth promoting anabolic programs and the cell cycle in ways that are distinct from normal cells. Therefore, synthetic-lethal strategies to target MYC reprograming of these pathways may identify new selective anticancer therapies for the treatment of MYChigh tumors. Here, we identify enhanced mitochondrial respiration as a hallmark of MYC overexpressing cancer cells. Mitochondrial respiration sustains the TCA cycle by regenerating NAD+ through complex I-mediated oxidation of NADH. Metabolic carbon tracing analysis revealed that MYC shifts TCA cycles carbon source from glucose to glutamine. Inhibition of the glutamine-fueled TCA cycle using NAD+-depleting complex I inhibitors resulted in MYC-dependent synthetic lethality in breast cancer cells. In mouse models of MYChigh tumors, persistent inhibition of tumor growth was achieved through combined inhibition of complex I and glutaminolysis. Our results suggest that the high respiration rate observed in MYChigh cells supports glutamine carbon-enriched TCA cycle, rendering MYChigh tumors selectively vulnerable to inhibitors of mitochondrial respiration and glutaminolysis.
]]></description>
<dc:creator>Anttila, J. M.</dc:creator>
<dc:creator>Savelius, M.</dc:creator>
<dc:creator>Somani, J.</dc:creator>
<dc:creator>Nicorici, D.</dc:creator>
<dc:creator>Munne, P. M.</dc:creator>
<dc:creator>Id, L.</dc:creator>
<dc:creator>Peura, A.</dc:creator>
<dc:creator>Hiltunen, A. O.</dc:creator>
<dc:creator>Aung, J.</dc:creator>
<dc:creator>Awadhpersad, R.</dc:creator>
<dc:creator>Prajapati, B.</dc:creator>
<dc:creator>Peltonen, M.</dc:creator>
<dc:creator>Ala-Hongisto, H.</dc:creator>
<dc:creator>Gautam, P.</dc:creator>
<dc:creator>Valimaki, M. J.</dc:creator>
<dc:creator>Tervonen, T. A.</dc:creator>
<dc:creator>Sapovalovaite, K.</dc:creator>
<dc:creator>Devarajan, R.</dc:creator>
<dc:creator>Perez, M. V. R.</dc:creator>
<dc:creator>Mutka, M.</dc:creator>
<dc:creator>Kovanen, P.</dc:creator>
<dc:creator>Niinikoski, L.</dc:creator>
<dc:creator>Meretoja, T.</dc:creator>
<dc:creator>Mattson, J.</dc:creator>
<dc:creator>Heikkila, P.</dc:creator>
<dc:creator>Wennerberg, K.</dc:creator>
<dc:creator>Arsenian-Henriksson, M.</dc:creator>
<dc:creator>Westermarck, J.</dc:creator>
<dc:creator>Aittokallio, T.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:creator>Jackson, C. B.</dc:creator>
<dc:creator>Nieminen, A. I.</dc:creator>
<dc:creator>Klefstrom, J.</dc:creator>
<dc:date>2025-05-31</dc:date>
<dc:identifier>doi:10.1101/2025.05.27.656356</dc:identifier>
<dc:title><![CDATA[Complex I Drives Glutamine-Dependent TCA Cycle to Support Viability of MYChigh Breast Cancer Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.30.657075v1?rss=1">
<title>
<![CDATA[
An atlas of TF driven gene programs across human cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.30.657075v1?rss=1</link>
<description><![CDATA[
Combinations of transcription factors (TFs) regulate gene expression and determine cell fate. Much effort has been devoted to understanding TF activity in different tissues and how tissue-specificity is achieved. However, ultimately gene regulation occurs at the single cell level and the recent explosion in the availability of single cell gene expression data now makes it possible to understand TF activity at this granular level of resolution.

Here, we leverage a large collection of Human Cell Atlas (HCA) single cell data to explore TF activity by examining cell-type and tissue-specific sets of target genes, or regulons. We compile a regulon atlas, CellRegulon, and map the activity of TFs in an extensive set of healthy adult and foetal tissues spanning hundreds of cell types. Using CellRegulon, we describe dynamic patterns of co-regulation, associate TF-modules with different cellular functions and characterise the distribution of active TFs and TF families across cell types. We show that CellRegulon can link disease gene expression signatures to cell types and TFs relevant to the disease. Finally, using a newly generated multiome dataset of the adult lung, we show how CellRegulon can be extended into an enhancer-gene regulatory network (eGRN) to improve cell-type associations with genetic risk loci for diseases, such as childhood onset asthma, COPD and IPF, and to identify high risk gene modules. Our database for easy download and interactive exploration allows researchers to understand key gene modules activated at cell type transitions and will therefore be valuable for tasks such as cell type engineering (https://www.cellregulondb.org).
]]></description>
<dc:creator>Pett, J. P.</dc:creator>
<dc:creator>Prete, M.</dc:creator>
<dc:creator>Pham, D.</dc:creator>
<dc:creator>England, N.</dc:creator>
<dc:creator>Yuan, H.</dc:creator>
<dc:creator>Prigmore, E.</dc:creator>
<dc:creator>Tuck, L.</dc:creator>
<dc:creator>Oszlanczi, A.</dc:creator>
<dc:creator>To, K.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Suo, C.</dc:creator>
<dc:creator>Dann, E.</dc:creator>
<dc:creator>He, P.</dc:creator>
<dc:creator>Kedlian, V. R.</dc:creator>
<dc:creator>Kanemaru, K.</dc:creator>
<dc:creator>Cranley, J.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Elmentaite, R.</dc:creator>
<dc:creator>Oliver, A. J.</dc:creator>
<dc:creator>Cujba, A.-M.</dc:creator>
<dc:creator>Cakir, B.</dc:creator>
<dc:creator>Murray, S.</dc:creator>
<dc:creator>Mahbubani, K. T.</dc:creator>
<dc:creator>Saeb-Parsy, K.</dc:creator>
<dc:creator>Gambardella, L.</dc:creator>
<dc:creator>Kasper, M.</dc:creator>
<dc:creator>Haniffa, M. A.</dc:creator>
<dc:creator>Nawijn, M. C.</dc:creator>
<dc:creator>Teichmann, S. A.</dc:creator>
<dc:creator>Meyer, K. B.</dc:creator>
<dc:date>2025-06-01</dc:date>
<dc:identifier>doi:10.1101/2025.05.30.657075</dc:identifier>
<dc:title><![CDATA[An atlas of TF driven gene programs across human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.01.657285v1?rss=1">
<title>
<![CDATA[
Active learnings impact on student course performance in STEM varies by type and intensity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.01.657285v1?rss=1</link>
<description><![CDATA[
We updated a recent meta-analysis of active learnings impact on student achievement in undergraduate STEM courses by following the same protocol to evaluate studies published from 2010-2017. We screened 1659 papers, coded 1294, and found 210 that met five pre-established inclusion criteria and six pre-established criteria for methodological quality. After further dropping 76 studies with no exam scores data, 134 of these studies contained data on student performance on identical or equivalent exams. We found that on average, active learnings effect size on exam scores was 0.519 {+/-} 0.049, meaning that when students are in active learning classes, they perform roughly half a standard deviation higher on an identical exam. Funnel plots and sensitivity analyses indicated that these results were not due to sampling bias. Active learning had a positive impact on student outcomes regardless of class size, course level, or STEM discipline, though there was heterogeneity in the effects. All of these results are very similar when compared to earlier meta-analyses, however increased resolution in the studies analyzed here revealed two novel results. First, student performance was significantly better in courses that employed high-intensity active learning, defined as students being on task at least two-thirds of class time, versus lower-intensities. Additionally, there was significant heterogeneity in efficacy across different types of active learning employed. These results suggest that most, if not all types of active learning are effective, and that when innovating in their classes, instructors should continually work to increase active learning intensity. We urge caution in interpreting the results on active learning types, however, and propose a preliminary framework for making more-sophisticated and reliable analyses of variation in course design. Finally, the evidence presented here for active learnings impact on student outcomes creates a strong foundation for faculty professional development and administration.
]]></description>
<dc:creator>Xu, S.</dc:creator>
<dc:creator>Velasco, V.</dc:creator>
<dc:creator>Hill, M. J.</dc:creator>
<dc:creator>Tran, E.</dc:creator>
<dc:creator>Agrawal, S.</dc:creator>
<dc:creator>Arroyo, E. N.</dc:creator>
<dc:creator>Behling, S.</dc:creator>
<dc:creator>Chambwe, N.</dc:creator>
<dc:creator>Cintron, D. L.</dc:creator>
<dc:creator>Cooper, J. D.</dc:creator>
<dc:creator>Dunster, G.</dc:creator>
<dc:creator>Grummer, J. A.</dc:creator>
<dc:creator>Hennessey, K.</dc:creator>
<dc:creator>Hsiao, J.</dc:creator>
<dc:creator>Iranon, N.</dc:creator>
<dc:creator>Jones, L.</dc:creator>
<dc:creator>Jordt, H.</dc:creator>
<dc:creator>Keller, M.</dc:creator>
<dc:creator>Lacey, M. E.</dc:creator>
<dc:creator>Littlefield, C. E.</dc:creator>
<dc:creator>Lowe, A.</dc:creator>
<dc:creator>Newman, S.</dc:creator>
<dc:creator>Okolo, V.</dc:creator>
<dc:creator>Olroyd, S.</dc:creator>
<dc:creator>Peecook, B. R.</dc:creator>
<dc:creator>Pickett, S. B.</dc:creator>
<dc:creator>Slager, D. L.</dc:creator>
<dc:creator>Caviedes-Solis, I. W.</dc:creator>
<dc:creator>Stanchak, K. E.</dc:creator>
<dc:creator>Sundaravaradan, V.</dc:creator>
<dc:creator>Valdebenito, C.</dc:creator>
<dc:creator>Williams, C. R.</dc:creator>
<dc:creator>Zinsli, K. A.</dc:creator>
<dc:creator>Freeman, S.</dc:creator>
<dc:creator>Theobald, E. J.</dc:creator>
<dc:date>2025-06-02</dc:date>
<dc:identifier>doi:10.1101/2025.06.01.657285</dc:identifier>
<dc:title><![CDATA[Active learnings impact on student course performance in STEM varies by type and intensity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.31.657194v1?rss=1">
<title>
<![CDATA[
Clonal Expansion of Alveolar Fibroblast Progeny Drives Pulmonary Fibrosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.31.657194v1?rss=1</link>
<description><![CDATA[
Pulmonary fibrosis has been called a fibroproliferative disease but the functional importance of proliferating fibroblasts to pulmonary fibrosis has not been systematically examined. In response to alveolar injury, resting alveolar fibroblasts differentiate into fibrotic fibroblasts that express high levels of collagens. However, what role, if any, proliferation plays in the accumulation of fibrotic fibroblasts remains unclear. Through EdU incorporation, genetic lineage tracing, and single cell RNA sequencing, we resolve the proliferation dynamics of lung fibroblasts during post-injury fibrogenesis. Our data show substantial DNA replication in progeny of alveolar fibroblasts in two models of pulmonary fibrosis. By genetically labeling individual cells, we observe clonal expansion of alveolar fibroblast descendants principally in regions of fibrotic remodeling. The transcriptome of proliferating fibroblasts closely resembles that of fibrotic fibroblasts, suggesting that fibroblasts can first differentiate into fibrotic fibroblasts and then proliferate. Genetic ablation of proliferating fibroblasts and selective inhibition of cytokinesis in alveolar fibroblast descendants significantly mitigates pulmonary fibrosis and rescues lung function. Furthermore, fibroblasts in precision-cut lung slices from human fibrotic lungs exhibit higher proliferation rates than those in non-diseased lungs. This work establishes fibroblast proliferation as a critical driver of pulmonary fibrosis and suggests that specifically targeting fibroblast proliferation could be a new therapeutic strategy for fibrotic diseases.
]]></description>
<dc:creator>Molina, C.</dc:creator>
<dc:creator>Tsukui, T.</dc:creator>
<dc:creator>Khan, I.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Qiu, W.</dc:creator>
<dc:creator>Matthay, M.</dc:creator>
<dc:creator>Wolters, P.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:date>2025-06-03</dc:date>
<dc:identifier>doi:10.1101/2025.05.31.657194</dc:identifier>
<dc:title><![CDATA[Clonal Expansion of Alveolar Fibroblast Progeny Drives Pulmonary Fibrosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.04.657671v1?rss=1">
<title>
<![CDATA[
Clinical Trypanosoma cruzi isolates share a common antigen repertoire that is absent from culture adapted strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.04.657671v1?rss=1</link>
<description><![CDATA[
BackgroundTrypanosoma cruzi causes Chagas disease, a poorly understood and clinically heterogeneous disease. Recent work has demonstrated that parasites adapted to laboratory conditions are genomically variable, but little is known of the extent of genomic diversity from clinically isolated specimens.

MethodsIn this retrospective observational genomic study, we isolated 15 T. cruzi specimens from three clinical studies of Chagas disease, representing different clinical contexts. We sequenced the genome of each strain and used single nucleotide variant (SNV) based analyses to estimate parasite genetic lineage, genomic population structure, regions of copy number plasticity, and to identify gene conversion events. In addition, we generated and annotated whole genome assemblies of each isolate. From these assemblies, we compared the repertoires of genes encoding for highly virulent and variable proteins that have been implicated in disease pathogenesis.

FindingsWe identified parasites from two genetic lineages in this collection of clinical isolates. Our analysis revealed evidence of genomic instability. Diversity-generating copy number variation was statistically enriched in regions encoding the virulence-associated multigene families, while diversity-eliminating gene conversion events were enriched in regions depleted of multigene family members. We also discovered a set of multigene family members that is present in all of the clinically isolated parasite genomes and absent from all of the lab adapted strains, regardless of parasite lineage. Multigene family repertoires were more conserved among field isolated specimens of the same genetic lineage than among culture adapted strains of the same genetic type.

InterpretationThis study provides whole genome sequencing data for TcV parasites isolated from naturally infected human patients with Chagas disease for the first time. Our analysis of these genomes revealed substantial genomic instability, suggesting the parasite undergoes genomic change in response to the pressures imposed by the host environment. Moreover, we observed a set of virulence-associated genes that are present exclusively within clinical isolates and absent from lab-adapted strains, indicating a potential role for these genes in parasite survival in natural hosts. These findings highlight the limitations of genetic studies focused exclusively on lab-adapted parasite strains and provide insight into the genomic features of T. cruzi that are likely to be important for clinical infection.
]]></description>
<dc:creator>Hakim, J. M. C.</dc:creator>
<dc:creator>Gutierrez Guarnizo, S. A.</dc:creator>
<dc:creator>Duran, A.</dc:creator>
<dc:creator>Malaga-Machacha, E.</dc:creator>
<dc:creator>Duque, C.</dc:creator>
<dc:creator>Singer, L.</dc:creator>
<dc:creator>Colanzi, R.</dc:creator>
<dc:creator>Sherbuk, J. E.</dc:creator>
<dc:creator>Gilman, R.</dc:creator>
<dc:creator>Bern, C.</dc:creator>
<dc:creator>Messenger, L. A.</dc:creator>
<dc:creator>Mugnier, M. R.</dc:creator>
<dc:creator>Working Group on Chagas Disease in Bolivia and Peru,</dc:creator>
<dc:date>2025-06-04</dc:date>
<dc:identifier>doi:10.1101/2025.06.04.657671</dc:identifier>
<dc:title><![CDATA[Clinical Trypanosoma cruzi isolates share a common antigen repertoire that is absent from culture adapted strains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.02.657515v1?rss=1">
<title>
<![CDATA[
An improved model for prediction of de novo designed proteins with diverse geometries 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.02.657515v1?rss=1</link>
<description><![CDATA[
Nature uses structural variations on protein folds to fine-tune the geometries of proteins for diverse functions, yet deep learning-based de novo protein design methods generate highly regular, idealized protein fold geometries that fail to capture natural diversity. Here, using physics-based design methods, we generated and experimentally validated a dataset of 5,996 stable, de novo designed proteins with diverse non-ideal geometries. We show that deep learning-based structure prediction methods applied to this set have a systematic bias towards idealized geometries. To address this problem, we present a fine-tuned version of Alphafold2 that is capable of recapitulating geometric diversity and generalizes to a new dataset of thousands of geometrically diverse de novo proteins from 5 fold families unseen in fine-tuning. Our results suggest that current deep learning-based structure prediction methods do not capture some of the physics that underlie the specific conformational preferences of proteins designed de novo and observed in nature. Ultimately, approaches such as ours and further informative datasets should lead to improved models that reflect more of the physical principles of atomic packing and hydrogen bonding interactions and enable improved generalization to more challenging design problems.
]]></description>
<dc:creator>Orr, B.</dc:creator>
<dc:creator>Crilly, S. E.</dc:creator>
<dc:creator>Akpinaroglu, D.</dc:creator>
<dc:creator>Zhu, E.</dc:creator>
<dc:creator>Keiser, M. J.</dc:creator>
<dc:creator>Kortemme, T.</dc:creator>
<dc:date>2025-06-06</dc:date>
<dc:identifier>doi:10.1101/2025.06.02.657515</dc:identifier>
<dc:title><![CDATA[An improved model for prediction of de novo designed proteins with diverse geometries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.03.657754v1?rss=1">
<title>
<![CDATA[
IL-18 inhibition enlarges lesions, necrotic cores and thickens fibrous caps in Jak2V617F clonal hematopoiesis-driven atherosclerosis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.03.657754v1?rss=1</link>
<description><![CDATA[
BackgroundInflammasome activation promotes atherosclerosis in clonal hematopoiesis (CH). Active inflammasomes secrete both IL-1{beta} and IL-18. Plasma IL-18 levels are elevated in Jak2VFCH. Genetic deficiency of IL-18 has been shown to reduce atherosclerosis in non-CH murine models. However, whether IL-18 inhibition promotes atherosclerosis in control or Jak2VF CH is unknown.

Methods and resultsLdlr-/- mice were transplanted with bone marrow (BM) from Mx1-cre Jak2VF (20%) and wild-type (80%) mice or with control BM, fed a Western-type diet (WTD) for 8, 10 or 16 weeks and administered control or IL-18 IgG from 4 weeks onwards.

IL-18 antibody treatment increased plaque collagen content and cap thickness. Unexpectedly, IL-18 antibody treatment increased the size of early lesions and promoted formation of advanced lesions with large necrotic cores in Jak2VF CH mice. IL-18 antibody treatment was associated with diminished interferon (IFN)-{gamma} and AIM2 levels and reduced macrophage pyroptosis especially in Jak2VF CH mice. However, IL-18 antibodies increased cleaved Caspase-3 and TUNEL+ macrophages (indicating increased apoptosis) and reduced efferocytosis. Sc-RNA-seq analysis showed that IL-18 antibody treatment reduced expression of MHC class II genes, a marker of IFN-{gamma} signaling, and of genes mediating efferocytosis (Mertk and Axl), in resident-like macrophage subpopulations in Jak2VF CH mice. Consistently, IFN-{gamma} injection increased Axl and Mertk expression in resident peritoneal macrophages.

ConclusionsDespite improvements in collagen and fibrous cap thickness in Jak2VF CH mice, IL-18 antibody treatment increased advanced necrotic lesions, reflecting a shift from pyroptotic to apoptotic cell death coupled with defective efferocytosis, events which were coordinated by reduced IFN-{gamma} signaling. These findings indicate a mixed atherosclerosis phenotype resulting from IL-18 inhibition, advocating for alternative therapeutic strategies.

Inhibition of IL-18 has been considered as a novel therapeutic approach to reduce atherosclerosis and stabilize atherosclerotic plaques. We show that IL-18 antibodies have adverse effects on atherosclerotic lesional necrosis, calling this approach into question.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=169 SRC="FIGDIR/small/657754v1_ufig1.gif" ALT="Figure 1">
View larger version (33K):
org.highwire.dtl.DTLVardef@a93b2forg.highwire.dtl.DTLVardef@6cc057org.highwire.dtl.DTLVardef@1c7afc5org.highwire.dtl.DTLVardef@e4da82_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LIInflammasome activation produces active IL-1 and IL-18 and worsens atherosclerosis in clonal hematopoiesis (CH) however the contribution of IL-18 is unknown.
C_LIO_LIAntibody inhibition of IL-18 increased plaque collagen but also increased early lesion area and late lesions with large necrotic cores in Jak2VF CH mice.
C_LIO_LIThere was a reversal of AIM2 inflammasome activation but a switch to apoptosis which along with reduced efferocytosis increased necrosis
C_LIO_LIThese events appeared to be coordinated by reduced IFN-{gamma} which increased collagen but also decreased expression of efferocytotic genes. Our studies call into question whether inhibition of IL-18 would stabilize plaques in CH.
C_LI
]]></description>
<dc:creator>Tavallaie, M.</dc:creator>
<dc:creator>Hsu, C.-C.</dc:creator>
<dc:creator>Hardaway, B.</dc:creator>
<dc:creator>Dou, H.</dc:creator>
<dc:creator>Fidler, T.</dc:creator>
<dc:creator>Kim, E.</dc:creator>
<dc:creator>Wang, N.</dc:creator>
<dc:creator>Westerterp, M.</dc:creator>
<dc:creator>Tall, A. R.</dc:creator>
<dc:date>2025-06-06</dc:date>
<dc:identifier>doi:10.1101/2025.06.03.657754</dc:identifier>
<dc:title><![CDATA[IL-18 inhibition enlarges lesions, necrotic cores and thickens fibrous caps in Jak2V617F clonal hematopoiesis-driven atherosclerosis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.04.657796v1?rss=1">
<title>
<![CDATA[
AlphaFold models of host-pathogen interactions elucidate the prevalence and structural modes of molecular mimicry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.04.657796v1?rss=1</link>
<description><![CDATA[
Pathogens exploit host cellular machinery through protein-protein interactions (PPIs), often using molecular mimicry to hijack host cellular processes. While there have been thousands of host-pathogen PPIs determined to date, the lack of structural information for these impedes the study of the prevalence of molecular mimicry and convergent evolution of protein interaction interfaces. To address this, we benchmarked AlphaFold2 and 3 for prediction of structures of host-pathogen interactions observing that accurate models can be retrieved when ranking by modelling confidence, despite an overall low performance. We predicted structures for 6,782 pathogen-human PPIs yielding 803 models of higher confidence. Most pathogen proteins interacting with a common human protein are predicted to do so via the same interface, suggesting a high degree of convergent evolution of protein interaction interfaces. When comparing structural models from host-pathogen and host-host interactions, we observe that a majority of pathogen proteins are predicted to target existing human PPI interfaces. We categorized instances of mimicry into different modes, occurring at different frequencies: 1) via the same domain family (least common); 2) via a similar structural motif; and 3) via a similar linear motif (most common). We selected examples of linear motif interactions for binding assay testing, confirming 8 out of 12 predicted interfaces, including 3 viral linear motif interactions. This validates AlphaFolds ability to model some host-pathogen interactions and the mechanisms underlying molecular mimicry. This work showcases the value of large-scale structural modelling to study convergent evolution of host-pathogen interactions and how molecular mimicry may contribute to infection or host defense.
]]></description>
<dc:creator>Baptista, D.</dc:creator>
<dc:creator>Gomez-Lucas, L.</dc:creator>
<dc:creator>Jänes, J.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Joao Amorim, M.</dc:creator>
<dc:creator>Ivarsson, Y.</dc:creator>
<dc:creator>Beltrao, P.</dc:creator>
<dc:date>2025-06-06</dc:date>
<dc:identifier>doi:10.1101/2025.06.04.657796</dc:identifier>
<dc:title><![CDATA[AlphaFold models of host-pathogen interactions elucidate the prevalence and structural modes of molecular mimicry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.05.657915v1?rss=1">
<title>
<![CDATA[
Mechanical force locally damages, remodels and stabilizes the lattice of spindle microtubules 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.05.657915v1?rss=1</link>
<description><![CDATA[
To segregate chromosomes at cell division, the spindle must maintain its structure under force. How it does so remains poorly understood. To address this question, we use microneedle manipulation to apply local force to spindle microtubule bundles, kinetochore-fibers (k-fibers), inside mammalian cells. We show that local load directly fractures k-fibers, and that newly created plus-ends often have arrested dynamics, resisting depolymerization. Force alone, without fracture, is sufficient for spindle microtubule stabilization, as revealed by laser ablating k-fibers under local needle force. Doublecortin, which binds a compacted microtubule lattice, is lost around the force application site, suggesting local force-induced structural remodeling. In turn, EB1, which recognizes GTP-tubulin, is locally enriched at stabilization sites, both before and after force-induced fracture. Together, our findings support a model where force-induced damage leads to local spindle microtubule lattice remodeling and stabilization, which we propose reinforces the spindle where it experiences critical loads.
]]></description>
<dc:creator>Rux, C. J.</dc:creator>
<dc:creator>Chong, M. K.</dc:creator>
<dc:creator>Myers, S.</dc:creator>
<dc:creator>Cho, N. H.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2025-06-06</dc:date>
<dc:identifier>doi:10.1101/2025.06.05.657915</dc:identifier>
<dc:title><![CDATA[Mechanical force locally damages, remodels and stabilizes the lattice of spindle microtubules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.05.658176v1?rss=1">
<title>
<![CDATA[
Regulators of Interferon-Responsive Microglia Uncovered by Genome-wide CRISPRi Screening 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.05.658176v1?rss=1</link>
<description><![CDATA[
Microglia dynamically support brain homeostasis through the induction of specialized activation programs or states. One such program is the Interferon-Responsive Microglia state (IRM), which has been identified in developmental windows, aging, and disease. While the functional importance of this state is becoming increasingly clear, our understanding of the regulatory networks that govern IRM induction remain incomplete. To systematically identify genetic regulators of the IRM state, we conducted a genome-wide CRISPR interference (CRISPRi) screen in human iPSC-derived microglia (iPS-Microglia) using IFIT1 as a representative IRM marker. We identified 772 genes that modulate IRM, including canonical type I interferon signaling genes (IFNAR2, TYK2, STAT1/2, USP18) and novel regulators. We uncovered a non-canonical role for the CCR4-NOT complex subunit CNOT10 in IRM activation, independent of its traditional function. This work provides a comprehensive resource for dissecting IRM biology and highlights both established and novel targets for modulating microglial interferon signaling in health and disease.
]]></description>
<dc:creator>McQuade, A.</dc:creator>
<dc:creator>Castillo, V. C.</dc:creator>
<dc:creator>Hagan, V.</dc:creator>
<dc:creator>Liang, W.</dc:creator>
<dc:creator>Ta, T.</dc:creator>
<dc:creator>Mishra, R.</dc:creator>
<dc:creator>Teter, O.</dc:creator>
<dc:creator>Teyssier, N.</dc:creator>
<dc:creator>Leng, K.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:date>2025-06-06</dc:date>
<dc:identifier>doi:10.1101/2025.06.05.658176</dc:identifier>
<dc:title><![CDATA[Regulators of Interferon-Responsive Microglia Uncovered by Genome-wide CRISPRi Screening]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.04.657687v1?rss=1">
<title>
<![CDATA[
Live-cell analyses with unsegmented images to study cancer cell response to modified T cell therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.04.657687v1?rss=1</link>
<description><![CDATA[
Live-cell imaging (LCI) of modified T cells co-cultured with cancer cells is commonly used to quantify T cell anti-cancer function. Videos captured by LCI show complex multi-cell behavioral phenotypes that go beyond simple cancer cell fluorescence measurements. Here, we develop an unsupervised analysis workflow to characterize LCI data generated using the Incucyte imaging platform. Unlike most LCI analyses, we avoid cell segmentation due to the low spatiotemporal resolution of the LCI videos and high levels of cell-cell contact. Instead, we develop methods that identify global aggregation patterns and local cellular keypoints to characterize the multicellular interactions that determine cancer cell sensitivity to, or escape from, T cell surveillance. We demonstrate our segmentation-free live-cell behavioral analysis (SF-LCBA) methods on TCR T cells from four donors with varying proportions of cells with a beneficial RASA2 knockout and effector-to-target initial concentrations in a co-culture with A375 melanoma cells. We find that different T cell modifications affect the spatiotemporal dynamics of multicellular aggregate formation. In particular, we show that fewer and smaller cancer cell aggregates form at high ratios of effector T cells to target cancer cells and high titrations of T cells with RASA2 knockouts. Our SF-LCBA method identifies, characterizes, and tracks cellular aggregate formation in datasets that are unsuitable for cell segmentation and tracking, opening the door to more therapeutically-relevant measurements of modified T cell therapy cell behavioral phenotypes from LCI data.
]]></description>
<dc:creator>Epstein, L.</dc:creator>
<dc:creator>Weiner, A. C.</dc:creator>
<dc:creator>Verma, A.</dc:creator>
<dc:creator>Saedi, M.</dc:creator>
<dc:creator>Carnevale, J.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Engelhardt, B. E.</dc:creator>
<dc:date>2025-06-07</dc:date>
<dc:identifier>doi:10.1101/2025.06.04.657687</dc:identifier>
<dc:title><![CDATA[Live-cell analyses with unsegmented images to study cancer cell response to modified T cell therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.06.658239v1?rss=1">
<title>
<![CDATA[
Paracrine Signals from HIV-1 Infected Immune Cells Reprogram Cervical Cancer Pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.06.658239v1?rss=1</link>
<description><![CDATA[
Persistent infection with human papillomavirus (HPV) is the primary cause of cervical cancer worldwide. Notably, women co-infected with HPV and human immunodeficiency virus type 1 (HIV-1) have a six-fold higher lifetime risk of developing cervical cancer compared to those without HIV, even when adhering to antiretroviral therapy (ART) and achieving T-cell reconstitution. While chronic HIV-1 infection is known to cause inflammation, how paracrine signals from immune cells alter signaling in cervical cells remain poorly understood. To address this, we conducted global transcriptomics analysis on cervical swabs from Kenyan women with HPV, stratified by HIV-1 and cancer status. Strikingly, women with HIV-1 showed cancer-like gene expression patterns in non-cancerous cervical epithelial cells. Complementary global mass spectrometry (MS) proteomics of cervical cells exposed to the secretome of HIV-1 infected primary CD4+ T-cells revealed altered expression of proteins in MAPK, PI3K-AKT, and {beta}-catenin signaling pathways. Integrative network analyses of transcriptomic and proteomic datasets revealed that HIV-1 altered gene expression in key pathways known to drive cervical cancer, including genes commonly mutated in HIV-1-naive disease. Notably, IRS-1, a key PI3K-AKT pathway activator, was found to be consistently upregulated in both participant samples and cell culture models, as were interferon-stimulated genes. Phosphoproteomics MS analysis confirmed PI3K-AKT pathway activation in cervical cells exposed to conditioned media from HIV-1-infected T-cells. Together, our findings uncover how HIV-1 reshapes cervical cell signaling via paracrine mechanisms and highlights the PI3K pathway as a potential therapeutic target in HIV-associated cervical cancer.
]]></description>
<dc:creator>Olwal, C. O.</dc:creator>
<dc:creator>Rathore, U.</dc:creator>
<dc:creator>Makanani, S.</dc:creator>
<dc:creator>Kaushal, P.</dc:creator>
<dc:creator>Ashley, I. A.</dc:creator>
<dc:creator>Ummadi, M. R.</dc:creator>
<dc:creator>Appiah, V.</dc:creator>
<dc:creator>Djomkam Zune, A. L.</dc:creator>
<dc:creator>Blanc, S.</dc:creator>
<dc:creator>Winters, D.</dc:creator>
<dc:creator>Delgado, Y.</dc:creator>
<dc:creator>Muthoka, K.</dc:creator>
<dc:creator>Fabius, J. M.</dc:creator>
<dc:creator>Eckhardt, M.</dc:creator>
<dc:creator>Kaake, R. M.</dc:creator>
<dc:creator>Su, M.</dc:creator>
<dc:creator>Fregoso, O. I.</dc:creator>
<dc:creator>Hultquist, J. F.</dc:creator>
<dc:creator>Orango, E. O.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Kyei, G. B.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Quashie, P. K.</dc:creator>
<dc:creator>Bediako, Y.</dc:creator>
<dc:creator>Bouhaddou, M.</dc:creator>
<dc:date>2025-06-07</dc:date>
<dc:identifier>doi:10.1101/2025.06.06.658239</dc:identifier>
<dc:title><![CDATA[Paracrine Signals from HIV-1 Infected Immune Cells Reprogram Cervical Cancer Pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.04.657810v1?rss=1">
<title>
<![CDATA[
Effects of diffusion MRI spatial resolution on human brain short-range association fiber reconstruction and structural connectivity estimation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.04.657810v1?rss=1</link>
<description><![CDATA[
Short-range association fibers (SAFs) are critical for cortical communications but are often underestimated in conventional resolution diffusion magnetic resonance imaging (dMRI) since they locate within a [~]1.5mm thin layer of superficial white matter. With the emergence of high-resolution diffusion imaging techniques, this study timely evaluated the effects of image spatial resolution on SAF reconstruction using simulation data and multi-resolution (2, 1.5, and 0.96 mm iso.) empirical data acquired on the same 20 healthy subjects using the gSlider sequence and 20 widely used tractography approaches. Resolution effects were qualitatively assessed through model fitting and tractography results and quantitatively evaluated using global and regional short-range connectivity strength (SCS). It is found that lower resolution systematically reduces SCS across all methods in a spatially varying manner. Moreover, tractography methods exhibit significant differences in resolution sensitivity, with diffusion tensor imaging (DTI) based single-tissue single-fiber tractography showing greater vulnerability than constrained spherical deconvolution (CSD)-based multi-tissue multi-fiber tractography. Probabilistic tracking with anatomical constraints (ACT) and filtering (SIFT) improves robustness. Finally, up-sampling data to nominally higher resolution partially mitigates resolution-induced degradation and improves SAF reconstruction accuracy, particularly for DTI tractography. Based on these findings, higher resolution and multi-shell imaging is recommended if possible. For a given dataset, data up-sampling and DTI-based probabilistic tracking with ACT is recommended for single-shell low b-value data. CSD-based probabilistic tracking with ACT and SIFT is recommended for single-shell higher b-value data and multi-shell data. In summary, this study systematically and quantitatively evaluated resolution effects on SAF reconstruction and structural connectivity estimation and provided practical guidelines for more accurate mapping of SAFs. These advances hold promises to improve the characterization of healthy and diseased human brains in a wide range of neuroscientific and clinical applications.
]]></description>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Zhong, W.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Liao, C.</dc:creator>
<dc:creator>David, S. H.</dc:creator>
<dc:creator>Huang, S. Y.</dc:creator>
<dc:creator>Tian, Q.</dc:creator>
<dc:date>2025-06-09</dc:date>
<dc:identifier>doi:10.1101/2025.06.04.657810</dc:identifier>
<dc:title><![CDATA[Effects of diffusion MRI spatial resolution on human brain short-range association fiber reconstruction and structural connectivity estimation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.06.658366v1?rss=1">
<title>
<![CDATA[
Low-density lipoprotein receptor-targeting chimeras for membrane protein degradation and enhanced drug delivery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.06.658366v1?rss=1</link>
<description><![CDATA[
Antibody-based therapeutics encompass diverse modalities for targeting tumor cells. Among these, antibody-drug conjugates (ADCs) and extracellular targeted protein degradation (eTPD) specifically depend on efficient lysosomal trafficking for activity. However, many tumor antigens exhibit poor internalization, limiting ADC effectiveness. To address this, we developed low-density lipoprotein receptor-targeting chimeras (LIPTACs), leveraging the constitutive endocytic and recycling activity of the LDLR to enhance lysosomal delivery. LIPTACs enable efficient and selective degradation of diverse extracellular membrane proteins. Additionally, by coupling LIPTACs with cytotoxic payloads to generate degrader-drug conjugates, we can achieve superior intracellular delivery and enhanced cytotoxicity compared to conventional ADCs. The dual modality addresses key challenges of inadequate internalization in conventional ADCs and cytotoxic potency for current eTPD strategies. Our findings demonstrate that LDLR-mediated trafficking can enhance eTPD and ADCs, providing a hybrid blueprint for developing next-generation antibody therapeutics with broader utility and improved efficacy in cancer treatment.
]]></description>
<dc:creator>Zhao, F.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Schaefer, K.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Miao, K.</dc:creator>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Ganjave, S. D.</dc:creator>
<dc:creator>Kumru, K.</dc:creator>
<dc:creator>Peters-Clarke, T. M.</dc:creator>
<dc:creator>Inague, A.</dc:creator>
<dc:creator>Olzmann, J. A.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2025-06-09</dc:date>
<dc:identifier>doi:10.1101/2025.06.06.658366</dc:identifier>
<dc:title><![CDATA[Low-density lipoprotein receptor-targeting chimeras for membrane protein degradation and enhanced drug delivery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.09.657642v1?rss=1">
<title>
<![CDATA[
Alterations of the composition and spatial organization of the microenvironment following non-dysplastic Barrett's esophagus through progression to cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.09.657642v1?rss=1</link>
<description><![CDATA[
Barretts esophagus (BE), a metaplastic condition that is the only known precursor for esophageal adenocarcinoma (EAC), is relatively common, but progression to cancer is infrequent. BE is inflamed but the contribution of the immune system to the carcinogenic process is unknown. To this end, we contrasted non-dysplastic metaplasia of BE patients, captured when they did not progress (non-progressors), did subsequently, but had not yet progressed (pre-progressors) or had already progressed to EAC (progressors). Using spatial multiplexed 56-protein analysis, serial laser capture microdissection (LCM) RNAseq and shallow whole genome sequencing, we identified prooncogenic immune neighbourhoods and dysregulated immune cell populations predictive of subsequent progression to EAC. Indeed, spatial analysis revealed that M1 macrophages, regulatory natural killer (NK) cells, neutrophils and altered ratios of intraepithelial CD4+ and CD8+ lymphocytes typify tumor microenvironmental (TME) changes associated with cancer initiation. Spatially derived cell-to-cell interactions revealed progression-specific immune cell interaction signatures predominantly involving M1 macrophages NK cells and plasma cells. Furthermore, LCM RNAseq analysis identified gene expression  hot signatures enriched in pre-progression and progression samples. Notably, we also observed a correlation between immune cells and copy number alterations in progressor metaplasia. By exposing coordinated changes in the immune cell landscape in patients at high risk of developing EAC, this multi-omic dataset provides novel diagnostic and therapeutic opportunities
]]></description>
<dc:creator>Lin, M.-L.</dc:creator>
<dc:creator>Hickey, J. W.</dc:creator>
<dc:creator>Passman, A.</dc:creator>
<dc:creator>Carlotti, E.</dc:creator>
<dc:creator>Devkumar, S.</dc:creator>
<dc:creator>Derwa, Y. H.</dc:creator>
<dc:creator>ChinAleong, J.</dc:creator>
<dc:creator>Hackett, R. J.</dc:creator>
<dc:creator>Evans, J. A.</dc:creator>
<dc:creator>Sangwan, V.</dc:creator>
<dc:creator>Novelli, M. R.</dc:creator>
<dc:creator>Rodriguez-Justo, M.</dc:creator>
<dc:creator>Wright, N. A.</dc:creator>
<dc:creator>Jansen, M.</dc:creator>
<dc:creator>Underwood, T. J.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Gascard, P.</dc:creator>
<dc:creator>McLean, M. H.</dc:creator>
<dc:creator>Schurch, C. M.</dc:creator>
<dc:creator>Tlsty, T. M.</dc:creator>
<dc:creator>Ferri, L.</dc:creator>
<dc:creator>Nolan, G. P.</dc:creator>
<dc:creator>McDonald, S. A.</dc:creator>
<dc:date>2025-06-12</dc:date>
<dc:identifier>doi:10.1101/2025.06.09.657642</dc:identifier>
<dc:title><![CDATA[Alterations of the composition and spatial organization of the microenvironment following non-dysplastic Barrett's esophagus through progression to cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.10.658786v1?rss=1">
<title>
<![CDATA[
Multi-selective RAS(ON) Inhibition Targets Oncogenic RAS Mutations and Overcomes RAS/MAPK-Mediated Resistance to FLT3 and BCL2 Inhibitors in Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.10.658786v1?rss=1</link>
<description><![CDATA[
Aberrant activation of the RAS/MAPK signaling limits the clinical efficacy of several targeted therapies in acute myeloid leukemia (AML). In FLT3-mutant AML, the selection of clones harboring heterogeneous RAS mutations drives resistance to FLT3 inhibitors (FLT3i). RAS activation is also associated with resistance to other AML targeted therapies, including the BCL2 inhibitor venetoclax. Despite the critical need to inhibit RAS/MAPK signaling in AML, no targeted therapies have demonstrated clinical benefit in RAS-driven AML. To address this unmet need, we investigated the preclinical activity of RMC-7977, a multi-selective inhibitor of GTP-bound active [RAS(ON)] isoforms of mutant and wild-type RAS in AML models. RMC-7977 exhibited potent antiproliferative and pro-apoptotic activity across AML cell lines with MAPK-activating signaling mutations. In cell line models with acquired FLT3i resistance due to secondary RAS mutations, treatment with RMC-7977 restored sensitivity to FLT3i. Similarly, RMC-7977 effectively reversed resistance to venetoclax in RAS-addicted cell line models with both RAS wild-type and mutant genetic backgrounds. In murine patient-derived xenograft models of RAS-mutant AML, RMC-7977 was well tolerated and significantly suppressed leukemic burden in combination with gilteritinib or venetoclax. Our findings strongly support clinical investigation of broad-spectrum RAS(ON) inhibition in AML to treat and potentially prevent drug resistance due to activated RAS signaling.
]]></description>
<dc:creator>Popescu, B.</dc:creator>
<dc:creator>Jones, M. F.</dc:creator>
<dc:creator>Piao, M.</dc:creator>
<dc:creator>Tran, E.</dc:creator>
<dc:creator>Koh, A.</dc:creator>
<dc:creator>Lomeli, I.</dc:creator>
<dc:creator>Peretz, C. A. C.</dc:creator>
<dc:creator>Murad, N.</dc:creator>
<dc:creator>Abelson, S.</dc:creator>
<dc:creator>Morales, C.</dc:creator>
<dc:creator>Rivera, J. M.</dc:creator>
<dc:creator>Pikman, Y.</dc:creator>
<dc:creator>Cheng, M. L.</dc:creator>
<dc:creator>Logan, A. C.</dc:creator>
<dc:creator>Stieglitz, E.</dc:creator>
<dc:creator>Smith, C. C.</dc:creator>
<dc:date>2025-06-14</dc:date>
<dc:identifier>doi:10.1101/2025.06.10.658786</dc:identifier>
<dc:title><![CDATA[Multi-selective RAS(ON) Inhibition Targets Oncogenic RAS Mutations and Overcomes RAS/MAPK-Mediated Resistance to FLT3 and BCL2 Inhibitors in Acute Myeloid Leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.10.658982v1?rss=1">
<title>
<![CDATA[
A connectomic resource for neural cataloguing and circuit dissection of the larval zebrafish brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.10.658982v1?rss=1</link>
<description><![CDATA[
We present a correlated light and electron microscopy (CLEM) dataset from a 7-day-old larval zebrafish, integrating confocal imaging of genetically labeled excitatory (vglut2a) and inhibitory (gad1b) neurons with nanometer-resolution serial section EM. The dataset spans the brain and anterior spinal cord, capturing >180,000 segmented soma, >40,000 molecularly annotated neurons, and 30 million synapses, most of which were classified as excitatory, inhibitory, or modulatory. To characterize the directional flow of activity across the brain, we leverage the synaptic and cell body annotations to compute region-wise input and output drive indices at single cell resolution. We illustrate the datasets utility by dissecting and validating circuits in three distinct systems: water flow direction encoding in the lateral line, recurrent excitation and contralateral inhibition in a hindbrain motion integrator, and functionally relevant targeted long-range projections from a tegmental excitatory nucleus, demonstrating that this resource enables rigorous hypothesis testing as well as exploratory-driven circuit analysis. The dataset is integrated into an open-access platform optimized to facilitate community reconstruction and discovery efforts throughout the larval zebrafish brain.
]]></description>
<dc:creator>Petkova, M. D.</dc:creator>
<dc:creator>Januszewski, M.</dc:creator>
<dc:creator>Blakely, T.</dc:creator>
<dc:creator>Herrera, K. J.</dc:creator>
<dc:creator>Schuhknecht, G. F. P.</dc:creator>
<dc:creator>Tiller, R.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Schalek, R. L.</dc:creator>
<dc:creator>Boulanger-Weil, J.</dc:creator>
<dc:creator>Peleg, A.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Troidl, J.</dc:creator>
<dc:creator>Kumar Vohra, S.</dc:creator>
<dc:creator>Wei, D.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Bahl, A.</dc:creator>
<dc:creator>Tapia, J. C.</dc:creator>
<dc:creator>Iyer, N.</dc:creator>
<dc:creator>Miller, Z. T.</dc:creator>
<dc:creator>Hebert, K. B.</dc:creator>
<dc:creator>Pavarino, E. C.</dc:creator>
<dc:creator>Taylor, M.</dc:creator>
<dc:creator>Deng, Z.</dc:creator>
<dc:creator>Stingl, M.</dc:creator>
<dc:creator>Hockling, D.</dc:creator>
<dc:creator>Hebling, A.</dc:creator>
<dc:creator>Wang, R. C.</dc:creator>
<dc:creator>Zhang, L. L.</dc:creator>
<dc:creator>Dvorak, S.</dc:creator>
<dc:creator>Faik, Z.</dc:creator>
<dc:creator>King, K. I.</dc:creator>
<dc:creator>Goel, P.</dc:creator>
<dc:creator>Wagner-Carena, J.</dc:creator>
<dc:creator>Aley, D.</dc:creator>
<dc:creator>Chalyshkan, S.</dc:creator>
<dc:creator>Contreas, D.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Muthukumar, A. V.</dc:creator>
<dc:creator>Vernaglia, M. S.</dc:creator>
<dc:creator>Tapia Carrasco, T.</dc:creator>
<dc:creator>Melnychuck, S.</dc:creator>
<dc:creator>Yan, T.</dc:creator>
<dc:creator>Dalal, A.</dc:creator>
<dc:creator>DiMarti</dc:creator>
<dc:date>2025-06-15</dc:date>
<dc:identifier>doi:10.1101/2025.06.10.658982</dc:identifier>
<dc:title><![CDATA[A connectomic resource for neural cataloguing and circuit dissection of the larval zebrafish brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.11.659167v1?rss=1">
<title>
<![CDATA[
A lipid plug affects K2P6.1(TWIK-2) function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.11.659167v1?rss=1</link>
<description><![CDATA[
Lipids are integral to ion channel function yet delineating mechanisms by which they affect function remains challenging. Within the K2P family of leak potassium channels1-3, observation of tubular densities interpreted as alkyl chains occupying lateral fenestrations linking the pore and bilayer4-8 raised the possibility that lipid access from the bilayer acts as a regulatory mechanism4-7. Here, we present cryo-electron microscopy (cryo-EM) structures of the human leak potassium channel K2P6.1 (TWIK2)9-11 and mutants in nanodisc and detergent environments that reveal an unusual conformation in the first selectivity filter (SF1) and a pair of two-chain lipids within the channel cavity (denoted the  lipid plug). The chains of each plug lipid occupy separate binding sites that laterally extend to the bilayer from the channel cavity. One, the upper leg, matches the previously identified alkyl chain binding site4-8,12. The second, the lower leg, occupies a fenestration common with K2P1.1 (TWIK1)13. Together, they demonstrate a bidentate means to coordinate each plug lipid that offers a reinterpretation of previous observations. Structures of a K2P6.1 (TWIK2) mutant that directs the channel to the plasma membrane14 and an R257A mutant that increases function yield plugged and unplugged forms. Notably, the R257A plugged form shows a change in lipid plug position, indicating a key role for this residue in lipid binding. Together, our data suggest that occupation of the central cavity by the lipid plug serves as a mechanism to render the TWIK channels inactive and points to the importance of lipid plug removal to create an ion permeable pore. Such a mechanism could provide a potent way for limiting the leak function of K2Ps based on cellular location or other contextual factors.
]]></description>
<dc:creator>Mondal, A.</dc:creator>
<dc:creator>Niranjan, S.</dc:creator>
<dc:creator>Minor, D. L.</dc:creator>
<dc:date>2025-06-15</dc:date>
<dc:identifier>doi:10.1101/2025.06.11.659167</dc:identifier>
<dc:title><![CDATA[A lipid plug affects K2P6.1(TWIK-2) function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.13.659591v1?rss=1">
<title>
<![CDATA[
Increased Synapse Elimination by Inflammatory Cells Contributes to Long-lasting Post-Stroke Memory Dysfunction in Old Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.13.659591v1?rss=1</link>
<description><![CDATA[
Old patients are more likely to experience memory dysfunction than young patients after a stroke. It has been reported that brain astrocytes and microglia cause excessive removal of synapses at the acute and subacute stages of stroke, and inhibition of their phagocytosis improved neurobehavioral outcomes. We hypothesized that memory dysfunction in old subjects is associated with increased synapse removal by inflammatory cells. Ischemic stroke was induced in young (2-month-old) and old (15-18-month-old) mice. Memory functions were analyzed by the Y-maze test weekly for 8 weeks and the novel object recognition (NOR) test at 7 days before and 8 weeks post-stroke. We have also created a tibia fracture 6 hours before stroke injury in young mice, to test if the activation of 7-nicotinic acetylcholine receptor (nAchRs) reduces inflammatory cells and synapse elimination. Brains were collected 8 weeks after the induction of ischemic stroke. Transcriptome changes, neuronal injuries, neuroinflammation, synapse removal, and neurite outgrowth were analyzed. We found that old mice developed long-term memory dysfunction after ischemic stroke, which was not seen in young mice. Old mice showed larger infarct volume, higher neuroinflammation, and more synapses engulfed by microglia/macrophages and astrocytes in the peri-atrophic region and hippocampi than young mice. More synapse-engulfing astrocytes than microglia/macrophages were present in the peri-atrophic region and the ipsilateral hippocampi, suggesting that reactive astrocytes contributed more than activated microglia/macrophages in synapse removal. Activation of 7-nAchRs in mice subjected to tibia fracture 6 hours before ischemic injury reduced synapse removal by microglia/macrophages and astrocytes in the hippocampi. Our study indicated that an increase in synaptic elements by inflammatory cells contributes to the long-lasting memory deficit after stroke in old mice. Astrocytes may contribute more than microglia/macrophages in synapse removal. Inhibition of neuroinflammation by activating 7-nAchRs can reduce synapse loss and thus may improve post-stroke memory function.
]]></description>
<dc:creator>Shabani, Z.</dc:creator>
<dc:creator>Pan, P.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Leng, K.</dc:creator>
<dc:creator>Sangras, S.</dc:creator>
<dc:creator>Huo, K.</dc:creator>
<dc:creator>Yadav, A.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Shi, J.</dc:creator>
<dc:creator>Chinn, G.</dc:creator>
<dc:creator>Kyritsis, N.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Hellman, J.</dc:creator>
<dc:creator>Su, H.</dc:creator>
<dc:date>2025-06-15</dc:date>
<dc:identifier>doi:10.1101/2025.06.13.659591</dc:identifier>
<dc:title><![CDATA[Increased Synapse Elimination by Inflammatory Cells Contributes to Long-lasting Post-Stroke Memory Dysfunction in Old Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.12.659380v1?rss=1">
<title>
<![CDATA[
Rapid acquisition of HIV-1 neutralization breadth in a rhesus V2 apex germline antibody mouse model after a single bolus immunization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.12.659380v1?rss=1</link>
<description><![CDATA[
Current vaccine strategies to elicit broadly neutralizing antibodies (bnAbs) against HIV-1 generally propose complex, multi-boost immunization regimens. In rhesus macaques, SHIV infection has been observed to rapidly drive the development of some classes of bnAbs that share structural similarities with those in humans. Here, we generated a knockin mouse model with B cells bearing the unmutated common ancestor (UCA) of the V2 apex-targeted bnAb lineage, V033-a. A single immunization of mice with a germline-targeting native-like trimer was sufficient to recapitulate the ontogeny of the mature rhesus bnAb in knockin mice--including rare, disfavored somatic mutations--leading to the induction of antibodies that exhibited potent neutralization against both autologous and heterologous tier 2 viruses. A boost with Env escape mutant trimers further improved breadth and potency, and cryo-EM structure revealed the structural basis for heterologous neutralization breadth. Non-human primate and mouse models can thus combine with structure to serve as a platform for identifying and confirming immunogens that streamline HIV-vaccination regimens.
]]></description>
<dc:creator>Ghosh, A. R.</dc:creator>
<dc:creator>Habib, R.</dc:creator>
<dc:creator>Mishra, N.</dc:creator>
<dc:creator>Roark, R. S.</dc:creator>
<dc:creator>Akauliya, M.</dc:creator>
<dc:creator>Albowaidey, A. A.</dc:creator>
<dc:creator>Allen, J. D.</dc:creator>
<dc:creator>Amereh, K.</dc:creator>
<dc:creator>Avillion, G.</dc:creator>
<dc:creator>Bottermann, M.</dc:creator>
<dc:creator>Liang, B.</dc:creator>
<dc:creator>Chaudhary, N.</dc:creator>
<dc:creator>Callaghan, S.</dc:creator>
<dc:creator>Dye, J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Ellis-Pugh, J. R.</dc:creator>
<dc:creator>Chowdhury, R. R.</dc:creator>
<dc:creator>James, N. E.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Maiorino, L.</dc:creator>
<dc:creator>Villavicencio, P. M.</dc:creator>
<dc:creator>Nedellec, R.</dc:creator>
<dc:creator>Oberoi, P.</dc:creator>
<dc:creator>Sowers, K. J.</dc:creator>
<dc:creator>Park, Y.</dc:creator>
<dc:creator>Prum, T.</dc:creator>
<dc:creator>Rodriguez, L.</dc:creator>
<dc:creator>Ssozi, M.</dc:creator>
<dc:creator>Torres, J.</dc:creator>
<dc:creator>Walsh, A. A.</dc:creator>
<dc:creator>Warner, J. E.</dc:creator>
<dc:creator>Weldon, S. R.</dc:creator>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Wiehe, K.</dc:creator>
<dc:creator>Crispin, M.</dc:creator>
<dc:creator>Ward, A. B.</dc:creator>
<dc:creator>Nair, U.</dc:creator>
<dc:creator>Hahn, B. H.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Shapiro, L.</dc:creator>
<dc:creator>Kwong, P. D.</dc:creator>
<dc:creator>Irvine, D. J.</dc:creator>
<dc:creator>Andrabi, R.</dc:creator>
<dc:creator>Shaw, G. M.</dc:creator>
<dc:creator>Batista, F.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.12.659380</dc:identifier>
<dc:title><![CDATA[Rapid acquisition of HIV-1 neutralization breadth in a rhesus V2 apex germline antibody mouse model after a single bolus immunization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.16.660010v1?rss=1">
<title>
<![CDATA[
Genome editing of phylogenetically distinct bacteria using portable retron-mediated recombineering 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.16.660010v1?rss=1</link>
<description><![CDATA[
Advanced genome editing technologies have enabled rapid and flexible rewriting of the Escherichia coli genome, benefiting fundamental biology and biomanufacturing. Unfortunately, some of the most useful technologies to advance genome editing in E. coli have not yet been ported into other bacterial species. For instance, the addition of bacterial retrons to the genome editing toolbox has increased the efficiency of recombineering in E. coli by enabling sustained, abundant production of ssDNA recombineering donors by reverse transcription that install flexible, precise edits in the prokaryotic chromosome. To extend the utility of this technology beyond E. coli, we surveyed the portability and versatility of retron-mediated recombineering across three different bacterial phyla (Proteobacteria, Bacillota and Actinomycetota) and a total of 15 different species. We found that retron recombineering is functional in all species tested, reaching editing efficiencies above 20% in six of them, above 40% in three of them, and above 90% in two of them. We also tested the extension of the recombitron architecture optimizations and strain backgrounds in a subset of hosts to additionally increase editing rates. The broad recombitron survey carried out in this study forms the basis for widespread use of retron-derived technologies through the whole Bacteria domain.
]]></description>
<dc:creator>Gonzalez-Delgado, A.</dc:creator>
<dc:creator>Bonillo-Lopez, L.</dc:creator>
<dc:creator>Johnson, M. S.</dc:creator>
<dc:creator>Knoedlseder, N.</dc:creator>
<dc:creator>Ko, C.-C.</dc:creator>
<dc:creator>Lekbach, Y.</dc:creator>
<dc:creator>Oh, J.-H.</dc:creator>
<dc:creator>Selvakumar, H.</dc:creator>
<dc:creator>Wold, M. C.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Aragon, V.</dc:creator>
<dc:creator>Gralnick, J. A.</dc:creator>
<dc:creator>Guell, M.</dc:creator>
<dc:creator>Hatfull, G.</dc:creator>
<dc:creator>Keitz, B. K.</dc:creator>
<dc:creator>Koskella, B.</dc:creator>
<dc:creator>Mutalik, V. K.</dc:creator>
<dc:creator>van Pijkeren, J. P.</dc:creator>
<dc:creator>Shipman, S.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.16.660010</dc:identifier>
<dc:title><![CDATA[Genome editing of phylogenetically distinct bacteria using portable retron-mediated recombineering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.17.660221v1?rss=1">
<title>
<![CDATA[
FOXM1 Inhibition Promotes Polyploidization and Metabolic Maturation in Human iPSC-Derived Hepatocytes by Modulating the Wnt/β-Catenin Pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.17.660221v1?rss=1</link>
<description><![CDATA[
Human induced pluripotent stem cell (iPSCs)-derived hepatocytes are widely used in regenerative medicine and disease modeling. However, existing protocols mainly produce fetal-like cells, limiting accurate modeling of liver functionality. Topoisomerase II (TOP2) and its transcription factor, forkhead box M1 (FOXM1), are silenced during late liver embryonic development; however, their roles in hepatocyte differentiation remain unclear. Here, we examined the effects of TOP2 and FOXM1 inhibition on the terminal differentiation of hepatocytes. We found that subtoxic TOP2 inhibition reduced nuclear chromatin condensation without causing DNA damage. RNA-seq analysis showed that TOP2 inhibition induced cell cycle arrest in a TOP2A-selective manner, with FOXM1 downregulation. ATAC-seq validation demonstrated that TOP2A inhibition decreases chromatin accessibility and modulates the Wnt/{beta}-catenin pathway. Proteomic analysis revealed that FOXM1 inhibition modulated TOP2A expression, replicated TOP2A-mediated cell cycle arrest, and reduced the levels of fetal hepatocyte proteins (HBG1/2, UGT2B7, and AFP). Prolonged FOXM1 inhibition is correlated with increased hepatocyte polyploidization, enhanced CYP450 activity, and improved lipid metabolism, suggesting a potential role in these processes. Overall, our findings suggest that FOXM1 inhibition significantly promotes the terminal differentiation of human iPSC-derived hepatocytes, indicating a potential role for FOXM1 and TOP2A in liver development, regeneration, and disease.
]]></description>
<dc:creator>Telles-Silva, K. A.</dc:creator>
<dc:creator>Pacheco, L.</dc:creator>
<dc:creator>Komatsu, S.</dc:creator>
<dc:creator>Chianca, F.</dc:creator>
<dc:creator>Chagas, G.</dc:creator>
<dc:creator>Martins, G. C.</dc:creator>
<dc:creator>Gridina, M.</dc:creator>
<dc:creator>Panchenko, D.</dc:creator>
<dc:creator>Carvalho, V. M.</dc:creator>
<dc:creator>Caldini, E. G.</dc:creator>
<dc:creator>Fishman, V. S.</dc:creator>
<dc:creator>Arkin, M.</dc:creator>
<dc:creator>Goulart, E.</dc:creator>
<dc:creator>Zatz, M.</dc:creator>
<dc:date>2025-06-18</dc:date>
<dc:identifier>doi:10.1101/2025.06.17.660221</dc:identifier>
<dc:title><![CDATA[FOXM1 Inhibition Promotes Polyploidization and Metabolic Maturation in Human iPSC-Derived Hepatocytes by Modulating the Wnt/β-Catenin Pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.18.660470v1?rss=1">
<title>
<![CDATA[
Comprehensive Cross-Domain Taxonomic Classification of Microbiotas using Partitioned Amplification Multiplexed Amplicon Sequencing (PAMA-seq) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.18.660470v1?rss=1</link>
<description><![CDATA[
Microbial communities encompass diverse bacteria, archaea, and eukaryotes that play vital roles in ecosystems and host health. Comprehensive analysis of these communities requires accurate, quantitative, and cross-domain profiling, yet current sequencing methods--metagenome shotgun sequencing (MGS) and ribosomal RNA (rRNA) amplicon sequencing--struggle to achieve these goals in a single assay. MGS provides broad functional insights but suffers from high cost, computational burden, and reliance on incomplete reference databases. rRNA amplicon sequencing, while more taxonomically targeted, typically profiles either prokaryotes or eukaryotes separately, depending on the chosen primer sets, and thus lacks simultaneous cross-domain resolution. To address these limitations, we developed PAMA-seq, a droplet-digital multiplex PCR technique. PAMA-seq partitions DNA sample from microbial communities into nanoliter droplets, each containing a single template molecule, and independently amplifies both the 16S and 18S rRNA genes, resulting in uniform amplification efficiency and accurate quantification. We validated PAMA-seq on a synthetic microbial community, a stool sample from a patient with colorectal cancer, and a coastal seawater sample. The method provided stable, cross-domain taxonomic profiles at sequencing depths as low as 104 reads--one to two orders of magnitude fewer than comparable shotgun metagenomics approaches--while significantly reducing variability between replicates. By combining comprehensive domain coverage with low sequencing depth requirements, PAMA-seq offers an efficient, cost-effective, and scalable method for monitoring microbial communities across diverse ecosystems, from clinical samples to global marine environments.
]]></description>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Trepka, K. R.</dc:creator>
<dc:creator>Song, F.</dc:creator>
<dc:creator>Kyaw, T. S.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:creator>Abate, A. J.</dc:creator>
<dc:date>2025-06-19</dc:date>
<dc:identifier>doi:10.1101/2025.06.18.660470</dc:identifier>
<dc:title><![CDATA[Comprehensive Cross-Domain Taxonomic Classification of Microbiotas using Partitioned Amplification Multiplexed Amplicon Sequencing (PAMA-seq)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.14.659672v1?rss=1">
<title>
<![CDATA[
Induction of meningioma stem signature via hydrogel reprogramming and application of meningioma stem cell marker CXCR4 to pathological diagnosis and treatment. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.14.659672v1?rss=1</link>
<description><![CDATA[
BackgroundsMeningioma accounts for about 40% of brain tumors, but there was no effective treatment for recurrent or inoperable cases. We have previously found that when cancer cells are cultured on certain hydrogels, cancer stem cells are efficiently induced in various cancer types and we named this process as hydrogel activated reprogramming (HARP) phenomenon. In this study, we aimed to identify a key molecule that induced meningioma stem cells via hydrogels.

MethodsMeningioma cells cultured on hydrogels were analyzed for expression of regular stem cell markers and their tumor genericity. Microarray analysis was performed to identify meningioma stem cell specific markers and to examine the application of such marker molecules as a therapeutic targets or pathological diagnosis for grading.

ResultsRegular stem cell markers such as Nanog, and Oct3/4 were induced by culturing meningioma cells on hydrogels, and comprehensive gene expression analysis identified molecules involved in cancer stem cell activity. Among them, CXCR4 was selected as a therapeutic target molecule. Stimulation of CXCR4 via CXCL12 led to an increase in stem cell markers. In human meningioma pathological specimens and cultured cell lines, there was a correlation between CXCR4 expression levels and NF2 mutations and/or deletions. CXCR4 immunohistochemistry also detected along with the brain invasion. Thus, CXCR4 immunohistochemistry may be useful to suggest typical CNS WHO grade 1 meningioma, that do not require molecular analysis.

ConclusionsWe have defined meningioma stem cell signature via HARP phenomenon and identified CXCR4 with biological significance as being diagnostic target.

Key PointsO_LIWe induce meningioma stem signature via Hydrogel activated reprogramming (HARP) phenomena.
C_LIO_LIWe identified CXCR4 as a candidate molecule as stemness marker and therapeutic target.
C_LIO_LITranslational research using pathological specimens reveal CXCR4 immunohistochemistry has been shown to play an auxiliary role in the diagnosis of meningiomas.
C_LI

Importance of the StudyIn addition to morphology, immunohistochemistry and gene alteration are being adopted as diagnostic criteria for CNS tumors. From CNS5 onwards, epigenetic changes such as methylation analysis are being adopted as diagnostic criteria. We induced epigenetic changes in meningioma cells and induced cancer stemness, and this technique will be of great importance in the research and diagnosis of meningioma. In addition, microarray analysis was used to select CXCR4 from the molecules that increased simultaneously during stem cell induction on the three hydrogels, and further analysis revealed that CXCR4 immunostaining may indicate the distribution of meningioma stem cells, also making it useful for diagnosis. This report will contribute to the advancement of meningioma research and diagnosis by conducting basic research and using histopathological specimens.
]]></description>
<dc:creator>Oda, Y.</dc:creator>
<dc:creator>Tsuda, M.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Suzuka, J.</dc:creator>
<dc:creator>Yuzawa, S.</dc:creator>
<dc:creator>Ise, K.</dc:creator>
<dc:creator>Habiba, U.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Tanikawa, S.</dc:creator>
<dc:creator>Sugino, H.</dc:creator>
<dc:creator>Tanei, Z.-i.</dc:creator>
<dc:creator>Mawrin, C.</dc:creator>
<dc:creator>Gong, J. P.</dc:creator>
<dc:creator>Tanaka, S.</dc:creator>
<dc:date>2025-06-20</dc:date>
<dc:identifier>doi:10.1101/2025.06.14.659672</dc:identifier>
<dc:title><![CDATA[Induction of meningioma stem signature via hydrogel reprogramming and application of meningioma stem cell marker CXCR4 to pathological diagnosis and treatment.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.15.659784v1?rss=1">
<title>
<![CDATA[
Transient Dietary Changes Modulate Inflammatory Disease Trajectory in the Lean State 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.15.659784v1?rss=1</link>
<description><![CDATA[
Obesity potently alters immune responses across various inflammatory contexts1-7. However, it is unclear whether a transient high-fat diet (HFD) can affect immune function despite minimal effects on body weight. Here, we demonstrate that a short-term HFD regimen significantly exacerbates disease severity in a model of experimental psoriasis, comparable to what is observed in obese mice on long-term HFD. We find that a critical 4-day window of HFD coinciding with disease onset is sufficient to overactivate the immune response, leading to worsened disease outcomes. Mechanistically, we identify the CD4+ T cell compartment as an essential mediator of HFD-induced disease severity. Within the compartment, we functionally validate that disease exacerbation is driven by increased differentiation of a pathogenic population of T helper 17 (TH17) cells8 expressing IL1R1 (interleukin-1 receptor, type I)9, which is triggered by localized activation of the NLRP3 (NOD-, LRR-, and pyrin domain-containing protein 3) inflammasome10. This brief window of HFD at disease onset leads to immune-dependent sensitization, as disease severity is increased upon a subsequent flare, despite complete recovery and under a low-fat diet (LFD) regimen. Our data indicate that transient dietary changes during the initial stages of an inflammatory event can rewire the immune milieu, profoundly influencing disease progression and inflammatory memory11. This phenomenon may have broad implications for conditions where obesity, and not diet itself, is considered the primary risk factor.
]]></description>
<dc:creator>Jiang, Z.</dc:creator>
<dc:creator>Tayeri, J. M.</dc:creator>
<dc:creator>Tabuchi, C.</dc:creator>
<dc:creator>Gayer, S. G.</dc:creator>
<dc:creator>Yoo, A.</dc:creator>
<dc:creator>Sun, I.-H.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Yin, H.</dc:creator>
<dc:creator>Chuah, S. S.</dc:creator>
<dc:creator>Deshpande, P.</dc:creator>
<dc:creator>Johri, V.</dc:creator>
<dc:creator>Davidson, B.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Kuhn, N. F.</dc:creator>
<dc:creator>Courau, T.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Bapat, S. P.</dc:creator>
<dc:date>2025-06-20</dc:date>
<dc:identifier>doi:10.1101/2025.06.15.659784</dc:identifier>
<dc:title><![CDATA[Transient Dietary Changes Modulate Inflammatory Disease Trajectory in the Lean State]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.19.657327v1?rss=1">
<title>
<![CDATA[
Satb1 integrates cohesin mediated genome organization and transcriptional regulation during T cell development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.19.657327v1?rss=1</link>
<description><![CDATA[
Three-dimensional (3D) genome folding, which is highly cell type-specific, plays a crucial role in orchestrating spatiotemporal gene expression. Although factors such as CTCF have been extensively studied in the hierarchical regulation of 3D chromatin organization, the mechanisms driving dynamic genome folding during T cell fate transitions remain incompletely defined. In this study, we reveal that Satb1, a chromatin organizer enriched in the T cell lineage, co-occupies genomic regions with the cohesin complex and Ctcf in double-positive (DP) thymocytes, where chromatin interactions are notably increased. We show that Satb1 physically interacts with the cohesin subunit Smc1a, and its deletion results in aberrant Smc1a binding and reduced chromatin contacts at sites co-occupied by Satb1 and cohesin. In both DP and immature CD4 single-positive (SP) T cells, Satb1 is essential for proper T cell activation and cytokine signaling. At the Cd3 locus, Satb1 and cohesin collaboratively regulate gene expression, with Satb1 loss leading to disrupted Smc1a occupancy and compromised chromatin interactions. Furthermore, Satb1 shows in vitro properties consistent with liquid-liquid phase separation, and disease-associated mutations impair these properties. Together, our findings uncover a molecular mechanism in which Satb1 facilitates chromatin looping through direct interaction with the cohesin complex and its ability to form nuclear condensates, thereby governing transcriptional regulation during T cell development.
]]></description>
<dc:creator>Madhok, A.</dc:creator>
<dc:creator>Patta, I.</dc:creator>
<dc:creator>Desai, I.</dc:creator>
<dc:creator>Tayyab, M.</dc:creator>
<dc:creator>Choudhary, V.</dc:creator>
<dc:creator>Vishwakarma, S.</dc:creator>
<dc:creator>Saha, S.</dc:creator>
<dc:creator>Lahesmaa, R.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:creator>Galande, S.</dc:creator>
<dc:date>2025-06-20</dc:date>
<dc:identifier>doi:10.1101/2025.06.19.657327</dc:identifier>
<dc:title><![CDATA[Satb1 integrates cohesin mediated genome organization and transcriptional regulation during T cell development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.19.660622v1?rss=1">
<title>
<![CDATA[
ADAR1 Restricts Poliovirus Infection through dual disruption of translation initiation and protein coding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.19.660622v1?rss=1</link>
<description><![CDATA[
Viral RNAs (vRNAs) interact with hundreds of host proteins, but how these interactions shape viral infection remains largely unknown. Here we developed a mass-spectrometry approach to identify cellular RNA-binding proteins (RBPs) that enhance or inhibit infection by modulating vRNA translation. We identified 130 RBPs that associate with polysomes in human cells infected with poliovirus, including known regulators of cap-independent translation from the viral internal ribosome entry site (IRES). We find that adenosine deaminase acting on RNA (ADAR1) is recruited to polysomes during infection and edits 3 sites in the vRNA, reducing viral replication. Two edits occur in the IRES and impair internal translation initiation, while the third edit occurs in the coding region and triggers an amino acid substitution. Incorporation of this threonine-to-alanine mutation into the viral genome attenuated replication. We conclude that ADAR1 restricts poliovirus infection by both reducing vRNA translation and introducing a recurrent coding error.
]]></description>
<dc:creator>Kim, D.</dc:creator>
<dc:creator>McCarthy, F.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Chalkley, R. J.</dc:creator>
<dc:creator>Pak, J.</dc:creator>
<dc:creator>Elias, J. E.</dc:creator>
<dc:creator>Aviner, R.</dc:creator>
<dc:date>2025-06-23</dc:date>
<dc:identifier>doi:10.1101/2025.06.19.660622</dc:identifier>
<dc:title><![CDATA[ADAR1 Restricts Poliovirus Infection through dual disruption of translation initiation and protein coding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.21.660874v1?rss=1">
<title>
<![CDATA[
Control of Quiescence and Activation of Human Muscle Stem Cells by Cytokines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.21.660874v1?rss=1</link>
<description><![CDATA[
Skeletal muscle homeostasis and repair depend on the activation of tissue resident stem cells called satellite cells. To understand the early molecular basis of human satellite cell activation, epigenomics, transcriptomics and protein analysis were performed in quiescent and activated human satellite cells. Cytokine signaling pathways were enriched in stimulated human satellite cells revealing a high cytokine enrichment, including CCL2, CCL20, CXCL8, IL-6, TNFRSF12A, ILR1, CSF-1 and FGF2. Functional roles of these observed changes are supported by in vivo experiments showing that chemokine inhibitors increase engraftment and regeneration capacity of human satellite cells xenotransplants. Cytokines, chemokines and associated signaling pathways in the early stages of human satellite cell activation may underlie disparate muscle responses in neuromuscular inflammatory and degenerative disorders and consequently are potential entry points for clinical applications towards muscle repair.
]]></description>
<dc:creator>Striedinger, K.</dc:creator>
<dc:creator>Barruet, E.</dc:creator>
<dc:creator>Atamaniauc, E.</dc:creator>
<dc:creator>Linquist, K.</dc:creator>
<dc:creator>Knott, C.</dc:creator>
<dc:creator>Brack, A.</dc:creator>
<dc:creator>Pomerantz, J. H.</dc:creator>
<dc:date>2025-06-23</dc:date>
<dc:identifier>doi:10.1101/2025.06.21.660874</dc:identifier>
<dc:title><![CDATA[Control of Quiescence and Activation of Human Muscle Stem Cells by Cytokines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.18.659743v1?rss=1">
<title>
<![CDATA[
Developmental Plasticity and Stromal Co-option Shape a Pituitary Neuroendocrine Tumor Transcriptional Continuum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.18.659743v1?rss=1</link>
<description><![CDATA[
Pituitary neuroendocrine tumors (PitNETs) are common intracranial neoplasms with complex biology underpinned by unresolved cellular origins, molecular heterogeneity, and microenvironment interactions. Here, we employ single-nuclei RNA-sequencing (snRNA-seq) of 419,874 cells from human normal pituitaries and PitNETs with spatial transcriptomics to resolve these challenges. We identify multi-hormonal neuroendocrine cells in both normal and tumor tissues, originating as early pseudotime intermediates from pituitary stem cells, revealing an inherent plasticity that blurs traditional lineage boundaries. PitNETs exhibit a transcriptional continuum across subtypes, challenging their classification into discrete categories. Trajectory analysis uncovers divergent cellular origins: silent gonadotroph adenomas (SGAs), prolactinomas, silent corticotroph adenomas (SCAs), and Cushings adenomas are closely linked to differentiated neuroendocrine cells, while somatotroph and null cell adenomas (NCAs) appear to derive more directly from adult stem cells. Tumor cells co-opt robust cell-cell communication networks found in normal adult neuroendocrine cells. Spatial profiling further demonstrates that perivascular niches enhance tumorigenicity through angiogenic and epithelial-mesenchymal transition programs. Our work redefines PitNETs as ecosystems shaped by developmental plasticity and microenvironmental crosstalk, offering a roadmap for future therapies targeting lineage fluidity and stromal dependencies.
]]></description>
<dc:creator>Osorio, R. C.</dc:creator>
<dc:creator>Oh, J. Y.</dc:creator>
<dc:creator>Jung, J.</dc:creator>
<dc:creator>Ehrenberg, A. J.</dc:creator>
<dc:creator>Saha, A.</dc:creator>
<dc:creator>Lad, M.</dc:creator>
<dc:creator>Khela, H.</dc:creator>
<dc:creator>Brennick, N.</dc:creator>
<dc:creator>Giannikopoulos, P.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Diaz, A.</dc:creator>
<dc:creator>Aghi, M. K.</dc:creator>
<dc:date>2025-06-24</dc:date>
<dc:identifier>doi:10.1101/2025.06.18.659743</dc:identifier>
<dc:title><![CDATA[Developmental Plasticity and Stromal Co-option Shape a Pituitary Neuroendocrine Tumor Transcriptional Continuum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.18.660471v1?rss=1">
<title>
<![CDATA[
Improving the Scalability of Bayesian Phylodynamic Inference through Efficient MCMC Proposals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.18.660471v1?rss=1</link>
<description><![CDATA[
In Bayesian phylodynamics, we jointly reconstruct the posterior distribution of timed phylogenetic trees, evolutionary and population dynamic parameters. Most approaches in the field use Markov chain Monte Carlo (MCMC) for inference. Inferring timed phylogenetic trees using MCMC relies on a set of proposal distributions - operators - to explore different phylogenetic tree topologies, node ranks, and heights. However, inefficiencies in these proposal distributions limit the speed and scale with which phylodynamic analyses can be performed. Here, we introduce two classes of operators that allow scaling of phylodynamic analyses by reducing the time needed to reach convergence. We propose an improved proposal distribution to scale trees based on the intervals between consecutive nodes in the tree, and a set of operators informed by a pseudo parsimony score to explore different tree topologies. Contrary to already existing operators that make large changes to areas of the tree with no mutations at all, our new parsimony-focused operators target areas of the tree with many mutations.

We first demonstrate the correctness of these proposal distributions using a well-calibrated simulation study. We then demonstrate the improvements of using the updated proposal distributions using a number of large real-world datasets, showing a substantial increase in effective sample size per hour across viral datasets as well as a reduction in the number of samples required for the MCMC chain to reach stationarity. Finally, we apply the operators to a 10,000-sequence H3N2 dataset including phylogeography and finish the analysis within two weeks. The operators are implemented in the TargetedBeast package, which is available as an open-source package for BEAST2 and a user-friendly graphical user interface. The speed improvements in the TargetedBeast package are beneficial to any phylodynamics method implemented in BEAST2 that relies on standard phylogenetic trees, such as coalescent-skyline, birth-death skyline, and structured phylodynamic methods.
]]></description>
<dc:creator>Bouckaert, R.</dc:creator>
<dc:creator>Weidemüller, P.</dc:creator>
<dc:creator>Esquivel Gomez, L.</dc:creator>
<dc:creator>Müller, N. F.</dc:creator>
<dc:date>2025-06-24</dc:date>
<dc:identifier>doi:10.1101/2025.06.18.660471</dc:identifier>
<dc:title><![CDATA[Improving the Scalability of Bayesian Phylodynamic Inference through Efficient MCMC Proposals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.23.661129v1?rss=1">
<title>
<![CDATA[
Peroxisomal import is circadian in glia and regulates sleep and lipid metabolism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.23.661129v1?rss=1</link>
<description><![CDATA[
Peroxisomes are critical organelles that detoxify cellular waste while also catabolizing and anabolizing lipids. How peroxisomes coordinate protein import and support metabolic functions across complex tissues and timescales remains poorly understood in vivo. Using the Drosophila brain, we discover a striking enrichment of peroxisomes in the neuronal soma and the cortex glia that enwrap them. Unexpectedly, import of peroxisomal proteins into cortex glia, but not neurons, oscillated across time and peaked in the early morning. Rhythmic peroxisomal import in cortex glia autonomously required the circadian clock and Peroxin 5 (Pex5; peroxisomal biogenesis factor 5 homolog), with import persistently elevated in clock mutants. Notably, reducing Pex5 in cortex glia, but not neurons, caused hyperactivity and reduced total sleep. Moreover, brain lipid metabolism was dramatically altered upon Pex5 knockdown, with glia impacting sphingolipids and triacylglycerols, and neurons impacting phospholipids. The cell-type specificity of these Pex5 phenotypes highlights unique roles for peroxisomal import in both sleep and lipid metabolism in the brain.
]]></description>
<dc:creator>Das, A.</dc:creator>
<dc:creator>Serna, I. M. R.</dc:creator>
<dc:creator>Kumar, A.</dc:creator>
<dc:creator>Kalita, H.</dc:creator>
<dc:creator>Sherpa, L.</dc:creator>
<dc:creator>Vaughen, J. P.</dc:creator>
<dc:creator>Bai, H.</dc:creator>
<dc:date>2025-06-25</dc:date>
<dc:identifier>doi:10.1101/2025.06.23.661129</dc:identifier>
<dc:title><![CDATA[Peroxisomal import is circadian in glia and regulates sleep and lipid metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.24.661339v1?rss=1">
<title>
<![CDATA[
Plasma Proteomic Profiling Reveals Distinct Signatures of Chest CT Phenotypes in Sarcoidosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.24.661339v1?rss=1</link>
<description><![CDATA[
BackgroundSarcoidosis is a granulomatous disease of unknown cause with a highly variable clinical course. The inability to predict progressive inflammation, fibrosis, or both underscores the limited understanding of the underlying molecular mechanisms.

ObjectiveWe aimed to identify novel protein signatures associated with distinct pulmonary phenotypes of sarcoidosis, including progressive inflammation, progressive fibrosis, and disease resolution.

MethodsWe performed the SomaScan 11K Assay to measure more than 10,000 unique human plasma proteins and compared protein expression between chest CT-defined phenotypes using principal component analysis, differential expression, correlation analysis, and gene set enrichment analysis.

ResultsWe identified distinct proteomic signatures that differentiate progressive fibrosis from progressive nodular inflammation in sarcoidosis. Enrichment and differential expression analyses revealed that progressive fibrosis was associated with epithelial-mesenchymal transition pathways, while progressive nodular disease was linked to mTORC1 and MYC signaling, as well as metabolic activation. Additionally, expression of 44 proteins correlated moderately to strongly with thoracic lymph node enlargement, suggesting that lymph node- driven immune activity may be a major source of circulating proteomic signals.

ConclusionsThis study leverages a unique longitudinal imaging approach to define extreme pulmonary phenotypes based on serial chest CT scoring, enabling the discovery of proteomic signals linked to distinct trajectories of sarcoidosis progression. Once validated, these findings could inform the development of blood-based biomarkers for disease stratification, monitoring, and therapeutic targeting in sarcoidosis.
]]></description>
<dc:creator>Shastry, V.</dc:creator>
<dc:creator>Leach, S. M.</dc:creator>
<dc:creator>Elicker, B. M.</dc:creator>
<dc:creator>Koth, L. L.</dc:creator>
<dc:date>2025-06-25</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.661339</dc:identifier>
<dc:title><![CDATA[Plasma Proteomic Profiling Reveals Distinct Signatures of Chest CT Phenotypes in Sarcoidosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.24.661379v1?rss=1">
<title>
<![CDATA[
De novo designed bright, hyperstable rhodamine binders for fluorescence microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.24.661379v1?rss=1</link>
<description><![CDATA[
De novo protein design has emerged as a powerful strategy with the promise to create new tools. The practical performance of designed fluorophore binders, however, has remained far from meeting fluorescence microscopy demands. Here, we design de novo Rhodamine Binder (Rhobin) tags that combine ideal properties including size, brightness, and now adding hyperstability. Rhobin allows live and fixed cell imaging of a wide range of subcellular targets in mammalian cells. Its reversible fluorophore binding further enables live super-resolution STED microscopy with low photobleaching, as well as PAINT-type single-molecule localization microscopy. We showcase Rhobin in the extremophile Sulfolobus acidocaldarius living at 75{degrees}C, an application previously inaccessible by existing tags. Rhobin will serve as the basis for a new class of live cell fluorescent tags and biosensors.
]]></description>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Yserentant, K.</dc:creator>
<dc:creator>Hong, K.</dc:creator>
<dc:creator>Kuang, Y.</dc:creator>
<dc:creator>Bhowmick, A.</dc:creator>
<dc:creator>Charles-Orszag, A.</dc:creator>
<dc:creator>Lord, S. J.</dc:creator>
<dc:creator>Lu, L.</dc:creator>
<dc:creator>Hou, K.</dc:creator>
<dc:creator>Mann, S. I.</dc:creator>
<dc:creator>Grimm, J. B.</dc:creator>
<dc:creator>Lavis, L. D.</dc:creator>
<dc:creator>Mullins, R. D.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:date>2025-06-25</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.661379</dc:identifier>
<dc:title><![CDATA[De novo designed bright, hyperstable rhodamine binders for fluorescence microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.26.661761v1?rss=1">
<title>
<![CDATA[
Early transcriptional signatures of MeCP2 positive and negative cells in Rett syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.26.661761v1?rss=1</link>
<description><![CDATA[
Rett syndrome (RTT) is an X-linked neurological disorder caused by MECP2 mutations. Like other X-linked disorders, RTT patients have sex-specific differences in clinical presentation due to distinct cellular environments, where females have [~]50% of cells expressing either a mutant or wild-type copy of MECP2 (mosaic) and males have 100% of cells expressing a mutant MECP2 (non-mosaic). Typical RTT females have a short window of normal early development until [~]6-18 months, followed by regression and progressive decline, whereas neonatal encephalopathy is more likely in RTT males. How these sex-specific differences in cellular context contribute molecularly to RTT pathogenesis, particularly in the presymptomatic stages of RTT females, remains poorly understood. Here, we profiled the hippocampal transcriptomes of female (Mecp2+/-) and male (Mecp2-/y) RTT mice at early timepoints using both bulk and single-nucleus RNA-seq, including sorted MeCP2 positive (MeCP2+) and MeCP2 negative (MeCP2-) neurons in female mice. We identified a core disease signature consisting of 12 genes consistently dysregulated only in MeCP2-cells across RTT models. Moreover, we uncovered non-cell-autonomous effects exclusively in female MeCP2+ excitatory neurons, but not inhibitory neurons, suggesting excitatory circuits are more vulnerable early in the mosaic RTT environment. The single-nuclei data also revealed that a previously underappreciated MeCP2-interneuron subtype had the most transcriptional dysregulation in both male and female RTT hippocampi. Together, these data highlight the different effects of MeCP2 loss on excitatory and inhibitory circuits between the mosaic and non-mosaic environment that appear early in RTT pathogenesis.
]]></description>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Anderson, A. G.</dc:creator>
<dc:creator>Qi, G.</dc:creator>
<dc:creator>Wu, S.-R.</dc:creator>
<dc:creator>Revelli, J.-P.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Zoghbi, H. Y.</dc:creator>
<dc:date>2025-06-26</dc:date>
<dc:identifier>doi:10.1101/2025.06.26.661761</dc:identifier>
<dc:title><![CDATA[Early transcriptional signatures of MeCP2 positive and negative cells in Rett syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.23.661120v1?rss=1">
<title>
<![CDATA[
Sensitizing Immune-Refractory Ovarian Tumors via p53 Mutation-Tailored Immunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.23.661120v1?rss=1</link>
<description><![CDATA[
High-grade serous ovarian cancer demonstrates limited responsiveness to immune checkpoint inhibitors, owing in part to immunosuppressive environments shaped by nearly universal p53 aberrations. Utilizing an immunocompetent mouse model and individual p53 mutations, we identified a dependence of the p53-R270H mutation (equivalent of human R273H) on regulatory T cells (Tregs) and the PD-1/PD-L1 axis. Analysis of patient datasets associated R273H with elevated levels of two p53 targets, PD-L1 and amphiregulin (AREG), a Tregs growth factor. In contrast to p53-R172H tumors, where there was limited activity, dual antibody therapy targeting AREG and PD-L1 selectively and effectively inhibited R270H tumors. This involved polarization toward M1 macrophages, infiltration of CD8+ T cells, diminished Ly6G+ neutrophils and downregulation of interleukin-4. In patient-derived R273C organoids, the combination treatment reduced the CD4/CD8 ratio. This study is the first to establish a mutation-tailored therapeutic approach that leverages the capacity of p53 to modulate immunosuppressive mechanisms.
]]></description>
<dc:creator>Chatterjee, R.</dc:creator>
<dc:creator>Simoni-Nieves, A.</dc:creator>
<dc:creator>Truong, A.</dc:creator>
<dc:creator>Lindzen, M.</dc:creator>
<dc:creator>Ozmen, F.</dc:creator>
<dc:creator>Cherry, C.</dc:creator>
<dc:creator>Zwicky, P.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Selvadurai, B.-R.</dc:creator>
<dc:creator>Salame, T. M.</dc:creator>
<dc:creator>Gupta, N.</dc:creator>
<dc:creator>Giri, S.</dc:creator>
<dc:creator>Kramarski, L.</dc:creator>
<dc:creator>Avraham, Y.</dc:creator>
<dc:creator>Weizman, E.</dc:creator>
<dc:creator>Ozmen, T.</dc:creator>
<dc:creator>Noronha, A.</dc:creator>
<dc:creator>Chakrabarti, P.</dc:creator>
<dc:creator>Ramesh-Kumar, D.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Dahan, R.</dc:creator>
<dc:creator>Amit, I.</dc:creator>
<dc:creator>Velculescu, V.</dc:creator>
<dc:creator>Brenton, J.</dc:creator>
<dc:creator>Mills, G.</dc:creator>
<dc:creator>Oren, M.</dc:creator>
<dc:creator>Yarden, Y.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.23.661120</dc:identifier>
<dc:title><![CDATA[Sensitizing Immune-Refractory Ovarian Tumors via p53 Mutation-Tailored Immunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.24.661128v1?rss=1">
<title>
<![CDATA[
Functionalizing hydrogel nanovials with vesicles mimicking antigen-presenting vesicles and cancer exosomes improves T cell capture and activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.24.661128v1?rss=1</link>
<description><![CDATA[
Recent advances have demonstrated the application of microcavity-containing hydrogel microparticles, known as nanovials, for the massively parallel and high-throughput screening of therapeutic T cell populations for adoptive cell therapies. Nanovial cavities coated with peptide-MHC (pMHC) or antigen tetramers selectively bind to their cognate T cell receptor (TCR) or chimeric antigen receptor (CAR) to activate T cells and capture secreted cytokines. However, binding of tetramers or recombinantly expressed antigen by T cells is not always correlated with T cell activation or cytotoxicity as the binding interface is not fully representative of the natural immunological synapse formed between T cells and professional antigen-presenting cells (APCs). Here, we leverage the recent discovery of an ESCRT- and ALIX-binding region (EABR) sequence to generate antigen-presenting vesicles and cancer-mimicking exosomes from standard HEK293T and Expi293F cell cultures. EABR-mediated vesicles present natural, full-length oncologically-relevant membrane proteins embedded in lipid bilayers to functionalize the nanovial cavity with cell-like membranes. These hydrogel nanovials functionalized with the EABR-mediated vesicles show improved T cell capture of 1G4 T cells and enhanced activation of HER2 CAR-T cells compared to hydrogel surfaces functionalized with recombinantly-expressed soluble proteins.
]]></description>
<dc:creator>Olson, B. A.</dc:creator>
<dc:creator>Mellody, M. P.</dc:creator>
<dc:creator>Soemardy, C.</dc:creator>
<dc:creator>Mao, Z.</dc:creator>
<dc:creator>Mei, A.</dc:creator>
<dc:creator>Lippert, K.</dc:creator>
<dc:creator>Hoffmann, M. A. G.</dc:creator>
<dc:creator>Di Carlo, D.</dc:creator>
<dc:creator>Mayo, S. L.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.661128</dc:identifier>
<dc:title><![CDATA[Functionalizing hydrogel nanovials with vesicles mimicking antigen-presenting vesicles and cancer exosomes improves T cell capture and activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.24.661380v1?rss=1">
<title>
<![CDATA[
Accurate variant effect estimation in FACS-based deep mutational scanning data with Lilace 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.24.661380v1?rss=1</link>
<description><![CDATA[
Deep mutational scanning (DMS) experiments interrogate the effect of genetic variants on protein function, often using fluorescence-activated cell sorting (FACS) to quantitatively measure molecular phenotypes, such as abundance or activity. Analysis of DMS experiments with a FACS readout is challenging due to measurement variance and the unique multidimensional nature of the phenotype. However, no statistical method has yet been developed to address the challenges of FACS-based DMS. Here we present Lilace, a Bayesian statistical model to estimate variant effects with uncertainty quantification from FACS-based DMS experiments. We validate Lilaces performance and robustness using simulated data and apply it to OCT1 and Kir2.1 DMS experiments, demonstrating an improved false discovery rate (FDR) while largely maintaining sensitivity.
]]></description>
<dc:creator>Freudenberg, J.</dc:creator>
<dc:creator>Rao, J.</dc:creator>
<dc:creator>Howard, M. K.</dc:creator>
<dc:creator>Macdonald, C.</dc:creator>
<dc:creator>Greenwald, N. F.</dc:creator>
<dc:creator>Coyote-Maestas, W.</dc:creator>
<dc:creator>Pimentel, H.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.661380</dc:identifier>
<dc:title><![CDATA[Accurate variant effect estimation in FACS-based deep mutational scanning data with Lilace]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.25.661602v1?rss=1">
<title>
<![CDATA[
A cell-based scrambling assay reveals phospholipid headgroup preference of TMEM16F on the plasma membrane 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.25.661602v1?rss=1</link>
<description><![CDATA[
The asymmetric resting distribution of the three major phospholipid classes on the mammalian plasma membrane, with phosphatidylserine and phosphatidylethanolamine mostly on the inner leaflet, and phosphatidylcholine mostly on the outer leaflet, is maintained by ATP-dependent flippases and floppases that exhibit headgroup selectivity. Upon signaling cues, this asymmetry can be dissipated by various phospholipid scramblases, allowing cells to respond to stimuli and adapt to different physiological contexts. The prevailing view in the field is that phospholipid scramblases on the plasma membrane act without headgroup preference. Here we report contrary experimental evidence based on a phospholipid scrambling assay that quantifies the fluorescence polarization of nitrobenzoxadiazole-labeled phospholipids for kinetic monitoring of phospholipid scrambling on the plasma membrane of living cells. Our experiments reveal that the plasma membrane-residing calcium-activated phospholipid scramblase TMEM16F preferentially acts on phosphatidylserine and phosphatidylcholine over phosphatidylethanolamine.

Significance StatementPhospholipid scramblases on the mammalian plasma membrane are thought to act promiscuously without preference for headgroup. Thoroughly addressing this question, however, requires the development of new methodologies. We devised a cell-based phospholipid scrambling assay that utilizes the fluorescence polarization of nitrobenzoxadiazole (NBD)-labeled phospholipids, allowing the monitoring of their scrambling in a native environment. We discovered that the plasma membrane-residing calcium-activated phospholipid scramblase TMEM16F preferentially acts on phosphatidylserine and phosphatidylcholine over phosphatidylethanolamine.

Major Classification: Biological Sciences; Minor Classification: Cell biology, Biophysics and Computational Biology
]]></description>
<dc:creator>Teo, C. F.</dc:creator>
<dc:creator>Tuomivaara, S. T.</dc:creator>
<dc:creator>van Hilten, N.</dc:creator>
<dc:creator>Crottes, D.</dc:creator>
<dc:creator>Jan, Y. N.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>Jan, L. Y.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.25.661602</dc:identifier>
<dc:title><![CDATA[A cell-based scrambling assay reveals phospholipid headgroup preference of TMEM16F on the plasma membrane]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.27.662040v1?rss=1">
<title>
<![CDATA[
Synaptic plasticity of prefrontal long-range inhibition regulates cognitive flexibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.27.662040v1?rss=1</link>
<description><![CDATA[
While glutamatergic synaptic plasticity is believed to be a fundamental mechanism mediating learning, the behavioral significance of plasticity at cortical GABAergic synapses remains less well understood. Furthermore, despite recent discoveries of long-range projections from neocortical GABAergic neurons, details about how they function are also sparse. Here we combine behavioral optogenetics with patch-clamp electrophysiology to link plasticity at long-range GABAergic synapses with higher-order cognitive functions. Specifically, learning extradimensional rule shifts potentiates callosal GABAergic synapses from prefrontal parvalbumin-expressing (PV) neurons onto corticothalamic neurons. Disrupting this potentiation by inhibiting callosal PV terminals during rule shifts induces perseveration, whereas reinstating this potentiation with subsequent gamma-frequency callosal PV terminal stimulation restores flexible behavior. This shows how a novel plasticity locus can regulate brain circuits underlying normal cognition and pathological states.
]]></description>
<dc:creator>Zhu, X.</dc:creator>
<dc:creator>Hagopian, L. L.</dc:creator>
<dc:creator>Wallquist, K. E.</dc:creator>
<dc:creator>Sohal, V. S.</dc:creator>
<dc:date>2025-06-28</dc:date>
<dc:identifier>doi:10.1101/2025.06.27.662040</dc:identifier>
<dc:title><![CDATA[Synaptic plasticity of prefrontal long-range inhibition regulates cognitive flexibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.25.661551v1?rss=1">
<title>
<![CDATA[
Dietary restriction promotes neuronal resilience via ADIOL 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.25.661551v1?rss=1</link>
<description><![CDATA[
The steroid hormone 5-androstene-3{beta},17{beta}-diol (ADIOL) was discovered in humans nearly a century ago, yet its physiological roles remain poorly defined. Here, we show that fasting and caloric restriction, two forms of dietary restriction, induce transcriptional upregulation of genes encoding CYP11A1, CYP17A1, and 17{beta}-hydroxysteroid dehydrogenase family enzymes, promoting ADIOL biosynthesis. ADIOL, in turn, acts on the nervous system to reduce levels of kynurenic acid, a neuroactive metabolite linked to cognitive decline and neurodegeneration. This effect requires NHR-91, the C. elegans homolog of estrogen receptor {beta}, specifically in the RIM neuron, a key site of kynurenic acid production. Consistent with the known benefits of fasting and caloric restriction on healthspan, enhancing ADIOL signaling improves multiple healthspan indicators during aging. Conversely, animals deficient in ADIOL signaling exhibit reduced healthspan under normal conditions and in genetic models of caloric restriction, underscoring the functional significance of this pathway. Notably, ADIOL does not significantly impact lifespan, indicating that its healthspan benefits are not simply a byproduct of lifespan extension. Together, these findings establish a physiological role for ADIOL in mediating the neuroprotective and pro-healthspan effects of fasting and caloric restriction and suggest that boosting ADIOL signaling may help narrow the gap between lifespan and healthspan. This positions ADIOL as a promising mimetic of dietary restriction effects on healthspan that could be used as a therapeutic strategy for age-related neurodegenerative conditions.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=96 SRC="FIGDIR/small/661551v2_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@3246f7org.highwire.dtl.DTLVardef@d13dbaorg.highwire.dtl.DTLVardef@12b86f6org.highwire.dtl.DTLVardef@a98d62_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Guijarro-Hernandez, A.</dc:creator>
<dc:creator>Yoo, S.</dc:creator>
<dc:creator>Lemieux, G. A.</dc:creator>
<dc:creator>Komatsu, S.</dc:creator>
<dc:creator>Jiang, W. I.</dc:creator>
<dc:creator>Latiff, A. Q.</dc:creator>
<dc:creator>Patil, R. R.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:creator>Ashrafi, K.</dc:creator>
<dc:date>2025-06-29</dc:date>
<dc:identifier>doi:10.1101/2025.06.25.661551</dc:identifier>
<dc:title><![CDATA[Dietary restriction promotes neuronal resilience via ADIOL]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.26.661797v1?rss=1">
<title>
<![CDATA[
Photothrombotic Ischemic Thalamic Stroke in Mice Recapitulates Spontaneous Pain Features of Central Post-Stroke Pain in Humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.26.661797v1?rss=1</link>
<description><![CDATA[
Central post-stroke pain (CPSP) is a highly distressing condition that develops in 50% of people who suffer a thalamic stroke, and is typically unresponsive to current clinical treatments. Hypoxic damage to the ventral posterolateral (VPL) and ventral posteromedial (VPM) sensory thalamic nuclei, in particular, precipitates CPSP. One barrier to developing treatments for CPSP is the lack of preclinical models of thalamic ischemic stroke. In this study, we present a novel mouse model of CPSP induced through targeted photothrombotic ischemia. After eliciting hypoxia in the sensory thalamus of male mice, we assessed pain behaviors over a four-week period. Stroke-affected mice exhibited a persistent spontaneous facial grimace from day four to week four post-stroke, indicative of pain. Hind-paw mechanical hypersensitivity indicative of altered nociception, characteristic of VPL and VPM hemorrhagic CPSP models, was not detected in our model. Immunofluorescence analysis revealed increased activated microglia (Iba1) and reactive astrocytes (GFAP). Iba1 fluorescence intensity in the VPL thalamus-but not the VPM thalamus-correlated with the severity of facial grimace at four weeks post-stroke. Clustering based on behavioral phenotypes identified a subpopulation of mice in which grimace pain spontaneously resolved, by four weeks post-stroke, relative to sham controls, suggesting that this model can be used to understand how stroke recovery may influence pain chronification. This model provides a valuable tool to investigate the cellular and circuit mechanisms underlying CPSP after an ischemic thalamic stroke.

Significance StatementResearch into central post-stroke pain (CPSP) is hindered by the lack of preclinical models that match the clinical presentation of the condition. Although 87% of clinical strokes are ischemic, current CPSP preclinical models induce hemorrhagic strokes. To our knowledge, our mouse model is the first to induce CPSP by photothrombosis in the ventral posterolateral and posteromedial thalamus. Moreover, our study quantifies spontaneous pain, using a facial grimace score, in addition to nociception, whereas previous CPSP studies have relied exclusively on nociceptive tests. Finally, our model recapitulates some of the variability observed in clinical CPSP. We anticipate that this model will help dissect the roots of chronic pain after a thalamic stroke, and facilitate development of novel therapies for CPSP.
]]></description>
<dc:creator>Ford, J. B.</dc:creator>
<dc:creator>Tambou, O. N.</dc:creator>
<dc:creator>Weinrich, J. A.</dc:creator>
<dc:creator>Chatterjee, D.</dc:creator>
<dc:creator>Basbaum, A.</dc:creator>
<dc:creator>Paz, J. T.</dc:creator>
<dc:date>2025-06-29</dc:date>
<dc:identifier>doi:10.1101/2025.06.26.661797</dc:identifier>
<dc:title><![CDATA[Photothrombotic Ischemic Thalamic Stroke in Mice Recapitulates Spontaneous Pain Features of Central Post-Stroke Pain in Humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.26.661849v1?rss=1">
<title>
<![CDATA[
Simultaneous epigenomic profiling and regulatory activity measurement using e2MPRA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.26.661849v1?rss=1</link>
<description><![CDATA[
Cis-regulatory elements (CREs) have a major effect on phenotypes including disease. They are identified in a genome-wide manner by analyzing the binding of transcription factors (TFs), various co-factors and histone modifications in DNA using assays such as ChIP-seq, Cut&Tag and ATAC-seq. However, these assays are descriptive and require high-throughput technologies, such as massively parallel reporter assays (MPRAs), to test the functional activity and variant effect on these sequences. Currently, technologies that can simultaneously analyze both the regulatory function of a specific sequence and the TFs, cofactors and epigenomic modifications that determine it do not exist. Here, we developed enrichment followed by epigenomic profiling MPRA (e2MPRA), a novel technology that utilizes lentivirus-based MPRA to enrich for the integration of specific CREs into the genome followed by Cut&Tag or ATAC-seq targeted specifically for these sequences. This method allows to simultaneously analyze in a high-throughput manner regulatory activity, protein binding and epigenetic modification of thousands of candidate CREs and their variants. We demonstrate that e2MPRA can be used to dissect the epigenetic functions of TF motifs arranged in synthetic enhancers, as well as to analyze the effect of enhancer sequence variants on epigenetic modifications. In summary, this technology will increase our understanding of the regulatory code, its effect on the epigenome and how its alteration can lead to a variety of phenotypes including human disease.
]]></description>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Georgakopoulos Soares, I.</dc:creator>
<dc:creator>Bourque, G.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Inoue, F.</dc:creator>
<dc:date>2025-06-30</dc:date>
<dc:identifier>doi:10.1101/2025.06.26.661849</dc:identifier>
<dc:title><![CDATA[Simultaneous epigenomic profiling and regulatory activity measurement using e2MPRA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.26.661858v1?rss=1">
<title>
<![CDATA[
Luminal Vascular Dysfunction Drives Rapid Blood Brain Barrier Injury in Hyperglycemic Stroke: Key Roles for Luminal Glycocalyx and Complement 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.26.661858v1?rss=1</link>
<description><![CDATA[
BackgroundAcute hyperglycemia affects approximately 40% of stroke patients and is associated with worse outcomes. The underlying mechanisms linking this metabolic stress to stroke-induced brain injury remains unclear, and effective therapies are lacking.

MethodsIn a mouse model of acute hyperglycemic stroke, luminal disruption, blood-brain barrier (BBB) leakage, neurological deficit, motor function, and mortality were evaluated. Vascular luminal glycocalyx and complement activation were assessed by immunostaining, with glycocalyx loss confirmed by electron microscopy. Complement C3s causal role was tested using C3 knockout mice and site-targeted inhibition with CR2-Crry. To enhance translational relevance, post-mortem human stroke and control brains were immunostained to assess the association between endothelial glycocalyx loss and vascular complement activation. In a separate stroke patient cohort, soluble complement activation products were measured in pre-thrombectomy plasma, and their predictive value for modified Rankin Scale (mRS) outcomes evaluated using elastic net regression.

ResultsHyperglycemic stroke mice exhibited accelerated and more severe BBB breakdown, greater functional deficits, and higher mortality than normoglycemic controls, mirroring clinical observations. Acute hyperglycemia triggered rapid vascular luminal injury characterized by loss of endothelial luminal glycocalyx, luminal IgM/IgG deposition, and vascular complement C3 activation, leading to BBB disruption. This vascular luminal injury was corroborated in human stroke brain tissue. These luminal changes persisted despite glucose normalization and were exacerbated by reperfusion, driving injury into the brain parenchyma. Genetic and pharmacological approaches confirmed vascular complement activation as a causal driver of severe BBB disruption and poor outcomes. Importantly, site-targeted pharmacological inhibition of complement after reperfusion preserved BBB integrity and improved outcomes, defining a time-specific, luminal-directed strategy as a promising adjunct to thrombectomy. Notably, soluble complement activation markers in pre-thrombectomy stroke plasma predicted clinical outcomes, highlighting their potential as pre-intervention markers for patient stratification and tailored therapy.

ConclusionThis study reframes acute hyperglycemic stroke as a vascular luminal disorder, establishing a novel Metabolic-Complement-Vascular (MCV) axis linking metabolic stress to endothelial luminal glycocalyx loss, vascular complement activation, and BBB breakdown in both mice and humans. This new mechanistic understanding transforms the therapeutic landscape of hyperglycemic stroke, offering a potential time-defined, luminal-focused adjunct therapy alongside thrombectomy.

Clinical PerspectiveO_ST_ABSWhat Is New?C_ST_ABS- This study reframes hyperglycemic stroke as an acute vascular luminal problem, marked by rapid loss of endothelial luminal glycocalyx and complement C3 activation at the vascular luminal surface.
- The rapid luminal changes, identified in both rodent and human stroke brain tissues, establish a novel Metabolic-Complement-Vascular (MCV) axis linking metabolic stress to luminal damage, blood-brain barrier (BBB) breakdown, and injury progression into the brain parenchyma.
- The first clinical evidence that pre-thrombectomy plasma complement activation markers independently predict stroke outcomes -- laying the foundation for risk stratification before reperfusion and precision adjunct therapies.


What Are the Clinical Implications?- The rapidity and persistence of the MCV axis activation--even after glucose normalization--help explain the limited efficacy of insulin therapy, shifting the therapeutic focus from glycemic control to luminal-targeted interventions.
- Identification of a narrow but actionable window following reperfusion where complement C3 inhibition preserves the BBB and limits injury progression to the parenchyma, offers a promising adjunct to thrombectomy in metabolically vulnerable stroke patients.
- Targeting the vascular luminal surface reshapes the therapeutic landscape for hyperglycemic stroke by enabling systemic interventions--bypassing the challenge of BBB penetration, supporting rapid clinical translation using existing FDA-approved C3 inhibitors, and framing the endothelial glycocalyx as a promising area for therapeutic and diagnostic exploration in hyperglycemic stroke and broader cerebrovascular disease.
]]></description>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Frank, J. A.</dc:creator>
<dc:creator>Tan, C.</dc:creator>
<dc:creator>Lee, A. G.</dc:creator>
<dc:creator>Kopchock, R.</dc:creator>
<dc:creator>Chiang, T.</dc:creator>
<dc:creator>Kim, A.</dc:creator>
<dc:creator>Galvan, M.</dc:creator>
<dc:creator>Fraser, J. F.</dc:creator>
<dc:creator>Dornbos, D.</dc:creator>
<dc:creator>Aboul-Nour, H.</dc:creator>
<dc:creator>Millson, N.</dc:creator>
<dc:creator>Tomlinson, S.</dc:creator>
<dc:creator>McCullough, L. D.</dc:creator>
<dc:creator>Pennypacker, K.</dc:creator>
<dc:creator>Cheng, M. Y.</dc:creator>
<dc:creator>Bliss, T.</dc:creator>
<dc:creator>Steinberg, G. K.</dc:creator>
<dc:date>2025-06-30</dc:date>
<dc:identifier>doi:10.1101/2025.06.26.661858</dc:identifier>
<dc:title><![CDATA[Luminal Vascular Dysfunction Drives Rapid Blood Brain Barrier Injury in Hyperglycemic Stroke: Key Roles for Luminal Glycocalyx and Complement]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.26.661733v1?rss=1">
<title>
<![CDATA[
The gut microbiome shapes pharmacology and treatment outcomes for a key anti-inflammatory therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.26.661733v1?rss=1</link>
<description><![CDATA[
The human gut microbiome encodes a formidable metabolic repertoire that harvests nutrients from the diet, but these same pathways may also metabolize medications. Indeed, large screens have revealed extensive microbial metabolism of drugs in vitro, but the pharmacologic and clinical repercussions of microbiota-mediated metabolism in vivo remain to be discerned. As a proof-of-concept, we investigate how human gut microbes contribute to in vivo pharmacology and efficacy of a key anti-inflammatory drug, methotrexate (MTX). Specifically, we demonstrate that the gut microbiome shapes drug pharmacology in vivo in mice, both by directly metabolizing the drug and by inducing host pathways that promote drug metabolism. Moreover, interindividual variation in the human gut microbiome contributes to variation in pharmacokinetic (PK) profiles. When we quantified metabolites produced by microbes, we unexpectedly identified novel MTX metabolites, one of which, p-methylaminobenzoyl-L-glutamic acid (pMABG), was a major byproduct of microbial metabolism both in vitro and in vivo. Further, we find that a large proportion of patient-associated microbes are capable of metabolizing MTX. Finally, we show that microbial metabolism of MTX is linked to PK profiles and disease outcomes in a mouse model of inflammatory arthritis. Taken together, these findings provide evidence that the human gut microbiome causally contributes to drug pharmacology in vivo for a key anti-inflammatory drug through known and novel mechanisms. Our studies provide a framework for elucidating the clinical relevance of drug microbial metabolism in the context of treatment response. These results are a first step towards understanding and manipulating the human gut microbiome in the treatment of autoimmunity and the advancement of precision medicine for millions of patients taking MTX for immune or inflammatory conditions.

HighlightsO_LIThe gut microbiome impacts methotrexate (MTX) pharmacology in mice
C_LIO_LIThe human gut microbiome contributes to interindividual variation in MTX pharmacology
C_LIO_LIHuman gut microbes produce novel MTX metabolites, pMABG and 6-MPDA
C_LIO_LIMicrobial metabolism of MTX is linked to treatment outcomes
C_LI
]]></description>
<dc:creator>Villa Soto, V. S.</dc:creator>
<dc:creator>Degraeve, A. L.</dc:creator>
<dc:creator>Heath, C. M.</dc:creator>
<dc:creator>Orellana, D. A.</dc:creator>
<dc:creator>Reilly, E. R.</dc:creator>
<dc:creator>Mukherjee, M.</dc:creator>
<dc:creator>Brockert, J. G.</dc:creator>
<dc:creator>Dumlao, D. S.</dc:creator>
<dc:creator>Blank, R. B.</dc:creator>
<dc:creator>Perlmutter, N.</dc:creator>
<dc:creator>Yu, S.</dc:creator>
<dc:creator>Ashouri, J.</dc:creator>
<dc:creator>Scher, J. U.</dc:creator>
<dc:creator>Patterson, A. D.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:creator>Nayak, R. R.</dc:creator>
<dc:creator>Mukherjee, M. F.</dc:creator>
<dc:date>2025-07-01</dc:date>
<dc:identifier>doi:10.1101/2025.06.26.661733</dc:identifier>
<dc:title><![CDATA[The gut microbiome shapes pharmacology and treatment outcomes for a key anti-inflammatory therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.29.662044v1?rss=1">
<title>
<![CDATA[
A mitotic bookmark coordinates transcription and replication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.29.662044v1?rss=1</link>
<description><![CDATA[
Collisions between advancing replication forks and elongating transcripts pose a universal threat. During the rapid nuclear division cycles in early Drosophila embryos, coordinating transcription and replication is critical to reduce the risk of collisions. In each cycle, replication begins immediately after mitosis, while transcription starts 3 minutes later, overlapping with replication for the remainder of interphase. We previously showed that transcription depends on the coactivator Brd4, which forms hubs at active genes. Here, we show that Brd4 persists on mitotic chromosomes as bookmarks of transcriptional activity and, upon anaphase entry, recruits the replication activator Cdc7 to specify early-replicating genomic regions in the following interphase. Additionally, Cdc7 activity removes Brd4 bookmarks such that post-mitotic transcription occurs only after a new round of Brd4 hub assembly. Early initiation of replication while deferring initiation of transcription is proposed to allow unimpeded transcriptional elongation behind advancing replication forks. Supporting this, inhibiting Cdc7 delayed replication, stabilized Brd4 bookmarks, and resulted in premature transcription with elongation defects. We propose that Cdc7 triggers a functional switch in Brd4 that enforces temporal ordering of the initiation of transcription and replication, thereby minimizing collisions. This switching process might underlie the widespread correlation between transcriptional activity and early replication.
]]></description>
<dc:creator>Cho, C.-Y.</dc:creator>
<dc:creator>O'Farrell, P. H.</dc:creator>
<dc:date>2025-07-02</dc:date>
<dc:identifier>doi:10.1101/2025.06.29.662044</dc:identifier>
<dc:title><![CDATA[A mitotic bookmark coordinates transcription and replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.02.662771v1?rss=1">
<title>
<![CDATA[
Two parallel lineage-committed progenitors contribute to the developing brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.02.662771v1?rss=1</link>
<description><![CDATA[
The hindbrain is a life-sustaining brain region. In one model, a common neural progenitor generates all brain regions. Here our studies of mouse embryos and human pluripotent stem cells (hPSCs) support a different model: two parallel brain progenitors emerge simultaneously during gastrulation, anterior neural ectoderm (forebrain/midbrain progenitor) and posterior neural ectoderm (hindbrain progenitor). Not only are they lineage-committed to respectively form forebrain/midbrain vs. hindbrain in vitro, but they also have diverging chromatin landscapes foreshadowing future forebrain/midbrain vs. hindbrain identities. Leveraging these differences, we differentiated hPSCs into hindbrain rhombomere 5/6-specific motor neurons, hitherto difficult to generate in vitro. We postulate the brain is a composite organ emanating from two lineage-restricted progenitors; these dual progenitors may be evolutionarily conserved across 550 million years from hemichordates to mammals.
]]></description>
<dc:creator>Dundes, C. E.</dc:creator>
<dc:creator>Jokhai, R. T.</dc:creator>
<dc:creator>Ahsan, H.</dc:creator>
<dc:creator>Kang, R. S.</dc:creator>
<dc:creator>Salomon-Shulman, R. E. A.</dc:creator>
<dc:creator>Rajan, A.</dc:creator>
<dc:creator>Kim, Y. S.</dc:creator>
<dc:creator>Stanton, L. J.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Do, S.</dc:creator>
<dc:creator>McDonald, B. D.</dc:creator>
<dc:creator>Lopez, J. M. A.</dc:creator>
<dc:creator>Urrutia, H. A.</dc:creator>
<dc:creator>Greenfeld, H.</dc:creator>
<dc:creator>Wong, A.</dc:creator>
<dc:creator>Qu, Y.</dc:creator>
<dc:creator>Petkovic, A. S.</dc:creator>
<dc:creator>Miao, Y.</dc:creator>
<dc:creator>Garcia, K. C.</dc:creator>
<dc:creator>Monje, M.</dc:creator>
<dc:creator>Wagner, D. E.</dc:creator>
<dc:creator>Bronner, M. E.</dc:creator>
<dc:creator>Lowe, C. J.</dc:creator>
<dc:creator>Loh, K. M.</dc:creator>
<dc:date>2025-07-03</dc:date>
<dc:identifier>doi:10.1101/2025.07.02.662771</dc:identifier>
<dc:title><![CDATA[Two parallel lineage-committed progenitors contribute to the developing brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.30.662454v1?rss=1">
<title>
<![CDATA[
Multimodal sensory overload in dopamine-deficient larval zebrafish leads to paradoxical kinesia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.30.662454v1?rss=1</link>
<description><![CDATA[
Paradoxical kinesia--the temporary alleviation of motor deficits by powerful, urgent stimuli in Parkinsons disease (PD)--remains poorly understood at the neural circuit level. Through chemo-genetic ablation of tyrosine hydroxylase-expressing neurons in larval zebrafish and brain-wide calcium imaging under head-fixed, tail-free conditions, we uncovered a neural mechanism underlying this phenomenon. While catecholamine (CA)-deficient larvae exhibited severe locomotor deficits during free swimming, they showed paradoxical recovery of tail movements during whole-brain neural activity imaging. This locomotor recovery was accompanied by a significantly increased number of active neurons in the midbrain and hindbrain, but with reduced firing rates. Further analyses across 2158 anatomically defined regions allowed us to uncover a subset of regions, genes, and neurotransmitter types. GABAergic neurons were found to primarily account for the hyperactivity in the hindbrain, while glutamatergic neurons accounted for the hyperactivity in the midbrain. Hierarchical clustering of neuronal activity with tail movements revealed distinct motor- and non-motor-associated hyperactive clusters in the hindbrain and midbrain, respectively. We identified the Mesencephalic Locomotor Region (MLR) sandwiched between these domains, with enhanced glutamatergic firing rate and cholinergic activation. Furthermore, we found that Telencephalic corticotropin-releasing factor b (crhb) expressing neurons play a crucial role in mediating stress-response to the tectum, which in turn triggers a cascade of neuronal hyperactivity downstream via MLR. These findings reveal a neural mechanism that links stress-induced sensory processing with motor control systems in the absence of regulatory feedback from catecholaminergic neurons, suggesting a direct, unmodulated pathway that bypasses typical inhibitory controls.
]]></description>
<dc:creator>Bose, K.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:creator>Bhardwaj, K.</dc:creator>
<dc:date>2025-07-04</dc:date>
<dc:identifier>doi:10.1101/2025.06.30.662454</dc:identifier>
<dc:title><![CDATA[Multimodal sensory overload in dopamine-deficient larval zebrafish leads to paradoxical kinesia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.01.661217v1?rss=1">
<title>
<![CDATA[
Differential Assembly of Mouse and Human Tumor Microenvironments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.01.661217v1?rss=1</link>
<description><![CDATA[
Mouse models are frequently used to develop treatments for human cancer. Yet, we lack a comprehensive understanding of the comparative organization of mouse and human tumor microenvironments (mu/huTMEs). Through immunoprofiling of commonly used mouse models, we found that the immune composition of most muTMEs resemble poorly infiltrated human tumors extensively biased toward high macrophages densities. Relatedly, we discover species-specific biases of chemokine expression networks, factors which drive TMEs assembly. Further, assessing coarse cellular networks, we find conserved correlations between some immune cell frequencies, while other relationships only appear conserved in the huTMEs matching muTME profiles. Despite this variable alignment, we define robust cell type-specific gene expression programs conserved in TMEs across species and cohorts and identify ones that are coordinated between cell populations in both species. Together, we isolate and offer methods to study the multiple areas of hazard and opportunities for using mice to model human cancer.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=192 HEIGHT=200 SRC="FIGDIR/small/661217v1_ufig1.gif" ALT="Figure 1">
View larger version (71K):
org.highwire.dtl.DTLVardef@22bcf7org.highwire.dtl.DTLVardef@a410b7org.highwire.dtl.DTLVardef@1455eb5org.highwire.dtl.DTLVardef@14cc6f5_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Courau, T.</dc:creator>
<dc:creator>Jaszczak, R. G.</dc:creator>
<dc:creator>Samad, B.</dc:creator>
<dc:creator>Flynn, E.</dc:creator>
<dc:creator>Reeder, G. C.</dc:creator>
<dc:creator>Chew, N. W.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Ray, A.</dc:creator>
<dc:creator>Wismer, H.</dc:creator>
<dc:creator>Bunis, D.</dc:creator>
<dc:creator>Lupin-Jimenez, L.</dc:creator>
<dc:creator>Gavil, N. V.</dc:creator>
<dc:creator>Masopust, D.</dc:creator>
<dc:creator>Graham, J. P.</dc:creator>
<dc:creator>Skelly, D. A.</dc:creator>
<dc:creator>Vesco, X.</dc:creator>
<dc:creator>Liu, E. T.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2025-07-05</dc:date>
<dc:identifier>doi:10.1101/2025.07.01.661217</dc:identifier>
<dc:title><![CDATA[Differential Assembly of Mouse and Human Tumor Microenvironments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.01.662602v1?rss=1">
<title>
<![CDATA[
Labile iron pool dynamics do not drive ferroptosis potentiation in colorectal cancer cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.01.662602v1?rss=1</link>
<description><![CDATA[
Colorectal cancer (CRC) is the second leading cause of cancer-related mortality in the United States. CRC tumors exhibit aberrant iron accumulation, which supports tumor cell proliferation through multiple metabolic pathways. However, the oncogenic benefits of elevated iron must be counterbalanced by its potential to catalyze oxidative damage via reactive oxygen species generated from labile, redox-active iron. Ferroptosis is a regulated, non-apoptotic form of cell death characterized by iron-dependent lipid peroxidation. This process is tightly controlled by the selenoenzyme glutathione peroxidase 4 (GPX4), which reduces lipid peroxides and can be pharmacologically inhibited by agents such as RSL3 and JKE1674. A key source of redox-active iron is the labile iron pool (LIP), yet its role in regulating ferroptosis remains incompletely defined. To examine this, we supplemented CRC cells with exogenous iron following pharmacologic induction of ferroptosis. Iron supplementation significantly reduced cell viability, suggesting that expansion of the LIP potentiates ferroptotic cell death. However, whether ferroptosis is accompanied by dynamic changes in the LIP, and if such changes are mechanistically required for its potentiation, were unknown. To further characterize this response, we profiled the expression of iron regulatory genes under ferroptotic conditions and observed no change in transcriptional response in iron homeostasis genes. Using a reactivity-based probe of labile iron, we found that the LIP did not measurably increase during ferroptosis induction with GPX4 inhibition or inhibition of the SLC7A11 cysteine/glutamate antiporter. These findings suggest that the LIP does not expand upon pharmacological initiated ferroptosis, despite the potentiating effect of exogenous iron supplementation.
]]></description>
<dc:creator>Ponnusamy, V.</dc:creator>
<dc:creator>Randall, D. R.</dc:creator>
<dc:creator>Lee, Z.</dc:creator>
<dc:creator>Das, N. K.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:creator>Buscher, K.</dc:creator>
<dc:creator>Solanki, S.</dc:creator>
<dc:creator>Renslo, A.</dc:creator>
<dc:creator>Shah, Y. M.</dc:creator>
<dc:creator>Hsu, P. P.</dc:creator>
<dc:date>2025-07-05</dc:date>
<dc:identifier>doi:10.1101/2025.07.01.662602</dc:identifier>
<dc:title><![CDATA[Labile iron pool dynamics do not drive ferroptosis potentiation in colorectal cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.01.662675v1?rss=1">
<title>
<![CDATA[
TOMM40 Knockdown in Macrophages Inhibits Oxidized LDL-induced NLRP3 Activation and Promotes LXR-β Mediated Cholesterol Transporter Gene Expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.01.662675v1?rss=1</link>
<description><![CDATA[
Atherogenesis has been shown in mice to be dependent on activation of the NLRP3 inflammasome, a cytosolic innate immune sensor activated by a broad range of pathogen and damage associated molecular patterns, including oxidized LDL (oxLDL) in atherosclerosis. Previous work from our group has shown that knockdown of Translocase of Outer Mitochondrial Membrane 40 (TOMM40), which encodes a component of the mitochondrial importer TOM complex, increases expression and activity of the nuclear receptor-family transcription factor, liver X receptor (LXR) in hepatocytes. As LXR agonists have been shown to inhibit NLRP3 activation, we confirmed the prediction that TOMM40 knockdown has this effect in THP-1 monocyte-derived macrophages. Further, we demonstrated that TOMM40 KD upregulates LXR-mediated macrophage expression of the ABCA1 and ABCG1 genes which encode transporters that promote cellular cholesterol efflux, the first step in reverse cholesterol transport to the liver. Taken together, these findings identify a novel mechanism whereby increased LXR activity induced by suppression of TOMM40 expression in macrophages may retard atherogenesis both by inhibiting inflammation and promoting reverse cholesterol transport.
]]></description>
<dc:creator>Chelluri, S. S.</dc:creator>
<dc:creator>Yang, N. V.</dc:creator>
<dc:creator>Theusch, E.</dc:creator>
<dc:creator>Krauss, R. M.</dc:creator>
<dc:date>2025-07-05</dc:date>
<dc:identifier>doi:10.1101/2025.07.01.662675</dc:identifier>
<dc:title><![CDATA[TOMM40 Knockdown in Macrophages Inhibits Oxidized LDL-induced NLRP3 Activation and Promotes LXR-β Mediated Cholesterol Transporter Gene Expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.04.663249v1?rss=1">
<title>
<![CDATA[
Electrocorticographic Detection of Speech Networks in Glioma-infiltrated Cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.04.663249v1?rss=1</link>
<description><![CDATA[
Direct cortical stimulation (DCS) is the clinical gold standard for identifying functional cortex in the human brain, which is essential for the safe removal of brain lesions. Defining the electro-physiological properties of DCS- positive cortical regions may facilitate the identification of critical language regions, thereby permitting safe glioma resections in communities without access. Leveraging a multicenter electrophysiologic dataset of DCS- positive language regions spatially matched with subdural arrays, we analyzed regions identified as functionally critical (DCS+) versus functionally non-critical (DCS-) during intraoperative language mapping. In IDH-mutant gliomas, DCS+ regions exhibited significantly greater speech-related neural activity and enhanced encoding and decoding of linguistic and semantic features. We demonstrate that resting-state classifiers distinguish DCS+ from DCS- regions in IDH-mutant tumors. Task-based and resting-state electrophysiologic distinctions were pathology-specific and not present in IDH-wildtype glioblastomas. These findings may accelerate DCS mapping by guiding surgeons to priority regions, improving efficiency, and patient outcomes.
]]></description>
<dc:creator>Ambati, V. S.</dc:creator>
<dc:creator>Persad, A. S.</dc:creator>
<dc:creator>Kaur, J.</dc:creator>
<dc:creator>Herr, S.</dc:creator>
<dc:creator>Cunningham, E.</dc:creator>
<dc:creator>Dada, A.</dc:creator>
<dc:creator>Ekert, J. O.</dc:creator>
<dc:creator>Greicius, Q.</dc:creator>
<dc:creator>Silva, A.</dc:creator>
<dc:creator>McMillan Villalobos, P.</dc:creator>
<dc:creator>Olshausen, N.</dc:creator>
<dc:creator>Sibih, Y.</dc:creator>
<dc:creator>Oten, S.</dc:creator>
<dc:creator>Yamada, H. S.</dc:creator>
<dc:creator>Umbach, G.</dc:creator>
<dc:creator>Aabedi, A. A.</dc:creator>
<dc:creator>Al-Holou, W.</dc:creator>
<dc:creator>Young, J.</dc:creator>
<dc:creator>Sushil, M.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Berger, M. S.</dc:creator>
<dc:creator>Brang, D.</dc:creator>
<dc:creator>Hervey-Jumper, S. L.</dc:creator>
<dc:date>2025-07-05</dc:date>
<dc:identifier>doi:10.1101/2025.07.04.663249</dc:identifier>
<dc:title><![CDATA[Electrocorticographic Detection of Speech Networks in Glioma-infiltrated Cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.04.663231v1?rss=1">
<title>
<![CDATA[
Product-stabilized filamentation by human glutamine synthetase allosterically tunes metabolic activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.04.663231v1?rss=1</link>
<description><![CDATA[
To maintain metabolic homeostasis, enzymes must adapt to fluctuating nutrient levels through mechanisms beyond gene expression. Here, we demonstrate that human glutamine synthetase (GS) can reversibly polymerize into filaments aided by a composite binding site formed at the filament interface by the product, glutamine. Time-resolved cryo-electron microscopy (cryo-EM) confirms that glutamine binding stabilizes these filaments, which in turn exhibit reduced catalytic specificity for ammonia at physiological concentrations. This inhibition appears induced by a conformational change that remodulates the active site loop ensemble gating substrate entry. Metadynamics ensemble refinement revealed >10 [A] conformational range for the active site loop and that the loop is stabilized by transient contacts. This disorder is significant, as we show that the transient contacts which stabilize this loop in a closed conformation are essential for catalysis both in vitro and in cells. We propose that GS filament formation constitutes a negative-feedback mechanism, directly linking product concentration to the structural and functional remodeling of the enzyme.
]]></description>
<dc:creator>Greene, E.</dc:creator>
<dc:creator>Muniz, R.</dc:creator>
<dc:creator>Yamamura, H.</dc:creator>
<dc:creator>Hoff, S. E.</dc:creator>
<dc:creator>Bajaj, P.</dc:creator>
<dc:creator>Lee, D. J.</dc:creator>
<dc:creator>Thompson, E. M.</dc:creator>
<dc:creator>Arada, A.</dc:creator>
<dc:creator>Lee, G. M.</dc:creator>
<dc:creator>Bonomi, M.</dc:creator>
<dc:creator>Kollman, J. M.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2025-07-06</dc:date>
<dc:identifier>doi:10.1101/2025.07.04.663231</dc:identifier>
<dc:title><![CDATA[Product-stabilized filamentation by human glutamine synthetase allosterically tunes metabolic activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.04.663190v1?rss=1">
<title>
<![CDATA[
Blocking Osteoprotegerin Reprograms Cancer Associated Fibroblast to Promotes Immune Infiltration into the Tumor Microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.04.663190v1?rss=1</link>
<description><![CDATA[
AbstractThe stromal compartment of many solid tumors plays a critical role in shaping an immunosuppressive microenvironment that limits the effectiveness of immune-based therapies1. Among stromal constituents, cancer-associated fibroblasts (CAFs) have emerged as key regulators of antitumor immunity2-5. Here, we identify a distinct subset of CAFs in both murine and human stroma-rich cancers that secrete osteoprotegerin (OPG)-- a soluble decoy receptor that neutralizes receptor activator of nuclear factor kappa-B ligand (RANKL) and TNF-related apoptosis-inducing ligand (TRAIL), both of which are involved in T cell function. In vitro, OPG directly impairs CD8 T cell-mediated killing of target cells. In murine models of pancreatic and breast cancer, antibody-mediated blockade of OPG promotes robust immune infiltration into the tumor microenvironment, leading to significant tumor regression. Stromal profiling revealed that OPG blockade induces a shift in CAF cells--reducing immunosuppressive OPG fibroblasts while expanding interferon-responsive fibroblasts, thus recalibrating the tumor stroma toward a pro-immunogenic landscape. These findings uncover a previously unrecognized mechanism of stromal immune suppression and highlight OPG as a stromal immune checkpoint controlling CD8 T cell infiltration. Targeting OPG may offer a novel therapeutic strategy to convert immunologically "cold" tumors into T cell-infiltrated, tumor microenvironment.
]]></description>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Apostolopoulou, H.</dc:creator>
<dc:creator>Sun, I. H.</dc:creator>
<dc:creator>Bains, A.</dc:creator>
<dc:creator>Gibbs, D.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Alliston, T.</dc:creator>
<dc:creator>Maker, A.</dc:creator>
<dc:creator>Tlsty, T.</dc:creator>
<dc:creator>Ntranos, V.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Bhushan, A.</dc:creator>
<dc:date>2025-07-07</dc:date>
<dc:identifier>doi:10.1101/2025.07.04.663190</dc:identifier>
<dc:title><![CDATA[Blocking Osteoprotegerin Reprograms Cancer Associated Fibroblast to Promotes Immune Infiltration into the Tumor Microenvironment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.07.663234v1?rss=1">
<title>
<![CDATA[
Beating the gold standard: A review of Mycobacterium tuberculosis lysis using bead beating and the need for standardization. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.07.663234v1?rss=1</link>
<description><![CDATA[
Bead beating is widely used for mechanical lysis of Mycobacterium tuberculosis, a bacterium with a highly resistant, lipid-rich cell wall. Despite its status as a de facto gold standard for mycobacterial lysis, there is no standardized protocol for bead beating, resulting in significant variability across studies. We conducted a literature review of 73 studies, identifying 38 with explicit mycobacterial bead beating protocols. Our analysis revealed heterogeneity in bead types, sizes, device models, and operational parameters, with 37% of studies failing to report critical details such as lysis speed. We experimentally assessed the impact of key variables--tube type, bead quantity, and device settings--on lysis efficiency using qPCR of M. tuberculosis DNA. Results showed that even minor changes, such as tube shape or bead volume, can significantly affect DNA yield. These findings underscore the need for standardized bead-beating protocols to improve reproducibility and comparability. Future efforts should prioritize developing consensus methods tailored to sample type and analytical application.
]]></description>
<dc:creator>Limberis, J. D.</dc:creator>
<dc:creator>Metcalfe, J. Z.</dc:creator>
<dc:date>2025-07-07</dc:date>
<dc:identifier>doi:10.1101/2025.07.07.663234</dc:identifier>
<dc:title><![CDATA[Beating the gold standard: A review of Mycobacterium tuberculosis lysis using bead beating and the need for standardization.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.07.661868v1?rss=1">
<title>
<![CDATA[
High-throughput multi-camera array microscope platform for automated 3D behavioral analysis of freely swimming zebrafish larvae 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.07.661868v1?rss=1</link>
<description><![CDATA[
Understanding the behavioral and morphological dynamics of moving model organisms like the zebrafish larvae requires accurate, high-throughput 3D analysis. However, traditional single-view 2D video tracking fails to capture the full scope of natural 3D movements and postural dynamics. Here, we present a novel high-throughput 24-camera array microscope with a co-designed "mirrored well plate" that allows for snapshot imaging of up to 48 wells over a 118 mm x 82 mm field of view from two orthogonal directions (i.e., a top-view and side-view). Accurate 3D position estimation and tracking is achieved with an efficient machine learning algorithm that scales well to high-throughput measurements. The proposed approach automates parallelized 3D model organism behavioral analysis, providing 3D skeletal tracking, swim bladder morphological dynamics, and kinematics of up to 48 swimming zebrafish larvae at up to several hundred frames per second. The result is an efficient and scalable solution for high-throughput 3D behavioral studies with broad compatibility with standard workflows across laboratories and procedures working with pharmacology, toxicology, and neuroscience.
]]></description>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Kreiss, L.</dc:creator>
<dc:creator>Reamey, P.</dc:creator>
<dc:creator>Pierce, L. X.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Da Luz, R.</dc:creator>
<dc:creator>Chaware, A.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Cook, C. B.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Lerner, J. F.</dc:creator>
<dc:creator>Doman, J.</dc:creator>
<dc:creator>Begue, A.</dc:creator>
<dc:creator>Efromson, J.</dc:creator>
<dc:creator>Harfouche, M.</dc:creator>
<dc:creator>Horstmeyer, G.</dc:creator>
<dc:creator>McCarroll, M. N.</dc:creator>
<dc:creator>Horstmeyer, R.</dc:creator>
<dc:date>2025-07-08</dc:date>
<dc:identifier>doi:10.1101/2025.07.07.661868</dc:identifier>
<dc:title><![CDATA[High-throughput multi-camera array microscope platform for automated 3D behavioral analysis of freely swimming zebrafish larvae]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.07.663529v1?rss=1">
<title>
<![CDATA[
Benchmarking Large Language Models for Predictive Modeling in Biomedical Research With a Focus on Reproductive Health 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.07.663529v1?rss=1</link>
<description><![CDATA[
Generative AI, particularly large language models (LLMs), is increasingly being used in computational biology to support code generation for data analysis. In this study, we evaluated the ability of LLMs to generate functional R and Python code for predictive modeling tasks, leveraging standardized molecular datasets from several recent DREAM (Dialogue for Reverse Engineering Assessments and Methods) Challenges focused on reproductive health. We assessed LLM performance across four predictive tasks derived from three DREAM challenges: gestational age regression from gene expression, gestational age regression from DNA methylation profiles, and classification of preterm birth and early preterm birth from microbiome data. LLMs were prompted with task descriptions, data locations, and target outcomes. LLM-generated code was then run to fit and apply prediction models and generate graphics, and they were ranked based on their success in completing the tasks and achieving strong test set performance. Among the eight LLMs tested, o3-mini-high, 4o, DeepseekR1 and Gemini 2.0 completed at least one task without error. Overall, R code generation was more successful (14/16 tasks) than Python (7/16), attributed to the utility of Bioconductor packages for querying Gene Expression Omnibus data. OpenAIs o3-mini-high outperformed others, completing 7/8 tasks. Test set performance of the top LLM matched or exceeded top-performing teams from the original DREAM challenges. These findings underscore the potential of LLMs to enhance exploratory analysis and democratize access to predictive modeling in omics by automating key components of analysis pipelines, and highlight the potential to increase research output when conducting analyses of standardized datasets from public repositories.
]]></description>
<dc:creator>Sarwal, R.</dc:creator>
<dc:creator>Tarca, V.</dc:creator>
<dc:creator>Kalavros, N.</dc:creator>
<dc:creator>Bhatti, G.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Butte, A. J.</dc:creator>
<dc:creator>Romero, R. J.</dc:creator>
<dc:creator>Stolovitzky, G.</dc:creator>
<dc:creator>Oskotsky, T. T.</dc:creator>
<dc:creator>Tarca, A. L.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2025-07-10</dc:date>
<dc:identifier>doi:10.1101/2025.07.07.663529</dc:identifier>
<dc:title><![CDATA[Benchmarking Large Language Models for Predictive Modeling in Biomedical Research With a Focus on Reproductive Health]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.08.663653v1?rss=1">
<title>
<![CDATA[
Clinical and molecular characterisation of primary refractoriness to atezolizumab plus bevacizumab in patients with unresectable hepatocellular carcinoma. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.08.663653v1?rss=1</link>
<description><![CDATA[
BackgroundDespite improved outcomes with atezolizumab plus bevacizumab (A+B) in hepatocellular carcinoma (HCC), primary refractoriness (PRef), characterised by early progression or short-lived disease stabilisation following treatment, remains a significant and poorly understood clinical challenge.

MethodsWe analysed 1296 patients with HCC and Child-Pugh A liver cirrhosis treated with frontline A+B (AB-real) and validated findings in 645 trial participants recruited to IMbrave150 and GO30140. PRef was defined by Society for the Immunotherapy of Cancer (SITC) criteria as progressive disease in the first 6 months after treatment initiation. Patients who achieved complete response, partial response or stable disease for [&ge;] 6 months were classified as responders. We performed a multi-parametric analysis of pre-treatment tumour tissue including machine learning-based quantification of tumour-infiltrating lymphocytes, imaging mass cytometry and RNA sequencing (RNAseq) to evaluate differences in the tumour microenvironment (TME) of PRef versus responding patients. We employed conditional inference tree analyses to provide a hierarchical organisation of determinants of PRef.

ResultsAmong 677 AB-real and 378 trial patients evaluable by SITC criteria, PRef identified inferior median OS in comparison with responding patients (AB-real: 7.3 vs. 31.5 months, HR 3.7, 95%CI 2.8-8.5, p<0.001; Trials: 10.8 vs. NR, HR 4.6, 95%CI 3.3-6.3, p<0.001). PRef patients exhibited higher baseline systemic inflammation (neutrophil-to-lymphocyte ratio, NLR [&ge;]3), a distinctively immunosuppressive TME enriched in CD163+ tumour-associated macrophages and a higher Treg/Teff ratio. RNAseq of tumour tissue demonstrated lower intrinsic immunogenicity in PRef samples, characterised by repressed IFN-{gamma} and Teff signatures, with elevated myeloid infiltration. Conditional inference tree analysis identified IFN-{gamma} signature downregulation combined with NLR [&ge;]3 as the strongest contributor of PRef.

ConclusionsPRef to A+B identifies a distinct biological entity characterised by unopposed systemic inflammation, myeloid cell infiltration and T-cell depletion. Targeting myeloid-mediated immunosuppression, particularly in patients with low IFN-{gamma} signature expression and elevated NLR might enhance responsiveness to A+B.

HighlightsO_LIPrimary refractoriness to atezolizumab plus bevacizumab in hepatocellular carcinoma, as defined by SITC criteria, is associated with poor clinical outcomes.
C_LIO_LITumour microenvironment profiling reveals an immunosuppressive phenotype characterized by high myeloid infiltration, reduced interferon-{gamma} signalling, and T-cell depletion.
C_LIO_LIThe combination of systemic inflammation and low IFN-{gamma} signature expression strongly predicts primary refractoriness and may inform therapeutic decision-making.
C_LI
]]></description>
<dc:creator>Lombardi, P.</dc:creator>
<dc:creator>Ramon-Gil, E.</dc:creator>
<dc:creator>Brunetti, L.</dc:creator>
<dc:creator>Manfredi, G. F.</dc:creator>
<dc:creator>Merces, G.</dc:creator>
<dc:creator>Fulgenzi, C. A. M.</dc:creator>
<dc:creator>DAlessio, A.</dc:creator>
<dc:creator>Torkpour, A.</dc:creator>
<dc:creator>Celsa, C.</dc:creator>
<dc:creator>Stefanini, B.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Crowley, F.</dc:creator>
<dc:creator>Marron, T. U.</dc:creator>
<dc:creator>Saeed, A.</dc:creator>
<dc:creator>Pinter, M.</dc:creator>
<dc:creator>Scheiner, B.</dc:creator>
<dc:creator>Huang, Y.-H.</dc:creator>
<dc:creator>Lee, P.-C.</dc:creator>
<dc:creator>Nishida, N.</dc:creator>
<dc:creator>Po-Ting Lin, R.</dc:creator>
<dc:creator>Dalbeni, A.</dc:creator>
<dc:creator>Vivaldi, C.</dc:creator>
<dc:creator>Masi, G.</dc:creator>
<dc:creator>Rohlen, N.</dc:creator>
<dc:creator>von Felden, J.</dc:creator>
<dc:creator>Kaseb, A.</dc:creator>
<dc:creator>Galle, P. R.</dc:creator>
<dc:creator>Kudo, M.</dc:creator>
<dc:creator>Hsu, W.-F.</dc:creator>
<dc:creator>Rimassa, L.</dc:creator>
<dc:creator>Parisi, A.</dc:creator>
<dc:creator>Kelley, R. K.</dc:creator>
<dc:creator>Toyoda, H.</dc:creator>
<dc:creator>Pirisi, M.</dc:creator>
<dc:creator>Jabar, R. F.</dc:creator>
<dc:creator>Rakaee, M.</dc:creator>
<dc:creator>Cabibbo, G.</dc:creator>
<dc:creator>Camma, C.</dc:creator>
<dc:creator>Piscaglia, F.</dc:creator>
<dc:creator>Hwang, S.</dc:creator>
<dc:creator>Shin, D. J.</dc:creator>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Daniele, G.</dc:creator>
<dc:creator>Mann, D.</dc:creator>
<dc:creator>Chon, H. J.</dc:creator>
<dc:creator>Leslie, J.</dc:creator>
<dc:creator>Pin</dc:creator>
<dc:date>2025-07-10</dc:date>
<dc:identifier>doi:10.1101/2025.07.08.663653</dc:identifier>
<dc:title><![CDATA[Clinical and molecular characterisation of primary refractoriness to atezolizumab plus bevacizumab in patients with unresectable hepatocellular carcinoma.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.08.661805v1?rss=1">
<title>
<![CDATA[
Selective Sensitivity of Ph-like B-ALL to BRG1 Inhibition Reveals a Novel Targeted Therapy Strategy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.08.661805v1?rss=1</link>
<description><![CDATA[
Despite therapeutic advances, high-risk subtypes of B-cell acute lymphoblastic leukemia (B-ALL) such as Philadelphia chromosome-like (Ph-like) and KMT2A-rearranged (KMT2A-R) remain a formidable clinical challenge. BRG1 (gene name SMARCA4), the ATPase subunit of the SWI/SNF chromatin-remodeling complex, has been extensively studied in solid tumors, where inactivating mutations are linked to aggressive disease and poor prognosis. Although BRG1 is known to be essential for early B cell development, its role in B-ALL remains poorly understood. Therefore, we investigated the therapeutic relevance of BRG1 in high-risk B-ALL. Meta-analysis of gene expression data revealed that BRG1-inactivating mutations are exceedingly rare (0.35%) in B-ALL, suggesting that intact BRG1 function may be critical for leukemogenesis. Subtype-specific analyses revealed that elevated BRG1 expression is associated with significantly shorter overall survival in children with Ph-like B-ALL, while the opposite trend was observed in KMT2A-R B-ALL. We confirmed higher BRG1 expressions in Ph-like compared to KMT2A-R B-ALL via gene expression analysis, RT-PCR, and Western blotting. The pharmacologic inhibition of BRG1 using two selective inhibitors, BRM014 and FHD-286, revealed marked sensitivity in Ph-like B-ALL cell lines, whereas KMT2A-R B-ALL was resistant. Mechanistically, we found that BRG1 inhibition results in cell cycle arrest via downregulation of cell cycle regulators (CCND3, CDK4, CDK6, E2F1, and MYC) and upregulation of the cell cycle inhibitor CDKN1B (p27). Importantly, treatment with FHD-286 significantly prolonged the survival of NSG mice engrafted with Ph-like B-ALL cells. Taken together, these findings establish BRG1 as a critical, subtype-specific dependency in Ph-like B-ALL and demonstrate that its pharmacologic inhibition effectively suppresses leukemic cell proliferation through induction of cell cycle arrest. The pronounced in vitro sensitivity and improved in vivo survival upon BRG1 inhibition provide compelling preclinical evidence for its therapeutic targeting. These results support the advancement of BRG1-directed strategies as a viable treatment approach for patients with Ph-like B-ALL, with the potential to improve outcomes in this high-risk population.

HighlightsO_LIHigher levels of BRG1 correlate to poor clinical outcomes in Ph-like but KMT2A-R B-ALL
C_LIO_LIInhibition of BRG1 induces cell cycle arrest in Ph-like cells in vitro and extends the survival of mice in pre-clinical in vivo studies
C_LI

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/661805v1_ufig1.gif" ALT="Figure 1">
View larger version (56K):
org.highwire.dtl.DTLVardef@1b0a6cforg.highwire.dtl.DTLVardef@977257org.highwire.dtl.DTLVardef@d3bf7forg.highwire.dtl.DTLVardef@1c12d52_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Ayyadevara, V. S. S. A.</dc:creator>
<dc:creator>Gaur, S.</dc:creator>
<dc:creator>Paik, A.</dc:creator>
<dc:creator>Perencsik, R.</dc:creator>
<dc:creator>Toma, M. M.</dc:creator>
<dc:creator>Wertheim, G.</dc:creator>
<dc:creator>Geng, H.</dc:creator>
<dc:creator>Skorski, T.</dc:creator>
<dc:creator>Swaminathan, S.</dc:creator>
<dc:creator>Hurtz, C.</dc:creator>
<dc:date>2025-07-11</dc:date>
<dc:identifier>doi:10.1101/2025.07.08.661805</dc:identifier>
<dc:title><![CDATA[Selective Sensitivity of Ph-like B-ALL to BRG1 Inhibition Reveals a Novel Targeted Therapy Strategy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.08.663122v1?rss=1">
<title>
<![CDATA[
Cell atlas of the developing human meninges reveals a dura origin of meningioma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.08.663122v1?rss=1</link>
<description><![CDATA[
The vertebrate central nervous system is enveloped by the meninges, consisting of the pia, arachnoid, and dura layers. The arachnoid is hypothesised to give rise to the most common primary intracranial tumours, meningiomas. However, molecular evidence supporting this hypothesis is lacking. There are no effective medical therapies to treat meningiomas that are resistant to local interventions, encumbered by our limited understanding of their cellular origin. To address this limitation in our understanding of meningioma biology, we generated a comprehensive reference single cell and spatial transcriptomic atlas of human fetal meninges at post-conceptional weeks 5-13. We found that the meningeal layers develop concurrently, and identified an inner CDH1-positive dura cell layer expressing tight junction genes consistent with barrier function. We show that transcriptionally, meningioma cells resemble dura-lineage cells, and that common meningioma driver genes were expressed preferentially in the dura lineage. Our findings suggest that meningiomas originate from dura lineage cells.

HIGHLIGHTSO_LIscRNA-seq and spatial transcriptomics reveals architecture of human fetal meninges development
C_LIO_LIMeningeal layers are formed concurrently, by a gradual refinement of cell states
C_LIO_LIA CDH1+ inner dura sublayer expresses tight junction genes like the arachnoid barrier
C_LIO_LIMeningioma tumours likely originate from inner dura lineage cells, not arachnoid
C_LI
]]></description>
<dc:creator>Vinsland, E.</dc:creator>
<dc:creator>Marco Salas, S.</dc:creator>
<dc:creator>Kapustova, I.</dc:creator>
<dc:creator>Hu, L.</dc:creator>
<dc:creator>Webb, S.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>He, X.</dc:creator>
<dc:creator>Nilsson, M.</dc:creator>
<dc:creator>Hanifa, M.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Persson, O.</dc:creator>
<dc:creator>Raleigh, D. R.</dc:creator>
<dc:creator>Sundstrom, E.</dc:creator>
<dc:creator>Lonnerberg, P.</dc:creator>
<dc:creator>Linnarsson, S.</dc:creator>
<dc:date>2025-07-11</dc:date>
<dc:identifier>doi:10.1101/2025.07.08.663122</dc:identifier>
<dc:title><![CDATA[Cell atlas of the developing human meninges reveals a dura origin of meningioma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.08.663709v1?rss=1">
<title>
<![CDATA[
Tim-3 Promotes Early Differentiation of Tbet+ Effector T Cells During Acute Viral Infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.08.663709v1?rss=1</link>
<description><![CDATA[
The transmembrane protein Tim-3 has received significant attention in recent years as a possible immunotherapy target. This is due to its robust expression on dysfunctional, exhausted, T cells found in the settings of cancer and chronic infection and biochemical evidence suggesting an inhibitory function of Tim-3. However, numerous clinical trials of putative Tim-3 blocking antibodies have yielded modest benefits, at best, in clinical trials for various cancers. Thus, there is a need to more fully understand the function of Tim-3 in vivo. Here we have studied the function of Tim-3 early during a T cell response to LCMV Armstrong, which causes an acute viral infection in mice. We show that Tim-3 is rapidly expressed after infection and that the expression of Tim-3 is associated with acquisition of a type I effector phenotype, including expression of T-bet and downregulation of Tcf-1, by both CD4+ and CD8+ T cells. In addition, we demonstrate that knockout or cytoplasmic truncation of Tim-3 attenuates the acquisition of the effector program by T cells after LCMV Armstrong infection. Together, these data help to clarify the role of Tim-3 during acute infection.
]]></description>
<dc:creator>Manandhar, P.</dc:creator>
<dc:creator>Landy, E.</dc:creator>
<dc:creator>Mori, K.</dc:creator>
<dc:creator>Kane, L. P.</dc:creator>
<dc:date>2025-07-11</dc:date>
<dc:identifier>doi:10.1101/2025.07.08.663709</dc:identifier>
<dc:title><![CDATA[Tim-3 Promotes Early Differentiation of Tbet+ Effector T Cells During Acute Viral Infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.09.664020v1?rss=1">
<title>
<![CDATA[
ImmuneFM: Pre-training Foundation Model from Cytometry Data for Immunology Research 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.09.664020v1?rss=1</link>
<description><![CDATA[
Immunology is an essential field in the biomedicine domain, which plays an important role in oncology, vaccines, infection, etc. With the increasing amount of data available in immunology and artificial intelligence technique development, there is a need to develop data-driven AI methods in the field. However, the data from various immunology studies is very hard to integrate into an AI-ready dataset due to the lack of a standard. Moreover, independent immunology studies data lacks enough labels to train the supervised model. Motivated by these challenges, we curated a large-scale AI-ready cytometry dataset for immunology from the publicly available ImmPort portal. We design the framework to pre-train a foundation model, ImmuneFM, on the cytometry dataset. ImmuneFM can be applied to a wide range of downstream immunology diseases with fine-tuning on a limited number of labeled samples. The experiment results on eight downstream tasks demonstrate the superior performance of ImmuneFM compared to baseline deep learning and traditional methods.
]]></description>
<dc:creator>Ding, S.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Butte, A.</dc:creator>
<dc:date>2025-07-11</dc:date>
<dc:identifier>doi:10.1101/2025.07.09.664020</dc:identifier>
<dc:title><![CDATA[ImmuneFM: Pre-training Foundation Model from Cytometry Data for Immunology Research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.11.661192v1?rss=1">
<title>
<![CDATA[
A Fibroblast State Choreographs an Epithelial YAP-dependent Regenerative Program Essential to (Pre)malignancy via ECM-mediated Mechanotransduction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.11.661192v1?rss=1</link>
<description><![CDATA[
Chronic lung injury generates metaplasia which occasionally, but ominously, progresses to squamous dysplasia and squamous lung cancer. To identify mechanisms through which disrupted tissue homeostasis contributes to malignant initiation and progression, we used in vivo and in vitro heterotypic recombinant models of human bronchial epithelial cells (hBECs) and fibroblasts. We demonstrate that injury-associated TGF-{beta} signaling creates a fibroblast state dependent upon HSP47 upregulation. These fibroblasts accumulated collagen, thus elevating tissue stiffness and activating mechanosignaling that sustained YAP-dependent embryonic-like, pro-malignant activities in adjacent hBECs. This Stress/Tension-Instructive Fibroblast (STIF) state, exhibited by stressed fibroblasts in premalignant and malignant lesions across multiple cancer types, was sufficient to reprogram disease-free hBECs to metaplasia and to drive hBECs with compromised tumor suppressor function to dysplasia, yet could be inhibited and reversed. STIFs suffice to activate epithelial phenotypes reminiscent of oncogene-mediated cell transformation and induce (pre)malignancy via increased force transmission, providing novel targets for prevention.

Statement of significanceTissue injury creates a regenerative pro-tumorigenic Stress/Tension-Instructive Fibroblast (STIF) state which is sufficient to activate a YAP-dependent, pre-malignant program to induce or unmask pre-cancerous phenotypes in epithelial cells through mechanotransduction. Inhibition of STIF activity or mechanosignaling prevents metaplasia and progression to dysplasia.

HighlightsO_LITissue injury creates a pro-tumorigenic Stress/Tension-Instructive Fibroblast (STIF) state in multiple organs that precedes and persists through cancer
C_LIO_LISTIF signaling alone, working through fibroblasts and not epithelial cells, is sufficient to activate embryonic-like plasticity and induce epithelial pre-cancerous metaplastic lesions
C_LIO_LISTIFs program (pre)malignant phenotypes in adjacent epithelial cells through mechanosignaling by activating YAP prior to tumor formation
C_LIO_LIInhibiting STIFs or mechanosignaling prevents/reverts metaplasia and prevents progression to dysplasia
C_LI
]]></description>
<dc:creator>Pan, D.</dc:creator>
<dc:creator>Gascard, P.</dc:creator>
<dc:creator>Caruso, J. A.</dc:creator>
<dc:creator>Chen-Tanyolac, C.</dc:creator>
<dc:creator>Sangwan, V.</dc:creator>
<dc:creator>Bertos, N.</dc:creator>
<dc:creator>Camilleri-Broet, S.</dc:creator>
<dc:creator>Berube, J.</dc:creator>
<dc:creator>Oikonomopoulos, S.</dc:creator>
<dc:creator>Strasser, M. K.</dc:creator>
<dc:creator>Gibbs, D. L.</dc:creator>
<dc:creator>Bons, J.</dc:creator>
<dc:creator>Burton, J. B.</dc:creator>
<dc:creator>Rose, J. P.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Bai, R.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Fels-Elliott, D. R.</dc:creator>
<dc:creator>Miyara, S.</dc:creator>
<dc:creator>Alon, U.</dc:creator>
<dc:creator>Urisman, A.</dc:creator>
<dc:creator>Ragoussis, I.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:creator>Ferri, L. E.</dc:creator>
<dc:creator>Tlsty, T. D.</dc:creator>
<dc:date>2025-07-11</dc:date>
<dc:identifier>doi:10.1101/2025.07.11.661192</dc:identifier>
<dc:title><![CDATA[A Fibroblast State Choreographs an Epithelial YAP-dependent Regenerative Program Essential to (Pre)malignancy via ECM-mediated Mechanotransduction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.11.664426v1?rss=1">
<title>
<![CDATA[
KIF5A downregulation in spinal muscular atrophy links axonal regeneration defects with ALS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.11.664426v1?rss=1</link>
<description><![CDATA[
Spinal muscular atrophy (SMA) is a devastating neuromuscular disorder caused by mutations in the Survival Motor Neuron 1 (SMN1) gene, leading to decreased SMN levels and motor neuron dysfunction. SMN-restoring therapies offer clinical benefit, but the downstream molecular consequences of SMN reduction remain incompletely understood. Here, we demonstrate that SMN deficiency results in downregulation of KIF5A in human neurons and in a mouse model of SMA. We provide evidence that reduced SMN levels impair axon regeneration, which is rescued by KIF5A overexpression and that the RNA-binding protein SMN functions to stabilize KIF5A mRNA. These findings provide evidence of a molecular link between SMA and ALS pathophysiology, highlighting KIF5A as a new SMN target. Our findings suggest SMN-independent interventions targeting KIF5A could represent a complementary therapeutic approach for SMA and other motor neuron diseases.
]]></description>
<dc:creator>Akiyama, T.</dc:creator>
<dc:creator>Zeng, Y.</dc:creator>
<dc:creator>Guo, C.</dc:creator>
<dc:creator>Gautier, O.</dc:creator>
<dc:creator>Koepke, L. S.</dc:creator>
<dc:creator>Bombosch, J.</dc:creator>
<dc:creator>Sianto, O.</dc:creator>
<dc:creator>Ross, J. P.</dc:creator>
<dc:creator>Hoang, P. T.</dc:creator>
<dc:creator>Zhao, L. Y.</dc:creator>
<dc:creator>Spencer, C.</dc:creator>
<dc:creator>Monje, M.</dc:creator>
<dc:creator>Day, J. W.</dc:creator>
<dc:creator>Gitler, A. D.</dc:creator>
<dc:date>2025-07-13</dc:date>
<dc:identifier>doi:10.1101/2025.07.11.664426</dc:identifier>
<dc:title><![CDATA[KIF5A downregulation in spinal muscular atrophy links axonal regeneration defects with ALS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.13.664555v1?rss=1">
<title>
<![CDATA[
Cortical organoids reveal human-specific roles of METTL5 in neurodevelopment via regulation of CHCHD2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.13.664555v1?rss=1</link>
<description><![CDATA[
METTL5 is a conserved methyltransferase responsible for catalyzing N6-methyladenosine (m6A) modification on 18S ribosomal RNA. Human patients with mutations in METTL5 show severe microcephaly and intellectual disability, which has not been fully recapitulated in animal models. Given its emerging role in neurodevelopment, we sought to investigate METTL5 function in a human-specific context using cortical forebrain organoids derived from human induced pluripotent stem cells. We generated METTL5 knockout organoids and observed a marked delay in neural stem cell proliferation and the timing of neuronal differentiation, suggesting a critical role for METTL5 in the temporal regulation of neurogenesis. Though METTL5 methylates rRNA near the decoding center, the mechanism of this methylation remains highly contested. In our cortical organoids, broad translational changes mirror stress response rather than transcript-specific regulation of translation. Transcriptomic analysis further revealed a significant downregulation of CHCHD2, a nuclear-encoded mitochondrial gene linked to cellular energy metabolism and neurodevelopmental processes. Overexpression of CHCHD2 rescued proliferation defects of METTL5-KO neural progenitor cells, suggesting dysregulation of CHCHD2 is heavily involved in the phenotypes of METTL5 mutant patient brains. These findings highlight a previously uncharacterized link between METTL5, ribosomal RNA modification, and cellular metabolism essential for proper human brain development.
]]></description>
<dc:creator>Turkalj, E. M.</dc:creator>
<dc:creator>Kuljanishvili, M.</dc:creator>
<dc:creator>Kang, G.</dc:creator>
<dc:creator>Liu, I.</dc:creator>
<dc:creator>Ghent, C.</dc:creator>
<dc:creator>Waltier Soriano, C.</dc:creator>
<dc:creator>Keyhanvar, N.</dc:creator>
<dc:creator>Brody, D.</dc:creator>
<dc:creator>Oldham, M.</dc:creator>
<dc:creator>Vissers, C.</dc:creator>
<dc:date>2025-07-13</dc:date>
<dc:identifier>doi:10.1101/2025.07.13.664555</dc:identifier>
<dc:title><![CDATA[Cortical organoids reveal human-specific roles of METTL5 in neurodevelopment via regulation of CHCHD2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.13.663563v1?rss=1">
<title>
<![CDATA[
The Notch1 intracellular domain orchestrates mechanotransduction of fluid shear stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.13.663563v1?rss=1</link>
<description><![CDATA[
Hemodynamic shear stress regulates endothelial phenotype through activation of Notch1 signaling, yet the mechanistic basis for this activation is unclear. Here, we establish a fluid shear stress-dependent mechanism of Notch1 activation that is distinct from canonical ligand trans-endocytosis. Application of laminar flow triggers the rapid spatial polarization of full-length Notch1 heterodimers into downstream membrane microdomains. Unlike canonical transactivation, this response occurs independently of ligand redistribution, and Notch1 receptors are cis-endocytosed into the receptor-bearing cell within polarized microdomains prior to proteolytic activation. We discover that the Notch1 intracellular domain (ICD) critically orchestrates receptor polarization and proteolytic activation in response to flow but is dispensable for canonical ligand transactivation. Shear stress increases ICD interaction with annexin A2 and caveolar proteins which control Notch1 endocytosis and proteolytic compartmentalization. These findings define a flow-specific Notch1 mechanotransduction pathway linking receptor polarization and endocytosis with proteolytic activation and establish new mechanisms regulating Notch receptor activation.
]]></description>
<dc:creator>Singh, T.</dc:creator>
<dc:creator>Jacobs, K. A.</dc:creator>
<dc:creator>Polacheck, W. J.</dc:creator>
<dc:creator>Kutys, M. L.</dc:creator>
<dc:date>2025-07-14</dc:date>
<dc:identifier>doi:10.1101/2025.07.13.663563</dc:identifier>
<dc:title><![CDATA[The Notch1 intracellular domain orchestrates mechanotransduction of fluid shear stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.11.664189v1?rss=1">
<title>
<![CDATA[
Tumor-initiating genetics and therapy drive divergent molecular evolution in IDH-mutant gliomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.11.664189v1?rss=1</link>
<description><![CDATA[
Astrocytomas and oligodendrogliomas are slow-growing and treatment-sensitive IDH-mutant gliomas diagnosed at ages 30-50. Local tumor regrowth and treatment resistance is inevitable resulting in 3-10 year astrocytoma and up to >20 years oligodendroglioma survival. We sought to identify genetic changes associated with tumor evolution in response to therapy through multi-timepoint whole-genome/whole-exome sequencing of 206 IDH-mutant glioma patient samples collected through the Glioma Longitudinal Analysis (GLASS) Consortium. We validated known genomic markers of tumor progression, including hypermutation and CDKN2A homozygous deletion, and discovered novel genetic alterations that distinguish the response to treatment in astrocytomas compared to oligodendrogliomas. Point mutations in PIK3CA, PIK3R1, and NOTCH1 were newly acquired in recurrent oligodendrogliomas and associated with increased mutation rates. Focal oncogene amplifications, together with CDKN2A homozygous deletions, were associated with an increase in recurrence-specific chromosomal imbalances in astrocytomas. Mutational signature analysis revealed additional differences and detected enrichment for the SBS11, and SBS119 mutational signatures after temozolomide treatment in both IDH-glioma subtypes, whereas astrocytomas showed increased ID8 signatures after radiotherapy. These signatures suggest that the genomes of oligodendroglioma and astrocytoma adapt to the selective pressures of tumor progression and treatment in different ways. However, in both IDH-mutant glioma subtypes we observed a convergence of acquired driver gene alterations with genome-wide changes and worse patient outcomes, signaling selection of treatment-refractory clones. By identifying new prognostic markers and delineating the genomic divergence of oligodendrogliomas and astrocytomas after diagnosis, our results suggest that different DNA damage response mechanisms are engaged following chemo- and radiation therapy.
]]></description>
<dc:creator>Chowdhury, T.</dc:creator>
<dc:creator>Tesileanu, C. M. S.</dc:creator>
<dc:creator>Kocakavuk, E.</dc:creator>
<dc:creator>Johnson, K. C.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Erson-Omay, Z.</dc:creator>
<dc:creator>Heo, C.</dc:creator>
<dc:creator>Aldape, K.</dc:creator>
<dc:creator>Amin, S. B.</dc:creator>
<dc:creator>Anderson, K. J.</dc:creator>
<dc:creator>Ashley, D. M.</dc:creator>
<dc:creator>Barnholtz-Sloan, J. S.</dc:creator>
<dc:creator>Brat, D. J.</dc:creator>
<dc:creator>Brodbelt, A. R.</dc:creator>
<dc:creator>Castro, A. V.</dc:creator>
<dc:creator>Claus, E. B.</dc:creator>
<dc:creator>Connelly, J. M.</dc:creator>
<dc:creator>Costello, J.</dc:creator>
<dc:creator>Datta, I.</dc:creator>
<dc:creator>Elliott, C.</dc:creator>
<dc:creator>Finocchiaro, G.</dc:creator>
<dc:creator>French, P. J.</dc:creator>
<dc:creator>Gan, H. K.</dc:creator>
<dc:creator>Garofano, L.</dc:creator>
<dc:creator>Ghospurkar, P. L.</dc:creator>
<dc:creator>Golebiewska, A.</dc:creator>
<dc:creator>Gundla, P. S.</dc:creator>
<dc:creator>Hermes, B.</dc:creator>
<dc:creator>Hong, C.</dc:creator>
<dc:creator>Hoogstrate, Y.</dc:creator>
<dc:creator>Horbinski, C.</dc:creator>
<dc:creator>Huse, J. T.</dc:creator>
<dc:creator>Iavarone, A.</dc:creator>
<dc:creator>Karadag, C.</dc:creator>
<dc:creator>Khasraw, M.</dc:creator>
<dc:creator>Kouwenhoven, M. C.</dc:creator>
<dc:creator>LaViolette, P. S.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Lowman, A.</dc:creator>
<dc:creator>McCortney, K.</dc:creator>
<dc:creator>Moon, H.-E.</dc:creator>
<dc:creator>Na</dc:creator>
<dc:date>2025-07-15</dc:date>
<dc:identifier>doi:10.1101/2025.07.11.664189</dc:identifier>
<dc:title><![CDATA[Tumor-initiating genetics and therapy drive divergent molecular evolution in IDH-mutant gliomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.10.664265v1?rss=1">
<title>
<![CDATA[
Auto-MedCalc: Automated Biomarkers Discovery and Risk Score Generation with AI Agents 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.10.664265v1?rss=1</link>
<description><![CDATA[
Identifying biomarkers and generating risk scores are usually essential tasks in many biomedicine and clinical scenarios. However, this is a highly hypothesis-driven and experience-dependent process requiring extensive experiments as well. How to discover biomarkers from multi-modality, multi-source data and generate risk scores with higher precision motivates us to design the framework Auto-MedCalc, a data-driven pipeline to automatically identify biomarkers and generate risk scores for diagnosis. Auto-MedCalc is a multi-agent AI system with large language models and computational tools. Three representative studies of transplantation from the public ImmPort data portal are used for the experiment. Auto-MedCalc achieved 0.93, 0.88, and 0.88 ROC-AUC on the rejection prediction after kidney, liver, and heart transplants, surpassing the best expert-designed medical calculators by 50%, 4.7%, and 39%, respectively. Auto-MedCalc also validated the previously found human biomarkers and discovered new biomarkers, {beta}-Glucuronidase and Alpha-1-microglobulin, in heart transplants, which hadnt attracted much attention before. The superior performance from experiments shows that Auto-MedCalc can be used for both clinical applications and scientific discovery.
]]></description>
<dc:creator>Ding, S.</dc:creator>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Butte, A.</dc:creator>
<dc:date>2025-07-16</dc:date>
<dc:identifier>doi:10.1101/2025.07.10.664265</dc:identifier>
<dc:title><![CDATA[Auto-MedCalc: Automated Biomarkers Discovery and Risk Score Generation with AI Agents]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.15.664547v1?rss=1">
<title>
<![CDATA[
Programmable translational inhibition by a molecular glue-oligonucleotide conjugate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.15.664547v1?rss=1</link>
<description><![CDATA[
Selective inhibition of mRNA translation is a promising strategy for modulating the activity of disease-associated genes, yet achieving both high potency and specificity remains challenging. Rocaglamide A (RocA), a molecular glue, inhibits translation by clamping eIF4A onto polypurine motifs found in many transcripts, thereby limiting RocAs specificity. Here, we developed RocASO, a chemical conjugate that links RocA to an antisense oligonucleotide (ASO) capable of base-pairing with defined mRNA sequences, thus directing RocAs clamping mechanism to chosen targets and enhancing overall potency and specificity. We show that RocASOs are compatible with various types of ASO modalities, including gapmers that induce the degradation of target RNAs. RocASOs were designed to effectively knock down endogenous genes (PTGES3, HSPA1B) and SARS-CoV-2 viral RNA, the latter conferring potent antiviral activity in cells. These findings establish RocASO as a versatile platform for programmable translational inhibition with therapeutic potential.
]]></description>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Kovalski, J. R.</dc:creator>
<dc:creator>Zapatero-Belinchon, F. J.</dc:creator>
<dc:creator>Bennett, M.</dc:creator>
<dc:creator>Kuzuoglu-Öztürk, D.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:creator>Ruggero, D.</dc:creator>
<dc:creator>Lou, K.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:date>2025-07-16</dc:date>
<dc:identifier>doi:10.1101/2025.07.15.664547</dc:identifier>
<dc:title><![CDATA[Programmable translational inhibition by a molecular glue-oligonucleotide conjugate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.15.664994v1?rss=1">
<title>
<![CDATA[
Primary cilia and BBS4 are required for postnatal pituitary development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.15.664994v1?rss=1</link>
<description><![CDATA[
Primary cilia orchestrate several signaling pathways, and their disruption results in pleiotropic disorders called ciliopathies. Bardet Beidl syndrome (BBS), one such ciliopathy, provides insights into cilia function in many tissues. Using a mouse model of BBS, Bbs4 knockout (Bbs4-/-), and conditional deletion of pituitary cilia, we found that adult Bbs4-/- and cilia-less pituitaries are hypoplastic and have increased gonadotroph populations. The developing Bbs4-/- pituitary experiences mildly reduced Hedgehog signaling. Isolated Bbs4-/- pituitary stem cells exhibited reduced Hedgehog signal responsiveness and expression of stem cell markers. These data demonstrate that cilia and BBS function are necessary for pituitary growth. We propose that altered pituitary cilia-mediated patterning contribute to physiological features of ciliopathies such as BBS.

Summary statementA mouse model of BBS reveals that ciliary Hedgehog signaling is necessary for postnatal pituitary development.
]]></description>
<dc:creator>Brewer, K. M.</dc:creator>
<dc:creator>Brewer, K. K.</dc:creator>
<dc:creator>Richardson, N. C.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:creator>Berbari, N. F.</dc:creator>
<dc:creator>Konjikusic, M. J.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.15.664994</dc:identifier>
<dc:title><![CDATA[Primary cilia and BBS4 are required for postnatal pituitary development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.17.664991v1?rss=1">
<title>
<![CDATA[
Tracking tau and cellular responses in human iPSC-microglia from uptake to seedable secretion in extracellular vesicles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.17.664991v1?rss=1</link>
<description><![CDATA[
The templated spread of tau aggregates in tauopathies has been attributed to neuron-to- neuron spread, but microglia have also been implicated through mouse studies. Here we examine in detail the uptake, processing, release and seeding of tau using human iPS- derived microglia (iMGL). We show that tau is taken up by iMGL via LRP1 and heparan sulfate proteoglycans, with a role for LRRK2 in LRP1 trafficking, and that phagocytosed fibrils can escape into the cytoplasm. Monomeric tau has minimal effects on iMGL, but recombinant or brain-derived tau fibrils induce a shift towards chemokine and interferon response subtypes, alongside downregulation of homeostatic and MHC genes. Endogenous tau protein is undetectable in iMGL, and monomeric internalised tau is digested to completion, but fibrillar tau is more resistant to degradation and becomes phosphorylated on two specific residues. Finally, fibrillar tau is released by iMGL, visualized within extracellular vesicles by cryo-EM, and can seed tau aggregation in downstream neurons.
]]></description>
<dc:creator>Karabova, M. K.</dc:creator>
<dc:creator>del Ser Badia, A.</dc:creator>
<dc:creator>Hedegaard, A.</dc:creator>
<dc:creator>Washer, S. J.</dc:creator>
<dc:creator>Baykam, Z.</dc:creator>
<dc:creator>O'Brien, D. P.</dc:creator>
<dc:creator>Vendrell, I.</dc:creator>
<dc:creator>Hester, S. S.</dc:creator>
<dc:creator>Fischer, R.</dc:creator>
<dc:creator>Johnson, E.</dc:creator>
<dc:creator>Melia, C. E.</dc:creator>
<dc:creator>Matthews-Palmer, T. R. S.</dc:creator>
<dc:creator>Matadeen, R.</dc:creator>
<dc:creator>Santambrogio, A.</dc:creator>
<dc:creator>Metrick, M.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Keeling, S.</dc:creator>
<dc:creator>Cheam, K.</dc:creator>
<dc:creator>McEwan, W. A.</dc:creator>
<dc:creator>Kosik, K. S.</dc:creator>
<dc:creator>Day, T. A.</dc:creator>
<dc:creator>James, W. S.</dc:creator>
<dc:creator>Cowley, S. A.</dc:creator>
<dc:date>2025-07-21</dc:date>
<dc:identifier>doi:10.1101/2025.07.17.664991</dc:identifier>
<dc:title><![CDATA[Tracking tau and cellular responses in human iPSC-microglia from uptake to seedable secretion in extracellular vesicles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.17.665438v1?rss=1">
<title>
<![CDATA[
Programmable microparticles rewire CAR signaling to enable super-physiological expansion of human T cells in vitro 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.17.665438v1?rss=1</link>
<description><![CDATA[
T cell proliferative capacity and persistence critically determine the therapeutic success of chimeric antigen receptor (CAR) T cells. However, it remains unknown if and how human CAR-T cells can be externally programmed to reach maximal proliferative capacity. Here, we use programmable PLGA microparticles functionalized with CAR-antigens and CD28-costimulatory antibodies (CAREp) to repeatedly stimulate human CD8+ CAR-T cells in vitro. CAREp-stimulated CAR-T cells expanded continuously for over 100 days--versus [~]30 days with tumor cell stimulation--and achieved up to 1018-fold cumulative expansion, greatly surpassing CD3/28-Dynabeads. Early-phase transcriptomic responses-- upregulation of DNA repair, cell cycle, telomere maintenance, and mitochondrial pathways--aligned with long-term outcomes: massive proliferation, telomere stability, robust respiration, and preserved progenitor phenotype by single-cell sequencing. Differentiation and exhaustion signals were broadly suppressed. Transient telomerase activity further supported physiologic expansion. These findings demonstrate that nanoscale-controlled extracellular cues can rewire intracellular signaling to drive durable, super-physiological expansion of functional CAR-T cells.
]]></description>
<dc:creator>Zeng, Q.</dc:creator>
<dc:creator>Flemming, L.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Mazumder, T.</dc:creator>
<dc:creator>Hammerlindl, H.</dc:creator>
<dc:creator>Allen, G. M.</dc:creator>
<dc:creator>Almeida, R.</dc:creator>
<dc:creator>Williams, J. Z.</dc:creator>
<dc:creator>Hernandez-Lopez, R. A.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Lim, W. A.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:creator>Desai, T.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:date>2025-07-22</dc:date>
<dc:identifier>doi:10.1101/2025.07.17.665438</dc:identifier>
<dc:title><![CDATA[Programmable microparticles rewire CAR signaling to enable super-physiological expansion of human T cells in vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.22.665939v1?rss=1">
<title>
<![CDATA[
Mapping mesenchymal diversity in the developing human intestine and organoids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.22.665939v1?rss=1</link>
<description><![CDATA[
The organization of diverse mesenchymal populations during human intestinal development is critical for tissue architecture and function yet remains poorly defined. To construct a comprehensive, tissue-scale map of the developing human small intestine, we leveraged single-cell RNA-sequencing data to build a custom Xenium spatial transcriptomics gene panel covering the diversity of cell types in the human intestine. Analysis was focused on the developing mesenchyme populations (also referred to as fibroblasts or stroma) given the lack of spatiotemporal information about these cell populations. We defined 5 broad mesenchymal populations occupying discrete anatomical locations within the lamina propria and submucosa - the subepithelial cells (SEC), lamina propria fibroblasts (LPF), submucosal fibroblasts (SMF), smooth muscle cells (SMC), and CXCL13+ fibroblasts. Our data reveal dynamic spatial remodeling of fibroblast communities during development and establish molecular markers to distinguish these populations. We leverage this high-resolution atlas to benchmark pluripotent stem cell-derived human intestinal organoids and to demonstrate how this foundational resource can be used to dissect intestinal stromal signaling in a spatial manner, with broad implications for modeling development, regeneration, and disease.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=177 SRC="FIGDIR/small/665939v1_ufig1.gif" ALT="Figure 1">
View larger version (69K):
org.highwire.dtl.DTLVardef@46299borg.highwire.dtl.DTLVardef@55a014org.highwire.dtl.DTLVardef@180a104org.highwire.dtl.DTLVardef@1cb42b2_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LIA spatial atlas of fibroblast heterogeneity in the human intestine
C_LIO_LIPopulations include subepithelial, submucosal, lamina propria, and CXCL13+ fibroblasts
C_LIO_LIFibroblast position in the developing intestine is maintained into adulthood
C_LIO_LIOrganoids largely recreate the spatial organization of the human intestine
C_LI
]]></description>
<dc:creator>Johnson, K. F.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Tsai, Y.-H.</dc:creator>
<dc:creator>Wu, A.</dc:creator>
<dc:creator>Clark, S. G.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Zwick, R.</dc:creator>
<dc:creator>Glass, I.</dc:creator>
<dc:creator>Walton, K. D.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Spence, J.</dc:creator>
<dc:date>2025-07-22</dc:date>
<dc:identifier>doi:10.1101/2025.07.22.665939</dc:identifier>
<dc:title><![CDATA[Mapping mesenchymal diversity in the developing human intestine and organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.19.665688v1?rss=1">
<title>
<![CDATA[
Receptor-tethered orthogonal IL-2 enhances regulatory T cell therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.19.665688v1?rss=1</link>
<description><![CDATA[
Regulatory T cell (Treg) therapy is an emerging platform for controlling immune overactivation. Efficacy of Treg therapy is limited by the poor persistence of infused Tregs due to insufficient IL-2, which is essential for Treg survival and function. IL-2 activates many immune cells besides Tregs, imposing a challenge for the selective provision of IL-2 to infused Tregs. In this study, we found infusions of orthogonal (ortho) IL-2 failed to enhance Tregs engineered with a corresponding orthoIL-2 receptor (IL-2R) in a mouse model of autoimmune diabetes. We then developed a receptor-tethered orthoIL-2 by optimizing the combination of IL-2, IL-2R, and the linker connecting them to achieve autocrine signaling selectively in engineered Tregs. Tregs expressing the tethered orthoIL-2 showed autocrine IL-2 signaling in vitro, enhanced CD25, CTLA-4, and Foxp3 expression, persisted without exogenous IL-2 in vivo, and prevented autoimmune diabetes using as few as 2,000 Tregs. Knocking the tethered orthoIL-2 construct into the Foxp3 locus enabled Treg-specific expression, with the additional benefit of positively reinforcing tethered orthoIL-2 expression through the activation of the Foxp3 gene by enhanced IL-2 signaling. Together, these results illustrate a safe and effective cell-engineering solution for overcoming Tregs dependency on exogenous IL-2, thereby achieving superior therapeutic efficacy.
]]></description>
<dc:creator>Indart, A.</dc:creator>
<dc:creator>Lyu, H.</dc:creator>
<dc:creator>Nguyen, V.</dc:creator>
<dc:creator>Rosenthal, W.</dc:creator>
<dc:creator>Jang, S.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Jude, K.</dc:creator>
<dc:creator>Su, L.</dc:creator>
<dc:creator>Garcia, K. C.</dc:creator>
<dc:creator>Bluestone, J. A.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:date>2025-07-23</dc:date>
<dc:identifier>doi:10.1101/2025.07.19.665688</dc:identifier>
<dc:title><![CDATA[Receptor-tethered orthogonal IL-2 enhances regulatory T cell therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.22.666189v1?rss=1">
<title>
<![CDATA[
Canonical and phosphoribosyl ubiquitination coordinate to stabilize a proteinaceous structure surrounding the Legionella-containing vacuole 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.22.666189v1?rss=1</link>
<description><![CDATA[
Legionella pneumophila (L.p.), an intracellular bacterial pathogen, hijacks the ubiquitin signaling network of its eukaryotic host cells to establish infection. Two families of L.p. secreted ubiquitin ligases are instrumental in the maturation of the Legionella-containing vacuole (LCV): the SidC/SdcA family, which catalyzes canonical ubiquitination, and the SidE family, which bypasses the E1-E2-E3 enzymatic cascade and directly conjugates ubiquitin to a target through a phosphoribosyl (PR) linkage. Here, we demonstrate that the coordinated activities of these two effector families generate a hyperstable, ubiquitin-rich structure surrounding the LCV. We propose a model in which an initial wave of SidC/SdcA-mediated canonical ubiquitination around the LCV is further modified by SidE family-driven PR-ubiquitination, resulting in a detergent-resistant "cloud". The "cloud" is transient, breaking down as infection progresses, suggesting that L.p. reshapes the properties of the proteinaceous shell surrounding the vacuole to meet changing needs throughout its intracellular lifecycle. This unusual structure likely stabilizes and protects the LCV, shielding it from host defense mechanisms during early infection. Our findings reveal cellular consequences of effector interplay during infection and provide a foundation for future studies into the structure and function of the proteinaceous "cloud" surrounding the LCV.
]]></description>
<dc:creator>Steinbach, A.</dc:creator>
<dc:creator>Mokkapati, C.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:date>2025-07-23</dc:date>
<dc:identifier>doi:10.1101/2025.07.22.666189</dc:identifier>
<dc:title><![CDATA[Canonical and phosphoribosyl ubiquitination coordinate to stabilize a proteinaceous structure surrounding the Legionella-containing vacuole]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.21.665156v1?rss=1">
<title>
<![CDATA[
Integrated single-nuclei and spatial transcriptomic profiling of human sacrococcygeal teratomas reveals heterogeneity in cellular composition and X-chromosome inactivation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.21.665156v1?rss=1</link>
<description><![CDATA[
Sacrococcygeal teratomas (SCTs) are the most common neonatal tumors, yet their cellular origins, clinical stratification, and sex bias-occurring three times more in XX than XY individuals--remain poorly understood. To address these gaps, we examined six postnatal (one male and five female) and two prenatal (both female) SCTs by single nuclei RNA-seq and spatial transcriptomics. We identified five broad cellular lineages in SCTs: stroma, epithelia, endothelia, neuroectoderm, and immune. The transcriptomes and lineage compositions showed significant heterogeneity, which offer a framework for future molecular stratification. SCTs are thought to originate from and be propagated by pluripotent cells, notably however, we did not detect these populations. Among female tumors, a subset of cells exhibited biallelic expression of X-linked genes, consistent with X-inactivation failure or reactivation of the once inactivated X-chromosome. These biallelic cells were enriched for developmental and neuronal programs, whereas cells with single-allelic X-chromosome preferentially expressed immune-related genes. Biallelic X-chromosome activation, which can occur only in female cells, may result in transcriptomic features that favor survival of tumor cells, contributing to the sex bias of SCTs. Our findings reveal a link between X-chromosome inactivation and SCT cell identity, suggesting that X-dosage dysregulation may influence SCT heterogeneity and immune landscape.
]]></description>
<dc:creator>Rojas, E. J.</dc:creator>
<dc:creator>Giannikou, K.</dc:creator>
<dc:creator>Huang, B. J.</dc:creator>
<dc:creator>Cho, S.-J.</dc:creator>
<dc:creator>Cordero, M.</dc:creator>
<dc:creator>Pena, D.</dc:creator>
<dc:creator>Vu, L.</dc:creator>
<dc:creator>Bagrodia, A.</dc:creator>
<dc:creator>Derderian, S. C.</dc:creator>
<dc:creator>MacKenzie, T. C.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:date>2025-07-24</dc:date>
<dc:identifier>doi:10.1101/2025.07.21.665156</dc:identifier>
<dc:title><![CDATA[Integrated single-nuclei and spatial transcriptomic profiling of human sacrococcygeal teratomas reveals heterogeneity in cellular composition and X-chromosome inactivation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.21.665987v1?rss=1">
<title>
<![CDATA[
Hippocampal input-driven plasticity of prefrontal interneurons reveals a circuit basis for impaired spatial working memory. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.21.665987v1?rss=1</link>
<description><![CDATA[
Dynamic functional connectivity between the ventral hippocampus (vHPC) and medial prefrontal cortex (mPFC) is essential for spatial working memory (SWM). Interactions between vHPC projections and mPFC interneurons, and their plasticity, are uniquely positioned to influence SWM, yet the nature of these interactions remains unclear. Here, we combined in vivo optical stimulation of vHPC inputs to mPFC with calcium recordings of discrete mPFC interneuron populations in mice, revealing class-specific response profiles and plasticity. Repeated vHPC input stimulation persistently depressed activity in vasoactive intestinal peptide (VIP)-expressing interneurons and potentiated activity in somatostatin-expressing interneurons. Ex vivo whole-cell electrophysiology and computational modeling revealed that these divergent effects likely arise from a primary weakening of monosynaptic vHPC input onto VIP interneurons. Leveraging this plasticity to inform the circuit interactions that support SWM, we found that mice with prior vHPC input stimulation displayed elevated VIP interneuron activity during the delay epoch in early SWM task training, and this enhanced activity correlated with poorer training performance. Accordingly, mice modeling the schizophrenia-predisposing 22q11.2 deletion syndrome with known SWM learning deficits recapitulated this aberrant VIP interneuron activity profile and showed reduced vHPC targeting of mPFC VIP interneurons. Together, these findings reveal novel cell-type-specific plasticity in cognition-supporting circuits and illustrate how reweighting of inputs to VIP interneurons may contribute to working memory dysfunction.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=190 SRC="FIGDIR/small/665987v1_ufig1.gif" ALT="Figure 1">
View larger version (61K):
org.highwire.dtl.DTLVardef@29a326org.highwire.dtl.DTLVardef@1964208org.highwire.dtl.DTLVardef@1c94e4corg.highwire.dtl.DTLVardef@1336edb_HPS_FORMAT_FIGEXP  M_FIG C_FIG IN BRIEFSilverstein et al. investigated how ventral hippocampus (vHPC) inputs to medial prefrontal cortex (mPFC) interact with mPFC interneuron populations to support spatial working memory (SWM) in mice. They show that repeated vHPC input stimulation that reduces vHPC drive onto vasoactive intestinal peptide (VIP) interneurons also elevates delay-related VIP interneuron activity in early SWM task training, and this enhanced activity correlates with poorer training performance. Mice modelling the 22q11.2 deletion syndrome--known to have SWM learning deficits--recapitulate this altered VIP interneuron activity and show reduced vHPC input targeting of VIP interneurons.

HIGHLIGHTSO_LIRepeated stimulation of vHPC inputs to mPFC persistently depresses VIP interneuron activity and enhances SST interneuron activity in mice
C_LIO_LIBlunted monosynaptic drive onto mPFC VIP interneurons by repeated vHPC input stimulation plausibly reproduces in vivo activity changes
C_LIO_LIStimulated mice show heightened VIP interneuron activity during the delay epoch in early SWM task training that correlates with poorer training performance
C_LIO_LIMice modeling the 22q11.2 deletion syndrome with known SWM task learning deficits mirror this aberrant VIP interneuron activity profile and show reduced vHPC input targeting of VIP interneurons
C_LI
]]></description>
<dc:creator>Silverstein, S. E.</dc:creator>
<dc:creator>Clarity, T. T.</dc:creator>
<dc:creator>Deshpande, M. S.</dc:creator>
<dc:creator>Vaughan, E.</dc:creator>
<dc:creator>Novik, S.</dc:creator>
<dc:creator>Yarur, H. E.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Tsai, V. S.</dc:creator>
<dc:creator>Ye, R.</dc:creator>
<dc:creator>Mikofsky, R. M.</dc:creator>
<dc:creator>Hsiang, M.</dc:creator>
<dc:creator>Bauman, A.</dc:creator>
<dc:creator>Loewinger, G.</dc:creator>
<dc:creator>Pereira, F.</dc:creator>
<dc:creator>Morales, M.</dc:creator>
<dc:creator>Sohal, V. S.</dc:creator>
<dc:creator>Tejeda, H.</dc:creator>
<dc:creator>Gordon, J. A.</dc:creator>
<dc:creator>Kupferschmidt, D. A.</dc:creator>
<dc:date>2025-07-24</dc:date>
<dc:identifier>doi:10.1101/2025.07.21.665987</dc:identifier>
<dc:title><![CDATA[Hippocampal input-driven plasticity of prefrontal interneurons reveals a circuit basis for impaired spatial working memory.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.21.664988v1?rss=1">
<title>
<![CDATA[
An open-source pipeline for calcium imaging and all-optical physiology in human stem cell-derived neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.21.664988v1?rss=1</link>
<description><![CDATA[
High-throughput profiling of neuronal activity at single-cell resolution is essential for advancing our understanding of brain function, enabling large-scale functional screens, and modeling neurological disorders. However, existing approaches are limited by scalability, manual data processing, and variability, thus restricting their ability to detect disease-associated phenotypes. Here, we present a scalable, open-source platform that integrates optogenetic stimulation, calcium imaging, automated data acquisition, and a fully integrated analysis pipeline. By combining spontaneous and evoked activity profiling, the system enables robust quantification of dynamic neuronal responses across hundreds of stem cell-derived human neurons and multiple timepoints, facilitating phenotyping at both cellular and network levels. We validated the platform by recapitulating established activity phenotypes in neurodevelopmental disorders including CDKL5 Deficiency Disorder and SSADH deficiency. In addition, we generated CRISPR-Cas9 knock-in human induced pluripotent stem cell (hiPSC) lines stably expressing the genetically encoded calcium indicator GCaMP6s to model network dysfunction in Tuberous Sclerosis Complex (TSC). Using this system, we further demonstrated functional rescue of the altered neuronal activity observed in the TSC following pharmacological intervention. By linking single-cell dynamics to population-level phenotypes, this framework provides a powerful and broadly applicable tool for disease modeling, mechanistic studies, and therapeutic screening across a range of neurological disorders.
]]></description>
<dc:creator>Afshar-Saber, W.</dc:creator>
<dc:creator>Gasparoli, F. M.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Teaney, N. A.</dc:creator>
<dc:creator>Lalani, L.</dc:creator>
<dc:creator>Srinivasan, G.</dc:creator>
<dc:creator>Whye, D.</dc:creator>
<dc:creator>Buttermore, E.</dc:creator>
<dc:creator>Winden, K. D.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Sahin, M.</dc:creator>
<dc:date>2025-07-25</dc:date>
<dc:identifier>doi:10.1101/2025.07.21.664988</dc:identifier>
<dc:title><![CDATA[An open-source pipeline for calcium imaging and all-optical physiology in human stem cell-derived neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.21.666016v1?rss=1">
<title>
<![CDATA[
Single-Cell Profiling Reveals Altered Endometrial Cellular Features Across the Menstrual Cycle in Endometriosis Patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.21.666016v1?rss=1</link>
<description><![CDATA[
Endometriosis is a chronic, estrogen-dependent condition affecting over 190 million women globally, characterized by the ectopic presence of endometrial-like tissue that leads to inflammation, pain, and infertility. Despite its prevalence, the pathogenesis of endometriosis remains poorly understood. Here, we present a comprehensive single-cell transcriptomic atlas comprising 228,000 cells derived from 43 eutopic endometrial biopsies from patients with endometriosis, fibroid controls and healthy controls, sampled across the menstrual cycle. This analysis reveals previously uncharacterized subpopulations of endometrial fibroblasts and epithelial cells undergoing epithelial-mesenchymal transition, alongside disrupted immune cell communication networks. Comparative gene expression profiling implicates oxidative stress, aberrant cell migration, and dysregulated apoptosis as central features of the disease state. These findings suggest that endometriosis alters eutopic endometrial homeostasis, with potential consequences for fertility, regeneration, and disease progression. Our dataset provides a valuable resource for biomarker discovery and identifies candidate therapeutic targets aimed at restoring endometrial function and alleviating symptoms in affected individuals.
]]></description>
<dc:creator>Almonte-Loya, A.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Houshdaran, S.</dc:creator>
<dc:creator>Tang, X.</dc:creator>
<dc:creator>Flynn, E.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:creator>Kushnoor, D.</dc:creator>
<dc:creator>Nezhat, C.</dc:creator>
<dc:creator>Vilella, F.</dc:creator>
<dc:creator>Simon, C.</dc:creator>
<dc:creator>Vo, K. C.</dc:creator>
<dc:creator>Casillas, A.</dc:creator>
<dc:creator>Khan, U.</dc:creator>
<dc:creator>Serras, C. P.</dc:creator>
<dc:creator>Oskotsky, T. T.</dc:creator>
<dc:creator>Huang, D.</dc:creator>
<dc:creator>Vallve-Juanico, J.</dc:creator>
<dc:creator>Irwin, J.</dc:creator>
<dc:creator>Combes, A.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Fragiadakis, G.</dc:creator>
<dc:creator>Giudice, L.</dc:creator>
<dc:date>2025-07-25</dc:date>
<dc:identifier>doi:10.1101/2025.07.21.666016</dc:identifier>
<dc:title><![CDATA[Single-Cell Profiling Reveals Altered Endometrial Cellular Features Across the Menstrual Cycle in Endometriosis Patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.22.666173v1?rss=1">
<title>
<![CDATA[
Methylation variability and LINE-1 activation in multiple myeloma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.22.666173v1?rss=1</link>
<description><![CDATA[
Multiple myeloma (MM) is a type of hematological cancer that arises from uncontrolled proliferation of plasma cells. In addition to frequent genetic mutations, malignant plasma cells are characterized by alterations to the epigenome. Myeloma cells display a genome-wide loss of DNA methylation and a corresponding increase in  active chromatin modifications. The dramatic epigenetic remodeling that occurs in cancer genomes is associated with loss of silencing at transposable elements, which can impact genome regulation. Through paired epigenome and transcriptome profiling of patient derived MM samples, we have found that loss of DNA methylation in MM genomes results in the formation of partially methylated domains that are variable across patients. This loss of DNA methylation coincides with the expression of hundreds of transcripts driven by LINE-1 (L1) retrotransposons that are epigenetically silenced in normal cells. MM samples can be stratified based on L1 activity with distinct gene expression signatures. The high L1 samples are characterized by a more proliferative, less differentiated state as well as inhibition of interferon and genome defense pathways. Several L1 promoters generate chimeric transcripts with adjacent oncogenes. We further find that KRAB-zinc finger proteins (KZFPs) that are responsible for the epigenetic silencing of L1s have abnormally low abundance in MM samples with high L1 activity. These results indicate that cell proliferation in MM is associated with a loss of KZFP expression and activation of L1 elements.

SignificanceEpigenome and transcriptional profiling of patient-derived multiple myeloma samples reveals heterogeneous loss of DNA methylation, reduction of KZFP expression and LINE-1 activation as a clinical indicator of progression.
]]></description>
<dc:creator>Wan, Q.</dc:creator>
<dc:creator>Leung, A. Y.</dc:creator>
<dc:creator>Bhandari, M. V.</dc:creator>
<dc:creator>Kato, H.</dc:creator>
<dc:creator>Song, J. Y.</dc:creator>
<dc:creator>Schones, D. E.</dc:creator>
<dc:date>2025-07-26</dc:date>
<dc:identifier>doi:10.1101/2025.07.22.666173</dc:identifier>
<dc:title><![CDATA[Methylation variability and LINE-1 activation in multiple myeloma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.25.665779v1?rss=1">
<title>
<![CDATA[
Targeting RUNX1 in Macrophages Facilitates Cardiac Recovery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.25.665779v1?rss=1</link>
<description><![CDATA[
Despite advances in disease treatment, our understanding of how damaged organs recover and the mechanisms governing this process remain poorly defined. Here, we mapped the transcriptional and regulatory landscape of human cardiac recovery using single cell multiomics. Macrophages emerged as the most reprogrammed cell type. Deep learning identified the transcription factor RUNX1 as a key regulator of this process. Macrophage-specific Runx1 deletion recapitulated the human cardiac recovery phenotype in a chronic heart failure model. Runx1 deletion reprogrammed macrophages to a reparative phenotype, reduced fibrosis, and promoted cardiomyocyte adaptation. RUNX1 chromatin profiling revealed a conserved regulon that diminished during recovery. Mechanistically, the epigenetic reader BRD4 controlled Runx1 expression in macrophages. Chromatin activity mapping, combined with CRISPR perturbations, identified the precise regulatory element governing Runx1 expression. Therapeutically, small molecule Runx1 inhibition was sufficient to promote cardiac recovery. Our findings uncover a druggable RUNX1 epigenetic mechanism that orchestrates recovery of heart function.
]]></description>
<dc:creator>Amrute, J. M.</dc:creator>
<dc:creator>Zhu, A.</dc:creator>
<dc:creator>Pai, Y.-L.</dc:creator>
<dc:creator>Hector-Greene, M.</dc:creator>
<dc:creator>An, Y.</dc:creator>
<dc:creator>Lim, K. R.</dc:creator>
<dc:creator>Sheth, M.</dc:creator>
<dc:creator>Padmanabhan, A.</dc:creator>
<dc:creator>Lee, C. Y.</dc:creator>
<dc:creator>Yamawaki, T.</dc:creator>
<dc:creator>Sicklinger, F.</dc:creator>
<dc:creator>Hartmann, N.</dc:creator>
<dc:creator>Bredemeyer, A.</dc:creator>
<dc:creator>Lee, C. J. M.</dc:creator>
<dc:creator>Xu, V.</dc:creator>
<dc:creator>Bell, L.</dc:creator>
<dc:creator>Harmon, T.</dc:creator>
<dc:creator>Shin, H.</dc:creator>
<dc:creator>Parvathaneni, A.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Dutta, A. K.</dc:creator>
<dc:creator>Pruitt, D.</dc:creator>
<dc:creator>Barreda, J.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Shroff, U. N.</dc:creator>
<dc:creator>Nadadur, R.</dc:creator>
<dc:creator>Nigro, J.</dc:creator>
<dc:creator>Weinheimer, C. J.</dc:creator>
<dc:creator>Kovacs, A.</dc:creator>
<dc:creator>Cui, J.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Li, C.-M. K.</dc:creator>
<dc:creator>Kreisel, D.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Foo, R. S.-Y.</dc:creator>
<dc:creator>Schneider, R. K.</dc:creator>
<dc:creator>Engreitz, J.</dc:creator>
<dc:creator>Mann, D.</dc:creator>
<dc:creator>Rulifson, I.</dc:creator>
<dc:creator>Jackson, S.</dc:creator>
<dc:creator>Ason, B.</dc:creator>
<dc:creator>Kramann, R.</dc:creator>
<dc:creator>Drakos, S. G.</dc:creator>
<dc:creator>Leuschner, F.</dc:creator>
<dc:creator>Alexanian, M.</dc:creator>
<dc:creator>Lavine, K. J.</dc:creator>
<dc:date>2025-07-26</dc:date>
<dc:identifier>doi:10.1101/2025.07.25.665779</dc:identifier>
<dc:title><![CDATA[Targeting RUNX1 in Macrophages Facilitates Cardiac Recovery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.23.666334v1?rss=1">
<title>
<![CDATA[
Aggressive Cholesterol Lowering Normalizes Atherosclerosis Regression in Jak2V617F Clonal Hematopoiesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.23.666334v1?rss=1</link>
<description><![CDATA[
BackgroundThe Jak2V617F (Jak2VF) mutation is an important cause of both clonal hematopoiesis of indeterminate potential (CHIP) and myeloproliferative neoplasms (MPN). Mouse models of Jak2VF CHIP and MPN show accelerated atherosclerosis progression, driven by macrophage inflammasome activation. We undertook the present study to assess the hypothesis that ongoing inflammation would impede atherosclerosis resolution in Jak2VF mice.

Methods and ResultsChimeric Jak2VF/WT or control WT/WT bone marrow was transplanted into Ldlr-/-mice and, following 13-16 weeks of Western diet-induced atherosclerosis progression, cholesterol was lowered either moderately (to 200-300 mg/dl) or markedly (to 100 mg/dl). With moderate cholesterol lowering, there was impaired resolution of lesions in Jak2VFMPN mice compared to controls. However, with marked cholesterol lowering, progression of lesions was halted in both Jak2VF MPN and control mice while macrophage burden was decreased and lesional collagen was increased similarly in Jak2VFMPN and control mice.

Two mechanisms of low-density lipoprotein (LDL) lowering-induced suppression of inflammation in plaques were implicated: 1) reversal of increased proliferation, DNA damage and Absent in Melanoma 2 (AIM2) inflammasome activation specifically in Jak2VF macrophages and 2) markedly increased macrophage triggering receptor expressed on myeloid cells 2 (TREM2), c-myc expressing macrophages in both Jak2VF and control mice.

ConclusionsAggressive LDL lowering reverses inflammasome activation and induces pro-resolving changes in macrophages in Jak2VF MPN, halting atherosclerosis progression and promoting features of plaque stabilization. These findings suggest that aggressive LDL cholesterol lowering could reverse atherosclerotic cardiovascular disease (ACVD) risk in individuals with JAK2VFCHIP or MPN.
]]></description>
<dc:creator>Hardaway, B. D.</dc:creator>
<dc:creator>Fidler, T. P.</dc:creator>
<dc:creator>Tavallaie, M.</dc:creator>
<dc:creator>Avrampou, K.</dc:creator>
<dc:creator>Hsu, C.-C.</dc:creator>
<dc:creator>Schiavone, S.</dc:creator>
<dc:creator>Xiao, T.</dc:creator>
<dc:creator>Wang, N.</dc:creator>
<dc:creator>Tall, A. R.</dc:creator>
<dc:date>2025-07-27</dc:date>
<dc:identifier>doi:10.1101/2025.07.23.666334</dc:identifier>
<dc:title><![CDATA[Aggressive Cholesterol Lowering Normalizes Atherosclerosis Regression in Jak2V617F Clonal Hematopoiesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.23.666437v1?rss=1">
<title>
<![CDATA[
Metastasis-associated wound repair promotes reciprocal activation of the lung epithelium and breast cancer metastases during outgrowth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.23.666437v1?rss=1</link>
<description><![CDATA[
When tumor cells colonize distant organs during metastasis, they interact extensively with surrounding cells. These interactions often change the behavior of surrounding cell populations which collectively induce a pro-tumor microenvironment that permits tumor cell outgrowth into overt, clinically detectable metastatic disease. The lung is one of the most common sites of breast cancer (BC) metastasis. A chronic wound repair-related phenotype developed within the lung microenvironment during metastatic outgrowth in immunocompetent preclinical mouse models of BC. This phenotype was characterized by an increased number and activation of lung type II alveolar epithelial (AT2) cells surrounding growing metastases. Metastatic outgrowth significantly changed AT2 gene expression, resulting in a modified secretome. AT2-derived secreted factors also promote TNBC growth. AT2 secreted factors are regulated by the cAMP response element-binding protein (CREB). Targeting CREB signaling with the phosphodiesterase 4 (PDE4) inhibitor roflumilast reduced AT2-BC reciprocal interactions in vitro and metastatic outgrowth in vivo.

STATEMENT OF SIGNIFICANCEAlveolar epithelial cells are the most common cell type in the lung. Our studies demonstrate the potential for targeting metastasis-associated wound repair and lung epithelial cell activation during metastatic outgrowth with FDA-approved PDE4 inhibitors. This strategy may be an effective way to treat and manage progression of established metastatic BC.
]]></description>
<dc:creator>Christenson, J. L.</dc:creator>
<dc:creator>Spoelstra, N. S.</dc:creator>
<dc:creator>Williams, M. M.</dc:creator>
<dc:creator>O'Neill, K. I.</dc:creator>
<dc:creator>Orlicky, D. J.</dc:creator>
<dc:creator>Wagner, J. A.</dc:creator>
<dc:creator>Goodspeed, A. E.</dc:creator>
<dc:creator>Kuo, L.-W.</dc:creator>
<dc:creator>Crump, L. S.</dc:creator>
<dc:creator>Richer, J. K.</dc:creator>
<dc:date>2025-07-27</dc:date>
<dc:identifier>doi:10.1101/2025.07.23.666437</dc:identifier>
<dc:title><![CDATA[Metastasis-associated wound repair promotes reciprocal activation of the lung epithelium and breast cancer metastases during outgrowth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.27.667045v1?rss=1">
<title>
<![CDATA[
Ebola virus VP35 NNLNS motif modulates viral RNA synthesis and MIB2-mediated signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.27.667045v1?rss=1</link>
<description><![CDATA[
Ebola virus (EBOV) is a non-segmented, negative-sense virus (NNSV) with a single-stranded RNA genome. EBOV encodes for a limited number of proteins and thus depends on host factors to facilitate viral replication and pathogenesis. Of the virus-encoded proteins, multifunctional EBOV VP35 (eVP35) is necessary for host immune evasion and viral RNA synthesis. Previous proteomics studies identified an interaction between eVP35 and the host E3 ubiquitin ligase Mindbomb 2 (MIB2). Here, we show how a previously uncharacterized NNLNS motif (residues 201-205) within eVP35 serves as a binding site for MIB2. This motif is critical for eVP35-dependent inhibition of MIB2-mediated IFN induction. It is also important for EBOV RNA synthesis as MIB2 binding to eVP35 inhibited EBOV minigenome activity. Altogether, these findings highlight the importance of the eVP35 protein and the role of host factors in EBOV infection.

SIGNIFICANCE STATEMENTThe Ebola virus (EBOV) genome encodes for a limited number of proteins and depends on host factors to facilitate viral replication. Identification and characterization of host-viral interactions are needed to define infection, resolution, and to develop new therapeutics. EBOV VP35 (eVP35) is necessary for mediating host immune evasion and a cofactor for viral RNA synthesis. Here we characterized an interaction between eVP35 and MIB2. We show that the 201NNLNS205 motif in eVP35 is necessary and sufficient for MIB2 binding and inhibition of MIB2-mediated IFN production. Our results also reveal how the eVP35-MIB2 interaction impacts virus infection. These results support the importance of the multifunctional eVP35 to EBOV infection and highlight the significance of host proteins, including E3 ligases, during viral infection.
]]></description>
<dc:creator>Uwase, G.</dc:creator>
<dc:creator>Luthra, P.</dc:creator>
<dc:creator>Vogel, O. A.</dc:creator>
<dc:creator>Batra, J.</dc:creator>
<dc:creator>LaRosa, B. A.</dc:creator>
<dc:creator>Sheehan, K. C. F.</dc:creator>
<dc:creator>Khatavkar, O.</dc:creator>
<dc:creator>Payton, J.</dc:creator>
<dc:creator>Davey, R. A.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Basler, C. F.</dc:creator>
<dc:creator>Leung, D. W.</dc:creator>
<dc:creator>Amarasinghe, G. K.</dc:creator>
<dc:date>2025-07-27</dc:date>
<dc:identifier>doi:10.1101/2025.07.27.667045</dc:identifier>
<dc:title><![CDATA[Ebola virus VP35 NNLNS motif modulates viral RNA synthesis and MIB2-mediated signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.23.666478v1?rss=1">
<title>
<![CDATA[
AlignmentFold and AlignmentPartition: Improving the align-then-fold approach for RNA secondary structure prediction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.23.666478v1?rss=1</link>
<description><![CDATA[
The combination of free energy minimization with sequence covariation analysis is a popular approach for predicting conserved secondary structure using a fixed alignment of homologous RNA sequences. In this work we revisit this approach by developing the new AlignmentFold and AlignmentPartition programs. AlignmentFold and AlignmentPartition predict consensus minimum free energy structures and base pairing probabilities, respectively. We determined new nearest neighbor thermodynamic parameters for gaps and non-canonical base pairs that align with base pairs. These frequently occur for structure prediction with a fixed input alignment. We assessed the impact of alignment prediction method and size on the prediction accuracy. We also assessed the improvement in prediction accuracy conferred by sequence covariation analysis. In contrast to previous solutions, we do not include covariation in the scoring of structure quality and achieve prediction accuracy as good as other tools. AlignmentFold and AlignmentPartition are freely available as part of the RNAstructure software package at https://rna.urmc.rochester.edu/RNAstructure.html.
]]></description>
<dc:creator>Mittal, A.</dc:creator>
<dc:creator>Mathews, D. H.</dc:creator>
<dc:date>2025-07-28</dc:date>
<dc:identifier>doi:10.1101/2025.07.23.666478</dc:identifier>
<dc:title><![CDATA[AlignmentFold and AlignmentPartition: Improving the align-then-fold approach for RNA secondary structure prediction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.28.667310v1?rss=1">
<title>
<![CDATA[
Cell Cycle Arrest of a "Zippering" Epithelial Cell Cluster Shapes the Face and is Disrupted in Craniofacial Disorders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.28.667310v1?rss=1</link>
<description><![CDATA[
Facial features identify individuals, but the mechanisms shaping the human face remain elusive. Orofacial clefting (OFC), the most common craniofacial abnormality, results from failed fusion of the facial prominences that is in part caused by persistence of the cephalic epithelium. Here we uncover the identity, behaviors, and molecular blueprints of a novel craniofacial epithelial population, the Zippering Lambda (ZL), which mediates prominence fusion and is characterized by cell cycle arrest in mouse and human embryos. Remarkably, cell cycle is unleashed in the ZL of Pbx1/2 and p63 mutant mice with OFC. Intersection of ZL-enriched genes with human OFC whole-genome sequencing datasets identifies ZFHX3 variants in affected individuals and cephalic epithelial Zfhx3 deletion causes murine OFC. ZFHX3 and PBX1 genetically interact and synergistically regulate cell cycle inhibitor genes within a complex in embryonic faces. Collectively, we deconstruct new mechanisms that pattern the face, connecting cell cycle arrest to developmental tissue fusion.
]]></description>
<dc:creator>Qu, T.</dc:creator>
<dc:creator>Chacon, B. H.</dc:creator>
<dc:creator>Faure, L.</dc:creator>
<dc:creator>Losa, M.</dc:creator>
<dc:creator>Hernandez-Martinez, R.</dc:creator>
<dc:creator>Robinson, K.</dc:creator>
<dc:creator>Jones, A.</dc:creator>
<dc:creator>Lisgo, S.</dc:creator>
<dc:creator>Risolino, M.</dc:creator>
<dc:creator>De Anda, J.</dc:creator>
<dc:creator>Panagiotakos, G.</dc:creator>
<dc:creator>Leslie-Clarkson, E. J.</dc:creator>
<dc:creator>Adameyko, I.</dc:creator>
<dc:creator>Selleri, L.</dc:creator>
<dc:date>2025-07-30</dc:date>
<dc:identifier>doi:10.1101/2025.07.28.667310</dc:identifier>
<dc:title><![CDATA[Cell Cycle Arrest of a "Zippering" Epithelial Cell Cluster Shapes the Face and is Disrupted in Craniofacial Disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.25.666876v1?rss=1">
<title>
<![CDATA[
Aging-Associated Autoimmunity in Genetically Diverse UM-HET3 Mice Shows a Female Sex Bias 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.25.666876v1?rss=1</link>
<description><![CDATA[
C57BL/6 (B6) mice, often considered a non-autoimmune control strain, spontaneously develop autoantibodies and lymphocytic infiltration in the salivary glands (SG) with aging. However, as an inbred strain, B6 mice have a limited genetic background and do not fully represent a genetically diverse population. To assess whether genetic diversity influences the development of age-related autoimmunity, we studied UM-HET3 mice, a four-way cross that is commonly used in aging research. By 14-20 months of age, females showed significantly higher frequencies and endpoint titers of anti-nuclear antibodies. Older females also exhibited increased levels of splenic atypical/age-associated B cells and follicular helper T cells, populations associated with the production of autoantibodies. Similar immune cell changes were observed in the SG, with some female mice developing organized lymphocytic foci consisting of T and B cells. Our findings demonstrate that UM-HET3 mice naturally develop systemic autoimmunity and sialadenitis with age, with a clear female bias. Since female UM-HET3 mice have a longer median lifespan than males, these autoimmune responses may reflect benign autoimmunity, representing a heightened immune response associated with aging.
]]></description>
<dc:creator>Guzniczak, N.</dc:creator>
<dc:creator>Makuch, M.</dc:creator>
<dc:creator>Tillett, M.</dc:creator>
<dc:creator>Bagavant, H.</dc:creator>
<dc:creator>Deshmukh, U. S.</dc:creator>
<dc:date>2025-07-31</dc:date>
<dc:identifier>doi:10.1101/2025.07.25.666876</dc:identifier>
<dc:title><![CDATA[Aging-Associated Autoimmunity in Genetically Diverse UM-HET3 Mice Shows a Female Sex Bias]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.28.667211v1?rss=1">
<title>
<![CDATA[
A stem cell-based platform for functional analysis of genetic variants in lung disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.28.667211v1?rss=1</link>
<description><![CDATA[
Advances in genetic and transcriptomic technologies have identified large numbers of genes and variants of potential importance to human disease. Determining the function of these genes and variants is a critical bottleneck in understanding disease etiology. Variants of uncertain significance (VUS) are highly prevalent in our genomes, but our ability to identify them significantly outpaces our ability to determine their molecular and clinical consequences. We developed a genetically tractable induced pluripotent stem cell (iPSC) based platform to investigate gene variant pathogenicity in lung disease, using primary ciliary dyskinesia (PCD) as a model. We identified an individual with a clinical diagnosis of PCD and a VUS in the gene Multiciliate differentiation and DNA synthesis associated cell cycle protein (MCIDAS). Through gene-editing of iPSC-derived airway basal stem cells (iBCs), we precisely defined the molecular and cellular pathogenicity of the variant providing a successful application of the iPSC system to diagnose a lung disease.
]]></description>
<dc:creator>Wallman, D. J.</dc:creator>
<dc:creator>Homma, S. T.</dc:creator>
<dc:creator>Finton, M.</dc:creator>
<dc:creator>Beermann, M.</dc:creator>
<dc:creator>Jacob, A.</dc:creator>
<dc:creator>Choksi, S.</dc:creator>
<dc:creator>Pletcher, S. D.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Bawa, P. S.</dc:creator>
<dc:creator>Reiter, J. F.</dc:creator>
<dc:creator>Horani, A.</dc:creator>
<dc:creator>Brody, S. L.</dc:creator>
<dc:creator>Berical, A.</dc:creator>
<dc:creator>Hawkins, F. J.</dc:creator>
<dc:date>2025-07-31</dc:date>
<dc:identifier>doi:10.1101/2025.07.28.667211</dc:identifier>
<dc:title><![CDATA[A stem cell-based platform for functional analysis of genetic variants in lung disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.30.667769v1?rss=1">
<title>
<![CDATA[
Modulation of the 14-3-3σ/C-RAF autoinhibited complexby molecular glues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.30.667769v1?rss=1</link>
<description><![CDATA[
Molecular glues, compounds that bind cooperatively at protein-protein interfaces are revolutionizing chemical biology and drug discovery, allowing the modulation of traditional "undruggable" targets. Here, we focus on the native protein-protein interaction (PPI) of C-RAF, a key component of the MAPK signaling pathway, with the scaffolding protein 14-3-3. Although extensive drug discovery efforts have focused on the MAPK pathway, its central role in oncology and developmental disorders (RASopathies), still requires alternative approaches, moving beyond direct kinase inhibition. Indeed, stabilization of native PPIs is a relatively unexplored territory in this pathway. The function of C-RAF is regulated on multiple levels including dimerization, phosphorylation and complex formation with the hub protein 14-3-3. 14-3-3 prevents C-RAF activation by molecular recognition and binding at the phospho-serine 259. We used a fragment-merging approach to design a molecular glue scaffold that would bind to the composite surface of the 14-3-3/C-RAF "auto"inhibited complex. The synthesized molecular glues stabilized the 14-3-3/C-RAF complex up to 300-fold in biophysical assays; their glue-based mechanism of action was confirmed with several crystal structures of ternary complexes. Selectivity among the other RAF isoforms and other RAF phosphorylation sites was evaluated with biophysical assays. The best compounds showed excellent selectivity among a broad panel of 80 14-3-3 clients. Validation in cell assays showed on-target engagement, enhanced phosphorylation levels of the C-RAF pS259 site, reduced RAF dimerization and reduced ERK phosphorylation. Overall, this approach enables chemical biology studies on a C-RAF site that is intrinsically disordered prior to 14-3-3 binding and has not been targeted previously. These molecular glues will be useful as chemical probes and starting points for further drug discovery efforts to elucidate the effect of native PPI stabilization in the MAPK pathway with applications in oncology and RASopathies.
]]></description>
<dc:creator>Konstantinidou, M.</dc:creator>
<dc:creator>Vickery, H. R.</dc:creator>
<dc:creator>Pennings, M. A. M.</dc:creator>
<dc:creator>Virta, J. M.</dc:creator>
<dc:creator>Visser, E. J.</dc:creator>
<dc:creator>Bannier, S. D.</dc:creator>
<dc:creator>Srikanth, M.</dc:creator>
<dc:creator>Cismoski, S. Z.</dc:creator>
<dc:creator>Young, L. C.</dc:creator>
<dc:creator>van den Oetelaar, M. C. M.</dc:creator>
<dc:creator>McCromick, F.</dc:creator>
<dc:creator>Ottmann, C.</dc:creator>
<dc:creator>Brunsveld, L.</dc:creator>
<dc:creator>Arkin, M. R.</dc:creator>
<dc:date>2025-07-31</dc:date>
<dc:identifier>doi:10.1101/2025.07.30.667769</dc:identifier>
<dc:title><![CDATA[Modulation of the 14-3-3σ/C-RAF autoinhibited complexby molecular glues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.29.667331v1?rss=1">
<title>
<![CDATA[
BBSome-Mediated Clearance of Ubiquitinated IMPG2 Defines a Constitutive Ciliary Retrieval Pathway in Photoreceptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.29.667331v1?rss=1</link>
<description><![CDATA[
The BBSome mediates the retrieval of ubiquitinated membrane proteins from cilia, but its physiological cargoes in photoreceptors remain largely unidentified. Here, we find that K63-linked ubiquitin (UbK63) chains accumulate in the outer segment (OS, equivalent of cilia) of Bbs4-/- photoreceptors from the onset of OS formation. Through quantitative profiling of the UbK63-associated OS proteome, we identify the transmembrane fragment of interphotoreceptor matrix proteoglycan 2 (IMPG2m) as a principal cargo of the BBSome. In Bbs4-/- mice, ubiquitinated IMPG2m aberrantly accumulates in OSs, and disruption of IMPG2m ubiquitination impairs its retrieval and clearance. Because full-length IMPG2 traffics to the OS to deliver its extracellular domain to the matrix, our data support a model in which IMPG2m undergoes constitutive cycling between the inner and outer segments. These findings redefine the BBSomes role in photoreceptors from quality control to constitutive membrane protein turnover.
]]></description>
<dc:creator>Das, T.</dc:creator>
<dc:creator>Bradshaw, G. A.</dc:creator>
<dc:creator>Hyer, J.</dc:creator>
<dc:creator>Masek, M.</dc:creator>
<dc:creator>Kuo, Y.-M.</dc:creator>
<dc:creator>Bachmann-Gagescu, R.</dc:creator>
<dc:creator>Kalocsay, M.</dc:creator>
<dc:creator>Nachury, M. V.</dc:creator>
<dc:date>2025-08-01</dc:date>
<dc:identifier>doi:10.1101/2025.07.29.667331</dc:identifier>
<dc:title><![CDATA[BBSome-Mediated Clearance of Ubiquitinated IMPG2 Defines a Constitutive Ciliary Retrieval Pathway in Photoreceptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.29.667471v1?rss=1">
<title>
<![CDATA[
Alanine Catabolism as a Targetable Vulnerability for MYC-Driven Liver Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.29.667471v1?rss=1</link>
<description><![CDATA[
Liver cancer is a leading cause of cancer-related death world-wide in part due to the shortage of effective therapies, and MYC overexpression defines an aggressive and especially difficult to treat subset of patients. Given MYCs ability to reprogram cancer cell metabolism, and the livers role as a coordinator of systemic metabolism, we hypothesized that MYC induces metabolic dependencies that could be targeted to attenuate liver tumor growth. We discovered that MYC-driven liver cancers catabolize alanine in a GPT2-dependent manner to sustain their growth. GPT2 is the predominant alanine-catabolizing enzyme expressed in MYC-driven liver tumors and genetic ablation of GPT2 limited MYC-driven liver tumorigenesis. In vivo isotope tracing studies uncovered a role for alanine as a substrate for a repertoire of pathways including the tricarboxylic acid cycle, nucleotide production, and amino acid synthesis. Treating transgenic MYC-driven liver tumor mouse models with L-Cycloserine, a compound that inhibits GPT2, was sufficient to diminish the frequency of mouse tumor formation and attenuate growth of established human liver tumors. Thus, we identify a new targetable metabolic dependency that MYC-driven liver tumors usurp to ensure their survival.
]]></description>
<dc:creator>Montoya, T.</dc:creator>
<dc:creator>Lee, J. V.</dc:creator>
<dc:creator>Qiu, L.</dc:creator>
<dc:creator>Krall, A. V.</dc:creator>
<dc:creator>Matulionis, N.</dc:creator>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>Finck, B.</dc:creator>
<dc:creator>Kelley, R. K.</dc:creator>
<dc:creator>Christofk, H.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:date>2025-08-01</dc:date>
<dc:identifier>doi:10.1101/2025.07.29.667471</dc:identifier>
<dc:title><![CDATA[Alanine Catabolism as a Targetable Vulnerability for MYC-Driven Liver Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.29.667509v1?rss=1">
<title>
<![CDATA[
The Lipid Hydrolase ABHD6 is a Therapeutic Target in Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD)-Related Hepatocellular Carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.29.667509v1?rss=1</link>
<description><![CDATA[
Primary liver cancer accounts for approximately 700,000 deaths worldwide annually ranking third in cancer-related mortality, with hepatocellular carcinoma (HCC) comprising the majority of these tumors. Metabolic dysfunction-associated steatotic liver disease (MASLD) is currently a leading cause of HCC in the United States. We previously identified the lipid hydrolase alpha/beta hydrolase domain 6 (ABHD6) as a key mediator of the development of metabolic syndrome and intimately involved in cell signaling, making it a prime target for investigation in MASLD-related HCC. ABHD6 displays higher expression within HCC tumor cores when compared to adjacent non-tumor liver tissue in human subjects. Using an in vivo antisense oligonucleotide (ASO)-driven knockdown approach, we have shown the inhibition of ABHD6 prevents the development and progression of HCC in an obesity/MASLD-driven mouse model. Additionally, a xenograft model using the human Huh7 cell line displayed reduced tumor engraftment and growth with ABHD6 genetic deletion and small molecule inhibition. ABHD6 knockout cells demonstrated increased levels of bis(monoacylglycerol)phosphates (BMPs), lipids relevant to high fat diet-induced lysosomal dysfunction, and knockout cells also demonstrated altered autophagy and lysosomal activity using in vitro model of saturated fatty acid-induced lipotoxicity. These studies reveal novel lipid signaling mechanisms by which MASLD progresses towards HCC and provide support for ABHD6 as a therapeutic target in HCC.

SignificanceWe have identified that alpha/beta hydrolase domain 6 (ABHD6) plays a role in lysosomal membrane lipid remodeling pathways that are relevant in obesity/MASLD-driven HCC. Inhibitors targeting ABHD6 reorganize lysosomal lipid homeostasis to improve outcomes in HCC.
]]></description>
<dc:creator>Orabi, D.</dc:creator>
<dc:creator>Massey, W.</dc:creator>
<dc:creator>Fung, K.</dc:creator>
<dc:creator>Varadharajan, V.</dc:creator>
<dc:creator>Ramachandiran, I.</dc:creator>
<dc:creator>Banerjee, R.</dc:creator>
<dc:creator>Silver, D. J.</dc:creator>
<dc:creator>Osborn, L. J.</dc:creator>
<dc:creator>Brown, A. L.</dc:creator>
<dc:creator>Marshall, S.</dc:creator>
<dc:creator>Ferguson, D.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Schugar, R. C.</dc:creator>
<dc:creator>Finney, C.</dc:creator>
<dc:creator>Neumann, C.</dc:creator>
<dc:creator>Burrows, A. C.</dc:creator>
<dc:creator>Horak, A. J.</dc:creator>
<dc:creator>Pathak, P.</dc:creator>
<dc:creator>Helsley, R. N.</dc:creator>
<dc:creator>Bulfon, D.</dc:creator>
<dc:creator>Zimmermann, R.</dc:creator>
<dc:creator>Leung, Y. H.</dc:creator>
<dc:creator>Madiraju, S. R. M.</dc:creator>
<dc:creator>Prentki, M.</dc:creator>
<dc:creator>Lee, R. G.</dc:creator>
<dc:creator>Mullick, A. E.</dc:creator>
<dc:creator>Zergeeva, O.</dc:creator>
<dc:creator>Dasarathy, S.</dc:creator>
<dc:creator>Lee, Z.</dc:creator>
<dc:creator>Allende, D. S.</dc:creator>
<dc:creator>Aucejo, F.</dc:creator>
<dc:creator>Lathia, J. D.</dc:creator>
<dc:creator>Brown, J. M.</dc:creator>
<dc:date>2025-08-02</dc:date>
<dc:identifier>doi:10.1101/2025.07.29.667509</dc:identifier>
<dc:title><![CDATA[The Lipid Hydrolase ABHD6 is a Therapeutic Target in Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD)-Related Hepatocellular Carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.30.667738v1?rss=1">
<title>
<![CDATA[
A novel locus associated with decreased susceptibility of Plasmodium falciparum to lumefantrine and dihydroartemisinin has emerged and spread in Uganda 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.30.667738v1?rss=1</link>
<description><![CDATA[
Malaria control in Uganda is threatened by the emergence of artemisinin partial resistance (ART-R) and decreasing lumefantrine susceptibility. To identify loci contributing to decreased drug susceptibility, we assessed signatures of selection in Ugandan whole genome Plasmodium falciparum sequences. Extended shared haplotypes were seen for the ART-R associated Kelch13 (K13) C469Y and A675V mutations, but the strongest signal of recent selection was centered on a segment of chromosome 7 encoding the phosphoinositide-binding protein gene (px1, PF3D7_0720700). A haplotype, represented by three PX1 SNPs (L1222P, M1701I and D1705N) and two indels (designated PIN), was first seen in 2008 and rapidly increased, reaching prevalence >50% in northern Uganda by 2016 and eastern Uganda by 2023. PIN-carrying parasites showed significantly decreased ex vivo susceptibilities, measured as IC50s, to lumefantrine, mefloquine and dihydroartemisinin, either with or without coincident K13 mutations. Thus, PX1 polymorphisms may impact on the susceptibilities of African malaria parasites to key drugs.
]]></description>
<dc:creator>Niare, K.</dc:creator>
<dc:creator>Tafesse, B.</dc:creator>
<dc:creator>Treat, M.</dc:creator>
<dc:creator>Sadler, J.</dc:creator>
<dc:creator>Okitwi, M.</dc:creator>
<dc:creator>Orena, S.</dc:creator>
<dc:creator>Asua, V.</dc:creator>
<dc:creator>Kreutzfeld, O.</dc:creator>
<dc:creator>Legac, J.</dc:creator>
<dc:creator>Samuel, N. L.</dc:creator>
<dc:creator>Yeka, A.</dc:creator>
<dc:creator>Rosenthal, P. J.</dc:creator>
<dc:creator>Juliano, J. J.</dc:creator>
<dc:creator>Bailey, J. A.</dc:creator>
<dc:creator>Conrad, M. D.</dc:creator>
<dc:date>2025-08-02</dc:date>
<dc:identifier>doi:10.1101/2025.07.30.667738</dc:identifier>
<dc:title><![CDATA[A novel locus associated with decreased susceptibility of Plasmodium falciparum to lumefantrine and dihydroartemisinin has emerged and spread in Uganda]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.01.668069v1?rss=1">
<title>
<![CDATA[
The Rpd3 histone deacetylase is critical for temperature-mediated morphogenesis and virulence in the human fungal pathogen Histoplasma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.01.668069v1?rss=1</link>
<description><![CDATA[
Adaptive responses to environmental stimuli are integral to the survival and virulence of microbial pathogens. The thermally dimorphic human fungal pathogen Histoplasma senses temperature to transition between a mold form in soil and a pathogenic yeast in mammalian hosts. The contributions of chromatin-modifying enzymes to the ability of Histoplasma to appropriately respond to temperature have never been explored. Through chemical inhibition and genetics, we determined that the class I histone deacetylase (HDAC) RPD3 is required for normal Histoplasma yeast morphology at 37 {degrees}C. Rpd3 regulated the expression of key morphology-specific genes, including critical virulence factors and transcription factors (TFs), was required for normal DNA-binding activity of yeast-promoting TFs, and influenced histone acetylation levels at the loci of putative pro-filamentation TFs. Furthermore, Rpd3 was required for virulence in a macrophage model of infection. Taken together, Rpd3 is a critical regulatory component that both activates the pathogenesis program and represses the filamentation program to enable thermal dimorphism in Histoplasma. This work uncovers the crucial role that chromatin regulation plays in temperature response of this ubiquitous pathogen.
]]></description>
<dc:creator>Ali, N. B.</dc:creator>
<dc:creator>Voorhies, M.</dc:creator>
<dc:creator>Rodriguez, R. A.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:date>2025-08-04</dc:date>
<dc:identifier>doi:10.1101/2025.08.01.668069</dc:identifier>
<dc:title><![CDATA[The Rpd3 histone deacetylase is critical for temperature-mediated morphogenesis and virulence in the human fungal pathogen Histoplasma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.04.668434v1?rss=1">
<title>
<![CDATA[
Systemic translocation of S. aureus Drives Anti-CD4 Autoimmunity in Treated HIV Infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.04.668434v1?rss=1</link>
<description><![CDATA[
BackgroundIn 2017, our group first demonstrated that autoimmunity contributes to HIV pathogenesis, even without autoimmune disease. This concept is now broadly recognized, exemplified by the role of autoimmunity in severe COVID-19. In people with HIV (PWH) on suppressive ART, anti-CD4 autoantibodies may impair CD4+ T cell recovery, though the mechanisms driving their production remain unclear. Building on evidence from our group and others that Staphylococcus aureus and its peptidoglycan (PGN) promote autoimmunity, we investigated their contribution to anti-CD4 IgG in HIV.

MethodsPlasma from 32 ART-naive PWH, 53 ART-treated PWH, and 32 HIV-negative controls was analyzed for IgG autoantibodies and markers of S. aureus translocation using protein array, ELISA, and microarray. EcoHIV mice were injected intraperitoneally with saline, S. aureus PGN, or Bacillus subtilis PGN. PGN structures were compared by mass spectrometry.

ResultsAmong 87 autoantibodies, 40% were elevated in ART-naive PWH and largely normalized by ART; however, anti-CD4 IgGs remained elevated in PWH on ART. Anti-CD4 IgG levels inversely correlated with CD4+ T cell counts in ART-treated PWH and positively with S. aureus translocation. In mice, S. aureus PGN induced anti-CD4 IgGs, reduced gut CD4+ T cells, and promoted surface IgG binding and apoptosis in CD4+ T cells.

ConclusionS. aureus and its PGN translocation may drive anti-CD4 autoimmunity and hinder immune recovery in PWH on suppressive ART, highlighting S. aureus colonization as a therapeutic target and supporting the development of competitive probiotic interventions such as Bacillus subtilis.
]]></description>
<dc:creator>Jiang, W.</dc:creator>
<dc:creator>Da, C.</dc:creator>
<dc:creator>Luo, Z.</dc:creator>
<dc:creator>Ning, W.</dc:creator>
<dc:creator>Heath, S. L.</dc:creator>
<dc:creator>Gisslen, M.</dc:creator>
<dc:creator>Price, R. W.</dc:creator>
<dc:creator>Adekunle, R.</dc:creator>
<dc:creator>Salman, T.</dc:creator>
<dc:creator>Johnson, D.</dc:creator>
<dc:creator>McKinnon, J. E.</dc:creator>
<dc:creator>Hu, W.</dc:creator>
<dc:creator>Ndhlovu, L.</dc:creator>
<dc:date>2025-08-04</dc:date>
<dc:identifier>doi:10.1101/2025.08.04.668434</dc:identifier>
<dc:title><![CDATA[Systemic translocation of S. aureus Drives Anti-CD4 Autoimmunity in Treated HIV Infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.04.668450v1?rss=1">
<title>
<![CDATA[
Targeting the COP9 signalosome overcomes platinum resistance in ovarian cancer through two distinct genome stability mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.04.668450v1?rss=1</link>
<description><![CDATA[
Tubo-ovarian high-grade serous carcinoma (HGSC) is a leading cause of gynecologic cancer mortality, largely due to the emergence of platinum resistance, which serves as the mainstay of chemotherapy. Here, we identify COPS5 as a therapeutic target and use an available small molecule inhibitor to overcome platinum resistance. A genetic screen for platinum-induced DNA damage in a platinum resistant ovarian cancer model identified COPS5 and COPS6, two components of the COP9 signalosome. Consistently, high COPS5 expression correlated with poor clinical outcomes in patients with HGSC. In both in vitro and in vivo experiments, COPS5 depletion sensitized ovarian cancer cells to carboplatin. A small molecule COPS5 inhibitor, CSN5i-3, synergized with carboplatin in homologous recombination-deficient and -proficient cells. This combination was also effective in xenografts and in a syngeneic mouse model of carboplatin-resistant HGSC. Importantly, we demonstrate that CSN5i-3 is selective for cancer cells, with patient-derived HGSC cells exhibiting up to 50-fold greater sensitivity to CSN5i-3 than benign cells. Finally, we show that genetic or small molecule inhibition of COPS5 impaired both nucleotide excision repair (NER) and interstrand crosslink (ICL) repair, leading to increased DNA platinum adducts. Mechanistically, this was due to increased ubiquitination and degradation of DNA-specific DNA binding protein 1 (DDB1) and other key NER and ICL repair proteins, consistent with the role of COPS5 in the regulation of these factors. Our findings highlight the importance of NER and ICL regulation in chemotherapy response and indicate that targeting COPS5 can enhance the efficacy of platinum-based chemotherapy in HGSC.

One Sentence SummaryCOPS5 depletion or inhibition using a small molecule COPS5 inhibitor CSN5i-3 sensitizes high-grade serous carcinoma to platinum chemotherapy through downregulation of nucleotide excision repair and interstrand crosslink repair.
]]></description>
<dc:creator>Lomonosova, E.</dc:creator>
<dc:creator>Loeb, M.</dc:creator>
<dc:creator>Rodriguez, K.</dc:creator>
<dc:creator>van Biljon, L. N.</dc:creator>
<dc:creator>Brill, J.</dc:creator>
<dc:creator>Schab, A.</dc:creator>
<dc:creator>Minett, M.</dc:creator>
<dc:creator>Barron, J.</dc:creator>
<dc:creator>Bittner, M.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Drexler, R.</dc:creator>
<dc:creator>Sandoval, C.</dc:creator>
<dc:creator>Oplt, A.</dc:creator>
<dc:creator>Gallup, E.</dc:creator>
<dc:creator>Thuthika, P. K.</dc:creator>
<dc:creator>Sadeghipour, N.</dc:creator>
<dc:creator>Wu, S.</dc:creator>
<dc:creator>Oberley, M. J.</dc:creator>
<dc:creator>Sanders, B.</dc:creator>
<dc:creator>Kuroki, L.</dc:creator>
<dc:creator>McCourt, C.</dc:creator>
<dc:creator>Hagemann, A. R.</dc:creator>
<dc:creator>Thaker, P.</dc:creator>
<dc:creator>Mutch, D.</dc:creator>
<dc:creator>Powell, M.</dc:creator>
<dc:creator>Hagemann, I. S.</dc:creator>
<dc:creator>Ilagan, M.</dc:creator>
<dc:creator>Fuh, K.</dc:creator>
<dc:creator>Verma, P.</dc:creator>
<dc:creator>Scharer, O.</dc:creator>
<dc:creator>Mosammaparast, N.</dc:creator>
<dc:creator>Khabele, D.</dc:creator>
<dc:creator>Mullen, M. M.</dc:creator>
<dc:date>2025-08-06</dc:date>
<dc:identifier>doi:10.1101/2025.08.04.668450</dc:identifier>
<dc:title><![CDATA[Targeting the COP9 signalosome overcomes platinum resistance in ovarian cancer through two distinct genome stability mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.04.668485v1?rss=1">
<title>
<![CDATA[
Reversible therapeutic resistance in EGFR mutant lung cancer caused by RB1 loss induced lineage plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.04.668485v1?rss=1</link>
<description><![CDATA[
Treatment with EGFR tyrosine kinase inhibitors (EGFR TKI) is standard of care for patients with lung cancer initiated by activating mutations in the EGFR gene. While EGFR TKI treatment is effective, virtually all patients progress on therapy. Loss of function mutations in the RB1 tumor suppressor gene are associated with poor EGFR TKI outcomes, but underlying mechanisms remain unclear. Analysis of genetically engineered experimental models and human clinical specimens described here demonstrates RB1 loss has minimal impact on initial cell cycle responses to EGFR TKI but markedly accelerates acquired resistance over time. RB1 loss promotes transcriptional plasticity that enables rare cells to spontaneously assume reversible, resistance-conferring lineage states. These findings highlight the importance of non-genetic adaptations in driving acquired EGFR TKI resistance, advance our understanding of how RB1 impacts cancer, and identify opportunities to improve EGFR TKI treatment outcomes by targeting reversible epigenetic resistance mechanisms.
]]></description>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Chinnam, M.</dc:creator>
<dc:creator>Balk, B.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Shaurova, T.</dc:creator>
<dc:creator>Alam, S.</dc:creator>
<dc:creator>Pokharel, S.</dc:creator>
<dc:creator>Singh, P.</dc:creator>
<dc:creator>Jacobi, J.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Cortes Gomez, E.</dc:creator>
<dc:creator>Habitzruther, M.</dc:creator>
<dc:creator>Seshadri, M.</dc:creator>
<dc:creator>Atanassov, B.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Benevolenskaya, E.</dc:creator>
<dc:creator>Blakely, C. M.</dc:creator>
<dc:creator>Hershberger, P.</dc:creator>
<dc:creator>Long, M.</dc:creator>
<dc:creator>Bivona, T.</dc:creator>
<dc:creator>Goodrich, D. W.</dc:creator>
<dc:date>2025-08-06</dc:date>
<dc:identifier>doi:10.1101/2025.08.04.668485</dc:identifier>
<dc:title><![CDATA[Reversible therapeutic resistance in EGFR mutant lung cancer caused by RB1 loss induced lineage plasticity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.05.668786v1?rss=1">
<title>
<![CDATA[
Evidence for the major role of PH4EFB in the prolyl 4-hydroxylation of Drosophila collagen IV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.05.668786v1?rss=1</link>
<description><![CDATA[
Collagens are fundamental components of extracellular matrices, requiring precise intracellular post-translational modifications for proper function. Among the modifications, prolyl 4-hydroxylation is critical to stabilise the collagen triple helix. In humans, this reaction is mediated by collagen prolyl 4-hydroxylases (P4Hs). While humans possess three genes encoding these enzymes (P4Hs), Drosophila melanogaster harbour at least 26 candidates for collagen P4Hs despite its simple genome, and it is poorly understood which of them are actually working on collagen in the fly. In this study, we addressed this question by carrying out thorough bioinformatic and biochemical analyses. We demonstrate that among the 26 potential collagen P4Hs, PH4EFB shares the highest homology with vertebrate collagen P4Hs. Furthermore, while collagen P4Hs and their substrates must exist in the same cells, our transcriptomic analyses at the tissue and single cell levels showed a global co-expression of PH4EFB but not the other P4H-related genes with the collagen IV genes. Moreover, expression of PH4EFB during embryogenesis was found to precede that of collagen IV, presumably enabling efficient collagen modification by PH4EFB. Finally, biochemical assays confirm that PH4EFB binds collagen, supporting its direct role in collagen IV modification. Collectively, we identify PH4EFB as the primary and potentially constitutive prolyl 4-hydroxylase responsible for collagen IV biosynthesis in Drosophila. Our findings highlight the remarkably simple nature of Drosophila collagen IV biosynthesis, which may serve as a blueprint for defining the minimal requirements for collagen engineering.
]]></description>
<dc:creator>Ishikawa, Y.</dc:creator>
<dc:creator>Toups, M. A.</dc:creator>
<dc:creator>Elkrewi, M.</dc:creator>
<dc:creator>Zajac, A. L.</dc:creator>
<dc:creator>Horne-Badovinac, S.</dc:creator>
<dc:creator>Matsubayashi, Y.</dc:creator>
<dc:date>2025-08-06</dc:date>
<dc:identifier>doi:10.1101/2025.08.05.668786</dc:identifier>
<dc:title><![CDATA[Evidence for the major role of PH4EFB in the prolyl 4-hydroxylation of Drosophila collagen IV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.05.664214v1?rss=1">
<title>
<![CDATA[
The Placental Transcriptome Serves as a Mechanistic Link between Prenatal Phthalate Exposure and Placental Efficiency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.05.664214v1?rss=1</link>
<description><![CDATA[
Prenatal exposure to phthalates, pervasive endocrine-disrupting chemicals, has been linked to child health outcomes, including prematurity and low birthweight. Placental transcriptomics data can reveal mechanisms by which environmental toxicants alter placental and fetal growth. This study aims to investigate the placental transcriptome as a mediator between prenatal maternal urinary phthalate metabolites and placental efficiency. We identified significant associations between maternal urinary concentrations of two phthalate metabolites and the placental transcriptome (132 genes and 27 gene modules). 7 genes and 3 gene modules exhibited significant consistent mediation of the relationship between phthalates and placental efficiency measures. These genes were involved in syncytialization, metabolism, DNA damage and cellular senescence, and steroid biosynthesis--processes essential to fetal growth and development because of the placentas role in nutrient supply, hormone production, and detoxification. These findings suggest a key mediating role of the placental transcriptome in toxicological mechanisms by which phthalates may disrupt fetal growth.

TeaserPlacental gene expression mediates the relationship between prenatal phthalate exposure and fetal growth measures.
]]></description>
<dc:creator>Parenti, M.</dc:creator>
<dc:creator>Lapehn, S.</dc:creator>
<dc:creator>MacDonald, J.</dc:creator>
<dc:creator>Bammler, T.</dc:creator>
<dc:creator>Szpiro, A.</dc:creator>
<dc:creator>Hazlehurst, M.</dc:creator>
<dc:creator>Day, D. B.</dc:creator>
<dc:creator>Thoreson, C.</dc:creator>
<dc:creator>Kannan, K.</dc:creator>
<dc:creator>Bush, N. R.</dc:creator>
<dc:creator>LeWinn, K. Z.</dc:creator>
<dc:creator>Zhao, Q.</dc:creator>
<dc:creator>Sathyanarayana, S.</dc:creator>
<dc:creator>Paquette, A. G.</dc:creator>
<dc:date>2025-08-07</dc:date>
<dc:identifier>doi:10.1101/2025.08.05.664214</dc:identifier>
<dc:title><![CDATA[The Placental Transcriptome Serves as a Mechanistic Link between Prenatal Phthalate Exposure and Placental Efficiency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.05.668605v1?rss=1">
<title>
<![CDATA[
Defined human tri-lineage brain microtissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.05.668605v1?rss=1</link>
<description><![CDATA[
Microglia are the immune cells of the central nervous system and are thought to be key players in both physiological and disease conditions. Several microglial features are poorly conserved between mice and human, such as the function of the neurodegeneration-associated immune receptor Trem2. Induced pluripotent stem cell (iPSC)-derived microglia offer a powerful opportunity to generate and study human microglia. However, human iPSC-derived microglia often exhibit activated phenotypes in vitro, and assessing their impact on other brain cell types remains challenging due to limitations in current co-culture systems. Here, we developed fully defined brain microtissues, composed of human iPSC-derived neurons, astrocytes, and microglia, co-cultured in 2D or 3D formats. Our microtissues are stable and self-sufficient over time, requiring no exogenous cytokines or growth factors. All three cell types exhibit morphologies characteristic of their in vivo environment and show functional properties. Co-cultured microglia develop more homeostatic phenotypes compared to microglia exposed to exogenous cytokines. Hence, these tri-cultures provide a unique approach to investigate cell-cell interactions between brain cell types. We found that astrocytes and not neurons are sufficient for microglial survival and maturation, and that astrocyte-derived M-CSF is essential for microglial survival. Single-cell and single-nucleus RNA sequencing analyses nominated a network of reciprocal communication between cell types. Brain microtissues faithfully recapitulated pathogenic -synuclein seeding and aggregation, suggesting their usefulness as human cell models to study not only normal but also pathological cell biological processes.
]]></description>
<dc:creator>Uenaka, T.</dc:creator>
<dc:creator>Jung, S.</dc:creator>
<dc:creator>Kumar, I.</dc:creator>
<dc:creator>Vodehnal, K.</dc:creator>
<dc:creator>Rastogi, M.</dc:creator>
<dc:creator>Yoo, Y.</dc:creator>
<dc:creator>Koontz, M.</dc:creator>
<dc:creator>Thome, C.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Chan, T.</dc:creator>
<dc:creator>Green, E. M.</dc:creator>
<dc:creator>Chesnov, K.</dc:creator>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Venida, A.</dc:creator>
<dc:creator>Mahul Mellier, A.-L.</dc:creator>
<dc:creator>Atkins, M.</dc:creator>
<dc:creator>Jackrel, M.</dc:creator>
<dc:creator>Skotheim, J. M.</dc:creator>
<dc:creator>Wyss-Coray, T.</dc:creator>
<dc:creator>Abu-Remaileh, M.</dc:creator>
<dc:creator>Lashuel, H. A.</dc:creator>
<dc:creator>Bassik, M. C.</dc:creator>
<dc:creator>Sudhof, T. C.</dc:creator>
<dc:creator>del Sol, A.</dc:creator>
<dc:creator>Ullian, E.</dc:creator>
<dc:creator>Wernig, M.</dc:creator>
<dc:date>2025-08-07</dc:date>
<dc:identifier>doi:10.1101/2025.08.05.668605</dc:identifier>
<dc:title><![CDATA[Defined human tri-lineage brain microtissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.07.669008v1?rss=1">
<title>
<![CDATA[
Comparative evaluation of genomic footprinting algorithms for predicting transcription factor binding sites in single-cell data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.07.669008v1?rss=1</link>
<description><![CDATA[
Transcription factors (TFs) have millions of potential binding sites across the human genome, but only a fraction are bound in a given context. Genomic footprinting aims to identify context-specific binding sites by detecting patterns in open chromatin data. While powerful, these approaches face technical challenges, especially in single-cell applications. We developed a benchmarking framework for cell-type specific footprinting and used it to evaluate the consistency, reproducibility, and equivalency of three leading methods across data quality scenarios and as a function of cell-type similarity. Peak-level read coverage emerged as the strongest predictor of stable footprints. Motivated by limited reproducibility across tools, we built an ensemble model that improved concordance with ChIP-seq. To encourage broader adoption and continued tool development, we provide practical guidelines for robust genomic footprinting in single-cell datasets and a roadmap for extracting deeper insights about how gene regulatory networks vary across cell types in complex tissues.
]]></description>
<dc:creator>Everitt, A.</dc:creator>
<dc:creator>Whalen, S.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2025-08-07</dc:date>
<dc:identifier>doi:10.1101/2025.08.07.669008</dc:identifier>
<dc:title><![CDATA[Comparative evaluation of genomic footprinting algorithms for predicting transcription factor binding sites in single-cell data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.09.669414v1?rss=1">
<title>
<![CDATA[
Circuit inhibition promotes the dynamic reorganization of prefrontal task encoding to support cognitive flexibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.09.669414v1?rss=1</link>
<description><![CDATA[
The mammalian prefrontal cortex encodes variables related to goal-directed behavior, and enables flexibility during environmental changes, making it critical to understand how the dynamic updating vs. stable maintenance of different encodings contribute to behavioral adaptation. We addressed this by comparing prefrontal encoding during successful adaptation vs. maladaptive perseveration. Specifically, we studied mutant (Dlx5/6+/-) mice, which have dysfunctional parvalbumin-expressing inhibitory interneurons and perseverate in a rule shifting task. We measured mPFC activity patterns using microendoscopic calcium imaging, then used linear classifiers and neural networks to compare representational geometries in wild-type mice and Dlx5/6+/- mutants before, during, and after benzodiazepine treatment, which persistently rescues their rule shift learning. The encoding of correct vs. incorrect trial outcomes rapidly shifts as mice successfully learn new cue-reward associations, but becomes more stable when mutant mice perseverate. We also find activity patterns that normally distinguish learning of the initial association vs. rule shift, but become diminished during perseveration. Finally, during perseveration, outdated representations are inappropriately reinstated, not just passively maintained. These results reveal prefrontal contributions to flexible behavior driven by the dynamic reorganization of abstract rule representations, rather than stable reinforcement signals.
]]></description>
<dc:creator>Johnson-Cruz, C.</dc:creator>
<dc:creator>Cho, K. K. A.</dc:creator>
<dc:creator>Sohal, V. S.</dc:creator>
<dc:date>2025-08-11</dc:date>
<dc:identifier>doi:10.1101/2025.08.09.669414</dc:identifier>
<dc:title><![CDATA[Circuit inhibition promotes the dynamic reorganization of prefrontal task encoding to support cognitive flexibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.08.668693v1?rss=1">
<title>
<![CDATA[
Genetic and Epigenetic Reprogramming of Transposable Elements Drives ecDNA-Mediated Metastatic Prostate Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.08.668693v1?rss=1</link>
<description><![CDATA[
Extrachromosomal DNAs (ecDNAs), which replicate and segregate in a non-Mendelian manner, serve as vectors for accelerated tumor evolution. By integrating chromatin accessibility, whole-genome sequencing, and Hi-C-based genome topology data from a cohort of metastatic Castration-Resistant Prostate Cancer (mCRPC) cases, we show that epigenetically activated repeat DNA, amplified in ecDNAs, drive oncogene overexpression. Specifically, we identify a subgroup of mCRPCs (20%) characterized by clusters of accessible LINE1 repeat DNA elements flanking the androgen receptor (AR) gene. These LINE1 elements are co-amplified with AR and provide binding sites for prostate-lineage transcription factors, including AR, FOXA1 and HOXB13. Accessible LINE1 elements establish novel 3D chromatin interactions with the AR gene, forging a new regulatory plexus driving AR overexpression and confers resistance to androgen signaling inhibitors. Our findings indicate how tumor evolution is driven by the convergence of genetic and epigenetic alterations on repeat DNA, activating and amplifying them to allow oncogene overexpression.

Statement of significanceWe show how tumor evolution is driven by the convergence of genetic and epigenetic alterations on repeat DNA elements, resulting in their activation as regulatory elements and co-amplification in ecDNAs with oncogenes in mCRPC.
]]></description>
<dc:creator>Mout, L.</dc:creator>
<dc:creator>Moreno-Rodriguez, T.</dc:creator>
<dc:creator>Grillo, G.</dc:creator>
<dc:creator>Nand, A.</dc:creator>
<dc:creator>Keshavarzian, T.</dc:creator>
<dc:creator>Bahl, S.</dc:creator>
<dc:creator>Kang, K.</dc:creator>
<dc:creator>Bootsma, M.</dc:creator>
<dc:creator>Minnee, E.</dc:creator>
<dc:creator>Zhou, S.</dc:creator>
<dc:creator>Burns, K. H.</dc:creator>
<dc:creator>Corey, E.</dc:creator>
<dc:creator>Nelson, P.</dc:creator>
<dc:creator>Dehm, S. M.</dc:creator>
<dc:creator>Zhao, S. G.</dc:creator>
<dc:creator>Zwart, W.</dc:creator>
<dc:creator>Feng, F.</dc:creator>
<dc:creator>Quigley, D.</dc:creator>
<dc:creator>Lupien, M.</dc:creator>
<dc:date>2025-08-12</dc:date>
<dc:identifier>doi:10.1101/2025.08.08.668693</dc:identifier>
<dc:title><![CDATA[Genetic and Epigenetic Reprogramming of Transposable Elements Drives ecDNA-Mediated Metastatic Prostate Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.10.669490v1?rss=1">
<title>
<![CDATA[
The Hsp40 co-chaperone DNAJC7 modifies polyglutamine but not polyglycine aggregation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.10.669490v1?rss=1</link>
<description><![CDATA[
Protein-encoding nucleotide repeat expansion diseases, including polyglutamine (polyQ) and polyglycine (polyG) diseases, are characterized by the accumulation of aggregation-prone proteins. In the polyQ diseases, including Huntingtons disease and several spinocerebellar ataxias, substantial prior evidence supports a pathogenic role for mutant polyQ-expanded protein misfolding and aggregation, with molecular chaperones showing promise in suppressing disease phenotypes in cellular and animal models. The goal of this study is to establish a scalable cell-based model to systematically evaluate genetic modifiers of protein aggregation in both polyQ and polyG diseases. We developed FRET-based reporter systems that model polyQ and polyG aggregation in human cells and used them to perform high-throughput CRISPR interference screens targeting all known molecular chaperones. In the polyQ model, the screen identified multiple Hsp70 chaperones and Hsp40 co-chaperones previously implicated in polyQ aggregation and additionally revealed the Hsp40 co-chaperone DNAJC7 as a potent and previously unrecognized suppressor of polyQ aggregation. In contrast, in a FRET-based polyG aggregation model of neuronal intranuclear inclusion disease, CRISPRi screening showed minimal overlap of chaperone modifiers of the polyQ screen. Direct knockdown of DNAJC7 also did not affect polyG aggregation, yet overexpressed DNAJC7 co-localized with both polyQ and polyG aggregates in cells and reduced their aggregation. In addition to establishing new inducible, scalable cellular models for polyQ and polyG aggregation, this work expands the role of DNAJC7 in regulating folding of disease-associated proteins.
]]></description>
<dc:creator>Ramani, B.</dc:creator>
<dc:creator>Ehsani, K.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:date>2025-08-12</dc:date>
<dc:identifier>doi:10.1101/2025.08.10.669490</dc:identifier>
<dc:title><![CDATA[The Hsp40 co-chaperone DNAJC7 modifies polyglutamine but not polyglycine aggregation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.11.669701v1?rss=1">
<title>
<![CDATA[
Spatiotemporal control of cortical centrin patterning by regionalized Sfi1 family scaffolding proteins in Stentor coeruleus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.11.669701v1?rss=1</link>
<description><![CDATA[
Summary/AbstractPatterning is fundamental to the development and maintenance of organisms, ensuring functional and structural organization. While patterning is well studied at the level of multicellular organisms, even single cells need to undergo morphogenesis and form spatial patterns. Stentor coeruleus is a large ciliate that has been a classical system for studying patterning and morphogenesis due to its distinctive shape and organization of easily visible cortical structures which show a clear localization along the anterior-posterior body axis. The cortex of Stentor is made of two cytoskeletal layers, one composed of microtubules, and one composed of a network of centrin-family EF hand proteins, which form a branched network in the anterior half of the cell and long thick bundles known as myonemes in the posterior half. Sfi1 family proteins scaffold the assembly of centrin filaments throughout the eukaryotes, and function as scaffolding proteins in myonemal contractile systems found in some ciliates. A set of Sfi1 proteins upregulated during regeneration in Stentor were found to have a role in maintaining anterior/posterior differences in centrin patterning. Loss of this distinct anterior-posterior patterning leads to defects in regeneration and contraction. Using RNAi-mediated knockdown of Sfi1 genes, we have found that cells fail to regenerate their oral apparatus, with early expressed Sfi1 genes more critical for oral primordium (assembly of organized basal bodies that become the new oral apparatus) formation than those expressed later. Additionally, different Sfi1 proteins appear to be recruited sequentially to the growing primordium in an order that matches the time at which they are required for progression of regeneration. Knockdown of Sfi1 isoforms that results in reduced myoneme cables impairs contraction such that cells fail to contract in response to a stimulus or do not fully contract. These findings suggest a model in which regionalized differences in patterning of cytoskeletal assemblies can be modulated by regionalized localization of scaffolding proteins.
]]></description>
<dc:creator>Yan, C. J.</dc:creator>
<dc:creator>Steube, N.</dc:creator>
<dc:creator>Dey, G.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2025-08-13</dc:date>
<dc:identifier>doi:10.1101/2025.08.11.669701</dc:identifier>
<dc:title><![CDATA[Spatiotemporal control of cortical centrin patterning by regionalized Sfi1 family scaffolding proteins in Stentor coeruleus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.12.669895v1?rss=1">
<title>
<![CDATA[
Fine needle aspiration biopsy of breast specimens effectively harvests cells for patient-derived organoids modeling breast ductal carcinoma in situ 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.12.669895v1?rss=1</link>
<description><![CDATA[
BackgroundPatient-derived organoids (PDOs) generated from benign breast tissue and breast carcinomas have successfully recapitulated their respective in vivo counterparts. PDOs model tumorigenesis and allow for screening of novel therapeutics personalized to individual patients. However, acquiring cells to generate PDOs is cumbersome. We demonstrate the feasibility of fine needle aspiration biopsy (FNAB) for harvesting cells for PDOs modeling ductal carcinoma in situ (DCIS).

MethodsSurgical specimens from patients with biopsy-proven DCIS were used for this study. Core needle biopsy (CNB) was performed on fresh specimens in the operating room, and tissue was mechanically dissociated before culture in basement membrane extract (BME) and organoid medium to generate PDOs. FNAB was performed in the gross room on fresh specimens, and the remaining aspirate was similarly submitted for PDO culture.

ResultsPDOs were successfully generated in 15/18 specimens obtained by CNB and 7/11 specimens obtained by FNAB. The average time to initial organoid growth was 4 days for FNAB specimens compared to 19.3 days for CNB specimens. Tumor cells were seen on 7/11 FNAB smears and 16/18 CNB touch preps. Immunofluorescence staining confirmed the presence of both luminal and myoepithelial cells in derived PDOs.

ConclusionsFNAB effectively obtains cells for PDOs modeling DCIS. CNB after mincing yielded PDOs with a high success rate, but they were slow to establish. Notably, the time to organoid growth was significantly shorter for FNAB specimens. Thus, FNAB offers an efficient alternative for breast PDO culture and can reduce the time and resources spent on generating PDO cultures.
]]></description>
<dc:creator>Ye, J.</dc:creator>
<dc:creator>Goldhammer, N.</dc:creator>
<dc:creator>Vohra, P.</dc:creator>
<dc:creator>Warhadpande, S.</dc:creator>
<dc:creator>De Castro, G. C.</dc:creator>
<dc:creator>Maldonado Rodas, C.</dc:creator>
<dc:creator>Moasser, M. M.</dc:creator>
<dc:creator>Ramalingam, K.</dc:creator>
<dc:creator>Abe, S.</dc:creator>
<dc:creator>Alvarado, M.</dc:creator>
<dc:creator>Ewing, C.</dc:creator>
<dc:creator>Goodwin, K.</dc:creator>
<dc:creator>Mukhtar, R.</dc:creator>
<dc:creator>Wong, J.</dc:creator>
<dc:creator>Esserman, L.</dc:creator>
<dc:creator>Balassanian, R.</dc:creator>
<dc:creator>Rosenbluth, J. M.</dc:creator>
<dc:date>2025-08-13</dc:date>
<dc:identifier>doi:10.1101/2025.08.12.669895</dc:identifier>
<dc:title><![CDATA[Fine needle aspiration biopsy of breast specimens effectively harvests cells for patient-derived organoids modeling breast ductal carcinoma in situ]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.13.670183v1?rss=1">
<title>
<![CDATA[
Commensal taxa in gut microbiota limit antibiotic resistance during extended oral antibiotic use 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.13.670183v1?rss=1</link>
<description><![CDATA[
Certain bacterial infections, such as those involving prosthetics, can require antimicrobial therapy over months to years, potentially increasing the burden of antimicrobial resistance. Here we longitudinally track the antimicrobial resistome in mice during continuous antibiotic dosing over 21 months. The burden of antibiotic resistance genes (ARGs) initially increases, but, surprisingly, declines in later months, approaching levels observed in untreated animals. ARG burden is regulated by taxonomy and declines as ARG-harboring taxa that initially bloom are replaced by commensals. Furthermore, we find that the dynamics of antibiotic-induced ARG burden are influenced by age-related differences in microbial taxonomy and can be removed by fecal microbiota transplantation. We show that commensals may regulate the resistome by limiting the growth of ARG-harboring taxa, thereby providing antimicrobial expansion resistance.
]]></description>
<dc:creator>Cyphert, E. L.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Chu, V. T.</dc:creator>
<dc:creator>Dubey, A.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Zhong, Z.</dc:creator>
<dc:creator>Cockey, J. R.</dc:creator>
<dc:creator>Diaz, E. C. G.</dc:creator>
<dc:creator>Morales, A. L.</dc:creator>
<dc:creator>Nixon, J. C.</dc:creator>
<dc:creator>Garcia, M.</dc:creator>
<dc:creator>Zeng, S.</dc:creator>
<dc:creator>Rohatgi, S.</dc:creator>
<dc:creator>Wong, J.</dc:creator>
<dc:creator>Arjyal, R.</dc:creator>
<dc:creator>Mekonen, H.</dc:creator>
<dc:creator>Neff, N.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Shea, M. K.</dc:creator>
<dc:creator>Fu, X.</dc:creator>
<dc:creator>Booth, S. L.</dc:creator>
<dc:creator>Leifer, C. A.</dc:creator>
<dc:creator>Singh, A.</dc:creator>
<dc:creator>Langelier, C. R.</dc:creator>
<dc:creator>Hernandez, C. J.</dc:creator>
<dc:date>2025-08-13</dc:date>
<dc:identifier>doi:10.1101/2025.08.13.670183</dc:identifier>
<dc:title><![CDATA[Commensal taxa in gut microbiota limit antibiotic resistance during extended oral antibiotic use]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.11.669754v1?rss=1">
<title>
<![CDATA[
A population-scale atlas of blood and tissue in lupus nephritis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.11.669754v1?rss=1</link>
<description><![CDATA[
One Sentence SummaryA single-cell atlas of paired blood and tissue samples from Lupus Nephritis patients and healthy controls identified stromal and immune populations within renal tissue, including the scar-associated macrophage populations, which correlate with and may drive renal disease activity.

Lupus nephritis (LN), a severe manifestation of Systemic Lupus Erythematosus (SLE), is a heterogeneous disease driven by diverse immune and tissue cell types. We obtained 538K single-cell and 140K single-nuclear profiles from kidney biopsies of 155 LN patients and 30 pre-implantation transplant biopsy controls, along with 325K single-cell blood profiles overlapping many of these patients. We identified key tissue cell types and cell states, and immune cell states; we were able to determine cell states that were tissue specific, and those that were present in the blood. We observed that LN pathological features are significantly associated with cell states using differential gene expression and Covarying Neighborhood Analysis (CNA). These analyses revealed broad changes in cell states associated with irreversible chronic tissue damage. After controlling for the effects of ongoing tissue damage, we observed that expansion of key glomerular and Scar Associated Macrophages (SAMs) populations tracked with increasing inflammatory disease activity. SAMs appear to drive LN fibrosis and, in active disease, infiltrate the glomeruli more than other myeloid cells. These observations strongly support that therapeutic targeting of myeloid populations may offer an as-of-yet unproven strategy to prevent renal inflammation and ongoing kidney damage in LN.
]]></description>
<dc:creator>Gurajala, S.</dc:creator>
<dc:creator>Sugiarto, N. W.</dc:creator>
<dc:creator>Curtis, M.</dc:creator>
<dc:creator>Eisenhaure, T. M.</dc:creator>
<dc:creator>Arazi, A.</dc:creator>
<dc:creator>Fava, A.</dc:creator>
<dc:creator>Xiao, Q.</dc:creator>
<dc:creator>Mears, J.</dc:creator>
<dc:creator>Rovin, B.</dc:creator>
<dc:creator>Berthier, C. C.</dc:creator>
<dc:creator>Izmirly, P. M.</dc:creator>
<dc:creator>Barnas, J. L.</dc:creator>
<dc:creator>Hoover, P. J.</dc:creator>
<dc:creator>Peters, M.</dc:creator>
<dc:creator>Raychowdhury, R.</dc:creator>
<dc:creator>Horisberger, A.</dc:creator>
<dc:creator>Sakaue, S.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Furie, R. A.</dc:creator>
<dc:creator>Belmont, H. M.</dc:creator>
<dc:creator>Hildeman, D. A.</dc:creator>
<dc:creator>Woodle, E. S.</dc:creator>
<dc:creator>Dall'Era, M.</dc:creator>
<dc:creator>Putterman, C.</dc:creator>
<dc:creator>Kamen, D. L.</dc:creator>
<dc:creator>McMahon, M. A.</dc:creator>
<dc:creator>Grossman, J.</dc:creator>
<dc:creator>Kalunian, K. C.</dc:creator>
<dc:creator>Hodgin, J. B.</dc:creator>
<dc:creator>Payan-Schober, F.</dc:creator>
<dc:creator>Apruzzese, W.</dc:creator>
<dc:creator>Perlman, H.</dc:creator>
<dc:creator>Cuda, C. M.</dc:creator>
<dc:creator>Wofsy, D.</dc:creator>
<dc:creator>Guthridge, J. M.</dc:creator>
<dc:creator>Anolik, J. H.</dc:creator>
<dc:creator>James, J. A.</dc:creator>
<dc:creator>Accelerating Medicines Partnerships Rheumatoid Arthritis/Systemic Lu</dc:creator>
<dc:date>2025-08-14</dc:date>
<dc:identifier>doi:10.1101/2025.08.11.669754</dc:identifier>
<dc:title><![CDATA[A population-scale atlas of blood and tissue in lupus nephritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.14.670414v1?rss=1">
<title>
<![CDATA[
Cul5Wsb2 uses BCL2 proteins as co-receptors to target Bim for degradation. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.14.670414v1?rss=1</link>
<description><![CDATA[
Apoptosis helps eliminate damaged or unnecessary cells. Anti-apoptotic BCL2 family proteins protect cells by binding and inhibiting pro-apoptotic proteins. Our study reveals that BCL2 proteins also target Bim for degradation by serving to recruit Bim to the E3 ubiquitin ligase Cul5Wsb2. Wsb2 recognizes Bcl-xl through a motif conserved between Bcl-xl, Bcl-w and Bcl2, but not Mcl1. Disruption of this interaction through mutation of either Bcl-xl or Wsb2 blocks the binding of Wsb2 to the Bcl-xl/Bim dimer. Wsb2 also associates with the Mcl1/Bim dimer through a separate Wsb2 interface, suggesting that Wsb2 has evolved independent means to target Bim. While Wsb2 is not essential in most cells, it is essential in cells derived from tumors of the nervous system, and knockdown of Wsb2 in these lines causes death by apoptosis. This work uncovers a novel mechanism of apoptosis regulation, with implications for developing therapies against neuroblastomas and other cancers reliant on this pathway for survival.
]]></description>
<dc:creator>Vaysse-Zinkhofer, W.</dc:creator>
<dc:creator>Alcindor, E.</dc:creator>
<dc:creator>Garaffo, N.</dc:creator>
<dc:creator>Toczyski, D. P.</dc:creator>
<dc:date>2025-08-14</dc:date>
<dc:identifier>doi:10.1101/2025.08.14.670414</dc:identifier>
<dc:title><![CDATA[Cul5Wsb2 uses BCL2 proteins as co-receptors to target Bim for degradation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.14.667581v1?rss=1">
<title>
<![CDATA[
NIPBL dosage shapes genome folding by tuning the rate of cohesin loop extrusion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.14.667581v1?rss=1</link>
<description><![CDATA[
Genome folding is not static, but emerges from dynamic processes that control transcription, replication, recombination, and repair. DNA loop extrusion by cohesin is central to genome organization, yet it remains unclear how cells can tune extrusion kinetics to achieve precise and functional chromosome folding patterns. Here we discover extrusion rate acts as a tunable biophysical parameter in cells, quantitatively dialed by the respective dosage of the cohesin cofactors NIPBL and PDS5. Modulation of extrusion rate can offset changes in cohesin lifetime to buffer steady-state chromosome structure and transcriptional states, even in the face of abnormal extrusion dynamics. These findings provide a long-sought mechanistic basis for the genetic interactions between cohesin cofactors and the molecular origin of haploinsufficiency in cohesinopathies, such as Cornelia de Lange syndrome.
]]></description>
<dc:creator>Shah, R.</dc:creator>
<dc:creator>Tortora, M. M. C.</dc:creator>
<dc:creator>Louafi, N.</dc:creator>
<dc:creator>Rahmaninejad, H.</dc:creator>
<dc:creator>Hansen, K. L.</dc:creator>
<dc:creator>Anderson, E. C.</dc:creator>
<dc:creator>Wen, D.</dc:creator>
<dc:creator>Giorgetti, L.</dc:creator>
<dc:creator>Fudenberg, G.</dc:creator>
<dc:creator>Nora, E. P.</dc:creator>
<dc:date>2025-08-15</dc:date>
<dc:identifier>doi:10.1101/2025.08.14.667581</dc:identifier>
<dc:title><![CDATA[NIPBL dosage shapes genome folding by tuning the rate of cohesin loop extrusion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.14.669942v1?rss=1">
<title>
<![CDATA[
Multiplexed Perturbation Enables Scalable Pooled Screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.14.669942v1?rss=1</link>
<description><![CDATA[
CRISPR-based genetic perturbation screens have revolutionized the ability to link genes to cellular phenotypes with unprecedented precision and scale. However, conventional pooled CRISPR screens require large cell numbers to achieve adequate sgRNA representation, posing technical and financial challenges. Here, we investigate the impact of co-delivery of multiple guide RNAs via high multiplicity of infection (MOI) in pooled CRISPR interference (CRISPRi) screens as a strategy to enhance screening efficiency while reducing cell numbers. We systematically evaluate screen performance across varying MOIs, assessing the effects of multiplexing on knockdown efficiency, sgRNA representation, and potential interference of multiple sgRNA phenotypes. Our data demonstrate that sgRNA multiplexing (MOI 2.5-10) can maintain screen performance while enabling significant reductions in cell number requirements. We further apply these optimized conditions to conduct a genome-wide CRISPR screen for regulators of the intracellular adhesion molecule ICAM-1, successfully identifying novel candidates using as few as half a million cells. This study provides a framework for adopting multiplexed sgRNA strategies to streamline CRISPR screening applications in resource-limited settings.
]]></description>
<dc:creator>Oberlin, S.</dc:creator>
<dc:creator>Tay, N. Q.</dc:creator>
<dc:creator>Xue, A.</dc:creator>
<dc:creator>Pimentel, H.</dc:creator>
<dc:creator>McManus, M.</dc:creator>
<dc:date>2025-08-15</dc:date>
<dc:identifier>doi:10.1101/2025.08.14.669942</dc:identifier>
<dc:title><![CDATA[Multiplexed Perturbation Enables Scalable Pooled Screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.13.669976v1?rss=1">
<title>
<![CDATA[
Design of an ultrabright biosensor for dynamic imaging of kinase activity in cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.13.669976v1?rss=1</link>
<description><![CDATA[
Protein kinases regulate almost every major signaling pathway. Visualizing spatiotemporal dynamics of kinase activity is thus essential to understand cell signaling. Here we report a de novo-designed activity reporter of kinase, dubbed NOVARK, which contains a single polypeptide chain with multiple modular motifs that act as specific kinase substrates and reporters. NOVARK undergoes phosphorylation-induced higher order-assembly, which are detectable as ultrabright GFP droplets with a greater dynamic range than existing Forster resonance energy transfer-based kinase reporters. We designed versions of NOVARK that rapidly and reversibly report intracellular activity of protein kinase A, C, and ERK following stimulation/inhibition by upstream GPCR agonists. Our work provides a generalizable platform that enables the design of ultrabright biosensors for illuminating dynamic architecture of kinase signaling.
]]></description>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Tan, S. K.</dc:creator>
<dc:creator>Chung, C.-I.</dc:creator>
<dc:creator>Hatstat, K. A.</dc:creator>
<dc:creator>Zhao, Q.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:creator>shu, X.</dc:creator>
<dc:date>2025-08-16</dc:date>
<dc:identifier>doi:10.1101/2025.08.13.669976</dc:identifier>
<dc:title><![CDATA[Design of an ultrabright biosensor for dynamic imaging of kinase activity in cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.16.670679v1?rss=1">
<title>
<![CDATA[
An interneuronal CRH and CRHBP circuit stabilizes birdsong performance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.16.670679v1?rss=1</link>
<description><![CDATA[
The performance of skilled behaviors requires a balance between consistency and adaptability. Although the neural mechanisms that regulate this balance have been extensively studied at systems and physiological levels, relatively little is known about how the molecular properties of motor circuits influence motor stability versus flexibility. Here, we characterize the region- and cell-type specific expression patterns of neuropeptide systems across the neural circuit that controls the learning and performance of birdsong, a model for skilled behavior. We identify a number of neuropeptide pathways with differential expression between song regions and surrounding areas that are not involved in song production or learning. One of the strongest enriched genes in song regions is corticotropin releasing hormone binding protein (CRHBP), whose product binds corticotropin releasing hormone (CRH), a neuropeptide implicated in neuronal excitability and plasticity. We find that the expression of CRHBP in the song motor pathway decreases upon deafening-induced song destabilization, increases during song acquisition, and increases the more a bird sings. CRH and CRHBP are expressed in distinct interneuronal populations in song motor regions, providing a local neuromodulatory circuit well-positioned to regulate song performance. Consistent with this role, genetic and pharmacological manipulation of the CRH pathway in the song motor pathway resulted in bidirectional modifications of song variability, with elevated CRHBP acting to maintain low variability and elevated CRH acting to increase variability. These data indicate that an interneuronal neuropeptidergic pathway maintains the stability of song, acting as a local mechanism that regulates the balance between motor consistency versus flexibility.
]]></description>
<dc:creator>Colquitt, B. M.</dc:creator>
<dc:creator>Brainard, M.</dc:creator>
<dc:date>2025-08-17</dc:date>
<dc:identifier>doi:10.1101/2025.08.16.670679</dc:identifier>
<dc:title><![CDATA[An interneuronal CRH and CRHBP circuit stabilizes birdsong performance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.18.670983v1?rss=1">
<title>
<![CDATA[
Bivalent mRNA booster encoding virus-like particles elicits potent polyclass RBD antibodies in pre-vaccinated mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.18.670983v1?rss=1</link>
<description><![CDATA[
mRNA vaccines emerged as a leading vaccine technology during the COVID-19 pandemic. However, their sustained protective efficacies were limited by relatively short-lived antibody responses and the emergence of SARS-CoV-2 variants, necessitating frequent and variant-updated boosters. We recently developed the ESCRT- and ALIX-binding region (EABR) mRNA vaccine platform, which encodes engineered immunogens that induce budding of enveloped virus-like particles (eVLPs) from the plasma membrane, thereby resulting in presentation of immunogens on cell surfaces and eVLPs. Prior studies showed that spike (S)-EABR mRNA-LNP immunizations elicited enhanced neutralizing antibody responses against ancestral and variant SARS-CoV-2 compared with conventional S mRNA-LNP in naive mice, but the effectiveness of S-EABR mRNA-LNP boosters in the context of pre-existing immunity has not been investigated. Here, we evaluated monovalent Wuhan-Hu-1 (Wu1) and bivalent (Wu1/BA.5) S-EABR mRNA-LNP boosters in mice pre-vaccinated with conventional Wu1 S mRNA-LNP. Compared to conventional S mRNA-LNP boosters, the EABR approach enhanced monovalent and bivalent mRNA-LNP booster-induced neutralizing responses against Omicron subvariants BA.1, BA.5, BQ.1.1, and XBB.1, with bivalent S-EABR mRNA-LNP consistently eliciting the highest titers. Epitope mapping of polyclonal antisera by deep mutational scanning revealed that bivalent S-EABR mRNA-LNP boosted diverse "polyclass" anti-receptor-binding domain (RBD) responses, suggesting balanced targeting of multiple RBD epitope classes. In contrast, monovalent S, bivalent S, and monovalent S-EABR mRNA-LNP boosters elicited less diverse polyclonal serum responses primarily targeting immunodominant RBD epitopes. Cryo-EM structures demonstrated that bivalent mRNA immunizations promote S heterotrimer formation, potentially enhancing bivalent S-EABR mRNA-LNP booster-induced antibody breadth and polyclass epitope targeting by activating cross-reactive B cells through intra-S crosslinking. These findings support the future design of bivalent or multivalent S-EABR mRNA-LNP boosters as a promising strategy to confer broader, and therefore potentially more durable, protection against emerging SARS-CoV-2 variants and other rapidly evolving viruses.
]]></description>
<dc:creator>Fan, C.</dc:creator>
<dc:creator>Cohen, A. A.</dc:creator>
<dc:creator>Dam, K.-M. A.</dc:creator>
<dc:creator>Rorick, A. V.</dc:creator>
<dc:creator>Priso Fils, A.-C. I.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Gnanapragasam, P. N.</dc:creator>
<dc:creator>Segovia, L. N.</dc:creator>
<dc:creator>Huey-Tubman, K. E.</dc:creator>
<dc:creator>Moon, W. J.</dc:creator>
<dc:creator>Lin, P. J.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Hoffmann, M. A.</dc:creator>
<dc:date>2025-08-19</dc:date>
<dc:identifier>doi:10.1101/2025.08.18.670983</dc:identifier>
<dc:title><![CDATA[Bivalent mRNA booster encoding virus-like particles elicits potent polyclass RBD antibodies in pre-vaccinated mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.18.670970v1?rss=1">
<title>
<![CDATA[
White Matter Tract Vulnerability to Amyloid Pathology on the Alzheimer's Disease Continuum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.18.670970v1?rss=1</link>
<description><![CDATA[
Alzheimers disease (AD) is marked by progressive cognitive decline and memory loss, due to the abnormal accumulation of amyloid-beta (A{beta}) plaques, which facilitate the spread of tau pathology, and a gradually spreading pattern of neuronal loss. Understanding how amyloid positivity affects the brains neural pathways is critical for understanding how the brain changes with AD pathology. Tractometry offers a powerful approach for the in vivo, 3D quantitative assessment of white matter tracts, enabling the localization of microstructural abnormalities in diseased populations and those at risk. In this study, we applied BUAN (Bundle Analytics) tractometry to multi-cohort diffusion MRI data from a total of 1,908 participants: 606 participants in ADNI3 (Alzheimers Disease Neuroimaging Initiative Phase 3) and 1,302 participants from the HABS-HD (Health and Aging Brain Study-Health Disparities). Using BUAN and along-tract statistical analysis, we assessed the localized effects of amyloid positivity on white-matter pathways, which may be further influenced by downstream tau accumulation. Amyloid positivity was quantified via amyloid-sensitive positron emission tomography (PET). BUAN enables tract-specific quantification of white matter microstructure and supports statistical testing along the full length of fiber bundles to detect subtle, spatially localized associations. We present 3D visualizations of tractwise amyloid associations, highlighting distinct patterns of white matter degeneration in AD.
]]></description>
<dc:creator>Chandio, B. Q.</dc:creator>
<dc:creator>Nir, T. M.</dc:creator>
<dc:creator>Villalon-Reina, J. E.</dc:creator>
<dc:creator>Thomopoulos, S. I.</dc:creator>
<dc:creator>Feng, Y.</dc:creator>
<dc:creator>Reid, R. I.</dc:creator>
<dc:creator>Jack, C. R.</dc:creator>
<dc:creator>Weiner, M. W.</dc:creator>
<dc:creator>Garyfallidis, E.</dc:creator>
<dc:creator>Jahanshad, N.</dc:creator>
<dc:creator>Braskie, M. N.</dc:creator>
<dc:creator>O'Bryant, S.</dc:creator>
<dc:creator>Thompson, P. M.</dc:creator>
<dc:date>2025-08-20</dc:date>
<dc:identifier>doi:10.1101/2025.08.18.670970</dc:identifier>
<dc:title><![CDATA[White Matter Tract Vulnerability to Amyloid Pathology on the Alzheimer's Disease Continuum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.19.671124v1?rss=1">
<title>
<![CDATA[
Regulated development of cannibalistic supergiant cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.19.671124v1?rss=1</link>
<description><![CDATA[
Virtually all paradigms in developmental biology apply to differentiating cells and tissues within multicellular animals and plants. However, unicellular eukaryotes, which must simultaneously perform all roles necessary for organismal function, also form complex and specialized structures, using processes that take place exclusively within the confines of a single cell. Here, we describe a ciliate (Euplotes gigatrox sp. nov.) undergoing drastic morphological transformations within a genetically uniform population, the most spectacular being the appearance of "supergiants" that increase in size, change shape, and modify their locomotion and feeding behaviour to cannibalize clonal relatives. We explore supergiant formation from the perspective of life cycle, ecological strategy, and gene expression, demonstrating that supergiants are distinct, regulated, transcriptionally unique stages. These reversibly differentiated cells require both external and internal triggers to develop and have evolved regulatory loops to ensure coupling between environmental and physiological conditions. This system provides a blueprint for approaching both cell differentiation and functional ecology in unicellular organisms, which might open new avenues for the generalization and contextualization of known morphogenetic mechanisms, as well as the discovery of new ones.
]]></description>
<dc:creator>Larson, B. T.</dc:creator>
<dc:creator>Giannotti, D.</dc:creator>
<dc:creator>Mtawali, M.</dc:creator>
<dc:creator>Lord, S. J.</dc:creator>
<dc:creator>Boscaro, V.</dc:creator>
<dc:creator>Keeling, P. J.</dc:creator>
<dc:date>2025-08-20</dc:date>
<dc:identifier>doi:10.1101/2025.08.19.671124</dc:identifier>
<dc:title><![CDATA[Regulated development of cannibalistic supergiant cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.21.671373v1?rss=1">
<title>
<![CDATA[
Divergent viral phosphodiesterases for immune signaling evasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.21.671373v1?rss=1</link>
<description><![CDATA[
Cyclic dinucleotides (CDNs) and other short oligonucleotides play fundamental roles in immune system activation in organisms ranging from bacteria to humans. In response, viruses use phosphodiesterase-mediated oligonucleotide cleavage for immune evasion, a strategy whose diversity has not yet been explored. We used a canonical 2H phosphodiesterase (2H PDE) structure-based search of prokaryotic and eukaryotic viral sequences to identify an exceptional diversity of 2H PDEs across the virome, including enzymes not detectable with sequence search methods alone. Despite active site conservation, biochemical experiments revealed remarkable substrate specificity of these PDEs that corresponds to variation in the core 2H fold. This nuanced specificity allows 2H PDEs to selectively degrade oligonucleotide messengers to avoid interfering with host immune signaling. Together, these findings nominate viral 2H PDEs as key regulators of CDN signaling across the tree of life.
]]></description>
<dc:creator>Doherty, E. E.</dc:creator>
<dc:creator>Nomburg, J. E.</dc:creator>
<dc:creator>Adler, B. A.</dc:creator>
<dc:creator>Lopez, S.</dc:creator>
<dc:creator>Hsieh, K.</dc:creator>
<dc:creator>Price, N.</dc:creator>
<dc:creator>Blount, N.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:date>2025-08-21</dc:date>
<dc:identifier>doi:10.1101/2025.08.21.671373</dc:identifier>
<dc:title><![CDATA[Divergent viral phosphodiesterases for immune signaling evasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.18.670802v1?rss=1">
<title>
<![CDATA[
Inhibitory NK receptors regulate the γδ T cell response to malaria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.18.670802v1?rss=1</link>
<description><![CDATA[
Gamma delta ({gamma}{delta}) T cells are important mediators of the immune response to childhood malaria infection. Human V{gamma}9+V{delta}2+ T cells possess intrinsic, HLA-independent responsiveness to Plasmodium falciparum phosphoantigens produced in the blood stage of malaria infection. Engagement of the {gamma}{delta} T cell receptor (TCR) by phosphoantigen-bound butyrophilin molecules results in V{gamma}9+V{delta}2+ T cell expansion, pro-inflammatory cytokine production, and release of cytotoxic granules that mediate parasite killing. Repeated malaria infection, however, leads to a reduction in circulating V{gamma}9+V{delta}2+ T cells and upregulation of immunomodulatory molecules, including NK receptors, that correlates with less severe symptoms upon infection. We explore phenotypic and functional differences of {gamma}{delta} T cells in Ugandan children with high versus low malaria exposure, utilizing high-parameter spectral flow cytometry analysis of PBMCs. We observed significant differences in expression of inhibitory NK receptors - KIR2DL1, KIR2DL2/3, KIR3DL1, LILRB1, and NKG2A - on {gamma}{delta} T cell subsets, with V{gamma}9+V{delta}2+ T cells exhibiting a divergent mechanism of control compared to other subsets. We found that NKG2A and KIR3DL1 expression associated with potent V{gamma}9+V{delta}2+ T cell responses to TCR- and Fc receptor (FcR)-mediated stimulation while KIR2DL1, KIR2DL2/3 and LILRB1 associated with reduced degranulation and cytokine production. These results identify a new role for inhibitory NK receptors expressed on {gamma}{delta} T cells, exerting a finely tuned balance of activating and inhibitory signals to regulate the response to malaria-related antigens.

AUTHOR SUMMARYMalaria remains one of the deadliest infectious diseases, disproportionately affecting young children in sub-Saharan Africa who succumb to sequelae of Plasmodium falciparum infection. However, children living in highly endemic areas experience repeated malaria infection and develop naturally acquired-but not sterilizing-immunity which leads to an asymptomatic reinfection pattern. The immune factors that determine the balance of inflammatory and tolerogenic functions seen in non-sterilizing malaria immunity are yet to be fully understood. Here, we focus on the phenotypic and functional differences in one cell type between children with a history of low versus high malaria exposure. We identified a group of inhibitory surface receptors that improved the antimalarial function of this cell, and another group that worsened their function. Our study clarifies the immune landscape in highly malaria-exposed individuals and illuminates one potential system of regulating the cellular response to repeat infection.
]]></description>
<dc:creator>Olive, M. E.</dc:creator>
<dc:creator>Callaway, P. C.</dc:creator>
<dc:creator>Ilala, M.</dc:creator>
<dc:creator>Levan, J.</dc:creator>
<dc:creator>Acevedo, G. R.</dc:creator>
<dc:creator>Nankya, F.</dc:creator>
<dc:creator>Arinaitwe, E.</dc:creator>
<dc:creator>Rek, J.</dc:creator>
<dc:creator>Jagannathan, P.</dc:creator>
<dc:creator>Dorsey, G.</dc:creator>
<dc:creator>Kamya, M. R.</dc:creator>
<dc:creator>Feeney, M. E.</dc:creator>
<dc:date>2025-08-22</dc:date>
<dc:identifier>doi:10.1101/2025.08.18.670802</dc:identifier>
<dc:title><![CDATA[Inhibitory NK receptors regulate the γδ T cell response to malaria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.19.670119v1?rss=1">
<title>
<![CDATA[
Structure-guided engineering of CCL27 enhances natural ligand CAR T-cells against CCR10 for multiple myeloma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.19.670119v1?rss=1</link>
<description><![CDATA[
Despite the success of BCMA CAR-Ts, many multiple myeloma patients relapse and require additional therapeutic options. Our group previously identified the chemokine receptor CCR10 as a potential alternate target to address this need. Here, we validated CCR10 expression on primary myeloma tumors and sought to develop CAR T-cells against CCR10, utilizing its natural ligand CCL27 as a CAR binding element. However, CARs based on the native CCL27 sequence were ineffective. We thus utilized computational modeling and structure-guided engineering to inform rational mutations along the CCL27-CCR10 interface, exploiting a hydrophobic pocket on CCR10. This effort identified CCL27 mutants with an additional N-terminal aromatic amino acid that dramatically improved the efficacy of CCL27-based CAR-Ts to near that of current anti-BCMA CAR-Ts. We validated key amino acid contacts at the CCL27-CCR10 interface, which contribute to increased CAR binding avidity, predicted to be influenced by increased Van der Waals interactions. Lastly, we found that the CCL27 mutants have no toxicity in the hematopoietic compartment. This work illustrates the potential of engineering natural ligand CAR-Ts beyond their wild-type sequences and underscores the translational potential of engineered CCL27 mutant CAR-Ts.
]]></description>
<dc:creator>Chilakapati, N.</dc:creator>
<dc:creator>Patino-Escobar, B.</dc:creator>
<dc:creator>Chen, E. Y.</dc:creator>
<dc:creator>Dalal, R.</dc:creator>
<dc:creator>de Montagnac, J.</dc:creator>
<dc:creator>Johnson, H.</dc:creator>
<dc:creator>Kang, A. S.</dc:creator>
<dc:creator>Akagi, N.</dc:creator>
<dc:creator>Ramos, E.</dc:creator>
<dc:creator>Phojanakong, P.</dc:creator>
<dc:creator>Salangsang, F.</dc:creator>
<dc:creator>Zeng, Y.</dc:creator>
<dc:creator>Chari, A.</dc:creator>
<dc:creator>Chung, A.</dc:creator>
<dc:creator>Kumar, A. D.</dc:creator>
<dc:creator>Martin, T. G.</dc:creator>
<dc:creator>Shy, B. R.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Karlon, W. J.</dc:creator>
<dc:creator>Kortemme, T.</dc:creator>
<dc:creator>Barwick, B. G.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:date>2025-08-23</dc:date>
<dc:identifier>doi:10.1101/2025.08.19.670119</dc:identifier>
<dc:title><![CDATA[Structure-guided engineering of CCL27 enhances natural ligand CAR T-cells against CCR10 for multiple myeloma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.19.670946v1?rss=1">
<title>
<![CDATA[
PHLDA2 promotes breast cancer metastasis by co-opting a developmental program for placental vascular remodeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.19.670946v1?rss=1</link>
<description><![CDATA[
Identifying drivers of metastasis is essential for developing new treatments for patients with advanced disease. Here, we identify PHLDA2 as a robust driver of breast cancer metastasis. Previous work established PHLDA2 as an imprinted gene expressed by trophoblasts which are critical for vascular remodeling during placental development. We find that hypomethylation of PHLDA2 in breast tumors correlates with increased gene expression, which is associated with metastasis and poor survival in breast cancer patients. RNA-sequencing showed that PHLDA2 overexpression results in upregulation of genes that control invasion, extracellular matrix assembly, and vascular remodeling, consistent with trophoblast functions in placental development. Using an in vitro vascularized microtumor (VMT) system, we find that PHLDA2 functions through SPARC, which promotes metastasis by inducing vascular permeability and enhancing tumor dissemination. These data suggest that increased expression of PHLDA2 through hypomethylation promotes metastasis by ectopic expression of a developmental program for vascular remodeling.
]]></description>
<dc:creator>Halas, P. V.</dc:creator>
<dc:creator>Savage, H.</dc:creator>
<dc:creator>Hachey, S. J.</dc:creator>
<dc:creator>Lev, T.</dc:creator>
<dc:creator>Prem Antony Samy, A. L.</dc:creator>
<dc:creator>Chan, C.</dc:creator>
<dc:creator>Jimenez, S.</dc:creator>
<dc:creator>Domadia, R. M.</dc:creator>
<dc:creator>Lin, I.</dc:creator>
<dc:creator>Mallya, S.</dc:creator>
<dc:creator>Insua-Rodriguez, J.</dc:creator>
<dc:creator>Gonzalez, J.</dc:creator>
<dc:creator>Adam, I.</dc:creator>
<dc:creator>Alshetaiwi, H.</dc:creator>
<dc:creator>Hernandez, G. A.</dc:creator>
<dc:creator>Longworth, A.</dc:creator>
<dc:creator>McMullen, T. P.</dc:creator>
<dc:creator>Kessenbrock, K.</dc:creator>
<dc:creator>Hughes, C. C.</dc:creator>
<dc:creator>Ma, D.</dc:creator>
<dc:creator>Lawson, D. A.</dc:creator>
<dc:date>2025-08-23</dc:date>
<dc:identifier>doi:10.1101/2025.08.19.670946</dc:identifier>
<dc:title><![CDATA[PHLDA2 promotes breast cancer metastasis by co-opting a developmental program for placental vascular remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.21.671605v1?rss=1">
<title>
<![CDATA[
HP1B and H3K9me3 Regulate Olfactory Receptor Choice and Transcriptional Identity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.21.671605v1?rss=1</link>
<description><![CDATA[
Diverse epigenetic regulatory mechanisms ensure and regulate cellular diversity. Among others, the histone 3 lysine 9 me3 (H3K9me3) post translational modification participates in silencing lineage-inappropriate genes. H3K9me3 restricts access of transcription factors and other regulatory proteins to cell-fate controlled genes. In mice, olfactory sensory neurons (OSN) express one olfactory receptor (OR) gene out of 2,600 possibilities. This monoallelic and stochastic OR choice happens as OSNs differentiate and undergo dramatic changes in nuclear architecture. OR genes from different chromosomes converge into specialized nuclear bodies and chromatin compartments as H3K9me3 and chromatin binding proteins including heterochromatin protein 1 (HP1) are incorporated. In this work, we have uncovered an unexpected role for HP1{beta} in OR choice and neuronal identity that cannot be rescued by HP1 in vivo. With the use of a conditional knock-in mouse model that replaces HP1{beta} for HP1, we observe changes in H3K9me3 levels, DNA accessibility, and Hi-C contacts over OR gene clusters. These changes alter the expression patterns that partition the mouse olfactory epithelium into five OR expression zones, which results in a reduced OR repertoire leading to a loss of olfactory sensory neuron diversity. We propose that HP1{beta} modulates the competition of OR-promoters for enhancers to promote receptor diversity, by establishing repression gradients in a zonal fashion.
]]></description>
<dc:creator>Escamilla-del-Arenal, M.</dc:creator>
<dc:creator>Duffie, R.</dc:creator>
<dc:creator>Shayya, H.</dc:creator>
<dc:creator>Loconte, V.</dc:creator>
<dc:creator>Ekman, A.</dc:creator>
<dc:creator>Street, L. A.</dc:creator>
<dc:creator>Horta, A.</dc:creator>
<dc:creator>Canzio, D.</dc:creator>
<dc:creator>Monahan, K.</dc:creator>
<dc:creator>Larabell, C. A.</dc:creator>
<dc:creator>Jovanovic, M.</dc:creator>
<dc:creator>Lomvardas, S.</dc:creator>
<dc:date>2025-08-25</dc:date>
<dc:identifier>doi:10.1101/2025.08.21.671605</dc:identifier>
<dc:title><![CDATA[HP1B and H3K9me3 Regulate Olfactory Receptor Choice and Transcriptional Identity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.25.668405v1?rss=1">
<title>
<![CDATA[
A fibroblast-centric network drives cold fibrosis in the tumor microenvironment of lung squamous cell carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.25.668405v1?rss=1</link>
<description><![CDATA[
The tumor microenvironment (TME) of chronic inflammation-associated cancers (CIACs) is shaped by cycles of injury and maladaptive repair, yet the principles organizing fibrotic stroma in these tumors remain unclear. Here, we applied the concept of hot versus cold fibrosis, originally credentialed in non-cancerous fibrosis of heart and kidney, to lung squamous cell carcinoma (LUSC), a prototypical CIAC. Single-cell transcriptomics of matched tumor and adjacent-normal tissue from 16 treatment-naive LUSC patients identified a cold fibrotic architecture in the LUSC TME: cancer-associated fibroblasts (CAFs) expanded and adopted myofibroblast and stress-response states, while macrophages were depleted. This macrophage-poor, CAF-rich stroma was maintained by CAF autocrine growth factor loops, including TIMP1, INHBA, TGFB1, and GMFB. In parallel, the immune compartment exhibited a hot tumor phenotype with abundant T and B cells, forming spatially distinct but molecularly engaged networks with CAFs. CAF gene programs typifying cold fibrosis in LUSC were conserved in other CIACs, including esophageal and gastric adenocarcinomas. These results redefine desmoplastic regions of tumors through the lens of a non-cancer fibrosis model, demonstrating that conserved stromal circuits constitute therapeutic vulnerabilities in CIACs.
]]></description>
<dc:creator>Miyara, S.</dc:creator>
<dc:creator>Frenkel, S.</dc:creator>
<dc:creator>Mayo, A.</dc:creator>
<dc:creator>Gascard, P.</dc:creator>
<dc:creator>Strasser, M.</dc:creator>
<dc:creator>Gibbs, D.</dc:creator>
<dc:creator>Weizman, E.</dc:creator>
<dc:creator>Ben Shalom, I.</dc:creator>
<dc:creator>Stein, Y.</dc:creator>
<dc:creator>Pan, D.</dc:creator>
<dc:creator>Caruso, J. A.</dc:creator>
<dc:creator>Sangwan, V.</dc:creator>
<dc:creator>Bertos, N.</dc:creator>
<dc:creator>Berube, J.</dc:creator>
<dc:creator>Camilleri-Broet, S.</dc:creator>
<dc:creator>Oikonomopoulos, S.</dc:creator>
<dc:creator>Djambazian, H.</dc:creator>
<dc:creator>Umansky, K. B.</dc:creator>
<dc:creator>Elkahal, J.</dc:creator>
<dc:creator>Mayer, S.</dc:creator>
<dc:creator>Fiset, P. O.</dc:creator>
<dc:creator>Adler, M.</dc:creator>
<dc:creator>Tzahor, E.</dc:creator>
<dc:creator>Ragoussis, I.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Ferri, L.</dc:creator>
<dc:creator>Tlsty, T.</dc:creator>
<dc:creator>Scherz-Shouval, R.</dc:creator>
<dc:creator>Alon, U.</dc:creator>
<dc:date>2025-08-25</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.668405</dc:identifier>
<dc:title><![CDATA[A fibroblast-centric network drives cold fibrosis in the tumor microenvironment of lung squamous cell carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.21.671671v1?rss=1">
<title>
<![CDATA[
Integrated ambient modeling and genetic demultiplexing of single-cell RNA+ATAC multiome experiments with Ambimux 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.21.671671v1?rss=1</link>
<description><![CDATA[
Single cell technologies have advanced at a rapid pace, providing assays for various molecular phenotypes. Droplet-based single cell technologies, particularly those based on nuclei isolation, such as simultaneous RNA+ATAC single-cell multiome, are susceptible to exogenous ambient molecule contamination, which can increase noise in cell type-level associations. We reasoned that genotype-based sample multiplexing can provide an opportunity to infer this ambient contamination by leveraging DNA variation in sequenced reads. Thus, we developed ambimux, a likelihood-based method to estimate ambient fractions and demultiplex single-cell multiome experiments using genotype-level data. Ambimux models the ambient or nuclear probability at the read level and thus can classify empty droplets and estimate droplet-specific ambient molecule fractions in each modality. We first evaluated our method using simulated data sets across a range of parameters. We found that ambimux closely estimated the ground truth droplet contamination fractions in the RNA (MAE=0.048) and ATAC (MAE=0.042) modalities. As a result, ambimux maintained high specificity (>95%) and was able to correctly assign singlets at considerably high ambient fractions (up to 60%) for both RNA and ATAC modalities. In comparison with models that do not consider ambient contamination, these only maintained similar sensitivity levels at considerably lower ambient fractions (up to 25%). We then generated a real data set of seven visceral adipose tissue biopsies run on a single 10x Multiome channel. We ran ambimux and detected 4,986 singlets, capturing similar numbers as other methods.

Then, we sought to evaluate the fidelity of the ambient fraction estimates from ambimux. We split singlets into ambient-enriched (>5% contamination in both modalities) or nuclear-enriched (<5% in both) droplets and performed gene-peak linkage analysis. Low ambient droplets resulted in more significant hits with gene-peak links enriched at the transcription start site relative to high ambient droplets, suggesting that the ambient droplets identified by ambimux hamper the identification of biologically meaningful signals. In summary, we developed a joint single-cell multiome demultiplexing method, ambimux, that accurately models and estimates ambient molecule contamination in each modality.
]]></description>
<dc:creator>Alvarez, M.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Lee, S. H. T.</dc:creator>
<dc:creator>Arasu, U. T.</dc:creator>
<dc:creator>Selvarajan, I.</dc:creator>
<dc:creator>Örd, T.</dc:creator>
<dc:creator>Rahmani, E.</dc:creator>
<dc:creator>Chen, Z. J.</dc:creator>
<dc:creator>Avram, O.</dc:creator>
<dc:creator>Kar, A.</dc:creator>
<dc:creator>Kaminska, D.</dc:creator>
<dc:creator>Männistö, V.</dc:creator>
<dc:creator>Halperin, E.</dc:creator>
<dc:creator>Pihlajamäki, J.</dc:creator>
<dc:creator>Luo, C.</dc:creator>
<dc:creator>Kaikkonen, M. U.</dc:creator>
<dc:creator>Zaitlen, N.</dc:creator>
<dc:creator>Pajukanta, P.</dc:creator>
<dc:date>2025-08-26</dc:date>
<dc:identifier>doi:10.1101/2025.08.21.671671</dc:identifier>
<dc:title><![CDATA[Integrated ambient modeling and genetic demultiplexing of single-cell RNA+ATAC multiome experiments with Ambimux]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.24.671323v1?rss=1">
<title>
<![CDATA[
Substitution spectrum and selection at G-quadruplexes in great ape telomere-to-telomere genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.24.671323v1?rss=1</link>
<description><![CDATA[
G-quadruplexes (G4s) are noncanonical DNA secondary structures formed by runs of guanines (stems) connected by other nucleotides (loops). These structures are enriched at regulatory regions such as promoters, CpG islands, untranslated regions (UTRs), enhancers, and replication origins, where they play key roles in transcription and replication. Although prior studies have demonstrated that G4s exhibit higher mutation rates than canonical DNA, little is known about the substitution patterns and selection acting specifically on G4 stems and loops. In this study, we utilized Telomere-to-Telomere (T2T) genome assemblies from human and two non-human great apes (chimpanzee and Bornean orangutan) to analyze substitution spectra and selective constraints within G4s, focusing on differences between stems and loops. We observed that fixed nucleotide substitutions leading to the gain or loss of G4 structures are more frequently located at stems, while those in G4s conserved across species are more often found at loops. On the other hand, single nucleotide polymorphisms had higher frequencies at stems than loops for all G4s, with a particularly high difference for singleton polymorphisms, suggesting higher mutation rates at stems than loops. To evaluate selection, we employed two approaches: we computed the ratio of substitution to polymorphism frequencies at stems vs. loops and performed phylogenetic modeling using PhyloFit. Both methods consistently revealed that stems of shared G4s experience stronger purifying selection than loops, particularly at promoters, enhancers, and UTRs. Our results provide novel insights into the sequence variation and selection of G4s, informing our understanding of their contributions to genome evolution and function.

Significance StatementG-quadruplexes (G4s) are non-canonical DNA structures that influence transcription, genome stability, and epigenetic regulation, yet their evolutionary dynamics in primates remain poorly understood. Leveraging recent T2T genome assemblies, we conducted a sequence-level analysis of G4 evolution across three ape lineages and used two methods to infer selection in G4. Shared G4s and species-specific G4s display distinct evolutionary signatures. Using PhyloFit to estimate substitution rates and the Hudson-Kreitman-Aguade test to contrast divergence with polymorphism, we found stems under markedly stronger purifying selection than loops, especially within promoters, CpG islands, and 5'UTRs. This pattern indicates that maintaining stem integrity is functionally critical and evolutionarily conserved. Our findings reveal how selective constraints vary both within G4 motifs and across genomic landscapes, offering insights for future studies on their functional importance and structural stability.
]]></description>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Mohanty, S. K.</dc:creator>
<dc:creator>Chiaromonte, F.</dc:creator>
<dc:creator>Makova, K.</dc:creator>
<dc:date>2025-08-27</dc:date>
<dc:identifier>doi:10.1101/2025.08.24.671323</dc:identifier>
<dc:title><![CDATA[Substitution spectrum and selection at G-quadruplexes in great ape telomere-to-telomere genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.27.672545v1?rss=1">
<title>
<![CDATA[
Using DNA origami to study nanoscale organization of plasma membranes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.27.672545v1?rss=1</link>
<description><![CDATA[
Plasma membrane (PM) lipids and proteins are organized into nanoscale regions called nanodomains, which regulate essential cellular processes by controlling local membrane organization. Despite advances in super-resolution microscopy and single particle tracking, the small size and temporal instability of nanodomains make them difficult to study in living cells. To overcome these challenges, we built fluorescent DNA origami probes that insert into the PM via lipid anchors displayed on the cell. The number and spatial distribution of anchors between the origami and the cell surface were precisely defined by the origami, enabling nanometer-scale sampling of the cell surface. Inserting these DNA origami particles into the membrane with lipid anchors allowed them to passively diffuse across the membrane, and we tracked their movement using single particle tracking to survey the PM landscape. By varying the number and spatial arrangement of lipid anchors connecting the DNA origami to the cell surface, we showed that immobilization of DNA origami particles requires simultaneous interactions with multiple nanodomains. Disruption of the actin cytoskeleton reduced immobilization, confirming its role in supporting nanodomain stability. Moreover, transient mechanical stretching of cells led to reversible increases in DNA origami mobility, indicating that mechanical force can reversibly regulate PM nanodomain organization. Altogether, we present a novel membrane-integrated DNA origami approach that provides mechanistic insights into PM nanodomain architecture and dynamics in living cells.
]]></description>
<dc:creator>Corradi, E.</dc:creator>
<dc:creator>Shen, K.</dc:creator>
<dc:creator>Karatas, Z.</dc:creator>
<dc:creator>Cercy, M.</dc:creator>
<dc:creator>Schlichthaerle, T.</dc:creator>
<dc:creator>Osouf, M.</dc:creator>
<dc:creator>Vialet, B.</dc:creator>
<dc:creator>Barthelemy, P.</dc:creator>
<dc:creator>Rosendale, M.</dc:creator>
<dc:creator>Radhakrishnan, A. V.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Jungmann, R.</dc:creator>
<dc:creator>Gissot, A.</dc:creator>
<dc:creator>Douglas, S.</dc:creator>
<dc:creator>Giannone, G.</dc:creator>
<dc:date>2025-08-27</dc:date>
<dc:identifier>doi:10.1101/2025.08.27.672545</dc:identifier>
<dc:title><![CDATA[Using DNA origami to study nanoscale organization of plasma membranes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.22.671829v1?rss=1">
<title>
<![CDATA[
DTI versus NODDI White Matter Microstructural Biomarkers of Alzheimer's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.22.671829v1?rss=1</link>
<description><![CDATA[
Diffusion MRI (dMRI) is a powerful tool to assess white matter (WM) microstructural abnormalities in Alzheimers disease (AD). The fourth phase of the Alzheimers Disease Neuroimaging Initiative (ADNI) now includes multiple multishell dMRI protocols, enabling both traditional and advanced dMRI model analyses. There is a need to evaluate whether multishell data offer deeper insights into WM pathology in AD than more widely available single-shell data by overcoming single-shell model limitations. Here, we fit single-shell DTI and multishell NODDI to dMRI data from 533 ADNI3/4 participants to assess their sensitivity to key clinical indicators of AD such as cognitive impairment, amyloid-beta and tau PET burden. Overall, we found that NODDI offered no major advantages in detecting cognitive impairment and tau pathology, but NODDI was marginally more sensitive to amyloid pathology.
]]></description>
<dc:creator>Liou, K.</dc:creator>
<dc:creator>Thomopoulos, S. I.</dc:creator>
<dc:creator>Yoo, H.</dc:creator>
<dc:creator>Shuai, Y.</dc:creator>
<dc:creator>Chehrzadeh, S.</dc:creator>
<dc:creator>Arani, A.</dc:creator>
<dc:creator>Borowski, B.</dc:creator>
<dc:creator>Reid, R. I.</dc:creator>
<dc:creator>Jack, C. R.</dc:creator>
<dc:creator>Weiner, M. W.</dc:creator>
<dc:creator>Jahanshad, N.</dc:creator>
<dc:creator>Thompson, P. M.</dc:creator>
<dc:creator>Nir, T. M.</dc:creator>
<dc:date>2025-08-28</dc:date>
<dc:identifier>doi:10.1101/2025.08.22.671829</dc:identifier>
<dc:title><![CDATA[DTI versus NODDI White Matter Microstructural Biomarkers of Alzheimer's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.24.671810v1?rss=1">
<title>
<![CDATA[
Organoid-evaluable clinical biomarkers predict drug responses and guide new breast cancer therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.24.671810v1?rss=1</link>
<description><![CDATA[
Poor therapeutic response in subsets of breast cancer (BC) patients poses an ongoing challenge. Here, we present a biomarker-guided characterization of 44 patient-derived BC organoids, with the aim of modeling resistant disease with greater fidelity and developing an in-vitro system grounded in clinical data for testing alternative treatment strategies. We utilized patient transcriptomic and outcome data from the I-SPY2 clinical trial to develop predictive models of response to a range of therapies, using only organoid-detectable biomarkers as input. A model predicting response to veliparib-platinum chemotherapy (VP) in triple-negative BC (TNBC) was validated in organoids, showing that in vitro drug responses matched predictions from the patient data-derived model. A drug screen in VP-resistant TNBC organoids identified combination treatments that overcame resistance to cisplatin, including pro-apoptotic therapies. This demonstrates that gene expression-based resistance models derived from patient data can be successfully modeled in organoids that can then be used for therapeutic evaluation.
]]></description>
<dc:creator>Bui, T. B. V.</dc:creator>
<dc:creator>Wolf, D. M.</dc:creator>
<dc:creator>Bruck, M. C.</dc:creator>
<dc:creator>Moore, K.</dc:creator>
<dc:creator>Lien, J.</dc:creator>
<dc:creator>Choi, S. D.</dc:creator>
<dc:creator>Warhadpande, S.</dc:creator>
<dc:creator>Parizadeh, A.</dc:creator>
<dc:creator>Dillon, D.</dc:creator>
<dc:creator>Overmoyer, B.</dc:creator>
<dc:creator>Lynce, F.</dc:creator>
<dc:creator>I-SPY 2 Investigators,</dc:creator>
<dc:creator>Nijman, I. J.</dc:creator>
<dc:creator>Burgering, B. M.</dc:creator>
<dc:creator>Harris, I. S.</dc:creator>
<dc:creator>Esserman, L. J.</dc:creator>
<dc:creator>Van 't Veer, L. J.</dc:creator>
<dc:creator>Rosenbluth, J. M.</dc:creator>
<dc:date>2025-08-28</dc:date>
<dc:identifier>doi:10.1101/2025.08.24.671810</dc:identifier>
<dc:title><![CDATA[Organoid-evaluable clinical biomarkers predict drug responses and guide new breast cancer therapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.26.671755v1?rss=1">
<title>
<![CDATA[
Microtubule Inhibition Triggers MYC-Mediated Immunogenic Cell Death in Breast Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.26.671755v1?rss=1</link>
<description><![CDATA[
Oncogenic MYC promotes cancer cell proliferation, metabolism, and death, while also driving immunosuppression in the tumour microenvironment, complicating immune-based therapies. To counter MYC-driven immune evasion while leveraging MYC-dependent synthetic lethality (MYC-SL), we identified microtubule-targeting agents, including eribulin, as potent inducers of immunogenic cell death in MYChigh triple-negative breast cancer (TNBC). A screen of 528 oncology compounds using damage-associated molecular pattern (DAMP) reporters revealed that microtubule inhibitors induced key DAMPs, including HMGB1 secretion, calreticulin exposure, and double-stranded DNA release, leading to gasdermin-E associated cell death in MYChigh TNBCs. Immune cell co-culture assays showed immune activation, and patient-derived explant cultures confirmed pro-inflammatory cytokine responses. In vivo, cell-free media from eribulin-treated MYChigh murine TNBCs enhanced tumour protection in vaccination models compared to MYC-knockdown controls, linking MYC-dependent DAMP release to immunogenicity. These findings highlight a dual-function therapeutic strategy: agents that selectively induce MYC-dependent immunogenic cell death can provide both targeted cytotoxicity and local immune stimulation, thereby addressing a key limitation of conventional chemotherapeutics, offering a new approach for MYC-driven cancers.
]]></description>
<dc:creator>Aung, J.</dc:creator>
<dc:creator>Liu, R.</dc:creator>
<dc:creator>Peura, A.</dc:creator>
<dc:creator>Hiltunen, A. O.</dc:creator>
<dc:creator>Id, L. J.</dc:creator>
<dc:creator>Savelius, M.</dc:creator>
<dc:creator>Peltonen, M.</dc:creator>
<dc:creator>Turpin, R. J.</dc:creator>
<dc:creator>Salmelin, N.</dc:creator>
<dc:creator>Anttila, J. M.</dc:creator>
<dc:creator>Tervonen, T. A.</dc:creator>
<dc:creator>Suleymanova, I.</dc:creator>
<dc:creator>Nicorici, D.</dc:creator>
<dc:creator>Mutka, M.</dc:creator>
<dc:creator>Mattson, J.</dc:creator>
<dc:creator>Kovanen, P. E.</dc:creator>
<dc:creator>Niinikoski, L.</dc:creator>
<dc:creator>Meretoja, T.</dc:creator>
<dc:creator>Prajapati, B.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:creator>Munne, P.</dc:creator>
<dc:creator>Pouwels, J.</dc:creator>
<dc:creator>Klefstrom, J.</dc:creator>
<dc:date>2025-08-28</dc:date>
<dc:identifier>doi:10.1101/2025.08.26.671755</dc:identifier>
<dc:title><![CDATA[Microtubule Inhibition Triggers MYC-Mediated Immunogenic Cell Death in Breast Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.25.672063v1?rss=1">
<title>
<![CDATA[
Long-term dietary interventions fail to mitigate functional connectivity loss and cognitive decline in the TgF344-AD rat model of Alzheimers Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.25.672063v1?rss=1</link>
<description><![CDATA[
IntroductionAlzheimers disease (AD) is a complex neurodegenerative disorder, characterized by altered brain functional connectivity and network activity, detectable using resting-state functional MRI (rsfMRI). Caloric restriction (CR) and its mimetic resveratrol (Rsv) have shown potential in reducing AD-related pathology and preserving brain function, though research on long-term efficacy is limited. Therefore, we aimed to assess the effects of short- term CR and Rsv administrations on resting-state (rs) functional connectivity (FC), after which we assessed the effects of long-term CR and Rsv interventions on rs-FC, spatial memory, amyloid burden, and neuroinflammation in TgF344-AD rats (Tg) and wild-type (WT) littermates.

Materials and MethodsWe used rsfMRI to investigate rs-FC changes after CR (40%) or daily Rsv supplementation (10 mg/kg, oral) in male and female, WT and TgF344-AD (Tg) rats at the age of 4 months and 11 months, after, respectively, 1 month (short-term) and 8 months (long- term) of either Rsv supplementation, control (Ctrl) or CR diet. Additionally, spatial memory was assessed utilizing the Morris Water Maze (MWM), followed by histological validation of amyloid plaque load (X34), astrogliosis (GFAP) and microgliosis (IBA-1) at 11 months of age.

ResultsBoth long-term CR or Rsv supplementation exert limited effects in the TgF344-AD model on the known AD-related pathological hallmarks. Short-term CR led to a reduction in rs-FC in female TgF344-AD rats compared to Rsv Tg supplemented rats for connections primarily within and between the hippocampal network, while between other connections, CR reduced rs-FC compared to Ctrl in females, irrespective of genotype. Rsv increased rs-FC compared to CR for only a few connections in females, again irrespective of genotype. Long- term CR decreased rs-FC in male Tg CR rats, compared to Tg Ctrl and WT CR rats, primarily for connections within the lateral cortical network (LCN). For other connections, CR reduced rs- FC when compared to Ctrl, irrespective of genotype. Overall, Rsv supplementation showed negligible effects on rs-FC. Moreover, long-term CR yielded modest cognitive improvements in male Tg rats, as evidenced by enhanced performance in the MWM but not in females. Histological validation after long-term dietary intervention revealed region-specific increase or decrease of amyloid burden after CR or Rsv supplementation, respectively. Additionally, CR reduced IBA-1 levels in males, irrespective of genotype, GFAP levels were unaffected by long- term dietary intervention.

ConclusionAltogether, our findings indicate that long-term CR and Rsv exert distinct and limited effects on AD-related pathology in the TgF344-AD model, with CR demonstrating a modest but greater effect on functional and cognitive measures compared to Rsv. This study underscores the difficulty of altering key disease processes with these long-term dietary approaches, highlighting the need for more comprehensive long-term studies to elucidate the potential modulatory role of dietary interventions on AD pathophysiology.
]]></description>
<dc:creator>van Rooij, J. R. A.</dc:creator>
<dc:creator>van den Berg, M.</dc:creator>
<dc:creator>Van Vosselen, M.</dc:creator>
<dc:creator>Calus, E.</dc:creator>
<dc:creator>Vasilkovska, T.</dc:creator>
<dc:creator>Kosten, L.</dc:creator>
<dc:creator>Van Spilbeeck, I.</dc:creator>
<dc:creator>Van Audekerke, J.</dc:creator>
<dc:creator>van Dam, D.</dc:creator>
<dc:creator>Bertoglio, D.</dc:creator>
<dc:creator>Adhikari, M. H.</dc:creator>
<dc:creator>Verhoye, M.</dc:creator>
<dc:date>2025-08-29</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.672063</dc:identifier>
<dc:title><![CDATA[Long-term dietary interventions fail to mitigate functional connectivity loss and cognitive decline in the TgF344-AD rat model of Alzheimers Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.25.672093v1?rss=1">
<title>
<![CDATA[
Angiopoietin signalling is a central axis of amyloid-driven vascular dysfunction in Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.25.672093v1?rss=1</link>
<description><![CDATA[
The neurovascular unit is critical for brain health, and its dysfunction has been linked to Alzheimers disease (AD). However, a cell-type-resolved understanding of how diverse vascular cells become dysfunctional and contribute to disease has been missing. Here, we applied Vessel Isolation and Nuclei Extraction for Sequencing (VINE-seq) to build a comprehensive transcriptomic atlas from 101 individuals along AD progression. Our analysis of over 842,646 parenchymal and vascular nuclei reveals that vascular dysfunction in AD is driven by transcriptional changes rather than shifts in cell proportions, with brain endothelial cells (BECs) and smooth muscle cells (SMCs) most affected. Strikingly, these molecular signatures emerge early at the mild cognitive impairment (MCI) stage, implicating vascular dysfunction early in AD pathogenesis. Stratifying by pathology reveals distinct vascular responses to {beta}-amyloid and tau: {beta}-amyloid burden primarily perturbs BECs and SMCs, while tau pathology predominantly impacts glial cells. We identify dysregulated angiopoietin signaling across multiple vascular cell types as a key axis, with antagonistic ANGPT2 in vascular cells and ANGPT1 in astrocytes becoming progressively dysregulated with AD. Together, this work provides a foundational resource that reveals early and pathology-specific pathways of vascular dysfunction in AD.

Key MessagesO_LIVINE-seq analysis from 101 individuals creates a comprehensive human brain vascular atlas across Alzheimers disease (AD) progression.
C_LIO_LIAD vascular dysfunction is driven by transcriptional changes rather than shifts in cell proportions, with BECs and SMCs most affected.
C_LIO_LITranscriptional signatures of vascular dysfunction emerge early at the mild cognitive impairment (MCI) stage, preceding severe cognitive symptoms and aligning more closely with AD than cognitively normal individuals.
C_LIO_LIA{beta} and tau associate with distinct vascular changes: A{beta} mainly perturbs endothelial and smooth muscle cells, while tau impacts microglia and astrocytes.
C_LIO_LIAngiopoietin signaling (antagonistic ANGPT2 in vascular cells vs. ANGPT1 in astrocytes) becomes progressively dysregulated during AD progression.
C_LI
]]></description>
<dc:creator>Flotho, M.</dc:creator>
<dc:creator>Yang, A.</dc:creator>
<dc:creator>Kern, F.</dc:creator>
<dc:creator>Graf, S.</dc:creator>
<dc:creator>Diks, I. F.</dc:creator>
<dc:creator>Shin, H.</dc:creator>
<dc:creator>Berdnik, D.</dc:creator>
<dc:creator>Agam, M.</dc:creator>
<dc:creator>Channappa, D.</dc:creator>
<dc:creator>Shi, S.</dc:creator>
<dc:creator>Belnap, M. A.</dc:creator>
<dc:creator>Simmons, E.</dc:creator>
<dc:creator>Bradshaw, K.</dc:creator>
<dc:creator>Grandke, F.</dc:creator>
<dc:creator>Bennett, D. A.</dc:creator>
<dc:creator>Buckwalter, M.</dc:creator>
<dc:creator>Keller, A.</dc:creator>
<dc:creator>Wyss-Coray, T.</dc:creator>
<dc:date>2025-08-29</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.672093</dc:identifier>
<dc:title><![CDATA[Angiopoietin signalling is a central axis of amyloid-driven vascular dysfunction in Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.27.672733v1?rss=1">
<title>
<![CDATA[
iPSC-Derived Hepatocytes from Patients with MASLD Exhibit Early Mitochondrial Dysfunction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.27.672733v1?rss=1</link>
<description><![CDATA[
Background & AimsA hallmark of metabolic dysfunction-associated steatotic liver disease (MASLD) is a decline in the ability of hepatocyte mitochondria to adapt to excess lipid. This leads to the production of reactive oxygen species (ROS) and the instigation of a vicious cycle of further mitochondrial damage and cellular dysfunction that promotes disease progression. In this study, we investigated whether induced pluripotent stem cells (iPSCs) from MASLD patients exhibit features of mitochondrial dysfunction when differentiated to hepatocyte-like cells (iPSC-Heps).

MethodsiPSCs from 10 MASLD patients and 10 healthy control subjects genotyped for the I148M variant of PNPLA3 were differentiated to iPSC-Heps. Mitochondrial mass and function were assessed under basal culture conditions and following short-term exposure to exogenous palmitate. Outcomes included gene expression, mitochondrial oxygen consumption, ROS production and cellular energy status.

ResultsiPSC-Heps from MASLD patients spontaneously accrued more lipid than control iPSC-Heps. Mitochondrial content was similar in MASLD and control iPSC-Heps, but MASLD iPSC-Heps displayed significant differences in mitochondrial function including a decrease in oxygen consumption rate when challenged with palmitate. Antioxidant gene expression was increased at baseline in MASLD vs. control iPSC-Heps, and MASLD iPSC-Heps produced more ROS and less ATP than controls after palmitate treatment. Differences persisted even when controlling for PNPLA3 genotype.

ConclusionsiPSC-Heps from MASLD patients exhibit mitochondrial alterations characteristic of their diseased origin. The degree of mitochondrial dysfunction seen in MASLD iPSC-Heps is reminiscent of that described clinically in early MASLD, prior to progression to steatohepatitis. Mitochondrial alterations in MASLD iPSC-Heps occur independently of PNPLA3 genotype.
]]></description>
<dc:creator>Le Guillou, D.</dc:creator>
<dc:creator>Siao, K.</dc:creator>
<dc:creator>Her, C. L.</dc:creator>
<dc:creator>Duwaerts, C. C.</dc:creator>
<dc:creator>Maher, J. J.</dc:creator>
<dc:date>2025-08-30</dc:date>
<dc:identifier>doi:10.1101/2025.08.27.672733</dc:identifier>
<dc:title><![CDATA[iPSC-Derived Hepatocytes from Patients with MASLD Exhibit Early Mitochondrial Dysfunction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.26.672349v1?rss=1">
<title>
<![CDATA[
Dynamic neural states underpin bradykinesia severity in Parkinsons disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.26.672349v1?rss=1</link>
<description><![CDATA[
BackgroundBradykinesia in Parkinsons disease (PD) may arise due to transient, network-wide neural dynamics that extend beyond beta-band oscillatory activity within the motor cortical-subthalamic nucleus (STN) circuit.

MethodsWe address this question by using Hidden Markov Models (HMMs) to identify neural states from chronic motor cortical and STN recordings in five PD patients (1,046 hours from 10 hemispheres), with concurrent measurements of bradykinesia using wearable sensors.

FindingsWe identified four neural states with distinct spectral and temporal features. Two states exhibited spectral signatures--particularly STN low and high gamma, STN delta/alpha, cortical beta, and cortico-STN beta coherence--that predicted worsening bradykinesia. However, STN beta power alone was not consistently predictive, challenging traditional beta-centric views. These states also displayed compensatory features associated with bradykinesia amelioration, including cortical delta/alpha activity, cortical high gamma, and cortico-STN high gamma coherence. Two additional states affected bradykinesia through temporal rather than spectral properties. Prolonged lifetimes of one of these state worsened symptoms, whereas increased occurrences of another, marked by local beta without cortico-STN beta coherence, improved motor function.

InterpretationOur findings highlight the multidimensional nature of bradykinesia and suggest that state-aware, adaptive interventions targeting state features--rather than single frequency bands--may offer new opportunities for improved deep brain stimulation in PD.

FundingAO is supported by an MRC Clinician Scientist Fellowship (MR/W024810/1) and a Rosetrees Trust/Race Against Dementia Team award. BA and AO acknowledge funding support from the Oxford University Hospitals Charity and Jon Moulton Trust. TL acknowledges funding support from the China Scholarship Council.

Research in ContextO_ST_ABSEvidence before this studyC_ST_ABSPrevious work has demonstrated that subthalamic nucleus oscillatory activity at beta (15-30 Hz) frequencies correlates positively with motor symptoms in Parkinsons disease. This has led to beta activity being used as a biomarker for adaptive Deep Brain Stimulation. It remains unclear however whether other oscillatory features within the broader motor cortical-subthalamic nucleus network could provide improved biomarkers for tracking symptom severity.

Added value of this studyWe address this by performing chronic motor cortical and subthalamic nucleus recordings in Parkinsons disease patients during activities of daily living. Simultaneous measurements of symptom severity were captured using wearable sensors. We used Hidden Markov Models to identify transient states of neural activity and related these to symptom severity. Although cortical beta and cortico-STN beta coherence predicted worsening motor symptoms, STN beta activity alone was not a consistent predictor. Interestingly, we identified spectral features associated with motor symptom improvements, including cortical delta/alpha activity, cortical high gamma, and cortico-STN high gamma coherence. Additionally, there was a  compensatory state characterised by short-lived cortical and subthalamic nucleus beta activity, whose increased occurrence was associated with symptomatic improvements.

Implications of all the available evidenceOur findings highlight the importance of prolonged, high temporal resolution measurements of both neural activity and symptom severity for discovering adaptive Deep Brain Stimulation biomarkers. Crucially, we identify new target states and spectral features for improving motor symptoms in Parkinsons disease.
]]></description>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Sargezeh, B. A.</dc:creator>
<dc:creator>Hahn, A.</dc:creator>
<dc:creator>Shcherbakova, M.</dc:creator>
<dc:creator>Neumann, W. J.</dc:creator>
<dc:creator>Little, S.</dc:creator>
<dc:creator>Starr, P.</dc:creator>
<dc:creator>Oswal, A.</dc:creator>
<dc:date>2025-08-31</dc:date>
<dc:identifier>doi:10.1101/2025.08.26.672349</dc:identifier>
<dc:title><![CDATA[Dynamic neural states underpin bradykinesia severity in Parkinsons disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.28.672801v1?rss=1">
<title>
<![CDATA[
Cryptic splicing in synaptic and membrane excitability genes links TDP-43 loss to neuronal dysfunction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.28.672801v1?rss=1</link>
<description><![CDATA[
TDP-43 pathology is a defining pathological hallmark of multiple neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). A major feature of TDP-43 pathology is its nuclear depletion, leading to the aberrant inclusion of cryptic exons during RNA splicing. STMN2 and UNC13A have emerged as prominent TDP-43 splicing targets, but the broader impact of TDP-43-dependent cryptic splicing on neuronal function remains unclear. Here, we report new TDP-43 splicing targets critical for membrane excitability and synaptic function, including KALRN, RAP1GAP, SYT7 and KCNQ2. Using human stem cell-derived neurons, we show that TDP-43 reduction induces cryptic splicing and downregulation of these genes, resulting in impaired excitability and synaptic transmission. In postmortem brains from patients with FTD, these cryptic splicing events occur selectively in neurons with TDP-43 pathology. Importantly, suppressing individual cryptic splicing events using antisense oligonucleotides partially restores neuronal function, and combined targeting almost fully rescues the synaptic deficit caused by TDP-43 loss. Together, our findings provide evidence that cryptic splicing in these synaptic and membrane excitability genes is not only a downstream marker but instead a direct driver of neuronal dysfunction, establishing a mechanistic link between TDP-43 pathology and neurodegeneration in ALS and FTD.
]]></description>
<dc:creator>Guo, C.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Vatsavayai, S. C.</dc:creator>
<dc:creator>Aikyama, T.</dc:creator>
<dc:creator>Zeng, Y.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Sianto, O.</dc:creator>
<dc:creator>Yang, E.</dc:creator>
<dc:creator>Bombosch, J.</dc:creator>
<dc:creator>Powell, R.</dc:creator>
<dc:creator>Zhen, S.</dc:creator>
<dc:creator>Mekhoubad, S.</dc:creator>
<dc:creator>Morrie, R. D.</dc:creator>
<dc:creator>Miller, G.</dc:creator>
<dc:creator>Green, E. M.</dc:creator>
<dc:creator>Petrucelli, L.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Gitler, A. D.</dc:creator>
<dc:date>2025-09-02</dc:date>
<dc:identifier>doi:10.1101/2025.08.28.672801</dc:identifier>
<dc:title><![CDATA[Cryptic splicing in synaptic and membrane excitability genes links TDP-43 loss to neuronal dysfunction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.29.673168v1?rss=1">
<title>
<![CDATA[
Atypical developmental remodeling of dopamine neurons involves AKT-GSK3β signaling and glia-mediated axon degeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.29.673168v1?rss=1</link>
<description><![CDATA[
Neuronal remodeling is essential for sculpting neural circuits, and its disruption has been implicated in neurodevelopmental and neuropsychiatric disorders. Yet the molecular and cellular diversity of remodeling across neuron types remains incompletely understood. Here, we uncover a distinct remodeling mode in a subtype of Drosophila dopamine neurons (DANs) critical for learning, memory, sleep, and locomotion. Unlike the stereotypical pruning-then-regrowth paradigm, these DANs undergo a transient axon overgrowth followed by selective pruning during metamorphosis. Remarkably, DAN axon pruning proceeds independently of canonical ecdysone signaling and instead involves neuron-intrinsic AKT-GSK3{beta} signaling and extrinsic glial activity. Disruption of AKT-GSK3{beta} signaling alters microtubule stability and impairs glial recruitment and clearance of axonal debris. Notably, the role of AKT-GSK3{beta} is cell-type specific, underscoring mechanistic diversity in remodeling programs. These findings reveal an unexpected overgrowth-then-pruning developmental trajectory, establishing DANs as a powerful model to uncover the mechanisms underlying neuronal remodeling, circuit maturation, and neurodegeneration.
]]></description>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Tu, X.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Davis, R. L.</dc:creator>
<dc:creator>Jan, L. Y.</dc:creator>
<dc:creator>Jan, Y. N.</dc:creator>
<dc:date>2025-09-03</dc:date>
<dc:identifier>doi:10.1101/2025.08.29.673168</dc:identifier>
<dc:title><![CDATA[Atypical developmental remodeling of dopamine neurons involves AKT-GSK3β signaling and glia-mediated axon degeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.30.673205v1?rss=1">
<title>
<![CDATA[
Understanding and imaging PHGDH-driven intrinsic resistance to mutant IDH inhibition in gliomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.30.673205v1?rss=1</link>
<description><![CDATA[
Mutations in isocitrate dehydrogenase (IDHm) define a distinct molecular class of gliomas. IDHm converts -ketoglutarate (-KG) to the oncometabolite D-2-hydroxyglutarate (D-2HG), which drives tumorigenesis. The IDHm inhibitor vorasidenib suppresses D-2HG production and extends progression-free survival in some, but not all, IDHm glioma patients. Here, using clinically relevant patient-derived IDHm models and patient tissue, we show that phosphoglycerate dehydrogenase (PHGDH) drives intrinsic resistance to vorasidenib by promiscuously converting -KG to D-2HG and maintaining D-2HG concentration despite IDHm inhibition. Silencing PHGDH sensitizes resistant models to vorasidenib, while conversely, overexpressing PHGDH induces vorasidenib resistance in sensitive models. Importantly, deuterium metabolic imaging of D-2HG production from diethyl-[3,3-2H]--ketoglutarate provides an early readout of response and resistance to vorasidenib that is not available by anatomical imaging in vivo. Collectively, we have identified PHGDH-driven D-2HG production as an intrinsic mechanism of resistance to vorasidenib and diethyl-[3,3-2H]-ketoglutarate as a non-invasive tracer for interrogating intrinsic resistance in IDHm gliomas.

STATEMENT OF SIGNIFICANCEVorasidenib, which suppresses D-2HG production, is the first precision therapy to be approved for IDHm glioma patients. We show that PHGDH-driven restoration of D-2HG production mediates intrinsic resistance to vorasidenib in IDHm gliomas. Importantly, deuterium metabolic imaging of D-2HG production from diethyl-[3,3-2H]--ketoglutarate enables non-invasive assessment of resistance in IDHm gliomas.
]]></description>
<dc:creator>Taglang, C.</dc:creator>
<dc:creator>Batsios, G.</dc:creator>
<dc:creator>Gillespie, A. M.</dc:creator>
<dc:creator>Phillips, J. J.</dc:creator>
<dc:creator>Taylor, J. W.</dc:creator>
<dc:creator>Viswanath, P.</dc:creator>
<dc:date>2025-09-04</dc:date>
<dc:identifier>doi:10.1101/2025.08.30.673205</dc:identifier>
<dc:title><![CDATA[Understanding and imaging PHGDH-driven intrinsic resistance to mutant IDH inhibition in gliomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.02.673643v1?rss=1">
<title>
<![CDATA[
Spatiotemporal cellular dynamics of the notochord shape intervertebral disc morphogenesis in the mouse embryo through apoptosis and proliferation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.02.673643v1?rss=1</link>
<description><![CDATA[
BackgroundThe notochord is a midline structure essential for vertebrate embryogenesis, contributing to the development of the nervous system, digestive tract, and vertebral column. In particular, notochord signaling is indispensable for proper patterning and coordinated development of alternating vertebrae and intervertebral discs (IVDs). Later, notochordal cells (NCs) mature and adopt a characteristic vacuolated morphology before giving rise to the core of the forming IVD, the nucleus pulposus (NP). Postnatally, NCs play pivotal role in maintaining disc integrity through the secretion of specific factors and extracellular matrix (ECM). Despite its importance in disc formation and homeostasis, the morphogenetic mechanisms underlying the notochords transformation into the NP are insufficiently characterized.

ResultsWe conducted a comprehensive histological and immunohistochemical analysis to investigate the cellular events governing NP formation in the mouse developing spine. Temporal analysis of intracytoplasmic vacuole formation using Lamp1 marker revealed their contribution to NP growth, while cell density progressively decreased. In addition, quantitative analyses demonstrated a notable proliferative capacity within notochordal cells coupled with region-specific apoptotic activity in the sclerotome, at future disc sites.

ConclusionsThis study highlights the intricate balance of cellular proliferation, programmed cell death, matrix remodeling, and vacuolation dynamics as key determinants in shaping the NP along the rostro-caudal axis.

Key Findings- Spatiotemporal cellular changes drive the transition from notochord to nucleus pulposus
- Future disc regions show selective notochord proliferation and sclerotome cell death
- Notochord vacuolization and matrix deposition contribute to nucleus pulposus morphogenesis
]]></description>
<dc:creator>Warin, J.</dc:creator>
<dc:creator>Balissat, E.</dc:creator>
<dc:creator>Colombier, P.</dc:creator>
<dc:creator>Paillat, L.</dc:creator>
<dc:creator>Dutilleul, M.</dc:creator>
<dc:creator>Camus, A.</dc:creator>
<dc:date>2025-09-04</dc:date>
<dc:identifier>doi:10.1101/2025.09.02.673643</dc:identifier>
<dc:title><![CDATA[Spatiotemporal cellular dynamics of the notochord shape intervertebral disc morphogenesis in the mouse embryo through apoptosis and proliferation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.01.673567v1?rss=1">
<title>
<![CDATA[
Spatial Single-Cell Atlas Reveals KSHV-Driven Broad Cellular Reprogramming, Progenitor Expansion, Immune and Vascular Remodeling in Kaposi's Sarcoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.01.673567v1?rss=1</link>
<description><![CDATA[
Kaposis sarcoma (KS) is a highly inflammatory, angiogenic tumor driven by Kaposis sarcoma-associated herpesvirus (KSHV), yet the origins of tumor cells and mechanisms of progression remain unclear. Here, we present the first spatial single-cell atlas of KS, profiling 256 samples across patch, plaque, and nodular lesions and normal controls. We identify CD34 progenitor lymphatic endothelial cells (LECs) as the primary targets of KSHV, whose clonal expansion drives tumor growth. KSHV infection induces widespread cellular reprogramming across the tumor microenvironment, including LECs, vascular endothelial cells, fibroblasts, and macrophages, generating hybrid phenotypes that support angiogenesis, inflammation, and immune modulation. KSHV macrophages are enriched in tumor-proximal niches, further promoting a proangiogenic, immunosuppressive environment. Spatial analysis reveals evolving tumor-associated niches, with a core-to-periphery gradient correlating with infection, immune modulation, and cellular remodeling. We identify disease progression predictive signatures, offering mechanistic insights into KS pathogenesis and potential new therapeutic strategies by reprogramming the tumor microenvironment.

Highlights CD34+ progenitor lymphatic endothelial cells are the primary KSHV target cells, with their clonal expansion driving Kaposis sarcoma growth.
 KSHV infection reprograms broad cell types into hybrid identities, and drives tumor-specific niche and vascular remodeling, endothelial plasticity, and immune modulation.
 KSHV-reprogrammed macrophages drive inflammation, angiogenesis, and immune modulation.
 Spatially resolved molecular and cellular signatures predict Kaposis sarcoma progression, offering novel therapeutic strategies targeting the tumor microenvironment.
]]></description>
<dc:creator>Meng, W.</dc:creator>
<dc:creator>Das, A.</dc:creator>
<dc:creator>Sinha, H.</dc:creator>
<dc:creator>Naous, R.</dc:creator>
<dc:creator>Bracci, P.</dc:creator>
<dc:creator>McGrath, M.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Gao, S.-J.</dc:creator>
<dc:date>2025-09-05</dc:date>
<dc:identifier>doi:10.1101/2025.09.01.673567</dc:identifier>
<dc:title><![CDATA[Spatial Single-Cell Atlas Reveals KSHV-Driven Broad Cellular Reprogramming, Progenitor Expansion, Immune and Vascular Remodeling in Kaposi's Sarcoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.03.673593v1?rss=1">
<title>
<![CDATA[
Incubator-Free Organoid Culture in a Sealed Recirculatory System 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.03.673593v1?rss=1</link>
<description><![CDATA[
Discovery in human biology is pivoting toward high-dimensional computational analysis of 3D in vitro models, but this progress is limited by reliance on conventional cell culture techniques. Realism and data collection are hindered by the environmental instabilities and accessibility constraints of standard incubators. We introduce an automated, sealed recirculatory system that eliminates these barriers, enabling unconstrained instrument integration and infrastructure-independent scalability. By employing gas-tight sealing, a liquid-phase gas buffer and a non-porous plastic gas exchanger, our technology maintains biological stability without the compromises of open-air vessels. This design eliminates the need for CO2 incubators and prevents the evaporative drift that typically plagues conventional open-culture vessels. Operating on the benchtop outside the cell culture suite, we demonstrate that our system supports continuous, multi-week live imaging of vascular organoids while maintaining metabolic viability, structural fidelity and electrophysiological activity in brain organoids comparable to traditional in-incubator cultures.

TeaserA sealed benchtop device enables data-rich organoid culture beyond the constraints of the cell culture laboratory.
]]></description>
<dc:creator>Rosen, Y.</dc:creator>
<dc:creator>Doganyigit, K.</dc:creator>
<dc:creator>Arul, S.</dc:creator>
<dc:creator>Wachtel, E.</dc:creator>
<dc:creator>Ehrlich, D.</dc:creator>
<dc:creator>Venuturimilli, V.</dc:creator>
<dc:creator>Mouzaya, M.</dc:creator>
<dc:creator>Oriaku, S.</dc:creator>
<dc:creator>Ndiforchu, B.</dc:creator>
<dc:creator>Kaurala, G.</dc:creator>
<dc:creator>Hernandez, S.</dc:creator>
<dc:creator>Moarefian, M.</dc:creator>
<dc:creator>Schweiger, H.</dc:creator>
<dc:creator>Cisneros, A.</dc:creator>
<dc:creator>Zeraatkar, M.</dc:creator>
<dc:creator>Geng, J.</dc:creator>
<dc:creator>Mostajo-Radji, M. A.</dc:creator>
<dc:creator>Winkler, E.</dc:creator>
<dc:creator>Haussler, D.</dc:creator>
<dc:creator>Teodorescu, M.</dc:creator>
<dc:date>2025-09-05</dc:date>
<dc:identifier>doi:10.1101/2025.09.03.673593</dc:identifier>
<dc:title><![CDATA[Incubator-Free Organoid Culture in a Sealed Recirculatory System]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.04.674293v1?rss=1">
<title>
<![CDATA[
Sex differences in the developing human cortex intersect with genetic risk of neurodevelopmental disorders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.04.674293v1?rss=1</link>
<description><![CDATA[
Autism is highly heritable and diagnosed more frequently in males than females. To identify neurodevelopmental processes that might present sex-biased vulnerability, we generated transcriptomic and epigenomic profiles of cell types present in the prenatally developing human cerebral cortex of 27 males and 21 females. By intersecting sex-biased molecular signatures and genes with de novo mutations in male and female autistic probands, we reveal two points of vulnerability contributing to the sex-biased penetrance in neurodevelopmental disorders (NDDs). First, we show that NDD risk genes are biased towards higher expression in females, identifying the NDD gene MEF2C as a critical transcription factor for female-biased expression. Second, we identify a significant contribution of X chromosome genes to NDD pathobiology. We construct a gene regulatory map of X-linked risk genes to enable functional studies of genetic variants that likely disrupt gene expression in the developing brains of autistic males. Together, these results point towards an outsized contribution of the X-chromosome to both the origin of sex differences in the developing human cortex and NDD vulnerability. We propose a model where female-biased vulnerability is driven by coding variation within genes while male-biased vulnerability is driven by noncoding variation in regulatory elements that affect gene expression.
]]></description>
<dc:creator>Hennick, K.</dc:creator>
<dc:creator>Sui, Y.</dc:creator>
<dc:creator>Karunakaran, D.</dc:creator>
<dc:creator>Nicolella, A.</dc:creator>
<dc:creator>Leonard, R.</dc:creator>
<dc:creator>Meyer-Schuman, R.</dc:creator>
<dc:creator>Berk-Rauch, H.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Chakravarti, A.</dc:creator>
<dc:creator>Zoghbi, H. Y.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:date>2025-09-05</dc:date>
<dc:identifier>doi:10.1101/2025.09.04.674293</dc:identifier>
<dc:title><![CDATA[Sex differences in the developing human cortex intersect with genetic risk of neurodevelopmental disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.05.674553v1?rss=1">
<title>
<![CDATA[
MitoScribe single-cell molecular recorder logs graded signaling dynamics into mitochondrial DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.05.674553v1?rss=1</link>
<description><![CDATA[
Genetically encoded DNA recorders convert transient biological events into stable genomic mutations, offering a means to reconstruct past cellular states. However, current approaches to log historical events by modifying genomic DNA have limited capacity to record the magnitude of biological signals within individual cells. Here, we introduce MitoScribe, a mitochondrial DNA (mtDNA)-based recording platform that uses mtDNA base editors (DdCBEs) to write graded biological signals into mtDNA as neutral, single-nucleotide substitutions at a defined site. Taking advantage of the hundreds to thousands of mitochondrial genome copies per cell, we demonstrate MitoScribe enables reproducible, highly sensitive, non-destructive, durable, and high-throughput measurements of molecular signals, including hypoxia, NF-{kappa}B activity, BMP and Wnt signaling. We show multiple modes of operation, including multiplexed recordings of two independent signals, and coincidence detection of temporally overlapping signals. Coupling MitoScribe with single-cell RNA sequencing and mitochondrial transcript enrichment, we further reconstruct signaling dynamics at the single-cell transcriptome level. Applying this approach during the directed differentiation of human induced pluripotent stem cells (iPSCs) toward mesoderm, we show that early heterogeneity in response to a differentiation cue predicts the later cell state. Together, MitoScribe provides a scalable platform for high-resolution molecular recording in complex cellular contexts.
]]></description>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Poulis, N.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Shipman, S.</dc:creator>
<dc:date>2025-09-05</dc:date>
<dc:identifier>doi:10.1101/2025.09.05.674553</dc:identifier>
<dc:title><![CDATA[MitoScribe single-cell molecular recorder logs graded signaling dynamics into mitochondrial DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.02.673780v1?rss=1">
<title>
<![CDATA[
BOGO: A Proteome-Wide Gene Overexpression Platform for Discovering Rational Cancer Combination Therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.02.673780v1?rss=1</link>
<description><![CDATA[
O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/673780v2_ufig1.gif" ALT="Figure 1">
View larger version (56K):
org.highwire.dtl.DTLVardef@19d5d7dorg.highwire.dtl.DTLVardef@64f7eborg.highwire.dtl.DTLVardef@d08b75org.highwire.dtl.DTLVardef@173c448_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOGraphical AbstractC_FLOATNO C_FIG Cancer drug resistance remains a major barrier to durable treatment success, often leading to relapse despite advances in precision oncology. While combination therapies are being increasingly investigated, such as chemotherapy with small molecule inhibitors, predicting drug response and identifying rational drug combinations based on resistance mechanisms remain major challenges. Therefore, a proteome-wide, single-gene overexpression screening platform is essential for guiding rational therapy selection. Here, we present BOGO (Bxb1-landing pad human ORFeome-integrated system for a proteome-wide Gene Overexpression), a robust, scalable, and reproducible screening platform that enables single-copy, site-specific integration and overexpression of [~]19,000 human open across cancer cell models. Using BOGO, we identified drug-specific response drivers for 16 chemotherapeutic agents and integrated clinical datasets to uncover proliferation and resistance-associated genes with prognostic potential. Drug response similarity networks revealed both shared and unique mechanisms, highlighting key pathways such as autophagy, apoptosis, and Wnt signaling, and notable resistance-associated genes including BCL2, POLD2, and TRADD. In particular, we proposed a synergistic combination of the BCL2 family inhibitor ABT-263 (Navitoclax(R)) and the DNA analog TAS-102 (Lonsurf(R)), which revealed that lysosomal modulation is a key mechanism driving DNA analog resistance. This combination therapy selectively enhanced cytotoxicity in colorectal and pancreatic cancer cells in vitro, and demonstrated therapeutic benefit in vivo in both cell line-derived xenograft (CDX) and patient-derived xenograft (PDX) models. Together, these findings establish BOGO as a powerful gene overexpression perturbation platform for systematically identifying chemoresistance and chemosensitization drivers, and for discovering rational combination therapies. Its scalability and reproducibility position BOGO as a broadly applicable tool for functional genomics and therapeutic discovery beyond cancer resistance.
]]></description>
<dc:creator>Jo, K. B.</dc:creator>
<dc:creator>Alruwaili, M. M.</dc:creator>
<dc:creator>Kim, D.-E.</dc:creator>
<dc:creator>Koh, Y.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>You, K.</dc:creator>
<dc:creator>Kim, J.-S.</dc:creator>
<dc:creator>Sane, S.</dc:creator>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Wright, J. P.</dc:creator>
<dc:creator>Lim, H. J.</dc:creator>
<dc:creator>Naranjo, M. N.</dc:creator>
<dc:creator>Cote, A. G.</dc:creator>
<dc:creator>Roth, F. P.</dc:creator>
<dc:creator>Hill, D. E.</dc:creator>
<dc:creator>Choi, J.-H.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Matreyek, K. A.</dc:creator>
<dc:creator>Farh, K. K.- H.</dc:creator>
<dc:creator>Park, J.-E.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Bakin, A. V.</dc:creator>
<dc:creator>Kim, D.-K.</dc:creator>
<dc:date>2025-09-07</dc:date>
<dc:identifier>doi:10.1101/2025.09.02.673780</dc:identifier>
<dc:title><![CDATA[BOGO: A Proteome-Wide Gene Overexpression Platform for Discovering Rational Cancer Combination Therapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.05.674489v1?rss=1">
<title>
<![CDATA[
Targeting p16INK4a reverses alveolar epithelial cell dysfunction and induces lung regeneration in emphysema 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.05.674489v1?rss=1</link>
<description><![CDATA[
Pulmonary emphysema involves impaired regenerative capacity of alveolar type 2 epithelial cells (AT2), the main progenitor cells in alveoli. However, the mechanisms underlying dysfunctional epithelial repair remain unclear. In a mouse model of elastase-induced emphysema, we observed an accumulation of activated AT2s in the lung, associated with an overexpression of p16INK4a (p16), a cell cycle inhibitor known to influence stem cell fate. Deletion of p16 promoted the transition of AT2 into alveolar type 1 (AT1) cells, resulting in tissue regeneration in both mice and alveolar organoids. Pharmacological targeting of the p16 pathway using senolytic agents recapitulate this regenerative effect, further supporting the role of p16 as a key brake on epithelial plasticity. These findings demonstrate that alveolar epithelial cell dysfunction can be reversed by p16 deletion or by eliminating p16+ cells, thereby reactivating the AT2-to-AT1 transition and promoting endogenous alveolar regeneration. This work identifies the p16 pathway as a promising therapeutic target for restoring damaged alveoli in emphysema.
]]></description>
<dc:creator>Ribeiro Baptista, B.</dc:creator>
<dc:creator>Toigo, M.</dc:creator>
<dc:creator>Justeau, G.</dc:creator>
<dc:creator>Abou-atmeh, P.</dc:creator>
<dc:creator>De Freitas Castro, T.</dc:creator>
<dc:creator>Zysman, M.</dc:creator>
<dc:creator>Thiebaut De Menonville, C.</dc:creator>
<dc:creator>Audureau, E.</dc:creator>
<dc:creator>Schnyder, K.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Koenigshoff, M.</dc:creator>
<dc:creator>Lanone, S.</dc:creator>
<dc:creator>Chabot, F.</dc:creator>
<dc:creator>Zana-Taieb, E.</dc:creator>
<dc:creator>Jourdan Le Saux, C.</dc:creator>
<dc:creator>Lehmann, M.</dc:creator>
<dc:creator>Derumeaux, G.</dc:creator>
<dc:creator>Boczkowski, J.</dc:creator>
<dc:creator>Gote-Schniering, J.</dc:creator>
<dc:creator>Boyer, L.</dc:creator>
<dc:date>2025-09-07</dc:date>
<dc:identifier>doi:10.1101/2025.09.05.674489</dc:identifier>
<dc:title><![CDATA[Targeting p16INK4a reverses alveolar epithelial cell dysfunction and induces lung regeneration in emphysema]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.08.674823v1?rss=1">
<title>
<![CDATA[
De novo design of semisynthetic protein nanopores 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.08.674823v1?rss=1</link>
<description><![CDATA[
Protein nanopores are essential components of single-molecule oligonucleotide sequencing and sensing devices. Here, we demonstrate that installing additional de novo subunits enables large-scale architectural changes of nanopore complexes. We design de novo proteins that integrate seamlessly with the CsgG pore to form 18-subunit, 315-kilodalton complexes with precisely sculpted pore architectures and tailored ion conduction, opening new possibilities for engineering enhanced nanopores with customized structural and functional properties.
]]></description>
<dc:creator>Schnaider, L.</dc:creator>
<dc:creator>Hatstat, A. K.</dc:creator>
<dc:creator>Scott, A. J.</dc:creator>
<dc:creator>Tan, S. K.</dc:creator>
<dc:creator>Hambley, R. G.</dc:creator>
<dc:creator>Dawson, W. M.</dc:creator>
<dc:creator>Griffiths, R. C.</dc:creator>
<dc:creator>Kormos, R. C.</dc:creator>
<dc:creator>Melo, A. A.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Polizzi, N. C.</dc:creator>
<dc:creator>Wallace, E. J.</dc:creator>
<dc:creator>Merz, G. E.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:date>2025-09-08</dc:date>
<dc:identifier>doi:10.1101/2025.09.08.674823</dc:identifier>
<dc:title><![CDATA[De novo design of semisynthetic protein nanopores]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.06.674665v1?rss=1">
<title>
<![CDATA[
Conserved lipid metabolic reprogramming confers hypoxic and aging resilience 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.06.674665v1?rss=1</link>
<description><![CDATA[
The Arctic ground squirrel (AGS, Urocitellus parryii), an extreme hibernator, exhibits remarkable resilience to stressors like hypoxia and hypothermia, making it an ideal model for studying cellular metabolic adaptation. The underlying mechanisms of AGS resilience are largely unknown. Here, we use lipidomic and metabolomic profiling to discover specific downregulation of triglyceride lipids and upregulation of the lipid biosynthetic precursor malonic acid in AGS neural stem cells (NSC) versus murine NSCs. Inhibiting lipid biosynthesis recapitulates hypoxic resilience of squirrel NSCs. Extending this model, we find that acute exposure to hypoxia downregulates key lipid biosynthetic enzymes in C. elegans, while inhibiting lipid biosynthesis reduces mitochondrial fission and facilitates hypoxic survival. Moreover, inhibiting lipid biosynthesis protects against APOE4-induced pathologies and aging trajectories in C. elegans. These findings suggest triglyceride downregulation as a conserved metabolic resilience mechanism, offering insights into protective strategies for neural tissues under hypoxic or ischemic conditions, APOE4-induced pathologies and aging.
]]></description>
<dc:creator>Jiang, W. I.</dc:creator>
<dc:creator>Vale, G. D. d.</dc:creator>
<dc:creator>Pearce, Q.</dc:creator>
<dc:creator>Kong, K.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>McDonald, J. G.</dc:creator>
<dc:creator>Cox, J. E.</dc:creator>
<dc:creator>Singhal, N. S.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2025-09-09</dc:date>
<dc:identifier>doi:10.1101/2025.09.06.674665</dc:identifier>
<dc:title><![CDATA[Conserved lipid metabolic reprogramming confers hypoxic and aging resilience]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.08.674963v1?rss=1">
<title>
<![CDATA[
Redox regulation of neuroinflammatory pathways contributes to damage in Alzheimers disease brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.08.674963v1?rss=1</link>
<description><![CDATA[
The mechanism(s) whereby redox stress mediates aberrant immune signaling in age-related neurological disorders remains largely unknown. Normally, the innate immune system mounts a robust response to infectious stimuli. However, unintentional activation by host-derived factors, such as aggregated proteins associated with neurodegenerative disorders or by cytoplasmic genomic or mitochondrial DNA, can elicit aberrant immune responses. One such immune response is represented by the cytosolic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway. Using redox chemical biology and mass spectrometry approaches, we identified S-nitrosylation of STING cysteine 148 as a novel posttranslational redox modification underlying aberrant type 1 interferon signaling in Alzheimers disease (AD). Critically, we observed S-nitrosylated STING (SNO-STING) in postmortem human AD brains, in hiPSC-derived microglia (hiMG) exposed to amyloid-{beta} (A{beta})/-synuclein (Syn) aggregates, and in 5xFAD transgenic mice. Mechanistically, our findings reveal that STING S-nitrosylation is critical in initiating signaling cascades by promoting the formation of disulfide-bonded STING oligomers. This leads to neuroinflammation early in the course of disease in vivo in 5xFAD mice with consequent synaptic loss. Collectively, our research supports the role of SNO-STING in neuroinflammation associated with AD, and points to a novel druggable cysteine residue in STING to prevent this S-nitrosylation reaction with its inherent inflammatory response.

One Sentence SummaryS-Nitrosylation of STING triggers activation of cGAS-STING signaling in Alzheimers disease brain and subserves a novel link between excessive nitrosative stress and dysregulated innate immunity, thus contributing to disease progression.
]]></description>
<dc:creator>Carnevale, L. N.</dc:creator>
<dc:creator>Banerjee, P.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Navarro, J.</dc:creator>
<dc:creator>Raspur, C. K.</dc:creator>
<dc:creator>Nakamura, T.</dc:creator>
<dc:creator>Schahrer, E.</dc:creator>
<dc:creator>Scott, H.</dc:creator>
<dc:creator>Lang, N.</dc:creator>
<dc:creator>Diedrich, J. K.</dc:creator>
<dc:creator>Yates, J. R.</dc:creator>
<dc:creator>Lipton, S. A.</dc:creator>
<dc:date>2025-09-09</dc:date>
<dc:identifier>doi:10.1101/2025.09.08.674963</dc:identifier>
<dc:title><![CDATA[Redox regulation of neuroinflammatory pathways contributes to damage in Alzheimers disease brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.10.675278v1?rss=1">
<title>
<![CDATA[
The role of intrinsically disordered domains in regulating G protein coupled receptor signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.10.675278v1?rss=1</link>
<description><![CDATA[
The 2A adrenergic receptor (2AAR) is a clinically important target for various diseases including hypertension, diabetes and chronic pain. Here, using single-molecule fluorescence resonance energy transfer imaging, we show how agonist-specific activation dynamics in both structured transmembrane domain (TMD) and intrinsically disorders regions (IDRs) of 2AAR lead to diverse signaling profiles. Through seven pairs of strategically designed fluorophore labels, we systematically investigate the real-time conformational changes of 2AAR. Our study reveals unique TM6 dynamics in 2AAR, featured by a high energy barrier for agonist-induced outward movements essential for activation. In contrast, we identify agonist-specific conformational dynamics of a partially disordered extracellular loop (ECL2), highlighting its role as a dynamic regulatory module that controls receptor function. Moreover, we characterize the conformational landscapes of the long third intracellular loop (ICL3), revealing its compact structural features and membrane-proximal localization in the basal state, where it acts as a negative allosteric regulator in transducer coupling. Furthermore, we identify multiful functional sub-states of ICL3 that are dynamically modulated by both kinase phosphorylation and drug efficacy. These findings offer previously underappreciated structural and dynamic insights into 2AAR function governed by both TMD and IDRs, and may open up new avenues for the development of better therapeutics.
]]></description>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Qiu, R.</dc:creator>
<dc:creator>Garces, A. M.</dc:creator>
<dc:creator>Hubner, H.</dc:creator>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Weikert, D.</dc:creator>
<dc:creator>Gmeiner, P.</dc:creator>
<dc:creator>Lerch, M. T.</dc:creator>
<dc:creator>Kobilka, B.</dc:creator>
<dc:date>2025-09-11</dc:date>
<dc:identifier>doi:10.1101/2025.09.10.675278</dc:identifier>
<dc:title><![CDATA[The role of intrinsically disordered domains in regulating G protein coupled receptor signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.11.675634v1?rss=1">
<title>
<![CDATA[
Expanding vaginal microbiome pangenomes via a custom MIDAS database reveals Lactobacillus crispatus accessory genes associated with cervical dysplasia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.11.675634v1?rss=1</link>
<description><![CDATA[
The vaginal microbiome plays a central role in reproductive health. Vaginal microbiome dysbiosis is associated with many adverse reproductive health outcomes, but most studies have focused on associations at the species level. The potential contribution of intraspecies microbial variation, especially gene content differences across bacterial strains, remains underexplored in reproductive health contexts. The Metagenomic Intra-Species Diversity Analysis (MIDAS) framework enables such analyses but depends on comprehensive reference databases. We constructed a MIDAS-compatible pangenome database from over 18,000 genomes in the Vaginal Microbiome Genome Collection (VMGC). Compared to the Genome Taxonomy Database (GTDB)-derived reference, the VMGC database expanded pangenomes of prevalent vaginal species, better capturing vaginal-specific intraspecies diversity. Applying this database to vaginal samples from a cervical dysplasia cohort, we identified thirteen Lactobacillus crispatus accessory genes significantly associated with cervical dysplasia, including a HicAB toxin-antitoxin system, three transcriptional regulators, and three phage-derived genes. These findings highlight the utility of body site-specific reference resources and shotgun metagenomic sequencing for uncovering intraspecies microbial variation relevant to reproductive health.

IMPORTANCEThe vaginal microbiome plays a critical role in reproductive health, and different bacteria from the same species can carry different genes that influence how the strains interact with the host and other microbes. These strain-level differences are often overlooked when microbiomes are analyzed only at the species level. Existing genomic reference databases are heavily biased toward gut and environmental bacteria, leaving the genetic diversity of vaginal microbes understudied. We built a specialized reference database from over 18,000 vaginal bacterial genomes that better reflects this diversity. We then applied this resource to quantify gene-level variation in vaginal samples from a cervical dysplasia cohort. Focusing on Lactobacillus crispatus, a prevalent and often beneficial vaginal species, we identified thirteen genes that were more common in women with cervical dysplasia than in controls. This work demonstrates that body site-specific genomic resources are essential for uncovering strain-level bacterial differences relevant to reproductive health.
]]></description>
<dc:creator>Dubin, C. A.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Oskotsky, T.</dc:creator>
<dc:creator>Golob, J. L.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2025-09-11</dc:date>
<dc:identifier>doi:10.1101/2025.09.11.675634</dc:identifier>
<dc:title><![CDATA[Expanding vaginal microbiome pangenomes via a custom MIDAS database reveals Lactobacillus crispatus accessory genes associated with cervical dysplasia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.10.675452v1?rss=1">
<title>
<![CDATA[
In situ Proteomics Unveils Specialized Domains for Extrasynaptic Signaling on Neuronal Cilia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.10.675452v1?rss=1</link>
<description><![CDATA[
Neuronal cilia have emerged as crucial signaling hubs, yet their molecular composition and integration with synaptic communication remain poorly understood. Using a newly developed Arl13b-TurboID mouse model, we achieved robust cilia-specific biotinylation and proteomic profiling across diverse tissues and cell types. Comparative proteomics revealed striking tissue-specific specialization, with neuronal cilia uniquely enriched in synaptic proteins, adhesion molecules, and neurotransmitter receptors. Surprisingly, several signaling and adhesion molecules localize to neuronal cilia in discrete nanodomains maintained by active retrieval mechanisms. In the mouse cortex, expansion microscopy revealed that the NMDA receptor subunit GluN1 is organized in nanodomains on neuronal ciliary membranes, which are precisely positioned to sample neurotransmitter efflux from neighboring glutamatergic synapses. These findings establish neuronal cilia as specialized extrasynaptic signaling platforms, with nanoscale organization enabling them to integrate local synaptic cues and modulate neuronal connectivity.
]]></description>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Trinh, V. N.</dc:creator>
<dc:creator>Lokesh, N. R.</dc:creator>
<dc:creator>Montecinos, C. K.</dc:creator>
<dc:creator>Pownall, M. E.</dc:creator>
<dc:creator>Kalocsay, M.</dc:creator>
<dc:creator>Nachury, M. V.</dc:creator>
<dc:date>2025-09-12</dc:date>
<dc:identifier>doi:10.1101/2025.09.10.675452</dc:identifier>
<dc:title><![CDATA[In situ Proteomics Unveils Specialized Domains for Extrasynaptic Signaling on Neuronal Cilia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.12.675939v1?rss=1">
<title>
<![CDATA[
H3N2 influenza virus tropism shifts to glycan receptors on tracheal ciliated cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.12.675939v1?rss=1</link>
<description><![CDATA[
Human H3N2 influenza viruses, introduced during the 1968 pandemic, have evolved to recognize human-type sialic acid-containing receptors (Neu5Ac2-6Gal) extended with at least three LacNAc (Gal{beta}1-4GlcNAc) repeats. To investigate this restriction in the context of virus attachment to the airway epithelium, we comprehensively analyzed the glycome of human nasal and tracheal epithelial cells. Using a synthetic N-glycan library that reflects the structural diversity of the human airway glycome, we found that only bi-antennary N-glycans with extended human-type receptors on at least one branch serve as receptors for the recent H3 hemagglutinins (HAs). Such receptors are found on tracheal epithelium but are deficient in nasal epithelium. Immunofluorescence analysis on human trachea reveals that recent H3 HAs preferentially attach to ciliated cells, consistent with single-cell RNA sequencing analysis indicating that these cells express glycosyltransferases that produce extended glycan chains. These findings suggest that H3N2 viruses have developed a tropism for tracheal ciliated cells. (146 words)
]]></description>
<dc:creator>Kikuchi, C.</dc:creator>
<dc:creator>Antonopoulos, A.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Biyasheva, A.</dc:creator>
<dc:creator>Chien, Y.-C.</dc:creator>
<dc:creator>Bharat, A.</dc:creator>
<dc:creator>McBride, R.</dc:creator>
<dc:creator>Nycholat, C. M.</dc:creator>
<dc:creator>Thompson, A. J.</dc:creator>
<dc:creator>Dell, A.</dc:creator>
<dc:creator>Khoo, K.-H.</dc:creator>
<dc:creator>Schleimer, R.</dc:creator>
<dc:creator>Haslam, S. M.</dc:creator>
<dc:creator>Paulson, J. C.</dc:creator>
<dc:date>2025-09-13</dc:date>
<dc:identifier>doi:10.1101/2025.09.12.675939</dc:identifier>
<dc:title><![CDATA[H3N2 influenza virus tropism shifts to glycan receptors on tracheal ciliated cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.09.675195v1?rss=1">
<title>
<![CDATA[
Evaluating the Contribution of Genome 3D Folding to Variation in Human Height Using Machine Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.09.675195v1?rss=1</link>
<description><![CDATA[
Genome-wide association studies (GWAS) have identified thousands of variants associated with complex traits, yet the majority lie in noncoding regions, making it difficult to determine their functional impact. Alterations to the three-dimensional (3D) spatial interactions among gene regulatory elements are increasingly recognized as a mechanism by which genetic variants influence gene expression. However, experimentally evaluating whether variants disrupt 3D-genome structure is not feasible at GWAS scale. To address this, we developed a computational framework that integrates GWAS summary statistics with predictions from the Akita sequence-based deep learning model of 3D chromatin contacts. We applied the framework to 9,917 genomic regions associated with human height, assessing both individual variants and haplotypes for their predicted impact on 3D genome architecture. Only a small fraction of height-associated haplotypes had substantial predicted disruption of 3D folding (17 regions, 0.17%, exceeded a disruption score of 0.1). Considering all common variants in a haplotype together generally produced greater perturbations than individual variants, but several highly divergent regions were driven by single variants. We highlight a variant that disrupts the binding motif at a confirmed CTCF binding site and is predicted to modify 3D genome contacts with the LCOR promoter, suggesting that 3D-genome-mediated disruption of gene regulation underlies the association with height. This work presents a scalable and interpretable strategy for integrating 3D genome modeling with GWAS, enabling investigation of this important regulatory mechanism in the connection of non-coding genetic variation to complex traits.
]]></description>
<dc:creator>Gu, W.</dc:creator>
<dc:creator>Gilbertson, E.</dc:creator>
<dc:creator>Baranzini, S. E.</dc:creator>
<dc:creator>Salem, R.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:date>2025-09-15</dc:date>
<dc:identifier>doi:10.1101/2025.09.09.675195</dc:identifier>
<dc:title><![CDATA[Evaluating the Contribution of Genome 3D Folding to Variation in Human Height Using Machine Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.10.675414v1?rss=1">
<title>
<![CDATA[
Anatomical integrity of the human cochlea estimated with optical coherence tomography for future clinical application 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.10.675414v1?rss=1</link>
<description><![CDATA[
The human cochlea is encased within the otic capsule, the densest bone in the body, posing significant challenges for anatomical imaging of cochlear structures. Because of difficult access and fragility of cochlear structures, our understanding of intracochlear anatomy has historically relied on postmortem histology. We thus have a limited understanding of human cochlear anatomy in its native, unfixed state. Clinical diagnostics for hearing loss, such as audiometry and otoacoustic emissions, offer functional assessments but fail to elucidate the often diverse underlying structural pathologies with any degree of precision. To address the critical need for assessing the human cochlear anatomy and associated pathologies without the risk of traumatizing cochlear structures, we imaged fresh cochleae in situ soon after death through the intact round window membrane with Optical Coherence Tomography (OCT) without inserting instruments inside or opening the cochlea. Micron-resolution OCT cross-sectional images of the human intracochlear structures were acquired and compared with corresponding histology systematically to aid in the identification of fine structural features and possible pathologies.

With OCT imaging, we observed varied anatomy of the organ of Corti, and developed a cochlear "integrity" rating system to differentiate healthy appearing cochleae from various pathological states. These results demonstrate the capability of OCT to non-traumatically visualize cochlear integrity, highlighting its potential as a diagnostic tool. This work shows promise in translating the ability to determine the likelihood of existing or lack of hair cells and supporting cells in live patients, which would enable appropriate targeted treatments.
]]></description>
<dc:creator>Secchia, P.</dc:creator>
<dc:creator>McHugh, C. I.</dc:creator>
<dc:creator>Cho, N. H.</dc:creator>
<dc:creator>O'Malley, J. T.</dc:creator>
<dc:creator>Zhu, M.</dc:creator>
<dc:creator>Dharmarajan, A.</dc:creator>
<dc:creator>Zosuls, A.</dc:creator>
<dc:creator>Feng, J. J.</dc:creator>
<dc:creator>Cheng, Y. S.</dc:creator>
<dc:creator>Puria, S.</dc:creator>
<dc:creator>Eckhard, A. H.</dc:creator>
<dc:creator>Nakajima, H. H.</dc:creator>
<dc:date>2025-09-16</dc:date>
<dc:identifier>doi:10.1101/2025.09.10.675414</dc:identifier>
<dc:title><![CDATA[Anatomical integrity of the human cochlea estimated with optical coherence tomography for future clinical application]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.12.675833v1?rss=1">
<title>
<![CDATA[
The Integrated Cellular and Molecular Landscape of Autoimmunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.12.675833v1?rss=1</link>
<description><![CDATA[
We performed a large-scale immunogenomic analysis of [~]13,000 transcriptomic profiles from 10 autoimmune diseases, integrating publicly available datasets from both blood and tissue. Using meta-analysis, we identified core immune mechanisms underlying autoimmunity, including strong interferon responses, inflammation, and adaptive immune suppression, alongside disease-specific signatures. To enhance biological interpretability, we derived higher-order immune features - such as cell type proportions, cytokine levels, pathway activity, transcription factor regulation, and miRNA activity - and organized them into 15 coherent immune modules. These modules enabled systematic cross-disease comparisons, revealing shared and distinct immunopathological patterns. The inflammation module, in particular, was associated with disease severity and predicted treatment response across multiple conditions. This modular framework offers a powerful tool for understanding immune dysregulation and advancing precision medicine in autoimmune diseases. To support reproducibility and enable others to build upon this work, we developed an interactive app to explore and download the complete dataset and associated results.
]]></description>
<dc:creator>Appierdo, R.</dc:creator>
<dc:creator>Pepe, G.</dc:creator>
<dc:creator>Helmer-Citterich, M.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Vallania, F.</dc:creator>
<dc:creator>Gherardini, P. F.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.12.675833</dc:identifier>
<dc:title><![CDATA[The Integrated Cellular and Molecular Landscape of Autoimmunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.16.676629v1?rss=1">
<title>
<![CDATA[
Virtual Reality and Tablet Cognitive Training Improve Attention and Academic Skills Without Dose Effects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.16.676629v1?rss=1</link>
<description><![CDATA[
We evaluated the cognitive and academic effects of a closed-loop video game delivered in virtual reality (VR) and tablet formats, at two different dosages, in a school-based setting. A total of 158 children aged 8-9 with a range of attention abilities completed 30 training sessions over 10 weeks. Compared to an expectancy-matched control group, both VR and tablet training led to significant improvements in teacher-rated inattention, performance-based attention tasks, and eye-tracking measures. While both VR and tablet versions of the intervention showed benefits on specific attention-related and academic outcomes, the VR version showed select advantages in both regards. Notably, intervention dose did not significantly moderate outcomes, suggesting that efficacy may depend more on reaching a threshold of engagement than on total duration. These findings demonstrate the utility and benefits of using each type of technology to enhance measures of cognitive and academic abilities as part of a regular school curriculum.
]]></description>
<dc:creator>Giannakopoulou, A.</dc:creator>
<dc:creator>Gordon, A. M.</dc:creator>
<dc:creator>Gallen, C. E.</dc:creator>
<dc:creator>Seaman, M.</dc:creator>
<dc:creator>Fedele, D.</dc:creator>
<dc:creator>Anguera, J. A.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.16.676629</dc:identifier>
<dc:title><![CDATA[Virtual Reality and Tablet Cognitive Training Improve Attention and Academic Skills Without Dose Effects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.18.677126v1?rss=1">
<title>
<![CDATA[
The eukaryotic replisome intrinsically generates asymmetric daughter chromatin fibers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.18.677126v1?rss=1</link>
<description><![CDATA[
DNA replication is molecularly asymmetric, due to distinct mechanisms for lagging and leading strand DNA synthesis. Whether chromatin assembly on newly replicated strands is also asymmetric remains unknown, as visualizing this short-lived state in cells is impossible. To circumvent this limitation, we combine in vitro reconstitution of the Saccharomyces cerevisiae DNA and chromatin replication machineries with replication-aware single-molecule chromatin footprinting, to study how chromatin is re-assembled on replicated DNA. Leveraging the non-destructive, single-molecule, and strand-specific nature of our data, we discover an intrinsic asymmetry in nucleosome positioning patterns and organization between lagging- and leading-strand chromatin created by the yeast replisome. This asymmetry is only partially restored upon addition of chromatin assembly factors involved in de novo histone deposition and the ATP-dependent chromatin remodeler Isw1a, implying that other regulatory factors must resolve this asymmetry in cells. In sum, our data reveal the complexity of chromatin re-establishment following DNA replication, and suggest an asymmetric chromatin assembly intermediate on each daughter chromatid. These pathways have implications for essential chromatin-templated processes such as DNA repair, transcription, and gene silencing at replication forks.
]]></description>
<dc:creator>Eckhardt, B. V.</dc:creator>
<dc:creator>Richter, H. J.</dc:creator>
<dc:creator>Rondeel, I.</dc:creator>
<dc:creator>Renduchintala, K.</dc:creator>
<dc:creator>Mattiroli, F.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:date>2025-09-18</dc:date>
<dc:identifier>doi:10.1101/2025.09.18.677126</dc:identifier>
<dc:title><![CDATA[The eukaryotic replisome intrinsically generates asymmetric daughter chromatin fibers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.18.676800v1?rss=1">
<title>
<![CDATA[
Metabolic and transcriptional adaptations to phagocytosis in microglia sustain their functionality and regenerative properties 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.18.676800v1?rss=1</link>
<description><![CDATA[
Phagocytosis of apoptotic cells, or efferocytosis, is a tightly regulated process that ensures tissue homeostasis and prevents mounting inflammatory responses. In the brain parenchyma, it is executed by microglia, which are encumbered by large numbers of apoptotic debris generated during development, in adult neurogenic niches, aging, and brain diseases. Emerging evidence suggest that phagocytosis is not limited to garbage disposal, but triggers adaptations in the phagocytes that may have a functional impact. To test it we developed an in vivo model of superphagocytosis induced by low cranial irradiation (LCI, 2Gy) that specifically induced apoptosis in the neurogenic niche of the adult hippocampus, synchronizing microglia in a phagocytic state within 6h and leading to full clearance by 24h. Single cell RNA sequencing and metabolomics revealed an unexpected oxidative stress in post-phagocytic microglia, accompanied by catabolic shutdown, mitochondrial remodeling, increased expression of galectin 3, and production of polyamines that led to cell death and compensatory proliferation. To test whether these changes impaired subsequent microglial phagocytosis, we used a glioblastoma model treated with sequential irradiation to induce tumor cell apoptosis. The phagocytosis efficiency of tumor-associated microglia/macrophages was comparable in the first and second apoptotic challenge, suggesting that the metabolic remodeling induced by phagocytosis was adaptive and destined to sustain their functionality. Finally, we assessed the functional impact of post-phagocytosis adaptations using galectin 3 deficient mice under LCI. We found that the recovery of the neurogenic niche after LCI strongly depended on galectin 3, demonstrating the regenerative capacity of post-phagocytic microglia. Overall, our data unveils the complexity of post-phagocytosis adaptations in microglia, underscoring their unexplored therapeutic potential in brain disorders.
]]></description>
<dc:creator>Marquez-Ropero, M.</dc:creator>
<dc:creator>Valero, J.</dc:creator>
<dc:creator>Cuesta-Puente, X.</dc:creator>
<dc:creator>Marquez-Galera, A.</dc:creator>
<dc:creator>Pereira-Iglesias, M.</dc:creator>
<dc:creator>Gonzalez-Dominguez, M.</dc:creator>
<dc:creator>Louail, A.</dc:creator>
<dc:creator>Beccari, S.</dc:creator>
<dc:creator>Lopez-Murillo, L.</dc:creator>
<dc:creator>San Juan, A.</dc:creator>
<dc:creator>Arrey, G.</dc:creator>
<dc:creator>Diaz-Aparicio, I.</dc:creator>
<dc:creator>Dubbelaar, M. L.</dc:creator>
<dc:creator>Ribera-Ramos, A.</dc:creator>
<dc:creator>Sarmiento Soto, M.</dc:creator>
<dc:creator>Ruiz, R.</dc:creator>
<dc:creator>Ayerra, L.</dc:creator>
<dc:creator>Vilas-Zornoza, A.</dc:creator>
<dc:creator>Cabrera, D.</dc:creator>
<dc:creator>van Liempd, S.</dc:creator>
<dc:creator>Garcia-Moreno, F.</dc:creator>
<dc:creator>Blomgren, K.</dc:creator>
<dc:creator>Gonzalez-Granero, S.</dc:creator>
<dc:creator>Garcia-Verdugo, J. M.</dc:creator>
<dc:creator>Falcon-Perez, J. M.</dc:creator>
<dc:creator>Aymerich, M. S.</dc:creator>
<dc:creator>Hambardzumyan, D.</dc:creator>
<dc:creator>Casafont, I.</dc:creator>
<dc:creator>Eggen, B. J. L.</dc:creator>
<dc:creator>Venero, J. L.</dc:creator>
<dc:creator>Lopez-Atalaya, J. P.</dc:creator>
<dc:creator>Sierra, A.</dc:creator>
<dc:date>2025-09-19</dc:date>
<dc:identifier>doi:10.1101/2025.09.18.676800</dc:identifier>
<dc:title><![CDATA[Metabolic and transcriptional adaptations to phagocytosis in microglia sustain their functionality and regenerative properties]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.16.676163v1?rss=1">
<title>
<![CDATA[
Single-sequence deep learning delivers crystal-quality models of covalent K-Ras G12 hotspot complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.16.676163v1?rss=1</link>
<description><![CDATA[
Structure-based design of covalent drugs has achieved tremendous success by understanding and leveraging the three-dimensional interactions between small-molecule drug candidates and their protein targets. However, this approach traditionally relies on high-resolution co-complex structures obtained by X-ray crystallography, NMR, or cryo-EM, methods that are time-consuming and resource-intensive. Here we show that Chai-1, a publicly available structure prediction tool that accepts user-defined ligands, is able to accurately predict covalent K-Ras(G12C) complexes without using a multiple sequence alignment (MSA). Chai-1 yields pocket-aligned RMSDs <2 [A] for chemically diverse K-Ras(G12C) inhibitors, ranging from ARS-853 to BBO-8520. In addition to the conventional acrylamide-based covalent K-Ras(G12C) inhibitors, Chai-1 with a covalent-bond restraint successfully reproduced the binding poses of covalent K-Ras(G12D) and K-Ras(G12S) inhibitors, while showing limitations in capturing chemical details such as accounting for leaving-groups, bond properties, and stereochemistry. Chai-1 also provides [~]40-fold higher throughput than state-of-the-art AlphaFold3 while maintaining comparable pose accuracy. Together, these findings establish Chai-1 as an accessible and computationally efficient tool for covalent protein-ligand co-complex structure prediction, with its covalent-restraint mode offering a unique solution to accelerate covalent drug discovery, especially for challenging targets beyond cysteine.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=79 SRC="FIGDIR/small/676163v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Jung, S.</dc:creator>
<dc:creator>Zheng, Q.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.16.676163</dc:identifier>
<dc:title><![CDATA[Single-sequence deep learning delivers crystal-quality models of covalent K-Ras G12 hotspot complexes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.19.677468v1?rss=1">
<title>
<![CDATA[
Deep Receptor Scanning Reveals General Sequence Constraints on GPCR Biosynthesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.19.677468v1?rss=1</link>
<description><![CDATA[
G protein-coupled receptors (GPCRs) mediate a variety of signaling pathways and are the most common pharmacological targets. While advances in structural biochemistry have provided deep functional insights into key receptors, many of the 800+ human GPCRs remain understudied. We introduce a versatile "deep receptor scanning" platform that can be used to experimentally characterize 766 human GPCRs and 174 known GPCR splice variants in parallel. We use this platform to quantitatively characterize the relative abundance of canonical and alternative receptor transcripts, their translational efficiency, and the plasma membrane expression of each receptor in the context of a recombinant pool of HEK293T cells expressing individual GPCRs. We then employ machine learning to identify specific structural features that modulate GPCR expression. This experimental platform and informatic approach are compatible with a variety of assays and can be used to efficiently explore the biochemical and pharmacological properties of the GPCRome.
]]></description>
<dc:creator>Tedman, A.</dc:creator>
<dc:creator>Goel, M.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Howard, M. K.</dc:creator>
<dc:creator>Chamness, L. M.</dc:creator>
<dc:creator>Bonifasi, A.</dc:creator>
<dc:creator>Adams, I.</dc:creator>
<dc:creator>Gallagher, J. M.</dc:creator>
<dc:creator>Penn, W. D.</dc:creator>
<dc:creator>Nemec, K.</dc:creator>
<dc:creator>McDonald, E. F.</dc:creator>
<dc:creator>Corman, B. N.</dc:creator>
<dc:creator>Robinson, J. P.</dc:creator>
<dc:creator>Post, C. B.</dc:creator>
<dc:creator>Clark, P. L.</dc:creator>
<dc:creator>Babu, M. M.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:creator>Kuntz, C. P.</dc:creator>
<dc:creator>Coyote-Maestas, W.</dc:creator>
<dc:creator>Schlebach, J. P.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.19.677468</dc:identifier>
<dc:title><![CDATA[Deep Receptor Scanning Reveals General Sequence Constraints on GPCR Biosynthesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.21.676939v1?rss=1">
<title>
<![CDATA[
An image-based CRISPR screen reveals splicing-mediated control of HP1α condensates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.21.676939v1?rss=1</link>
<description><![CDATA[
Heterochromatin Protein 1 (HP1) is a fundamental component of constitutive heterochromatin, forming subnuclear condensates whose regulation and function remain poorly understood. Here, we present an image-based CRISPR screen targeting nuclear factors that identifies splicing as a pivotal pathway regulating HP1 condensates. We discovered that unspliced intronic RNA modulates HP1 condensates by interacting co-transcriptionally with HP1. By modulating the intron content, RNA processing restricts HP1-RNA interactions at chromatin, thus enabling heterochromatin organization. Disruption of HP1 condensates due to enhanced interactions with unspliced RNA leads to loss of heterochromatin and the activation of stress response protective genes. We propose that RNA is a central component of heterochromatin that modulates HP1 condensates, and that RNA processing enzymes act as a surveillance mechanism for condensates by dynamically regulating the network of multi-valent interactions between RNA and chromatin factors. This model underscores the crosstalk between chromatin organization, transcription, and RNA processing, potentially governing broader nuclear functions.
]]></description>
<dc:creator>Wong, M. M.-K.</dc:creator>
<dc:creator>Zhou, S.</dc:creator>
<dc:creator>Carpenter, C.</dc:creator>
<dc:creator>Valbuena, R.</dc:creator>
<dc:creator>Priyadarshini, M.</dc:creator>
<dc:creator>Arya, A.</dc:creator>
<dc:creator>Rizvi, A.</dc:creator>
<dc:creator>Carswell-Crumpton, C.</dc:creator>
<dc:creator>Wileveau, A.</dc:creator>
<dc:creator>Lopez-Lopez, G.</dc:creator>
<dc:creator>Tycko, J.</dc:creator>
<dc:creator>Yao, D.</dc:creator>
<dc:creator>Spees, K.</dc:creator>
<dc:creator>Maynard, J.</dc:creator>
<dc:creator>Bassik, M. C.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Sanulli, S.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.21.676939</dc:identifier>
<dc:title><![CDATA[An image-based CRISPR screen reveals splicing-mediated control of HP1α condensates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.21.677611v1?rss=1">
<title>
<![CDATA[
Broad-Spectrum HIV-1 Detection and Neutralization via Multivalent Designer DNA Nanostructures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.21.677611v1?rss=1</link>
<description><![CDATA[
Early and accurate detection of HIV-1 remains a critical unmet need, particularly during the acute phase of infection when viral loads are low and transmission risk is highest. Here, we report a modular diagnostic and antiviral platform based on designer DNA nanostructures engineered for high-affinity recognition of the HIV-1 envelope glycoprotein (GP120). A custom DNA aptamer, termed HINT, was developed to bind GP120 across major HIV-1 subtypes (Groups M and P; subtypes A and B) with nanomolar affinity. To amplify binding strength, HINT aptamers were spatially patterned onto a net-shaped DNA nanostructure (DNA-NetHINT) that geometrically matches the trimeric GP120 spikes on the viral surface. Using multivalent interactions, the nanostructure enabled up to 104-fold improvement in binding affinity (sub-picomolar KD), confirmed by surface plasmon resonance. Integration of DNA-NetHINT into a paper-based lateral flow assay produced a low-cost, saliva-compatible self-testing device capable of detecting intact HIV-1 virions at concentrations as low as 328 viral copies per test, outperforming commercial fourth-generation rapid diagnostic tests. In addition to its diagnostic capabilities, the DNA-NetHINT construct exhibited potent antiviral activity, reducing pseudovirus infection with an EC50 of [~]1.8 nM, nearly 1,000-fold more effective than free aptamers. This work demonstrates a dual-function DNA nanotechnology platform that enables both ultrasensitive HIV-1 detection and entry inhibition. The approach is broadly applicable to other enveloped viruses and represents a promising step toward next-generation molecular theranostics for infectious disease management.
]]></description>
<dc:creator>Umrao, S.</dc:creator>
<dc:creator>Dwivedy, A.</dc:creator>
<dc:creator>Zheng, M.</dc:creator>
<dc:creator>Anirudhan, V.</dc:creator>
<dc:creator>Parlatan, U.</dc:creator>
<dc:creator>Henrich, T.</dc:creator>
<dc:creator>Cunningham, B.</dc:creator>
<dc:creator>Demirci, U.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.21.677611</dc:identifier>
<dc:title><![CDATA[Broad-Spectrum HIV-1 Detection and Neutralization via Multivalent Designer DNA Nanostructures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.21.677627v1?rss=1">
<title>
<![CDATA[
Bicistronic CAR T-cells Against CD70 & Active Integrin β2 Overcome Antigen Heterogeneity and Preserve Safety in Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.21.677627v1?rss=1</link>
<description><![CDATA[
The surface antigen landscape of acute myeloid leukemia (AML) displays significant heterogeneity and overlap with healthy hematopoietic cells. This imparts a substantial hurdle to the development of AML-targeting chimeric antigen receptor (CAR) T-cells that can avoid on- target, off-tumor toxicity. Here, we develop a dual-antigen targeting CAR-T against CD70 and the active conformation of integrin {beta}2 (aITGB2), each previously reported as promising AML targets due to minimal off-tumor expression. We show an OR-gated approach for these antigens significantly increases the proportion of AML blasts that can be targeted, in part using a novel ex vivo co-culture method to restore surface protein homeostasis following a freeze-thaw cycle. We test dual-targeting CAR-T constructs with different combinations of costimulatory domains, identifying constructs with superior anti-tumor cytotoxicity in vitro against AML cell line and patient-derived xenograft models. We further show significantly improved in vivo tumor clearance and survival for a dual-targeting CAR in murine models of AML tumor heterogeneity. Finally, we show that this dual-targeting CAR does not increase off-tumor toxicity, especially against hematopoietic stem and progenitor cells. Together, these findings demonstrate a promising clinically-translatable approach for the treatment of AML without the notable toxicity liabilities associated with other leading CAR-T targets for this disease.
]]></description>
<dc:creator>Kang, A. S.</dc:creator>
<dc:creator>Johnson, H.</dc:creator>
<dc:creator>Lei, N.</dc:creator>
<dc:creator>Wong, J.</dc:creator>
<dc:creator>Razi, N.</dc:creator>
<dc:creator>Izgutdina, A.</dc:creator>
<dc:creator>Kasap, C.</dc:creator>
<dc:creator>Chilakapati, N.</dc:creator>
<dc:creator>Rivera, J.</dc:creator>
<dc:creator>Phojanakong, P.</dc:creator>
<dc:creator>Serrano, J. A. C.</dc:creator>
<dc:creator>Salangsang, F.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Logan, A. C.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Huang, B. J.</dc:creator>
<dc:creator>Wiita, A. P.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.21.677627</dc:identifier>
<dc:title><![CDATA[Bicistronic CAR T-cells Against CD70 & Active Integrin β2 Overcome Antigen Heterogeneity and Preserve Safety in Acute Myeloid Leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.22.676989v1?rss=1">
<title>
<![CDATA[
Defective proviruses cause T cell reprogramming through promoter exaptation in HIV-1 infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.22.676989v1?rss=1</link>
<description><![CDATA[
People living with HIV (PLWH) on antiretroviral therapy (ART) accumulate primarily defective proviral sequences in genomes of often clonally expanded CD4+ HIV-1 target cells. The majority of viral-derived DNA is transcriptionally active and preferentially found at distinct genomic loci suggesting a selective process driven by integration site-specific crosstalk between viral and host sequences. Focusing on one of the most prominent selected integration loci, the BTB Domain and CNC Homolog 2 (BACH2) gene, we here show mechanistic insights how CD4+ T cells are functionally reprogrammed via exaptation of provirus-derived regulatory sequences during long-term ART. Using a cellular model of BACH2-integrated proviruses, we find that proviral transcription drives aberrant BACH2 protein levels that escape autoregulatory feedback and impose BACH2-dependent transcriptomic changes. By mimicking these changes in primary CD4+ T lymphocytes, we observe that BACH2 drives reprogramming of cells toward a proliferative, precursor memory-like type. These reprogrammed CD4+ T cells possess traits of immune evasion and cellular survival that are signatures of persistent HIV reservoir cells in PLWH. Inhibition of provirus transcriptional activity can mitigate exaptation, suggesting a strategy to offset HIV-driven differentiation and expansion of CD4+ T cells. Finally, our data suggest that provirus exaptation at a second prominently selected proviral integration gene, the Signal Transducer And Activator of Transcription 5B (STAT5B) gene, drives a contrary, effector-like T cell fate, suggesting a multifaceted impact of exaptation on immune homeostasis. Overall, our data suggest that transcriptionally active proviruses, even if structurally defective, modulate target cells through insertional activation of integration genes, a process which we postulate to contribute to the complex immune modulation and dysregulation experienced by ART-suppressed PLWH.
]]></description>
<dc:creator>Hamann, M. V.</dc:creator>
<dc:creator>Brauckmann, L.</dc:creator>
<dc:creator>Schwarz, C.</dc:creator>
<dc:creator>Spohn, M.</dc:creator>
<dc:creator>Stoeck, R.</dc:creator>
<dc:creator>Leddy, S.</dc:creator>
<dc:creator>Frouard, J.</dc:creator>
<dc:creator>Adiba, M.</dc:creator>
<dc:creator>Thiele, F.</dc:creator>
<dc:creator>Virdi, S.</dc:creator>
<dc:creator>Grundhoff, A.</dc:creator>
<dc:creator>Feschotte, C.</dc:creator>
<dc:creator>Roan, N. R.</dc:creator>
<dc:creator>Tolosa, E.</dc:creator>
<dc:creator>Lange, U. C.</dc:creator>
<dc:date>2025-09-22</dc:date>
<dc:identifier>doi:10.1101/2025.09.22.676989</dc:identifier>
<dc:title><![CDATA[Defective proviruses cause T cell reprogramming through promoter exaptation in HIV-1 infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.22.677864v1?rss=1">
<title>
<![CDATA[
Neuroanatomical Correlates of Negative Symptoms in Schizophrenia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.22.677864v1?rss=1</link>
<description><![CDATA[
BackgroundSchizophrenia is characterized by widespread structural brain abnormalities, but associations between structural abnormalities and negative symptom severity are not well understood. Negative symptoms have been conceptualized in a hierarchical structure of two second-order dimensions--motivation and pleasure (MAP) and expression (EXP)--and five first-order domains: anhedonia, avolition, and asociality (MAP), and blunted affect and alogia (EXP). A better understanding of the neural circuitry underlying negative symptom dimensions and domains is important given their reported association with poor functional outcome and lack of available treatments.

Study DesignThe meta-analysis included 1,591 individuals with schizophrenia across 16 samples with structural imaging and Scale for Assessment of Negative Symptoms data. The study generated correlations of cortical thickness and subcortical volumes with the negative symptom dimensions and domains.

Study resultsNegative symptoms showed mainly negative associations with cortical thickness and subcortical volumes. The effect sizes were small but there was a pattern of associations in predominantly frontal lobe cortical thickness and limbic subcortical volumes. The regional correlation patterns of cortical thickness and subcortical volumes with symptom domains support the conceptualized hierarchical structure of negative symptoms: correlations of MAP domains were stronger with the MAP than EXP dimension, and vice versa. Exploratory analyses with receptor densities further supported the hierarchy.

ConclusionOur findings reveal small but consistent associations between negative symptom dimensions and predominantly prefrontal region cortical thickness, and limbic region volumes.

These findings advance our understanding of the network of anatomical regions that may contribute to the severity of negative symptoms in schizophrenia.
]]></description>
<dc:creator>Kamalakannan, S. M. V.</dc:creator>
<dc:creator>Male, A. G.</dc:creator>
<dc:creator>Yilanli, M.</dc:creator>
<dc:creator>Lella, A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Quide, Y.</dc:creator>
<dc:creator>Green, M. J.</dc:creator>
<dc:creator>Cairns, M. J.</dc:creator>
<dc:creator>Carr, V. J.</dc:creator>
<dc:creator>Catts, S.</dc:creator>
<dc:creator>Henskens, F. A.</dc:creator>
<dc:creator>Jablensky, A.</dc:creator>
<dc:creator>Loughland, C.</dc:creator>
<dc:creator>Michie, P.</dc:creator>
<dc:creator>Mowry, B.</dc:creator>
<dc:creator>Pantelis, C.</dc:creator>
<dc:creator>Shall, U.</dc:creator>
<dc:creator>Scott, R. J.</dc:creator>
<dc:creator>Belger, A.</dc:creator>
<dc:creator>Bustillo, J.</dc:creator>
<dc:creator>Lim, K.</dc:creator>
<dc:creator>Ford, J. M.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>Preda, A.</dc:creator>
<dc:creator>Mueller, B.</dc:creator>
<dc:creator>Potkin, S. G.</dc:creator>
<dc:creator>Satterhwaite, T. D.</dc:creator>
<dc:creator>Gur, R. C.</dc:creator>
<dc:creator>Gur, R. E.</dc:creator>
<dc:creator>Banaj, N.</dc:creator>
<dc:creator>Vecchio, D.</dc:creator>
<dc:creator>Piras, F.</dc:creator>
<dc:creator>Piras, F.</dc:creator>
<dc:creator>Ehrlich, S.</dc:creator>
<dc:creator>Bernardoni, F.</dc:creator>
<dc:creator>Borgwardt, S.</dc:creator>
<dc:creator>Cobia, D.</dc:creator>
<dc:creator>Alpert, K.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Agartz, I.</dc:creator>
<dc:creator>Jonsson, E. G.</dc:creator>
<dc:creator>Kaiser, S.</dc:creator>
<dc:creator>Clotet, E. P.</dc:creator>
<dc:creator>Salvador, R.</dc:creator>
<dc:creator>Jaramillo, C. L.</dc:creator>
<dc:date>2025-09-23</dc:date>
<dc:identifier>doi:10.1101/2025.09.22.677864</dc:identifier>
<dc:title><![CDATA[Neuroanatomical Correlates of Negative Symptoms in Schizophrenia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.19.677274v1?rss=1">
<title>
<![CDATA[
RNAcentral in 2026: Genes and literature integration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.19.677274v1?rss=1</link>
<description><![CDATA[
RNAcentral was founded in 2014 to serve as a comprehensive database of non-coding RNA sequences. It began by providing a single unified interface to more specialised resources, and now contains 45 million sequences. It has grown beyond providing a single interface to many specialised resources and now provides several services and analyses. These include secondary structure prediction with R2DT, sequence search, and analysis with Rfam. Since its last publication in 2021, RNAcentral has developed two major features. First, literature integration with the development of LitScan and LitSumm. LitScan automatically identifies and links relevant publications to RNA entries, while LitSumm uses natural language processing to generate functional summaries from the literature. Together, these tools address the critical challenge of connecting sequence data with scattered functional knowledge across thousands of publications. Secondly, RNAcentral has created gene level entries. Gene level entries represent a large structural change to RNAcentral. While RNAcentral previously organized data exclusively at the sequence level, we now group related transcripts into gene-centric views. This allows researchers to explore all isoforms, splice variants, and related sequences for a gene in a unified interface, better reflecting biological organization and facilitating comparative analyses. RNAcentral is freely available at: https://rnacentral.org.
]]></description>
<dc:creator>Green, A. F.</dc:creator>
<dc:creator>Ribas, C. E.</dc:creator>
<dc:creator>Jandalala, I.</dc:creator>
<dc:creator>Muston, P.</dc:creator>
<dc:creator>O'Cathail, C.</dc:creator>
<dc:creator>Cochrane, G.</dc:creator>
<dc:creator>Ernst, C.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:creator>Madrigal, P.</dc:creator>
<dc:creator>Attrill, H.</dc:creator>
<dc:creator>Marygold, S.</dc:creator>
<dc:creator>Lancet, D.</dc:creator>
<dc:creator>Dobzinski, N.</dc:creator>
<dc:creator>Chan, P. P.</dc:creator>
<dc:creator>Lowe, T. M.</dc:creator>
<dc:creator>Bruford, E. A.</dc:creator>
<dc:creator>Seal, R. L.</dc:creator>
<dc:creator>Hermjakob, H.</dc:creator>
<dc:creator>Panneerselvam, K.</dc:creator>
<dc:creator>Finn, R. D.</dc:creator>
<dc:creator>Gurbich, T. A.</dc:creator>
<dc:creator>Griffiths-Jones, S.</dc:creator>
<dc:creator>Fromm, B.</dc:creator>
<dc:creator>Peterson, K. J.</dc:creator>
<dc:creator>Sordyl, D.</dc:creator>
<dc:creator>Bujnicki, J. M.</dc:creator>
<dc:creator>Velankar, S.</dc:creator>
<dc:creator>Appasamy, S. D.</dc:creator>
<dc:creator>Ganguly, S.</dc:creator>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>He, S.</dc:creator>
<dc:creator>Rutherford, K. M.</dc:creator>
<dc:creator>Wood, V.</dc:creator>
<dc:creator>Lovering, R. C.</dc:creator>
<dc:creator>Picardi, E.</dc:creator>
<dc:creator>Ontiveros, N.</dc:creator>
<dc:creator>Huang, L.</dc:creator>
<dc:creator>Miao, Z.</dc:creator>
<dc:creator>Petrov, A. S.</dc:creator>
<dc:creator>McCann, H.</dc:creator>
<dc:creator>Cavalleri, E.</dc:creator>
<dc:creator>Mesiti, M.</dc:creator>
<dc:creator>Rivas, E.</dc:creator>
<dc:creator>Szikszai</dc:creator>
<dc:date>2025-09-24</dc:date>
<dc:identifier>doi:10.1101/2025.09.19.677274</dc:identifier>
<dc:title><![CDATA[RNAcentral in 2026: Genes and literature integration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.23.678127v1?rss=1">
<title>
<![CDATA[
In vivo genome-wide CRISPR screens in human T cells to enhance T cell therapy for solid tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.23.678127v1?rss=1</link>
<description><![CDATA[
Large-scale CRISPR screening in human T cells holds significant promise for identifying genetic modifications that can enhance cellular immunotherapy. However, many genetic regulators of T cell performance in solid tumors may not be readily revealed in vitro. In vivo screening in tumor-bearing mice offers greater physiological relevance, but has historically been limited by low intratumoral T cell recovery. Here, we developed a new model system that achieves significantly higher human T cell recovery from tumors, enabling genome-wide in vivo screens with small numbers of mice. Tumor-infiltrating T cells in this model exhibit hallmarks of dysfunction compared to matched splenic T cells, creating an ideal context for screening for genetic modifiers of T cell activity in the tumor microenvironment. Using this platform, we performed two genome-wide CRISPR knockout screens to identify genes regulating T cell intratumoral abundance and effector function (e.g., IFN-{gamma} production). The intratumoral abundance screen uncovered the P2RY8-G13 GPCR signaling pathway as a negative regulator of human T cell infiltration into tumors. The effector function screen identified GNAS (Gs), a central signaling mediator downstream of multiple GPCRs that sense different suppressive ligands, as a key regulator of T cell dysfunction in tumors. Targeted GNAS knockout rendered T cells resistant to multiple suppressive cues and significantly improved therapeutic performance across diverse solid tumor models. Moreover, combinatorial knockout of P2RY8 (trafficking) and GNAS (effector function) further enhanced overall tumor control, demonstrating that genetic modifications targeting distinct T cell phenotypes can be combined to improve therapeutic potency. This flexible and scalable in vivo screening platform can be adapted to diverse tumor models and pooled CRISPR libraries, enabling future discovery of genetic strategies that equip T cell therapies to overcome barriers imposed by solid tumors.
]]></description>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Chen, P. A.</dc:creator>
<dc:creator>Urs, E.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Arce, M.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Seo, J.</dc:creator>
<dc:creator>Kale, N.</dc:creator>
<dc:creator>LaFlam, T. N.</dc:creator>
<dc:creator>Peng, F.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Allen, G.</dc:creator>
<dc:creator>Eyquem, J. N.</dc:creator>
<dc:creator>Fuh, K.</dc:creator>
<dc:creator>Dodgson, S.</dc:creator>
<dc:creator>Cyster, J. G.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Carnevale, J.</dc:creator>
<dc:date>2025-09-24</dc:date>
<dc:identifier>doi:10.1101/2025.09.23.678127</dc:identifier>
<dc:title><![CDATA[In vivo genome-wide CRISPR screens in human T cells to enhance T cell therapy for solid tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.23.678137v1?rss=1">
<title>
<![CDATA[
Dissecting Gene Regulatory Networks Governing Human Cortical Cell Fate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.23.678137v1?rss=1</link>
<description><![CDATA[
Human cortical neurogenesis involves conserved and specialized developmental processes during a restricted window of prenatal development. Radial glia (RG) neural stem cells shape cortical cell diversity by giving rise to excitatory neurons, oligodendrocytes, and astrocytes, as well as olfactory bulb interneurons (INs) and a recently characterized population of cortical INs1,2. Complex genetic programs orchestrated by transcription factor (TF) circuits govern the balance between self-renewal and differentiation, and between different cell fates3-8. Despite progress in measuring gene regulatory network activity during human cortical development9-12, functional studies are required to evaluate the roles of TFs and effector genes in human RG lineage progression. Here we establish a human primary culture system that allows sensitive discrimination of cell fate dynamics and apply single cell clustered regularly interspaced short palindromic repeats interference (CRISPRi) screening13,14 to examine the transcriptional and cell fate consequences of 44 TFs active during cortical neurogenesis. We identified multiple TFs, with novel roles in cortical neurogenesis, including ZNF219, previously uncharacterized, that represses neural differentiation and NR2E1 and ARX that have opposing roles in regulating RG lineage plasticity and progression across developmental stages. We also uncovered convergent effector genes downstream of multiple TFs enriched in neurodevelopmental and neuropsychiatric disorders and observed conserved mechanisms of RG lineage plasticity across primates. We further uncovered a postmitotic role for ARX in safeguarding IN subtype specification through repressing LMO1. Our study provides a framework for dissecting regulatory networks driving cell fate consequences during human neurogenesis.
]]></description>
<dc:creator>Ding, J. W.</dc:creator>
<dc:creator>Kim, C. N.</dc:creator>
<dc:creator>Ostrowski, M. S.</dc:creator>
<dc:creator>Abeykoon, Y.</dc:creator>
<dc:creator>Pavlovic, B. J.</dc:creator>
<dc:creator>Wallace, J. L.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:date>2025-09-24</dc:date>
<dc:identifier>doi:10.1101/2025.09.23.678137</dc:identifier>
<dc:title><![CDATA[Dissecting Gene Regulatory Networks Governing Human Cortical Cell Fate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.23.678138v1?rss=1">
<title>
<![CDATA[
Comparing Neanderthal introgression maps reveals core agreement but substantial heterogeneity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.23.678138v1?rss=1</link>
<description><![CDATA[
Statistical methods to identify Neanderthal ancestry in modern human genomes rest on varying assumptions and inputs. Nonetheless, most studies of introgression use only a single method to define Neanderthal ancestry. Due to a lack of "ground truth," we have a limited understanding of the accuracy, comparative strengths and weaknesses, and the sensitivity of downstream conclusions for these methods. Here, we performed large-scale comparisons of genome-wide introgression maps from 12 representative Neanderthal introgression detection algorithms. These span methods that consider archaic and human reference genomes not from Africa (ArchaicSeeker2, CRF, DICAL-ADMIX), only archaic genomes (S*, Sprime, HMM, SARGE, ARGWeaver-D), only human reference genomes, including from Africa (IBDmix), or simulated data (ArchIE). Our results highlight a core set of regions predicted by nearly all methods, as well as substantial heterogeneity in commonly used Neanderthal introgression maps. Furthermore, we find that downstream analyses may result in different conclusions depending on the map used. Thus, we recommend careful consideration of map(s) chosen for an analysis and support the use of multiple maps to ensure robustness of conclusions. We make integrated prediction sets available, enabling further understanding of Neanderthal introgressions legacy on modern humans.
]]></description>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Velazquez-Arcelay, K.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:date>2025-09-24</dc:date>
<dc:identifier>doi:10.1101/2025.09.23.678138</dc:identifier>
<dc:title><![CDATA[Comparing Neanderthal introgression maps reveals core agreement but substantial heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.22.677949v1?rss=1">
<title>
<![CDATA[
Identity-stable multi-animal tracking using bidirectional segmentation with object-level memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.22.677949v1?rss=1</link>
<description><![CDATA[
Tracking animal behavior in naturalistic settings is essential for understanding social dynamics and their neural underpinnings. Pose estimation methods can produce accurate keypoints using framewise inference. However, post hoc tracking steps often struggle to maintain consistent identity over time, particularly during close and rapid social interactions between visually similar animals. We present a pipeline for bidirectional video object segmentation (VOS) to correct identity swaps with much less manual annotation effort, addressing the prohibitive cost of identity correction of pose estimation data in long recordings and large cohorts. Our approach makes use of a state-of-the-art VOS algorithm, Cutie, which leverages both pixel- and object-level representations across multiple memory timescales. By comparing segmentation masks from independent forward and reverse inference runs, we identify localized zones of disagreement and flag them for manual review. When applied to more than 160 hours of dyadic vole interaction videos, our method reduces identity swaps by two orders of magnitude compared to typical pose estimation workflows and requires review of less than 0.3% of frames per video to achieve identity error-free segmentation masks and aligned keypoints. Our approach generalizes to social interactions involving three or more animals, with scalability constrained primarily by behavioral complexity (e.g., complete occlusion of multiple individuals). Our method enables scalable, long-term tracking of unmarked animals in group settings and provides a practical foundation for more naturalistic studies of social behavior. To lower the barrier for researchers facing similar tracking challenges, we provide an accessible graphical user interface for general use.
]]></description>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Quine, K.</dc:creator>
<dc:creator>Jordan, A.</dc:creator>
<dc:creator>Dasari, S.</dc:creator>
<dc:creator>Manoli, D. S.</dc:creator>
<dc:creator>Kirst, C.</dc:creator>
<dc:date>2025-09-25</dc:date>
<dc:identifier>doi:10.1101/2025.09.22.677949</dc:identifier>
<dc:title><![CDATA[Identity-stable multi-animal tracking using bidirectional segmentation with object-level memory]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.23.678178v1?rss=1">
<title>
<![CDATA[
Getting a head start: Craniofacial heterochrony in marsupials involves dynamic changes to molecular and cellular mechanisms underlying neural crest development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.23.678178v1?rss=1</link>
<description><![CDATA[
The neural crest is a vertebrate innovation central to craniofacial development and evolution. While the gene regulatory networks guiding neural crest development are well characterized, the mechanisms generating species-specific craniofacial diversity remain poorly understood. Marsupials provide a unique model for studying neural crest plasticity, having evolved accelerated patterns of craniofacial development during embryogenesis. This adaptation arises in response to marsupials being born altricial after a short gestation yet require well-developed mouthparts to attach to a teat and continue development in the pouch. However, how marsupials achieve this heterochronic shift in neural crest development is largely unknown. In this study, we investigate the cellular and molecular mechanisms underlying their distinct heterochrony, revealing that marsupials produce dense pre-migratory aggregates of neural crest cells which undergo collective migration as epithelial-like sheets, potentially facilitating rapid establishment of the facial prominences. These cellular behaviours are unique amongst amniotes but resemble patterns in anamniotes which similarly exhibit accelerated craniofacial development to support early feeding. Marsupials appear to have evolved a similar mechanism of neural crest migration to facilitate their developmental heterochrony. These findings suggest that vertebrate neural crest migration may be shaped by the pace of craniofacial development during embryogenesis rather than phylogeny, providing new perspectives on neural crest plasticity and the developmental mechanisms driving craniofacial diversity across vertebrates.
]]></description>
<dc:creator>Newton, A. H.</dc:creator>
<dc:creator>Farley, E. R.</dc:creator>
<dc:creator>Major, A. T.</dc:creator>
<dc:creator>Hutchison, J. C.</dc:creator>
<dc:creator>Lawrence, B. M.</dc:creator>
<dc:creator>Sears, K. E.</dc:creator>
<dc:creator>Renfree, M. B.</dc:creator>
<dc:creator>Couzens, A. M. C.</dc:creator>
<dc:creator>Shaw, G.</dc:creator>
<dc:creator>Ord, S.</dc:creator>
<dc:creator>Schneider, R. A.</dc:creator>
<dc:creator>Pask, A. J.</dc:creator>
<dc:date>2025-09-25</dc:date>
<dc:identifier>doi:10.1101/2025.09.23.678178</dc:identifier>
<dc:title><![CDATA[Getting a head start: Craniofacial heterochrony in marsupials involves dynamic changes to molecular and cellular mechanisms underlying neural crest development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.25.678472v1?rss=1">
<title>
<![CDATA[
Childhood brain tumours instruct cranial haematopoiesis and immunotolerance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.25.678472v1?rss=1</link>
<description><![CDATA[
Recent research has revealed a remarkable role for immunosurveillance in healthy and diseased brains, dispelling the notion that this organ is a passive immune-privileged site1-3. Better understanding of how this immunosurveillance operates could improve the treatment of neurological diseases. Here, using a novel genetically engineered mouse model of ZFTA-RELA ependymoma4-a childhood brain tumour-we characterised an immune circuit between the tumour and antigen presenting, haematopoietic stem/progenitor cells (HSPCs) in the skull bone marrow. The presentation of antigens in the cerebrospinal fluid (CSF) by HSPCs to CD4+ T cells, biased HSPC lineages toward myelopoiesis and polarised CD4+ T-cells to regulatory T cells (T- regs), culminating in tumour immunotolerance. Remarkably, a single infusion of antibodies directed against cytokines enriched in the CSF of mice bearing ZFTA-RELA ependymomas, choroid plexus carcinomas or Group-3 medulloblastoma-all aggressive childhood brain tumours-disrupted this process and caused profound tumour regression. These data unmask a mechanism by which skull bone marrow-derived HSPCs and CD4+ T cells cooperate to promote the immunotolerance of childhood brain tumours. Antibodies that disrupt this immunosurveillance could prove an effective therapy for these cancers that are less toxic than current treatments.
]]></description>
<dc:creator>Cooper, E. A.</dc:creator>
<dc:creator>Posner, D. A.</dc:creator>
<dc:creator>Lee, C. Y. C.</dc:creator>
<dc:creator>Hu, L.</dc:creator>
<dc:creator>Bonner, S.</dc:creator>
<dc:creator>Taylor, J. T.</dc:creator>
<dc:creator>Baldwin, O.</dc:creator>
<dc:creator>Jimenez-Guerrero, R.</dc:creator>
<dc:creator>Masih, K. E.</dc:creator>
<dc:creator>Wickham Rahrmann, K.</dc:creator>
<dc:creator>Eigenbrood, J.</dc:creator>
<dc:creator>Ngo, G.</dc:creator>
<dc:creator>Franklin, V. N. R.</dc:creator>
<dc:creator>D'Santos, C.</dc:creator>
<dc:creator>Mair, R.</dc:creator>
<dc:creator>Santarius, T.</dc:creator>
<dc:creator>Craven, C.</dc:creator>
<dc:creator>Jalloh, I.</dc:creator>
<dc:creator>Moreno-Vicente, J.</dc:creator>
<dc:creator>Halim, T.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:creator>Swartling, F. J.</dc:creator>
<dc:creator>Khan, J.</dc:creator>
<dc:creator>Clatworthy, M. R.</dc:creator>
<dc:creator>Gilbertson, R. J.</dc:creator>
<dc:date>2025-09-26</dc:date>
<dc:identifier>doi:10.1101/2025.09.25.678472</dc:identifier>
<dc:title><![CDATA[Childhood brain tumours instruct cranial haematopoiesis and immunotolerance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.25.678463v1?rss=1">
<title>
<![CDATA[
Joint Mapping of Chromatin Accessibility and Targeted Proteomics in HER2-expressing Breast Cancer Systems 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.25.678463v1?rss=1</link>
<description><![CDATA[
HER2 proteoforms promote therapeutic resistance and aggressiveness in HER2-positive breast cancer, yet their epigenetic consequences remain poorly defined. Here, we establish EpiBlot, a joint assay incorporating a customized plateATAC-seq workflow that minimizes sample inputs with single-cell western blotting to concurrently profile chromatin accessibility with protein and proteoform expression. We applied our method to engineered MCF7 cells expressing HER2 proteoforms - full-length p185HER2 or truncated 611-CTF -, where we evaluated the impact of such proteoforms on the epigenetic and protein profiles after lapatinib or doxorubicin exposure. Expression of 611-CTF elicits pervasive chromatin remodeling, whereas p185HER2 provokes only modest accessibility shifts under the same treatments. EpiBlot reveals that treatment with doxorubicin drives extensive genome-wide accessibility changes, while lapatinib treatment produces limited global effects but unmasks proteoform-specific responses. Concordance between chromatin accessibility and protein abundance is moderate, underscoring complex regulatory coupling. Extending this dual-modality approach to HER2-low patient-derived organoids uncovers distinct chromatin states and reveals a subpopulation of triple-negative breast-cancer cells expressing truncated HER2 proteoforms. We anticipate that EpiBlot will highlight the value of multimodal profiling with proteoform identification for dissecting tumor heterogeneity and therapeutic response in cancer.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=198 SRC="FIGDIR/small/678463v2_ufig1.gif" ALT="Figure 1">
View larger version (45K):
org.highwire.dtl.DTLVardef@673263org.highwire.dtl.DTLVardef@11d28c5org.highwire.dtl.DTLVardef@15fa336org.highwire.dtl.DTLVardef@1d4426b_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Fomitcheva Khartchenko, A.</dc:creator>
<dc:creator>Lam, T.</dc:creator>
<dc:creator>Goldhammer, N.</dc:creator>
<dc:creator>Bont, M.</dc:creator>
<dc:creator>Warhadpande, S.</dc:creator>
<dc:creator>Rosenbluth, J.</dc:creator>
<dc:creator>Herr, A.</dc:creator>
<dc:date>2025-09-27</dc:date>
<dc:identifier>doi:10.1101/2025.09.25.678463</dc:identifier>
<dc:title><![CDATA[Joint Mapping of Chromatin Accessibility and Targeted Proteomics in HER2-expressing Breast Cancer Systems]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.25.678618v1?rss=1">
<title>
<![CDATA[
High resolution spatial profiling of the hematopoietic landscape of the murine lung 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.25.678618v1?rss=1</link>
<description><![CDATA[
Our current understanding of blood cell development and functionality stems primarily from the investigation of adult bone marrow (BM) and the fetal liver prenatally. However, emerging evidence highlights the lung as a previously underappreciated residence for hematopoietic cells. While a diversity of cells specific to the BM are known to promote the maturation and trafficking of hematopoietic cells, how the lung niche influences the development and functionality of resident cells is not known. Spatial in situ transcriptomics enables accurate mapping of cell identities and interactions within intact tissue, providing insights not accessible by dissociated single-cell profiling. Here, we present a high-resolution spatial transcriptomic atlas of the healthy adult murine lung placing specific emphasis on the hemato-endothelial landscape of this organ. As a case study, we developed a semi-automatic workflow to explicitly identify and curate rare - often multinucleated - megakaryocytes, requiring a combination of hex-binning spatial enrichment of canonical markers, expert curation, and cell boundary merging to correct for segmentation artifacts. We then characterized the spatial neighborhoods of megakaryocytes, illustrating their topological embedding within vascular, stromal, and immune microenvironments. Finally, we demonstrated the utility of this dataset for hypothesis-driven signaling studies by examining ligand-receptor interactions across pathways including BMP, VEGF, and ECM-integrin signaling. Together, this work defines the lung-blood niche and advances our understanding of the organ-specific properties of blood cells. We also provide a high-resolution spatial reference for the murine lung and demonstrate how targeted spatial in situ transcriptomics enable focused case studies of rare hematopoietic niches.

KEY POINTSO_LIThis work represents the highest resolution gene expression mapping of the spatial symbiosis between the hematopoietic and pulmonary systems.
C_LIO_LIPulmonary megakaryocytes localize within distinct vascular and stromal neighborhoods.
C_LI
]]></description>
<dc:creator>Skvir, N. J.</dc:creator>
<dc:creator>Ysasi, A. B.</dc:creator>
<dc:creator>Jarzabek, V.</dc:creator>
<dc:creator>Yeung, A. K.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Dries, R. J.</dc:creator>
<dc:creator>Murphy, G. J.</dc:creator>
<dc:date>2025-09-27</dc:date>
<dc:identifier>doi:10.1101/2025.09.25.678618</dc:identifier>
<dc:title><![CDATA[High resolution spatial profiling of the hematopoietic landscape of the murine lung]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.25.678166v1?rss=1">
<title>
<![CDATA[
Daple-FLT3 (CCDC88C-FLT3) gene fusion requires the coiled-coil domain for maximal activation and pericentrosomal localization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.25.678166v1?rss=1</link>
<description><![CDATA[
Gene fusions are stable protein products often occurring from chromosomal rearrangements. These chimeric proteins typically contain distinct molecular entities from each parent gene, and thus, create a product with altered or aberrant function. Gene fusions are frequently found in cancers, including Leukemia. Here, we characterize the kinase activity and subcellular distribution of the Daple-FLT3 (CCDC88C-FLT3) fusion oncoprotein--a rare, but recurrent gene fusion found in patients with hematological malignancies. The protein contains the FLT3 kinase domain and is activated without ligand stimulation. This leads to activation in STAT5a, AKT, and MAPK signaling, which can be modulated by the tyrosine kinase inhibitors (TKIs) sorafenib, quizartinib, and to a lesser degree, imatinib. Moreover, fusion of this kinase domain to Daple facilitates its localization to the pericentrosomal space and enhances kinase activation. These findings provide evidence that targeting Daple-FLT3 outside of its kinase domain may be a complementary approach with TKI therapy.

Key PointsO_LIDaple-FLT3 fusion proteins contain a constitutively active kinase domain, activating distinct signaling molecules in cells
C_LIO_LICoiled-coil domain on Daple is dispensable for kinase activation, but necessary for maximal activation
C_LI
]]></description>
<dc:creator>Kao, D.</dc:creator>
<dc:creator>Acquazzino, M.</dc:creator>
<dc:creator>Ibarra, A.</dc:creator>
<dc:creator>Valenzuela, E.</dc:creator>
<dc:creator>Mai, H.</dc:creator>
<dc:creator>Aguilar, K.</dc:creator>
<dc:creator>Stieglitz, E.</dc:creator>
<dc:creator>Ear, J.</dc:creator>
<dc:date>2025-09-29</dc:date>
<dc:identifier>doi:10.1101/2025.09.25.678166</dc:identifier>
<dc:title><![CDATA[Daple-FLT3 (CCDC88C-FLT3) gene fusion requires the coiled-coil domain for maximal activation and pericentrosomal localization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.25.678627v1?rss=1">
<title>
<![CDATA[
Perinatal Shifts in Fecal-Derived Metabolites and Associations with Postpartum Depression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.25.678627v1?rss=1</link>
<description><![CDATA[
IntroductionResearch on maternal depression is largely limited to static, blood-derived biomarkers in the postpartum period and mechanistic targets derived from populations outside the physiological contexts of pregnancy and postpartum, resulting in critical gaps in understanding context-specific mechanisms underlying this debilitating condition.

ObjectivesTo examine temporal shifts in the maternal gut metabolome and associations between pregnancy-specific shifts and postpartum depression (PPD).

MethodsWe applied untargeted metabolomics (UPLC-MS/MS) to fecal samples collected from participants (N= 25) enrolled in the Maternal and Infant NutriTion (MINT) study. Random forest analysis was used to identify key pathways and metabolites contributing to temporal shifts. Fold change analysis and paired t-tests were used to quantify the magnitude and significance of metabolite changes.

Associations between pregnancy-specific changes and PPD (Edinburgh Postnatal Depression Scale at 6 weeks) were identified using Pearsons correlation.

ResultsLipid, amino acid, and xenobiotic metabolism emerged as core pathways driving temporal changes in the maternal gut metabolome. The most pronounced shifts occurred from 35 weeks gestation to postpartum, with 55 metabolites significantly altered compared to 24 from 24-to 35 weeks gestation and 26 from 24 weeks gestation to postpartum. Of the 29 metabolites associated with PPD; 68.9% were metabolic intermediates, primarily involved in lipid and amino acid metabolism (58.6%).

ConclusionsThis study provides some of the first evidence of temporal shifts in the maternal gut metabolome and associations with PPD, highlighting the importance of lipid and amino acid metabolism and laying the groundwork for future multi-omics research.
]]></description>
<dc:creator>Longoria, K. D.</dc:creator>
<dc:creator>Cassidy, M. L.</dc:creator>
<dc:creator>Lewis, K. A.</dc:creator>
<dc:creator>Garcia, S. R.</dc:creator>
<dc:creator>Franco-Rocha, O. Y.</dc:creator>
<dc:creator>Dube, S.</dc:creator>
<dc:creator>Nguyen, T. C.</dc:creator>
<dc:creator>Flowers, E.</dc:creator>
<dc:creator>Widen, E. M.</dc:creator>
<dc:date>2025-09-29</dc:date>
<dc:identifier>doi:10.1101/2025.09.25.678627</dc:identifier>
<dc:title><![CDATA[Perinatal Shifts in Fecal-Derived Metabolites and Associations with Postpartum Depression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.25.678692v1?rss=1">
<title>
<![CDATA[
Hepatic cytochrome P450 endoplasmic reticulum-associated degradation (ERAD): Topological determinants and cellular partnerships that dictate the preferential P450 proteolytic sorting into macroautophagy rather than UPS. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.25.678692v1?rss=1</link>
<description><![CDATA[
Many N-terminally endoplasmic reticulum (ER)-anchored cytochrome P450 proteins (P450s) turn over proteolytically via ER-associated degradation (ERAD), others via ER-to-lysosomal-associated degradation (ERLAD), and yet others via both pathways. What precisely dictates their differential proteolytic turnover is unknown. Herein, we employed rabbit liver CYPs 1A1 and 1A2, which reportedly reside in liquid-disorded (ld)- and detergent-resistant, liquid-ordered (lo)-ER-microdomains, respectively, governed by their specific N-terminal (NT) signal-anchor (SA) subdomains. We now report that this precise SA-dependent ER-topology not only determines the proclivity of CYP1A1 towards ERAD and CYP1A2 towards ERLAD, but also their differential lifespans. We further document that the detergent-resistant lo-ER-membranes (DRMs) are morphologically quite similar to mitochondria-associated ER-membranes (MAMs), documented cellular platforms for autophagic-initiation complexes. DRMs and MAMs, composed of saturated fatty-acids, glycosphingolipids and cholesterol, harbor many common morphological markers including the ER-specific prohibitin, erlin-1. Herein employing SURF, a split fluorogenic bifunctional complementation assay, we show that intracellularly, erlin-1 and CYP1A2 interact closely via their ER NT-SAs. siRNA-knockdown (KD) of erlin-1 in HepG2-cells, not only relocated CYP1A2 from DRMs to non-DRMs, but also impaired its ERLAD, resulting in insoluble cellular CYP1A2 aggregates. Upon erlin-1 KD, CYP1A2 ERLAD could be rescued by co-expression of a siRNA-resistant intact erlin-1 or just its NT-1-30 residue SA-domain. Our findings are the first to reveal that the CYP1A2 lifespan and preferential proclivity towards ERLAD is determined by its close association with erlin-1 within DRMs/MAMs. As proof of concept, we document that the ERLAD-proclivity of CYP2B1 is also similarly dependent upon erlin-1-DRM-association.

SIGNIFICANCE STATEMENTThe endoplasmic reticulum (ER)-anchored cytochromes P450 (P450s) incur ER-associated degradation (ERAD) and/or ER-to-lysosomal-associated degradation (ERLAD). What determines their preferential proteolytic turnover is unknown. Here, employing P450s CYP1A1 and CYP1A2 that reside in their N-terminal signal anchor-determined liquid-disordered (ld)- and detergent-resistant, liquid-ordered (lo)-ER-microdomains, respectively, we documented that in HepG2-cells these ER-microdomains determine the CYP1A proteolytic preferences for ERAD vs ERLAD as well as their lifespans. More importantly, we discovered that its intimate interaction with the ER-specific prohibitin erlin-1 colocalized in these lo-ER-microdomains, specifically dictates CYP1A2s ERLAD-preference. Accordingly, siRNA-elicited erlin-1-knockdown disrupted CYP1A2-ERLAD, which was rescued upon coexpression of either a siRNA-resistant erlin-1 or just its N-terminal 1-30 residues. As proof of concept, we document similar characteristics for CYP2B1, another ERLAD-targeted P450.
]]></description>
<dc:creator>Hong, X.</dc:creator>
<dc:creator>He, L.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Shu, X.</dc:creator>
<dc:creator>Correia, M. A.</dc:creator>
<dc:date>2025-09-29</dc:date>
<dc:identifier>doi:10.1101/2025.09.25.678692</dc:identifier>
<dc:title><![CDATA[Hepatic cytochrome P450 endoplasmic reticulum-associated degradation (ERAD): Topological determinants and cellular partnerships that dictate the preferential P450 proteolytic sorting into macroautophagy rather than UPS.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.26.678822v1?rss=1">
<title>
<![CDATA[
The Integrated Stress Response Pathway Improves Aged Murine Muscle Stem Cell Activation and in vivo Regeneration. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.26.678822v1?rss=1</link>
<description><![CDATA[
For efficient regeneration, muscle stem cells (MuSCs) transition out of quiescence through a series of progressively more activated states. During MuSC aging, transition through the earliest steps is the slowest and delayed, with the molecular regulators that govern this transition not well characterized. By analyzing the dynamic changes of MuSCs at the molecular (scRNA-Seq and Cell Painting) and phenotypic (heteromotility) level at single cell resolution we found that the Integrated Stress Response (ISR) Pathway is a critical regulator of MuSC transition states. Aged MuSCs have increased baseline ISR activity in quiescence that does not increase during activation to levels observed in adult MuSCs. Rapid and transient pharmacological ISR activation in vitro was sufficient to increase aged MuSC activation rate and migratory behavior as well as alter the transcriptional states toward a younger phenotype. ISR activation also improved aged MuSC potency and aged mouse muscle regeneration in vivo. Therefore, pharmacological activation of the ISR has therapeutic potential to improve MuSC function and skeletal muscle repair during aging.
]]></description>
<dc:creator>Brown, A. D.</dc:creator>
<dc:creator>Scaramozza, A.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Mahin, S.</dc:creator>
<dc:creator>Suresh, N.</dc:creator>
<dc:creator>Tsusaka, T.</dc:creator>
<dc:creator>Eliazer, S.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Feeley, B.</dc:creator>
<dc:creator>Brack, A. S.</dc:creator>
<dc:date>2025-09-29</dc:date>
<dc:identifier>doi:10.1101/2025.09.26.678822</dc:identifier>
<dc:title><![CDATA[The Integrated Stress Response Pathway Improves Aged Murine Muscle Stem Cell Activation and in vivo Regeneration.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.29.679324v1?rss=1">
<title>
<![CDATA[
Engineering Inducible Cell Fate Transitions by Harnessing Epigenetic Silencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.29.679324v1?rss=1</link>
<description><![CDATA[
During development, cell-cell communication induces a series of cell fate transitions that are maintained by epigenetic gene regulation. Here, we harness endogenous epigenetic silencing machinery to develop synthetic circuits that induce stable gene expression changes. Using synthetic Notch receptors that control the chromatin regulators KRAB and Dnmt3L, we developed input-controlled switches capable of inducing self-sustaining silencing of target loci. We used these modules to construct circuits in which combinatorial inputs specifically direct a choice among multiple alternative cell fates. These epigenetic silencing switches can also be inverted to yield input-induced sustained activation of a target gene. We demonstrate that this epigenetic memory switch can be used to drive morphological fate changes, in response to transient cell signals, that remain stable over many cell divisions, as is observed in development. These synthetic epigenetic circuits represent an important step towards engineering cell populations capable of coordinated multi-cell fate decisions.
]]></description>
<dc:creator>Campos, O. A.</dc:creator>
<dc:creator>Migliara, A.</dc:creator>
<dc:creator>Toda, S.</dc:creator>
<dc:creator>Lopez, P.</dc:creator>
<dc:creator>Lim, W. A.</dc:creator>
<dc:creator>Almeida, R.</dc:creator>
<dc:date>2025-09-30</dc:date>
<dc:identifier>doi:10.1101/2025.09.29.679324</dc:identifier>
<dc:title><![CDATA[Engineering Inducible Cell Fate Transitions by Harnessing Epigenetic Silencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.29.679346v1?rss=1">
<title>
<![CDATA[
Accelerated Tempo of Cortical Neurogenesis in Down Syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.29.679346v1?rss=1</link>
<description><![CDATA[
Down syndrome (DS), caused by trisomy 21 (TS21), is the most common genetic cause of intellectual disability1,2. The neurological impacts of DS first manifest during prenatal development through reduced radial glia (RG) neural stem cell proliferation, reduced cortical volume and imbalanced cortical cell types3-6. However, the developmental mechanisms underlying altered cortical neurogenesis in DS remain elusive. Here we show by high-throughput lineage tracing in organotypic culture that TS21 accelerates RG lineage progression, driving premature production of cortical inhibitory neurons (INs) and oligodendrocytes. Somatic lineage coupling connects dysregulated neurogenic tempo to altered cellular composition in the adult DS brain. Finally, lineage-resolved differential expression reveals elevated interferon responses specifically in RG biased to producing INs. Together, our findings link TS21 genomic abnormalities to candidate molecular pathways and developmental mechanisms altering the cellular landscape in DS with therapeutic relevance.
]]></description>
<dc:creator>Ding, J. W.</dc:creator>
<dc:creator>Kim, C. N.</dc:creator>
<dc:creator>Steyert, M. R.</dc:creator>
<dc:creator>Yuan, A. T.</dc:creator>
<dc:creator>Shin, D.</dc:creator>
<dc:creator>Ivanov, D.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:date>2025-09-30</dc:date>
<dc:identifier>doi:10.1101/2025.09.29.679346</dc:identifier>
<dc:title><![CDATA[Accelerated Tempo of Cortical Neurogenesis in Down Syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.29.677401v1?rss=1">
<title>
<![CDATA[
Clinical cure of chronic hepatitis B is dependent on activation and perpetuation of robust CD4+ T cell responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.29.677401v1?rss=1</link>
<description><![CDATA[
Chronic infection with hepatitis B virus (HBV), a major global pathogen, often leads to immune-mediated progressive liver injury and liver cancer. While seroclearance of the surface antigen (HBsAg) defines clinical cure and reduces disease-associated risks, HBsAg clearance is rarely observed and remains therapeutically elusive. Here we overcome some of the challenges to studying immune mechanisms of HBsAg clearance in chronic hepatitis B (CHB) using our mouse model of age-dependent HBsAg clearance and persistence, and samples from our BeNEG-DO clinical trial that provided longitudinal PBMCs from patients who either cleared HBsAg or retained stable HBsAg levels after stopping nucleos(t)ide analog therapy. We show that young mice fail to clear HBsAg and have impaired ability to efficiently initiate and sustain HBV-specific CD4+ T cell responses. We also demonstrate a role for CD4+ T cells in hepatic leukocyte organization and cytotoxicity, and in HBV-specific CD8+ T cell cytotoxicity and HBsAg clearance. Upstream of the CD4+ T cell response, we reveal that hepatic dendritic cells, particularly cDC2s, direct effective CD4+ T cell activation and differentiation. Studies in CHB patients identified immune features of HBsAg clearance that overlap with the mouse model, including TH1 and cytotoxic CD4+ T cell activation and CD8+ T cell cytotoxic effector function. These findings identify an essential role for potent CD4+ T cell activation in the clinical cure of CHB and illuminate potential immunotherapeutic targets for enhancing CD4+ T cell responses to achieve greater HBsAg clearance rates.

One Sentence SummaryUsing a mouse model of hepatitis B and longitudinal PBMCs from patients with chronic hepatitis B, we identify shared mechanisms of HBsAg seroclearance.
]]></description>
<dc:creator>Jespersen, J. M.</dc:creator>
<dc:creator>Avanesyan, L.</dc:creator>
<dc:creator>Publicover, J.</dc:creator>
<dc:creator>Carey, N. D.</dc:creator>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Edwards, A. W.</dc:creator>
<dc:creator>Stenske, S.</dc:creator>
<dc:creator>Shin, J.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Simone, M.</dc:creator>
<dc:creator>Chew, N. W.</dc:creator>
<dc:creator>Wong, N.</dc:creator>
<dc:creator>Trilok, S.</dc:creator>
<dc:creator>Erickson, A.</dc:creator>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Loo, C. P.</dc:creator>
<dc:creator>Stec, M.</dc:creator>
<dc:creator>Anderson, M.</dc:creator>
<dc:creator>Cloherty, G.</dc:creator>
<dc:creator>Sarkar, M.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Wakil, A. E.</dc:creator>
<dc:creator>Segal, M. R.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Cooper, S.</dc:creator>
<dc:creator>Baron, J. L.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.09.29.677401</dc:identifier>
<dc:title><![CDATA[Clinical cure of chronic hepatitis B is dependent on activation and perpetuation of robust CD4+ T cell responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.29.679218v1?rss=1">
<title>
<![CDATA[
A mechanical origin for implantation defects in embryos from aged females 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.29.679218v1?rss=1</link>
<description><![CDATA[
Women over 35 experience a marked reduction in fertility. The origin of these fertility defects appears to reside in the implantation capacity of the embryo itself, but the mechanistic basis of this impairment is not well-understood. Here, we identify a core mechanical defect in embryos from aged mothers that impairs the process of implantation. Using mouse models, we find that reproductive aging yields increased contractility in the extra-embryonic trophectoderm, the outer epithelial tissue responsible for mediating uterine attachment and embryo implantation. This hypercontractile state elevates tissue surface tension and viscosity in the blastocyst, culminating in defective spreading during implantation. Enhanced contractility is necessary and sufficient for this age-related defect in implantation, and early embryo mechanics can be used to predict successful implantation for embryos from both young and aged mothers. Our work represents a potential foundation for improving embryo selection in Assisted Reproductive Technologies to resolve age-related defects in female fertility.
]]></description>
<dc:creator>Cavanaugh, K. E.</dc:creator>
<dc:creator>Franco-Onate, M.</dc:creator>
<dc:creator>Laird, D. J.</dc:creator>
<dc:creator>Oakes, P. W.</dc:creator>
<dc:creator>Alert, R.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.09.29.679218</dc:identifier>
<dc:title><![CDATA[A mechanical origin for implantation defects in embryos from aged females]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.29.679252v1?rss=1">
<title>
<![CDATA[
Single-cell-level digital twins for preterm birth prevention strategies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.29.679252v1?rss=1</link>
<description><![CDATA[
Digital twin models can accelerate therapeutic development by enabling low-risk testing of candidate interventions. In preterm labor (PTL), a major pregnancy complication where clinical trials face unique ethical and financial barriers, digital twins are especially valuable for evaluating new therapies targeting immune dysfunctions driving PTL. Yet, current models lack single-cell resolution, limiting detection of cell-type-specific mechanisms, off-target effects, and the design of personalized interventions. We present Simulated Immunome Modeling of Clinical Outcomes (SIMCO), a single-cell-level digital twin framework that models immunomodulatory treatment effects on the timing of labor using immunome-wide, single-cell simulations. SIMCOs digital twins are trained and validated on a newly generated mass cytometry atlas of the pregnant immunome exposed to nine candidate drugs preselected for PTL prevention. Applying SIMCO to an independent cohort of pregnant individuals, we simulate treatment effects on gestational length, screening for candidate drugs that delay labor timing and providing system-level mechanistic insight for each drug candidate. Tetrahydrofolate, maprotiline, and the combination of aspirin and lansoprazole emerged as top candidates for PTL prevention, delaying labor onset primarily through enhanced mTOR signaling in innate immune cells and attenuated JAK/STAT signaling in naive CD4 T cells. The codebase is available at https://github.com/ofondeur/SIMCO/.
]]></description>
<dc:creator>Einhaus, J.</dc:creator>
<dc:creator>Neidlinger, P.</dc:creator>
<dc:creator>Fondeur, O.</dc:creator>
<dc:creator>Sato, M.</dc:creator>
<dc:creator>Anronikov, A.</dc:creator>
<dc:creator>Miyazaki, K.</dc:creator>
<dc:creator>Amar, J. N.</dc:creator>
<dc:creator>Ando, K.</dc:creator>
<dc:creator>Badea, V.</dc:creator>
<dc:creator>Gaudilliere, D. K.</dc:creator>
<dc:creator>Sabayev, M.</dc:creator>
<dc:creator>Feyaerts, D.</dc:creator>
<dc:creator>Diop, M.</dc:creator>
<dc:creator>Tsai, A. S.</dc:creator>
<dc:creator>Cambriel, A.</dc:creator>
<dc:creator>Ganio, E. A.</dc:creator>
<dc:creator>Lagarde, R.</dc:creator>
<dc:creator>O'Kelly, E.</dc:creator>
<dc:creator>Stelzer, I. A.</dc:creator>
<dc:creator>Hedou, J.</dc:creator>
<dc:creator>Wong, R. J.</dc:creator>
<dc:creator>Blumenfeld, Y. J.</dc:creator>
<dc:creator>Lyell, D. J.</dc:creator>
<dc:creator>Shaw, G. M.</dc:creator>
<dc:creator>Oskotsky, T. T.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Giudice, L. C.</dc:creator>
<dc:creator>Stevenson, D. K.</dc:creator>
<dc:creator>Aghaeepour, N.</dc:creator>
<dc:creator>Gaudilliere, B.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.09.29.679252</dc:identifier>
<dc:title><![CDATA[Single-cell-level digital twins for preterm birth prevention strategies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.01.679551v1?rss=1">
<title>
<![CDATA[
Mechanism of phosphoinositide regulation of lysosomal pH via inhibition of CLC-7 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.01.679551v1?rss=1</link>
<description><![CDATA[
AbstractLysosomes process cellular waste and coordinate responses to metabolic challenge. Central to lysosomal homeostasis are phosphoinositide lipids, key signaling molecules which establish organelle identity, regulate membrane dynamics and are tightly linked to the pathophysiology and therapy of lysosomal storage disorders, neurodegeneration, and cancer. Phosphatidylinositol 3,5- bisphosphate (PI(3,5)P2) interacts with multiple lysosomal membrane proteins and plays a critical role in regulating lysosomal pH by directly inhibiting the chloride/proton antiporter ClC-7, though the molecular mechanism of this inhibition remains unclear. Here, using a combination of functional, structural, and computational analysis, we demonstrate that PI(3,5)P2 binding dramatically remodels the structure of ClC-7 by inducing close association between cytosolic and transmembrane domains. Disease-causing mutations show increased transport activity through loss of PI(3,5)P2 binding and subsequent inhibition. Conversely, ClC-7 activation is correlated with dissociation and increased disorder of the cytoplasmic domain along with novel transmembrane domain conformations, revealing a mechanistic link between specific lysosomal lipids, transporter regulation, and the enigmatic basis of the ClC-7 slow gate.
]]></description>
<dc:creator>Hilton, J. K.</dc:creator>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Sefah, E.</dc:creator>
<dc:creator>Deme, J. C.</dc:creator>
<dc:creator>Parker, J. L.</dc:creator>
<dc:creator>Langton, M. J.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>Lea, S. M.</dc:creator>
<dc:creator>Newstead, S.</dc:creator>
<dc:creator>Mindell, J. A.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679551</dc:identifier>
<dc:title><![CDATA[Mechanism of phosphoinositide regulation of lysosomal pH via inhibition of CLC-7]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.01.679841v1?rss=1">
<title>
<![CDATA[
NOVA: a novel R-package enabling multi-parameter analysis and visualization of neural activity in MEA recordings 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.01.679841v1?rss=1</link>
<description><![CDATA[
Multielectrode array (MEA) technology enables simultaneous recording of electrical signals from neuronal networks, producing complex datasets. Current analytical approaches typically examine a limited number of metrics such as mean firing rate and synchronicity, leaving much of the data underutilized. To address this gap, we created NOVA (Neural Output Visualization and Analysis), an accessible R-based computational tool for comprehensive MEA data interpretation and visualization. NOVA integrates dimensionality reduction through principal component analysis, hierarchical clustering with heatmap generation, and temporal trajectory mapping of network activity patterns. Our code offers both a userfriendly pipeline requiring minimal coding background as well as customizable advanced plotting modules for experienced users. Validation experiments using primary cortical neurons during development and pharmacological manipulation demonstrated NOVAs capacity to detect subtle activity shifts overlooked by conventional methods. Notably, our unbiased approach identified network burst duration as a stronger contributor to activity variance than commonly reported firing rate metrics, exemplifying NOVAs utility for discovering meaningful patterns and generating data-driven hypotheses.
]]></description>
<dc:creator>Escoubas, C. C.</dc:creator>
<dc:creator>Guney, E.</dc:creator>
<dc:creator>Miravet, A. T.</dc:creator>
<dc:creator>Magee, N.</dc:creator>
<dc:creator>Phua, R.</dc:creator>
<dc:creator>Ruggero, D.</dc:creator>
<dc:creator>Molofsky, A. V.</dc:creator>
<dc:creator>Weiss, W. A.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679841</dc:identifier>
<dc:title><![CDATA[NOVA: a novel R-package enabling multi-parameter analysis and visualization of neural activity in MEA recordings]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.01.679860v1?rss=1">
<title>
<![CDATA[
Few-Shot Classification of Cryo-EM Micrographs Using Triplet Loss Embeddings 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.01.679860v1?rss=1</link>
<description><![CDATA[
Micrograph quality assessment in cryo-electron microscopy (cryo-EM) presents a significant challenge: users must either manually screen thousands of micrographs or expend substantial computational resources processing potentially low-quality data. While few-shot learning has been applied to particle picking and subtomogram classification in cryo-EM, its application to micrograph-level quality assessment remains largely unexplored. We present a framework combining few-shot learning with cryo-EM micrograph classification using triplet loss embeddings. By combining real-space and Fourier-space information in an embedding network trained with triplet loss, we achieve competitive performance across multiple EMPIAR datasets using as few as 1-5 labeled examples per class. Our approach demonstrates improvements over traditional cross-entropy training, which often collapses to predicting only the majority class in the few-shot regime. These results suggest a practical framework for rapid adaptation of automated micrograph screening to new experimental conditions with minimal manual labeling.
]]></description>
<dc:creator>Ho, A.</dc:creator>
<dc:creator>Sung, B.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Tsai, F. T. F.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679860</dc:identifier>
<dc:title><![CDATA[Few-Shot Classification of Cryo-EM Micrographs Using Triplet Loss Embeddings]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.01.679895v1?rss=1">
<title>
<![CDATA[
Cerebellar outputs for rapid directional refinement of forelimb movement 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.01.679895v1?rss=1</link>
<description><![CDATA[
Much of our interaction with the world relies on the ability to move our limbs with speed and precision. The cerebellum is critical for movement coordination, yet how outputs from the cerebellum continually guide the limb and whether discrete pathways differentially contribute to adjusting motor output remain unclear. Using intersectional viral approaches in mice, we identify two spatially intermingled yet anatomically distinct cerebellar populations that drive the forelimb either toward or away from the body. Neural recordings reveal cerebellar activity that correlates with and precedes these opposing directional changes in limb movement. Both cerebellar output pathways influence motor neuron and muscle activity within milliseconds, producing reliable effects on limb trajectory despite substantial underlying variability in muscle recruitment patterns. Our findings disentangle a subtype organization to cerebellar limb control, revealing a subcortical circuit basis for online directional refinement during movement execution.
]]></description>
<dc:creator>Thanawalla, A. R.</dc:creator>
<dc:creator>Wilcox, O.</dc:creator>
<dc:creator>Rhee, E.</dc:creator>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Huang, K. W.</dc:creator>
<dc:creator>Yusufi, R.</dc:creator>
<dc:creator>Saklaway, D.</dc:creator>
<dc:creator>Nagamori, A.</dc:creator>
<dc:creator>Conner, J. M.</dc:creator>
<dc:creator>Chen, A. I.</dc:creator>
<dc:creator>Azim, E.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679895</dc:identifier>
<dc:title><![CDATA[Cerebellar outputs for rapid directional refinement of forelimb movement]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.02.680065v1?rss=1">
<title>
<![CDATA[
Directed evolution of a stem-helix targeting antibody enables MERS-CoV cross-neutralization through enhanced binding affinity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.02.680065v1?rss=1</link>
<description><![CDATA[
Broadly neutralizing antibodies (bnAbs) targeting conserved regions of the betacoronavirus spike are important for pan-betacoronavirus protection and pandemic preparedness. Here, we report on the isolation of a human monoclonal antibody, CC65.1, from a SARS-CoV-2 convalescent donor that targets the conserved S2 stem helix region. CC65.1 neutralizes various sarbecoviruses, including SARS-CoV-2, and binds the MERS-CoV spike but lacks MERS-CoV-neutralizing activity due to insufficient binding affinity. We utilized directed evolution to enhance the binding affinity of CC65.1 for the MERS-CoV S2 stem helix, yielding engineered antibody variants with newly acquired MERS-CoV-neutralizing activity. High-resolution structural analysis reveals key paratope mutations that optimize binding and stabilize epitope engagement. Our findings demonstrate the potential of rational antibody engineering to expand bnAb breadth across divergent betacoronaviruses. This work supports the development of engineered bnAbs and S2-targeted vaccines for broad betacoronavirus countermeasures and highlights strategies to achieve cross-lineage immunity for future pandemic threats.

Author SummaryThe persistent emergence of new SARS-CoV-2 variants of concern that evade neutralizing antibody responses and other zoonotic betacoronaviruses with pandemic potential have provided strong motivation to develop broadly neutralizing antibodies (bnAbs) that target more conserved regions of the spike protein in sarbecoviruses and other betacoronaviruses. Here, we employed a directed evolution strategy to engineer the sarbecovirus-neutralizing antibody CC65.1, which targets the conserved S2 stem helix, to enhance its binding affinity with the MERS-CoV stem helix region, thereby conferring MERS-CoV neutralization. High-resolution structural studies of engineered CC65.1 revealed that key mutations reshape the paratope to better accommodate and stabilize the MERS-CoV S2 stem helix, resulting in increased binding affinity and neutralization potency. This study emphasizes the critical role of affinity maturation in expanding neutralization breadth and provides valuable insights for design of bnAbs to prevent and treat pandemic threats by betacoronaviruses.
]]></description>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Yuan, M.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Limbo, O.</dc:creator>
<dc:creator>Song, G.</dc:creator>
<dc:creator>Zhao, F.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>He, W.-t.</dc:creator>
<dc:creator>Capozzola, T.</dc:creator>
<dc:creator>Callaghan, S.</dc:creator>
<dc:creator>Avillion, G.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Beutler, N.</dc:creator>
<dc:creator>Yong, P.</dc:creator>
<dc:creator>Anzanello, F.</dc:creator>
<dc:creator>Rogers, T. F.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Jardine, J. G.</dc:creator>
<dc:creator>Wilson, I. A.</dc:creator>
<dc:creator>Andrabi, R.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.02.680065</dc:identifier>
<dc:title><![CDATA[Directed evolution of a stem-helix targeting antibody enables MERS-CoV cross-neutralization through enhanced binding affinity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.02.679816v1?rss=1">
<title>
<![CDATA[
Poising and connectivity of emergent human developmental enhancers in the transition from naive to primed pluripotency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.02.679816v1?rss=1</link>
<description><![CDATA[
In primed human pluripotent stem cells (hPSCs) resembling post-implantation epiblast, numerous lineage-specific enhancers assume the poised chromatin state, co-marked by H3K4me1 and Polycomb-associated H3K27me3 histone modifications. In contrast, poised enhancers (PEs) are scarce in naive hPSCs that model pre-implantation epiblast. PEs form abundant chromosomal contacts with developmental genes, but when these contacts emerge, how their formation relates to enhancer poising and their functional significance remains incompletely understood. Here, we devise high-resolution, PE-targeted Capture Hi-C to generate a comprehensive atlas of PE chromosomal contacts in the time course of hPSC transition from the naive to primed state. We find that enhancer poising emerges early in the transition, while the contacts show diverse dynamics that is only partially coupled to poising. PROTAC-induced degradation of Polycomb Repressive Complex 2 (PRC2) early in the transition weakens PE connectivity, while inhibition of its H3K27 methyltransferase activity does not, suggesting a non-catalytic role of Polycomb in supporting PE contacts. Notably, PE contacts persist after developmental activation or ectopic CRISPRa targeting and can mediate long-range gene induction. Together, these findings reveal the temporal and mechanistic principles of PE connectivity, highlighting a potential role of PE contacts in establishing gene expression patterns in human development.
]]></description>
<dc:creator>Nocente, M. C.</dc:creator>
<dc:creator>Della Rosa, M.</dc:creator>
<dc:creator>Malcom, A. A.</dc:creator>
<dc:creator>Lister, G.</dc:creator>
<dc:creator>Savin, I.</dc:creator>
<dc:creator>Ray-Jones, H.</dc:creator>
<dc:creator>Elderkin, S.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Bendall, A.</dc:creator>
<dc:creator>Semprich, C. I.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Malysheva, V.</dc:creator>
<dc:creator>Rostovskaya, M.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:creator>Spivakov, M.</dc:creator>
<dc:date>2025-10-03</dc:date>
<dc:identifier>doi:10.1101/2025.10.02.679816</dc:identifier>
<dc:title><![CDATA[Poising and connectivity of emergent human developmental enhancers in the transition from naive to primed pluripotency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.02.680118v1?rss=1">
<title>
<![CDATA[
Lineage Tracing Reveals a Shared Cellular Origin for Supraclavicular Brown and Inguinal Beige Adipocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.02.680118v1?rss=1</link>
<description><![CDATA[
The metabolic importance of brown adipose tissue (BAT) has been recognized, but the origins of BAT, particularly supraclavicular BAT (scBAT), are less clear. Here, we traced the origin of scBAT to Mef2c-anterior heart field (AHF)-marked cells. Mef2c-AHF-marked cells isolated from scBAT can spontaneously differentiate into brown adipocytes, express mesenchymal stem cell markers, and can be isolated from the stromal-vascular fraction (SVF) of wild-type scBAT as [CD31-CD45-SCA-1+CD29+CD34-CD24-] (CD34-) cells. Mef2c-AHF-marked cells substantially overlap with Prrx1-marked cells in scBAT, which also contribute to beige adipocytes in inguinal white adipose tissue (iWAT) during development. Similarly, CD34- cells isolated from the SVF of iWAT can spontaneously differentiate into beige adipocytes in vitro. Using intersectional tracing, we revealed that common progenitors of Mef2c-AHF- and Prrx1-marked cells in scBAT emerge from the developing heart. Thus, these studies reveal a common origin for scBAT and cardiac tissue and a close lineage relationship between scBAT and beige adipocytes of the iWAT.
]]></description>
<dc:creator>Ran, Y.</dc:creator>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Shen, Y.-T.</dc:creator>
<dc:creator>Mo, Q.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Yalamanchili, H. K.</dc:creator>
<dc:creator>John, S.</dc:creator>
<dc:creator>Kogiso, M.</dc:creator>
<dc:creator>Gao, X.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Sinha, T.</dc:creator>
<dc:creator>Black, B.</dc:creator>
<dc:creator>Chen, M.-H.</dc:creator>
<dc:date>2025-10-03</dc:date>
<dc:identifier>doi:10.1101/2025.10.02.680118</dc:identifier>
<dc:title><![CDATA[Lineage Tracing Reveals a Shared Cellular Origin for Supraclavicular Brown and Inguinal Beige Adipocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.02.680148v1?rss=1">
<title>
<![CDATA[
Minor Cannabinoids CBD, CBG, CBN and CBC differentially modulate sensory neuron activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.02.680148v1?rss=1</link>
<description><![CDATA[
The use of minor cannabinoids has been advanced, in part, by the idea of providing relief from pain and inflammation without the burden of unwanted psychogenic effects associated with {Delta}9THC. In this regard, investigators have focused on the effects of minor cannabinoid activation / desensitization of peripheral sensory neurons on nociceptive signaling and/or peripheral inflammation. With a focus on peripheral nociception, four common minor cannabinoids: cannabidiol (CBD), cannabigerol (CBG), cannabinol (CBN) and cannabichromene (CBC) were studied in primary cultures of mouse Dorsal Root Ganglion (DRG) neurons. We queried if calcium responses induced by the four cannabinoids differed in potency of activation, neuronal size preference, and dose-response relationships. Additionally, we determined the dependence of CBD and CBN on key channel-receptors that are known to mediate pain and/or antinociception. Individually, CBD, CBG and CBC directed greater response magnitudes when compared to CBN. All four minor cannabinoids activated overlapping but distinct size populations of sensory neurons. CBD and CBG activated the widest range of DRG neuron sizes (smaller-larger) overlapping with smaller capsaicin-sensitive neurons. In contrast, CBN and CBC activated predominantly larger sensory neurons. CBD diverged from other minor cannabinoids in directing a linear dose-response profile whereas CBG and CBC directed sigmoidal dose-response profiles and CBN activated DRG neurons with an inverted U-shaped dose-response relationship. CBD-induced activation of DRG neurons was dependent on co-expression of the nociceptive channel TRPV1 plus cannabinoid receptor 1 (CB1R), whereas CBN-induced activation was independent of TRPV1. Overall, we observed that minor cannabinoids CBD, CBG, CBN and CBC differed in their activation of DRG neurons and directed unique activation properties across a diverse population of sensory neurons. Such differences underly the hypothesis that a combination (entourage) of complimentary minor cannabinoids can direct synergistic antinociceptive activity.
]]></description>
<dc:creator>Rabl, K.</dc:creator>
<dc:creator>Gruenke, L.</dc:creator>
<dc:creator>Banfal, A.</dc:creator>
<dc:creator>Eilers, H.</dc:creator>
<dc:creator>Hellman, J.</dc:creator>
<dc:creator>Schumacher, M. A.</dc:creator>
<dc:date>2025-10-03</dc:date>
<dc:identifier>doi:10.1101/2025.10.02.680148</dc:identifier>
<dc:title><![CDATA[Minor Cannabinoids CBD, CBG, CBN and CBC differentially modulate sensory neuron activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.03.680076v1?rss=1">
<title>
<![CDATA[
S1P induces bleb-based T cell motility via S1PR1-dependent activation of RhoA and WNK1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.03.680076v1?rss=1</link>
<description><![CDATA[
In vivo, the chemokine CCL19 and its receptor CCR7 control T cell retention in lymph nodes, while the lipid chemoattractant spingosine-1-phosphate (S1P) drives T cell egress from lymphoid organs. CCL19 is known to activate actin polymerization at the leading edge of migrating cells, generating a mode of motility driven by lamellipodial protrusions. In contrast, we showed recently that S1P induces a transient lamellipodial response, followed by pressure-driven bleb-based motility. Here, we elucidate the mechanisms controlling S1P responses in naive T cells. We show that S1P signals through S1PR1, with coupling through Gai. In contrast to CCR7, which signals through Gai to induce sustained Rac1 activation, S1PR1 engagement yields only weak and transient Rac1 activation; the dominant response is sustained activation of RhoA. This pathway, together with a pathway involving phospholipase C and myosin light chain kinase, results in phosphorylation of myosin regulatory light chain (MLC) and enhanced myosin contractility. Inhibition of mTORC2 blocks MLC phosphorylation, consistent with evidence that tension sensing by mTORC2 can couple Rac1 and RhoA signaling during leukocyte migration. Surprisingly, although RhoA pathway inhibitors blocked S1P-induced MLC phosphorylation and blebbing, they failed to block S1P-dependent chemotaxis. This led to the identification of a second arm of the S1P response: WNK1-dependent phosphorylation of SPAK1 and OSXR1, proteins that regulate ion channels and water influx. Partial WNK1 inhibition, together with inhibition of myosin contractility, was sufficient to block S1P-induced blebbing and chemotaxis, indicating that S1P-driven T cell migration involves coordinate activation of myosin contractility and water influx.

One sentence summaryS1P signals elicit sustained RhoA activation and water influx to drive bleb-based T cell motility.
]]></description>
<dc:creator>Staback Rodriguez, F.</dc:creator>
<dc:creator>De Belly, H.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Strickland, E.</dc:creator>
<dc:creator>Frazer, G. L.</dc:creator>
<dc:creator>Ojukwu, J.</dc:creator>
<dc:creator>Williamson, B. J.</dc:creator>
<dc:creator>Weiner, O. D.</dc:creator>
<dc:creator>Burkhardt, J. K.</dc:creator>
<dc:date>2025-10-04</dc:date>
<dc:identifier>doi:10.1101/2025.10.03.680076</dc:identifier>
<dc:title><![CDATA[S1P induces bleb-based T cell motility via S1PR1-dependent activation of RhoA and WNK1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.05.680493v1?rss=1">
<title>
<![CDATA[
Macrophage phagocytosis of Coccidioides promotes its differentiation into the parasitic form 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.05.680493v1?rss=1</link>
<description><![CDATA[
Coccidioides is an endemic fungus that is increasing in prevalence and can cause life threatening disease in otherwise immunocompetent people. In the environment the spores (arthroconidia) develop into hyphae, yet when they are inhaled by a mammalian host, they develop into a unique form called the spherule. The transition to spherule can be triggered in vitro with elevated temperatures and high CO2 levels, but the signals and host cells that might trigger Coccidioides spherulation in vivo are not known. We used live imaging to investigate how macrophages affect the fate of Coccidioides arthroconidia. Under tissue culture conditions, arthroconidia quickly developed into hyphae. The addition of macrophages promoted spherule development and delayed hyphal formation, indicating that innate immune cells can influence Coccidioides development into the pathogenic form. Exposure of arthroconidia to macrophage supernatants was not sufficient to stimulate spherulation, which was dependent on phagocytosis of arthroconidia by macrophages. Transcriptomics analysis of Coccidioides co-cultured with macrophages revealed a signature concordant with spherules grown in vitro and allowed the identification of a core set of spherule-specific transcripts. In addition, we identified Coccidioides transcripts with significantly higher abundance in the presence of macrophages compared to in vitro spherules, suggesting these factors may be needed to survive and thrive in the presence of innate immune cells. This work lays a foundation for uncovering host-pathogen signaling as well as Coccidioides factors that are critical for pathogenesis.
]]></description>
<dc:creator>Symington, J.</dc:creator>
<dc:creator>Dabholkar, A.</dc:creator>
<dc:creator>English, B.</dc:creator>
<dc:creator>Voorhies, M.</dc:creator>
<dc:creator>Sil, A.</dc:creator>
<dc:date>2025-10-05</dc:date>
<dc:identifier>doi:10.1101/2025.10.05.680493</dc:identifier>
<dc:title><![CDATA[Macrophage phagocytosis of Coccidioides promotes its differentiation into the parasitic form]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.04.680473v1?rss=1">
<title>
<![CDATA[
Multi-immersion Oblique Plane Microscope (miOPM): A reconfigurable platform for high-resolution Light-Sheet Fluorescence Microscopy. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.04.680473v1?rss=1</link>
<description><![CDATA[
Light-Sheet Fluorescence Microscopy (LSFM) enables gentle, rapid, and efficient volumetric imaging of biological specimens. Despite its potential, LSFM has fragmented into multiple variants, each narrowly optimized for specific samples or imaging regimes, limiting its broad applicability. To overcome this barrier, we introduce the multi-immersion oblique plane microscope (miOPM), an adaptable LSFM platform for high-resolution imaging ranging from subcellular dynamics to whole organisms and cleared tissues. miOPM uniquely supports seamless interchangeability of oil, water, and air objectives, and maintains diffraction limited performance across a refractive index range of 1.33 - 1.51. It is compatible with standard sample mounting, current clearing protocols and high-throughput 3D imaging. We leverage miOPM to image diverse biological specimens at multiple spatial scales. By greatly expanding the adaptability and ease of use of LSFM, miOPM is poised to democratize advanced three-dimensional imaging.
]]></description>
<dc:creator>Chen, B.</dc:creator>
<dc:creator>Millett-Sikking, A.</dc:creator>
<dc:creator>Galecki, S.</dc:creator>
<dc:creator>Daetwyler, S.</dc:creator>
<dc:creator>Jiou, J.</dc:creator>
<dc:creator>Monistrol, J.</dc:creator>
<dc:creator>Shen, Q.</dc:creator>
<dc:creator>Zhou, F. Y.</dc:creator>
<dc:creator>Lin, H.-Y.</dc:creator>
<dc:creator>Jenkins, E.</dc:creator>
<dc:creator>Stein, M.</dc:creator>
<dc:creator>Marlar-Pavey, M.</dc:creator>
<dc:creator>Sturm, G. Y.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:creator>Feng, B.</dc:creator>
<dc:creator>Diaz, U.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Shalizi, A.</dc:creator>
<dc:creator>Gillich, A.</dc:creator>
<dc:creator>Friedman, J. R.</dc:creator>
<dc:creator>Tomer, R. R.</dc:creator>
<dc:creator>Chang, B.-J.</dc:creator>
<dc:creator>Marshall, W.</dc:creator>
<dc:creator>Shahmoradian, S.</dc:creator>
<dc:creator>Dean, K. M.</dc:creator>
<dc:creator>Fiolka, R. P.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.04.680473</dc:identifier>
<dc:title><![CDATA[Multi-immersion Oblique Plane Microscope (miOPM): A reconfigurable platform for high-resolution Light-Sheet Fluorescence Microscopy.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.05.680386v1?rss=1">
<title>
<![CDATA[
Site-specific genome engineering of primary human natural killer cells for programmable anti-tumor function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.05.680386v1?rss=1</link>
<description><![CDATA[
Natural killer (NK) cells are emerging as a promising platform for engineered adoptive cell therapies. However, gene editing in NK cells remains challenging, and more effective strategies are needed. Here, we established a robust, feeder-free, and modular workflow for genome engineering in primary human NK cells, combining CRISPR/Cas9 with AAV6-mediated transgene delivery. Efficient site-specific transgene integration was achieved at various loci and can be coupled with concurrent disruption of the target locus in a single editing step. Furthermore, transgene expression was tunable according to the integration site and promoter. We applied this strategy to target a chimeric antigen receptor (CAR) transgene to a panel of inhibitory NK receptor loci, establishing a synergistic approach to enhance anti-tumor activity and facilitate the reliable comparison of CAR variants without expression bias. We identified TIGIT as an ideal locus that supports strong CAR expression and anti-tumor function. This genome engineering framework, which leverages multiple, complementary and precisely controlled genetic edits, can support the rational design of future NK-cell therapies tailored to overcome cell-intrinsic limitations and tumor-specific barriers.
]]></description>
<dc:creator>Allain, V.</dc:creator>
<dc:creator>Rothrock, A. G.</dc:creator>
<dc:creator>Bernard, P.-L.</dc:creator>
<dc:creator>Nyberg, W. A.</dc:creator>
<dc:creator>Talbot, A.</dc:creator>
<dc:creator>Muldoon, J. J.</dc:creator>
<dc:creator>Chung, J.-Y.</dc:creator>
<dc:creator>To, A.</dc:creator>
<dc:creator>Chang, C. R.</dc:creator>
<dc:creator>Kimmerly, G. R.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Tsao, T.</dc:creator>
<dc:creator>Mortazavi, Y.</dc:creator>
<dc:creator>Seo, J.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Shemesh, A.</dc:creator>
<dc:creator>Schmidt, R.</dc:creator>
<dc:creator>Ward, C. C.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Lanier, L. L.</dc:creator>
<dc:creator>Aguilar, O. A.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.05.680386</dc:identifier>
<dc:title><![CDATA[Site-specific genome engineering of primary human natural killer cells for programmable anti-tumor function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.05.680562v1?rss=1">
<title>
<![CDATA[
Base-editing a single missense mutation in A20 enhances CAR-T cell efficacy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.05.680562v1?rss=1</link>
<description><![CDATA[
T cell exhaustion limits the efficacy of cancer immunotherapies. Here, we performed genome-wide loss-of-function screening in repetitively stimulated human T cells and identified the mulitfunctional ubiquitin-modifying protein A20/TNFAIP3 as a major negative regulator of exhausted T cell persistence. Protein large language modeling, deep base-editing mutagenesis, and studies in immunocompetent mice with domain-specific inactivating mutations revealed A20s non-enzymatic M1 ubiquitin-binding zinc finger 7 (A20ZF7) motif as critical to suppression of anti-tumor immunity. A20ZF7-deficient CD8+ tumor-infiltrating lymphocytes (TILs) resisted terminal exhaustion and circumvented an unappreciated mechanism restraining perforin degranulation in terminally exhausted cells. Human chimeric antigen receptor (CAR)-T cells engineered via base-editing to inactivate A20ZF7 via a single missense mutation also resisted exhaustion, secreted more perforin and robustly suppressed cancer in vivo. These studies pinpoint A20ZF7 as a novel T cell checkpoint and reveal precision base-editing of missense mutations as an effective approach to enhance CAR-T cell therapy.
]]></description>
<dc:creator>Blaisdell, A.</dc:creator>
<dc:creator>Bachl, S.</dc:creator>
<dc:creator>Sandoval, L. R.</dc:creator>
<dc:creator>Ching, C.</dc:creator>
<dc:creator>Bowman, C. J.</dc:creator>
<dc:creator>Kale, N.</dc:creator>
<dc:creator>Prabandham, M.</dc:creator>
<dc:creator>Diolaiti, M.</dc:creator>
<dc:creator>Havig, C.</dc:creator>
<dc:creator>Advincula, R.</dc:creator>
<dc:creator>Lenci, N.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Yamashita, E.</dc:creator>
<dc:creator>Wang, C. H.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Achacoso, P.</dc:creator>
<dc:creator>Stibor, D.</dc:creator>
<dc:creator>Oynebraten, I.</dc:creator>
<dc:creator>Seo, J.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Malynn, B. A.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Carnevale, J.</dc:creator>
<dc:creator>Ma, A.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.05.680562</dc:identifier>
<dc:title><![CDATA[Base-editing a single missense mutation in A20 enhances CAR-T cell efficacy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.05.680571v1?rss=1">
<title>
<![CDATA[
A Dual Inhibitory Network in the Thalamic Reticular Nucleus Delineated by Pallidal and Intra-Reticular Inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.05.680571v1?rss=1</link>
<description><![CDATA[
Long described as an inhibitory "guardian of the gateway," the thalamic reticular nucleus (TRN) shapes which thalamic signals reach the cortex during attention, arousal, and sensory processing. However, how the inhibitory wiring within TRN supports this flexible gating--from modality-specific tuning to global control--remains poorly defined. Using cell type-specific optogenetic input mapping and whole-cell patch-clamp recordings in mice, we dissect inhibitory connectivity in TRN from two major GABAergic sources: the TRN itself and the external globus pallidus (GPe). All recorded TRN neurons received inhibition from the GPe, whereas a subset also received intra-TRN inhibition. Intra-TRN inhibition arose predominantly from somatostatin-expressing onto parvalbumin-expressing TRN neurons (SOM[-&gt;]PV), revealing subtype-specific connectivity recruited by thalamic excitation to form a feedforward motif. These findings delineate a dual inhibitory architecture: local intra-TRN circuits provide spatially selective inhibition, whereas pallidal inputs deliver diffuse inhibition. These complementary mechanisms may support both modality-specific gating and global state-dependent control of thalamic output.

Graphical AbstractThe thalamic reticular nucleus (TRN) integrates two distinct inhibitory motifs. Sparse, dendrite-targeting intra-TRN connections from parvalbumin- and somatostatin-expressing neurons provide structured, feedforward inhibition recruited by thalamocortical (TC) inputs. In parallel, robust and widespread inhibition from Npas1-expressing neurons of the external globus pallidus (GPe) supplies a strong external drive that can coordinate activity across the TRN. Together, these complementary motifs define a dual inhibitory architecture that may shape thalamocortical gating and highlight cellular entry points for therapeutic intervention.



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]]></description>
<dc:creator>Cho, F. S.</dc:creator>
<dc:creator>Huguenard, J. R.</dc:creator>
<dc:creator>Paz, J. T.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.05.680571</dc:identifier>
<dc:title><![CDATA[A Dual Inhibitory Network in the Thalamic Reticular Nucleus Delineated by Pallidal and Intra-Reticular Inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.06.680687v1?rss=1">
<title>
<![CDATA[
Consistent Induction of Broadly Neutralizing HIV Antibodies by a Novel Two-Step Mechanism Informs Immunogen Design 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.06.680687v1?rss=1</link>
<description><![CDATA[
A major obstacle confronting HIV-1 vaccine and cure research is the lack of an outbred animal model for rapid and consistent induction of broadly neutralizing antibodies (bNAbs). We designed an epitope-focused simian-human immunodeficiency virus (SHIV.5MUT) that elicited broad and potent V3-glycan-targeted antibodies within a year of infection in 14 of 22 macaques compared with 0 of 14 control animals. SHIV.5MUT elicited bNAbs by a novel two-step mechanism, inducing an initial wave of V1-directed antibodies that selected for Envs with shortened, hypoglycosylated V1 loops, which in turn primed V3-glycan bNAb precursors. Rhesus bNAbs were immunogenetically and structurally diverse, closely resembling human V3-glycan bNAbs. Env-bNAb coevolution revealed a diverse repertoire of bNAb precursors and the Env variants that matured them, yielding a molecular blueprint for vaccine design.
]]></description>
<dc:creator>Skelly, A. N.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Gavor, E.</dc:creator>
<dc:creator>Connell, A. J.</dc:creator>
<dc:creator>Kreider, E. F.</dc:creator>
<dc:creator>Marchitto, L.</dc:creator>
<dc:creator>Hogarty, M. P.</dc:creator>
<dc:creator>Newby, M. L.</dc:creator>
<dc:creator>Allen, J. D.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Ayyanathan, K.</dc:creator>
<dc:creator>Campion, M. S.</dc:creator>
<dc:creator>Winters, K.</dc:creator>
<dc:creator>Gordon, C. G.</dc:creator>
<dc:creator>Osbaldeston, R. A.</dc:creator>
<dc:creator>Akeley, M. J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Singh, A.</dc:creator>
<dc:creator>Cruickshank, K.</dc:creator>
<dc:creator>Park, Y.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Amereh, K.</dc:creator>
<dc:creator>Van Itallie, E.</dc:creator>
<dc:creator>Carey, J. W.</dc:creator>
<dc:creator>Albertus, A.</dc:creator>
<dc:creator>DeLaitsch, A. T.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Lituchy, M. G.</dc:creator>
<dc:creator>Walsh, A. A.</dc:creator>
<dc:creator>Morris, D. J.</dc:creator>
<dc:creator>Habib, R.</dc:creator>
<dc:creator>Bibollet-Ruche, F.</dc:creator>
<dc:creator>Mishra, N.</dc:creator>
<dc:creator>Avillion, G.</dc:creator>
<dc:creator>Koranda, N. S.</dc:creator>
<dc:creator>Plante, S. J.</dc:creator>
<dc:creator>Martella, C. L.</dc:creator>
<dc:creator>Lora, J.</dc:creator>
<dc:creator>Wang, E. J. D.</dc:creator>
<dc:creator>Lewis, M. G</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.06.680687</dc:identifier>
<dc:title><![CDATA[Consistent Induction of Broadly Neutralizing HIV Antibodies by a Novel Two-Step Mechanism Informs Immunogen Design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.06.680719v1?rss=1">
<title>
<![CDATA[
Impaired IFNγ responsiveness of lung monocyte-derived cells limits immunity to Mycobacterium tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.06.680719v1?rss=1</link>
<description><![CDATA[
Lung mononuclear phagocyte (MNP) subsets differ in their ability to restrict Mycobacterium tuberculosis (Mtb) during chronic infection, yet the mechanisms underlying this difference are not well defined. Here, we show that CD11clo monocyte-derived cells (MNC1), the subset of lung cells that is most permissive for Mtb viability during chronic infection, express lower levels of interferon-gamma (IFN{gamma}) signaling proteins, resulting in reduced responses to IFN{gamma} compared to alveolar macrophages (AM) and CD11chi MNC2. Moreover, type I IFN signaling suppresses IFN{gamma}-mediated MHC-II expression, impairing antigen-specific CD4 T cell activation by MNC1 cells. Importantly, prior immunity conferred by contained Mtb infection enhances IFN{gamma} responsiveness of monocyte-derived cells, reducing bacterial burdens in lungs and within MNC subsets. Our findings indicate that heterogeneous IFN{gamma} responsiveness is exploited by Mtb for persistence in vivo. Overcoming or bypassing impaired IFN{gamma} responsiveness may guide the development of more effective TB vaccines and host-directed therapies.
]]></description>
<dc:creator>Zheng, W.</dc:creator>
<dc:creator>Limberis, J. D.</dc:creator>
<dc:creator>Howard, Z. P.</dc:creator>
<dc:creator>Mohapatra, A.</dc:creator>
<dc:creator>Takagi, E.</dc:creator>
<dc:creator>Ernst, J. D.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.06.680719</dc:identifier>
<dc:title><![CDATA[Impaired IFNγ responsiveness of lung monocyte-derived cells limits immunity to Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.07.680994v1?rss=1">
<title>
<![CDATA[
Bayesian multi-state multi-condition modeling of a protein structure based on X-ray crystallography data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.07.680994v1?rss=1</link>
<description><![CDATA[
1An atomic structure model of a protein can be computed from a diffraction pattern of its crystal. While most crystallographic studies produce a single set of atomic coordinates, the billions of protein molecules in a crystal sample many conformational modes during data collection. As a result, a "multi-state" model that depicts these conformations could reproduce the X-ray data better than a single conformation, and thus likely be more accurate. Computing such a multistate model is challenging due to a lower data-to-parameter ratio than that for single-state modeling. To address this challenge, additional information could be considered, such as X-ray datasets collected for the same system under distinct experimental conditions (eg, temperature, ligands, mutations, and pressure). Here, we develop, benchmark, and illustrate MultiXray: Bayesian multi-state multi-condition modeling for X-ray crystallography. The input information is several X-ray datasets collected under distinct conditions and a molecular mechanics force field. The model consists of an independent coordinate set for each of several states and the weight of each state under each condition. A Bayesian posterior model density quantifies the match of the model with all X-ray datasets and the force field. A sample of models is drawn from the posterior model density using biased molecular dynamics (MD) simulations. We benchmark MultiXray on simulated CypA X-ray data. Using a second X-ray dataset improves the Rfree from 0.105 to 0.089. We then demonstrate MultiXray on experimental temperature-dependent data for SARS-CoV-2 Mpro. Using multiple X-ray datasets improves Rfree of the PDB-deposited structure from 0.253 to 0.237. MultiXray is implemented in our open-source Integrative Modeling Platform (IMP) software, relying on integration with Phenix, thus making it easily applicable to many studies.
]]></description>
<dc:creator>Hancock, M.</dc:creator>
<dc:creator>HOLTON, J. M.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Adams, P. D.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:date>2025-10-07</dc:date>
<dc:identifier>doi:10.1101/2025.10.07.680994</dc:identifier>
<dc:title><![CDATA[Bayesian multi-state multi-condition modeling of a protein structure based on X-ray crystallography data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.09.681498v1?rss=1">
<title>
<![CDATA[
Macrodomain ADP-ribose binding but not ADP-ribosylhydrolase activity is critical for chikungunya virus infection of Aedes mosquitoes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.09.681498v1?rss=1</link>
<description><![CDATA[
Viral macrodomains are promising antiviral targets that counteract host ADP-ribosylation-mediated antiviral responses in mammals. However, their role in dual-host viruses within the mosquito vector is largely unknown. Here, we investigated the role of the chikungunya virus (CHIKV) macrodomain by mutating the active site asparagine 24 (N24). In both mammalian and mosquito cell lines, these enzymes rapidly acquired compensatory mutations at aspartate 31 (D31). We show that while N24 mutations abolish ADP-ribosylhydrolase catalytic activity and reduce folding stability, ADP-ribose binding remains intact. Furthermore, the D31 compensatory mutations do not markedly rescue catalytic activity or folding stability. Structures of the compensatory mutant macrodomains suggest the importance of ADP-ribose binding, rather than ADP-ribosylhydrolase catalysis as the selective pressure driving their accumulation. In mammalian cells, viral mutants bearing the catalytic and compensatory mutations replicated less efficiently than wild-type virus in interferon-competent cell lines. However, their replication remained unaffected in mosquito cells. In Aedes mosquitoes, macrodomain mutations had disparate impacts, either reducing or enhancing infectivity and transmission depending on the specific mutation and viral lineage. These findings emphasize that viral macrodomain function is complex and host-dependent, highlighting the need for multi-host understanding to develop effective antivirals.
]]></description>
<dc:creator>Bardossy, E. S.</dc:creator>
<dc:creator>Bergmann, L.</dc:creator>
<dc:creator>Lacritick, A. H.</dc:creator>
<dc:creator>Nigg, J.</dc:creator>
<dc:creator>Correy, G. J.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Saleh, M. C.</dc:creator>
<dc:date>2025-10-09</dc:date>
<dc:identifier>doi:10.1101/2025.10.09.681498</dc:identifier>
<dc:title><![CDATA[Macrodomain ADP-ribose binding but not ADP-ribosylhydrolase activity is critical for chikungunya virus infection of Aedes mosquitoes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.08.681290v1?rss=1">
<title>
<![CDATA[
Ex Vivo Expanded Regulatory T Cells Inhibit AAA Progression by Limiting CD4+ and CD8+ T Cell Accumulation in Aortic Tissue 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.08.681290v1?rss=1</link>
<description><![CDATA[
BackgroundRegulatory T cells (Tregs) play a crucial role in the pathophysiology of abdominal aortic aneurysms (AAA), a chronic inflammatory condition with few treatment options for patients with early-stage disease. Treg therapy for AAA is potentially beneficial but its specific mechanism requires further investigation for clinical applications.

MethodsAfter identifying the critical role of T-cells in AAA using human and mouse AAA single-cell RNA sequencing data, we investigated the influence of Tregs on immune cell infiltration within mouse AAA--specifically CD3+ T cells--using congenic transfer of Thy1.1 allelic donor mice Tregs into AAA-induced wild-type C57BL/6J mice. AAA progression was quantified with ultrasound and image micrometry. Tissues obtained on postoperative days 7-42 were analyzed with flow cytometry, qRT-PCR, Verhoeff-van Gieson staining, hematoxylin-eosin staining, and immunohistochemistry.

ResultsCD3+ T cell population was profoundly elevated in the elastase induced AAA mouse model which was further used in the study. The AAA mice that received Treg cell therapy had less elastin degradation and aortic wall enlargement than their control counterparts. Donor Tregs were detected in draining lymph nodes even after five weeks, with characteristic expression of FOXP3 and CD25. Although donor Tregs were not detected in the aortic microenvironment, the pro-inflammatory cell population including CD4 and CD8 cells was reduced, compared to control mice.

ConclusionElevated T cell population aggravates inflammation and promotes AAA progression. Treg therapy impedes the recruitment of T cells into AAA tissue by colonizing the draining lymph nodes, thereby mitigating AAA progression. This study deepens our understanding of Treg stability, function, and potential as a promising therapy for early-stage aneurysms.
]]></description>
<dc:creator>Dasari, C.</dc:creator>
<dc:creator>Lopez, J. L.</dc:creator>
<dc:creator>Shawyan Jan, M.</dc:creator>
<dc:creator>Shaligram, S.</dc:creator>
<dc:creator>Lin, P.-Y.</dc:creator>
<dc:creator>Levy-Lambert, D.</dc:creator>
<dc:creator>Huang, A. G.</dc:creator>
<dc:creator>Lizama Valenzuela, C.</dc:creator>
<dc:creator>Hadley, P.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:creator>Oskowitz, A.</dc:creator>
<dc:date>2025-10-10</dc:date>
<dc:identifier>doi:10.1101/2025.10.08.681290</dc:identifier>
<dc:title><![CDATA[Ex Vivo Expanded Regulatory T Cells Inhibit AAA Progression by Limiting CD4+ and CD8+ T Cell Accumulation in Aortic Tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.09.681343v1?rss=1">
<title>
<![CDATA[
Metabolic Alterations driven by PFKFB3 upregulation confer Resistance to Trastuzumab in HER2-Positive Breast Cancer. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.09.681343v1?rss=1</link>
<description><![CDATA[
AimsResistance to anti-HER2 therapies, particularly trastuzumab, remains a major obstacle in the treatment of HER2-positive (HER2+) breast cancer. This study aims to uncover novel mechanisms driving trastuzumab resistance with a focus on the immune component, key mediator of trastuzumab efficacy.

MethodsWe developed an isogenic cell line-derived xenograft model to perform transcriptome-wide analyses of trastuzumab-sensitive and -resistant tumors. To validate key findings, we employed a 3D cancer-immune co-culture system capable of quantifying antibody-dependent cellular cytotoxicity (ADCC).

ResultsTranscriptomic profiling revealed how trastuzumab treatment shifts tumor transcriptomes, including changes that remodel the metabolic landscape and distinct gene signatures associated with resistance, notably the upregulation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3). Functional studies demonstrated that PFKFB3 promotes trastuzumab resistance by inducing metabolic rewiring and reducing ADCC. Silencing PFKFB3 restored immune-mediated cytotoxicity. Clinical dataset analyses confirmed that elevated PFKFB3 expression correlates with reduced overall and progression-free survival, and with incomplete pathological response to trastuzumab.

ConclusionsPFKFB3 upregulation drives metabolic adaptations that confer resistance to trastuzumab in HER2+ breast cancer. These findings highlight PFKFB3 as a promising therapeutic target to overcome resistance and improve patient outcomes.

HighlightsO_LIPFKFB3 upregulation promotes trastuzumab resistance in HER2+ breast cancer.
C_LIO_LITranscriptomic profiling reveals metabolic shifts linked to resistance.
C_LIO_LISilencing PFKFB3 restores ADCC and sensitizes cancer cells to trastuzumab.
C_LIO_LIHigh PFKFB3 expression correlates with poor survival and treatment response.
C_LIO_LIPFKFB3 is a potential therapeutic target to overcome trastuzumab resistance.
C_LI
]]></description>
<dc:creator>Vincken, R.</dc:creator>
<dc:creator>Pasco, S. T.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Bozal-Basterra, L.</dc:creator>
<dc:creator>van Ijcken, W. F. J.</dc:creator>
<dc:creator>Carracedo, A.</dc:creator>
<dc:creator>Huylebroeck, D.</dc:creator>
<dc:creator>Moasser, M. M.</dc:creator>
<dc:creator>Ruiz-Saenz, A.</dc:creator>
<dc:date>2025-10-10</dc:date>
<dc:identifier>doi:10.1101/2025.10.09.681343</dc:identifier>
<dc:title><![CDATA[Metabolic Alterations driven by PFKFB3 upregulation confer Resistance to Trastuzumab in HER2-Positive Breast Cancer.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.09.681482v1?rss=1">
<title>
<![CDATA[
Sensorimotor Theta Oscillations Coordinate Speech Movements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.09.681482v1?rss=1</link>
<description><![CDATA[
Fluent speech depends on precisely timed motor commands that coordinate rapid transitions between successive articulatory gestures. Using direct cortical recordings, we identified a prominent sensorimotor theta oscillation (6-10 Hz) that supports this coordination. During articulation, premotor speech circuits exhibited enhanced theta phase coherence, with elevated population activity near theta troughs. The oscillations frequency remained remarkably stable across varying speech rates and cognitive states, consistent with an intrinsically generated rhythm. Vocal-tract kinematics revealed pulse-like movements at 6-10 Hz, tightly coupled to cortical theta phase. At a mesoscopic scale, theta cycles structured sequential sensorimotor activations encoding articulatory gestures, with syllable identity optimally decodable following theta troughs. These findings identify theta oscillations as an intrinsic timing mechanism that coordinates the distributed and synergistic motor control underlying fluent speech.
]]></description>
<dc:creator>Norman, Y.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:date>2025-10-10</dc:date>
<dc:identifier>doi:10.1101/2025.10.09.681482</dc:identifier>
<dc:title><![CDATA[Sensorimotor Theta Oscillations Coordinate Speech Movements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.10.681741v1?rss=1">
<title>
<![CDATA[
IGF2BP3 remodels the microRNA targeting landscape in MLL-AF4 leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.10.681741v1?rss=1</link>
<description><![CDATA[
Insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3/I3) is a multi-domain RNA-binding protein required for MLL-AF4-driven leukemogenesis, but its mechanism of action remains enigmatic. We hypothesized from our previous work that I3 amplifies oncogenic gene expression by modulating RNA induced silencing complex (RISC) mRNA interactions. To test this, we performed miR-eCLIP of AGO2, the catalytic RISC subunit, in I3 knock-out (I3KO) as well as control B-cell acute lymphoblastic cell lines (B-ALL) and identified I3-dependent AGO2 binding sites on 111 3'UTRs. Analyzing chimeric miRNA-mRNA reads, we observed differential miRNA occupancy in the I3KO compared to control, including increased targeting by miR-181a, a regulator of leukocyte differentiation. Notably, miR-181a overexpression phenocopied I3 loss, implicating I3 in restricting miR-181a-mediated repression. Biochemical assays confirmed direct competition between I3 and AGO2-miRNA complexes for 3'-UTR binding. Taken together, our results provide a model for how I3 promotes leukemogenesis by antagonizing RISC-mediated repression of oncogenic mRNAs.
]]></description>
<dc:creator>Kabbani, L. E.</dc:creator>
<dc:creator>Kapoor, S.</dc:creator>
<dc:creator>Sharma, G. E.</dc:creator>
<dc:creator>Gutierrez, M.</dc:creator>
<dc:creator>Neeb, Z. T.</dc:creator>
<dc:creator>Jaiswal, A.</dc:creator>
<dc:creator>Ritter, A. J.</dc:creator>
<dc:creator>Katzman, S.</dc:creator>
<dc:creator>Feldman, J. E.</dc:creator>
<dc:creator>Rao, D. S.</dc:creator>
<dc:creator>Sanford, J. S.</dc:creator>
<dc:date>2025-10-13</dc:date>
<dc:identifier>doi:10.1101/2025.10.10.681741</dc:identifier>
<dc:title><![CDATA[IGF2BP3 remodels the microRNA targeting landscape in MLL-AF4 leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.12.681832v1?rss=1">
<title>
<![CDATA[
Transcriptional regulation of disease-relevant microglial activation programs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.12.681832v1?rss=1</link>
<description><![CDATA[
Microglia, the brains innate immune cells, can adopt a wide variety of activation states relevant to health and disease. Dysregulation of microglial activation occurs in numerous brain disorders, and driving or inhibiting specific states could be therapeutic. To discover regulators of microglial activation states, we conducted CRISPR interference screens in iPSC-derived microglia for inhibitors and activators of six microglial states. We identified transcriptional regulators for each of these states and characterized 31 regulators at the single-cell transcriptomic and cell-surface proteome level in two distinct iPSC-derived microglia models. Finally, we functionally characterized several regulators. STAT2 knockdown inhibits interferon response and lysosomal function. PRDM1 knockdown drives disease-associated and lipid-rich signatures and enhanced phagocytosis. DNMT1 knockdown results in widespread loss of methylation, activating negative regulators of interferon signaling. These findings provide a framework to direct microglial activation to selectively enrich microglial activation states, define their functional outputs, and inform future therapies.

HighlightsO_LICRISPRi screening reveals novel regulators of six microglia activation states
C_LIO_LIMulti-modal single-cell screens highlight differences between mRNA and protein level expression
C_LIO_LIiPSC-microglia models show different baseline distributions of activation states
C_LIO_LILoss of DNMT1 leads to widespread DNA demethylation, promoting some states but limiting the interferon-response state
C_LIO_LILoss of PRDM1 drives microglial disease-associated state
C_LI
]]></description>
<dc:creator>McQuade, A.</dc:creator>
<dc:creator>Mishra, R.</dc:creator>
<dc:creator>Hagan, V.</dc:creator>
<dc:creator>Liang, W.</dc:creator>
<dc:creator>Colias, P.</dc:creator>
<dc:creator>Castillo, V. C.</dc:creator>
<dc:creator>Lubin, J.</dc:creator>
<dc:creator>Haage, V.</dc:creator>
<dc:creator>Marshe, V.</dc:creator>
<dc:creator>Fujita, M.</dc:creator>
<dc:creator>Gomes, L.</dc:creator>
<dc:creator>Ta, T.</dc:creator>
<dc:creator>Teter, O.</dc:creator>
<dc:creator>Chasins, S. E.</dc:creator>
<dc:creator>De Jager, P. L.</dc:creator>
<dc:creator>Nunez, J. K.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:date>2025-10-13</dc:date>
<dc:identifier>doi:10.1101/2025.10.12.681832</dc:identifier>
<dc:title><![CDATA[Transcriptional regulation of disease-relevant microglial activation programs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.11.681833v1?rss=1">
<title>
<![CDATA[
Discovery of AVI-6451, a Potent and Selective Inhibitor of the SARS-CoV-2 ADP-Ribosylhydrolase Mac1 with Oral Efficacy in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.11.681833v1?rss=1</link>
<description><![CDATA[
The COVID-19 pandemic made plain the need for effective antivirals acting on novel antiviral targets, among which viral macrodomains have attracted considerable attention. We recently described AVI-4206 (1), a potent and selective inhibitor of the SARS-CoV-2 ADP-ribosylhydrolase Mac1 based on a 9H-pyrimido[4,5-b]indole core, the first Mac1 inhibitor to demonstrate antiviral efficacy in mouse models of SARS-CoV-2 infection, but requiring IP administration and frequent dosing. Herein we describe an extensive, structurally enabled medicinal chemistry effort to identify orally bioavailable Mac1 inhibitors by addressing permeability and efflux liabilities of 1 and many of its analogs. Multiple strategies were pursued to overcome these issues, including replacing a urea function to reduce hydrogen bond donor count. While heterocyclic urea mimetics could deliver analogs like AVI-6318 (3) with potencies and ADME profiles similar to 1, abrogation of the P-gp liability was finally achieved with entirely non-polar substituents in place of urea. Thus, AVI-6451 (4) is a potent Mac1 inhibitor lead with low intrinsic clearance, high oral bioavailability, and antiviral efficacy with once-daily oral administration in a mouse model of SARS-CoV-2 infection.
]]></description>
<dc:creator>Jaishankar, P.</dc:creator>
<dc:creator>Correy, G. J.</dc:creator>
<dc:creator>Matsui, Y.</dc:creator>
<dc:creator>Togo, T.</dc:creator>
<dc:creator>Rachman, M. M.</dc:creator>
<dc:creator>Stevens, M. G. V.</dc:creator>
<dc:creator>Hantz, E. R.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Diolaiti, M. E.</dc:creator>
<dc:creator>Montano, M.</dc:creator>
<dc:creator>Taha, T. Y.</dc:creator>
<dc:creator>Rosecrans, J.</dc:creator>
<dc:creator>Pampel, J.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Renslo, A. R.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.11.681833</dc:identifier>
<dc:title><![CDATA[Discovery of AVI-6451, a Potent and Selective Inhibitor of the SARS-CoV-2 ADP-Ribosylhydrolase Mac1 with Oral Efficacy in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.13.682155v1?rss=1">
<title>
<![CDATA[
CNPY4 is a Lipid-Binding Regulator of Sphingolipid Homeostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.13.682155v1?rss=1</link>
<description><![CDATA[
Sphingolipids function both as signaling molecules and as organizers of cell membranes, and their dysregulation has been linked to cancer, metabolic disorders, and neurodegeneration. A central node in the sphingolipid metabolism network is ceramide, which is converted into numerous derivatives, including sphingomyelin (SM). Through interactions with cholesterol, SM forms liquid-ordered microdomains that influence membrane organization and signaling. Previously, we reported that the Saposin-like (SAPLIP) protein Canopy4 (CNPY4) negatively regulates the levels of free cholesterol in the plasma membrane. Although SAPLIPs commonly regulate lipid metabolism through direct lipid interactions, CNPY4 does not bind cholesterol directly. Here, we show that CNPY4 interacts with multiple sphingolipids in vitro, including ceramide and SM, and with ceramide in cells. We also demonstrate that CNPY4 knockdown elevates SM levels at the plasma membrane and disrupts cellular localization and abundance of ceramide, suggesting that the levels and consequently the homeostatic distribution of these sphingolipids is under control of CNPY4. Although the most pronounced effect of CNPY4 loss is on the ceramide/SM conversion pathway, it additionally impacts the levels of over 150 cellular lipids and modulates neutral sphingomyelinase activity, consistent with secondary disruptions in sphingolipid homeostasis. Collectively, our findings point to CNPY4 as a sphingolipid chaperone that regulates the abundance and localization of these lipids, modulating in turn cholesterol homeostasis and cellular signaling.
]]></description>
<dc:creator>Paul, M. D.</dc:creator>
<dc:creator>Zbodakova, O.</dc:creator>
<dc:creator>Glenn, I. S.</dc:creator>
<dc:creator>Stotland, A.</dc:creator>
<dc:creator>Klein, O. D.</dc:creator>
<dc:creator>Jura, N.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.13.682155</dc:identifier>
<dc:title><![CDATA[CNPY4 is a Lipid-Binding Regulator of Sphingolipid Homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.14.682265v1?rss=1">
<title>
<![CDATA[
Single-cell transcriptomic profiling of bam mutant tumor reveals germline heterogeneity and gcrf1 as a modulator in Drosophila germ cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.14.682265v1?rss=1</link>
<description><![CDATA[
The bam mutant ovary of Drosophila represents a classic tumor model caused by germline stem cell (GSC) differentiation defects. To date, its molecular and genetic features have rarely been characterized in detail at the single-cell resolution. Here, we performed single-cell RNA sequencing (scRNA-seq) to comprehensively delineate the transcriptomic landscape and identify distinct germline cell types in bam mutant ovaries by using in situ hybridization. Differentially expressed gene analysis and PAGA plots reveal different transcriptional profiles and developmental relationships in ovarian cells. Based on the expression pattern of eggpl, a useful marker for undifferentiated germ cell identity, and morphological differences in bam mutant ovarioles, two potentially distinct germ cell states are distinguished. Comparative single-cell analysis reveals the potential regulatory network and cellular communication in subclusters of undifferentiated germ cells, and contributes to the identification of gcrf1 as a novel marker gene for female GSC, which involves in the regulation of early germ cell proliferation and Drosophila fertility. Collectively, our study not only provides insights into tumorigenesis caused by GSC differentiation defects but also offers a valuable transcriptomic resource that can be mined for the reproductive features of bam mutant tumors by community.
]]></description>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Zeng, Y.</dc:creator>
<dc:creator>Nystul, T. G.</dc:creator>
<dc:creator>Zhong, G.</dc:creator>
<dc:date>2025-10-15</dc:date>
<dc:identifier>doi:10.1101/2025.10.14.682265</dc:identifier>
<dc:title><![CDATA[Single-cell transcriptomic profiling of bam mutant tumor reveals germline heterogeneity and gcrf1 as a modulator in Drosophila germ cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.14.682425v1?rss=1">
<title>
<![CDATA[
New proteomic biomarkers identified in plasma extracellular vesicles in sarcoidosis: a case-control matched study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.14.682425v1?rss=1</link>
<description><![CDATA[
BACKGROUNDSarcoidosis is a heterogeneous disease with unknown mechanisms, nonspecific therapies, and multiple etiologies. The role of blood extracellular vesicles (EVs) in the diagnosis and pathogenesis of sarcoidosis remains obscure. AIMS/OBJECTIVES. This study aims to test the hypothesis that the EV proteins in the blood can serve as phenotypic biomarkers of sarcoidosis. METHODS. We combined EV proteomics with machine learning algorithms to identify and prioritize biomarkers, enrich their functions, and cluster networks in case-control matched ACCESS patients. RESULTS. In total, 278 plasma EV proteins were significantly upregulated or downregulated in 40 sarcoidosis patients compared with 40 matched healthy controls. We identified 97 proteins that could serve as biomarkers with an AUC > 0.75. Of these, the AUC was > 0.90 for 13 proteins. 62 differentially expressed EV proteins strongly correlated with 20 clinical variables of severity, chest X-ray findings, and/or laboratory results. Functional annotation and network analysis suggest that these differentially expressed proteins regulate endocytosis, host responses to external stimuli, and transcription processes. Moreover, the top three ranked pathways were clathrin-mediated endocytosis, Hsp90 chaperone cycle, and spliceosome. CONCLUSIONS. This study demonstrates that plasma EV proteins can serve as biomarkers of various clinical phenotypes of the disease.

At a Glance CommentaryCurrent Scientific Knowledge on the Subject: Sarcoidosis is a heterogeneous condition affecting multiple organs. The role of blood extracellular vesicles in the diagnosis and pathogenesis of this condition remains unknown.

What This Study Adds to the Field: We identified differentially expressed proteins in plasma EVs by combining proteomics and machine learning algorithms. Top-ranked proteins can serve as diagnostic biomarkers and potential mechanisms for the development of sarcoidosis.
]]></description>
<dc:creator>Xi, N. M.</dc:creator>
<dc:creator>Gabby, L.</dc:creator>
<dc:creator>Zhao, R.</dc:creator>
<dc:creator>Vanarsa, K.</dc:creator>
<dc:creator>Qiao, M.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Mohan, C.</dc:creator>
<dc:creator>Judson, M. A.</dc:creator>
<dc:creator>Koth, L. L.</dc:creator>
<dc:creator>JI, H.</dc:creator>
<dc:date>2025-10-16</dc:date>
<dc:identifier>doi:10.1101/2025.10.14.682425</dc:identifier>
<dc:title><![CDATA[New proteomic biomarkers identified in plasma extracellular vesicles in sarcoidosis: a case-control matched study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.15.682722v1?rss=1">
<title>
<![CDATA[
Fine-Tuning Protein Language Models on Human Spatial Constraint Yields State-of-the-Art Variant Effect Prediction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.15.682722v1?rss=1</link>
<description><![CDATA[
Protein language models (PLMs) achieve state-of-the-art performance in predicting effects of missense variants, yet they do not explicitly consider variation within the human population. Here, we introduce Human Spatial Constraint (HuSC), a framework for quantifying intraspecies constraint on missense variants that integrates population-scale human genetic variation with 3D protein structures. We then fine-tune PLMs on HuSC scores. HuSC models the expected frequency of missense variation under neutral evolution and compares it to observed variation, accounting for both variation in mutational processes and 3D structural context. HuSC outperforms traditional inter- and intraspecies conservation metrics in predicting pathogenic variants. By focusing on intraspecies variation, HuSC reveals protein sites under human-specific constraint that cannot be captured by interspecies models. Integrating this intraspecies perspective into PLMs by fine-tuning on HuSC scores improves the prediction of variant fitness from deep mutational scans across diverse taxa and functional assay types. The improvement after fine-tuning comes largely from reducing bias toward wild-type sequences in regions that tolerate variation. Together, these results demonstrate that combining intraspecies constraint with cross-species PLMs improves their performance in variant-effect interpretation.
]]></description>
<dc:creator>Bajracharya, G.</dc:creator>
<dc:creator>Capra, J. A.</dc:creator>
<dc:date>2025-10-16</dc:date>
<dc:identifier>doi:10.1101/2025.10.15.682722</dc:identifier>
<dc:title><![CDATA[Fine-Tuning Protein Language Models on Human Spatial Constraint Yields State-of-the-Art Variant Effect Prediction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.16.682879v1?rss=1">
<title>
<![CDATA[
In silico reconstruction of primary and metastatic tumor architecture using GIS-augmented spatial transcriptomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.16.682879v1?rss=1</link>
<description><![CDATA[
The tumor microenvironment (TME) comprises different cell populations that interact, contributing to tumor heterogeneity and therapy response. Spatial transcriptomics offers valuable insights into transcriptional complexity and heterogeneity of the TME. We established Geographic Information System (GIS)-augmented In-Silico Reconstruction of Tumor Architecture (GIS-ROTA), a biologically informed analytic framework that integrates pathway or cell type-based enrichment analysis with local Morans I to uncover functional spatial domains. In our Visium dataset of primary and metastatic estrogen receptor-positive breast tumor samples, GIS-ROTA revealed extensive co-localization of estrogen response with metabolic pathway gene sets and mutual exclusivity with metastasis-related and specific immune-related pathway gene sets. The novelty of our approach lies in considering biological functions prior to identifying any spatial domains, providing direct interpretability and minimizing the subjectivity of interpreting clusters observed from conventional analytic methods. Overall, our GIS-ROTA framework integrates biological knowledge first, yielding spatial patterns with functional relevance and enabling identification of novel targets for development of therapeutic strategies.
]]></description>
<dc:creator>Yoo, J. Y.</dc:creator>
<dc:creator>Akter, S.</dc:creator>
<dc:creator>Zuo, Q.</dc:creator>
<dc:creator>Kazemi, A.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Goel, M.</dc:creator>
<dc:creator>Lam, A.</dc:creator>
<dc:creator>Dutta, D.</dc:creator>
<dc:creator>Barnick, B.</dc:creator>
<dc:creator>Pekmezci, M.</dc:creator>
<dc:creator>Grosse Perdekamp, M.</dc:creator>
<dc:creator>Soliman, A.</dc:creator>
<dc:creator>Madak-Erdogan, Z.</dc:creator>
<dc:date>2025-10-17</dc:date>
<dc:identifier>doi:10.1101/2025.10.16.682879</dc:identifier>
<dc:title><![CDATA[In silico reconstruction of primary and metastatic tumor architecture using GIS-augmented spatial transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.17.683123v1?rss=1">
<title>
<![CDATA[
Antibody Reveals Conformational Latch Controlling Herpesvirus Proteases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.17.683123v1?rss=1</link>
<description><![CDATA[
Human herpesviruses (HHVs) are widespread pathogens that cause severe disease. Their replication depends on the HHV protease (HHV Pr), an enzyme essential for capsid maturation. Because HHV Pr must dimerize to become catalytically active, disrupting dimer formation is a promising strategy for antiviral therapeutic development. We isolated a conformationally selective antibody inhibitor, Fab5, from a fully human naive Fab-phage library that recognizes monomeric human cytomegalovirus protease (HCMV Pr). A 2.6 [A] cryoelectron microscopy (cryo-EM) structure of the Fab5-HCMV Pr complex revealed that Fab5 binds a flexible loop distal from the active site and dimer interface which we call the latch loop. In HCMV Pr dimers, this loop secures the C-terminal tail to the protein core. Structure-guided mutagenesis confirmed that the latch loop is essential for HCMV Pr dimerization and activity. This loop is structurally conserved across all HHV Prs, and we show its functional role in Kaposis Sarcoma-associated herpesvirus (KSHV) Pr as well. The latch loop plays a mechanistic role in the conformational transition required for HHV Pr activity, and it forms a cryptic site that presents a new avenue for future allosteric inhibitor development.
]]></description>
<dc:creator>Zimanyi, M.</dc:creator>
<dc:creator>Hulce, K. R.</dc:creator>
<dc:creator>Bohn, M. F.</dc:creator>
<dc:creator>Norman, J.</dc:creator>
<dc:creator>Rohweder, P. J.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Craik, C.</dc:creator>
<dc:date>2025-10-17</dc:date>
<dc:identifier>doi:10.1101/2025.10.17.683123</dc:identifier>
<dc:title><![CDATA[Antibody Reveals Conformational Latch Controlling Herpesvirus Proteases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.18.683228v1?rss=1">
<title>
<![CDATA[
All-atom protein design via SE(3) flow matching with ProteinZen 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.18.683228v1?rss=1</link>
<description><![CDATA[
The advent of generative models of protein structure has greatly accelerated our ability to perform de novo protein design, especially concerning design at coarser physical scales such as backbone generation and protein binder design. How-ever, the design of precise placements at atomic scales remains a challenge for existing design methods. One avenue towards higher fidelity atomic-scale design is via generative models with full atomic resolution, but is complicated by the intricacies of simultaneously designing both discrete protein sequence and continuous atomic positionings. In this work we propose a framework to capture this interplay by decomposing residues into collections of oriented rigid bodies, allowing us to apply SE(3) flow-matching for all-atom protein structure generation. Our method, ProteinZen1, generates designs with high sequence-structure consistency while retaining competitive diversity and novelty on both unconditional and conditional generation tasks. We demonstrate competitive performance for unconditional monomer design and state-of-the-art performance on various forms of motif scaffolding, including full-atom motif scaffolding and motif scaffolding without specifying motif segment spacing or relative sequence order.
]]></description>
<dc:creator>Li, A. J.</dc:creator>
<dc:creator>Kortemme, T.</dc:creator>
<dc:date>2025-10-18</dc:date>
<dc:identifier>doi:10.1101/2025.10.18.683228</dc:identifier>
<dc:title><![CDATA[All-atom protein design via SE(3) flow matching with ProteinZen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.18.683207v1?rss=1">
<title>
<![CDATA[
The neural basis of emotional generalization in empathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.18.683207v1?rss=1</link>
<description><![CDATA[
The essence of empathy is generalization of emotion across persons. Here, we leverage recent theoretical advances in the neuroscience of generalization to help us understand empathy. We measured brain activity in human neurosurgical patients performing two tasks, one focused on identifying their own emotional response and one identifying emotional responses in others. We quantified the representational geometry of local field potential (LFP) high-gamma activity in four regions: the medial temporal lobe, anterior cingulate cortex, orbitofrontal cortex, and insula. We found encoding of both self- and other-emotions in all four regions, but codes for emotion and person are disentangled (that is, factorized) in the insula, but not the other regions. This factorized representation allows for cross-person generalization of emotion in a way that tangled (non-factorized) representations do not. Together, these results support the hypothesis that the insula uniquely contributes to social mirroring processes by which we understand emotions across individuals.
]]></description>
<dc:creator>Hayden, B.</dc:creator>
<dc:creator>Allawalla, A.</dc:creator>
<dc:creator>Adkinson, J.</dc:creator>
<dc:creator>Fan, X.</dc:creator>
<dc:creator>Franch, M.</dc:creator>
<dc:creator>Gates, V.</dc:creator>
<dc:creator>Mathura, R.</dc:creator>
<dc:creator>Pascuzzi, B.</dc:creator>
<dc:creator>Mocchi, M.</dc:creator>
<dc:creator>Myers, J.</dc:creator>
<dc:creator>Pulapaka, S.</dc:creator>
<dc:creator>Banks, G.</dc:creator>
<dc:creator>Bartoli, E.</dc:creator>
<dc:creator>Goodman, W.</dc:creator>
<dc:creator>Mathew, S.</dc:creator>
<dc:creator>Pitkow, X.</dc:creator>
<dc:creator>Pouratian, N.</dc:creator>
<dc:creator>Provenza, N.</dc:creator>
<dc:creator>Shofty, B.</dc:creator>
<dc:creator>Watrous, A.</dc:creator>
<dc:creator>Bijanki, K.</dc:creator>
<dc:creator>Sheth, S.</dc:creator>
<dc:creator>Yoo, S. B. M.</dc:creator>
<dc:date>2025-10-19</dc:date>
<dc:identifier>doi:10.1101/2025.10.18.683207</dc:identifier>
<dc:title><![CDATA[The neural basis of emotional generalization in empathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.18.683238v1?rss=1">
<title>
<![CDATA[
ANTIPODE Provides a Global View of Cell Type Homology and Transcriptomic Divergence in the Developing Mammalian Brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.18.683238v1?rss=1</link>
<description><![CDATA[
Diverse neurons and glia are generated in conserved spatial and temporal sequences during mammalian brain development. Divergence in gene regulatory networks can alter brain composition, scaling, timing, and function. However, resolving the identity, extent, and principles of gene regulatory divergence requires cellular-resolution surveys spanning brain regions and species and improved methods for defining cell type homologies. Here, we present ANTIPODE, a deep-learning variational inference framework that simultaneously integrates single-cell datasets, identifies homologous cell types, and parcellates differential expression across cell types, modules, and covariance. Applying ANTIPODE to a census of the whole developing macaque brain and a meta-atlas of human, macaque, and mouse brain development, we find broad conservation of initial neuron classes but widespread regulatory divergence within homologous types, shaped by genomic context, cell lineage, and developmental timing. Together, ANTIPODE provides a formalized and interpretable framework for cross-species single-cell analysis and reveals principles of gene regulatory divergence in mammalian brain evolution.
]]></description>
<dc:creator>Schmitz, M. T.</dc:creator>
<dc:creator>Ding, J. W.</dc:creator>
<dc:creator>Nolbrant, S.</dc:creator>
<dc:creator>McMullen, R.</dc:creator>
<dc:creator>Kim, C. N.</dc:creator>
<dc:creator>Pavlovic, B. J.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Bakken, T. E.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:date>2025-10-19</dc:date>
<dc:identifier>doi:10.1101/2025.10.18.683238</dc:identifier>
<dc:title><![CDATA[ANTIPODE Provides a Global View of Cell Type Homology and Transcriptomic Divergence in the Developing Mammalian Brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.19.683305v1?rss=1">
<title>
<![CDATA[
Autoregulation of the MET Receptor Tyrosine Kinase by its Intracellular Juxtamembrane Domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.19.683305v1?rss=1</link>
<description><![CDATA[
Receptor Tyrosine Kinases (RTKs) are single-pass transmembrane receptors whose activation is tightly regulated by intra-domain interactions within both their extracellular and intracellular regions. The intracellular juxtamembrane domain, which links the transmembrane and kinase domains, often plays a critical role in modulating kinase activity. The MET receptor, activated by Hepatocyte Growth Factor (HGF), requires precise regulation to support normal development and wound healing, but becomes a potent oncogene when overexpressed or mutated. A common oncogenic lesion in MET, caused by exon 14 skipping, leads to partial deletion of its unusually long intracellular juxtamembrane domain and is frequently detected in non-small cell lung cancer (NSCLC), as well as pancreatic, liver and brain cancers. Despite its length and abundance of post-translational modifications, the functional role of the MET juxtamembrane domain has remained poorly understood. We have uncovered that this segment regulates the kinetics of MET kinase activation. Specifically, we found that a membrane-proximal, N-terminal region of the juxtamembrane domain accelerates activation loop phosphorylation promoting kinase transition to an active state. This regulation does not depend on the oligomeric state of MET but likely acts allosterically to enhance autophosphorylation of the kinase domain. Notably, this function is absent in the closely related MST1R/RON RTK, suggesting it is a unique feature of the MET receptor. Together, these findings uncover a previously unrecognized layer of MET regulation with potential implications for the development of selective therapies targeting MET-driven cancers.
]]></description>
<dc:creator>Linossi, E. M.</dc:creator>
<dc:creator>Espinoza, C. A.</dc:creator>
<dc:creator>Estevam, G. O.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Jura, N.</dc:creator>
<dc:date>2025-10-19</dc:date>
<dc:identifier>doi:10.1101/2025.10.19.683305</dc:identifier>
<dc:title><![CDATA[Autoregulation of the MET Receptor Tyrosine Kinase by its Intracellular Juxtamembrane Domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.19.683269v1?rss=1">
<title>
<![CDATA[
Self-supervised learning enables robust microbiome predictions in data-limited and cross-cohort settings 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.19.683269v1?rss=1</link>
<description><![CDATA[
The gut microbiome plays a crucial role in human health, but machine learning applications in this field face significant challenges, including limited labeled data availability, high dimensionality, and batch effects across different cohorts. To address these limitations, we developed representation learning models for gut microbiome metagenomic data, drawing inspiration from foundation models approaches based on self-supervised and transfer learning principles. By leveraging a large collection of 85,364 metagenomic samples, we implemented multiple self-supervised learning methods, including masked autoencoders with varying masking rates and adapted single-cell RNA sequencing models (scVI and scGPT), to generate embeddings from bacterial abundance profiles. These learned representations demonstrated significant advantages over raw bacterial abundances in two key scenarios: first, when training predictive models with very limited labeled data, improving prediction performance for age (r = 0.14 vs. 0.06), BMI (r = 0.16 vs. 0.11), visceral fat mass (r = 0.25 vs. 0.18), and drug usage classification (PR-AUC = 0.81 vs. 0.73); and second, when generalizing predictions across different cohorts, consistently outperforming models based on raw abundances in cross-dataset evaluation. Our approach provides a valuable framework for leveraging self-supervised representation learning to overcome the data limitations inherent in microbiome research, potentially enabling more robust and generalizable machine learning applications in this field.
]]></description>
<dc:creator>Zahavi, L.</dc:creator>
<dc:creator>Levine, Z.</dc:creator>
<dc:creator>Godneva, A.</dc:creator>
<dc:creator>Dubinkina, V.</dc:creator>
<dc:creator>Dhir, R.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Weinberger, A.</dc:creator>
<dc:creator>Segal, E.</dc:creator>
<dc:date>2025-10-20</dc:date>
<dc:identifier>doi:10.1101/2025.10.19.683269</dc:identifier>
<dc:title><![CDATA[Self-supervised learning enables robust microbiome predictions in data-limited and cross-cohort settings]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.20.683419v1?rss=1">
<title>
<![CDATA[
Oncogenic and immunomodulatory functions of SUV420H1 in HPV-negative head and neck squamous cell carcinoma. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.20.683419v1?rss=1</link>
<description><![CDATA[
Despite the advent of immunotherapy, human-papilloma-virus (HPV)-negative head and neck squamous cell carcinoma (HNSCC) carries a high morbidity and mortality rate, thus novel therapies are urgently needed. Suppressor Of Variegation 4-20 Homolog 1 (SUV420H1) is a protein lysine methyltransferase that writes H4K20me3. Approximately 35% of HPV-negative HNSCC tumors carry gains/amplifications of SUV420H1. Gene Set Enrichment Analysis (GSEA) showed enrichment of proliferation-, epithelial-mesenchymal transition (EMT)- and immune-response pathways in SUV420H1-overexpressing HPV-negative HNSCC tumors. Depletion of SUV420H1 led to decreased proliferation, cell cycling and invasion in human HPV-negative HNSCC cell lines, while enzymatic inhibition decreased the invasive capacity but not the proliferation and cell cycling of HPV-negative HNSCC cell lines, supporting the presence of catalytically-independent and -dependent functions of SUV420H1. In a syngeneic mouse model of mouse oral carcinoma 1 tumors (MOC1), Suv420h1 knockout (KO) in MOC1 cancer cells halted tumor growth and synergized with anti-PD-1 therapy. In the tumor immune microenvironment (TIME) of Suv420h1 KO MOC1 tumors, a significant increase in the macrophage compartment with a concurrent decrease in the protumorigenic granulocytic myeloid derived suppressor cells (gMDSCs) was observed. Genome-wide mapping of SUV420H1-mediated H4K20me3 revealed enrichment of EMT-, IFN-response, and myeloid-attracting chemokines. This work provides rationale for the depletion/degradation of SUV420H1 as a novel therapeutic strategy to hinder proliferation and invasion, and to impart sensitization to anti-PD-1 immunotherapy for patients with HPV-negative HNSCC tumors with SUV420H1 gain/amplification.
]]></description>
<dc:creator>Moshiri, A.</dc:creator>
<dc:creator>Luff, M.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Jiang, M.</dc:creator>
<dc:creator>Dar, M. S.</dc:creator>
<dc:creator>Patel, M.</dc:creator>
<dc:creator>McKinnon, K.</dc:creator>
<dc:creator>Edmondson, E.</dc:creator>
<dc:creator>Akhtar, J.</dc:creator>
<dc:creator>Cheng, H.</dc:creator>
<dc:creator>Saloura, V.</dc:creator>
<dc:date>2025-10-20</dc:date>
<dc:identifier>doi:10.1101/2025.10.20.683419</dc:identifier>
<dc:title><![CDATA[Oncogenic and immunomodulatory functions of SUV420H1 in HPV-negative head and neck squamous cell carcinoma.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.21.683747v1?rss=1">
<title>
<![CDATA[
mRNA delivery of genetically encoded mosaic-8 pan-sarbecovirus RBD vaccines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.21.683747v1?rss=1</link>
<description><![CDATA[
Effective pan-sarbecovirus vaccines could prevent future zoonotic spillovers of SARS-like betacoronaviruses. We previously developed protein-based mosaic-8 nanoparticles displaying eight diverse sarbecovirus RBDs, either individually (mosaic-8 RBD-NPs) or as two "quartets" of four tandemly-arranged RBDs (dual-quartet RBD-NPs), which elicited broadly cross-reactive antibodies but require multi-component manufacturing. Here, we address scalability challenges by extending the mosaic-8 concept to mRNA by encoding membrane-bound RBD quartets as dual-quartet RBD-mRNA and dual-quartet RBD-EABR-mRNA, the latter leveraging ESCRT- and ALIX-binding region (EABR) technology for immunogen display on cell surfaces and secreted virus-like particles. Compared with protein-based mosaic-8 immunogens, mRNA-encoded mosaic-8 vaccines induced equivalent or enhanced antibody breadth, neutralization potencies, T-cell responses, and targeting of conserved RBD epitopes. In addition, mRNA-encoded mosaic-8 vaccines elicited more balanced IgG subclass profiles and increased Fc{gamma} receptor-binding IgGs, consistent with potentially superior Fc effector functions. These findings demonstrate successful translation of mosaic-8 RBD-NPs into mRNA/EABR-mRNA vaccines, enabling scalable manufacturing and improving protection against future sarbecovirus outbreaks. Finally, our newly developed technique, Systems Serology-Polyclonal Epitope Mapping (SySPEM), revealed distinct IgG-subclass-specific epitope signatures across mRNA, EABR-mRNA, and protein vaccines, demonstrating that the mode of antigen display can shape epitope recognition.

SummaryWe translated a pan-sarbecovirus RBD vaccine from protein nanoparticles to scalable mRNA and EABR-mRNA platforms encoding RBD quartets. Compared with protein-based immunogens, mRNA-based vaccines matched or improved antibody breadth, T-cell responses, Fc functionality, and conserved epitope targeting. A newly-developed Systems Serology-Polyclonal Epitope Mapping (SySPEM) technique revealed that antigen presentation modality shapes IgG subclass-specific epitope recognition.
]]></description>
<dc:creator>Cohen, A. A.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Manasyan, L.</dc:creator>
<dc:creator>Madireddy, I.</dc:creator>
<dc:creator>Fils, A.-C. I. P.</dc:creator>
<dc:creator>Dam, K.-M. A.</dc:creator>
<dc:creator>Stober, H. E.</dc:creator>
<dc:creator>Hills, R. A.</dc:creator>
<dc:creator>Moon, W. J.</dc:creator>
<dc:creator>Lin, P. J. C.</dc:creator>
<dc:creator>Howarth, M. R.</dc:creator>
<dc:creator>Hoffmann, M. A. G.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2025-10-22</dc:date>
<dc:identifier>doi:10.1101/2025.10.21.683747</dc:identifier>
<dc:title><![CDATA[mRNA delivery of genetically encoded mosaic-8 pan-sarbecovirus RBD vaccines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.21.683815v1?rss=1">
<title>
<![CDATA[
ADAPT-M: A workflow for rapid, quantitative in vitro measurements of enriched protein libraries 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.21.683815v1?rss=1</link>
<description><![CDATA[
Protein-protein interactions underpin most cellular interactions, and engineered binders present powerful tools for probing biology and developing novel therapeutics. One bottleneck in binder generation is the scalable, quantitative characterization of these interactions. We present ADAPT-M (Affinity Determination by Adaptation of ProTein binders for Microfluidics), a streamlined workflow that connects yeast surface display (YSD) with in vitro affinity and kinetic measurements using the high-throughput STAMMPPING microfluidic platform. ADAPT-M quantifies Kds and dissociation kinetic parameters for hundreds of enriched protein variants in under one week without requiring hands-on protein purification. We applied ADAPT-M to a computationally designed library targeting the SARS-CoV-2 Omicron BA.1 receptor binding domain, successfully recovering and measuring Kds for most highly enriched YSD variants. Measurements correlate strongly with biolayer interferometry and yeast titration assays. ADAPT-M further enabled selection of lead candidates for structural and mutational analysis, which revealed designed paratopes were preserved despite binding to off-target epitopes. By bridging YSD screening and in vitro validation, ADAPT-M accelerates protein binder discovery and supports data-driven protein engineering.
]]></description>
<dc:creator>Perez, C. P.</dc:creator>
<dc:creator>DelRosso, N. V.</dc:creator>
<dc:creator>Noland, C. L.</dc:creator>
<dc:creator>Parekh, U.</dc:creator>
<dc:creator>Choe, C. A.</dc:creator>
<dc:creator>Eguchi, R. R.</dc:creator>
<dc:creator>Wen, Q.</dc:creator>
<dc:creator>Fordyce, P. M.</dc:creator>
<dc:creator>Huang, P.</dc:creator>
<dc:date>2025-10-22</dc:date>
<dc:identifier>doi:10.1101/2025.10.21.683815</dc:identifier>
<dc:title><![CDATA[ADAPT-M: A workflow for rapid, quantitative in vitro measurements of enriched protein libraries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.22.683967v1?rss=1">
<title>
<![CDATA[
Reverse transcribed ssDNA derepresses translation of a retron antiviral protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.22.683967v1?rss=1</link>
<description><![CDATA[
Retrons are bacterial immune systems that prevent the spread of phages by initiating a toxic response within infected hosts. All previously characterized retrons produce high levels of multicopy single-stranded DNA (msDNA) in the cell by reverse transcription, which acts as an antitoxin in the absence of phage infection. However, we describe here a non-canonical mechanism for Type VI retrons, which do not produce detectable msDNA in the absence of phage, yet still provide phage defense. Focusing primarily on Retron-Vpa2, a Type VI retron from Vibrio parahaemolyticus, we show broad defense against phages and identify triggers of the system within phage recombination systems. Within the Retron-Vpa2 operon, we find a highly enriched, structured transcript that we term a hybrid RNA (hyRNA), which contains both the retrons reverse transcription template and a translationally repressed toxic effector coding sequence. We find that phage infection induces the accumulation of high levels of msDNA and that this msDNA is necessary for derepressing translation of the antiviral toxin. These findings present key biological and mechanistic insights into a distinct group of retrons while highlighting the diversity of systems that participate in bacterial immunity.
]]></description>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Rojas-Montero, M.</dc:creator>
<dc:creator>Poola, D.</dc:creator>
<dc:creator>Klas, J.</dc:creator>
<dc:creator>Carabias, A.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Mestre, M. R.</dc:creator>
<dc:creator>Zhao, R.</dc:creator>
<dc:creator>Davila-Hidalgo, A.</dc:creator>
<dc:creator>Montoya, G.</dc:creator>
<dc:creator>Pinilla-Redondo, R.</dc:creator>
<dc:creator>Gonzalez-Delgado, A.</dc:creator>
<dc:creator>Shipman, S. L.</dc:creator>
<dc:date>2025-10-22</dc:date>
<dc:identifier>doi:10.1101/2025.10.22.683967</dc:identifier>
<dc:title><![CDATA[Reverse transcribed ssDNA derepresses translation of a retron antiviral protein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.22.684023v1?rss=1">
<title>
<![CDATA[
Germline-targeting HIV immunogen induces cross-neutralizing antibodies in outbred macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.22.684023v1?rss=1</link>
<description><![CDATA[
Germline-targeting-(GT) is a promising strategy to activate rare broadly neutralizing antibody (bnAb)-producing B cells against HIV, but induction of such responses in outbred animals has not been achieved. Using antibody-guided structure-based design, we engineered a germline-targeting trimer immunogen Q23-APEX-GT2 that primes diverse V2-apex bnAb precursors. Q23-APEX-GT2 efficiently activated V2-apex-specific B cells in humanized knock-in mice and consistently elicited immunofocused antibody responses in rhesus macaques, priming multiple long CDRH3-loop bnAb-B cell lineages. Monoclonal antibodies from immunized macaques exhibited broad heterologous HIV trimer binding and cross-neutralization. Atomic-level structural studies confirmed precise epitope targeting and revealed CDRH3-paratope configurations that mirrored those of human V2-apex bnAbs. This study provides proof-of-principle for successful priming and maturation of authentic V2-apex bnAb precursors in outbred macaques, underscoring the potential of V2-apex-targeted vaccines.

HIGHLIGHTSO_LIEngineered Q23-APEX-GT2 trimer to stimulate diverse V2-apex bnAb B cell precursors
C_LIO_LIQ23-APEX-GT2 primed rare V2-apex bnAb B cells in mice and outbred rhesus macaques
C_LIO_LIQ23-APEX-GT2 elicited immunofocused antibody responses and diverse V2-apex B cell lineages with desirable long-CDRH3 paratope properties
C_LIO_LIQ23-APEX-GT2 alone induced V2-apex antibodies with broad HIV trimer binding and modest neutralization breadth
C_LIO_LIStructural analysis confirmed bnAb site targeting, mirroring human and rhesus V2-apex bnAbs
C_LI
]]></description>
<dc:creator>Mishra, N.</dc:creator>
<dc:creator>Liang, B.</dc:creator>
<dc:creator>Roark, R. S.</dc:creator>
<dc:creator>Ghosh, A.</dc:creator>
<dc:creator>Callaghan, S.</dc:creator>
<dc:creator>Lee, W.-H.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Vo, A. L.</dc:creator>
<dc:creator>Avillion, G.</dc:creator>
<dc:creator>Habib, R. H.</dc:creator>
<dc:creator>Bibollet-Ruche, F.</dc:creator>
<dc:creator>Giese, G.</dc:creator>
<dc:creator>Somanathan, A.</dc:creator>
<dc:creator>Chowdhury, R. R.</dc:creator>
<dc:creator>Oberoi, P.</dc:creator>
<dc:creator>Amereh, K.</dc:creator>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Kassab, M.</dc:creator>
<dc:creator>Tjio, L.</dc:creator>
<dc:creator>Andrabi, S.</dc:creator>
<dc:creator>Reyes, R. A.</dc:creator>
<dc:creator>Allen, J. D.</dc:creator>
<dc:creator>James, N. E.</dc:creator>
<dc:creator>Randall, K. N.</dc:creator>
<dc:creator>van der Maas, L.</dc:creator>
<dc:creator>Ben-Akiva, E.</dc:creator>
<dc:creator>Kaczmarek-Michaels, K.</dc:creator>
<dc:creator>plante, S.</dc:creator>
<dc:creator>Martella, C. L.</dc:creator>
<dc:creator>Skelly, A. N.</dc:creator>
<dc:creator>Singh, A.</dc:creator>
<dc:creator>Hurtado, J.</dc:creator>
<dc:creator>Dueker, K.</dc:creator>
<dc:creator>Capozzola, T.</dc:creator>
<dc:creator>Nedellec, R.</dc:creator>
<dc:creator>Ozorowski, G.</dc:creator>
<dc:creator>Lewis, M.</dc:creator>
<dc:creator>Falcone, S.</dc:creator>
<dc:creator>Carfi, A.</dc:creator>
<dc:creator>Himansu, S.</dc:creator>
<dc:creator>Shapiro, L.</dc:creator>
<dc:creator>Crispin, M.</dc:creator>
<dc:creator>Hahn, B. H.</dc:creator>
<dc:creator>Briney,</dc:creator>
<dc:date>2025-10-23</dc:date>
<dc:identifier>doi:10.1101/2025.10.22.684023</dc:identifier>
<dc:title><![CDATA[Germline-targeting HIV immunogen induces cross-neutralizing antibodies in outbred macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.23.684104v1?rss=1">
<title>
<![CDATA[
Coupling differential adhesion to cell signaling avoids kinetic traps to yield robust multicellular self-organization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.23.684104v1?rss=1</link>
<description><![CDATA[
Differential adhesion, where cells physically reorganize based on their heterogeneous adhesion preferences, is one of the major models for self-organization in development and tissue formation. However, accumulating evidence suggests that differential adhesion is many times insufficient for robust convergence to a target minimal energy multicellular structure. Here we use computational simulations and engineered synthetic cell circuits to systematically explore alternative mechanisms for programming formation of a simple two-cell type core-shell morphology. Starting with two pre-differentiated cell types with constitutively high differential adhesion leads to kinetic trapping in variable, multi-core structures. In contrast, hybrid mechanisms that gradually induce differential adhesion upon cell-cell contact signaling consistently converge to the target single-core structure, in a manner robust to variation in cell numbers, interaction energy, and noise. This work delineates intrinsic limitations of self-organizing systems based solely on differential adhesion, and shows how inducible systems provide a way to invoke the strong adhesion required to maintain a multicellular structure, while avoiding the pitfall of kinetic traps. This study illustrates how joint computational and experimental exploration of synthetic circuits can be used to probe key developmental principles and tradeoffs and inform the design of synthetic development and self-organization.
]]></description>
<dc:creator>Pelc, G.</dc:creator>
<dc:creator>McKeithan, W. L.</dc:creator>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Brenner, M. P.</dc:creator>
<dc:creator>Lim, W. A.</dc:creator>
<dc:creator>Nitzan, M.</dc:creator>
<dc:date>2025-10-24</dc:date>
<dc:identifier>doi:10.1101/2025.10.23.684104</dc:identifier>
<dc:title><![CDATA[Coupling differential adhesion to cell signaling avoids kinetic traps to yield robust multicellular self-organization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.24.684275v1?rss=1">
<title>
<![CDATA[
STING activation reshapes the tumor microenvironment leading to tumor regression in osteosarcoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.24.684275v1?rss=1</link>
<description><![CDATA[
Osteosarcomas are characterized by a high degree of aneuploidy, chromothripsis and micronuclei, yet these tumors typically have an immunosuppressive, macrophage-rich, T-cell depleted tumor microenvironment. cGAS-STING dysregulation is a possible mechanism by which immune activation in response to tumor genomic instability could be repressed. We identified almost universal repression of cGAS or STING in human osteosarcomas. However, a STING-activation gene signature was predictive of survival in osteosarcoma patients suggesting potential for activation of this pathway in the osteosarcoma tumor microenvironment. Indeed, in immunocompetent osteosarcoma models, systemic STING agonism led to complete regression and induced lasting immunologic memory. Host STING activation is sufficient to promote this anti-tumor immunity even in the absence of tumor STING. These results nominate the cGAS-STING pathway as an important therapeutic target in osteosarcoma, a disease in which no new curative therapies have been developed in the last 40 years.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=138 SRC="FIGDIR/small/684275v1_ufig1.gif" ALT="Figure 1">
View larger version (38K):
org.highwire.dtl.DTLVardef@15b1e5corg.highwire.dtl.DTLVardef@12650aaorg.highwire.dtl.DTLVardef@79ef71org.highwire.dtl.DTLVardef@15704c9_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Young, E. P.</dc:creator>
<dc:creator>Johnson, C. A.</dc:creator>
<dc:creator>Lee, A. G.</dc:creator>
<dc:creator>Morris, D.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Breese, M. R.</dc:creator>
<dc:creator>Camara Serrano, J. A.</dc:creator>
<dc:creator>Schott, C. R.</dc:creator>
<dc:creator>Abram, C. L.</dc:creator>
<dc:creator>Leung, S. G.</dc:creator>
<dc:creator>Sayles, L. C.</dc:creator>
<dc:creator>Pettiwala, A. M.</dc:creator>
<dc:creator>Gutierrez Vera, V. S.</dc:creator>
<dc:creator>Lowell, C. A.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>McEachron, T. A.</dc:creator>
<dc:creator>Sweet-Cordero, E. A.</dc:creator>
<dc:date>2025-10-25</dc:date>
<dc:identifier>doi:10.1101/2025.10.24.684275</dc:identifier>
<dc:title><![CDATA[STING activation reshapes the tumor microenvironment leading to tumor regression in osteosarcoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.25.684215v1?rss=1">
<title>
<![CDATA[
Structure-guided design of native-like HIV Env Single Chain Trimers for enhanced stability, immunogenicity, and versatile vaccine delivery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.25.684215v1?rss=1</link>
<description><![CDATA[
Stabilizing the HIV envelope (Env) trimer in its native prefusion conformation is key to eliciting broadly neutralizing antibodies (bnAbs). We present a generalizable single-chain trimer-(SCT) design platform that enables the production of stable, native-like Env trimers across diverse HIV strains. Using a structure-guided approach, we developed CAP256.SU-SCT9, based on a CAP256.SU Env shown to induce V2-apex bnAbs in humans and rhesus macaques. CAP256.SU-SCT9 trimer exhibited native-like structure, bnAb-specific antigenicity, authentic glycosylation, and compatibility with nucleic acid and nanoparticle delivery. We extended this design strategy to a panel of HIV Envs representing global diversity, and immunogens known to initiate bnAb responses in humans. Structural analyses confirmed that SCTs consistently maintained prefusion-closed conformations, favorable glycan profiles, strong binding to trimer-specific bnAbs, and low reactivity with non-neutralizing antibodies. These findings establish SCT as a versatile platform for HIV trimer immunogen design, with potential to support next-generation vaccines aimed at eliciting protective bnAb responses.
]]></description>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Krishnamurthy, I.</dc:creator>
<dc:creator>Mishra, N.</dc:creator>
<dc:creator>James, N. E.</dc:creator>
<dc:creator>Roy Chowdhury, R.</dc:creator>
<dc:creator>Callaghan, S.</dc:creator>
<dc:creator>Somanathan, A.</dc:creator>
<dc:creator>Avillion, G.</dc:creator>
<dc:creator>Amereh, K.</dc:creator>
<dc:creator>Sekar, T.</dc:creator>
<dc:creator>Oberoi, P.</dc:creator>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Torres, J. L.</dc:creator>
<dc:creator>Lee, W.-H.</dc:creator>
<dc:creator>Allen, J. D.</dc:creator>
<dc:creator>Liang, B.</dc:creator>
<dc:creator>Giese, G.</dc:creator>
<dc:creator>Vo, A. L.</dc:creator>
<dc:creator>Andrabi, S.</dc:creator>
<dc:creator>Jain, I.</dc:creator>
<dc:creator>Mergiya, A.</dc:creator>
<dc:creator>Capozzola, T.</dc:creator>
<dc:creator>He, W.-t.</dc:creator>
<dc:creator>Anzanello, F.</dc:creator>
<dc:creator>Habib, R.</dc:creator>
<dc:creator>Bibollet-Ruche, F.</dc:creator>
<dc:creator>Nedellec, R.</dc:creator>
<dc:creator>Duttonsackett, S.</dc:creator>
<dc:creator>Ozorowski, G.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Hahn, B. H.</dc:creator>
<dc:creator>Shaw, G. M.</dc:creator>
<dc:creator>Ward, A. B.</dc:creator>
<dc:creator>Crispin, M.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Andrabi, R.</dc:creator>
<dc:date>2025-10-26</dc:date>
<dc:identifier>doi:10.1101/2025.10.25.684215</dc:identifier>
<dc:title><![CDATA[Structure-guided design of native-like HIV Env Single Chain Trimers for enhanced stability, immunogenicity, and versatile vaccine delivery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.26.684676v1?rss=1">
<title>
<![CDATA[
A20 restriction of nitric oxide production restores macrophage bioenergetic balance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.26.684676v1?rss=1</link>
<description><![CDATA[
Macrophages play critical roles in regulating host responses to microbial pathogens and other forms of tissue stress and can acquire pro-inflammatory or tissue reparative phenotypes. A20, or TNFAIP3, is a potent regulator of innate immune cell functions and is extensively linked to human inflammatory and autoimmune diseases. We now find that A20 is a powerful regulator of both glycolytic and mitochondrial respiration in macrophages. Differentiated macrophages that are acutely rendered A20 deficient exhibit increased glycolytic activity and markedly decreased mitochondrial respiration after LPS stimulation. These cells are unable to repolarize from an M1-like to M2-like phenotype. Compromised mitochondrial oxygen consumption in A20 deficient macrophages is caused by increased nitric oxide production. Inhibition or genetic ablation of inducible nitric oxide synthase (iNOS) restores mitochondrial oxidative phosphorylation and lactate production in these cells. These metabolic perturbations occur independently of exaggerated cytokine production and despite robust production of IL-10. Therefore, A20 prevents Warburg-like aerobic glycolysis and restores macrophage homeostasis.
]]></description>
<dc:creator>Ma, A.</dc:creator>
<dc:creator>Chan, M.-P.</dc:creator>
<dc:creator>Advincula, R.</dc:creator>
<dc:creator>Achacoso, P.</dc:creator>
<dc:creator>Grossman, E.</dc:creator>
<dc:creator>Nomura, D.</dc:creator>
<dc:creator>Malynn, B.</dc:creator>
<dc:date>2025-10-27</dc:date>
<dc:identifier>doi:10.1101/2025.10.26.684676</dc:identifier>
<dc:title><![CDATA[A20 restriction of nitric oxide production restores macrophage bioenergetic balance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.27.684845v1?rss=1">
<title>
<![CDATA[
Mutating different α-tubulin acetylation sites has distinct effects on axon terminal morphogenesis in Drosophila melanogaster 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.27.684845v1?rss=1</link>
<description><![CDATA[
Microtubules are created from uniform - and {beta}-tubulin building blocks but typically carry out a variety of specialized functions within a cell. The post-translational modification of tubulin is one means by which microtubule function can be tuned to match different cellular activities. While multiple sites of acetylation have been identified in tubulin, particularly -tubulin, the effect of acetylation at different sites on microtubule function remains poorly characterized. Here, we took a genetic approach in Drosophila to disrupt three conserved sites of acetylation (K326, K370, K401) in endogenous -tubulin and characterized the effects on neuronal development. Acetylation-blocking mutagenesis of -tubulin K326 (K326A) perturbed larval locomotion and reduced axon terminal growth at the neuromuscular junction. These deficits were accompanied by a reduction in stable microtubules, suggesting that the -tubulin K326A mutation exerts its effect by disrupting microtubule stability. In contrast, mutagenesis of -tubulin K370 and K401 had virtually no effect on microtubule stability, suggesting that the effects of these mutations on axon terminal morphogenesis and survival may be mediated through a different mechanism. Altogether, the varied effects of these mutations suggests that acetylation at these three different sites may regulate different aspects of microtubule function within developing neurons.
]]></description>
<dc:creator>Than, H.</dc:creator>
<dc:creator>Welch, C. J.</dc:creator>
<dc:creator>Schauer, E.</dc:creator>
<dc:creator>Trujillo, S. P.</dc:creator>
<dc:creator>Wildonger, J.</dc:creator>
<dc:date>2025-10-27</dc:date>
<dc:identifier>doi:10.1101/2025.10.27.684845</dc:identifier>
<dc:title><![CDATA[Mutating different α-tubulin acetylation sites has distinct effects on axon terminal morphogenesis in Drosophila melanogaster]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.27.684886v1?rss=1">
<title>
<![CDATA[
Development and validation of an ultra-low-cost, open source normothermic ex vivo organ perfusion platform 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.27.684886v1?rss=1</link>
<description><![CDATA[
BackgroundNormothermic ex vivo organ perfusion (NEVOP) promises to catalyze organ preservation, therapeutic discovery, and organ-specific disease modeling. Existing technology platforms remain inaccessible for research due to restricted access to commercial organ perfusion devices, high costs of both devices and proprietary consumables, and steep technical learning curves. Additionally, the available technology is not optimized to perfuse smaller organs such as the kidney.

MethodsTo overcome these barriers, a custom NEVOP circuit was developed using recycled, repurposed, and low-cost components. Porcine kidneys and autologous blood were used to iteratively optimize circuit design. A porcine kidney autotransplantation protocol was adapted to evaluate in vivo kidney function after ex vivo perfusion. To pilot the flexibility of this system as a multi-organ platform for ex vivo human biology, non-transplantable human donor kidney, spleen, and pancreas specimens were stably perfused using human blood products and analyzed.

ResultsAn ultra low-cost NEVOP system engineered to perfuse porcine kidneys and diverse human organs (kidney, pancreas, and spleen) sustained viable organs for up to 24 hours with evidence of both function and viability. Key innovations included a parallel flow resistor to facilitate low-flow perfusion in non-heparinized organs and a containment bag with adjustable magnets to provide vascular stability and recycling of venous overflow. The circuit costs less than 1,500USD to construct, and porcine kidneys perfused for 24 hours on this platform demonstrated healthy in vivo function upon autotransplantation.

ConclusionsCustom NEVOP platforms constitute novel and potentially transformative research platforms which use low-cost and readily available materials. Paired with access to non-transplantable research organs from altruistic donors, this model provides a road map for investigators to advance biomedical discovery and human ex vivo biology.
]]></description>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Higgins, N.</dc:creator>
<dc:creator>Chu, S.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Meyer, N. R.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>Saeed, M.</dc:creator>
<dc:creator>Ferreira, R.</dc:creator>
<dc:creator>Sorrentino, T. A.</dc:creator>
<dc:creator>Mena, J.</dc:creator>
<dc:creator>Suarez, P.</dc:creator>
<dc:creator>Maluf, F. C.</dc:creator>
<dc:creator>Sui, W.</dc:creator>
<dc:creator>Escobar, M. C.</dc:creator>
<dc:creator>Mann, U.</dc:creator>
<dc:creator>Braun, H.</dc:creator>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Elmer, J. R.</dc:creator>
<dc:creator>Chi, T. L.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:creator>Flake, A.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Stoller, M.</dc:creator>
<dc:date>2025-10-28</dc:date>
<dc:identifier>doi:10.1101/2025.10.27.684886</dc:identifier>
<dc:title><![CDATA[Development and validation of an ultra-low-cost, open source normothermic ex vivo organ perfusion platform]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.27.684958v1?rss=1">
<title>
<![CDATA[
Macrophage EHD1 promotes inflammation and stabilizes sortilin to accelerate atherosclerosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.27.684958v1?rss=1</link>
<description><![CDATA[
BackgroundMacrophages are key players in the pathogenesis of atherosclerosis. They trigger immune responses through their cell-surface receptors. However, how macrophages regulate those receptors in response to pro-inflammatory stimuli is not completely understood. Endocytic membrane trafficking involving receptor internalization, followed by endosomal transport and recycling of the internalized receptors, plays essential roles in balancing cell-surface receptors to meet cellular needs. Here, we explored the role of the endocytic regulator EHD1 in immune responses in macrophages and determined its contribution to atherosclerosis progression.

MethodsEHD1 expression profiles in mouse and human plaques were determined by single-cell RNA sequencing (scRNA-seq) and immunofluorescence staining. Bone marrow transplantation (BMT) by transplanting bone marrow cells from Ehd1-/- or littermate wild-type mice to irradiated Ldlr-/- mice was performed to determine the effect of EHD1 deletion on atherosclerosis progression. In vitro mechanistic studies including inflammation signaling and endocytosis assays were performed in bone marrow-derived macrophages.

ResultsEHD1 expression in macrophages is enhanced as atherosclerosis progresses in both mice and humans. Histological analysis of aortic root sections from BMT mice showed that EHD1 deletion reduces lesion size. ScRNA-seq of aortic CD45+ cells demonstrated that EHD1 deletion attenuates pro-inflammatory responses and cell-cell interactions. Mechanistic studies revealed that EHD1 accelerates the endocytic recycling of TNFR2 and activates NF-kB, leading to increased expression of inflammatory cytokines. Moreover, EHD1 interacts with retromer and stabilizes sortilin, a retrograde cargo of retromer and a risk factor for atherosclerosis.

ConclusionsEHD1 promotes inflammation by enhancing TNFR2-NF-kB signaling and stabilizing sortilin, leading to accelerated atherosclerosis. Our study reveals novel roles for EHD1-mediated membrane trafficking in macrophage function and paves the way to innovative therapeutic strategies that aim to address dysregulated membrane trafficking in atherosclerosis.
]]></description>
<dc:creator>Cai, B.</dc:creator>
<dc:creator>Ma, F.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Gupta, N.</dc:creator>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Xiao, L.</dc:creator>
<dc:creator>Traylor, J.</dc:creator>
<dc:creator>Rom, O.</dc:creator>
<dc:creator>Kovacic, J.</dc:creator>
<dc:creator>Fidler, T.</dc:creator>
<dc:creator>Yurdagul, A.</dc:creator>
<dc:creator>Orr, A. W.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:date>2025-10-28</dc:date>
<dc:identifier>doi:10.1101/2025.10.27.684958</dc:identifier>
<dc:title><![CDATA[Macrophage EHD1 promotes inflammation and stabilizes sortilin to accelerate atherosclerosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.28.684953v1?rss=1">
<title>
<![CDATA[
Repeated evolutionary turnover of vertebrate skeletal muscle myosins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.28.684953v1?rss=1</link>
<description><![CDATA[
Myosin heavy chain proteins are essential for muscle contraction and nearly every physiological function in animals, but their diversity and evolution outside mammals is largely unknown. We comprehensively model the evolutionary history of over 1100 heavy-chain myosins. We find that skeletal muscle myosins are located in a conserved tandem gene array in all vertebrate species, but repeated gene duplication-loss turnover has surprisingly led to an independently evolved set of core skeletal muscle myosins in each major vertebrate group. Despite these separate derivations of these myosin subfamilies, each major vertebrate group exhibits consistent tissue-specific patterns of subfamily expression and specialized myosin subfamily expression in extreme muscles. Our results show that muscle evolution across vertebrates is not based in conserved orthologous motor myosins, as might be expected for such a core structural protein family. Instead, we find that skeletal muscle myosins have evolved as a shifting cluster of genes that is constantly changing and diversifying to balance the need to maintain core physiology, while innovating new physiological possibilities.
]]></description>
<dc:creator>Harvey, C. M.</dc:creator>
<dc:creator>Schuppe, E. R.</dc:creator>
<dc:creator>Brainard, M. S.</dc:creator>
<dc:creator>Fuxjager, M. J.</dc:creator>
<dc:creator>Pease, J. B.</dc:creator>
<dc:date>2025-10-28</dc:date>
<dc:identifier>doi:10.1101/2025.10.28.684953</dc:identifier>
<dc:title><![CDATA[Repeated evolutionary turnover of vertebrate skeletal muscle myosins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.28.684989v1?rss=1">
<title>
<![CDATA[
Hippo signaling differentially regulates distal progenitor subpopulations and their transitional states to construct the mammalian lungs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.28.684989v1?rss=1</link>
<description><![CDATA[
Lung size control and cell type specification are key unresolved issues. In this study, we have engineered mosaic patterns of Hippo signaling to reveal the developmental potential of SOX9+ progenitors at the distal lung buds. Our results show that the distal SOX9+ subdomain is sufficient to direct lung outgrowth through bifurcation, providing a mechanism for lung size control. Through single-cell analyses, we identify transitional cell states and candidates for promoting cell fates. Moreover, genetic analysis reveals that Hippo signaling induces distinct cell fates at different SOX9+ subdomains to produce the conducting airways and the alveolar epithelium. These results provide the first extensive map of the developmental paths of lung cells. Some of the developmental paths of transitional cell states in mice correspond to those in human lungs. Together, these studies provide mechanistic insight into how Hippo signaling controls the sequential expansion and differentiation of SOX9+ progenitors to construct the mammalian lungs.
]]></description>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Basak, M.</dc:creator>
<dc:creator>Zaher, Y.</dc:creator>
<dc:creator>Yao, E.</dc:creator>
<dc:creator>Wang, S.-A.</dc:creator>
<dc:creator>Aung, T.</dc:creator>
<dc:creator>Chuang, P.-T.</dc:creator>
<dc:date>2025-10-28</dc:date>
<dc:identifier>doi:10.1101/2025.10.28.684989</dc:identifier>
<dc:title><![CDATA[Hippo signaling differentially regulates distal progenitor subpopulations and their transitional states to construct the mammalian lungs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.27.684881v1?rss=1">
<title>
<![CDATA[
A proteomic signature of vascular dysfunction linked to tauopathy and degeneration in the aging brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.27.684881v1?rss=1</link>
<description><![CDATA[
Small vessel disease (SVD) impacts healthy aging of organs across the body, yet its contributions to adverse brain aging remain poorly defined. Here we show thromboinflammation, a core feature of SVD, as a driver of adverse brain aging. We identify cerebrospinal fluid fibrinogen as a marker of brain thromboinflammation and screen neurovascular biosignatures mediating its impact on synaptic vulnerability along the full spectrum of brain aging from cognitively typical, amyloid-negative to cognitively impaired, amyloid-positive older adults. We identified 53 proteins mediating fibrinogens effects on synaptic markers in 1,655 donors from three independent cohorts. Single-cell transcriptomic mapping revealed mediator enrichment in neurovascular unit cells. Pathway analysis demonstrated dysregulation of angiogenesis, fibrosis, and immune signaling. Vascular and microglial-enriched biosignatures associated with compromised white matter integrity. These findings indicate thromboinflammation as an early, amyloid-independent pathway to neurodegeneration and tauopathy, establishing vascular health as fundamental to preserving brain healthspan.
]]></description>
<dc:creator>Radabaugh, H. L.</dc:creator>
<dc:creator>Keller, J. N.</dc:creator>
<dc:creator>Radtke, C. H.</dc:creator>
<dc:creator>Kunney, C.</dc:creator>
<dc:creator>Karvelas, N.</dc:creator>
<dc:creator>Chan, H. W.</dc:creator>
<dc:creator>Brathaban, N.</dc:creator>
<dc:creator>Meyer-Franke, A.</dc:creator>
<dc:creator>Casaletto, K.</dc:creator>
<dc:creator>Zetterberg, H.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Cruchaga, C.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Akassoglou, K.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Elahi, F. M.</dc:creator>
<dc:date>2025-10-29</dc:date>
<dc:identifier>doi:10.1101/2025.10.27.684881</dc:identifier>
<dc:title><![CDATA[A proteomic signature of vascular dysfunction linked to tauopathy and degeneration in the aging brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.27.684888v1?rss=1">
<title>
<![CDATA[
Pitavastatin counteracts venetoclax resistance mechanisms in acute myeloid leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.27.684888v1?rss=1</link>
<description><![CDATA[
The BCL2 inhibitor venetoclax has therapeutic activity in several hematological malignancies. In acute myeloid leukemia (AML), venetoclax combined with hypomethylating agents is the standard of care for patients unfit for intensive chemotherapy, but intrinsic and acquired resistance are common. Loss of p53 function is strongly associated with venetoclax resistance, and adding venetoclax to 5-azacitidine provides no overall survival benefit in TP53-mutant AML. Other frequent mechanisms of venetoclax resistance in AML include FLT3 mutations, MCL-1 upregulation, and altered mitochondrial metabolism. Unfortunately, it has been challenging to develop agents that target these mechanisms directly and combinatorially. Here we report that pitavastatin, an inhibitor of HMG-CoA-reductase, promotes apoptosis and overcomes several venetoclax resistance mechanisms in human AML cells. At clinically achievable concentrations, pitavastatin treatment has potent cytotoxic activity in cells with mutations in TP53 or FLT3. The apoptotic mechanism involves p53-independent PUMA upregulation and reduced MCL-1 expression. Pitavastatin also suppresses mitochondrial gene expression and oxidative metabolism. The pro-apoptotic actions of pitavastatin depend on depletion of geranylgeranyl pyrophosphate (GGPP) and can be recapitulated by inhibiting GGPP synthase or geranylgeranyltransferase-1 enzymes. These results provide a mechanistic rationale for adding pitavastatin to AML regimens to prevent or overcome venetoclax resistance.
]]></description>
<dc:creator>Fruman, D.</dc:creator>
<dc:creator>Buono, R.</dc:creator>
<dc:creator>Juarez, D.</dc:creator>
<dc:creator>Paul, M.</dc:creator>
<dc:creator>Skuli, S. J.</dc:creator>
<dc:creator>Wong, I. B.</dc:creator>
<dc:creator>Tarnekar, I.</dc:creator>
<dc:creator>Ying, Z.</dc:creator>
<dc:creator>Le, I.</dc:creator>
<dc:creator>Wertheim, G.</dc:creator>
<dc:creator>Bakayoko, A.</dc:creator>
<dc:creator>Kruidenier, M.</dc:creator>
<dc:creator>Sebti, S.</dc:creator>
<dc:creator>Konopleva, M.</dc:creator>
<dc:creator>Fleischman, A. G.</dc:creator>
<dc:creator>Jang, C.</dc:creator>
<dc:creator>Carroll, M.</dc:creator>
<dc:date>2025-10-29</dc:date>
<dc:identifier>doi:10.1101/2025.10.27.684888</dc:identifier>
<dc:title><![CDATA[Pitavastatin counteracts venetoclax resistance mechanisms in acute myeloid leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.28.685144v1?rss=1">
<title>
<![CDATA[
Acetylation of Axonal G3BP1 through ELP3 Accelerates Axon Regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.28.685144v1?rss=1</link>
<description><![CDATA[
Nerve injury triggers localized translation of axonal mRNAs to respond to injury and nerve regeneration. The core stress granule protein G3BP1 sequesters axonal mRNAs in granules before and after axotomy. G3BP1 granule disassembly can be regulated by post-translational modifications, including phosphorylation of S149 phosphorylation and acetylation of human K376 (mouse K374). Axonal G3BP1 undergoes phosphorylation after axotomy, but acetylation of G3BP1 in axons was unknown. Here we show that rodent G3BP1 undergoes K374 acetylation after axotomy is ELP3-dependent, which enhances axonal protein synthesis, accelerates nerve regeneration, and supports functional recovery. ELP3-depleted neurons exhibit reduced axon growth and increased axonal G3BP1 granules. The proximal axons degenerate rapidly despite maintaining soma connectivity, an effect prevented by expression of acetylmimetic G3BP1.Together, these findings identify G3BP1 acetylation via ELP3 as a critical regulator of both axonal regeneration and neuronal resilience, revealing a post-translational mechanism that links stress granule regulation to neuronal repair and protection.
]]></description>
<dc:creator>Dalla Costa, I.</dc:creator>
<dc:creator>Michenfelder, E.</dc:creator>
<dc:creator>Siciliano, S.</dc:creator>
<dc:creator>Tapita, A.</dc:creator>
<dc:creator>Buchanan, C. N.</dc:creator>
<dc:creator>Vaughn, L. S.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Oses-Prieto, J. A.</dc:creator>
<dc:creator>Ma, C.</dc:creator>
<dc:creator>Thames, E.</dc:creator>
<dc:creator>Samra, N.</dc:creator>
<dc:creator>Ben-Dor, S.</dc:creator>
<dc:creator>Haffner-Krausz, R.</dc:creator>
<dc:creator>McElveen, M.</dc:creator>
<dc:creator>Smith, T. P.</dc:creator>
<dc:creator>Nawar, N.</dc:creator>
<dc:creator>Manaswiyoungkul, P.</dc:creator>
<dc:creator>Gunning, P. T.</dc:creator>
<dc:creator>Fainzilber, M.</dc:creator>
<dc:creator>Burlingame, A.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Perlson, E.</dc:creator>
<dc:creator>Sahoo, P. K.</dc:creator>
<dc:creator>Twiss, J. L.</dc:creator>
<dc:date>2025-10-30</dc:date>
<dc:identifier>doi:10.1101/2025.10.28.685144</dc:identifier>
<dc:title><![CDATA[Acetylation of Axonal G3BP1 through ELP3 Accelerates Axon Regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.30.685469v1?rss=1">
<title>
<![CDATA[
Plasmodium falciparum exploits NUAK1 to establish infection in human erythrocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.30.685469v1?rss=1</link>
<description><![CDATA[
The malaria parasite Plasmodium falciparum continues to demonstrate growing drug resistance, raising the need for innovative treatments. Host-directed therapeutics are emerging as a promising approach for many infectious diseases, but knowledge of critical host factors for malaria is limited. P. falciparum is an obligate intracellular parasite of human erythrocytes, suggesting it has evolved to exploit specific host pathways to establish infection. Here, we report that the AMPK-related kinase NUAK1 is a critical host factor for P. falciparum in erythrocytes and has potential as a therapeutic target. We show that NUAK1 is present in human erythrocytes and undergoes increased phosphorylation in P. falciparum-infected cells. Two highly selective NUAK1 inhibitors, HTH-01-015 and WZ4003, inhibited P. falciparum growth throughout its asexual life cycle, including during erythrocyte invasion. Chemoproteomic profiling confirmed the inhibitors selectivity for human NUAK1. We further show that treatment with the inhibitors reduces phosphorylation of the well-characterized NUAK1 substrate MYPT1 in erythroid cells. Moreover, we find that genetic overexpression of NUAK1 in erythroid cells partially rescues both the signaling and invasion phenotypes elicited by the small molecule inhibitors. These results establish a critical role for the NUAK1 signaling pathway in P. falciparum-infected erythrocytes and highlight its potential as a vulnerable target for host-directed malaria control.
]]></description>
<dc:creator>Navarrete, D. J.</dc:creator>
<dc:creator>Kim, C. Y.</dc:creator>
<dc:creator>McLelland, K.</dc:creator>
<dc:creator>Gonzalez Ramirez, M.</dc:creator>
<dc:creator>Baro, B.</dc:creator>
<dc:creator>Salinas, N. D.</dc:creator>
<dc:creator>Tolia, N. H.</dc:creator>
<dc:creator>Doerig, C.</dc:creator>
<dc:creator>Ong, S.-E.</dc:creator>
<dc:creator>Golkowski, M. G.</dc:creator>
<dc:creator>Egan, E. S.</dc:creator>
<dc:date>2025-10-30</dc:date>
<dc:identifier>doi:10.1101/2025.10.30.685469</dc:identifier>
<dc:title><![CDATA[Plasmodium falciparum exploits NUAK1 to establish infection in human erythrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.30.685624v1?rss=1">
<title>
<![CDATA[
Engineered orthogonal translation systems from metagenomic libraries expand the genetic code 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.30.685624v1?rss=1</link>
<description><![CDATA[
Genetic code expansion with non-canonical amino acids (ncAAs) opens new opportunities for the function and design of proteins by broadening their chemical repertoire. Unfortunately, ncAA incorporation is limited both by a small collection of orthogonal aminoacyl-tRNA synthetases (aaRSs) and tRNAs and by low-throughput methods to discover them. Here, we report the discovery, characterization, and engineering of a UGA suppressing orthogonal translation system mined from metagenomic data. We developed an integrated computational and experimental pipeline to profile the orthogonality of >200 tRNAs, test >1,250 combinations of aaRS:tRNA pairs, and identify the AP1 TrpRS:tRNATrpUCA as an orthogonal pair that natively encodes tryptophan at the UGA codon. We demonstrate that the AP1 TrpRS:tRNATrpUCA is highly active in cell-free and cellular contexts. We then use Ochre, a genomically recoded Escherichia coli strain that lacks UAG and UGA codons, to engineer an AP1 TrpRS variant capable of 5-hydroxytryptophan incorporation at an open UGA codon. We anticipate that our strategy of integrating metagenomic bioprospecting with cell-free screening and cell-based engineering will accelerate the discovery and optimization of orthogonal translation systems for genetic code expansion.
]]></description>
<dc:creator>Seki, K.</dc:creator>
<dc:creator>Nguyen, M. T. A.</dc:creator>
<dc:creator>Penev, P. I.</dc:creator>
<dc:creator>Isaacs, F. J.</dc:creator>
<dc:creator>Banfield, J. F.</dc:creator>
<dc:creator>Jewett, M. C.</dc:creator>
<dc:date>2025-10-30</dc:date>
<dc:identifier>doi:10.1101/2025.10.30.685624</dc:identifier>
<dc:title><![CDATA[Engineered orthogonal translation systems from metagenomic libraries expand the genetic code]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.29.685380v1?rss=1">
<title>
<![CDATA[
PCSK9 and High-Fat Diet Synergistically Induce Neurovascular Dysfunction and Neuroinflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.29.685380v1?rss=1</link>
<description><![CDATA[
Cerebral small vessel disease (CSVD) is strongly linked to metabolic risk factors and represents a major cause of vascular cognitive impairment and dementia. The interactions of genetic and environmental risk factors driving cerebrovascular pathology in metabolic syndrome are poorly understood. Here, we characterize neuroinflammatory and neurodegenerative phenotypes in a mouse model of metabolic syndrome with atherosclerosis induced by hepatic proprotein convertase subtilisin/kexin type 9 (PCSK9) overexpression combined with high-fat diet (HFD). PCSK9+HFD mice exhibit hallmark features of CSVD including vascular rarefaction, impaired neurovascular coupling, blood-brain barrier disruption, white matter injury, neuronal loss, and cognitive deficits. Notably, we identify lipid-droplet accumulating microglia (LDAM) as a distinct cellular phenotype that emerges in response to metabolic stress and correlates with cerebrovascular dysfunction. Three-dimensional light sheet microscopy reveals widespread vascular network disruption. Immunophenotyping demonstrates that microglia in PCSK9+HFD group exhibit enhanced phagocytic activation and ramification complexity yet accumulate perivascular amyloid-{beta}, suggesting impaired clearance capacity. Importantly, we observed vascular amyloid-{beta} deposition in wild-type mice without genetic Alzheimers disease mutations, suggesting that metabolic stress contributes to cerebrovascular amyloid pathology. PCSK9+HFD mice displayed recognition memory deficits and increased anxiety-like behavior. Our findings establish that severe hypercholesterolemia accelerates CSVD pathogenesis, and identify LDAM as a distinct pathological feature linking systemic metabolic syndrome to cerebrovascular dysfunction and cognitive impairment.
]]></description>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Letian, A.</dc:creator>
<dc:creator>Mohl, G. A.</dc:creator>
<dc:creator>Kroke, B.</dc:creator>
<dc:creator>Arkelius, K.</dc:creator>
<dc:creator>Clelland, C. D.</dc:creator>
<dc:creator>Goldberg, E. L.</dc:creator>
<dc:creator>Singhal, N. S.</dc:creator>
<dc:date>2025-10-31</dc:date>
<dc:identifier>doi:10.1101/2025.10.29.685380</dc:identifier>
<dc:title><![CDATA[PCSK9 and High-Fat Diet Synergistically Induce Neurovascular Dysfunction and Neuroinflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.30.685539v1?rss=1">
<title>
<![CDATA[
Cohesin Acetylation and ATPase Activity Control Cohesion and Loop Architecture through Distinct Mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.30.685539v1?rss=1</link>
<description><![CDATA[
Cohesin is a conserved protein complex that mediates sister chromatid cohesion, chromosome condensation, gene regulation, and DNA repair. These processes rely on cohesins ability to tether sister chromatids and form chromatin loops, which depend on cohesins ATPase activity and Eco1-mediated acetylation of two lysine residues (K112 and K113 in budding yeast) in its Smc3 subunit. How cohesins ATPase activity and acetylation integrate to control cohesin functions remains poorly understood. Here, we analyzed chromatin architecture in yeast mutants with altered cohesin acetylation and/or ATPase activity. We find that acetylation of either K112 or K113 is sufficient to produce a wild-type chromosome structure with loops positioned at cohesin-associated regions (CARs), whereas loss of acetylation at both residues abolishes positioned loops, indicating that acetylation at either lysine alone can maintain wild-type chromatin architecture. We further show that a cohesin acetylation mutant, despite being defective in sister-chromatid tethering and thus failing to establish cohesion, still forms wild-type-like loops, while cohesion-competent mutants lack positioned loops. These results suggest that the activities required for cohesion and loop formation are mechanistically separable, arguing against passive loop capture. Moreover, a mutant with reduced ATPase activity showed a loop profile similar to wild type, indicating that cohesin with lower ATPase activity can still form wild-type chromatin architecture. By contrast, hyper-ATPase mutants accumulate positioned loops, suggesting that increasing ATPase activity can enhance loop processivity. Together, our findings support a multilayered regulatory model in which acetylation fine-tunes ATPase output and cohesin functions to shape genome architecture.
]]></description>
<dc:creator>Costantino, L.</dc:creator>
<dc:creator>Ye, T.</dc:creator>
<dc:creator>Boardman, K.</dc:creator>
<dc:creator>Xiang, S.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Mu, Y.</dc:creator>
<dc:creator>Ma, W.</dc:creator>
<dc:creator>Koshland, D.</dc:creator>
<dc:date>2025-10-31</dc:date>
<dc:identifier>doi:10.1101/2025.10.30.685539</dc:identifier>
<dc:title><![CDATA[Cohesin Acetylation and ATPase Activity Control Cohesion and Loop Architecture through Distinct Mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.31.685880v1?rss=1">
<title>
<![CDATA[
Single-cell, multi-region profiling of the macaque brain across the lifespan 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685880v1?rss=1</link>
<description><![CDATA[
Brain aging is a complex process with profound health and societal consequences. However, the molecular and cellular pathways that govern its temporal progression-along with any cell type-, region-, and sex-specific heterogeneity in such progression-remain poorly defined. Here, we present a transcriptomic atlas of 5.3 million cells from 582 samples spanning 11 brain regions of 55 rhesus macaques (29 female, 26 male), aged 5 months (early life) to 21 years (late adulthood). We annotate 12 major cell classes and 225 subclusters, including region-specific subtypes of excitatory and inhibitory neurons, astrocytes, and ependymal cells. We identify a vulnerable excitatory neuron population in the superficial cortical lamina and a cortical interneuron population that are less abundant later in life, along with subtle, region-specific, age-associated compositional differences in subpopulations of microglia and oligodendrocytes, whose detection required single-cell resolution. Finally, we chart convergent and divergent age-associated molecular signatures across brain regions and cell classes-where some of these signatures are sex-specific and could underlie sex biases in neurological disorders. We find that age-associated transcriptional programs not only overlap substantially with those seen in Alzheimers disease (AD), but also unfold along distinct temporal trajectories across brain regions, suggesting that aging and AD may share molecular roots that emerge at different life stages and in region-specific, sex-specific windows of vulnerability. This work provides a temporal, regional, and sex-stratified atlas of the aging primate brain, offering insights into cell type-specific vulnerabilities and regional heterogeneity with translational human relevance.
]]></description>
<dc:creator>Yang, W.</dc:creator>
<dc:creator>Watkins, K. L.</dc:creator>
<dc:creator>DeCasien, A. R.</dc:creator>
<dc:creator>O'Neill, M. B.</dc:creator>
<dc:creator>Bohlen, M. O.</dc:creator>
<dc:creator>O'Day, D. R.</dc:creator>
<dc:creator>Duran, M.</dc:creator>
<dc:creator>Qiu, C.</dc:creator>
<dc:creator>Meleshko, A.</dc:creator>
<dc:creator>Vo, A.</dc:creator>
<dc:creator>Menke, M.</dc:creator>
<dc:creator>Calderon, D.</dc:creator>
<dc:creator>Cayo Biobank Research Unit,</dc:creator>
<dc:creator>Sallet, J.</dc:creator>
<dc:creator>Higham, J. P.</dc:creator>
<dc:creator>Martinez, M. I.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Starita, L. M.</dc:creator>
<dc:creator>Montague, M. J.</dc:creator>
<dc:creator>Platt, M.</dc:creator>
<dc:creator>Chiou, K. L.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Snyder-Mackler, N.</dc:creator>
<dc:date>2025-11-01</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685880</dc:identifier>
<dc:title><![CDATA[Single-cell, multi-region profiling of the macaque brain across the lifespan]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.31.685930v1?rss=1">
<title>
<![CDATA[
Structural Mechanism of Prestin-Membrane Mechanotransduction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685930v1?rss=1</link>
<description><![CDATA[
Sound frequency discrimination in mammals depends on the conformational transitions of prestin (SLC26A5), the piezoelectric motor in outer hair cells. The mechanism that enables prestins electrically driven interconversion and its dependence on membrane mechanics, remains unresolved. Here, we show that membrane forces represent a strong driver of the same conformational changes generated by transmembrane voltage and stabilized by bound anions. Single particle cryo-EM structures of nanodisc-reconstituted prestin were obtained from varying lipid composition and membrane thickness. These structures show that membrane thinning biases prestin from a compact conformation to a fully expanded conformation, mimicking outer hair cell elongation/contraction during electromotility. In contrast, zebrafish SLC26A5 transporters undergo complete elevator movements with redistribution of areal changes across leaflets. The structures, together with mutagenesis, H/D exchange mass spectrometry data, and NLC measurements, offer a high-resolution understanding of how prestin translates membrane tension into charge and motor movement during sound-evoked vibrations, revealing a process of reciprocal electro-mechanical transduction essential for tuning cochlear amplification.
]]></description>
<dc:creator>Bavi, N.</dc:creator>
<dc:creator>Haller, P.</dc:creator>
<dc:creator>Homma, K.</dc:creator>
<dc:creator>Lin, X.</dc:creator>
<dc:creator>Milewski, W.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Sosnick, T.</dc:creator>
<dc:creator>Perozo, E.</dc:creator>
<dc:date>2025-11-01</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685930</dc:identifier>
<dc:title><![CDATA[Structural Mechanism of Prestin-Membrane Mechanotransduction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.31.685963v1?rss=1">
<title>
<![CDATA[
TOMM40 suppression promotes neuronal cholesterol imbalance and molecular and behavioral phenotypes of Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685963v1?rss=1</link>
<description><![CDATA[
INTRODUCTIONWhile the APOE4 allele is a major risk factor for Alzheimers disease (AD), the role of TOMM40--an adjacent gene involved in mitochondrial protein import--is not known.

METHODSMice, human iPSC-derived neurons (iNeurons), and human brain tissue were used for study of animal cognition, cholesterol metabolism, mitochondrial function, and gene expression.

RESULTSTOMM40 knockdown (KD) impaired memory in mice and increased cholesterol and A{beta} 42 in mouse brains and human iNeurons. KD disrupted mitochondria-endoplasmic reticulum contact sites (MERCs), causing mitochondrial dysfunction and promoting reactive oxygen species that led to activation of LXRB (NR1H2), upregulation of APOE and LDLR. and increased cellular cholesterol and A{beta} 42 independent of APOE4. Human brain transcriptomics showed reduced TOMM40 expression that correlated with cholesterol regulatory gene expression, amyloid burden, and clinical AD diagnosis.

DISCUSSIONTOMM40 is a novel mediator of AD pathology through dual effects on MERCs that regulate cholesterol homeostasis and mitochondrial function.

GRAPHICAL ABSTRACT

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]]></description>
<dc:creator>Yang, N. V.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Simms, J.</dc:creator>
<dc:creator>Dinh, L.</dc:creator>
<dc:creator>Huang, A.</dc:creator>
<dc:creator>Oei, J. H.</dc:creator>
<dc:creator>Yassine, H. N.</dc:creator>
<dc:creator>Krauss, R. M.</dc:creator>
<dc:date>2025-11-01</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685963</dc:identifier>
<dc:title><![CDATA[TOMM40 suppression promotes neuronal cholesterol imbalance and molecular and behavioral phenotypes of Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.31.685897v1?rss=1">
<title>
<![CDATA[
Rapid generation of ventral A9-like dopaminergic neurons from patterned iPSCs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685897v1?rss=1</link>
<description><![CDATA[
In vitro modelling of highly vulnerable nigral dopaminergic (DA) neuronal subtypes in Parkinsons disease (PD), is necessary for studying disease mechanisms. Here, we optimized a new approach by expressing the pioneer neurogenic transcription factor, Achaete-scute-like 1 (Ascl1), implicated in determining dopaminergic fate. Sequential small-molecule patterning of iPSCs into early floor plate mesencephalic progenitors, followed by inducible Ascl1 expression, rapidly differentiates midbrain DA neurons. Immunocytochemistry and transcriptomic analysis of these patterned Ascl1-driven DA neurons (PA-DANs) confirmed midbrain-lineage specificity. Importantly, we found an enrichment of DA subpopulations that corresponded to the adult human ventral SOX6-positive A9 DA subtypes vulnerable in PD. Furthermore, we combined these ventral A9-like PA-DANs with human iPSC-derived midbrain astrocytes and microglia in defined ratios to generate mature 3D A9-like assembled organoids that display characteristic spontaneous neuronal activity and electrical propagation along the axon. Our method efficiently generates a mature and functional A9-like DA neuronal platform to study PD.

HighlightsO_LISequential midbrain patterning and Ascl1 expression accelerates DA differentiation
C_LIO_LIPA-DANs resemble human adult ventral A9-like DA subtypes vulnerable in PD
C_LIO_LI3D assembled organoids show mature identity of PA-DANs, iAstrocytes and iMicroglia
C_LIO_LIPA-DANs matured in 3D organoids show neuronal network activity within weeks
C_LI

eTOC blurbIn this study, Ullian and colleagues have developed a rapid method to differentiate dopaminergic neurons, using small molecules to generate early floor plate mesencephalic progenitors from human iPSCs and sequentially expressing a pioneer transcription factor, Ascl1, that accelerates uniform dopaminergic neurogenesis. Patterned Ascl1-driven dopaminergic neurons (PA-DANs) in 2D and 3D assembled organoids serve as a platform to study Parkinsons disease



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]]></description>
<dc:creator>Chaplot, K.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Wessman, J.</dc:creator>
<dc:creator>Rivera, M.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Duan, X.</dc:creator>
<dc:creator>England, P. M.</dc:creator>
<dc:creator>Clark, I. C.</dc:creator>
<dc:creator>Ullian, E. M.</dc:creator>
<dc:date>2025-11-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685897</dc:identifier>
<dc:title><![CDATA[Rapid generation of ventral A9-like dopaminergic neurons from patterned iPSCs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.31.685945v1?rss=1">
<title>
<![CDATA[
Spatial Attention and Session Day Independently Modulate Human Visual Cortical Plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685945v1?rss=1</link>
<description><![CDATA[
Stimulus-specific response potentiation (SSRP) is a noninvasive form of cortical neuroplasticity elicited by repeated presentation of high-contrast visual stimuli. Analogous to long-term potentiation, SSRP has been proposed to exhibit input specificity, with potentiation confined to neural populations driven by the induction stimulus. In rodent models, SSRP effects accumulate across days; however, it remains unclear whether selective attention influences the magnitude or specificity of potentiation in humans. Here, we examined whether covert spatial attention modulates SSRP strength using high-density electroencephalography (EEG) in neurotypical adults. An established SSRP paradigm was modified to include an attention task during induction. Pre- and post-induction amplitudes were measured using frequency-tagged (6 and 7.5 reversals/s) bilateral hemifield contrast-sweep checkerboard steady-state visual evoked potentials (SSVEPs). The 10-minute, high-contrast, 2 Hz sign-reversing induction stimulus was presented exclusively to the left hemifield, with the right serving as control. Across two experimental sessions, participants attended either toward or away from the potentiated hemifield during induction. SSRP produced increased post-induction amplitudes in both hemifields, challenging the notion of strict input specificity. Potentiation was significantly greater on the second session day, independent of attention condition. Notably, attention enhanced SSRP in naive but not experienced observers, reflecting a significant interaction between attention and session day. Together, these findings suggest that (1) human SSRP may not be strictly stimulus-specific, (2) attention modestly enhances SSRP during initial exposure, and (3) repeated induction produces a robust plasticity effect that occludes attentional modulation.
]]></description>
<dc:creator>Limon, P. N.</dc:creator>
<dc:creator>Norcia, A. M.</dc:creator>
<dc:creator>Ash, R. T.</dc:creator>
<dc:date>2025-11-03</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685945</dc:identifier>
<dc:title><![CDATA[Spatial Attention and Session Day Independently Modulate Human Visual Cortical Plasticity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.03.685577v1?rss=1">
<title>
<![CDATA[
Epigenetic control of microglial developmental milestones from proliferative progenitors to efficient phagocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.03.685577v1?rss=1</link>
<description><![CDATA[
Early immune perturbations increase the risk of neurodegenerative and neurodevelopmental disorders, yet the mechanisms underlying the maturation of microglia, the resident immune cells of the brain parenchyma, remain poorly defined. Specifically, how proliferation, morphological differentiation, and phagocytosis are coordinated among microglia progenitors as they colonize the embryonic brain remains unclear. Here, we combined mathematical modeling with spatiotemporal analyses of the murine hippocampus and cerebellum from postnatal day 2 (P2) to P60 to reconstruct the trajectory of microglial development. We identified a proliferative-to-quiescent (P/Q) switch around P3/P4 that preceded the acquisition of morphological complexity and efficient phagocytosis and was accompanied by coordinated shifts in cell-cycle dynamics and metabolic state. Strikingly, this P/Q switch was recapitulated in repopulation contexts in mice and in the human fetal brain, where later stages displayed enhanced phagocytic function coupled to reduced proliferation. Perturbing the proliferative phase through pharmacological or genetic disruption of CSF1R signaling impaired subsequent microglial complexity and phagocytosis efficiency, revealing an unexpected reliance of phagocytosis on proliferation-driven colonization. Finally, we show that microglia stepwise maturation during development is associated with chromatin remodeling and driven by the epigenetic regulator Ikaros. Together, these findings uncover the sequential milestones of microglial development, revealing a potential period of early vulnerability and establishing an unexpected linkage between proliferation and phagocytosis essential to understanding how these processes are coordinated in neurodegenerative disorders.
]]></description>
<dc:creator>Pereira-Iglesias, M.</dc:creator>
<dc:creator>Martinson, D.</dc:creator>
<dc:creator>Falco, C.</dc:creator>
<dc:creator>Maldonado-Teixido, J.</dc:creator>
<dc:creator>Gonzalez-Rodriguez, M.</dc:creator>
<dc:creator>Senovilla-Ganzo, R.</dc:creator>
<dc:creator>Benito, E.</dc:creator>
<dc:creator>Beccari, S.</dc:creator>
<dc:creator>Valero, J.</dc:creator>
<dc:creator>Mora-Romero, B.</dc:creator>
<dc:creator>Ballasch, I.</dc:creator>
<dc:creator>Viguier, S.</dc:creator>
<dc:creator>Haene, P.</dc:creator>
<dc:creator>Boettiger, M.</dc:creator>
<dc:creator>Reisz, J. A.</dc:creator>
<dc:creator>Manso, Y.</dc:creator>
<dc:creator>Parkkinen, L.</dc:creator>
<dc:creator>Aransay, A. M.</dc:creator>
<dc:creator>Soria, F. N.</dc:creator>
<dc:creator>D'Alessandro, A.</dc:creator>
<dc:creator>Soriano, E.</dc:creator>
<dc:creator>Thion, M.</dc:creator>
<dc:creator>Garel, S.</dc:creator>
<dc:creator>Greter, M.</dc:creator>
<dc:creator>Giralt, A.</dc:creator>
<dc:creator>Pascual, A.</dc:creator>
<dc:creator>Garcia-Moreno, F.</dc:creator>
<dc:creator>Menassa, D. A.</dc:creator>
<dc:creator>Carrillo, J. A.</dc:creator>
<dc:creator>Sierra, A.</dc:creator>
<dc:date>2025-11-04</dc:date>
<dc:identifier>doi:10.1101/2025.11.03.685577</dc:identifier>
<dc:title><![CDATA[Epigenetic control of microglial developmental milestones from proliferative progenitors to efficient phagocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.04.685750v1?rss=1">
<title>
<![CDATA[
NINJ1 is activated by Mycobacterium tuberculosis ESX-1 secreted effector EsxA and mediates necrosis of infected human macrophages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.04.685750v1?rss=1</link>
<description><![CDATA[
Lytic cell death is recognized as a driver of tissue damage and dissemination during Mycobacterium tuberculosis (Mtb) infection, yet its execution mechanisms remain incompletely defined. Here, we identify Ninjurin-1 (NINJ1), a mediator of plasma membrane rupture (PMR), as a central effector of macrophage lysis in response to Mtb. Infection triggered NINJ1 oligomerization, and genetic or pharmacological inhibition of NINJ1 markedly reduced lactate dehydrogenase (LDH) release. Surprisingly, individual or combined inhibition of pyroptosis, apoptosis, necroptosis, or ferroptosis failed to prevent PMR in Mtb-infected macrophages. Instead, the ESX-1-secreted effector EsxA directly induced NINJ1 oligomerization, implicating pathogen-driven membrane damage as the upstream trigger. NINJ1 activation was independent of calcium or high-MW PEG, although PEG reduced LDH-release, implicating swelling-induced membrane changes in PMR. Cytokine release was mostly NINJ1-independent, except for CXCL-10, which was diminished in NINJ1-deficient macrophages. These findings establish NINJ1 as a key executioner of lytic cell death during Mtb-infection.
]]></description>
<dc:creator>Saetra, R. S. R.</dc:creator>
<dc:creator>Hansen, M.</dc:creator>
<dc:creator>Kappelhoff, S.</dc:creator>
<dc:creator>Bugge, M.</dc:creator>
<dc:creator>Marstad, A.</dc:creator>
<dc:creator>Ryan, L.</dc:creator>
<dc:creator>Devant, P.</dc:creator>
<dc:creator>Kagan, J. C.</dc:creator>
<dc:creator>Freude, K. K.</dc:creator>
<dc:creator>Beckwith, K. S.</dc:creator>
<dc:creator>FLO, T. H.</dc:creator>
<dc:date>2025-11-04</dc:date>
<dc:identifier>doi:10.1101/2025.11.04.685750</dc:identifier>
<dc:title><![CDATA[NINJ1 is activated by Mycobacterium tuberculosis ESX-1 secreted effector EsxA and mediates necrosis of infected human macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.04.686591v1?rss=1">
<title>
<![CDATA[
A role for the Stentor syntaxin protein in post-wound cell survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.04.686591v1?rss=1</link>
<description><![CDATA[
Wound healing is an essential biological process that occurs both in tissues and single cells. In free-living single-celled ciliates such as Stentor coeruleus, rapid wound healing is necessary to repair breaches to the plasma membrane, where any delays represent the difference between life and death. In order to discover novel molecular pathways that are important for healing in Stentor, we carried out a targeted RNA interference-based perturbation genetic screen combined with microsurgical wounding using a microfluidic guillotine to introduce reproducible bisection wounds. We identified a Stentor syntaxin gene that was necessary for cell survival, particularly post-wounding, with only [~]37% of the syntaxin-deficient cells surviving compared with [~]98% of the control cells. Syntaxin-deficient cells were more susceptible to hyposmotic shock and became increasingly vacuolated in the hours post-wounding, eventually leading to cell death. Osmotic stabilization of the cells during and after bisection partially restored the post-wound survival in knockdown cells. These results support the interpretation that syntaxin-deficient cells lack essential membrane fusion machinery, which manifests in vacuolar defects, and are deficient in maintaining osmotic homeostasis necessary for their survival post-wounding. This study provides a template for the discovery of new wound healing biology in emerging model systems.

SignificanceO_LIStentor is a single-celled ciliate with prodigious healing ability, yet the molecular mechanisms by which the cell rapidly heals wounds are not fully characterized.
C_LIO_LIThrough a targeted microfluidics genetic screen, we identified a novel syntaxin-like protein that is essential for Stentor survival post-wounding.
C_LIO_LISyntaxin-deficient cells were more susceptible to osmotic shock. Wounding cells in osmotically balanced conditions reduced the degree of hyposmotic stress associated with wounding and enabled more cells to survive. Our discovery highlights the rebalancing of intracellular osmolarity as an important step in wound repair in freshwater-dwelling organisms.
C_LI
]]></description>
<dc:creator>Nadkarni, A. V.</dc:creator>
<dc:creator>Diaz, U.</dc:creator>
<dc:creator>Zhang, K. F.</dc:creator>
<dc:creator>Tang, S. K. Y.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2025-11-05</dc:date>
<dc:identifier>doi:10.1101/2025.11.04.686591</dc:identifier>
<dc:title><![CDATA[A role for the Stentor syntaxin protein in post-wound cell survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.04.686611v1?rss=1">
<title>
<![CDATA[
Large-scale discovery of neural enhancers for cis-regulation therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.04.686611v1?rss=1</link>
<description><![CDATA[
CRISPR-based gene activation (CRISPRa) has emerged as a promising therapeutic approach for neurodevelopmental disorders (NDD) caused by haploinsufficiency. However, scaling this cis-regulatory therapy (CRT) paradigm requires pinpointing which candidate cis-regulatory elements (cCREs) are active in human neurons, and which can be targeted with CRISPRa to yield specific and therapeutic levels of target gene upregulation. Here, we combine Massively Parallel Reporter Assays (MPRAs) and a multiplex single cell CRISPRa screen to discover functional human neural enhancers whose CRISPRa targeting yields specific upregulation of NDD risk genes. First, we tested 5,425 candidate neuronal enhancers with MPRA, identifying 2,422 that are active in human neurons. Selected cCREs also displayed specific, autonomous in vivo activity in the developing mouse central nervous system. Next, we applied multiplex single-cell CRISPRa screening with 15,643 gRNAs to test all MPRA-prioritized cCREs and 761 promoters of NDD genes in their endogenous genomic contexts. We identified hundreds of promoter- and enhancer-targeting CRISPRa gRNAs that upregulated 200 of the 337 NDD genes in human neurons, including 91 novel enhancer-gene pairs. Finally, we confirmed that several of the CRISPRa gRNAs identified here demonstrated selective and therapeutically relevant upregulation of SCN2A, CHD8, CTNND2 and TCF4 when delivered virally to patient cell lines, human cerebral organoids, and a humanized mouse model of hTcf4. Our results provide a comprehensive resource of active, target-linked human neural enhancers for NDD genes and corresponding gRNA reagents for CRT development. More broadly, this work advances understanding of neural gene regulation and establishes a generalizable strategy for discovering CRT gRNA candidates across cell types and haploinsufficient disorders.
]]></description>
<dc:creator>McDiarmid, T. A.</dc:creator>
<dc:creator>Page, N. F.</dc:creator>
<dc:creator>Chardon, F. M.</dc:creator>
<dc:creator>Daza, R. M.</dc:creator>
<dc:creator>Chen, G. T.</dc:creator>
<dc:creator>Kosicki, M.</dc:creator>
<dc:creator>James, L. M.</dc:creator>
<dc:creator>Nourie, H. C.</dc:creator>
<dc:creator>Laboy-Cintron, D.</dc:creator>
<dc:creator>Lee, A. S.</dc:creator>
<dc:creator>Vij, P.</dc:creator>
<dc:creator>Calderon, D.</dc:creator>
<dc:creator>Lalanne, J.-B.</dc:creator>
<dc:creator>Martin, B. K.</dc:creator>
<dc:creator>Fink, K.</dc:creator>
<dc:creator>Talkowski, M. E.</dc:creator>
<dc:creator>Muotri, A. R.</dc:creator>
<dc:creator>Philpot, B. D.</dc:creator>
<dc:creator>Pennacchio, L. A.</dc:creator>
<dc:creator>Geschwind, D. H.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:date>2025-11-05</dc:date>
<dc:identifier>doi:10.1101/2025.11.04.686611</dc:identifier>
<dc:title><![CDATA[Large-scale discovery of neural enhancers for cis-regulation therapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.04.686676v1?rss=1">
<title>
<![CDATA[
RainBar: Optical Barcoding for Pooled Live-Cell Imaging with Single-Cell Resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.04.686676v1?rss=1</link>
<description><![CDATA[
High-throughput pooled screening has advanced functional genomics, but most existing methods rely on endpoint sequencing and are blind to dynamic, time-resolved phenotypes. We developed RainBar (Rainbow Barcodes), an optical barcoding system that supports pooled live-cell imaging with single-cell resolution. RainBar uses lentiviral co-delivery of spectrally distinct nuclear and cytoplasmic fluorescent proteins to encode up to 64 unique perturbations per well. To mitigate barcode recombination and improve decoding accuracy, we employed single-template viral production, codon diversification, and a ratio-based spectral unmixing pipeline tailored to overlapping fluorophores. An inverted cytoplasmic segmentation approach and multilayer perceptron classifier enabled accurate barcode identification in both arrayed and pooled formats. As a proof of concept, we applied RainBar to dissect NF-{kappa}B signaling dynamics in epithelial cells. Live imaging of RelA translocation uncovered stimulus-specific kinetics: IL-1{beta} triggered rapid recovery, while TNF induced prolonged nuclear localization. In pooled CRISPRi screens, RainBar recovered known NF-{kappa}B regulators (e.g., IL1R1, MYD88, TNFRSF1A) and highlighted additional modulators, including the Ino80 chromatin remodeling complex subunits and KAT2A acetyltransferase. Together, these results position RainBar as a flexible platform for multiplexed, image-based functional genomics, with potential to reveal dynamic signaling architectures across diverse cellular contexts in live cells.
]]></description>
<dc:creator>Mosadeghi, R.</dc:creator>
<dc:creator>Foyt, D.</dc:creator>
<dc:creator>Sharp, L.</dc:creator>
<dc:creator>Taylor, C. A.</dc:creator>
<dc:creator>Tay, N.</dc:creator>
<dc:creator>Oberlin, S.</dc:creator>
<dc:creator>Fan, J.</dc:creator>
<dc:creator>Bourke, S.</dc:creator>
<dc:creator>Kattah, M. G.</dc:creator>
<dc:creator>Huang, B. Q.</dc:creator>
<dc:creator>McManus, M. T.</dc:creator>
<dc:date>2025-11-05</dc:date>
<dc:identifier>doi:10.1101/2025.11.04.686676</dc:identifier>
<dc:title><![CDATA[RainBar: Optical Barcoding for Pooled Live-Cell Imaging with Single-Cell Resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.05.685843v1?rss=1">
<title>
<![CDATA[
A spatial transcriptomic atlas of autism-associated genes identifies convergence in the developing human thalamus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.05.685843v1?rss=1</link>
<description><![CDATA[
Autism is a highly heritable neurodevelopmental condition that manifests across a wide phenotypic spectrum. Rare and de novo loss-of-function mutations strongly predispose to autism and co-occurring developmental and intellectual disabilities in over 10% of autistic individuals. Understanding whether these variants converge on specific regional brain circuits or widely alter human brain development is crucial to understanding the etiology of profound autism. To date, transcriptomic atlases have mainly implicated the developing cerebral cortex, yet other brain areas have received relatively little attention. Here, we present a single-cell resolution spatial transcriptomic atlas of 250 autism susceptibility genes during human brain development. Profiling over 10 million cells across the midgestation forebrain, we found convergence of these genes across a small number of regional programs. The developing thalamus showed the most prevalent expression of autism susceptibility genes, followed by germinal zones throughout the brain. Within the thalamus, excitatory neurons showed the most enriched expression, which varied across thalamic nuclei harboring distinct circuits. Across the germinal zones, neural progenitors in the medial ganglionic eminences that generate parvalbumin- and somatostanin-positive interneurons showed highest expression. Our findings reveal the prevalent expression of autism associated genes beyond the developing cerebral cortex and implicate the developing human thalamus as a major hub of autism susceptibility.
]]></description>
<dc:creator>Aivazidis, A.</dc:creator>
<dc:creator>Memi, F.</dc:creator>
<dc:creator>Rademaker, K.</dc:creator>
<dc:creator>Koko, M.</dc:creator>
<dc:creator>Roberts, K.</dc:creator>
<dc:creator>Trinh, A.</dc:creator>
<dc:creator>Petryszak, R.</dc:creator>
<dc:creator>Kleshchevnikov, V.</dc:creator>
<dc:creator>Tuck, E.</dc:creator>
<dc:creator>Lisgo, S.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Makarchuk, S.</dc:creator>
<dc:creator>Prete, M.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:creator>Martin, H. C.</dc:creator>
<dc:creator>Bayraktar, O. A.</dc:creator>
<dc:date>2025-11-06</dc:date>
<dc:identifier>doi:10.1101/2025.11.05.685843</dc:identifier>
<dc:title><![CDATA[A spatial transcriptomic atlas of autism-associated genes identifies convergence in the developing human thalamus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.05.684493v1?rss=1">
<title>
<![CDATA[
An InDel Genomic Variant within a Bifunctional Super-Enhancer for LINC00636 and CD47 Regulation in Breast Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.05.684493v1?rss=1</link>
<description><![CDATA[
Highly accessible genomic super-enhancers often drive tumor-promoting programs, yet the impact of genomic variation within these regulatory elements remains unclear. Here, we identified a bifunctional super-enhancer that regulates the expression of cancer-promoting genes LINC00636 and CD47 in breast cancer. We discovered that a common germline insertion variant within the super-enhancer is associated with reduced chromatin accessibility at the super-enhancer locus. Deletion of the insertion in breast cancer cells increased chromatin accessibility, leading to upregulation of LINC00636 and CD47, enhanced resistance to nutrient-deprivation-induced apoptosis (mediated by CD47), activation of senescence (driven by elevated LINC00636), delayed cell death, and reduced infiltration of CD80 pro-inflammatory macrophages, changes that represent tumor-promoting features. Together, our findings uncover a common insertion-deletion variant that fine-tunes the regulatory activity of a bifunctional super-enhancer, suggest a protective role for the insertion allele, and establish a novel function for LINC00636 in senescence and breast cancer.
]]></description>
<dc:creator>Di Benedetto, C.</dc:creator>
<dc:creator>Tsark, A.</dc:creator>
<dc:creator>Acenas, D. D.</dc:creator>
<dc:creator>Thach, A.</dc:creator>
<dc:creator>Singal, A.</dc:creator>
<dc:creator>Opazo-Medallo, V.</dc:creator>
<dc:creator>Rodriguez, A.</dc:creator>
<dc:creator>El Zeini, M.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Park, C.</dc:creator>
<dc:creator>Gonzales-Velozo, H.</dc:creator>
<dc:creator>Weissman, I. L.</dc:creator>
<dc:creator>Betancur, P.</dc:creator>
<dc:date>2025-11-07</dc:date>
<dc:identifier>doi:10.1101/2025.11.05.684493</dc:identifier>
<dc:title><![CDATA[An InDel Genomic Variant within a Bifunctional Super-Enhancer for LINC00636 and CD47 Regulation in Breast Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.05.686845v1?rss=1">
<title>
<![CDATA[
Disruption of Cell-Type-Specific Molecular Programs of Medium Spiny Neurons in Autism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.05.686845v1?rss=1</link>
<description><![CDATA[
Autism spectrum disorders (ASD) are highly heritable neurodevelopmental conditions with major contributions from rare genetic variants. Most studies have focused on cortical mechanisms; even growing evidence implicates subcortical circuits in ASD etiology. To systematically map developmental and molecular alterations beyond the cortex, we profiled lineage relationships across five brain regions in an ASD mouse model. Most prominent changes emerged in the striatum, a hub for learning and motor control. Furthermore, we performed single-nucleus multiomic profiling of human putamen from ASD and neurotypical donors revealed cell-type-specific transcriptomic and regulatory alterations. Differential expression converged on synaptic and energy metabolic dysfunctions in D1 striosome medium spiny neurons (MSNs), coupled with astrocytic remodeling of synaptic support. Gene regulatory network analysis identified EGR3 and EGR1 as key transcriptional regulators of ASD-associated programs of D1 MSNs. Together, these results establish the striatum as a central node of ASD convergence and provide a multiomic resource for dissecting its subcortical mechanisms.
]]></description>
<dc:creator>Yuan, G.</dc:creator>
<dc:creator>Suresh, V.</dc:creator>
<dc:creator>Wigdor, E.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Leonard, R.</dc:creator>
<dc:creator>Steyert, M.</dc:creator>
<dc:creator>Griffiths, M.</dc:creator>
<dc:creator>Evans, C.</dc:creator>
<dc:creator>Rohani, N.</dc:creator>
<dc:creator>Weiss, J.</dc:creator>
<dc:creator>Lassen, F. H.</dc:creator>
<dc:creator>Schafer, N.</dc:creator>
<dc:creator>Dong, S.</dc:creator>
<dc:creator>Palmer, D. S.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:date>2025-11-07</dc:date>
<dc:identifier>doi:10.1101/2025.11.05.686845</dc:identifier>
<dc:title><![CDATA[Disruption of Cell-Type-Specific Molecular Programs of Medium Spiny Neurons in Autism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.06.687082v1?rss=1">
<title>
<![CDATA[
SPARK: in silico simulations for benchmarking nascent RNA sequencing experiments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.06.687082v1?rss=1</link>
<description><![CDATA[
Nascent RNA sequencing offers profound insights into transcriptional dynamics, yet there are substantial challenges to analyzing these data. The development of proper computational tools necessitates realistic benchmarking datasets that reflect biological variability and technical biases. We present simulated pre-mRNA and RNA kinetics (SPARK), a versatile in silico framework for generating reads across nascent RNA sequencing approaches. SPARK simulates the process of transcription -- allowing for variable elongation rates and pausing events -- and key experimental features. SPARK provides a comprehensive platform for computational development and benchmarking in nascent RNA genomics.
]]></description>
<dc:creator>Calvo-Roitberg, E.</dc:creator>
<dc:creator>Lehman, J. W.</dc:creator>
<dc:creator>Tam, E.</dc:creator>
<dc:creator>Elhajjajy, S.</dc:creator>
<dc:creator>Engelhardt, B. E.</dc:creator>
<dc:creator>Pai, A. A.</dc:creator>
<dc:date>2025-11-08</dc:date>
<dc:identifier>doi:10.1101/2025.11.06.687082</dc:identifier>
<dc:title><![CDATA[SPARK: in silico simulations for benchmarking nascent RNA sequencing experiments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.08.687348v1?rss=1">
<title>
<![CDATA[
Structural basis and physiological significance of non-canonical Gs coupling to the prototypical Gi-coupled melatonin MT1 receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.08.687348v1?rss=1</link>
<description><![CDATA[
G protein-coupled receptors (GPCRs) transduce extracellular stimuli into intracellular signals by coupling to various heterotrimeric G proteins. However, the rules governing G protein preference remain largely elusive. MT1 and MT2 are prototypical Gi/o-coupled GPCRs responding to melatonin, a hormone secreted in a circadian manner. We show here that MT1, but not MT2, couples also to Gs proteins in vitro and activates the Gs/cAMP pathway upon long-term melatonin exposure in vivo, mimicking physiological dawn conditions. We solved the cryo-electron microscopy structure of the melatonin-MT1-Gs complex at 3.0[A] resolution, which revealed a strikingly distinct binding mode compared to the MT1-Gi complex. The third intracellular loop of MT1 emerges as a key stabilizer for Gs coupling, a feature previously unrecognized. This is the first solved receptor-Gs complex of a primary Gi-coupled GPCRs, providing new structural and functional insights into G protein selectivity and circadian switch of G protein coupling.
]]></description>
<dc:creator>Okamoto, H.</dc:creator>
<dc:creator>Oishi, A.</dc:creator>
<dc:creator>Ikegami, K.</dc:creator>
<dc:creator>McHugh, R.</dc:creator>
<dc:creator>Masri, B.</dc:creator>
<dc:creator>Kusakizako, T.</dc:creator>
<dc:creator>Kobayashi, K.</dc:creator>
<dc:creator>Karamitri, A.</dc:creator>
<dc:creator>Cecon, E.</dc:creator>
<dc:creator>Dam, J.</dc:creator>
<dc:creator>Nagase, M.</dc:creator>
<dc:creator>Tikhonova, I.</dc:creator>
<dc:creator>Nureki, O.</dc:creator>
<dc:creator>Jockers, R.</dc:creator>
<dc:date>2025-11-08</dc:date>
<dc:identifier>doi:10.1101/2025.11.08.687348</dc:identifier>
<dc:title><![CDATA[Structural basis and physiological significance of non-canonical Gs coupling to the prototypical Gi-coupled melatonin MT1 receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.07.687000v1?rss=1">
<title>
<![CDATA[
TriLeukeVax: A CD80/IL-15/IL-15Rα Expressing Autologous AML Cell Vaccine Elicits Robust Anti-Leukemic Cytolytic Activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.07.687000v1?rss=1</link>
<description><![CDATA[
Acute myeloid leukemia (AML) is the most common acute leukemia in adults and is associated with poor outcomes due to frequent relapse after remission induction. While hematopoietic stem cell transplantation (HSCT) can improve survival, many individuals, especially older patients, are ineligible. Prior immunotherapies have not reliably induced effective anti-leukemic immunity and have been associated with severe and unpredictable toxicities. Thus, there is a need for safe and effective therapies that reduce relapse and increase overall survival (OS). We have developed a universally applicable, patient-specific, lentivirally engineered autologous AML cell vaccine, TriLeukeVax (TLV), designed to stimulate leukemia-specific cytolytic immune responses in AML patients in remission. To generate TLV, AML cells are engineered to express the highly synergistic combination of the co-stimulatory protein CD80 and the IL-15/IL-15-receptor alpha (IL-15R) heterodimer. Prior proof-of-concept (POC) studies demonstrated eradication of disease in >80% of leukemic mice with serial administration of TLV. In the current studies, TLV was generated from 59/60 cryopreserved, diagnostic bone marrow-derived patient AML samples. Ex vivo priming of post-remission patient T-cells by ex vivo co-culture with autologous TLV stimulated robust proliferative and cytotoxic responses. In secondary co-cultures, T-cells previously primed by initial co-culture with TLV, showed greater clonal expansion and leukemia-specific cytolytic activity towards de novo autologous AML blasts than did control, unprimed T-cells. The enhanced anti-leukemic activity of TLV-primed T-cells against de novo AML confirms the potential for vaccine administration to effectively target minimal residual disease (MRD) persisting after chemotherapy and reduce relapse.

Key PointsO_LITriLeukeVax induces proliferation, activation, and effective anti-leukemic cytolytic responses in remission T-cells.
C_LIO_LIPrimed T-cells show polyclonal expansion and transcription profiles associated with proliferation, memory, and cytotoxicity.
C_LI



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=177 SRC="FIGDIR/small/687000v2_ufig1.gif" ALT="Figure UFIG">
View larger version (38K):
org.highwire.dtl.DTLVardef@5c1827org.highwire.dtl.DTLVardef@35b2e2org.highwire.dtl.DTLVardef@1732914org.highwire.dtl.DTLVardef@18df655_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOVisual Abstract.C_FLOATNO TriLeukeVax (TLV) workflow and mechanism of action.

Diagnostic AML patient bone marrow aspirates are collected, purified in Ficoll and cryopreserved. Samples are thawed and lentivirally transduced to produce TLV. Vaccination of AML patients in remission with TLV will stimulate the activation and expansion of leukemia-specific T-cells, effector memory cells, and NK cells by combining the co-stimulatory effects of CD80 with immune stimulation by the IL-15/IL-15R heterodimer expressed by the transduced AML cells, thereby targeting MRD and potentially increasing relapse-free survival in AML patients.

C_FIG
]]></description>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Wijayaratna, U. N.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Fung, J. P.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Farzaneh, F.</dc:creator>
<dc:creator>Kohn, D. B.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Gaensler, K. M. L.</dc:creator>
<dc:date>2025-11-09</dc:date>
<dc:identifier>doi:10.1101/2025.11.07.687000</dc:identifier>
<dc:title><![CDATA[TriLeukeVax: A CD80/IL-15/IL-15Rα Expressing Autologous AML Cell Vaccine Elicits Robust Anti-Leukemic Cytolytic Activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.08.687375v1?rss=1">
<title>
<![CDATA[
Parvalbumin interneurons gate and shape striatal sequences 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.08.687375v1?rss=1</link>
<description><![CDATA[
Loss of parvalbumin-expressing striatal interneurons (PV+) is associated with impulsive and uncontrolled behaviors. The underlying disruption in striatal information processing is unclear. We compared spiking of five identified neuron types in sensorimotor striatum, as unrestrained rats waited for a cue then performed brief, well-practiced actions. During waiting PV+ selectively increased firing, and suppressing PV+ at this time increased premature movements. This indicates a role in action restraint. However, suppressing PV+ after the cue had the opposite effect: slowed initiation and execution of actions. We found that each action was accompanied by a distinct, rapid striatal firing sequence, including both direct and indirect pathway spiny projection neurons (SPNs) and overlaid by sequential PV+ firing. Nearby pairs of PV+ and SPNs showed millisecond-level synchrony, and PV+ firing inhibited SPNs [~]2ms later. PV+ interneurons thus provide both broad restraint and precise sculpting of striatal output to achieve fluid, appropriately timed behavior.
]]></description>
<dc:creator>Duhne, M.</dc:creator>
<dc:creator>Gonzalez Montalvo, I.</dc:creator>
<dc:creator>Zheng, C. S.</dc:creator>
<dc:creator>Pelattini, L.</dc:creator>
<dc:creator>Berke, J. D.</dc:creator>
<dc:date>2025-11-09</dc:date>
<dc:identifier>doi:10.1101/2025.11.08.687375</dc:identifier>
<dc:title><![CDATA[Parvalbumin interneurons gate and shape striatal sequences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.07.687321v1?rss=1">
<title>
<![CDATA[
Dissecting mammalian cortical circuit development at single-cell resolution using inducible barcoded rabies virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.07.687321v1?rss=1</link>
<description><![CDATA[
Highly organized circuits of connected neurons enable diverse brain functions. Improper development of these circuits is associated with neurodevelopmental disorders, and understanding how circuits are formed is crucial for unraveling the mechanisms of these diseases. We currently have an incomplete picture of how specific brain circuits develop and how they are affected in disease, because we lack methods to study them at scale and with single-cell resolution. Monosynaptic rabies tracing is the gold standard method to study circuit architecture. However, it suffers from cellular toxicity, low throughput, lack of control over the timing of labeling, and the inability to access the molecular profiles of individual neurons. To address these issues, we developed an inducible barcoded rabies virus (ibRV) to enable temporal-controlled labeling of synaptic circuits followed by high-throughput single-cell genomics readout. ibRV allows for dissecting neuronal circuit changes over time at single-cell and spatial resolution. We applied ibRV to study the development of specific mouse cortical circuits during late prenatal and postnatal life using single-cell genomics and unbiased spatial transcriptomics as readouts. We characterized and quantified developmental connectivity patterns and molecular cascades that underlie their formation. Additionally, we constructed functional in silico circuit models that enable interrogation of circuit function and dysfunction at specific developmental stages. Our study provides novel tools for circuit analysis and can provide new insights into the mechanisms of mammalian brain development.
]]></description>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>D'Arcy, B.</dc:creator>
<dc:creator>He, L.</dc:creator>
<dc:creator>Arroyo, D.</dc:creator>
<dc:creator>Deasy, S.</dc:creator>
<dc:creator>Matthews, E.</dc:creator>
<dc:creator>Yan, Z.</dc:creator>
<dc:creator>Rastogi, N.</dc:creator>
<dc:creator>Mancia Leon, W.</dc:creator>
<dc:creator>Kaur, K. S.</dc:creator>
<dc:creator>Southwell, D. G.</dc:creator>
<dc:creator>Huang, Z. J.</dc:creator>
<dc:creator>Silver, D.</dc:creator>
<dc:creator>Velmeshev, D.</dc:creator>
<dc:date>2025-11-10</dc:date>
<dc:identifier>doi:10.1101/2025.11.07.687321</dc:identifier>
<dc:title><![CDATA[Dissecting mammalian cortical circuit development at single-cell resolution using inducible barcoded rabies virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.10.687296v1?rss=1">
<title>
<![CDATA[
A conserved sRNA regulates mucin adhesion and gut colonization across the Enterococcaceae 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.10.687296v1?rss=1</link>
<description><![CDATA[
Enterococci, particularly E. faecalis, can survive in diverse settings within and outside human hosts. The capacity of E. faecalis to colonize these locations relies on its ability to adapt by altering gene expression in response to environmental exposures. One mechanism for quickly altering gene expression is through regulation by small noncoding RNAs (sRNAs); sRNAs can regulate one or many target genes and either up- or down-regulate transcript stability and protein expression. While many sRNAs have been predicted in E. faecalis, few have experimentally established target mRNAs or physiological functions. Here, we investigate the targets, function, and mechanism of Enterococcus sRNA 84. We found that sRNA 84 is conserved within the family Enterococcaceae, suggesting that it plays a role in the regulation of core genes and functions. RNA sequencing and proteomic analysis revealed that the absence of sRNA 84 led to downregulation of many cell surface proteins, including mucin-binding proteins. Consistent with these findings, an sRNA 84 knockout strain had reduced binding to mucin in vitro and impaired intestinal colonization of specific-pathogen-free mice. Taken together, these data support a model whereby sRNA 84 upregulates cell surface adhesins, which subsequently facilitate host colonization through binding to mucin. sRNA 84 is one of the first sRNAs in enterococci with demonstrated targets and function. This finding establishes the conserved sRNA 84 as a potential key regulator of enterococcal host adaptation, providing insight into how these organisms adapt their gene expression to survive both within and outside animal hosts.
]]></description>
<dc:creator>Bowden, S.</dc:creator>
<dc:creator>Jennings, K. C.</dc:creator>
<dc:creator>Zlitni, S.</dc:creator>
<dc:creator>Brooks, E. F.</dc:creator>
<dc:creator>Davin, M. E.</dc:creator>
<dc:creator>Natarajan, A.</dc:creator>
<dc:creator>Asare, L.</dc:creator>
<dc:creator>Fremin, B. J.</dc:creator>
<dc:creator>Wilmes, P.</dc:creator>
<dc:creator>Hettich, R. L.</dc:creator>
<dc:creator>Salzman, N. H.</dc:creator>
<dc:creator>Bhatt, A. S.</dc:creator>
<dc:date>2025-11-11</dc:date>
<dc:identifier>doi:10.1101/2025.11.10.687296</dc:identifier>
<dc:title><![CDATA[A conserved sRNA regulates mucin adhesion and gut colonization across the Enterococcaceae]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.11.687767v1?rss=1">
<title>
<![CDATA[
Scalable imaging-based profiling of CRISPR perturbations with protein barcodes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.11.687767v1?rss=1</link>
<description><![CDATA[
Imaging-based CRISPR screens enable high-content functional genomics by capturing phenotypic changes in cells after genetic perturbation. Protein barcodes provide cost-effective, easy-to-implement, and imaging-compatible barcoding for pooled perturbations, yet their scalability has been constrained by the need for arrayed cloning, lentiviral recombination between barcodes and guides, and difficulties in decoding barcodes with high confidence. Here, we introduce poolVis and cellPool, an integrated experimental and computational platform designed to address these limitations. poolVis uses Cre-lox-mediated reconfiguration to position barcode-sgRNA pairs in proximity during viral integration, which greatly reduces barcode shuffling during pooled cloning and delivery. cellPool leverages a scalable computational workflow and the unique aspects of protein barcodes to produce unpooled image galleries from multi-terabyte scale datasets. Applying this platform to single- and double-CRISPRi profiling of cell-cycle genes and chromokinesins in the MCF10A cells uncovered established and previously unrecognized phenotypes, including nuclear morphology changes and reciprocal sign epistasis in DNA damage.
]]></description>
<dc:creator>Choudhary, K.</dc:creator>
<dc:creator>McManus, M.</dc:creator>
<dc:date>2025-11-11</dc:date>
<dc:identifier>doi:10.1101/2025.11.11.687767</dc:identifier>
<dc:title><![CDATA[Scalable imaging-based profiling of CRISPR perturbations with protein barcodes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.10.687749v1?rss=1">
<title>
<![CDATA[
Lactate dehydrogenase-induced DNA Topoisomerase 1 is a novel regulator of smooth muscle cell proliferation and remodeling in pulmonary arterial hypertension 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.10.687749v1?rss=1</link>
<description><![CDATA[
Pulmonary arterial hypertension (PAH) manifests by increased proliferation and survival of pulmonary vascular cells in small pulmonary arteries (PAs), PA remodeling and unresolved increase of PA pressure. PA smooth muscle cells (PASMCs) in PAH undergo metabolic shift to glycolysis resulting in over-production of lactate, hyper-proliferation, and apoptosis resistance, but the mechanisms are not completely understood. By using lung tissues and pulmonary vascular cells from PAH and non-diseased human lungs, unbiased proteomics, network analysis, and gain-and-loss of function approaches, we here report that up-regulation of lactate dehydrogenase A (LDHA)-lactate axis promotes PASMC-specific over-lactylation and consequent over-accumulation of DNA topoisomerase 1 (TOP1) in small remodeled PAs from PAH lungs, leading to the up-regulation of Akt-mechanistic target of rapamycin 1 (mTORC1) signaling, hyper-proliferation, and reduced apoptosis. Smooth muscle-specific LDHA knockdown prevented, and Ldha inhibitor oxamate reversed SU5416/hypoxia-induced TOP1 accumulation, pulmonary vascular remodeling, and pulmonary hypertension (PH) in mice. Pharmacological inhibition of TOP1 with indotecan suppressed Akt-mTORC1, decreased proliferation, induced apoptosis in human PAH, but not control PASMCs, and reversed PA remodeling, PH, and RV dysfunction in rats. Collectively, these data provide a novel mechanistic link from LDHA-driven lactate over-production through lactylation and overaccumulation of TOP1, to the up-regulation of Akt-mTORC1, hyper-proliferation and apoptosis resistance of PASMCs, pulmonary vascular remodeling, and PH, and identify TOP1 as a new potentially attractive molecular target for the remodeling-focused therapeutic intervention.

Take-home messageLDHA-lactate-induced over-lactylation and overaccumulation of Topoisomerase 1 (TOP1) promotes pulmonary artery smooth muscle cell hyper-proliferation, remodeling, and pulmonary arterial hypertension, which are reversed by TOP1 inhibitor indotecan.
]]></description>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>Zhyvylo, I.</dc:creator>
<dc:creator>Goncharov, D.</dc:creator>
<dc:creator>Dey, T.</dc:creator>
<dc:creator>Kelly, N.</dc:creator>
<dc:creator>Teos, L.</dc:creator>
<dc:creator>Franzi, L.</dc:creator>
<dc:creator>Saiyed, A.</dc:creator>
<dc:creator>Kenyon, N. J.</dc:creator>
<dc:creator>Greenland, J. R.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:creator>Chen, S. Y.</dc:creator>
<dc:creator>Delisser, H.</dc:creator>
<dc:creator>Kudryashova, T. V.</dc:creator>
<dc:creator>Goncharova, E. A.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.10.687749</dc:identifier>
<dc:title><![CDATA[Lactate dehydrogenase-induced DNA Topoisomerase 1 is a novel regulator of smooth muscle cell proliferation and remodeling in pulmonary arterial hypertension]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.10.687758v1?rss=1">
<title>
<![CDATA[
Putative azithromycin-resistance mutations in Chlamydia trachomatis are globally distributed but arose before azithromycin was discovered 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.10.687758v1?rss=1</link>
<description><![CDATA[
Azithromycin is widely used to treat Chlamydia trachomatis infections, yet the extent of resistance to the drug across the species has not been addressed. We surveyed mutations and substitutions linked to putative azithromycin resistance across 1,354 high-quality C. trachomatis genomes. Mutations in the rplV gene encoding three non-synonymous substitutions, compared to the canonical C. trachomatis reference strain D//TW-3/Cx sequence, were found to be common but largely conserved within lineages causing prevalent urogenital and anorectal infections and lymphogranuloma venereum (LGV). Time-scaled phylogenetic analysis suggested that these mutations predate the clinical introduction of azithromycin. In contrast, no consistent resistance-associated patterns were observed in 23S rRNA or rplD genes. This large-scale genomic surveillance provides critical insights into the evolutionary trends of putative azithromycin resistance in C. trachomatis and underscores the importance of integrating genomic monitoring with phenotypic susceptibility testing to accurately assess and manage antimicrobial resistance.
]]></description>
<dc:creator>Sharma, P.</dc:creator>
<dc:creator>Dean, D.</dc:creator>
<dc:creator>Read, T. D.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.10.687758</dc:identifier>
<dc:title><![CDATA[Putative azithromycin-resistance mutations in Chlamydia trachomatis are globally distributed but arose before azithromycin was discovered]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.11.687663v1?rss=1">
<title>
<![CDATA[
Fast-conducting mechanonociceptors uniquely engage reflexive and affective pain circuitry to drive protective responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.11.687663v1?rss=1</link>
<description><![CDATA[
Nociceptors detect damaging stimuli and evoke pain in healthy animals. We conducted an optogenetic activation screen to identify genetically defined nociceptor populations that elicit place aversion and nocifensive behaviors in response to stimulation. Smr2Cre- and Bmpr1bCre-labeled A{delta} high-threshold mechanoreceptors (HTMRs) emerged as two of the few nociceptor populations, and we focused on investigating their physiological, morphological, functional, and synaptic properties. These neurons densely innervate skin and other organs, are activated only by intense, potentially damaging stimuli, and are necessary for protective responses to sharp mechanical stimuli. Centrally, A{delta}-HTMRs projections span multiple spinal segments and terminate across spinal cord laminae, forming strong, monosynaptic connections onto anterolateral tract projection neurons, including antenna cells of the deep dorsal horn. A{delta}-HTMRs also engage a local spinal reflex circuit enabling a remarkably rapid limb withdrawal. Thus, A{delta}-HTMRs are myelinated nociceptors with unique properties that can be exploited for development of new analgesics.
]]></description>
<dc:creator>Lezgiyeva, K.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Nguyen, K.</dc:creator>
<dc:creator>DeLisle, M. M.</dc:creator>
<dc:creator>Ko, F. C.</dc:creator>
<dc:creator>Fullam, S.</dc:creator>
<dc:creator>Obeidat, A. M.</dc:creator>
<dc:creator>Turecek, J.</dc:creator>
<dc:creator>Alkislar, I.</dc:creator>
<dc:creator>Lehnert, B. P.</dc:creator>
<dc:creator>Martinez-Garcia, R. I.</dc:creator>
<dc:creator>Sivakumar, R.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Mazor, O.</dc:creator>
<dc:creator>Garibyan, L.</dc:creator>
<dc:creator>Sharma, N.</dc:creator>
<dc:creator>Emanuel, A. J.</dc:creator>
<dc:creator>Malfait, A.-M.</dc:creator>
<dc:creator>Miller, R. E.</dc:creator>
<dc:creator>Ginty, D. D.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.11.687663</dc:identifier>
<dc:title><![CDATA[Fast-conducting mechanonociceptors uniquely engage reflexive and affective pain circuitry to drive protective responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.11.687887v1?rss=1">
<title>
<![CDATA[
APOE4-Aβ synergy drives brain network dysfunction and neuronal lysosomal-ER proteostasis dysregulation in preclinical Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.11.687887v1?rss=1</link>
<description><![CDATA[
Amyloid-{beta} (A{beta}) and APOE4 represent two of the strongest pathological and genetic risk factors for Alzheimers disease (AD), but how these co-pathogens interact during preclinical stages remains undefined. We addressed this question by developing a humanized knock-in model expressing physiological, endogenously regulated human A{beta} and APOE4. Aged AppNLF:APOE4 mice displayed incipient amyloidosis with subtle memory-related changes, consistent with preclinical AD. We found largely distinct, non-overlapping APOE4- and A{beta}-driven functional synaptic, sleep, and behavioral alterations. However, at the transcriptomic level, APOE4xA{beta} had a pronounced detrimental interaction in neuronal populations, whereas glial populations were primarily affected by either genotype. We found APOE4xA{beta} molecular interactions in neuronal populations, including excitatory and inhibitory cells, converged on a core lysosomal-ER proteostasis axis. We propose that APOE4xA{beta} interaction produces an early neuronal pathogenic signature, involving the lysosomal-ER proteostasis axis, preceding functional decline and driving disease progression. APOE4xA{beta}-KI models provide a physiologically relevant platform to study early pathogenesis.

HighlightsO_LIEarly synergistic APOE4xA{beta} interaction emerges predominantly at the transcriptomic level in neurons, but not in glial cells.
C_LIO_LIAPOE4 and A{beta} drive largely non-overlapping physiological changes in preclinical stages of disease, but converge at the level of network hyperexcitability.
C_LIO_LIAPOE4xA{beta} neuronal synergy converges on a conserved lysosomal-ER proteostasis axis.
C_LIO_LIHumanized APOE4xA{beta} KI mice provide a physiologically relevant model to dissect early AD pathogenesis in preclinical stages
C_LI
]]></description>
<dc:creator>Shin, J.</dc:creator>
<dc:creator>Brady, E.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Lauderdale, K.</dc:creator>
<dc:creator>Agrawal, A.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Jiang, X.</dc:creator>
<dc:creator>Herbert, J.</dc:creator>
<dc:creator>Mallen, D. D.</dc:creator>
<dc:creator>Ly, K. K.</dc:creator>
<dc:creator>Honma, P. S.</dc:creator>
<dc:creator>Guo, Z.</dc:creator>
<dc:creator>Sant, C.</dc:creator>
<dc:creator>Thomas, R.</dc:creator>
<dc:creator>Miller, S. R.</dc:creator>
<dc:creator>Cobos, I.</dc:creator>
<dc:creator>Palop, J. J.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.11.687887</dc:identifier>
<dc:title><![CDATA[APOE4-Aβ synergy drives brain network dysfunction and neuronal lysosomal-ER proteostasis dysregulation in preclinical Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.11.687898v1?rss=1">
<title>
<![CDATA[
Co-Hyperpolarized Dehydroascorbate and Pyruvate MRI Predicts Radiation Response in Glioblastoma Models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.11.687898v1?rss=1</link>
<description><![CDATA[
PurposeEarly noninvasive assessments of treatment response are desperately needed to improve outcomes in glioblastoma (GBM). Molecular imaging techniques that measure glycolytic metabolism are being increasingly studied, but limitations such as variable substrate delivery present significant barriers to clinical interpretation. To develop more robust translational imaging biomarkers, we propose utilizing the interrogation of oxidative stress, a critical component of tumor metabolism for which no method of clinical measurement currently exists. This study investigates the simultaneous measure of oxidative stress and glycolytic flux using co-hyperpolarized [1-13C] dehydroascorbate and [1-13C] pyruvate (HP DHA/PA) as a predictor of treatment response in GBM.

Experimental DesignTo establish a model that exhibits known metabolic responses to oxidative stress, we characterize radiation induced metabolic reprogramming in four human GBM lines (U87, U251, A172, T98) in vitro. We extend this in vivo and establish radiosensitive and radioresistant orthotopic xenograft models to investigate HP DHA/PA magnetic resonance imaging as a predictor of treatment response.

ResultsIn vitro analyses revealed that radiation upregulates the pentose phosphate pathway and response is augmented by glutathione depletion. In vivo metabolomic profiling identified preferential nucleotide metabolism pathways in each tumor type. HP DHA/PA imaging revealed that DHA perfusion was not impacted by blood-brain-barrier integrity and detected reductions in DHA-to-vitamin C and pyruvate-to-lactate conversion in treatment-sensitive tumors, reflecting diminished reductive capacity following radiation.

ConclusionsThese findings demonstrate successful prediction of radiosensitivity in GBM utilizing measurement of oxidative stress and establish HP DHA/PA imaging as an innovative method to address existing clinical limitations in treatment response assessment.
]]></description>
<dc:creator>Coffee, E.</dc:creator>
<dc:creator>Porcari, P.</dc:creator>
<dc:creator>Patel, S.</dc:creator>
<dc:creator>Figlioli, G.</dc:creator>
<dc:creator>Berishaj, M.</dc:creator>
<dc:creator>Rahimi-Keshari, K.</dc:creator>
<dc:date>2025-11-13</dc:date>
<dc:identifier>doi:10.1101/2025.11.11.687898</dc:identifier>
<dc:title><![CDATA[Co-Hyperpolarized Dehydroascorbate and Pyruvate MRI Predicts Radiation Response in Glioblastoma Models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.12.688091v1?rss=1">
<title>
<![CDATA[
Class-I myosin responds to change in membrane tension during clathrin-mediated endocytosis in human induced pluripotent stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.12.688091v1?rss=1</link>
<description><![CDATA[
Clathrin-mediated endocytosis (CME) is an essential cellular process that needs to operate efficiently across a wide range of conditions. Internalization of the endocytic site involves forces generated by membrane-bound proteins and Arp2/3-mediated branched actin filament assembly to bend the plasma membrane (PM) from flat to omega-shaped. In mammalian CME, the requirement for a branched actin filament network varies depending on cell type and differences in membrane tension. However, how the actin network adapts to changes in load in order to ensure robustness of this process over a range of membrane tensions is not understood. Here, we combine live-cell imaging and super-resolution microscopy of genome-edited human induced pluripotent stem cells (hiPSCs) to investigate the role of the mammalian class-I myosin, Myosin1E (Myo1E), in load adaptation. Under normal conditions, sites that recruit Myo1E are rare and exhibit slow CME dynamics. However, as membrane tension increases and CME dynamics are slowed globally, Myo1E is recruited to more sites, likely to assemble more branched actin, resulting in increased force generation to rescue stalled sites and promote internalization. Loss of Myo1E results in increased Arp2/3 complex lifetime at CME sites under normal conditions, and at high membrane tension, these sites fail to recruit as many Arp2/3 molecules. We propose that Myo1E is recruited to CME sites that have stalled due to increased membrane tension, where it helps build a more effective branched actin network by generating force through motor activity and recruiting additional Arp2/3 complexes to rescue stalled sites.

SignificanceFor mammalian cells to internalize extracellular cargo via clathrin-mediated endocytosis (CME), specific regions of the plasma membrane (PM) must bend from flat to inwardly curved, a process that requires force-generating proteins. One key component in generating this force during CME is the branched actin network, in which actin filaments polymerize against the plasma membrane. When PM tension increases, more force is required to generate curvature, prompting the assembly of actin and actin associated proteins to aid the process. We demonstrate that the class-I myosin motor, Myosin1E (Myo1E), becomes increasingly crucial as membrane tension rises, presumably to build a more effective branched actin network to facilitate internalization of slowed sites.
]]></description>
<dc:creator>Smith, S. L.</dc:creator>
<dc:creator>Zhan, T.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>De Belly, H.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Xu, K.</dc:creator>
<dc:creator>Weiner, O.</dc:creator>
<dc:creator>Drubin, D. G.</dc:creator>
<dc:date>2025-11-13</dc:date>
<dc:identifier>doi:10.1101/2025.11.12.688091</dc:identifier>
<dc:title><![CDATA[Class-I myosin responds to change in membrane tension during clathrin-mediated endocytosis in human induced pluripotent stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.12.688154v1?rss=1">
<title>
<![CDATA[
Pre-existing and emergent cortical neuronal assembly sequences during learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.12.688154v1?rss=1</link>
<description><![CDATA[
Neuronal assemblies -- groups of co-active neurons -- support memory consolidation and retrieval. In the hippocampus, assemblies can pre-exist learning and contribute to memory through sequential activation. Whether similar principles apply to higher cortical areas for flexible memory storage like the orbitofrontal cortex (OFC) remains unclear. Using a novel ground truth-validated clustering approach, we investigated the activity of longitudinally tracked mouse OFC neurons during cue-reward memory acquisition and maintenance. Assemblies active after learning pre-existed the learning and exhibited two distinct sequential dynamics consistent with memory consolidation or retrieval. Consolidation sequences emerged during learning, while retrieval sequences partly recruited pre-existing reward sequences. These findings demonstrate that OFC learning recruits pre-existing networks flexibly repurposed for new associations, revealing circuit motifs that may enable cortical memory storage.
]]></description>
<dc:creator>Pemberton, L.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>K Namboodiri, V. M.</dc:creator>
<dc:date>2025-11-13</dc:date>
<dc:identifier>doi:10.1101/2025.11.12.688154</dc:identifier>
<dc:title><![CDATA[Pre-existing and emergent cortical neuronal assembly sequences during learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.13.688312v1?rss=1">
<title>
<![CDATA[
Acute Myeloid Leukemia Relapse after Bromodomain Inhibitor Treatment or Chemotherapy is Characterized by Myc-Ras Transcriptional Remodeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.13.688312v1?rss=1</link>
<description><![CDATA[
Adult and pediatric acute myeloid leukemias (AMLs) harbor distinct mutational profiles, including a higher incidence of RAS and other signaling mutations in young patients. Here we show that the BET inhibitor PLX51107 potently suppresses the growth of NRAS-mutant AML cell lines, and that these activities are enhanced by co-treatment with the MEK inhibitor PD0325901. Controlled preclinical trials in primary mouse Nras-mutant AMLs revealed single agent efficacy of PLX51107 that was enhanced by PD0325901. Leukemias that relapsed during treatment developed intrinsic drug resistance characterized by transition to a more primitive state, up-regulation of Myc target genes, and down-regulation of Ras-associated transcriptional programs. AMLs that relapsed after frontline chemotherapy showed similar transcriptional remodeling. These studies demonstrate transcriptional plasticity in primary AMLs that relapse following in vivo treatment with either targeted agents or chemotherapy, and support evaluating BET inhibition in leukemias with monocytic differentiation and RAS mutations.
]]></description>
<dc:creator>Huang, B. J.</dc:creator>
<dc:creator>D'Souza, J.</dc:creator>
<dc:creator>Batingana, A. R.</dc:creator>
<dc:creator>Harris, M. D.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Hwang, E.</dc:creator>
<dc:creator>Wandler, A. M.</dc:creator>
<dc:creator>Burgess, M. R.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Bollag, G.</dc:creator>
<dc:creator>Shannon, K.</dc:creator>
<dc:date>2025-11-14</dc:date>
<dc:identifier>doi:10.1101/2025.11.13.688312</dc:identifier>
<dc:title><![CDATA[Acute Myeloid Leukemia Relapse after Bromodomain Inhibitor Treatment or Chemotherapy is Characterized by Myc-Ras Transcriptional Remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.13.688348v1?rss=1">
<title>
<![CDATA[
Decoding hidden goal-directed navigational states and their neuronal representations using a novel labyrinth paradigm and probabilistic modeling framework 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.13.688348v1?rss=1</link>
<description><![CDATA[
Goal-directed navigation involves a sequence of planned actions aimed at achieving long-term goals through reinforcement, but detecting hidden states that support this process and their neuronal substrates remains a fundamental challenge. To address this, we developed a complex labyrinth test that mimics naturalistic foraging and implemented a novel hierarchical probabilistic modeling framework, Cognitive Mapping of Planned Actions with State Spaces (CoMPASS). This framework infers a nested state structure, comprising short-term surveillance-ambulation states (Level 1) and long-term goal-oriented navigational states (Level 2). Using CoMPASS, we show that successful navigation in wild-type mice is marked by increased recruitment of both surveillance and goal-oriented states specifically at decision nodes, revealing how sequential behavioral decisions culminate in long-term goals. In contrast, the humanized AppSAA mouse model of Alzheimers disease (AD) exhibited navigational impairments marked by diminished surveillance during decisions, reduced goal-directed states, and increased navigation stochasticity. Importantly, we show that gamma oscillations in the posterior parietal cortex (PPC), a region involved in spatial navigation planning, encode these CoMPASS behavioral states and their dynamic operating modes linking spatial locations to long-term goals. Our findings provide a novel paradigm for assessing hidden goal-directed navigational states and identify gamma oscillations in the PPC as their neural substrates.
]]></description>
<dc:creator>Honma, P. S.</dc:creator>
<dc:creator>Bangera, S. C.</dc:creator>
<dc:creator>Thomas, R. R.</dc:creator>
<dc:creator>Kaliss, N.</dc:creator>
<dc:creator>Xia, D.</dc:creator>
<dc:creator>Palop, J. J.</dc:creator>
<dc:date>2025-11-14</dc:date>
<dc:identifier>doi:10.1101/2025.11.13.688348</dc:identifier>
<dc:title><![CDATA[Decoding hidden goal-directed navigational states and their neuronal representations using a novel labyrinth paradigm and probabilistic modeling framework]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.14.688347v1?rss=1">
<title>
<![CDATA[
The phenotypic landscape of the mycobacterial cell 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.14.688347v1?rss=1</link>
<description><![CDATA[
The Mycobacteriales are an order of diverse bacteria that thrive in many environmental and host-associated niches. Because the most notorious member of this clade, Mycobacterium tuberculosis, is a major human pathogen, research on Mycobacteriales has focused on pathogenesis, and, as a consequence, many fundamental aspects of Mycobacterial biology remain understudied. Here, we address this gap by performing a genome-wide CRISPRi chemical genomics screen using a diverse set of >35 antibiotics, detergents, and other anti-microbials predominantly targeting the cell envelope of Mycobacterium smegmatis, a saprophytic model Mycobacterium. We highlight new information derived from this screen, including the identification of novel functions for previously uncharacterized conserved and essential genes (in mycolic acid and arabinogalactan synthesis), the discovery of a new drug scaffold/protein target pair, and insights into the mechanism of action of two commonly used antibiotics. These data are also a valuable resource for the mycobacterial research community, as they provide thousands of novel phenotypes for uncharacterized genes and meaningful phenotypic correlations between annotated and uncharacterized genes.
]]></description>
<dc:creator>Herrera, N.</dc:creator>
<dc:creator>Todor, H.</dc:creator>
<dc:creator>Kim, L. M.</dc:creator>
<dc:creator>Burkhart, H. N.</dc:creator>
<dc:creator>Billings, E.</dc:creator>
<dc:creator>Warner, T. C.</dc:creator>
<dc:creator>Lee, S. Y.</dc:creator>
<dc:creator>Sayegh, N. Y.</dc:creator>
<dc:creator>Bosch, B.</dc:creator>
<dc:creator>Kiessling, L. L.</dc:creator>
<dc:creator>Mancia, F.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:creator>Gross, C. A.</dc:creator>
<dc:date>2025-11-15</dc:date>
<dc:identifier>doi:10.1101/2025.11.14.688347</dc:identifier>
<dc:title><![CDATA[The phenotypic landscape of the mycobacterial cell]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.17.688783v1?rss=1">
<title>
<![CDATA[
Model of ocular surface ion and water transport predicts efficacy of dry eye therapeutics targeting epithelial transport and tear fluid dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.17.688783v1?rss=1</link>
<description><![CDATA[
The composition and volume of tear fluid lining the ocular surface are governed by the rates of lacrimal gland secretion, tear film evaporation, nasolacrimal drainage, and epithelial ion and water transport. Tear fluid hyperosmolality and reduced volume are key drivers of dry eye disease (DED) pathogenesis. We constructed a mathematical model to compute the composition and volume of tear fluid and epithelial cell compartments, with transport parameters specified for the mouse eye from published data and in vivo measurements of ocular surface potential differences. The model accounted for transcellular and paracellular transport across the epithelia under open-circuit conditions utilizing flux equations for individual transport mechanisms, with mass balance constraints on solute and water content in cytoplasm and tear fluid. Under DED conditions established by reduced lacrimal secretion and increased evaporation, the model predicted the efficacy of currently available DED therapies including punctal plugs, humidification goggles, lacrimal gland stimulation, and artificial tears eye drops. The model also predicted the limited efficacy of anti-absorptive and pro-secretory drugs targeting epithelial ion transporters, and the high efficacy of targeting epithelial water permeability or paracellular ion permeability. The modeling herein provided quantitative predictions to prioritize novel targets for DED and drive the development of new therapies.

Author SummaryDry eye disease (DED) affects billions of adults worldwide, but a full picture is lacking of how the tear film becomes abnormally thin and hyperosmolar. The computer model built here links four processes - tear production by the lacrimal gland, tear fluid evaporation, tear drainage through tear ducts, and transport of solutes and water across eye surface epithelial cells - to predict the thickness and composition of the tear film in various conditions. Model parameters were selected using published data and electrical measurements of voltage changes across the ocular surface produced by ion transport. The model predicted that existing therapies, such as punctal plugs, moisture goggles, or stimulating tear production, can substantially increase tear thickness or lower saltiness. The model also predicted limited efficacy of drug therapies in current development that target ion transport, and identified epithelial cell water transport and paracellular ion permeability as novel targets for DED treatment. By making ocular surface transport mechanisms explicit and testable, our work offers a roadmap for development of new therapies that restore a healthy tear film, a major unmet medical need.
]]></description>
<dc:creator>Verma, V.</dc:creator>
<dc:creator>Lindgren, E. S.</dc:creator>
<dc:creator>Levin, M. H.</dc:creator>
<dc:creator>Cil, O.</dc:creator>
<dc:creator>Tradtrantip, L.</dc:creator>
<dc:creator>Yan, R.</dc:creator>
<dc:creator>Pasricha, N. D.</dc:creator>
<dc:creator>Verkman, A. S.</dc:creator>
<dc:date>2025-11-17</dc:date>
<dc:identifier>doi:10.1101/2025.11.17.688783</dc:identifier>
<dc:title><![CDATA[Model of ocular surface ion and water transport predicts efficacy of dry eye therapeutics targeting epithelial transport and tear fluid dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.17.688886v1?rss=1">
<title>
<![CDATA[
TAF15 amyloids propagate via defined motifs in a prion-like fashion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.17.688886v1?rss=1</link>
<description><![CDATA[
TATA-box binding protein-associated factor 15 (TAF15) is an RNA-binding member of the FET family recently identified as the primary fibrillar constituent in a subset of frontotemporal lobar degeneration (FTLD-FET) cases. Although TAF15 is also linked to amyotrophic lateral sclerosis (ALS), the molecular basis and propagation behavior of its aggregates remain unknown. In this work, we show that recombinant TAF15 forms amyloid fibrils under physiological conditions and developed a single-fluorophore TAF15 biosensor to quantitatively monitor their cellular propagation. Using this system, we demonstrate that both recombinant TAF15 fibrils and pathological aggregates extracted from atypical FTLD with ubiquitin inclusions (aFTLD-U) patient brains seed aggregation efficiently and transmit serially between cells, demonstrating hallmark features of prion-like propagation. Seeding was specific to TAF15 and absent for other amyloidogenic proteins, including the homologous protein fused-in-sarcoma (FUS), revealing an unexpected cross-seeding barrier. Occasional colocalization of FUS within TAF15 inclusions was observed upon transient co-expression, suggesting that FUS can be passively recruited rather than acting as an inducer of pathology in FTLD-FET brains. Computational and peptide-based experimental mapping identified multiple aggregation-prone regions within the TAF15 low-complexity domain that coincide with hotspots stabilizing the core of ex vivo TAF15 amyloid fibrils. These short motifs encode the propagation propensity of TAF15 aggregation in vitro and in cells. Together, these findings establish TAF15 as a bona fide amyloid-forming, prion-like protein and define the sequence grammar underlying its self-assembly, providing a mechanistic framework for its role in FTLD-FET and ALS and offering tractable molecular targets for therapeutic intervention.
]]></description>
<dc:creator>Konstantoulea, K.</dc:creator>
<dc:creator>Gadhe, L.</dc:creator>
<dc:creator>Goodavish, F.</dc:creator>
<dc:creator>GUPTA, A.</dc:creator>
<dc:creator>Tagad, H.</dc:creator>
<dc:creator>Vaquer-Alicea, J.</dc:creator>
<dc:creator>Ghayal, N. B.</dc:creator>
<dc:creator>Roemer, S. F.</dc:creator>
<dc:creator>DeTure, M. A.</dc:creator>
<dc:creator>Nana, A. L.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Dickson, D. W.</dc:creator>
<dc:creator>White, C. L.</dc:creator>
<dc:creator>Diamond, M. I.</dc:creator>
<dc:creator>Louros, N. N.</dc:creator>
<dc:date>2025-11-18</dc:date>
<dc:identifier>doi:10.1101/2025.11.17.688886</dc:identifier>
<dc:title><![CDATA[TAF15 amyloids propagate via defined motifs in a prion-like fashion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.18.689137v1?rss=1">
<title>
<![CDATA[
Scalable Human Cellular Models of Parkinson Disease Reveal A Druggable Link Between the Angiotensin Receptor 1 and Alpha-Synuclein Pathology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.18.689137v1?rss=1</link>
<description><![CDATA[
BackgroundParkinsons disease (PD) involves progressive loss of midbrain dopaminergic (mDA) neurons in the substantia nigra. No disease-modifying treatments exist, only symptomatic relief. Our lab reported an unbiased screen in larval zebrafish identifying renin-angiotensin-aldosterone system (RAAS) inhibitors, including clinically used AGTR1 inhibitors for hypertension, as potent neuroprotective agents. This study aims to investigate the effects of AGTR1 inhibition on human mDA neuron survival using inducible neurodegenerative 2D and 3D models for human mDA neuron degeneration.

MethodsWe report a scalable high-content platform, using CRISPR-engineered human induced pluripotent stem cell (hiPSC)-derived mDA neurons expressing a tyrosine hydroxylase (TH) fluorescent reporter, allowing to track mDA neuron survival live "in a dish". We developed chemically inducible neurodegenerative 2D and 3D models for human mDA neuron degeneration, allowing to recapitulate PD pathology in human cells in vitro.

ResultsOur model establishes scalable human cellular models of PD well-suited for therapeutic discovery. Using 2D and 3D mono and co-cultures, this study demonstrates that inhibition of AGTR1, via chemical or genetic means, protects against chemically induced mDA neuron degeneration. Transcriptomic analyses show AGTR1 inhibition lowers synuclein transcription, by reducing SNCA and SNCB gene expression. In 3D neuron-glia assembloids, AGTR1 inhibition protects against the accumulation of phosphorylated form of -synuclein (p129-Syn), key PD pathological marker.

ConclusionsWe highlight AGTR1 as a key regulator of -synuclein transcription and aggregation in human mDA neurons, and AGTR1 inhibition as pro-survival in human iPSC-derived models. These findings position inhibition of AGTR1 as a promising therapeutic strategy for PD neuroprotection.
]]></description>
<dc:creator>Daynac, M.</dc:creator>
<dc:creator>Mouilleau, V.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:date>2025-11-19</dc:date>
<dc:identifier>doi:10.1101/2025.11.18.689137</dc:identifier>
<dc:title><![CDATA[Scalable Human Cellular Models of Parkinson Disease Reveal A Druggable Link Between the Angiotensin Receptor 1 and Alpha-Synuclein Pathology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.20.689476v1?rss=1">
<title>
<![CDATA[
Harmonized Protocol for Segmentation of the Hippocampal Tail on High-Resolution in vivo MRI from the Hippocampal Subfields Group (HSG) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689476v1?rss=1</link>
<description><![CDATA[
The hippocampus is a heterogeneous structure with cytoarchitectonically distinct subfields that exhibit heterogeneous lifespan trajectories and are differentially susceptible to diseases. Advances in high-resolution imaging have accelerated research on these structures, yet variability in segmentation protocols limits cross-study comparability. The Hippocampal Subfields Group (HSG) is an international consortium addressing this challenge by developing a reliable, accessible, and freely available segmentation protocol for high-resolution T2-weighted 3 tesla MRI scans (http://www.hippocampalsubfields.com). Here, we present the harmonized protocol for the posterior portion of the hippocampus (the "tail"), complementing the previously established "body" protocol, and with an anterior "head" protocol under development. The tail protocol provides standardized definitions of the external boundaries for the posterior-most extent of the hippocampus, facilitating consistent segmentation from surrounding tissues. The research community was extensively involved through an online survey that incorporated comprehensive protocol details, feasibility assessments, tutorial videos, and illustrative segmentations. Through this collaborative process, consensus emerged to exclude subfield labeling in the hippocampal tail due to limited visibility of internal landmarks and substantial anatomical variability in this region. All proposed boundary guidelines were deemed clear and agreed upon via a Delphi procedure. The harmonized tail protocol has high intra-(Averaged ICC(2,1) > 0.98; Averaged Dice Similarity Coefficient = 0.92) and inter-rater reliability (Averaged ICC(2,k) > 0.98; Averaged Dice Similarity Coefficient = 0.86) and offers a practical framework for replicable segmentation. By establishing standardized guidelines, this protocol enhances comparability of findings across developmental, aging, and clinical research and is compatible with ongoing technological advances.
]]></description>
<dc:creator>de Flores, R.</dc:creator>
<dc:creator>Canada, K. L.</dc:creator>
<dc:creator>Brown, T.</dc:creator>
<dc:creator>Gervais, N. J.</dc:creator>
<dc:creator>Maass, A.</dc:creator>
<dc:creator>Radman, G.</dc:creator>
<dc:creator>Shine, J.</dc:creator>
<dc:creator>Tucker, H. L.</dc:creator>
<dc:creator>Molloy, E. N.</dc:creator>
<dc:creator>Adams, J. N.</dc:creator>
<dc:creator>Reinke, M. B.</dc:creator>
<dc:creator>Bakker, A.</dc:creator>
<dc:creator>Berron, D.</dc:creator>
<dc:creator>Dalton, M. A.</dc:creator>
<dc:creator>Kennedy, K. M.</dc:creator>
<dc:creator>La Joie, R.</dc:creator>
<dc:creator>Mueller, S. G.</dc:creator>
<dc:creator>Ofen, N.</dc:creator>
<dc:creator>Olsen, R. K.</dc:creator>
<dc:creator>Raz, N.</dc:creator>
<dc:creator>Riggins, T.</dc:creator>
<dc:creator>Rodrigue, K. M.</dc:creator>
<dc:creator>Stark, C.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Wisse, L. E.</dc:creator>
<dc:creator>Yushkevich, P. A.</dc:creator>
<dc:creator>Carr, V. A.</dc:creator>
<dc:creator>Daugherty, A. M.</dc:creator>
<dc:creator>Alzheimer's Disease Neuroimaging Initiative,</dc:creator>
<dc:creator>Hippocampal Subfields Group,</dc:creator>
<dc:date>2025-11-20</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689476</dc:identifier>
<dc:title><![CDATA[Harmonized Protocol for Segmentation of the Hippocampal Tail on High-Resolution in vivo MRI from the Hippocampal Subfields Group (HSG)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.20.689516v1?rss=1">
<title>
<![CDATA[
Analyses of bent spindles reveal the mechanics of anaphase B in fission yeast 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689516v1?rss=1</link>
<description><![CDATA[
The mitotic spindle in the fission yeast Schizosaccharomyces pombe is a single bundle of microtubules which elongates to segregate the chromosomes during anaphase B. The mechanical properties of the spindle and the forces driving its elongation remain poorly defined. Here, we analyzed how spindles react to mechanical and genetic perturbations to uncover their mechanical properties. Treatment of cells with osmotic oscillations and blue light led to a consistent phenotype of spindle buckling and breakage in mid-anaphase. The stalling of pole separation and reduced rates of spindle elongation indicated that spindles elongate and buckle under increased mechanical load. The structural integrity of the bent spindles was dependent on Ase1 (PRC1), while the spindle elongation rate was dependent on motor proteins Klp9 (kinesin-6) and Cut7 (kinesin-5). Modeling of bent spindle shapes revealed that most spindles behave mechanically as a beam with a two-fold increase in rigidity in the midzone. Upon reaching a threshold size, bent spindles broke at a specific fragile site near the edge of the spindle midzone. Our findings in this simple fission yeast spindle are relevant to the mechanics of more complex metazoan spindles.

Significance statementO_LIThe anaphase B spindle in S. pombe consists of a microtubule bundle that elongates to move the chromosomes apart. The various forces and mechanical properties of the spindle remain poorly quantified.
C_LIO_LIThe authors establish a method to induce spindle buckling in mid-anaphase. Time-lapse imaging shows that these spindles elongate at reduced rates, buckle as a non-homogeneous beam under mechanical load, and break at a fragile site adjacent to the midzone.
C_LIO_LIThese results provide quantitative and molecular insights into spindle force regulation and structural integrity that are relevant to mitosis in other cell types.
C_LI
]]></description>
<dc:creator>Real-Calderon, P.</dc:creator>
<dc:creator>Fai, T.</dc:creator>
<dc:creator>Daga, R.</dc:creator>
<dc:creator>Lemiere, J.</dc:creator>
<dc:creator>Chang, F.</dc:creator>
<dc:date>2025-11-20</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689516</dc:identifier>
<dc:title><![CDATA[Analyses of bent spindles reveal the mechanics of anaphase B in fission yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.20.689355v1?rss=1">
<title>
<![CDATA[
Congenital CMV infection drives oligoclonal expansion of cytotoxic γδ T cells from early fetal progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689355v1?rss=1</link>
<description><![CDATA[
Gamma delta T cells ({gamma}{delta} T cells) emerge early during human gestation and are uniquely equipped to protect the fetus and infant following in utero infection with viruses such as cytomegalovirus (CMV). Previous research showed that fetal {gamma}{delta} T cells in early gestation are transcriptionally pre-programmed for effector functions in the thymus. Infants with congenital CMV infection (cCMV) exhibit expansions of {gamma}{delta} T cells with CMV-reactive TCRs; however, the functional and transcriptional programming of these innate-like effector cells has not been characterized. Here, we analyzed cord blood mononuclear cells from cCMV+ and uninfected neonates in Uganda using flow cytometry and single-cell RNA and TCR sequencing. We find that {gamma}{delta} T cells in cCMV+ neonates are more differentiated, activated, cytotoxic, and proliferative. TCR repertoires of cCMV+ infants exhibit oligoclonal expansions with an enrichment of {gamma}{delta}TCRs that possess shorter CDR3 lengths and fewer N additions, which suggest they arise from early fetal progenitor cells. These expanded {gamma}{delta} T cell clonotypes in cCMV+ infants are more frequently public and exhibit cytotoxic transcriptional programming. These findings demonstrate that cCMV infection drives an oligoclonal expansion of highly cytotoxic effector {gamma}{delta} T cells with fetal-like TCR features, underscoring their specialized roles in early-life immunity.
]]></description>
<dc:creator>Levan, J.</dc:creator>
<dc:creator>Vaaben, A. V.</dc:creator>
<dc:creator>Tsui, J.</dc:creator>
<dc:creator>Davidson, B. R.</dc:creator>
<dc:creator>Olive, M. E.</dc:creator>
<dc:creator>Callaway, P. C.</dc:creator>
<dc:creator>Nguyen, C. B. T.</dc:creator>
<dc:creator>Ilala, M.</dc:creator>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Hunter, R. D.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Nankya, F.</dc:creator>
<dc:creator>Dorsey, G.</dc:creator>
<dc:creator>Kakuru, A.</dc:creator>
<dc:creator>Muhindo, M.</dc:creator>
<dc:creator>Feeney, M. E.</dc:creator>
<dc:date>2025-11-21</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689355</dc:identifier>
<dc:title><![CDATA[Congenital CMV infection drives oligoclonal expansion of cytotoxic γδ T cells from early fetal progenitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.20.689403v1?rss=1">
<title>
<![CDATA[
"Awe-scillations": EEG spectral and complexity representations of awe 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689403v1?rss=1</link>
<description><![CDATA[
Awe is a positive emotion often accompanied by sensations of vastness and unity, with known benefits for well-being and social behavior. However, its neural underpinnings remain poorly understood. We recorded EEG and autonomic physiology in 23 healthy older participants while they watched a nature-based audiovisual film, The Nature Journey, and subjective ratings of awe were recorded. Awe was associated with decreased alpha and theta spectral power, and increased Lempel Ziv complexity (LZC) - indicating heightened neural signal entropy and increased information processing. These effects partially converged across datasets and awe induction methods, supporting their generalizability. Notably, awe-related increases in LZC correlated with reduced sympathetic activity and greater self-reported awe intensity, but not self-reported joy, suggesting some level of specificity. These results suggest that awe evokes distinctive neurophysiological states linked to both autonomic changes and subjective experience.
]]></description>
<dc:creator>Chen, J. C. C.</dc:creator>
<dc:creator>Mace, G.</dc:creator>
<dc:creator>Ostrand, A.</dc:creator>
<dc:creator>Valtierra, C.</dc:creator>
<dc:creator>Griffith, S.</dc:creator>
<dc:creator>Campusano, R.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Vinson, E.</dc:creator>
<dc:creator>Schlomberg, J.</dc:creator>
<dc:creator>Eshel, M.</dc:creator>
<dc:creator>Suntay, J. M.</dc:creator>
<dc:creator>Rahim, S.</dc:creator>
<dc:creator>Anguera, R.</dc:creator>
<dc:creator>Brown, D. A.</dc:creator>
<dc:creator>Swann, N. C.</dc:creator>
<dc:creator>Mazzucato, L.</dc:creator>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Timmermann, C.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Zanto, T.</dc:creator>
<dc:creator>Ziegler, D.</dc:creator>
<dc:creator>Gazzaley, A.</dc:creator>
<dc:creator>Pasquini, L.</dc:creator>
<dc:date>2025-11-21</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689403</dc:identifier>
<dc:title><![CDATA["Awe-scillations": EEG spectral and complexity representations of awe]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.20.689520v1?rss=1">
<title>
<![CDATA[
Drug-induced phospholipidosis as an artifact in antiviral drug repurposing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689520v1?rss=1</link>
<description><![CDATA[
Drug repurposing in principle can speed antiviral drug discovery. Among the molecules most frequently advanced in such repurposing efforts are a group of structurally diverse cationic amphiphilic drugs (CADs). While CADs have shown micromolar to mid-nanomolar antiviral activity in cell-based assays, they can induce phospholipidosis, confounding in COVID-19 repurposing. A barrier to the identification of phospholipidosis inducers has been the involved nature of the microscopy assays used to characterize them. To ease the identification of these artifacts, we describe a rapid microplate-based assay to detect phospholipidosis. Leveraging this assay, we quantified the prevalence of phospholipidosis-inducers across several cell-based antiviral repurposing screens. We selected 40 drugs reported to have micromolar antiviral activities and found that 26 of them (65%) induced phospholipidosis within the same concentration range as their reported antiviral activities. Intriguingly, we identified four non-CADs that also induce phospholipidosis, revealing a new group of drugs that can lead to this toxic event and highlighting the importance of facile experimental assays to detect it. Understanding how phospholipidosis can confound antiviral drug discovery, and its rapid detection, will help prevent what is an apparently general artifact, active across viruses, from distracting investigators from potentially more useful candidates.
]]></description>
<dc:creator>Glenn, I. S.</dc:creator>
<dc:creator>Paris, L.</dc:creator>
<dc:creator>White, A. D.</dc:creator>
<dc:creator>Garda, V. G.</dc:creator>
<dc:creator>Montano, M.</dc:creator>
<dc:creator>Khalid, M. M.</dc:creator>
<dc:creator>Kao, A. W.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2025-11-21</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689520</dc:identifier>
<dc:title><![CDATA[Drug-induced phospholipidosis as an artifact in antiviral drug repurposing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.20.689596v1?rss=1">
<title>
<![CDATA[
Saxiphilin functions as a "toxin sponge" protein that counteracts the effects of saxitoxin poisoning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689596v1?rss=1</link>
<description><![CDATA[
Saxitoxin (STX) is among the most potent toxins known, is classified as a chemical weapon, and is the archetype of the paralytic shellfish toxin (PST) family produced by marine and freshwater harmful algal blooms (HAB)1-3. STX causes paralysis and death through inhibition of voltage-gated sodium channels (NaVs), has no antidote, and poses a public health and commercial fishing threat due to its accumulation in seafood and increasing HAB occurrences1,4. Although STX is lethal to varied vertebrates5-7, including humans4,8,9, certain frogs resist STX poisoning6,7. This phenotype is thought to depend on soluble  toxin sponge high-affinity STX binding proteins (saxiphilins, Sxphs)6,7,10,11 that provide resistance through a competition mechanism different from classic  target site ion channel mutation mechanisms12-14. Here, we test this idea directly and show that a single dose of American bullfrog (Rana catesbeiana) Sxph (RcSxph) is sufficient to counteract STX neurotoxicity and lethality in mice using paradigms where STX and Sxph are administered together or sequentially in either order. Importantly, this function requires the RcSxph high-affinity STX binding site. Our findings provide in vivo validation that Sxphs are  toxin sponges that protect against STX poisoning and highlight the potential to harness this protein class as antidotes for PSTs and other toxins.
]]></description>
<dc:creator>Nixon, S. A.</dc:creator>
<dc:creator>Zakrzewska, S.</dc:creator>
<dc:creator>Jang, S.</dc:creator>
<dc:creator>Huang, K.</dc:creator>
<dc:creator>Barra, A.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Goss, D. R.</dc:creator>
<dc:creator>Park, E.</dc:creator>
<dc:creator>Du Bois, J.</dc:creator>
<dc:creator>Minor, D. L.</dc:creator>
<dc:date>2025-11-21</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689596</dc:identifier>
<dc:title><![CDATA[Saxiphilin functions as a "toxin sponge" protein that counteracts the effects of saxitoxin poisoning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.21.689519v1?rss=1">
<title>
<![CDATA[
Increased mannosylation of extracellular vesicles in Long COVID plasma provides a potential therapeutic target for Galanthus nivalis agglutinin (GNA) affinity resin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689519v1?rss=1</link>
<description><![CDATA[
There is no proven therapy for Long COVID, a post-acute illness characterized by a myriad of diverse symptoms including fatigue, dyspnea, and brain fog following SARS-CoV-2 infection. Extracellular vesicles (EVs) have been implicated in Long COVID pathogenesis by promoting viral and inflammatory signaling with their molecular cargo. In this study, we investigated whether EV abundance and glycome characteristics are altered in plasma from people with Long COVID and whether they can be targeted for removal using a glycan-binding affinity resin. Large (100-500 nm) and small (40-200 nm) EVs were isolated from plasma of participants in the post-acute phase of COVID-19 and analyzed by nanoparticle flow cytometry to measure concentration and glycan characteristics. Plasma of those with Long COVID contained elevated levels of both large and small EVs, and mannose-positive large EVs were significantly increased in comparison to recovered controls (p < 0.05). EV capture assays using Galanthus nivalis agglutinin (GNA) affinity resin demonstrated small EV removal positively correlated with mannose-positive EV abundance (r = 0.341, p < 0.05). NanoString analyses identified seven EV-associated miRNAs significantly depleted by GNA affinity resin treatment of plasma. PROGENy pathway inference of validated miRNA-mRNA interactions suggests these reductions may lead to a downregulation of JAK-STAT signaling and upregulation of Estrogen, VEGF, and PI3K pathways, resulting in a favorable rebalancing of immune and tissue-repair networks. These findings reveal specific glycome EV-miRNA cargo signatures in Long COVID and the potential clinical benefits of a lectin capture therapeutic strategy to remove these pathogenic vesicles and their inflammatory cargo.
]]></description>
<dc:creator>Pesqueira Sanchez, M. A.</dc:creator>
<dc:creator>de Necochea Campion, R.</dc:creator>
<dc:creator>Dalhuisen, T.</dc:creator>
<dc:creator>Fehrman, E. A.</dc:creator>
<dc:creator>Chhabra, P. S.</dc:creator>
<dc:creator>Kelly, J. D.</dc:creator>
<dc:creator>Martin, J. N.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Henrich, T. J.</dc:creator>
<dc:creator>Peluso, M. J.</dc:creator>
<dc:creator>LaRosa, S. P.</dc:creator>
<dc:date>2025-11-21</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689519</dc:identifier>
<dc:title><![CDATA[Increased mannosylation of extracellular vesicles in Long COVID plasma provides a potential therapeutic target for Galanthus nivalis agglutinin (GNA) affinity resin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.20.689570v1?rss=1">
<title>
<![CDATA[
High extraversion enhances attentional control through dynamic network reorganization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689570v1?rss=1</link>
<description><![CDATA[
The extraversion-introversion dimension of personality is hypothesized to differ based on low or high cortical arousal, respectively. Notably, high cortical arousal in introverts is thought to underlie increased distractibility. Here, we assess fMRI while participants meditate (focused attention to their breath) under three levels of auditory distraction: no, low and high. Whereas introverts exhibited worsening attentional focus on their breath with increasing distraction, extraverts retained their ability to focus attention despite distraction. Dynamic functional connectivity analysis indicated that extraverts exhibited less globally efficient and less modular networks, which may prevent distracting stimuli from creating interference. Furthermore, connectivity strengths amongst the default mode, central executive, and salience networks were increased for extraverts and decreased in introverts during high focused attention; potentially indicating distinct cognitive processes that support attentional control. These findings support the hypothesis regarding differing levels of cortical arousal in extraverts and introverts and extend personality theory by linking the extraversion dimension to attentional control and functional connectivity dynamics.
]]></description>
<dc:creator>Chen, J. C. C.</dc:creator>
<dc:creator>Nandi, B.</dc:creator>
<dc:creator>Campusano, R.</dc:creator>
<dc:creator>Jiang, F.</dc:creator>
<dc:creator>Ziegler, D.</dc:creator>
<dc:creator>Gazzaley, A.</dc:creator>
<dc:creator>Zanto, T.</dc:creator>
<dc:date>2025-11-22</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689570</dc:identifier>
<dc:title><![CDATA[High extraversion enhances attentional control through dynamic network reorganization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.21.689741v1?rss=1">
<title>
<![CDATA[
Genetic variation of human G6PD impacts Red Blood Cell transfusion efficacy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689741v1?rss=1</link>
<description><![CDATA[
Glucose-6-phosphate dehydrogenase (G6PD) deficiency, the most common human enzymopathy, affects 6% of the global population, yet its impact on blood storage and transfusion efficacy remains undefined. We integrated genome-metabolome-proteome analyses of 13,091 blood donors (362 G6PD SNPs), validated in a recalled cohort (n=643), linked donor-recipient databases, humanized mouse models (canonical, African A- [V68M+N126D], Mediterranean [S188F]), and a prospective sickle cell disease study. Common G6PD variants reduced protein abundance, reprogrammed redox metabolism, and increased storage hemolysis. In mice, G6PD-deficient RBCs showed lower post-transfusion recovery, higher oxidative stress, and impaired renal oxygenation. Clinically, recipients of G6PD-deficient units exhibited smaller hemoglobin increments and reduced RBC L{superscript 1}Cr-survival (-8% at 24 h; -12% at 4 weeks). Structural studies revealed kinetic fragility for A- and thermodynamic fragility for Med-, linking genotype to protein instability and transfusion outcome. These findings identify donor G6PD genotype as a determinant of transfusion efficacy, supporting genotype-aware inventory-management strategies.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/689741v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Karafin, M. S.</dc:creator>
<dc:creator>Issaian, A. V.</dc:creator>
<dc:creator>Bevers, S.</dc:creator>
<dc:creator>Reisz, J. A.</dc:creator>
<dc:creator>Hay, A.</dc:creator>
<dc:creator>Keele, G. R.</dc:creator>
<dc:creator>Dzieciatkowska, M.</dc:creator>
<dc:creator>Cendali, F. I.</dc:creator>
<dc:creator>Haiman, Z. B.</dc:creator>
<dc:creator>Key, A. M.</dc:creator>
<dc:creator>Nemkov, T.</dc:creator>
<dc:creator>Stephenson, D.</dc:creator>
<dc:creator>Marsh, E.</dc:creator>
<dc:creator>Moore, A.</dc:creator>
<dc:creator>Palha, M. S.</dc:creator>
<dc:creator>Legenzov, E. A.</dc:creator>
<dc:creator>Lamb, D. R.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>Kleinman, S.</dc:creator>
<dc:creator>Norris, P. J.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>Vallese, F.</dc:creator>
<dc:creator>Palsson, B. O.</dc:creator>
<dc:creator>Spitalnik, S.</dc:creator>
<dc:creator>Kao, J. P.</dc:creator>
<dc:creator>Roubinian, N. H.</dc:creator>
<dc:creator>Janetzko, J.</dc:creator>
<dc:creator>Page, G. P.</dc:creator>
<dc:creator>Eisenmesser, E. Z.</dc:creator>
<dc:creator>Zimring, J. C.</dc:creator>
<dc:creator>Buehler, P. W.</dc:creator>
<dc:creator>D'Alessandro, A.</dc:creator>
<dc:date>2025-11-22</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689741</dc:identifier>
<dc:title><![CDATA[Genetic variation of human G6PD impacts Red Blood Cell transfusion efficacy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.21.689510v1?rss=1">
<title>
<![CDATA[
Sphingosine-1-phosphate receptor modulators resensitize FLT3-ITD acute myeloid leukemia cells with NRAS mutations to FLT3 inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689510v1?rss=1</link>
<description><![CDATA[
FLT3 inhibitor efficacy in AML with FLT3-ITD is short-lived, frequently due to new mutations, most commonly in NRAS. Sphingosine kinase 1 (SPHK1), which phosphorylates sphingosine to generate sphingosine-1-phosphate (S1P), is upregulated and localized to the plasma membrane in RAS-mutated cells. We studied S1P and FLT3 co-targeting to overcome FLT3 inhibitor resistance in NRAS-mutated FLT3-ITD AML cells. NRAS-mutated FLT3-ITD AML cell lines and patient blasts were treated with FLT3 inhibitors and/or S1P receptor (S1PR) modulators. FLT3 inhibitor sensitivity was assessed by immunoblotting, cytotoxicity and apoptosis assays. Co-treatment was also assessed in vivo in an orthotopic mouse model. Downstream RAS and SPHK1 effectors were measured by immunoblotting and qRT-PCR. The S1PR modulators fingolimod (FTY720) and mocravimod (KRP-203) resensitized FLT3-ITD-expressing MOLM-14 and MV4-11 human AML cells with G12D, G12S, Q61K or Q61H, but not G12C, and patient blasts with G13D or G13V NRAS mutations to FLT3 inhibitors. Moreover, FTY720 co-treatment resensitized G12D NRAS-mutated M14(R)701 cells to gilteritinib in vivo. Co-treatment inactivated ERK, transcriptionally downregulated SPHK1, and inactivated downstream AKT, p70S6K and BAD, with inactivation abrogated by constitutive SPHK1 expression. The clinically applicable S1PR modulators fingolimod and mocravimod resensitize NRAS-mutated FLT3-ITD AML cells to FLT3 inhibitors, supporting potential clinical efficacy of these combinations.
]]></description>
<dc:creator>Baer, M. R.</dc:creator>
<dc:creator>Chatterjee, A.</dc:creator>
<dc:creator>Ali, M. K. M.</dc:creator>
<dc:creator>Bailey, C. M.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Small, D.</dc:creator>
<dc:creator>Smith, C. C.</dc:creator>
<dc:creator>Traer, E. C.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Silvestri, G.</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689510</dc:identifier>
<dc:title><![CDATA[Sphingosine-1-phosphate receptor modulators resensitize FLT3-ITD acute myeloid leukemia cells with NRAS mutations to FLT3 inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.23.690004v1?rss=1">
<title>
<![CDATA[
Bacteriophage genome-wide transposon mutagenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.23.690004v1?rss=1</link>
<description><![CDATA[
Bacteriophages have genomes that span a wide size range, are densely packed with coding sequences, and frequently encode genes of unknown function. Classical forward genetics has defined essential genes for phage replication in a few model systems but remains laborious and non-scalable. Unbiased functional genomics approaches are therefore needed for phages, particularly for large lytic phages. Here, we develop a phage transposon sequencing (TnSeq) platform that uses the mariner transposase to insert an anti-CRISPR selectable marker into phage genomes. CRISPR-Cas13a-based enrichment of transposed phages followed by pooled sequencing identifies both fitness-conferring and dispensable genes. Using the Pseudomonas aeruginosa-infecting nucleus-forming jumbo phage {Phi}KZ (280,334 bp; 371 predicted genes) as a model, we show that [~]110 genes are fitness-conferring via phage TnSeq. These include conserved essential genes involved in phage nucleus formation, protein trafficking, transcription, DNA replication, and virion assembly. We also isolate hundreds of individual phages with insertions in non-essential genes and reveal conditionally essential genes that are specifically required in clinical isolates, at environmental temperature, or in the presence of a defensive nuclease. Phage TnSeq is a facile, scalable technology that can define essential phage genes and generate knockouts in all non-essential genes in a single experiment, enabling conditional genetic screens in phages and providing a broadly applicable resource for phage functional genomics.
]]></description>
<dc:creator>Chan, A. W.</dc:creator>
<dc:creator>Yee, W.-X.</dc:creator>
<dc:creator>Mozumdar, D.</dc:creator>
<dc:creator>Kokontis, C. Y.</dc:creator>
<dc:creator>Rojas-Montero, M.</dc:creator>
<dc:creator>Yuping, L.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.23.690004</dc:identifier>
<dc:title><![CDATA[Bacteriophage genome-wide transposon mutagenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.24.690213v1?rss=1">
<title>
<![CDATA[
A geothermal amoeba sets a new upper temperature limit for eukaryotes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.24.690213v1?rss=1</link>
<description><![CDATA[
The study of temperature limits has transformed our knowledge of the boundaries of life but has been largely focused on bacteria and archaea. We isolated a novel geothermal amoeba, Incendiamoeba cascadensis, that divides at 63{degrees}C (145.4{degrees}F), establishing a new record for the upper temperature limit across all eukaryotes. We demonstrated cellular proliferation with growth experiments and visualized mitosis via expansion microscopy. Using high-temperature live-cell imaging, we quantified movement up to 64{degrees}C. We assembled the genome of I. cascadensis and using comparative genomics found an enrichment of genes related to proteostasis, genome stability, and sensing the external environment. Taken together, our findings challenge the current paradigm of temperature constraints on eukaryotic cells and reshape our understanding of where and how eukaryotic life can persist.
]]></description>
<dc:creator>Rappaport, H. B.</dc:creator>
<dc:creator>Petek-Seoane, N. A.</dc:creator>
<dc:creator>Tyml, T.</dc:creator>
<dc:creator>Mikus, F.</dc:creator>
<dc:creator>Labutti, K.</dc:creator>
<dc:creator>Ani, G.</dc:creator>
<dc:creator>Niblo, J. K.</dc:creator>
<dc:creator>MacVicar, E.</dc:creator>
<dc:creator>Shepherd, R. M.</dc:creator>
<dc:creator>de la Higuera, I.</dc:creator>
<dc:creator>Lord, S. J.</dc:creator>
<dc:creator>Dey, G.</dc:creator>
<dc:creator>Wolfe, G. V.</dc:creator>
<dc:creator>Dudin, O.</dc:creator>
<dc:creator>Sukenik, S.</dc:creator>
<dc:creator>Katz, L. A.</dc:creator>
<dc:creator>Stedman, K. M.</dc:creator>
<dc:creator>Skruber, K.</dc:creator>
<dc:creator>Schulz, F.</dc:creator>
<dc:creator>Mullins, R. D.</dc:creator>
<dc:creator>Oliverio, A. M.</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.24.690213</dc:identifier>
<dc:title><![CDATA[A geothermal amoeba sets a new upper temperature limit for eukaryotes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.21.688964v1?rss=1">
<title>
<![CDATA[
Steady-state epithelial apical flatness is characterized by MLCK morphodynamics and asynchronous Ca2+ oscillations, but not underlying ECM geometry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.688964v1?rss=1</link>
<description><![CDATA[
The canonical simple epithelium is a flat sheet-like tissue of horizontally packed cells. While the basal surface is delineated by the basement membrane of extracellular matrix (ECM), little is known about how a flat apical surface is maintained, or if apical/basal dynamics are coordinated. The current study tests the role of the apical domain, to define mechanisms involved in maintaining a flat apical geometry in an epithelium. When the basal geometry is modulated, Madin-Darby Canine Kidney (MDCK) cells adjust their morphology to maintain an overall apical flatness of the confluent layer. Pharmacological and transgenic disruption of non-muscle myosin ATPase, and MLCK activity results in an uneven apical structure, and overall loss of the flat geometry typical of a confluent epithelium. Surprisingly, transgenic experimentation showed that forces maintaining individual MDCK cell flatness are cell-autonomous. Finally, Ca2+ imaging reveals an asynchronous calcium flux across a confluent epithelium, suggesting a myosin II-mediated mechanism for maintaining a flat apical architecture. Our results highlight that apical/basal cellular surfaces may not be tightly coordinated, but rather independently regulated. This study provides a new paradigm for how apical flatness is regulated at steady state.

Impact StatementO_LIHow surface flatness of an epithelium is established and maintained is unknown; the data reveal that apical flatness is controlled independently from basement membrane geometry, and require balancing of myosin II morphodynamics.
C_LI
]]></description>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Herrmann, E. G.</dc:creator>
<dc:creator>Hyer, J. M.</dc:creator>
<dc:creator>Hua, L. L.</dc:creator>
<dc:creator>Mikawa, T.</dc:creator>
<dc:date>2025-11-25</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.688964</dc:identifier>
<dc:title><![CDATA[Steady-state epithelial apical flatness is characterized by MLCK morphodynamics and asynchronous Ca2+ oscillations, but not underlying ECM geometry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.21.689836v1?rss=1">
<title>
<![CDATA[
Modifications to the gut microbiome alter bone matrix proteomics and fracture toughness at the cellular scale 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689836v1?rss=1</link>
<description><![CDATA[
The gut microbiome can regulate the strength of bone matrix but the specific changes in matrix function and composition are not yet understood. Here we introduce micropillar splitting to determine the fracture toughness of matrix at the cellular scale in concert with proteomic analysis of bone matrix when the gut microbiome was altered by oral antibiotics (ampicillin+neomycin). Male mice were divided into four groups (n = 3-4/group): (1) Unaltered (no alteration), (2) Continuous (alteration from 4-24 weeks), (3) Delayed (alteration from 16-24 weeks) and (4) Reconstituted (alteration from 1-16 weeks following by reconstitution). Micropillars (5 {micro}m diameter) were fabricated using focused ion beam milling on femur cross-sections in regions of matrix formed either before or after changes in the microbiome (16 weeks) (n = 4/group). Proteomics was used to identify differences in matrix protein composition. Micropillar fracture toughness differed by group (p < 0.001) and region (p < 0.001). Fracture toughness in the Unaltered group (1.34 {+/-} 0.32 MPa{surd}m, mean {+/-} SD) was substantially greater than the Continuous group (0.95 {+/-} 0.20) and the Delayed group (0.90 {+/-} 0.21) but not different from the Reconstituted group (1.22 {+/-} 0.25). Bone matrix formed from 16-24 weeks of age had lower fracture toughness than matrix formed before 16 weeks of age in all groups. Notably, micropillar splitting was substantially more precise than whole bone testing; whole bone notched 3-point bending tests did not detect differences in fracture toughness. Proteomics identified 46 extracellular matrix proteins that were differentially abundant between groups, including decreased abundance of Periostin (q < 0.001) and Emilin-1 (q < 0.001) in groups with impaired bone matrix. These findings demonstrate that modifications to the gut microbiome lead to changes in bone matrix throughout cortical bone volume and establish micropillar splitting as a high-precision approach for characterizing matrix material properties.

Plain Language SummaryThis study investigated the effect of the gut microbiome on the brittleness of bone as a material. We used a new technique for measuring the brittleness of bone that involves making microscopic pillars on the bone surface and splitting them down the middle to measure a material property called fracture toughness. Mice with altered gut microbiomes had bone with reduced fracture toughness (by 35-40%). Restoration of an altered gut microbiome for two months reversed the effects. The effect of the microbiome on bone occurred throughout the whole bone and was not limited to regions of the bone formed after a change in the gut microbiome.
]]></description>
<dc:creator>Hunt, N. K.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:creator>Stephen, S. J.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Schurman, C. A.</dc:creator>
<dc:creator>King, C. D.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Vashishth, D.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:creator>Hassani, M.</dc:creator>
<dc:creator>Hernandez, C. J.</dc:creator>
<dc:date>2025-11-25</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689836</dc:identifier>
<dc:title><![CDATA[Modifications to the gut microbiome alter bone matrix proteomics and fracture toughness at the cellular scale]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.22.689595v1?rss=1">
<title>
<![CDATA[
Injury-induced tau pathology promotes aggressive behavior in Drosophila without neurodegeneration. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.22.689595v1?rss=1</link>
<description><![CDATA[
The microtubule-associated protein tau is implicated in neurodegenerative diseases, but its physiological roles remain poorly understood. Here, we find that pan-neuronal expression of human tau (HsTau) in Drosophila coupled with injury triggers hyper-aggression in male flies, which is absent in flies expressing non-phosphorylatable tau. These behavioral manifestations result from activation of dopaminergic circuits without neurodegeneration. Using in vitro reconstitution assays, we find that phosphorylated HsTau maintains microtubule binding but loses its ability to suppress catastrophes, thereby promoting microtubule dynamicity. In contrast, unphosphorylated HsTau and fly tau (DmTau) stabilize microtubules by reducing catastrophe frequency. Our findings challenge the canonical view of tau as a simple microtubule stabilizer and instead position it as a dynamic regulator of microtubule function and neuronal excitability. These results reveal how acute tau phosphorylation can alter neural circuit function and behavior prior to neurodegeneration, providing new insights into taus physiological and pathological roles.
]]></description>
<dc:creator>Maxson, R.</dc:creator>
<dc:creator>Smoyer, C. J.</dc:creator>
<dc:creator>Hampton, M. F.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Wiese, K.</dc:creator>
<dc:creator>Yee, C.</dc:creator>
<dc:creator>Singh, A.</dc:creator>
<dc:creator>Funtila, A.</dc:creator>
<dc:creator>McKenney, R. J.</dc:creator>
<dc:creator>Ori-McKenney, K. M.</dc:creator>
<dc:date>2025-11-25</dc:date>
<dc:identifier>doi:10.1101/2025.11.22.689595</dc:identifier>
<dc:title><![CDATA[Injury-induced tau pathology promotes aggressive behavior in Drosophila without neurodegeneration.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.22.689843v1?rss=1">
<title>
<![CDATA[
Immunological Differences in Atopic Dermatitis Across Age Groups: Insights from Single-Cell Multi-Omics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.22.689843v1?rss=1</link>
<description><![CDATA[
BackgroundAtopic dermatitis (AD) occurs across all ages but presents distinct clinical and immunologic features between children, adults, and older adults. The molecular programs underlying these age-specific immune differences remain poorly understood.

MethodsWe performed single-cell multi-omics profiling of peripheral blood mononuclear cells (PBMCs) from 29 AD patients and 29 matched healthy controls (HC), spanning pediatric (0-17 years), adult (18-59 years), and geriatric ([&ge;]60 years) groups. Using Cellular Indexing of Transcriptomes and Epitopes by sequencing (CITE-seq), we simultaneously quantified transcriptomic (RNA) and surface proteomic (ADT) profiles across [~]280,000 immune cells. Integrated analyses identified 30 immune subsets for cell-type proportion and differential expression analyses. Machine-learning classifiers were trained on significant gene and protein features to distinguish AD subgroups by age.

ResultsCompared with HC, AD blood showed enrichment of CD14+ monocytes, plasmacytoid dendritic cells, and CD4+ proliferating T cells. Within AD, pediatric patients had increased {gamma}{delta} T cells, naive CD4+, and naive CD8+ T cells, while geriatric patients exhibited more CD4+ cytotoxic and CD8+ central memory T cells, indicating a shift from naive to effector predominance with aging. Transcriptomic and proteomic analyses revealed distinct programs: pediatric AD was enriched for IL-10 and cytokine-cytokine receptor signaling; adult AD demonstrated activation of metabolic and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-{kappa}B)/Th1/Th17 pathways; and geriatric AD exhibited reduced adaptive immune activity but increased innate signaling. Machine-learning models based on differentially expressed genes and proteins accurately classified AD age groups (transcript-based F1 = 0.70, AUC = 0.79), identifying stable markers such as IRF2, PDK4, ZFP90, CD21, CD94, and CD122.

ConclusionsSingle-cell multi-omics profiling revealed immune differences across the AD lifespan, transitioning from developmental tolerance in children to inflammatory and metabolic activation in adults to enhanced innate signaling in geriatric individuals. These findings highlight molecular signatures that could support age-specific diagnostics and therapeutic strategies for AD across the lifespan.
]]></description>
<dc:creator>Baldonado, G. C. L.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Jin, J.</dc:creator>
<dc:creator>Fang, X.</dc:creator>
<dc:creator>Ildardashty, A.</dc:creator>
<dc:creator>Braun, M.</dc:creator>
<dc:creator>Neuhaus, I. M.</dc:creator>
<dc:creator>Mathes, E.</dc:creator>
<dc:creator>Bhutani, T.</dc:creator>
<dc:creator>Liao, W.</dc:creator>
<dc:date>2025-11-25</dc:date>
<dc:identifier>doi:10.1101/2025.11.22.689843</dc:identifier>
<dc:title><![CDATA[Immunological Differences in Atopic Dermatitis Across Age Groups: Insights from Single-Cell Multi-Omics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.25.690589v1?rss=1">
<title>
<![CDATA[
Crystallographic Ensembles Reveal the Structural Basis of Binding Entropy in SARS-CoV2 Macrodomain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.25.690589v1?rss=1</link>
<description><![CDATA[
Structure-based drug design has traditionally focused on optimizing static, enthalpic interactions between ligands and proteins or on displacing binding site solvent molecules to entropically favor binding. A potentially large contributor to binding thermodynamics is the difference in conformational entropy of the protein upon binding a ligand; however, this has been difficult to quantify especially in high throughput. Here, through multiconformer ensemble modeling of hundreds of ligand-bound SARS-CoV-2 Macrodomain (Mac1) X-ray structures, we show how ligand binding reorganizes both protein conformational entropy and water molecules. By applying an optimal transport-based clustering algorithm, we show how specific protein-ligand interactions patterns drive the magnitude and spatial redistribution of conformational entropy and solvent networks. Using isothermal titration calorimetry (ITC), we demonstrate a correlation between experimental binding thermodynamics and conformational entropy estimated from structural ensemble models, showing that increased conformational heterogeneity and a less connected hydrogen-bonded water network lead to more entropic binding. These results establish a framework for extracting thermodynamically meaningful information from crystallographic ensembles, enabling the integration of entropic effects into prospective, ensemble-aware drug discovery.
]]></description>
<dc:creator>Seo, L.</dc:creator>
<dc:creator>Farran, I.</dc:creator>
<dc:creator>Aslam, A.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Jaishankar, P.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Renslo, A.</dc:creator>
<dc:creator>Wankowicz, S. A.</dc:creator>
<dc:date>2025-11-26</dc:date>
<dc:identifier>doi:10.1101/2025.11.25.690589</dc:identifier>
<dc:title><![CDATA[Crystallographic Ensembles Reveal the Structural Basis of Binding Entropy in SARS-CoV2 Macrodomain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.24.690328v1?rss=1">
<title>
<![CDATA[
Spatial Transcriptomics Reveals CXCL12+ Fibroblasts as Central Immune Organizers through CXCR4 Signaling in Abdominal Aortic Aneurysm 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.24.690328v1?rss=1</link>
<description><![CDATA[
BACKGROUNDAbdominal aortic aneurysm (AAA) is characterized by sterile inflammation, immune cell infiltration, and stromal remodeling that progressively weaken the aortic wall, leading to life-threatening aortic rupture. The molecular mechanisms and spatial organization of immune-stromal interactions in human tissue are poorly understood, limiting the potential to develop effective pharmacological therapy for AAA.

METHODSIn this observational cross-sectional study, formalin-fixed, paraffin-embedded tissues from 11 AAA patients and 12 controls were analyzed by Xenium spatial transcriptomics. Cellular states and localization within tissue architecture were mapped to identify cellular neighborhoods and infer cell-cell communication.

RESULTSWe generated a high-resolution spatial transcriptomics atlas of 581,664 cells in 26 clusters. AAAs showed a significant loss of contractile smooth muscle cells, expansion of pro-angiogenic endothelial subsets, and broad infiltration of immune cells. These inflammatory changes were accompanied by expansion of activated, universal, and CXCL12 adventitial fibroblasts. Spatial transcriptomic analysis revealed fibroblast-immune colocalization and adventitial tertiary lymphoid organs. Inferred signaling pathway analysis identified increased interactions between CXCL12 fibroblasts and CXCR4 T and B cells in the adventitia of AAAs. Fibroblasts that expressed CXCL12 had significantly more immune cell neighbors than fibroblasts that did not, suggesting that they serve as stromal hubs for adaptive immune clustering. Genome-wide association analysis linked AAA heritability to fibroblasts, modulated smooth muscle cells, and foamy macrophages.

CONCLUSIONOur novel high-resolution spatial transcriptomic atlas of human AAAs revealed coordinated pathogenic reprogramming of stromal and immune cells, defined by smooth muscle cell depletion, fibroblast activation, endothelial remodeling, and disproportionate expansion of immune cells. Through CXCR4 signaling, CXCL12 fibroblasts serve as central organizers of immune niches, suggesting stromal-immune crosstalk as a therapeutic target in AAA.

CLINICAL PERSPECTIVESWhat Is New?

O_LIWe generated the first subcellular-resolution spatial transcriptomic atlas of human abdominal aortic aneurysm (AAA), with >580,000 cells identified from aortic tissue sections
C_LIO_LIWe identified CXCL12+ fibroblasts as central stromal hubs that organize adaptive immune niches through CXCR4-mediated crosstalk with B and T cells
C_LIO_LIWe discovered that stromal populations carry the strongest genetic enrichment for AAA risk, notably fibroblast and modulated smooth muscle cell populations
C_LI

What Are The Clinical Implications?

O_LIThese findings position stromal-immune interactions, particularly the CXCL12-CXCR4 axis, as a potential therapeutic target to slow AAA progression
C_LIO_LIThe spatial atlas provides a framework for mechanistic studies and drug-discovery efforts, guiding future interventions aimed at modifying the microenvironment that destabilizes the aneurysmal aortic wall
C_LI
]]></description>
<dc:creator>Levy-Lambert, D.</dc:creator>
<dc:creator>Ramirez, J. L.</dc:creator>
<dc:creator>Shaikh, S.</dc:creator>
<dc:creator>De Jeronimo Diaz, C.</dc:creator>
<dc:creator>Huang, A. G.</dc:creator>
<dc:creator>Combes, A.</dc:creator>
<dc:creator>Fragiadakis, G.</dc:creator>
<dc:creator>Fidler, T.</dc:creator>
<dc:creator>Oskowitz, A.</dc:creator>
<dc:date>2025-11-29</dc:date>
<dc:identifier>doi:10.1101/2025.11.24.690328</dc:identifier>
<dc:title><![CDATA[Spatial Transcriptomics Reveals CXCL12+ Fibroblasts as Central Immune Organizers through CXCR4 Signaling in Abdominal Aortic Aneurysm]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.25.690488v1?rss=1">
<title>
<![CDATA[
Neuronal APOE4 alone is sufficient to drive tau pathology, neurodegeneration, and neuroinflammation in an Alzheimer's disease mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.25.690488v1?rss=1</link>
<description><![CDATA[
Apolipoprotein E4 (APOE4), the strongest genetic risk factor for late-onset Alzheimers disease (AD), exacerbates tau tangles, amyloid plaques, neurodegeneration, and neuroinflammation--the pathological hallmarks of AD. While astrocytes are the primary producers of APOE in the CNS, neurons increase APOE expression under stress and aging. Prior work established that neuronal APOE4 is essential for AD pathogenesis, but whether it is sufficient to drive disease remained unknown. We generated a PS19 tauopathy mouse model selectively expressing APOE4 in neurons. Neuronal APOE4 alone proved sufficient to promote pathological tau accumulation and propagation, neurodegeneration, and neuroinflammation to levels comparable to a tauopathy model with human APOE4 knocked-in globally. Single-nucleus RNA sequencing further revealed similar transcriptomic changes in neurons and glia of both models. Together, these findings demonstrate that neuronal APOE4 alone can initiate and propagate AD pathologies, underscoring its pivotal role in disease pathogenesis and its potential as a therapeutic target.
]]></description>
<dc:creator>Blumenfeld, J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Kim, M. J.</dc:creator>
<dc:creator>Yip, O.</dc:creator>
<dc:creator>Yao, L.</dc:creator>
<dc:creator>De Leon, S.</dc:creator>
<dc:creator>Fulthorpe, R.</dc:creator>
<dc:creator>Shostak, D.</dc:creator>
<dc:creator>Platow, Z.</dc:creator>
<dc:creator>Suan, K.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Koutsodendris, N.</dc:creator>
<dc:creator>Ellis, C.</dc:creator>
<dc:creator>Nguyen, J.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:date>2025-11-29</dc:date>
<dc:identifier>doi:10.1101/2025.11.25.690488</dc:identifier>
<dc:title><![CDATA[Neuronal APOE4 alone is sufficient to drive tau pathology, neurodegeneration, and neuroinflammation in an Alzheimer's disease mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.26.689708v1?rss=1">
<title>
<![CDATA[
HDL-associated phosphatidylserine blunts myeloid activation and protects from atherosclerosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.26.689708v1?rss=1</link>
<description><![CDATA[
Background and aimsLipids play a critical role in atherosclerosis. Low-density lipoprotein (LDL)-cholesterol and certain lipid classes like sphingomyelins are associated with inflammation and poor cardiovascular outcomes. Phosphatidylserine (PS), on the other hand, is a negatively charged anti-inflammatory phospholipid class involved in efferocytosis. In this study, we sought to investigate its anti-atherosclerotic properties through a combination of complementary human lipidomics analyses, in vitro assays and in vivo experiments in Apoe-/- mice.

MethodsHuman lipidomics studies were performed on the 300OB cohort comprising 300 obese and overweight individuals at risk of cardiovascular disease. In vitro assays were carried out using human monocytes and macrophages, and in vivo experiments included histopathological, immunophenotyping and single-cell transcriptomic analyses.

ResultsIn humans, we identified PS as an anti-inflammatory and atheroprotective biomarker. Hence, we developed a high-density lipoprotein (HDL)-like formulation enriched in PS to exploit its properties in a targeted fashion in mice. In vitro, this formulation potently inhibited inflammatory cytokine production on human myeloid cells. Our in-depth in vivo experiments provided evidence of the formulations potent plaque-stabilizing and anti-inflammatory actions. These effects were mediated by a shift in the monocyte/macrophage compartment toward homeostatic/repairing phenotypes.

ConclusionsCollectively, our results demonstrate that HDL-associated PS potently suppresses inflammation and atheroprogression, and holds promise as a viable approach to improve immunomodulatory therapies.
]]></description>
<dc:creator>Bernal, A.</dc:creator>
<dc:creator>Hamid, A.</dc:creator>
<dc:creator>Grao-Roldan, C.</dc:creator>
<dc:creator>Scarpa, A.</dc:creator>
<dc:creator>Sanchez, I.</dc:creator>
<dc:creator>Nicolas-Avila, J. A.</dc:creator>
<dc:creator>Pena-Couso, L.</dc:creator>
<dc:creator>Yaghmour, M. H.</dc:creator>
<dc:creator>Benguria, A.</dc:creator>
<dc:creator>Rosado, A.</dc:creator>
<dc:creator>Sanchez, L.</dc:creator>
<dc:creator>Torroja, C.</dc:creator>
<dc:creator>Dopazo, A.</dc:creator>
<dc:creator>Sanchez-Cabo, F.</dc:creator>
<dc:creator>Hidalgo, A.</dc:creator>
<dc:creator>Senders, M. L.</dc:creator>
<dc:creator>van Leent, M. M. T.</dc:creator>
<dc:creator>Seep, L.</dc:creator>
<dc:creator>Hasenauer, J.</dc:creator>
<dc:creator>Placek, K.</dc:creator>
<dc:creator>Thiele, C.</dc:creator>
<dc:creator>Netea, M. G.</dc:creator>
<dc:creator>Mulder, W. J. M.</dc:creator>
<dc:creator>Riksen, N. P.</dc:creator>
<dc:creator>Perez-Medina, C.</dc:creator>
<dc:date>2025-11-30</dc:date>
<dc:identifier>doi:10.1101/2025.11.26.689708</dc:identifier>
<dc:title><![CDATA[HDL-associated phosphatidylserine blunts myeloid activation and protects from atherosclerosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.26.690792v1?rss=1">
<title>
<![CDATA[
Uncovering senescent fibroblast heterogeneity connects DNA damage response to idiopathic pulmonary fibrosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.26.690792v1?rss=1</link>
<description><![CDATA[
Cellular senescence is a largely heterogeneous state of cell stress that deleteriously accumulates with age. Many types of heterogeneity in senescence have been described; however, cellular senescence within the same cell type has only started to be documented. Here, we show primary, human lung fibroblasts from donors who are healthy or diagnosed with idiopathic pulmonary fibrosis (IPF) exhibit a subtle form of heterogeneity over time after DNA damage. Moreover, senescent IPF lung fibroblasts display a dysregulated transcriptional-protein DNA damage response (DDR). Weighted gene correlation network analysis (WGCNA) reveals unique and known targets linking senescent IPF lung fibroblast heterogeneity to genes associated with DNA damage and repair, cytokine and chemokine responses, and extracellular matrix (ECM) signaling. We combine our healthy and IPF senescent gene expression signatures to develop a novel gene set of senescence-associated genes that identify disease-relevant cells in human single-cell RNA-seq (scRNA-seq) data. Collectively, our results uncover human-relevant senescence signatures, highlight IPF-specific DDR, cytokine and chemokine, and ECM targets, and expand our understanding of how a dysregulated DDR contributes to senescent cell heterogeneity in IPF.
]]></description>
<dc:creator>Hughes, J.-W. B.</dc:creator>
<dc:creator>Pujari, A.</dc:creator>
<dc:creator>Sandholm, A.</dc:creator>
<dc:creator>Croll, D.</dc:creator>
<dc:creator>Monk, L. B.</dc:creator>
<dc:creator>Joshua, I.</dc:creator>
<dc:creator>Butterfield, R.</dc:creator>
<dc:creator>Horton, C.</dc:creator>
<dc:creator>Schneider, K.</dc:creator>
<dc:creator>Senchyna, F.</dc:creator>
<dc:creator>Brown, I.</dc:creator>
<dc:creator>Coelho, A. L.</dc:creator>
<dc:creator>Ho, T.-C.</dc:creator>
<dc:creator>Deguchi, H.</dc:creator>
<dc:creator>Le Saux, C. J.</dc:creator>
<dc:creator>Deinhardt-Emmer, S.</dc:creator>
<dc:creator>Ellerby, L. M.</dc:creator>
<dc:creator>Vitari, A.</dc:creator>
<dc:creator>Furman, D.</dc:creator>
<dc:creator>Hogoboam, C. M.</dc:creator>
<dc:creator>Laslavic, A.</dc:creator>
<dc:creator>Desprez, P.-Y.</dc:creator>
<dc:creator>Quarta, M.</dc:creator>
<dc:creator>Campisi, J.</dc:creator>
<dc:date>2025-11-30</dc:date>
<dc:identifier>doi:10.1101/2025.11.26.690792</dc:identifier>
<dc:title><![CDATA[Uncovering senescent fibroblast heterogeneity connects DNA damage response to idiopathic pulmonary fibrosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.26.690767v1?rss=1">
<title>
<![CDATA[
Beyond Correlation: An Ultra-Large Physics-Driven Vascularized Tumor Model to Bridge Feature Formation with Underlying Biology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.26.690767v1?rss=1</link>
<description><![CDATA[
Radiomics provides an appealing, non-invasive approach to probing tumor biology for potential diagnostic and prognostic applications. However, its clinical adoption is limited by challenges in interpretability, which in turn compromise its robustness. To uncover the underlying causation, we developed an ultra-large-scale (ULS) computational model that simulates heterogeneous, vascularized tumor growth under physical constraints to a scale that can be visualized in medical images. Our study revealed the pivotal role of tumor proliferation rate in driving necrosis and tissue heterogeneity and the dominant impact of oxygen consumption rate on vascularization level. Analysis of the resultant tumor Radiomics shows a causal relationship between tumor biophysical parameters and imaging features. Specifically, differences in proliferation and oxygen consumption rates result in distinct changes in radiomic image features, identifying suitable imaging modalities and quantitative imaging metrics for studying these biophysical parameters. This work thus reverse-engineers the building blocks of Radiomics as a means to understand their respective biological underpinnings.
]]></description>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Jin, L.</dc:creator>
<dc:creator>Sheng, K.</dc:creator>
<dc:date>2025-12-01</dc:date>
<dc:identifier>doi:10.1101/2025.11.26.690767</dc:identifier>
<dc:title><![CDATA[Beyond Correlation: An Ultra-Large Physics-Driven Vascularized Tumor Model to Bridge Feature Formation with Underlying Biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.26.690816v1?rss=1">
<title>
<![CDATA[
GPR34 regulation of disease-associated microglial states and responses to physiological stimuli 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.26.690816v1?rss=1</link>
<description><![CDATA[
GPR34 is a microglia-enriched GPCR whose expression is downregulated under several disease conditions, including Alzheimers disease (AD) and multiple sclerosis (MS). Despite this, its function is poorly understood in normal or disease conditions, as is its contribution to disease-related microglia states. Using RNA-sequencing, we find that microglia from global Gpr34 knockout (KO) mouse brains exhibited transcriptional shifts toward disease-associated microglia (DAM) and inflammatory profiles, partially mirroring the microglial phenotype seen in 5xFAD AD model mice. Notably, when Gpr34 KO mice were crossed with 5xFAD mice, DAM transcriptional profiles and glial pathology were further exacerbated despite the already robust DAM signature driven by amyloidosis. This occurred without affecting amyloid plaque burden. In human stem cell-derived microglia (iMGLs), GPR34 KO strongly reduced calcium (Ca{superscript 2}) and phosphorylated ERK (pERK) signaling in response to known GPR34 agonists, including lyso-phosphatidylserine (lysoPS) and myelin, and caused transcriptional alterations linked to immune regulation and cell proliferation. Interestingly, GPR34 loss selectively impaired phagocytosis of myelin but not amyloid-{beta} or E. coli. Furthermore, GRP34 KO diminished, but did not abolish, the transcriptional response elicited by myelin. Together, these findings suggest that GPR34 is important for maintaining microglia homeostasis, mediating phagocytosis of and transcriptional response to myelin, and restraining microglial response to neurodegenerative disease conditions.
]]></description>
<dc:creator>Geller, A.</dc:creator>
<dc:creator>Kwon, M. J.</dc:creator>
<dc:creator>Simmons, S. K.</dc:creator>
<dc:creator>Xu, Q.</dc:creator>
<dc:creator>Martenis, W. E.</dc:creator>
<dc:creator>Doman, J.</dc:creator>
<dc:creator>Natarajan, S.</dc:creator>
<dc:creator>Stalnaker, K. J.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Adiconis, X.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Demers, M.</dc:creator>
<dc:creator>Batzli, D.</dc:creator>
<dc:creator>Valle-Tojeiro, A.</dc:creator>
<dc:creator>Biji, C.</dc:creator>
<dc:creator>Aryal, S.</dc:creator>
<dc:creator>Pribiag, H.</dc:creator>
<dc:creator>Depp, C.</dc:creator>
<dc:creator>Morshed, N.</dc:creator>
<dc:creator>Misri, D.</dc:creator>
<dc:creator>Bohannon, D.</dc:creator>
<dc:creator>Longhi, E.</dc:creator>
<dc:creator>Keshishian, H.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>McKinney, D.</dc:creator>
<dc:creator>Gould, A. E.</dc:creator>
<dc:creator>Lebois, E.</dc:creator>
<dc:creator>Weiwer, M.</dc:creator>
<dc:creator>Johnson, M.</dc:creator>
<dc:creator>Levin, J. Z.</dc:creator>
<dc:creator>Zhang, Y.-L.</dc:creator>
<dc:creator>Sheng, M.</dc:creator>
<dc:creator>Kunwar, P. S.</dc:creator>
<dc:date>2025-12-01</dc:date>
<dc:identifier>doi:10.1101/2025.11.26.690816</dc:identifier>
<dc:title><![CDATA[GPR34 regulation of disease-associated microglial states and responses to physiological stimuli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.28.691229v1?rss=1">
<title>
<![CDATA[
FCHo2, instead of talin, enables inside-out activation of integrin αvβ5 in curved adhesions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.28.691229v1?rss=1</link>
<description><![CDATA[
Inside-out activation of integrins is crucial for transducing mechanical forces through the extracellular ligand-integrin-talin-F-actin axis. Extensive studies have shown that talin is the essential player in this process by binding to the intracellular tail of {beta} integrins. Here, we show that, while talin binding is essential for inside-out integrin activation in focal adhesions, it is dispensable in curved adhesions - a distinct adhesion architecture that is exclusively mediated by integrin v{beta}5 and selectively formed at curved membranes. Instead, a curvature-sensing protein FCHo2 binds to the HDRRE motif in the cytoplasmic tail of integrin {beta}5 (ITG{beta}5) and inside-out activates integrin v{beta}5 in curved adhesions. Intriguingly, FCHo2 does not bind to a similar motif in the homologous integrin {beta}3 tail. Through truncations and mutations, we identified a pivotal tryptophan (W) in the {beta}3 tail, which is conserved in all homologous integrin {beta} isoforms except {beta}5, where it is replaced by a tyrosine (Y766). This tyrosine substitution is crucial for integrin {beta}5s unique capability in forming curved adhesions. A Y766W mutation abolishes integrin {beta}5s capacity to form curved adhesions, without affecting its ability to form focal adhesions. Furthermore, our studies suggest that the phosphorylation state of Y766 regulates whether integrin v{beta}5 forms curved adhesions or focal adhesions, providing a cellular mechanism governing different adhesion types. Overall, our work unveils distinct molecular interactions and regulatory mechanisms between curved adhesions and focal adhesions, and establishes a molecular basis for the formation of curved adhesions by integrin v{beta}5.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=108 SRC="FIGDIR/small/691229v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Lu, C.-H.</dc:creator>
<dc:creator>Lee, C. E.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Valencia, L. A.</dc:creator>
<dc:creator>You, H.</dc:creator>
<dc:creator>Tsai, C.-T.</dc:creator>
<dc:creator>Cui, B.</dc:creator>
<dc:date>2025-12-01</dc:date>
<dc:identifier>doi:10.1101/2025.11.28.691229</dc:identifier>
<dc:title><![CDATA[FCHo2, instead of talin, enables inside-out activation of integrin αvβ5 in curved adhesions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.30.691411v1?rss=1">
<title>
<![CDATA[
Interrogating the Regulatory Function of HAQERs during Human Cortical Development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.30.691411v1?rss=1</link>
<description><![CDATA[
BackgroundSequence divergence within gene regulatory elements has been proposed to play an important role in the evolution of human-specific traits, including cortical expansion. However, the mutational processes that efficiently modify gene regulatory elements and the target genes upon which they act are poorly understood. We investigated the regulatory function and origins of the fastest evolved regions in the human genome, termed Human Ancestor Quickly Evolved Regions (HAQERs), in their native genomic context during human cerebral cortex development.

ResultsWe identified 50 HAQERs with accessible chromatin in developing human cortex, largely arising from previously unconstrained ancestral sequences. To test the necessity of these HAQERs for gene regulation, we established an all-in-one CRISPRi lentiviral vector and linked 26 HAQERs to nearby target genes across cell types and Wnt pathway activation contexts. Rapid gains of CpGs distinguished HAQERs active during cortical development and displaying human-specific epigenomic marks. As a high density of CpG sites can drive formation of permissive chromatin, we identified 107 HAQERs with at least 17 human-specific CpG gains per kb, termed HAQER CpG Beacons. These HAQERs emerged via contributions from GC-biased gene conversion (gBGC) with evidence for selection preferentially fixing CpG sites. Notably, the CHL1 and DPP10 loci, both implicated in human neurological disorders, each harbored two gene-linked HAQER CpG Beacons.

ConclusionsOur findings reveal HAQER target genes and support a model where gBGC and natural selection jointly drive regulatory-altering CpG variants to fixation, forging regulatory innovations in the human cortex.
]]></description>
<dc:creator>Abeykoon, Y.</dc:creator>
<dc:creator>Dzikowski, N.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Sinniah, E.</dc:creator>
<dc:creator>Weaver, S.</dc:creator>
<dc:creator>Pavlovic, B. J.</dc:creator>
<dc:creator>Wallace, J. L.</dc:creator>
<dc:creator>Mangan, R. J.</dc:creator>
<dc:creator>Lowe, C. B.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:date>2025-12-01</dc:date>
<dc:identifier>doi:10.1101/2025.11.30.691411</dc:identifier>
<dc:title><![CDATA[Interrogating the Regulatory Function of HAQERs during Human Cortical Development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.30.691432v1?rss=1">
<title>
<![CDATA[
Bacterial strains in the human gut associate with host physiology 
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</title>
<link>https://biorxiv.org/content/10.1101/2025.11.30.691432v1?rss=1</link>
<description><![CDATA[
The human microbiota influences host physiology, yet much of its complexity lies beneath the species level. Here, we analyzed the intra-species genetic diversity of 936 gut bacteria across 24,997 individuals from three countries. Our findings show that highly abundant species exhibit greater strain stability, whereas low-abundance species display increased in-clonal mutations. Clonal strains are often mutually exclusive, while genetically variable strains tend to coexist. Strain turnover is associated with the presence of annotated chemotaxis and sporulation genes in reference genomes, whereas strain coexistence is associated with quorum sensing and secretion systems. Leveraging deep phenotypic data, we constructed an atlas detailing strain-level associations with diverse host physiological domains. For example, Phocaeicola vulgatus sub-types relate to host obesity, Lachnospira eligens to sleep, and Parabacteroides distasonis to iron hemostasis. This resource may guide personalized microbiome-based interventions to improve human health.
]]></description>
<dc:creator>Shoer, S.</dc:creator>
<dc:creator>Godneva, A.</dc:creator>
<dc:creator>Weinberger, A.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Pilpel, Y.</dc:creator>
<dc:creator>Segal, E.</dc:creator>
<dc:date>2025-12-01</dc:date>
<dc:identifier>doi:10.1101/2025.11.30.691432</dc:identifier>
<dc:title><![CDATA[Bacterial strains in the human gut associate with host physiology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.11.29.691326v1?rss=1">
<title>
<![CDATA[
Need-selective gating of dopamine neuron cue responses by real and virtual hunger 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.11.29.691326v1?rss=1</link>
<description><![CDATA[
The midbrain dopamine system is important for linking reward-predictive cues to learning and 20 motivation. Here we investigated how dopamine neuron responses to food and water cues are modulated by changes in internal state. We developed a flexible cued-approach task that allowed us to examine behavioral and neural responses to both food- and water-predictive cues within the same recording session. We found that overlapping subsets of dopamine neurons respond to food and water cues, but that the magnitude of these responses is gated in a need-25 specific way. Stimulation of hunger-promoting AgRP neurons amplified dopamine neuronresponses to food cues, but not water cues, and the magnitude of these responses exceeded those observed in natural hunger. These findings indicate that changes in internal state modulate, in a need-appropriate way, the responses of a common set of dopamine neurons to environmental signals of food and water availability.
]]></description>
<dc:creator>Mamaligas, A. A.</dc:creator>
<dc:creator>Berke, J.</dc:creator>
<dc:creator>Kreitzer, A. C.</dc:creator>
<dc:creator>Knight, Z. A.</dc:creator>
<dc:date>2025-12-02</dc:date>
<dc:identifier>doi:10.64898/2025.11.29.691326</dc:identifier>
<dc:title><![CDATA[Need-selective gating of dopamine neuron cue responses by real and virtual hunger]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.11.30.691330v1?rss=1">
<title>
<![CDATA[
An orthogonal TRAP enables intersectional genetic access to activated neurons in the mouse brain 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.11.30.691330v1?rss=1</link>
<description><![CDATA[
The study of neural circuits has been greatly enabled by methods for obtaining genetic access to activated neurons. However, these approaches typically tag neurons based on their response to only a single stimulus, which limits the ability to define precise subpopulations of cells. Here we describe an approach (X-TRAP) in which the activity-dependent expression of Flp recombinase is gated by branaplam, a small molecule that triggers splicing of the X-ON switch. We show that X-TRAP knock-in mice exhibit undetectable Flp recombination in the absence of drug and that branaplam treatment results in robust induction of recombination selectively in neurons that express FOS. Moreover, we show that X-TRAP is orthogonal to the widely-used TRAP system, such that these two approaches can be used in the same animal to label cells with Cre and Flp recombination in response to two different stimuli. We apply this strategy to map neural circuits that control food intake. This approach for intersectional, activity-dependent genetic labeling should enhance our ability to identify the neural correlates of behavior.
]]></description>
<dc:creator>Chatziris, N.</dc:creator>
<dc:creator>Jarvie, B. C.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Knight, Z. A.</dc:creator>
<dc:date>2025-12-02</dc:date>
<dc:identifier>doi:10.64898/2025.11.30.691330</dc:identifier>
<dc:title><![CDATA[An orthogonal TRAP enables intersectional genetic access to activated neurons in the mouse brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.11.30.691333v1?rss=1">
<title>
<![CDATA[
Cellular coding of ingestion in the caudal brainstem 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.11.30.691333v1?rss=1</link>
<description><![CDATA[
The termination of a meal is triggered by sensory feedback from the stomach and intestines that reports on ingested food1-10 and is relayed to the caudal nucleus of the solitary tract (cNTS) in the brainstem11-14. This sensory feedback is thought to gradually intensify as a meal progresses, resulting in the progressive activation of cNTS circuits that promote satiety15,16, but this idea has never been tested by recording the single-cell activity of cNTS neurons while animals eat. Here, we have used a preparation for calcium imaging in the caudal brainstem of behaving animals17 to characterize how food ingestion is encoded in the cNTS. We find that when food is delivered directly to the stomach, most cNTS neurons exhibit ramping activation over many minutes that tracks cumulative food consumed and depends on canonical gut-brain pathways. However, when the same food is consumed by mouth, this widespread ramping activation is gone, and most cNTS neurons instead exhibit phasic, seconds-timescale responses to oral contact with food. We show that these rapid responses are driven by a combination of mechanical, gustatory and nutritive signals from the mouth and throat and do not require traditional gut-brain pathways, including gut-innervating vagal afferents, although GI feedback modulates their duration. We show that one source of this rapid input is descending projections from the paraventricular hypothalamus, which track ingestion dynamics and are required for proper meal termination. These findings reveal that sensory feedback from the stomach and intestines, which is directly transmitted to the cNTS and thought to be critical for satiety, is not the major driver of cNTS activity during ingestion. Instead, these circuits make extensive use of rapid, pregastric signals that report on the dynamics of behavior.
]]></description>
<dc:creator>Ly, T.</dc:creator>
<dc:creator>Yi, X.</dc:creator>
<dc:creator>Lee, G. R.</dc:creator>
<dc:creator>Grove, J. C.</dc:creator>
<dc:creator>Sibih, Y. E.</dc:creator>
<dc:creator>Oh, Y. Y.</dc:creator>
<dc:creator>Qiu, L.</dc:creator>
<dc:creator>Sivakumar, N.</dc:creator>
<dc:creator>Knight, Z. A.</dc:creator>
<dc:date>2025-12-02</dc:date>
<dc:identifier>doi:10.64898/2025.11.30.691333</dc:identifier>
<dc:title><![CDATA[Cellular coding of ingestion in the caudal brainstem]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.11.30.691441v1?rss=1">
<title>
<![CDATA[
A genetic screen in enteroendocrine cells reveals mechanisms that control protein sensing and GLP-1 release 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.11.30.691441v1?rss=1</link>
<description><![CDATA[
Enteroendocrine cells (EECs) are the principal nutrient sensors in the gastrointestinal (GI) tract and release hormones such as glucagon-like-peptide 1 (GLP-1) that modulate GI function and appetite. While some of the molecules involved in nutrient sensing within EECs have been described, there have been no systematic studies to map the relevant genes and pathways. Here, we developed a strategy to perform a high-throughput screen for genes that are required for nutrient-induced activation of EECs, and we applied this to probe mechanisms for sensing dietary protein. We found that all of the genes previously proposed to function as protein sensors in EECs are, collectively, dispensable for protein sensing in an EEC cell line. Instead, a screen of >20,000 sgRNAs identified numerous genes associated with mitochondrial respiration as being necessary for this process. We showed through secondary assays that impairing oxidative phosphorylation (OXPHOS) reduced EEC activation and GLP-1 release in response to nutrients but not in response to a non-nutritive stimulus. On the other hand, boosting OXPHOS increased EEC activation and GLP-1 release. These data reveal that intracellular metabolism within EECs controls the detection of dietary protein, possibly by monitoring the entry of ingested amino acids into the TCA cycle. More broadly, these findings suggest a general strategy to screen for genes and pathways that might be used to boost the nutrient-regulated release of gut peptides such as GLP-1.
]]></description>
<dc:creator>Yu, S.</dc:creator>
<dc:creator>Lee, Y.</dc:creator>
<dc:creator>Boggess, S. C.</dc:creator>
<dc:creator>Klein, N. R.</dc:creator>
<dc:creator>Teyssier, N.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Knight, Z. A.</dc:creator>
<dc:date>2025-12-02</dc:date>
<dc:identifier>doi:10.64898/2025.11.30.691441</dc:identifier>
<dc:title><![CDATA[A genetic screen in enteroendocrine cells reveals mechanisms that control protein sensing and GLP-1 release]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.01.687254v1?rss=1">
<title>
<![CDATA[
Advancing High-Resolution 7T Diffusion MRI: Evaluating Phase-Encoding Correction Strategies for Distortion Correction from Basic to Four-Way Acquisitions 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.01.687254v1?rss=1</link>
<description><![CDATA[
PurposeHigh-resolution 7T diffusion MRI (dMRI) is limited by image artifacts that compromise anatomical accuracy. The purpose of this study was to systematically evaluate phase-encoding (PE) acquisition and correction strategies to determine which methods best mitigate geometric distortions and improve data reproducibility.

MethodsFive healthy adults were each scanned twice on a 7T MRI scanner (0.9 mm isotropic resolution), using a highly oversampled dMRI protocol with four PE directions (AP, PA, RL, LR). From this dataset, we created and processed eleven time-equivalent, 10-minute acquisitions, ranging from uncorrected single-PE data to comprehensive 4-way PE schemes. These strategies were quantitatively compared on their geometric alignment with T1-weighted images and on the scan-rescan reproducibility of DTI-derived metrics.

Results(1) All distortion-corrected schemes significantly improved geometric accuracy over uncorrected data; (2) Strategies correcting with a full set of reversed-PE (2-way) diffusion weighted images (DWIs) outperformed the common approach of using only a single reversed b=0 image; and (3) a 4-way PE acquisition consistently provided the highest image fidelity and reproducibility. The optimized acquisition enabled high-quality reconstruction of both long-range and fine-scale superficial white matter pathways.

ConclusionFor high-resolution 7T dMRI, multi-PE acquisition is essential to achieve accurate geometry and stable microstructural estimates (i.e., less residual EPI distortion and better scan-rescan agreement). A 4-way PE scheme provides the most accurate and reproducible results for microstructural and connectivity modeling.

Data statementData will be made available in BIDS format upon acceptance of the manuscript. To be updated with DOI.
]]></description>
<dc:creator>Schilling, K. G.</dc:creator>
<dc:creator>Beckett, A. J. S.</dc:creator>
<dc:creator>Amandola, M.</dc:creator>
<dc:creator>Walker, E. B.</dc:creator>
<dc:creator>Feinberg, D. A.</dc:creator>
<dc:creator>Bunge, S. A.</dc:creator>
<dc:creator>Vu, A. T.</dc:creator>
<dc:date>2025-12-03</dc:date>
<dc:identifier>doi:10.64898/2025.12.01.687254</dc:identifier>
<dc:title><![CDATA[Advancing High-Resolution 7T Diffusion MRI: Evaluating Phase-Encoding Correction Strategies for Distortion Correction from Basic to Four-Way Acquisitions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.03.689730v1?rss=1">
<title>
<![CDATA[
Female-predominant anti-CD4 IgG autoantibody production and its correlations with plasma levels of progesterone, microbial translocation, and blunted immune reconstitution in HIV-infected patients on suppressive ART 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.03.689730v1?rss=1</link>
<description><![CDATA[
Autoimmunity contributes to HIV immunopathogenesis even in the absence of overt autoimmune disease. We previously showed that anti-CD4 autoantibodies from people with HIV (PWH) on suppressive antiretroviral therapy (ART) can mediate cytotoxicity against CD4+ T cells, implicating a role in impaired immune reconstitution. Despite viral suppression, many PWH with poor CD4 recovery exhibit chronic immune activation, microbial translocation, and dysregulated humoral immunity. Here, we identify a female-predominant elevation of plasma anti-CD4 IgG autoantibodies in aviremic PWH receiving ART. Across two independent cohorts, HIV-positive females, but not males, displayed significantly higher anti-CD4 IgG, predominantly IgG1, compared with HIV-negative controls, without parallel increases in anti-CD4 IgA or IgM. This sex-specific pattern was unique to anti-CD4 IgG and was not observed for anti-CD8 IgG, anti-double-stranded DNA IgG, or anti-nuclear antigen IgG; these control autoantibodies correlated with one another but not with anti-CD4 IgG. Elevated anti-CD4 IgG levels were associated with lower plasma progesterone levels and reduced absolute CD4+ T-cell counts. Markers of microbial translocation, soluble CD14 (sCD14), lipopolysaccharide-binding protein (LBP), and lipopolysaccharide (LPS), were also selectively increased in HIV-positive females, with sCD14 and LBP showing significant or borderline associations with anti-CD4 IgG. Together, these findings identify anti-CD4 IgG as a sex-dimorphic autoimmune signature in treated HIV infection, linked to progesterone levels, persistent microbial translocation, and incomplete immune recovery. This work highlights an under-recognized intersection of sex, mucosal barrier dysfunction, and autoimmunity in HIV pathogenesis and suggests potential therapeutic targets to improve immune reconstitution in women.
]]></description>
<dc:creator>McKinnon, J. E.</dc:creator>
<dc:creator>Wan, Z.</dc:creator>
<dc:creator>Luo, Z.</dc:creator>
<dc:creator>Hartley, A.</dc:creator>
<dc:creator>Price, R. W.</dc:creator>
<dc:creator>Gisslen, M.</dc:creator>
<dc:creator>Jiang, W.</dc:creator>
<dc:date>2025-12-04</dc:date>
<dc:identifier>doi:10.64898/2025.12.03.689730</dc:identifier>
<dc:title><![CDATA[Female-predominant anti-CD4 IgG autoantibody production and its correlations with plasma levels of progesterone, microbial translocation, and blunted immune reconstitution in HIV-infected patients on suppressive ART]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.04.691947v1?rss=1">
<title>
<![CDATA[
The Emergence of Novel Versus Known Three-Dimensional Structures from Random Sequences. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.04.691947v1?rss=1</link>
<description><![CDATA[
It has been hypothesized that while random sequences are unlikely to fold into proteins of the length of globular proteins, repeated random sequences are more likely to adopt stably folded structures, with implications for molecular evolution. We used structure prediction methods to determine the foldability of approximately 120-residue sequences composed of 5-to 60-residue random repeats. With repeats of less than 30-residues, sequences were frequently discovered (1-12%) that fold with high confidence. For less than 60-residue repeats, we frequently observe {beta}-solenoids, similar to those seen in natural proteins. We observe solenoids stabilized by apolar packing as well as ones stabilized by polar interactions with Ca2+ in the core of the structure as in natural RTX domains. Helical bundles were observed with high frequency when insertions or deletions (INDELs) were included between blocks of repeating sequences. We also observed a new super-secondary structure consisting of a tightly wound -helical screw, and experimentally confirmed its stability and structure by CD spectroscopy and X-ray crystallography. Thus, structure predictors can discover structures that are well out of the distribution of the data upon which they were trained. Beyond 40-residue repeat lengths very few sequences were predicted to fold. The small number of structures we observed were representative of well-established major classes of tertiary structures; greater sampling would be needed to discover novel structures from a random distribution. These studies illuminate dark matter regions of protein structure space and support previous predictions that proteins evolved through the assortment of shorter peptide sequences.

Significance statementThe availability of powerful and accurate programs for predicting protein three-dimensional structures enables one to ask fundamental questions concerning the origin of folded functional proteins during evolution. We show that 120-residue proteins composed of random sequences repeated in tandem are predicted to be much more likely to fold than fully random proteins. These studies validate previous predictions that proteins evolved through the repetition and assortment of short peptide sequences. Also, some of the predicted structures represent novel conformations, which were confirmed experimentally. These findings advance our understanding of molecular evolution and have implications for design of novel proteins.
]]></description>
<dc:creator>Yang, R. S.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Davydenko, A.</dc:creator>
<dc:creator>Mawaldi, Z.</dc:creator>
<dc:creator>Kormos, R. C.</dc:creator>
<dc:creator>Myerscough, D.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>DeGrado, W.</dc:creator>
<dc:date>2025-12-04</dc:date>
<dc:identifier>doi:10.64898/2025.12.04.691947</dc:identifier>
<dc:title><![CDATA[The Emergence of Novel Versus Known Three-Dimensional Structures from Random Sequences.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.03.691944v1?rss=1">
<title>
<![CDATA[
Tri-AD: Hippocampal cell-type-specific responses to age, sex and APOE genotype 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.03.691944v1?rss=1</link>
<description><![CDATA[
Alzheimers disease (AD) risk is strongly shaped by age, sex, and the apolipoprotein E {varepsilon}4 (APOE4) allele--the strongest genetic risk factor for late-onset AD. While each factor has been studied independently, their combined impact on cellular and molecular processes remains unclear. Here, we used single-nucleus RNA sequencing (snRNA-seq) to profile hippocampal cell states in a sex-balanced cohort of human APOE4/4 and APOE3/3 knock-in mice across 6, 12, and 18 months of age. We identify sex as the major driver of variation in cell-type abundance and find that oligodendrocytes exhibit pronounced male-biased transcriptional sensitivity to APOE4. Differential expression and cell-cell communication analyses further reveal sex-divergent temporal trajectories in inhibitory neurons, with females showing early APOE4-associated suppression of synaptic pathways and males displaying a delayed but convergent decline. Together, these findings clarify how age, sex, and APOE genotype jointly regulate cell-type-specific gene expression and intercellular communication in the aging hippocampus, providing an innovative and publicly accessible database for aging and AD research and related precision medicine.
]]></description>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Pereda Serras, C.</dc:creator>
<dc:creator>Yao, L.</dc:creator>
<dc:creator>Liang, Z.</dc:creator>
<dc:creator>Almonte-Loya, A.</dc:creator>
<dc:creator>Blumenfeld, J.</dc:creator>
<dc:creator>Suan, K.</dc:creator>
<dc:creator>Yoon, S. Y.</dc:creator>
<dc:creator>Grone, B.</dc:creator>
<dc:creator>Ding, L.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Sun, S.</dc:creator>
<dc:creator>Oskotsky, B.</dc:creator>
<dc:creator>Oskotsky, T.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2025-12-08</dc:date>
<dc:identifier>doi:10.64898/2025.12.03.691944</dc:identifier>
<dc:title><![CDATA[Tri-AD: Hippocampal cell-type-specific responses to age, sex and APOE genotype]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.03.692228v1?rss=1">
<title>
<![CDATA[
Senescence-Linked Fibrosis in the Aging Human Ovary Revealed by p16-Based Histological Profiling and Spatial Transcriptomics 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.03.692228v1?rss=1</link>
<description><![CDATA[
Cellular senescence is implicated as a driver of ovarian aging, but senescent cells in the human postmenopausal ovary remain poorly defined. Using spatially resolved p16INK4a protein expression, a canonical senescence marker, we identified and mapped senescent cells in postmenopausal ovaries. We integrated p16 immunohistochemistry, multiplexed immunofluorescence, spatial transcriptomics, and AI-guided digital pathology to map senescent microenvironments. p16-positive cells formed discrete stromal, vascular, and cyst-associated clusters that increased with age and were enriched for macrophages and myofibroblast-like cells. Wholetranscriptome profiling of 92 spatial regions uncovered a 32-gene p16-associated signature, BuckSenOvary, that distinguished p16-positive regions across cortex and medulla. BuckSenOvary is characterized by suppression of cell-cycle regulators and activation of inflammatory and extracellular-matrix remodelling genes. AI-based collagen matrix analysis confirmed that p16-positive regions exhibit more architecturally complex collagen, demonstrating that focal senescent microenvironments are fibro-inflammatory. These findings position senescent ovarian niches as therapeutic targets to preserve ovarian function.
]]></description>
<dc:creator>Watson, M. A.</dc:creator>
<dc:creator>Devrukhkar, P. R.</dc:creator>
<dc:creator>Murad, N. F.</dc:creator>
<dc:creator>Wu, F.</dc:creator>
<dc:creator>Kim, M. J.</dc:creator>
<dc:creator>Anvari, H.</dc:creator>
<dc:creator>Tran, U.</dc:creator>
<dc:creator>Martin, N.</dc:creator>
<dc:creator>Tran, T.</dc:creator>
<dc:creator>Zaza, G.</dc:creator>
<dc:creator>Schneider, K.</dc:creator>
<dc:creator>Soygur, B.</dc:creator>
<dc:creator>Shanes, E. D.</dc:creator>
<dc:creator>Wirtz, D.</dc:creator>
<dc:creator>Pavone, M. G.</dc:creator>
<dc:creator>Melov, S.</dc:creator>
<dc:creator>Wu, P.-H.</dc:creator>
<dc:creator>Furman, D.</dc:creator>
<dc:creator>Duncan, F. E.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:date>2025-12-08</dc:date>
<dc:identifier>doi:10.64898/2025.12.03.692228</dc:identifier>
<dc:title><![CDATA[Senescence-Linked Fibrosis in the Aging Human Ovary Revealed by p16-Based Histological Profiling and Spatial Transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.06.689939v1?rss=1">
<title>
<![CDATA[
Human stem cell models for group 3 medulloblastoma uncover JARID1B as a regulator of the chromatin landscape. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.06.689939v1?rss=1</link>
<description><![CDATA[
Medulloblastoma (MB) is one of the most prevalent malignant brain tumors in children, with tremendous cognitive and neuroendocrine disability among survivors. Group 3 MB (G3MB) has poor overall survival at <50%, high frequencies of metastases, and no targeted therapies. Amplification of MYC and activation of TGF{beta} signaling occur frequently in G3MB. Many tumors have no reported mutations, suggesting epigenetic drivers. We here describe novel humanized models for G3MB from human induced pluripotent stem cells (hiPSC). By transducing hiPSC-derived neuroepithelial stem cells (NESC), we determined that: 1) both MYC and TGF{beta} effectors drove tumors in vivo; 2) MYC/TGF{beta}R1 in combination led to more aggressive tumors and resistance to clinical inhibitors of TGF{beta}, and 3) NESC-derived tumors clustered with human G3MB. To decipher mechanisms, we integrated RNA-sequencing with CUT&RUN (for MYC genomic localization and post-translational modification of histones). MYC-bound neural developmental genes were repressed in MYC/TGF{beta}R1 co-driven lines. Gene signatures associated with the Polycomb Repressive Complex (PRC) demarcated with H3K27me3; the histone mark directly regulated by PRC. We identified JARID1B, a MYC binding partner and H3K4me3 demethylase, as a regulator of repressed neural genes. Primary G3MB also showed increased levels of H3K27me3 concurrent with higher expression of JARID1B. Knockdown of JARID1B in human G3MB cell lines reduced growth, supporting potential as a therapeutic target. We conclude that a MYC-TGF{beta}-JARID1B axis represses target genes to drive G3MB and present new humanized models for G3MB to understand epigenetic dysregulation in G3MB.
]]></description>
<dc:creator>Qadeer, Z. A.</dc:creator>
<dc:creator>Hou, E.</dc:creator>
<dc:creator>Siegel, D.</dc:creator>
<dc:creator>Westelman, S.</dc:creator>
<dc:creator>Gudenas, B.</dc:creator>
<dc:creator>Smith, K.</dc:creator>
<dc:creator>Ismael, W. M.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Johnson, M. O.</dc:creator>
<dc:creator>Grele, S.</dc:creator>
<dc:creator>Phua, R.</dc:creator>
<dc:creator>Hendrikse, L.</dc:creator>
<dc:creator>Guney, E.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Gaspar-Maia, A.</dc:creator>
<dc:creator>Northcott, P. A.</dc:creator>
<dc:creator>Taylor, M. D.</dc:creator>
<dc:creator>Weiss, W. A.</dc:creator>
<dc:date>2025-12-08</dc:date>
<dc:identifier>doi:10.64898/2025.12.06.689939</dc:identifier>
<dc:title><![CDATA[Human stem cell models for group 3 medulloblastoma uncover JARID1B as a regulator of the chromatin landscape.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.04.692390v1?rss=1">
<title>
<![CDATA[
Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.04.692390v1?rss=1</link>
<description><![CDATA[
Apolipoprotein E4 (APOE4) confers the greatest genetic risk for developing Alzheimers disease (AD). With APOE4 broadly expressed in the brain, its cell-type-specific roles in AD pathogenesis are only beginning to be defined. Here, we show that neuronal APOE4 expression drives damaging lipid accumulation in hippocampal neurons, oligodendrocytes, and microglia, with preferential buildup of peroxidized lipids in microglia in a tauopathy mouse model. Neuron-specific removal of APOE4 abolished this lipid phenotype, whereas neuron-specific expression of APOE4 was sufficient to recapitulate it, demonstrating that neuronal APOE4 is both necessary and sufficient for lipid accumulation. Strikingly, the association between lipid burden, microgliosis, and neurodegeneration was strongest in mice with neuron-specific APOE4 expression. Single-nucleus RNA sequencing revealed neuronal APOE4-vulnerable neuron populations, as well as enrichment of disease-associated microglia and oligodendrocytes, all promoting lipid pathology. Primary mouse co-culture experiments showed that neuronal APOE4 drives microglial lipid accumulation via contact-dependent mechanisms involving uptake of lipids from neurons and oligodendrocytes. These findings establish neuronal APOE4 as a key driver of lipid accumulation via neuron-oligodendrocyte-microglia interactions, providing mechanistic insight into APOE4-driven lipid pathology in AD.
]]></description>
<dc:creator>Kim, M. J.</dc:creator>
<dc:creator>Blumenfeld, J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Yip, O.</dc:creator>
<dc:creator>Yao, L.</dc:creator>
<dc:creator>Ancheta, S.</dc:creator>
<dc:creator>De Leon, S.</dc:creator>
<dc:creator>Platow, Z.</dc:creator>
<dc:creator>Liang, Z.</dc:creator>
<dc:creator>Shostak, D.</dc:creator>
<dc:creator>Suan, K.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Koutsodendris, N.</dc:creator>
<dc:creator>Ellis, C.</dc:creator>
<dc:creator>Nguyen, J.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:date>2025-12-09</dc:date>
<dc:identifier>doi:10.64898/2025.12.04.692390</dc:identifier>
<dc:title><![CDATA[Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.05.692665v1?rss=1">
<title>
<![CDATA[
Allele Frequencies at Recessive Disease Genes are Mainly Determined by Pleiotropic Effects in Heterozygotes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.05.692665v1?rss=1</link>
<description><![CDATA[
The classic theory of mutation-selection balance predicts the equilibrium frequency of genetic variation under negative selection. The model predicts a simple relationship between the total frequency of deleterious variants, mutation rate, and strength of selection, with different functions for recessive and (co-)dominant genes. In this study, we investigate whether genes associated with human recessive disorders fit the predictions of this classic model. By comparing observed frequencies of loss of function variants (LoFs) to those expected under mutation-selection balance we find that, for nearly all recessive genes, the observed frequencies are too low to be explained by purely recessive selection. Analyzing the effects of heterozygous LoFs on quantitative traits from the UK Biobank, we find that recessive disease genes have widespread quantitative effects in heterozygotes. Together, these results suggest that most selection experienced by pathogenic mutations in recessive disease genes may be due to stabilizing selection in heterozygotes. We conclude that very few human genes follow the classic model of recessive mutation-selection balance.
]]></description>
<dc:creator>Judd, J.</dc:creator>
<dc:creator>Spence, J. P.</dc:creator>
<dc:creator>Milind, N.</dc:creator>
<dc:creator>Kachuri, L.</dc:creator>
<dc:creator>Witte, J. S.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:date>2025-12-09</dc:date>
<dc:identifier>doi:10.64898/2025.12.05.692665</dc:identifier>
<dc:title><![CDATA[Allele Frequencies at Recessive Disease Genes are Mainly Determined by Pleiotropic Effects in Heterozygotes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.07.692769v1?rss=1">
<title>
<![CDATA[
Inhibitory potential of autologous neutralizing antibodies sets quantitative limits on the rebound-competent HIV-1 reservoir 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.07.692769v1?rss=1</link>
<description><![CDATA[
HIV-1 cure requires preventing viral rebound after treatment interruption, but quantitative criteria defining the rebound-competent reservoir are lacking. We studied individuals undergoing observational treatment interruption to identify virologic and immunologic determinants of rebound. In 9 of 13 participants, rebound viruses were genetically identical or similar to proviruses in circulating resting CD4 T-cells. We found no evidence of recombination among rebound sequences, rather resistance to autologous neutralizing antibodies was a critical determinant of viral rebound. Using inhibitory potential (IP), the log reduction in single-round infection at physiologic IgG concentrations, we defined quantitative limits governing rebound-competency. Reservoir variants exhibited a wide range of IP values (0.4-8.2 logs), whereas rebound viruses were minimally inhibited (0.5-2.8 logs), indicating that inhibition by even up to 2.8 logs (631-fold) cannot prevent rebound. Longitudinal analyses revealed that waning aNAb potency allows previously neutralized variants to gain rebound potential. Thus, rebound competency is a dynamic, immune-governed property defined by quantitative immunologic constraints.
]]></description>
<dc:creator>Garcia, M. A.</dc:creator>
<dc:creator>Farrell-Sherman, A.</dc:creator>
<dc:creator>Zhuo, J.</dc:creator>
<dc:creator>Fray, E. J.</dc:creator>
<dc:creator>Zinsser, A. M.</dc:creator>
<dc:creator>Aydin, B.</dc:creator>
<dc:creator>Sowers, K.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Lopez, B. M.</dc:creator>
<dc:creator>Abeyta-Lopez, A.</dc:creator>
<dc:creator>Chu, T.</dc:creator>
<dc:creator>Lubbeck, D.</dc:creator>
<dc:creator>Chae, M.</dc:creator>
<dc:creator>Varriale, J.</dc:creator>
<dc:creator>Westfall, D. H.</dc:creator>
<dc:creator>Lai, J.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Dalhuisen, T.</dc:creator>
<dc:creator>Simonetti, F. R.</dc:creator>
<dc:creator>Peluso, M. J.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Siliciano, R. F.</dc:creator>
<dc:creator>Cohn, L. B.</dc:creator>
<dc:creator>Siliciano, J. D.</dc:creator>
<dc:date>2025-12-09</dc:date>
<dc:identifier>doi:10.64898/2025.12.07.692769</dc:identifier>
<dc:title><![CDATA[Inhibitory potential of autologous neutralizing antibodies sets quantitative limits on the rebound-competent HIV-1 reservoir]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.08.693033v1?rss=1">
<title>
<![CDATA[
Alveolar Type II Cell-derived MMP1high basal cells promote destructive microcysts in idiopathic pulmonary fibrosis. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.08.693033v1?rss=1</link>
<description><![CDATA[
Idiopathic Pulmonary Fibrosis (IPF) is a fatal lung disease characterized by progressive epithelial metaplasia and widespread fibrosis. Alveolar microcysts develop near terminal airways in IPF and are linked to poor outcome. Using HTII-280 as a short-term lineage marker of AT2-derived AT0 (SFTPC+/SCGB3A2+) and basaloid (KRT17) cells, together with organoids and spatial transcriptomics (Xenium), we highlight epithelial similarities between respiratory bronchioles (RBs) and alveolar microcysts both having AT0, SCGB3A2+, and basaloid/basal cells (BCs), albeit with expanded BCs in IPF microcysts. The AT0- and AT2-derived BCs strongly express the collagenase, matrix metalloproteinase protein-1, MMP1 in organoids -- mirroring in situ BCs lining IPF microcysts, but distinct from MMP1low BCs in large airways or normal lungs. Expression of MMP1 correlates with basal cell hypoxia pathway activity.

MMP1high AT2-derived BCs and IPF BCs promoted type 1 collagen degradation ex vivo and in vivo after xenotransplantation, forming microcystic structures that were abrogated by concurrent MMP inhibitor treatment. Notably, a Frizzled 5 WNT agonist antibody reversed the MMP1high state of AT2-derived BCs, raising a possible therapeutic approach. These findings suggest AT2 transdifferentiation to basaloid/basal cells is uncommon in normal lungs but can expand as a potential source of alveolar destruction, likely contributing to the pernicious course of IPF.
]]></description>
<dc:creator>Garakani, K.</dc:creator>
<dc:creator>Brumwell, A. N.</dc:creator>
<dc:creator>Toigo, M.</dc:creator>
<dc:creator>Jacob, A.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Ho, T. C.</dc:creator>
<dc:creator>Wolters, P.</dc:creator>
<dc:creator>Matthay, M. A.</dc:creator>
<dc:creator>Volk, R. F.</dc:creator>
<dc:creator>Zaro, B. W.</dc:creator>
<dc:creator>Iantorno, S. A.</dc:creator>
<dc:creator>Berhan, A.</dc:creator>
<dc:creator>Sheppard, D. W.</dc:creator>
<dc:creator>Auyeung, V. C.</dc:creator>
<dc:creator>Jourdan Le Saux, C. C.</dc:creator>
<dc:creator>Chapman, H. A.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.08.693033</dc:identifier>
<dc:title><![CDATA[Alveolar Type II Cell-derived MMP1high basal cells promote destructive microcysts in idiopathic pulmonary fibrosis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.09.693260v1?rss=1">
<title>
<![CDATA[
Comparing Multislice Simulations of MD Simulations with CryoEM Exposes Membrane Prediction Errors 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.09.693260v1?rss=1</link>
<description><![CDATA[
Cryo-electron microscopy (cryoEM) is a powerful tool for atomic- and molecular-resolution structure determination, while molecular dynamics (MD) simulations are similarly powerful tools for predicting molecular trajectories. Given the challenges in estimating biomolecule dynamics with cryoEM alone, MD simulations are employed to forecast molecular motions and to interpret cryoEM reconstructions. Few methods, however, can evaluate MD predictions directly. Here, we use multislice wave propagation to project sampled snapshots of MD trajectories, either coarse-grained (CG) or all-atom (AA), into simulated cryoEM 3D reconstructions. We compared simulated and experimental images of low- and high-curvature membranes to show that MD simulations qualitatively reflect the fluidity and thus the contrast of biological membranes observed by cryoEM. MD simulations also correctly predicted bilayer dimensions for single component flat bilayers observed in cryoEM images. However, Martini3 CG-MD simulations failed to predict changes in membrane thickness induced by high curvature and with heterogeneous lipid compositions. We pinpointed the misbehavior of polyunsaturated lipid tails and cholesterol in Martini3 simulations as the main error sources contributing to inaccurate bilayer thicknesses. Our comparisons also explain membrane structure discrepancies between cryoEM and small angle X-ray scattering (SAXS). Further testing of MD predictions by direct comparisons between simulated and experimental cryoEM images should lead to the development of more accurate MD force fields.

Statement of SignificanceMolecular dynamics (MD) simulations are frequently employed to predict the dynamics of biological macromolecules and assemblies, but these predictions remain difficult to validate experimentally. Cryo-electron microscopy (cryoEM) can be used to directly image the conformational ensemble of macromolecules, but images of Coulombic potential cannot be easily compared to snapshots of atoms from MD simulations. Here, we show that a physics-based multislice image projection algorithm accurately converts MD trajectories to simulated cryoEM 2D images and 3D reconstructions. Using this approach, we identify consistencies and discrepancies between MD simulations and cryoEM experiments. Notably, coarse-grained MD performs poorly compared to all-atom MD when simulating membranes composed of mixtures of lipids that include cholesterol and polyunsaturated lipids, providing observables for MD force field improvement.
]]></description>
<dc:creator>Mohammed, A.</dc:creator>
<dc:creator>Lincoff, J.</dc:creator>
<dc:creator>Natale, A.</dc:creator>
<dc:creator>Ophus, C.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>Frost, A.</dc:creator>
<dc:creator>Moss, F. R.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.09.693260</dc:identifier>
<dc:title><![CDATA[Comparing Multislice Simulations of MD Simulations with CryoEM Exposes Membrane Prediction Errors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.10.693544v1?rss=1">
<title>
<![CDATA[
Activation of cannabinoid receptor CB1 leads to aberrant myelination in development 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.10.693544v1?rss=1</link>
<description><![CDATA[
The endocannabinoid system (ECS) has a widespread role in the development and function of the central nervous system (CNS). Cannabinoid receptors like CB1 and CB2 can be activated with exogenous cannabinoids most popularly known as tetrahydrocannabinol (THC) or cannabis and cannabidiol (CBD). The components of the ECS are expressed early in fetal development, and prenatal exposure to cannabis can lead to structural changes in white matter. White matter is composed of neuronal axons ensheathed in myelin, a lipid-rich insulation that facilitates saltatory conduction and maintains axon integrity. In the CNS, myelin is made by specialized glial cells called oligodendrocytes (OLs), which in addition to neurons also express components of the ECS. However, while several studies have focused on how the ECS regulates neuronal development, there is a limited understanding of its impact on OL development or myelin formation.

Therefore, our current study set out to understand how pharmacological activation of the ECS alters OL differentiation and myelin formation in vivo. We administered WIN 55,212-2 (WIN 55), a CB1 and CB2 agonist, to larval zebrafish and longitudinally analyzed OL development and myelination in vivo. Interestingly, we observed an increase in non-axonal ensheathments in the spinal cord, which appeared to be surrounding neuronal cell bodies. These non-axonal ensheathments were dependent on CB1, as the addition of WIN 55 in a global CB1 mutant prevented this phenotype. Furthermore, this ectopic cell body ensheathment occurred independently from normal myelination processes, as individual OLs did not exhibit changes in the number of myelin sheaths, sheath length, or total myelin output. This study shows that activation of CB receptors in vivo leads to increased non-axonal ensheathment without significantly changing OL differentiation or normal myelin formation. Future studies can further investigate the pathways that drive this phenotype to better understand how exogenous cannabinoid activation can regulate the precision of oligodendrocyte ensheathment.
]]></description>
<dc:creator>Miramontes, T. G.</dc:creator>
<dc:creator>Hamling, K. R.</dc:creator>
<dc:creator>Doan, R. A.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Collins, H. Y.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:creator>Call, C. L.</dc:creator>
<dc:creator>Monk, K.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.10.693544</dc:identifier>
<dc:title><![CDATA[Activation of cannabinoid receptor CB1 leads to aberrant myelination in development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.09.693258v1?rss=1">
<title>
<![CDATA[
Poorer physical function is associated with elevated spatial entropy in the aging brain network landscape 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.09.693258v1?rss=1</link>
<description><![CDATA[
Life is a constant struggle against disorder. As we age, our ability to maintain internal order declines. In the healthy human brain, order is observable in the form of functionally segregated brain network communities that exhibit spatial consistency. These communities associate with distinct cognitive and physical functions. When mapped into the brain, they form a functional "landscape". We assessed the spatial disorder of these landscapes in older adults with a wide range of mobility using a modified version of Shannon entropy. We found that compared to younger adults, older adults had significantly higher entropy in the sensorimotor cortex, basal ganglia, hippocampus, thalamus, and occipital lobe. Higher entropy in many of these regions was associated with worse physical function and higher body mass index in older adults. Findings suggest that spatial entropy in brain network landscapes may be a marker of declining physical function. Modifiable factors, such as losing excess weight, may help to ameliorate aging-related brain entropy increases in regions such as the sensorimotor cortex, which may in turn help to preserve physical function in older adults.
]]></description>
<dc:creator>McIntyre, C. C.</dc:creator>
<dc:creator>ODonnell, S. M.</dc:creator>
<dc:creator>Lyday, R. G.</dc:creator>
<dc:creator>Burdette, J. H.</dc:creator>
<dc:creator>Cummings, S. R.</dc:creator>
<dc:creator>Kritchevsky, S. B.</dc:creator>
<dc:creator>Laurienti, P. J.</dc:creator>
<dc:date>2025-12-12</dc:date>
<dc:identifier>doi:10.64898/2025.12.09.693258</dc:identifier>
<dc:title><![CDATA[Poorer physical function is associated with elevated spatial entropy in the aging brain network landscape]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.09.693329v1?rss=1">
<title>
<![CDATA[
Mapping the circulating proteome across neurodegeneration: A harmonized, consortium-scale framework for uncovering molecular pathophysiology 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.09.693329v1?rss=1</link>
<description><![CDATA[
Large-scale plasma proteomics offers unprecedented opportunities to investigate the systemic biology of neurodegeneration, yet technical heterogeneity, site-specific artifacts, and clinical confounding remain major barriers to reproducible discovery. Leveraging data from 13,733 individuals with Alzheimers disease (AD), Parkinsons disease (PD), frontotemporal dementia (FTD), Parkinsons disease dementia (PDD), amyotrophic lateral sclerosis (ALS), and non-impaired controls in the Global Neurodegeneration Proteomics Consortium (GNPC), we present a scalable and generalizable analytical framework for harmonizing and interpreting consortium-scale proteomic datasets. Using a high-dimensional perturbation framework, we systematically benchmark five commonly used batch correction methods across a range of realistic confounding structures, including site-disease imbalance, nonlinear effects, and heteroskedasticity. Empirical Bayes modelling via limma consistently emerged as the most robust method, optimally balancing removal of site-related technical variance with retention of disease-relevant biological signal. On this harmonized foundation, we resolve neurodegenerative disease plasma signatures, including a shared immune-metabolic axis in AD and PD, neuromuscular disruption in ALS, and proteostatic imbalance in PD. Tissue and cell-type enrichment highlight widespread immune-endocrine involvement in AD and hematopoietic activation in PD. Demographically matched analyses nominate distinct, candidate biomarkers across diseases, including lipid, redox, and complement factors in AD, lysosomal and cytoskeletal proteins in PD, and muscle-derived markers in ALS. This study establishes a scalable analytical framework for integrating real-world proteomic data and provides a disease-resolved catalogue of circulating signatures to inform biomarker development and targeted intervention across neurodegenerative diseases.
]]></description>
<dc:creator>Finney, C. A.</dc:creator>
<dc:creator>An, L.</dc:creator>
<dc:creator>Winchester, L. M.</dc:creator>
<dc:creator>Vogel, J.</dc:creator>
<dc:creator>Wilkins, H. M.</dc:creator>
<dc:creator>Burns, J. M.</dc:creator>
<dc:creator>Swerdlow, R. H.</dc:creator>
<dc:creator>Slawson, C.</dc:creator>
<dc:creator>Rothstein, J. D.</dc:creator>
<dc:creator>Global Neurodegeneration Proteomics Consortium,</dc:creator>
<dc:creator>Lutz, M. W.</dc:creator>
<dc:creator>Saloner, R.</dc:creator>
<dc:creator>Shvetcov, A.</dc:creator>
<dc:date>2025-12-12</dc:date>
<dc:identifier>doi:10.64898/2025.12.09.693329</dc:identifier>
<dc:title><![CDATA[Mapping the circulating proteome across neurodegeneration: A harmonized, consortium-scale framework for uncovering molecular pathophysiology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.10.693551v1?rss=1">
<title>
<![CDATA[
Bifunctional transcriptional effector domains control gene expression pulses in an occupancy-dependent manner 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.10.693551v1?rss=1</link>
<description><![CDATA[
Dynamic gene expression pulses enable adaptive response to stimuli and can be generated in natural and synthetic systems. Controlling these dynamics typically involves circuits consisting of multiple genes and transcription factors (TFs). Here, we discover a new class of bifunctional transcriptional effector domains that can first activate and subsequently repress the same gene, producing dynamic gene expression pulses from a single input. These pulse dynamics arise from distinct, temporally separated chromatin states defined by active and repressive chromatin modifications. The balance between active and repressed states is determined by the DNA occupancy of the bifunctional TF. Bifunctional domains activate at low occupancy but switch to repression at high occupancy, resulting in a non-monotonic TF input-gene expression output relationship tunable by TF concentration and number of DNA binding sites. We develop a kinetic model that links TF occupancy to gene expression transitions, allowing for the programming of eight distinct cell "states" - combinations of On/Off states of 3 reporter genes - using a single bifunctional effector. This work establishes the theoretical framework and molecular mechanisms of pulse-generating gene regulation by bifunctional domains and creates a foundation for engineering complex multi-gene circuits.
]]></description>
<dc:creator>Andrews, C. J.</dc:creator>
<dc:creator>Costa, E. J.</dc:creator>
<dc:creator>Janer Carattini, G. L.</dc:creator>
<dc:creator>DelRosso, N. V.</dc:creator>
<dc:creator>Fujimori, T.</dc:creator>
<dc:creator>Shimasawa, M.</dc:creator>
<dc:creator>Bintu, L.</dc:creator>
<dc:date>2025-12-12</dc:date>
<dc:identifier>doi:10.64898/2025.12.10.693551</dc:identifier>
<dc:title><![CDATA[Bifunctional transcriptional effector domains control gene expression pulses in an occupancy-dependent manner]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.10.693468v1?rss=1">
<title>
<![CDATA[
Neuronal APOE4 reduction with an APOE-I3-targeting ASO protects against neurodegeneration and neuroinflammation in an Alzheimer's disease mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.10.693468v1?rss=1</link>
<description><![CDATA[
Apolipoprotein E4 (APOE4) is the strongest genetic risk factor for late-onset Alzheimers disease (AD). Within the central nervous system (CNS), APOE is produced by a variety of cell types, with differential roles in AD pathogenesis. Studies have shown that APOE4 produced by neurons plays a central role in promoting the development of major AD pathologies, including p-tau accumulation, neuroinflammation, and neurodegeneration, highlighting its role as an upstream initiating factor that affects other cell types and downstream AD-related pathologies. Here, we demonstrate that antisense oligonucleotides (ASOs) targeting APOE-I3, a neuron-specific splicing variant of APOE mRNA, effectively reduce APOE expression in neurons in vitro and in vivo. Treating PS19 tauopathy mice expressing APOE4 with this APOE-I3-targeting ASO reduces neuronal APOE4, rescues neurodegeneration, and diminishes neuroinflammation. Strikingly, the extent of neuronal APOE4 reduction predicts the efficacy of rescuing neurodegeneration. Single nucleus RNA-sequencing demonstrated that APOE-I3-targeting ASO treatment decreases disease associated neuronal and glial subtypes and increases a disease-protective microglial subtype. These findings suggest that preferential knockdown of neuronal APOE4 with an APOE-I3-targeting ASO protects against key hallmarks of AD pathology, elucidating a potential therapeutic approach for treating APOE4-driven AD.
]]></description>
<dc:creator>Yip, O.</dc:creator>
<dc:creator>Yao, L.</dc:creator>
<dc:creator>Blumenfeld, J.</dc:creator>
<dc:creator>Kim, M. J.</dc:creator>
<dc:creator>Shostak, D.</dc:creator>
<dc:creator>Platow, Z.</dc:creator>
<dc:creator>Suan, K.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Ellis, C.</dc:creator>
<dc:creator>An, A.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Xu, Q.</dc:creator>
<dc:creator>De Leon, S.</dc:creator>
<dc:creator>Nguyen, J.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:date>2025-12-13</dc:date>
<dc:identifier>doi:10.64898/2025.12.10.693468</dc:identifier>
<dc:title><![CDATA[Neuronal APOE4 reduction with an APOE-I3-targeting ASO protects against neurodegeneration and neuroinflammation in an Alzheimer's disease mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.10.693598v1?rss=1">
<title>
<![CDATA[
MechanoMR microparticle (M3) sensors reveal dynamic stress loading as a driver of epithelial-mesenchymal transition 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.10.693598v1?rss=1</link>
<description><![CDATA[
The dynamic mechanical response of tissues underlies their physiological function, yet direct, quantitative measurement of tissue stress in vivo has remained a major challenge. Here, we introduce the mechanoMR microparticle (M3, "M-cube") sensor, a hybrid soft-matter/nanoparticle probe that integrates directly into tissue mechanical networks while transducing local stress into quantitative magnetic resonance (MR) readouts with single-particle resolution. We demonstrate the utility of this platform across diverse model systems, including tumor spheroids, Xenopus embryos, and mouse xenografts, where the M3 sensor enables noninvasive, spatiotemporally resolved mapping of tissue stress dynamics during cancer development. Using this approach, we reveal that epithelial-mesenchymal transition (EMT) is accompanied by distinctive stress-remodeling patterns observable in vivo. Strikingly, we find that abrupt stress increases, rather than cumulative or peak stress magnitude, are the key determinants of EMT induction in cancer cells within the tumor microenvironment. Transcriptomic profiling under controlled stress-loading dynamics shows that sustained yet gradual stress escalation activates cytoprotective antioxidation pathways (e.g., FOXO/AMPK) that reinforce epithelial stability, whereas acute stress surges overwhelm these defense mechanisms, predisposing cells toward mesenchymal reprogramming. These findings establish the M3 sensor as a broadly applicable technology for linking dynamic mechanical cues to cell-state transitions in development, homeostasis, and disease.
]]></description>
<dc:creator>An, M.</dc:creator>
<dc:creator>Yu, R.</dc:creator>
<dc:creator>Lin, A.</dc:creator>
<dc:creator>Jang, J.</dc:creator>
<dc:creator>Ohk, J.</dc:creator>
<dc:creator>Kim, Y.-J.</dc:creator>
<dc:creator>Jung, H.</dc:creator>
<dc:creator>Kwak, M.</dc:creator>
<dc:creator>Jun, Y.-w.</dc:creator>
<dc:creator>Cheon, J.</dc:creator>
<dc:date>2025-12-13</dc:date>
<dc:identifier>doi:10.64898/2025.12.10.693598</dc:identifier>
<dc:title><![CDATA[MechanoMR microparticle (M3) sensors reveal dynamic stress loading as a driver of epithelial-mesenchymal transition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.13.694145v1?rss=1">
<title>
<![CDATA[
Degraded sensory coding in a mouse model of Scn2a-related disorder and its rescue by CRISPRa gene activation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.13.694145v1?rss=1</link>
<description><![CDATA[
Heterozygous loss-of-function mutations in SCN2A, a sodium channel gene expressed in cortical pyramidal (PYR) cells, lead to a neurodevelopmental disorder characterized by autism, intellectual disability, and cortical sensory dysfunction. In Scn2a+/- mice, PYR dendritic excitability and synapses are impaired, but cortical information processing deficits are unknown. In the whisker somatosensory cortex, we found strongly degraded somatotopic tuning of PYR cells, profoundly blurred whisker maps, and impaired population coding, despite normal overall firing rates. This constitutes a robust biomarker for Scn2a-related cortical dysfunction. Parvalbumin (PV) interneurons were also unexpectedly hypofunctional. We tested for rescue of coding deficits in post-critical period, young adult mice by viral delivery of CRISPR activation (CRISPRa) reagents that upregulate Scn2a. CRISPRa treatment normalized cortical sensory representations at the single-unit and map levels. This suggests that therapy to increase Scn2a expression may be effective in normalizing cortical function in Scn2a loss-of-function disorder, even in older children or adults.
]]></description>
<dc:creator>Vandemark, K.</dc:creator>
<dc:creator>Monday, H. R.</dc:creator>
<dc:creator>Rodriguez, L.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Lee, M.</dc:creator>
<dc:creator>Ahituv, A.</dc:creator>
<dc:creator>Satapathy, A.</dc:creator>
<dc:creator>Page, N. F.</dc:creator>
<dc:creator>An, K.</dc:creator>
<dc:creator>Hamada, E. C.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Bender, K. J.</dc:creator>
<dc:creator>Feldman, D. E.</dc:creator>
<dc:date>2025-12-14</dc:date>
<dc:identifier>doi:10.64898/2025.12.13.694145</dc:identifier>
<dc:title><![CDATA[Degraded sensory coding in a mouse model of Scn2a-related disorder and its rescue by CRISPRa gene activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.14.692434v1?rss=1">
<title>
<![CDATA[
Virus-like particles enable targeted gene engineering and pooled CRISPR screening in primary human myeloid cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.14.692434v1?rss=1</link>
<description><![CDATA[
Primary human myeloid cells are promising candidates for immunotherapy, yet efficient and scalable technologies for genetic engineering and screening in these cells are limited. Here we present a virus-like particle (VLP)-based toolkit that delivers diverse CRISPR genome editing modalities to human monocytes, macrophages, and dendritic cells with high efficiency while preserving viability and innate immune responsiveness. VLP-mediated delivery of ribonucleoprotein payloads supports gene knockout, base editing and epigenetic silencing, and enables site-specific integration of large DNA sequences when combined with AAV donors for homology-directed repair. Leveraging sgRNA delivery via VPX-lentivirus combined with Cas9 protein delivery via engineered virus-like particle (eVLP) treatment ("SLICeVLP"), we performed the first pooled loss-of-function screens in human macrophages. We uncovered regulators of TNF production and CD80 expression in human macrophages, converging on TNFAIP3 as a central regulator of inflammatory polarization. TNFAIP3 ablation promoted a pro-inflammatory cell state that is resistant to suppressive polarization, and augmented cytotoxicity of engineered HER2 CAR-macrophages. Taken together, this technology platform enables unbiased discovery and characterization of functional gene targets in primary human myeloid cells.
]]></description>
<dc:creator>Jung, H.</dc:creator>
<dc:creator>Devant, P.</dc:creator>
<dc:creator>Ching, C.</dc:creator>
<dc:creator>Ota, M.</dc:creator>
<dc:creator>Hamilton, J.</dc:creator>
<dc:creator>Steinhart, Z.</dc:creator>
<dc:creator>Ngo, W.</dc:creator>
<dc:creator>Sandoval, L.</dc:creator>
<dc:creator>Jung, J. H.</dc:creator>
<dc:creator>Xu, D.</dc:creator>
<dc:creator>An, M.</dc:creator>
<dc:creator>Urs, E.</dc:creator>
<dc:creator>Chen, P. A.</dc:creator>
<dc:creator>Allain, V.</dc:creator>
<dc:creator>Tada, T.</dc:creator>
<dc:creator>Nunez, J. K.</dc:creator>
<dc:creator>Landau, N. R.</dc:creator>
<dc:creator>Liu, D. R.</dc:creator>
<dc:creator>Eyquem, J.</dc:creator>
<dc:creator>Doudna, J. A.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Carnevale, J.</dc:creator>
<dc:date>2025-12-14</dc:date>
<dc:identifier>doi:10.64898/2025.12.14.692434</dc:identifier>
<dc:title><![CDATA[Virus-like particles enable targeted gene engineering and pooled CRISPR screening in primary human myeloid cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.12.693854v1?rss=1">
<title>
<![CDATA[
Rotational settings quantize nucleosome movement by chromatin regulators 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.12.693854v1?rss=1</link>
<description><![CDATA[
Proper nucleosome positioning is essential for gene regulation and genomic integrity. Regulated nucleosome assembly and positioning results from a need to protect DNA sequences genome-wide, constrained by the known intrinsic sequence preferences of histones. Current models posit that chromatin regulators override the intrinsic preferences to establish the nucleosome landscapes observed in vivo, implying minimal roles for DNA sequence in guiding nucleosomal structure in cells. In contrast, we demonstrate that DNA sequence intrinsically guides the structure and remodeling of nucleosomes from yeast to mammals. We demonstrate that nucleosomes with weak translational settings in vitro, in yeast, and in mammalian cells demonstrate a clear preference for inward-facing A/T dinucleotides and outward facing G/C dinucleotides, at 10 bp spacings consistent with established preferences for rotationally positioned nucleosomes. Foreign DNA sequences heterologously inserted into the yeast genome obey similar rules, indicating that DNA sequence itself is causal. Finally, remodelers and transcription elongation change the preference among the alternative translational positions 10 bp apart, retaining the rotational setting. From these results, we propose that DNA sequence creates an energy landscape with preferred rotational settings every [~]10 bp, and that chromatin regulators, rather than overriding these preferences, navigate within them. This  quantized ratchet model provides a unifying framework for understanding how diverse cellular processes achieve precise nucleosome positioning while maintaining the same DNA face exposed to regulatory factors.
]]></description>
<dc:creator>La, V.</dc:creator>
<dc:creator>Trouth, A.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Ramachandran, S.</dc:creator>
<dc:date>2025-12-15</dc:date>
<dc:identifier>doi:10.64898/2025.12.12.693854</dc:identifier>
<dc:title><![CDATA[Rotational settings quantize nucleosome movement by chromatin regulators]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.12.694063v1?rss=1">
<title>
<![CDATA[
Integrative Genomic and Functional Analyses Reveal NINL as a Modulator of Tau Aggregation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.12.694063v1?rss=1</link>
<description><![CDATA[
IntroductionProteostasis dysfunction is a hallmark of frontotemporal dementia (FTD) and Alzheimers disease (AD), yet the genetic and molecular pathways that disrupt protein homeostasis remain poorly understood.

MethodsWe integrated human genetics, transcriptomics, and functional studies to identify proteostasis network components involved in tauopathy.

ResultsWe identified 18 proteostasis network genes harboring 75 rare, damaging variants enriched in FTD and/or AD. These genes, spanning multiple proteostasis pathways, were differentially expressed in MAPT mutant neurons and dysregulated in FTD and AD brains. NINL, which encodes Nlp, emerged as the only gene consistently upregulated across all datasets. NINL overexpression reduced tau seeding and enhanced lysosomal proteolytic activity, whereas two FTD-enriched NINL frameshift variants impaired Nlp expression and abolished these protective effects.

DiscussionOur findings identify a set of proteostasis genes with genetic and transcriptional links to neurodegeneration and reveal NINL as a novel regulator of tau aggregation, potentially upregulated as an adaptive response to proteotoxic stress.
]]></description>
<dc:creator>Swift, S. K.</dc:creator>
<dc:creator>Huang, G.</dc:creator>
<dc:creator>Cochran, J. N.</dc:creator>
<dc:creator>Minaya, M. A.</dc:creator>
<dc:creator>Castruita, P. A.</dc:creator>
<dc:creator>Miller, K. J.</dc:creator>
<dc:creator>Starr, E.</dc:creator>
<dc:creator>Galasso, G.</dc:creator>
<dc:creator>Marsh, J. A.</dc:creator>
<dc:creator>Kao, A.</dc:creator>
<dc:creator>Yokoyama, J. S.</dc:creator>
<dc:creator>Karch, C. M.</dc:creator>
<dc:date>2025-12-16</dc:date>
<dc:identifier>doi:10.64898/2025.12.12.694063</dc:identifier>
<dc:title><![CDATA[Integrative Genomic and Functional Analyses Reveal NINL as a Modulator of Tau Aggregation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.14.694179v1?rss=1">
<title>
<![CDATA[
The Binding Landscape of an Essential Nuclear Hormone Receptor in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.14.694179v1?rss=1</link>
<description><![CDATA[
Transcription factors (TFs) control gene expression by binding to response elements but distinguishing functional binding sites from non-functional ones remains a major challenge in the field of transcriptional regulation. To probe transcription factor binding and function in a multicellular organism we characterized the binding and transcriptional landscape of NHR-25, a highly-conserved and essential transcription factor in C. elegans. Using CRISPR/Cas9, we tagged the essential nuclear hormone receptor, nhr-25, with GFP and FLAG at its endogenous locus in C. elegans and performed ChIP-seq. We found that NHR-25 binds with two distinct modes: proximal promoter binding and distal enhancer-type binding. Motif analysis of the ChIP-seq peaks showed that the NHR-25 binding motif depended on chromatin context and identified additional enriched motifs, suggesting other putative regulatory partners at composite response elements. By combining ChIP-seq with conditional RNAi knockdown, we characterized nhr-25s transcriptome and identified putative direct targets of NHR-25 binding. Through an 11-bp mutation of the summit of an identified NHR-25 occupied region, we demonstrated that a single response element was responsible for regulating the expression of multiple genes. Our findings provide a comprehensive view of an essential transcription factors functional binding landscape at native protein levels, highlighting the intricate nature of response element logic and demonstrating the critical role of a single regulatory element in regulating gene expression.
]]></description>
<dc:creator>Sinks, A. L.</dc:creator>
<dc:creator>Asahina, M.</dc:creator>
<dc:creator>Yamamoto, K. R.</dc:creator>
<dc:creator>Ward, J. D.</dc:creator>
<dc:creator>Thurtle-Schmidt, D. M.</dc:creator>
<dc:date>2025-12-16</dc:date>
<dc:identifier>doi:10.64898/2025.12.14.694179</dc:identifier>
<dc:title><![CDATA[The Binding Landscape of an Essential Nuclear Hormone Receptor in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.15.694494v1?rss=1">
<title>
<![CDATA[
Functional diversification of the cephalopod proteome by RNA-editing 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.15.694494v1?rss=1</link>
<description><![CDATA[
Coleoid cephalopods exhibit the highest levels of ADAR-mediated RNA editing of any known animal, yet the functional consequences of most recoding events remain largely unknown. We integrate proteomics with biochemical and cellular assays to characterize thousands of recoding events across the Doryteuthis pealeii proteome. Using quantitative and functional mass spectrometry, we show that RNA edit-driven recoding reshapes the cellular proteome to alter protein stability, subcellular localization, post-translational modifications, and enzymatic activity. --Recoding can regulate post-translational modifications through their creation or ablation, and this has direct effects on protein function and protein-protein interactions. Recoding of the E3 ligase MARCHF5 drives widespread changes in substrate ubiquitylation and perturbs mitochondrial homeostasis, illustrating how RNA editing can influence organelle function. These data provide the first proteome-scale view of how extensive RNA recoding diversifies protein function in coleoid cephalopods and offers a new framework for understanding how RNA-level plasticity shapes protein function and cellular physiology.
]]></description>
<dc:creator>Moen, J. M.</dc:creator>
<dc:creator>Richards, A. L.</dc:creator>
<dc:creator>Krogsaeter, E. K.</dc:creator>
<dc:creator>Polacco, B.</dc:creator>
<dc:creator>Gordon, M.</dc:creator>
<dc:creator>Stevenson, E.</dc:creator>
<dc:creator>Boyles, R.</dc:creator>
<dc:creator>Burq, M.</dc:creator>
<dc:creator>Stepec, D.</dc:creator>
<dc:creator>Crampton, B.</dc:creator>
<dc:creator>Cimermancic, P.</dc:creator>
<dc:creator>Rosenthal, J. J. C.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Swaney, D. L.</dc:creator>
<dc:date>2025-12-16</dc:date>
<dc:identifier>doi:10.64898/2025.12.15.694494</dc:identifier>
<dc:title><![CDATA[Functional diversification of the cephalopod proteome by RNA-editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.16.694760v1?rss=1">
<title>
<![CDATA[
Mapping of critical prosodic and phonetic networks in post-stroke apraxia of speech 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.16.694760v1?rss=1</link>
<description><![CDATA[
PurposeMany have made proposals to better diagnose and/or classify post-stroke apraxia of speech (AOS), with some arguing for the separation of AOS into behavioral subtypes. Recent studies of primary progressive AOS have promoted a separation of prosodic and phonetic subtypes, aligning with a dual-motor coordination model separating the neural substrates of prosodic and phonetic function. Motivated by the limited corroboration of these subtypes in post-stroke AOS, here we present mapping results in a cohort of stroke survivors aiming to identify distinct neural substrates for prosodic and phonetic aspects of speech motor coordination.

MethodsLeft-hemisphere stroke survivors (n = 127; 64 with AOS) received speech-language evaluation and neuroimaging at the Center for the Study and Treatment of Aphasia Recovery (C-STAR). AOS severity was quantified via the Apraxia of Speech Rating Scale (ASRS). We utilized a novel lesion-symptom mapping technique with an emphasis on prediction that identifies ensembles of regions supporting performance in the prosodic and phonetic domains.

ResultsAn ensemble of networks supporting prosodic function localized to dorsal and ventral (but primarily dorsal) sensorimotor cortex, as well as a distributed network of white matter pathways connecting Rolandic cortex to auditory regions and cerebellum, emphasizing the role of auditory feedback processing and laryngeal control in supporting prosodic function. A separate but partially overlapping network supporting phonetic function localized primarily to ventral Rolandic cortex and the arcuate fasciculus.

ConclusionsThis work represents the first mapping of prosodic and phonetic subtypes in post-stroke AOS in a large cohort of individuals. We hope our results motivate the development of assessment and treatment techniques individually targeting prosodic and phonetic functioning to better serve individuals with AOS and facilitate clinical discussion of the disorder.
]]></description>
<dc:creator>Kurteff, G. L.</dc:creator>
<dc:creator>Walker, G. M.</dc:creator>
<dc:creator>Fridriksson, J.</dc:creator>
<dc:creator>Hickok, G.</dc:creator>
<dc:date>2025-12-19</dc:date>
<dc:identifier>doi:10.64898/2025.12.16.694760</dc:identifier>
<dc:title><![CDATA[Mapping of critical prosodic and phonetic networks in post-stroke apraxia of speech]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.16.694766v1?rss=1">
<title>
<![CDATA[
Mu opioid receptor mRNA and protein localization across the rat and mouse habenula 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.16.694766v1?rss=1</link>
<description><![CDATA[
The habenula (Hb), which contains medial and lateral subdivisions (MHb and LHb, respectively), has high intensity mu opioid binding and receptor (MOR) expression, yet the details of MOR localization across these regions remains debated. MHb and LHb participate in largely non-overlapping neural circuits, therefore accurately resolving MOR expression across MHb and LHb is critical for understanding how MOR ligands impact behaviors. Here we utilized in situ hybridization (ISH) and immunocytochemistry (ICC) to systematically map Oprm1 mRNA and MOR protein throughout the habenular complex. We studied both rat and mouse tissue to evaluate conserved expression across two common research species. Importantly, we found mRNA and protein in both the MHb and LHb in both. In rat, 39 {+/-} 3% (MHb) and 21 {+/-} 4% (LHb) of cells expressed Oprm1. These proportions were greater in mouse: 57 {+/-} 1% (MHb) and 32 {+/-} 4% (LHb). Within each species, Oprm1 labeling density per positive cell was greater in MHb compared to LHb (p < 0.0001 for rat and mouse). The highest intensity labeling was localized along the lateral edge of the MHb for both methods. ICC showed MOR localized to fibers and somata in MHb and LHb. In LHb, MOR labeling was most dense in intermediate sections along the anterior-posterior (AP) axis. In rats we also observed higher density labeling in dorsal LHb at intermediate AP levels and medial LHb more posteriorly. These results indicate that both MHb and LHb can contribute to MOR mediated actions through their respective circuits.

Key PointsO_LIMu opioid receptor mRNA and protein is expressed in both the medial and lateral habenulae in rat and mouse.
C_LIO_LIIn the medial habenula, most mu opioid receptor mRNA and protein was detected along its lateral border.
C_LIO_LIAcross samples, Oprm1+ cells in the MHb contained more mRNA puncta per cell compared to lateral habenula cells.
C_LI
]]></description>
<dc:creator>Popat, A. K.</dc:creator>
<dc:creator>Simon, R. C.</dc:creator>
<dc:creator>Aoyama, B. B.</dc:creator>
<dc:creator>Wokhlu, A.</dc:creator>
<dc:creator>Ehrlich, A. T.</dc:creator>
<dc:creator>Harwell, C. C.</dc:creator>
<dc:creator>Margolis, E. B.</dc:creator>
<dc:date>2025-12-19</dc:date>
<dc:identifier>doi:10.64898/2025.12.16.694766</dc:identifier>
<dc:title><![CDATA[Mu opioid receptor mRNA and protein localization across the rat and mouse habenula]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.19.695550v1?rss=1">
<title>
<![CDATA[
High false sign rates in transcriptome-wide association studies 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.19.695550v1?rss=1</link>
<description><![CDATA[
Transcriptome-wide association studies (TWAS) are widely used to identify genes involved in complex traits and to infer the direction of gene effects on traits. However, despite their popularity, it remains unclear how accurately TWAS recover the true direction of a genes effect on a trait. Here, we estimate the false sign rate (FSR) of TWAS for plasma proteins, leveraging the expectation that increased gene expression should generally increase protein expression. We then extend this framework to complex traits, where loss-of-function burden tests provide the expected direction-of-effect. In both analyses, we observe high discordance with expectations, with TWAS showing an FSR of 23% for plasma proteins and 33% for complex traits. While colocalization-based filtering reduced the FSR, substantial discordance remained, and with substantial loss of recall. However, when we restricted gene-direction assignments for plasma proteins to using only relevant tissues in combination with colocalization-based filtering, the FSR dropped to 11%, and to just 5% if we excluded brain-specific proteins. We propose that much of the sign discordance arises when eQTLs in non-trait-relevant tissues tag GWAS-associated haplotypes via distinct, tightly-linked regulatory variants, yielding spurious TWAS associations with the correct genes but with unreliable direction-of-effect. These findings show that TWAS-based direction-of-effect estimates should be interpreted with caution and raise concerns about the reliability of TWAS more broadly.
]]></description>
<dc:creator>Gerlach, P. A.</dc:creator>
<dc:creator>Milind, N.</dc:creator>
<dc:creator>Spence, J. P.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:date>2025-12-20</dc:date>
<dc:identifier>doi:10.64898/2025.12.19.695550</dc:identifier>
<dc:title><![CDATA[High false sign rates in transcriptome-wide association studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.18.695199v1?rss=1">
<title>
<![CDATA[
Single cell RNA sequencing reveals limited effects of Lrrk2 genotype in a mouse model of acute CNS inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.18.695199v1?rss=1</link>
<description><![CDATA[
BACKGROUNDPrevious data implicates neuroinflammation in the pathogenesis of Parkinsons disease (PD). Of the genes associated with PD, Leucine rich repeat kinase 2 (LRRK2) has previously been proposed to play a role in neuroinflammation. However, the extent to which LRRK2 is involved in endogenous inflammatory signaling is unclear.

OBJECTIVETo examine whether endogenous mutations in LRRK2 affect responses to neuroinflammation in vivo.

METHODSWe injected cohorts of mice with knock-in mutations in Lrrk2, homologous to those causing human PD, with a single intrastriatal injection of lipopolysaccharide (LPS) or control. We used single cell RNA-Sequencing to examine cell type specific responses to treatment and genotype and validated key results with orthogonal approaches.

RESULTSWe found that our chosen paradigm of acute LPS exposure evokes robust transcriptional changes consistent with a multicellular neuroinflammatory response. We also found evidence of peripheral immune cell recruitment into the brain and interaction with brain-resident microglia. However, the transcriptional effects of Lrrk2 mutations were limited to small numbers of genes, including down regulation of gene ontogeny terms related to lysosomes, in microglia.

CONCLUSIONSOur data clearly demonstrate that many cells in the brain respond to a single inflammatory insult with strong transcriptional responses and that, even in a model focused on CNS injection, there is interaction between peripheral and central immune cells. In contrast, the quantitative effects of Lrrk2 mutations are modest at the transcriptional level, demonstrating that additional studies are needed to clarify whether Lrrk2 mutations affect neuroinflammation in an endogenous context.
]]></description>
<dc:creator>Cookson, M. R.</dc:creator>
<dc:creator>Kaganovich, A.</dc:creator>
<dc:creator>Horan-Portelance, L.</dc:creator>
<dc:creator>Landeck, N.</dc:creator>
<dc:creator>Acri, D. J.</dc:creator>
<dc:creator>Ding, J.</dc:creator>
<dc:creator>Langston, R.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.18.695199</dc:identifier>
<dc:title><![CDATA[Single cell RNA sequencing reveals limited effects of Lrrk2 genotype in a mouse model of acute CNS inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.18.695250v1?rss=1">
<title>
<![CDATA[
Neocortical temporal patterning by a two-layered regulatory network 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.18.695250v1?rss=1</link>
<description><![CDATA[
In the developing neocortex, a diverse array of neurons with defined types and abundances are systematically generated by a limited population of radial glial progenitors (RGPs) as they undergo successive fate changes. The molecular regulation behind this intricate temporal patterning remains elusive. We undertook in-depth single-cell multi-omics analyses, discovering a two-layered regulatory framework at the core of this process. Central to this are global temporal regulators positioned above a temporal network, consisting of series of transcriptional factor (TF) hub groups under sequential state transitions. This temporal network operates not by restricting TF expressions to discrete temporal windows, but through coordinated transcriptional and chromatin-accessibility dynamics that modulate transient TF regulatory activity. Moreover, global temporal regulators specify the duration of each cascading stage and, consequently, the number of progenies generated at each stage. Loss of global temporal regulators protracts RGP lineage progression, whereas their increased activity accelerates it. These findings suggest a two-layer temporal regulatory system controlling RGP lineage progression and neural progeny output duality in mammalian neocortical development.
]]></description>
<dc:creator>Yuan, G.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Wang, X.-W.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Yu, X.</dc:creator>
<dc:creator>Zhu, M.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Zhang, N.</dc:creator>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Dai, L.</dc:creator>
<dc:creator>Liu, Y.-Y.</dc:creator>
<dc:creator>Shi, S.-H.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.18.695250</dc:identifier>
<dc:title><![CDATA[Neocortical temporal patterning by a two-layered regulatory network]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.19.695001v1?rss=1">
<title>
<![CDATA[
Transcriptomic analysis of 40 human and rodent skeletal muscle exerkines 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.19.695001v1?rss=1</link>
<description><![CDATA[
Animal and human studies show that exercise induces organism wide molecular adaptations, many of which are mediated by exerkines which are secreted factors that enable communication between tissues such as skeletal muscle, adipose tissue, liver and the brain. However, the tissue specific responsiveness of individual exerkines and how these responses differ across species, exercise conditions and sexes remain poorly understood. To address this gap, we systematically analyzed skeletal muscle transcriptomic responses of 40 exerkines using three publicly available datasets which include MetaMEx, Extrameta and the MoTrPAC rat training study. We analyzed exerkine specific regulation in humans, mice and rats across acute and chronic exercise as well as inactivity, and determined which responses were conserved, species specific, sex dependent or dependent on exercise duration. Our analysis reveals substantial heterogeneity in skeletal muscle exerkine regulation with only a small subset showing conserved changes across species, while many exerkines exhibited human exclusive, rodent exclusive, acute specific or chronic specific patterns. These results provide a ranked overview of the most exercise responsive skeletal muscle exerkines and highlight the need for multi species and multi condition approaches when selecting exerkines as biomarkers or therapeutic targets.
]]></description>
<dc:creator>Taha, H. B.</dc:creator>
<dc:creator>Robbins, N.</dc:creator>
<dc:creator>Zoha, F.-S.</dc:creator>
<dc:creator>Vivek, N.</dc:creator>
<dc:creator>Bogoniewski, A.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.19.695001</dc:identifier>
<dc:title><![CDATA[Transcriptomic analysis of 40 human and rodent skeletal muscle exerkines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.19.695538v1?rss=1">
<title>
<![CDATA[
Ecdysone Receptor autonomously controls germ cell differentiation in the Drosophila ovary 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.19.695538v1?rss=1</link>
<description><![CDATA[
In the Drosophila ovary, the steroid hormone ecdysone controls germline stem cell (GSC) maintenance and germ cell differentiation. Prior studies demonstrated that ecdysone regulates germ cell differentiation non-autonomously via the nuclear receptor Ecdysone Receptor (EcR) in ovarian somatic cells. Although EcR is also expressed in GSCs and their differentiating daughters, potential direct roles for EcR in GSCs independent of the soma have not been examined. Here, we demonstrate that EcR functions autonomously in GSCs and cystoblasts to control germline differentiation. While depletion of EcR from GSCs mildly reduces GSC self-renewal, over-expression of EcR specifically in GSCs and cystoblasts impedes germ cell differentiation, phenotypically resembling bag of marbles loss-of-function and Bone Morphogenetic Protein signaling constitutive activation. We propose that while low levels of EcR are essential to maintain GSC self-renewal and permit initial differentiation, higher levels of EcR accumulate in differentiated germ cells to promote transcription of maternal genes, providing temporal control over germline differentiation. These data support the model that stem cells harbor unique mechanisms to integrate signals from multiple cell sources that safeguard their self-renewal in response to local and physiological cues.

SUMMARY STATEMENTThe nuclear receptor EcR modulates stem cell maintenance and differentiation in ovarian germ cells.
]]></description>
<dc:creator>Jung, L. E.</dc:creator>
<dc:creator>Warren, A. I.</dc:creator>
<dc:creator>Yin, C.</dc:creator>
<dc:creator>Huang, W.</dc:creator>
<dc:creator>Simmons, A. C.</dc:creator>
<dc:creator>McDonald, S. I.</dc:creator>
<dc:creator>Swain, L. A.</dc:creator>
<dc:creator>Garrido, V. E.</dc:creator>
<dc:creator>Phipps, D. N.</dc:creator>
<dc:creator>Cesar, B.</dc:creator>
<dc:creator>Finger, D. S.</dc:creator>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Nystul, T.</dc:creator>
<dc:creator>Ables, E. T.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.19.695538</dc:identifier>
<dc:title><![CDATA[Ecdysone Receptor autonomously controls germ cell differentiation in the Drosophila ovary]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.23.696243v1?rss=1">
<title>
<![CDATA[
Concreteness shapes semantic representations in bilingual brains 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.23.696243v1?rss=1</link>
<description><![CDATA[
Behavioral studies show that humans process concrete words more similarly across languages than abstract words. This suggests that in bilingual brains, semantic representations may be more similar across languages for concrete concepts than for abstract concepts, but existing neuroimaging evidence is inconclusive. Here, we analyzed functional magnetic resonance imaging (fMRI) data from fluent Chinese-English bilinguals who read several hours of naturalistic narratives in both languages. We used encoding models to estimate voxelwise tuning towards concrete and abstract concepts in each language separately. We then quantified the similarity of cortical semantic representations across languages. First, we find that the cortical organization of concreteness tuning is consistent across languages. Second, semantic representations are similar across languages for both concrete-tuned voxels and abstract-tuned voxels. Third, we find that for abstract concepts, cross-language similarity of semantic representations may be driven by emotionality. Overall, these findings reveal how concreteness affects semantic representations in bilingual brains.
]]></description>
<dc:creator>Lamarre, M.</dc:creator>
<dc:creator>Gong, X. L.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Deniz, F.</dc:creator>
<dc:date>2025-12-24</dc:date>
<dc:identifier>doi:10.64898/2025.12.23.696243</dc:identifier>
<dc:title><![CDATA[Concreteness shapes semantic representations in bilingual brains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.23.696273v1?rss=1">
<title>
<![CDATA[
Genome-scale perturb-seq in primary human CD4+ T cells maps context-specific regulators of T cell programs and human immune traits 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.23.696273v1?rss=1</link>
<description><![CDATA[
Gene regulatory networks encode the fundamental logic of cellular functions, but systematic network mapping remains challenging, especially in cell states relevant to human biology and disease. Here, we perturbed all expressed genes across 22 million primary human CD4+ T cells from four donors and developed a probe-based perturb-seq platform to measure the transcriptome effects in cells at rest and after stimulation. These data allow us to map genes that regulate known and novel pathways, including novel regulators of cytokine production. Importantly, active regulators and the gene programs they control change dramatically across stimulation conditions. Perturbation signatures enabled us to model T cell states observed in population-scale transcriptomic atlases, nominating regulators of Th1 and Th2 polarization and of age-related T cell phenotypes. Finally, we leveraged perturb-seq to implicate context-specific gene regulatory pathways in autoimmune disease risk. Our data provide a foundational resource to decode human immune function and genetic variation and for new approaches to study gene regulatory networks.
]]></description>
<dc:creator>Zhu, R.</dc:creator>
<dc:creator>Dann, E.</dc:creator>
<dc:creator>Yan, J.</dc:creator>
<dc:creator>Reyes Retana, J.</dc:creator>
<dc:creator>Goto, R.</dc:creator>
<dc:creator>Guitche, R. C.</dc:creator>
<dc:creator>Petersen, L. K.</dc:creator>
<dc:creator>Ota, M.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:date>2025-12-24</dc:date>
<dc:identifier>doi:10.64898/2025.12.23.696273</dc:identifier>
<dc:title><![CDATA[Genome-scale perturb-seq in primary human CD4+ T cells maps context-specific regulators of T cell programs and human immune traits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.23.696294v1?rss=1">
<title>
<![CDATA[
Shifts in the human gut microbiome during cancer chemotherapy are diet-dependent 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.23.696294v1?rss=1</link>
<description><![CDATA[
Numerous studies have implicated both dietary intake and the human gut microbiome in colorectal cancer (CRC) treatment outcomes. However, little is known about how patients adjust their dietary intake during cancer chemotherapy or if these dietary changes contribute to treatment-associated alterations in the gut microbiome. We performed paired longitudinal diet and microbiome analysis during CRC treatment with oral fluoropyrimidines (NCT04054908) and validated key associations using cell culture assays. Diet quality significantly decreased during chemotherapy. Carbohydrate and refined grain intake increased, accompanied by decreased consumption of fats, nuts and seeds, and fat-soluble micronutrients. Multiple individual dietary components were strongly linked to the gut microbiome. Decreases in theobromine intake correlated with decreases in overall microbial diversity and more gastrointestinal toxicities. Diet shifts partly explained changes in bacterial abundance during chemotherapy, including more severe depletion of Faecalibacterium prausnitzii in patients with decreased vitamin K1 intake. Changes in diet were correlated with multiple bacterial gene families involved in micronutrient metabolism and drug sensitivity. Increased copper intake was linked to decreased Fusobacterium nucleatum in patients and inhibited F. nucleatum in cell assays. Together, these data suggest that chemotherapy-related decreases in diet quality and micronutrient intake contribute to changes in gut bacterial diversity, taxonomic composition, and gene abundance. Our approach may generalize to other cancer therapies and emphasizes the need for collecting more robust dietary data in clinical microbiome studies.
]]></description>
<dc:creator>Trepka, K. R.</dc:creator>
<dc:creator>Van Blarigan, E. L.</dc:creator>
<dc:creator>Kyaw, T. S.</dc:creator>
<dc:creator>Olson, C. A.</dc:creator>
<dc:creator>Partipilo, G.</dc:creator>
<dc:creator>Ortega, E. F.</dc:creator>
<dc:creator>Noecker, C.</dc:creator>
<dc:creator>Upadhyay, V.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Gempis, D.</dc:creator>
<dc:creator>Steiding, P.</dc:creator>
<dc:creator>Stanfield, D.</dc:creator>
<dc:creator>Venook, A. P.</dc:creator>
<dc:creator>Atreya, C. E.</dc:creator>
<dc:creator>Kidder, W. A.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:date>2025-12-24</dc:date>
<dc:identifier>doi:10.64898/2025.12.23.696294</dc:identifier>
<dc:title><![CDATA[Shifts in the human gut microbiome during cancer chemotherapy are diet-dependent]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.23.696255v1?rss=1">
<title>
<![CDATA[
Mammalian metaphase kinetochores are elastic and require condensin for robust structure and function 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.23.696255v1?rss=1</link>
<description><![CDATA[
SummaryTran et al. show that mammalian metaphase kinetochores undergo large-scale, complex, and elastic deformations under forces of different timescales, magnitudes and directions. They demonstrate that condensin is required for kinetochore local structural organization and microtubule attachment under force.

AbstractThe mammalian kinetochore connects chromosomes to dynamic spindle microtubules. To remain attached, it must maintain its structural integrity under force, but to what extent and how it does so remain unclear. Under spindle forces, we find using super resolution microscopy that inner (CENP-A) and outer (Hec1) metaphase kinetochores undergo correlated, large-scale (1 {micro}m) deformations along the force axis, suggesting dynamic, relative sliding of parallel protein linkages. Kinetochore shape changes can be asymmetric, with centromere-facing "tails" correlating with erroneous attachment geometries. Applying microneedle pulling forces, we demonstrate that kinetochores are elastic, stretching under force and relaxing in seconds afterwards. Finally, we show that SMC2 depletion results in more variable kinetochore deformations, despite maintained elasticity, and in reduced microtubule attachment stability. Thus, the kinetochore is structurally highly dynamic and requires a stable centromere base to maintain its structure and function under force. We propose a model whereby individual protein linkages are stiff yet global kinetochore structure flexible to accommodate different attachment geometries and forces while maintaining function.
]]></description>
<dc:creator>Tran, V. M.</dc:creator>
<dc:creator>Tao, J.</dc:creator>
<dc:creator>Rux, C. J.</dc:creator>
<dc:creator>Gomez Siu, D.</dc:creator>
<dc:creator>Rosas Salvans, M.</dc:creator>
<dc:creator>Dumont, S.</dc:creator>
<dc:date>2025-12-25</dc:date>
<dc:identifier>doi:10.64898/2025.12.23.696255</dc:identifier>
<dc:title><![CDATA[Mammalian metaphase kinetochores are elastic and require condensin for robust structure and function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.23.695206v1?rss=1">
<title>
<![CDATA[
Aging increases ovarian cancer growth, metastasis and immunosuppression that can be alleviated by inhibiting hedgehog signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.23.695206v1?rss=1</link>
<description><![CDATA[
Ovarian cancer incidence and mortality increase with age, yet the impact of physiological aging on tumor progression and the tumor immune microenvironment remains poorly defined. Here we show using orthotopic implantation of two syngeneic models representing distinct cellular origins (ovarian surface epithelial and fallopian tube-derived) in young versus aged mice, that aged hosts exhibit markedly higher tumor burden, ascites accumulation, and proliferation. Selective follicle depletion in young mice using VCD did not recapitulate these effects, indicating that age-associated microenvironmental changes beyond hormonal decline drive tumor growth. Spatial transcriptomics revealed distinct intratumoral heterogeneity in both age groups. Comparison of CD45+ cells between aged and young tumors showed Hedgehog signaling enrichment and immunosuppressive signatures in aged hosts, with elevated M2 macrophages and Foxp3+ regulatory T cells. Notably, pharmacologic inhibition of Hedgehog signaling with vismodegib in aged mice suppressed tumor growth, reduced metastatic spread, and decreased infiltration of CD206+ macrophages and Foxp3+ T cells while sparing CD8+ T cells. Our findings provide proof-of-concept that vismodegib can reduce tumor growth and specific immunosuppressive populations in aged hosts, suggesting Hedgehog inhibition as a potential immunomodulatory strategy for older ovarian cancer patients or those tumors that exhibit Hedgehog pathway activation.
]]></description>
<dc:creator>Kumari, A.</dc:creator>
<dc:creator>Elbahoty, M. H.</dc:creator>
<dc:creator>Rajkarnikar, R.</dc:creator>
<dc:creator>Sureja, K.</dc:creator>
<dc:creator>Monavarian, M.</dc:creator>
<dc:creator>Quintero, L. M.</dc:creator>
<dc:creator>Fuh, K.</dc:creator>
<dc:creator>Tyrrell, D. J.</dc:creator>
<dc:creator>Shevde, L. A.</dc:creator>
<dc:creator>Margaroli, C.</dc:creator>
<dc:creator>Mythreye, K.</dc:creator>
<dc:date>2025-12-27</dc:date>
<dc:identifier>doi:10.64898/2025.12.23.695206</dc:identifier>
<dc:title><![CDATA[Aging increases ovarian cancer growth, metastasis and immunosuppression that can be alleviated by inhibiting hedgehog signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.27.696675v1?rss=1">
<title>
<![CDATA[
Dose-dependent NFI regulation of progenitor lifespan and output underlying human neocortical malformation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.27.696675v1?rss=1</link>
<description><![CDATA[
Nuclear Factor I (NFI) misexpressions in humans are associated with severe brain malformations, yet the underlying mechanisms remain poorly understood. Here we show that NFIs regulate the broad lineage progression and lifespan of radial glial progenitors (RGPs), and consequently neocortical development in a dose-dependent manner. Human cerebral organoids carrying patient-mimicking NFI mutations exhibit expression level-dependent bidirectional impairments in RGP temporal development, coinciding with patient phenotypes. In mouse models, selective removal of NFIs leads to a dramatic protraction of RGP lineage progression and lifespan, excessive progeny output, and neocortical overgrowth and abnormal folding, whereas overexpression of NFIs accelerates RGP lineage progression, resulting in developmental stage-dependent precocious productions of diverse neural progenies. Moreover, NFIs exhibit a positive auto-regulation and progressive increase in expression, and regulate distinct temporal-specific targets underlying RGP lineage progression. These results suggest that NFIs act as evolutionarily conserved key global temporal regulators of RGP lineage progression and neocortical development.

HIGHLIGHTSO_LINFIs affect human RGP temporal development coinciding with patient phenotypes.
C_LIO_LINFI removal protracts RGP lineage progression and lifespan with excessive progeny output.
C_LIO_LINFI overexpression accelerates RGP lineage progression with precocious progeny output.
C_LIO_LINFIs regulate distinct temporal-specific targets underlying RGP lineage progression.
C_LI
]]></description>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Yuan, G.</dc:creator>
<dc:creator>Albizzati, E.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Yu, X.</dc:creator>
<dc:creator>Chang, X.</dc:creator>
<dc:creator>Lee, C. H.</dc:creator>
<dc:creator>Du, H.</dc:creator>
<dc:creator>Lao, Z.</dc:creator>
<dc:creator>Krishnamurthy, A.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Lv, X.</dc:creator>
<dc:creator>Tang, X.</dc:creator>
<dc:creator>Hu, S.</dc:creator>
<dc:creator>Chi, Y.</dc:creator>
<dc:creator>Ma, J.</dc:creator>
<dc:creator>Gronostajski, R. M.</dc:creator>
<dc:creator>Richards, L. J.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Joyner, A. L.</dc:creator>
<dc:creator>Tchieu, J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Shi, S.-H.</dc:creator>
<dc:date>2025-12-27</dc:date>
<dc:identifier>doi:10.64898/2025.12.27.696675</dc:identifier>
<dc:title><![CDATA[Dose-dependent NFI regulation of progenitor lifespan and output underlying human neocortical malformation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.25.696505v1?rss=1">
<title>
<![CDATA[
Large scale prospective evaluation of co-folding across 557 Mac1-ligand complexes and three virtual screens 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.25.696505v1?rss=1</link>
<description><![CDATA[
Accurate prediction of ligand-bound protein complexes and ranking them by affinity are central problems in drug discovery. While deep learning co-folding methods can help address these challenges, their evaluation has been hampered by the difficulties in assessing independence from training data and insufficiently large test sets. Here we test the ability of co-folding methods to predict the structures of 557 ligands bound to the SARS-CoV-2 NSP3 macrodomain (Mac1) that were determined after the training cut-off dates. AlphaFold3 (AF3), Boltz-2, and Chai-1 each reproduced >50% of the Mac1 ligand poses to better than 2 [A] RMSD of experiment. Despite the potential for co-folding to describe protein conformational changes that stabilize ligand binding, we did not find that common conformational rearrangements, including peptide flip and a large loop opening, were recapitulated by the co-folding prediction. For AF3 and Chai-1, ligand pose prediction confidence weakly, but significantly, tracked experimental potency, while DOCK3.7 energies were only weakly correlated. Boltz-2 affinity predictions showed the strongest correlation with measured potency and, after calibration, achieved lower mean absolute error than a baseline predictor. We next assessed whether co-folding scores could rescore docking hit-lists to distinguish true ligands from non-binders among hundreds of molecules prospectively experimentally tested against AmpC {beta}-lactamase, the dopamine D4 and the {sigma}2 receptors. AF3 ligand pose confidence values did not separate true ligands from high-scoring false-positives as effectively as docking scores or Boltz-2 affinity predictions did. Taken together, the modest, but independent correlations of docking score and co-folding confidence or affinity suggests that integrating physics-based and deep-learning and approaches may help with hit prioritization and subsequent optimization in structure-based ligand discovery.
]]></description>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Correy, G. J.</dc:creator>
<dc:creator>Hall, B. W.</dc:creator>
<dc:creator>Rachman, M. M.</dc:creator>
<dc:creator>Mailhot, O.</dc:creator>
<dc:creator>Togo, T.</dc:creator>
<dc:creator>Gonciarz, R. L.</dc:creator>
<dc:creator>Jaishankar, P.</dc:creator>
<dc:creator>Neitz, R. J.</dc:creator>
<dc:creator>Hantz, E. R.</dc:creator>
<dc:creator>Doruk, Y. U.</dc:creator>
<dc:creator>Stevens, M. G. V.</dc:creator>
<dc:creator>Diolaiti, M. E.</dc:creator>
<dc:creator>Reid, R.</dc:creator>
<dc:creator>Gopalkrishnan, S.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Renslo, A. R.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:date>2025-12-28</dc:date>
<dc:identifier>doi:10.64898/2025.12.25.696505</dc:identifier>
<dc:title><![CDATA[Large scale prospective evaluation of co-folding across 557 Mac1-ligand complexes and three virtual screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.29.696818v1?rss=1">
<title>
<![CDATA[
Sleep-Wake Transitions Are Impaired in the AppNL-G-FMouse Model of Early Onset Alzheimers Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.29.696818v1?rss=1</link>
<description><![CDATA[
Poor sleep quality and reduced sleep duration are associated with Alzheimers disease (AD)-related {beta}-amyloid (A{beta}) pathologies. We conducted two studies of sleep/wake, activity and body temperature in AppNL-G-F mice, a strain that exhibits three mutations in the human App gene associated with elevated risk for early onset AD. First, AppNL-G-F mice were compared to wildtype (WT) littermates at 14-18 and 18-22 months of age and, at both ages, were found to exhibit more Wake and less NREM and REM sleep than WT littermates. This long wake/short sleep phenotype was evident during the dark phase at 14-18 months but occurred in both the light and dark phases at 18-22 months. AppNL-G-F mice had fewer short (<60 sec) and more long (>260 sec) Wake bouts and were hyperactive at 18-22 months, which undoubtedly contributed to the increased Wake/reduced sleep. Despite this reduced sleep phenotype, AppNL-G-F mice were no sleepier than WT mice and the sleep homeostat was functional in both strains. In the second study, sex differences in these parameters were assessed at 18-24 months. Reduced sleep was evident in both sexes of AppNL-G-F mice but was clearly more evident in females. Wake and REM sleep bout durations were longer in both sexes of AppNL-G-F mice than in WT littermates. EEG spectral power during NREM sleep was reduced in female AppNL-G-F mice between 4.88-10.50 Hz compared to WT mice whereas, during REM sleep, both male and female AppNL-G-F mice exhibited reduced spectral power in the theta range. These results suggest that A{beta} deposition may impair state transition mechanism(s) in AppNL-G-F mice and demonstrate that, as in human AD patients, the long wake/short sleep phenotype was more evident in female AppNL-G-F mice, thus supporting the use of this strain as a model to investigate interventions that mitigate AD burden during early disease stages.
]]></description>
<dc:creator>Kilduff, T. S.</dc:creator>
<dc:creator>Tisdale, R. K.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Miller, S. R.</dc:creator>
<dc:creator>Lee, S. M.</dc:creator>
<dc:creator>Park, S.</dc:creator>
<dc:creator>Shin, J.</dc:creator>
<dc:creator>Allocca, G.</dc:creator>
<dc:creator>Palop, J.</dc:creator>
<dc:date>2025-12-29</dc:date>
<dc:identifier>doi:10.64898/2025.12.29.696818</dc:identifier>
<dc:title><![CDATA[Sleep-Wake Transitions Are Impaired in the AppNL-G-FMouse Model of Early Onset Alzheimers Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.29.696958v1?rss=1">
<title>
<![CDATA[
Type 2 diabetes Reprograms Bone Marrow Hematopoiesis and Dysregulates Immune Signaling in Response to Stroke 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.29.696958v1?rss=1</link>
<description><![CDATA[
Type 2 diabetes mellitus (T2DM) worsens stroke outcomes, but the underlying mechanisms linking T2DM to systemic immune dysfunction remain unclear. We investigated whether T2DM alters bone marrow (BM) hematopoiesis and dysregulate immune signaling following ischemic stroke in mice. Single-cell RNA sequencing, GeoMx digital spatial profiling (DSP), nCounter, and flow cytometry were used to analyze BM cells from control (db/+, Ctrl) and diabetic (db/db, T2DM) mice underwent experimental stroke or sham surgery. Diabetes caused marked structural remodeling of BM, with reduced cellularity and imbalance of hematopoietic lineages. Pseudotime trajectory analysis revealed impaired differentiation and maturation signatures of hematopoietic progenitor cells (HPC1) and granulocytes, and overactivation toward monocytes in diabetes after stroke. CellChat analysis demonstrated reorganization of intercellular communication, with hematopoietic progenitor cells (HPC1) and monocytes emerging as dominant signaling hubs through upregulated MIF, SIRP, and THBS pathways. AUCell enrichment indicated increased glycolysis and oxidative phosphorylation but reduced interferon-{gamma} (IFN-{gamma}) signaling, reflecting metabolic activation coupled with immune dysregulation. DSP and nCounter further confirmed upregulation of genes in the MIF, SIRP and THBS pathways in CD115 monocytes and Ly6G neutrophils, indicating proinflammatory and migratory activation in diabetic bone marrow. Our data suggest that T2DM reprograms hematopoiesis and signaling networks, leading to maladaptive myeloid responses and impaired immune regulation after stroke. This maladaptive BM environment amplifies inflammation and limits repair, linking diabetic metabolic stress to worsened ischemic outcomes. Targeting bone marrow immune dysfunction may offer a therapeutic strategy to improve stroke recovery in diabetic patients.
]]></description>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Gu, W.</dc:creator>
<dc:creator>Yu, K.</dc:creator>
<dc:creator>Tu, C.-L.</dc:creator>
<dc:creator>Chang, W.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:date>2025-12-30</dc:date>
<dc:identifier>doi:10.64898/2025.12.29.696958</dc:identifier>
<dc:title><![CDATA[Type 2 diabetes Reprograms Bone Marrow Hematopoiesis and Dysregulates Immune Signaling in Response to Stroke]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.30.697040v1?rss=1">
<title>
<![CDATA[
The osm-9 mutant strain is defective in the recovery phase of long-term memory dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.30.697040v1?rss=1</link>
<description><![CDATA[
Memory by definition is the storage of learned information from experiences where it can be recalled in the future. Memory is essential for organisms of any level, from single celled bacteria to complex mammals like whales in order for them to adapt and survive. The storage of long term memory is known to start in the hippocampus but long-term memory is not simply contained in this one region in the brain but across multiple regions including the cortex, amygdala and nucleus accumbens. This diversity of locations that memory can be located in prompts the question of what molecular components within the nervous system are required to establish and maintain long term memory. To dive into this, we utilize the simple neuroanatomy of C. elegans with only 302 neurons to examine the effects of mutations of specific genes on the success of forming long term memories. Previous work established that C. elegans with defective OSM-9 proteins are unable to sustain an aversion to butanone (an odor that they are innately attracted to) after it is paired once with a negative stimulus. This helped established the importance of OSM-9 proteins for short term memory but how can the OSM-9 protein - a homolog to human TRPV5 and TRPV6 commonly associated with recognising pain and thermoregulation rather than memory - play a part within the dynamics of space training induced long term memory? Here we show that the loss of the OSM-9 protein affects the dynamics of memory consolidation, which is the process by which short term memory is converted to long term memory. Previously, OSM-9 protein was implicated in the formation of short term memory which was termed adaptation. Here, we examine spaced training induced memory that is resistant to re-feeding and depends on sleep. This study indicates that after spaced training, the OSM-9 protein is required for the conversion of short term memory to long term memory and not acquisition. Furthermore, we confirmed that 30 minutes on food after spaced training, Wild type C. elegans seem to lose their memory but gain it back 120 minutes after, emphasizing the dynamics of memory and revealing that it is not a smoothly continuous process. Our results demonstrate that TRPV containing channels plays a role in stabilizing memory for long term use. Analyzing the specific point in which the OSM-9 protein plays a part in reactivating memory may help pave the way to a more detailed understanding of memory dynamics and the processes involved in recovering memory. Future work includes: identifying where the loss of memory goes, whether its masked or truly disappears can further explain what the OSM-9 protein supports when long term memory is formed; understanding if OSM-9 is directly involved with the process of "reviving" the memory or is it involved in the development of the C. elegans circuit which allows for this process to occur.
]]></description>
<dc:creator>Ma, N. I.</dc:creator>
<dc:creator>Saifuddin, M. F. A.</dc:creator>
<dc:creator>Munoz Lobato, F.</dc:creator>
<dc:creator>L'Etoile, N. D.</dc:creator>
<dc:date>2025-12-30</dc:date>
<dc:identifier>doi:10.64898/2025.12.30.697040</dc:identifier>
<dc:title><![CDATA[The osm-9 mutant strain is defective in the recovery phase of long-term memory dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.30.697009v1?rss=1">
<title>
<![CDATA[
Kinematic fields of the visuo-hippocampal circuit 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.30.697009v1?rss=1</link>
<description><![CDATA[
The structure of focused receptive fields in retinotopic and allocentric space in the visual cortex and hippocampus is studied extensively to understand cognitive functions such as stimulus tracking and path-integration, respectively. These functions require accurate tracking of running speed and acceleration. While speed dependence in these areas is well-studied, the acceleration dependence has received little attention, and the joint effect of speed-acceleration is unknown. Hence, we assessed the joint influence of these kinematic variables by computing receptive fields in a novel, 2D phase space of speed-acceleration using the Allen Brain Visual Observatory data. Head-fixed mice ran spontaneously on a running wheel next to a monocular gray screen ensuring virtually no changes in the retinotopic or allocentric space. Remarkably, about half (44.3%, 6590/14862) of neurons in the visuo-hippocampal circuit (from LGN to subiculum) were significantly modulated by a specific combination of speed and acceleration to form receptive fields in the speed-acceleration phase space (called kinematic space). The prevalence of kinematic tuning is comparable to the fraction of place cells in this part of the mouse hippocampus, and the fraction of visually tuned neurons in higher visual areas. The kinematic field size ([~]30% of the sampled space) was also comparable to the place field size. Kinematic fields spanned the entire space but preferred either low-speed, high-acceleration or the high-speed, low-acceleration segments. Although pupil size too varied spontaneously, the kinematic tuning exceeded pupil size tuning in all brain areas. Surprisingly, pupil size modulated the activity of [~]20% of hippocampal neurons, which was comparable to that in LGN and visual cortical areas. Thus, all visuo-hippocampal areas have focused kinematic fields that are three times more prevalent than the pupil size fields in the absence of any vestibular, visual or spatial stimuli. This kinematic phase space could interface with retinotopic and allocentric spaces to guide natural behaviors involving movement.
]]></description>
<dc:creator>Purandare, C.</dc:creator>
<dc:creator>mehta, m. R.</dc:creator>
<dc:date>2025-12-31</dc:date>
<dc:identifier>doi:10.64898/2025.12.30.697009</dc:identifier>
<dc:title><![CDATA[Kinematic fields of the visuo-hippocampal circuit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.30.697051v1?rss=1">
<title>
<![CDATA[
Multi-modal choroid plexus pathology in aging and Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.30.697051v1?rss=1</link>
<description><![CDATA[
Brain barriers, cerebrospinal fluid (CSF) dynamics, and peripheral factors are implicated as significant contributors to Alzheimers disease (AD). The choroid plexus (ChP) is a blood-brain interface that produces CSF and forms the blood-CSF barrier. However, how ChP pathology develops across the lifespan and contributes to AD has not been systematically characterized. Here, we report a multi-modal ChP atlas integrating single-nucleus transcriptomics from 49 individuals, AI-assisted quantitative histopathology across >500 postmortem samples age 16 to 105, spatial transcriptomics, and functional studies in 5xFAD mice. We identify fibrosis, calcification, and macrophage abnormalities as hallmarks of ChP aging, with AD pathology conferring additional effects, including expansion of a pro-inflammatory fibroblast-macrophage signaling niche. In 5xFAD mice, macrophage dysfunction is associated with impaired epithelial barrier maintenance and repair. Together, these data provide a foundational resource for understanding ChP dysfunction in aging and AD and propose the macrophage-fibroblast-epithelial barrier axis as a driver of ChP pathology.
]]></description>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Lotfy, P.</dc:creator>
<dc:creator>Englert, B.</dc:creator>
<dc:creator>Oberhauser, J.</dc:creator>
<dc:creator>Byer, L. I. J.</dc:creator>
<dc:creator>Wihlman, J.</dc:creator>
<dc:creator>Colangelo, K.</dc:creator>
<dc:creator>Okar, S. V.</dc:creator>
<dc:creator>Thommana, A.</dc:creator>
<dc:creator>Puttonen, H.</dc:creator>
<dc:creator>Mäyränpää, M. I.</dc:creator>
<dc:creator>Tuimala, J.</dc:creator>
<dc:creator>Pedrosa, R.</dc:creator>
<dc:creator>Kumar, D.</dc:creator>
<dc:creator>Haberberger, J. F.</dc:creator>
<dc:creator>Atkins, M. E.</dc:creator>
<dc:creator>Alimukhamedov, S.</dc:creator>
<dc:creator>Pragana, A.</dc:creator>
<dc:creator>Benson, J.</dc:creator>
<dc:creator>Gabrielle, M. E.</dc:creator>
<dc:creator>Dong, A.</dc:creator>
<dc:creator>DurantLaforet, V.</dc:creator>
<dc:creator>Lin, P. B.-C.</dc:creator>
<dc:creator>Keene, C. D.</dc:creator>
<dc:creator>Latimer, C. S.</dc:creator>
<dc:creator>Prater, K. E.</dc:creator>
<dc:creator>Holtzman, D. M.</dc:creator>
<dc:creator>Isakova, A. E.</dc:creator>
<dc:creator>Wyss-Coray, T.</dc:creator>
<dc:creator>Schafer, D. P.</dc:creator>
<dc:creator>Reich, D. S.</dc:creator>
<dc:creator>Lehtimäki, T.</dc:creator>
<dc:creator>Karhunen, P. J.</dc:creator>
<dc:creator>Kok, E.</dc:creator>
<dc:creator>Jansson, D.</dc:creator>
<dc:creator>Yang, A. C.</dc:creator>
<dc:creator>Myllykangas, L.</dc:creator>
<dc:creator>Ordovas-Montanes, J.</dc:creator>
<dc:creator>Lehtinen, M. K.</dc:creator>
<dc:date>2025-12-31</dc:date>
<dc:identifier>doi:10.64898/2025.12.30.697051</dc:identifier>
<dc:title><![CDATA[Multi-modal choroid plexus pathology in aging and Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.31.697099v1?rss=1">
<title>
<![CDATA[
Fezf2 regulates differentiation of Aire-expressing and post-Aire mimetic epithelial populations maintaining thymic homeostasis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.31.697099v1?rss=1</link>
<description><![CDATA[
Fezf2 has been proposed to serve principally as a transcriptional regulator of broad self-antigen expression in medullary thymic epithelial cells (mTECs). Here, we demonstrate an additional function for Fezf2 as a developmental regulator of TEC lineage commitment. Fezf2 deficiency in the thymus led to a relative expansion of lung mimetic and Ccl21+ immature mTECs at the expense of MHCII-hi Aire-expressing and all other mimetic populations. Consistent with this, the disruption of mTEC development had functional consequences, including alterations in tuft-associated iNKT polarization and microfold-associated B-cell class switching. In addition, high-resolution transcriptomics revealed that Fezf2 and Aire regulate distinct transcriptional programs, with Fezf2 driving both activation and repression of a more limited set of tissue-restricted genes compared to the broad gene activation program observed for Aire. Pure transcriptional repression at Fezf2 target loci was sufficient to partially rescue mTEC development in global Fezf2-/- (tKO) mice, potentially through suppression of Lifr expression and modulation of downstream Stat3 signaling tone. These results expand our understanding of Fezf2 in mTEC biology, highlighting transcriptional repression as a required functional facet for adult steady-state lineage patterning across the mTEC compartment.
]]></description>
<dc:creator>Germino, J.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Balolong, J.</dc:creator>
<dc:creator>Tang, V.</dc:creator>
<dc:creator>Sun, I.-H.</dc:creator>
<dc:creator>Nie, Y.</dc:creator>
<dc:creator>Tsyporin, J.</dc:creator>
<dc:creator>Proekt, I.</dc:creator>
<dc:creator>Rattay, K.</dc:creator>
<dc:creator>Ntranos, V.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>Chen, B.</dc:creator>
<dc:creator>Gardner, J. M.</dc:creator>
<dc:creator>Miller, C. N.</dc:creator>
<dc:date>2026-01-01</dc:date>
<dc:identifier>doi:10.64898/2025.12.31.697099</dc:identifier>
<dc:title><![CDATA[Fezf2 regulates differentiation of Aire-expressing and post-Aire mimetic epithelial populations maintaining thymic homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.31.697063v1?rss=1">
<title>
<![CDATA[
Progenitor Diversity and Architecture of the Human Ganglionic Eminences Shaping the Basal Ganglia 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.31.697063v1?rss=1</link>
<description><![CDATA[
The embryonic medial and lateral ganglionic eminences (MGE, LGE) are the principal sources of most neurons and glia for the basal ganglia. In primates, the MGE has a distinctive cytoarchitecture characterized by doublecortin enriched cellular nests (DENs), yet the architectonic organization underlying DEN formation, the molecular heterogeneity of ganglionic eminence progenitors and their lineage relationships, remain poorly understood. Here, using paired single-nucleus transcriptomics and chromatin accessibility profiling of the three GEs, we identify distinct progenitor populations, delineate their gene regulatory networks, and reconstruct their lineage trajectories. Live imaging reveals a unipolar outer radial glia-like population (GE-oRG) that undergoes mitotic somal translocation. Spatial transcriptomics identifies a distinct CRABP1+/ANGPT2+ domain within the MGE. Integrated spatial and electron microscopy demonstrates a periphery-to-center gradient of differentiation in the MGE. Leveraging DEN-forming MGE organoids derived from PCDH19 knockout human pluripotent stem cell lines, we identify the protocadherin, PCDH19, as a key regulator of DEN formation.
]]></description>
<dc:creator>Siebert, C. V.</dc:creator>
<dc:creator>Song, M.</dc:creator>
<dc:creator>Moriano, J. A.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Silla, A. C.</dc:creator>
<dc:creator>Walker, M.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Baltazar, J.</dc:creator>
<dc:creator>Oliveira, L. G. d.</dc:creator>
<dc:creator>Shankar, M.</dc:creator>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Suraparaju, P.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Bi, Q.</dc:creator>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Ren, Y.</dc:creator>
<dc:creator>Garcia, M. T.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Zuo, G.</dc:creator>
<dc:creator>Smidt, M. P.</dc:creator>
<dc:creator>Hoekman, M. F. M.</dc:creator>
<dc:creator>Harwell, C.</dc:creator>
<dc:creator>Parent, J.</dc:creator>
<dc:creator>Rubenstein, J.</dc:creator>
<dc:creator>Alvarez-Buylla, A.</dc:creator>
<dc:creator>Kriegstein, A.</dc:creator>
<dc:date>2026-01-02</dc:date>
<dc:identifier>doi:10.64898/2025.12.31.697063</dc:identifier>
<dc:title><![CDATA[Progenitor Diversity and Architecture of the Human Ganglionic Eminences Shaping the Basal Ganglia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.31.697241v1?rss=1">
<title>
<![CDATA[
Single cell RNA-sequencing reveals an association between testosterone treatment and reduced hormone signaling in the human mammary gland 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.31.697241v1?rss=1</link>
<description><![CDATA[
Testosterone is the most studied androgen in hormone replacement therapy in postmenopausal women. Many transgender men also receive long-term testosterone replacement therapy (TRT) and have high serum testosterone levels compared to the low levels seen in cisgender women. Compared to other sex hormones such as estrogen and progesterone, the effects of testosterone on the mammary gland have been relatively understudied and there is little data regarding the long term safety of this treatment. Comparison of mammary glands from transgender men on TRT and cisgender women can reveal the effects of testosterone treatment on mammary gland biology and provide critical information regarding the long-term effects of TRT on patient health and disease outcomes. In this study, we performed single-cell RNA sequencing of breast tissues from a demographics-matched cohort of cisgender women and transgender men on TRT. Surprisingly, participants on TRT had unchanged serum levels of estradiol compared to controls. Among the observed transcriptional differences for participants on TRT were a dramatically reduced expression of genes downstream of estrogen signaling pathways in hormone receptor positive (HR+) luminal epithelial cells, as well as a decreased overall menstrual cycle-related hormone signaling. We confirmed this finding experimentally by showing reduced expression of progesterone receptor a/b, a prominent marker of estrogen signaling, in donors on TRT. Our results support the hypothesis that high levels of testosterone in transgender men on TRT suppress sex hormone signaling in the breast as seen on their impact on HR+ mammary epithelial cells, with implications for TRT as an antagonist of estrogen signaling and protection against breast cancer.
]]></description>
<dc:creator>Phong, K. T.</dc:creator>
<dc:creator>Song, S.</dc:creator>
<dc:creator>Kim, E. A.</dc:creator>
<dc:creator>Conrad, D. N.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:date>2026-01-02</dc:date>
<dc:identifier>doi:10.64898/2025.12.31.697241</dc:identifier>
<dc:title><![CDATA[Single cell RNA-sequencing reveals an association between testosterone treatment and reduced hormone signaling in the human mammary gland]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.01.697272v1?rss=1">
<title>
<![CDATA[
Meta-learning is expressed through altered prefrontal cortical dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.01.697272v1?rss=1</link>
<description><![CDATA[
Learning where and when rewards like food and water are available is essential for survival1,2. In the simplest cases where resource availability is stable, animals can learn reward contingencies by integrating outcomes across repeated samples of each possible action. In more natural settings, however, reward availability is governed by structured higher-order rules such as depletion and repletion over time. To adapt flexibly to such changing environments, optimal choices require meta-learning wherein animals learn how to learn from external feedback, ultimately enabling them to infer the underlying reward structure from abstract, generalizable rules rather than relying solely on recent outcomes3,4. The existence of meta-learning in animal behavior is well established3-8, yet the neural circuits and computations that implement it remain poorly understood9-11. Here we investigated meta-learning using a spatial foraging task in which rats acquired a depletion-repletion rule that regulated reward availability, and carried out longitudinal, high-density recordings from the medial prefrontal cortex (mPFC). We show that meta-learning engages specific, systematic changes in mPFC neural dynamics that embed the learned rule and thereby alter how the network learns action values from reward outcomes. These dynamics are based on mixed coding of task structure and value in individual mPFC neurons. At the population level, this coding organizes into low-dimensional dynamical motifs that generalize across task conditions. As meta-learning progresses, these motifs are reshaped to instantiate both rule-guided inference of future states before outcome delivery and rule-based value updating during the outcome period. These results indicate that meta-learning sculpts pre-existing prefrontal dynamics to support the acquisition of new, generalizable reward-learning strategies.
]]></description>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Comrie, A. E.</dc:creator>
<dc:creator>Kahn, A. E.</dc:creator>
<dc:creator>Monroe, E. J.</dc:creator>
<dc:creator>Joshi, A.</dc:creator>
<dc:creator>Guidera, J. A.</dc:creator>
<dc:creator>Denovellis, E. L.</dc:creator>
<dc:creator>Krausz, T. A.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Thompson, P.</dc:creator>
<dc:creator>Hernandez, J.</dc:creator>
<dc:creator>Yorita, A.</dc:creator>
<dc:creator>Haque, R.</dc:creator>
<dc:creator>Berke, J. D.</dc:creator>
<dc:creator>Daw, N. D.</dc:creator>
<dc:creator>Frank, L. M.</dc:creator>
<dc:date>2026-01-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.01.697272</dc:identifier>
<dc:title><![CDATA[Meta-learning is expressed through altered prefrontal cortical dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.02.697443v1?rss=1">
<title>
<![CDATA[
Conformational changes upon pore blocker removal reveal conductive states of TMEM16A 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.02.697443v1?rss=1</link>
<description><![CDATA[
TMEM16A is a Ca2+-activated anion channel that provides direct electrical feedback to the plasma membrane in response to intracellular Ca2+. Its conductive state remains unresolved, leaving questions about gating, Cl- permeation, and modulation by Ca2+, depolarization, and lipids. To investigate the open state, we performed molecular dynamics simulations of TMEM16A bound to the putative open-state blocker 1PBC. After inhibitor removal, the putative, pore-lining helix TM4 developed kinks at two sites: an upper site that opens the pore for Cl- permeation, and a deeper site causing constriction. A conserved hydrophobic network between TM3 and TM4 persisted in most open structures but separated during extreme dilation, allowing lipids to transiently block the pore. Patch-clamp recordings indicated that the intact network promotes activation. Further simulations yielded >60 Cl- permeation events and a single-channel conductance matching experiments. Additional electrostatic and kinetic modeling indicated that TMEM16As transition from outward-rectification to Ohmic conductance with increasing Ca2+ results from a weak voltage dependence of Ca2+ binding, which act cooperatively to open the pore.

HIGHLIGHTSO_LIRemoval of the inhibitor 1PBC from TMEM16A induces spontaneous transitions to ion conductive and non-conductive states via bending in TM4 at two different locations.
C_LIO_LISimulations suggest that the open state is stabilized by a small hydrophobic network between TM3 and 4 and disrupting this network biases channel closure in electrophysiological recordings.
C_LIO_LIA kinetic model of conduction based on energetics from the all-atom MD simulations coupled to continuum calculations gives a linear current-voltage curve consistent with the fully open conformation and removal of 1 Ca2+ switches to an outwardly rectifying state via electrostatic influence on the Cl- energy profile. However, the rectification is too weak to match experiment, but a cooperative model of Ca2+ binding with weak voltage-dependence does match experiment.
C_LI
]]></description>
<dc:creator>Stephens, C. A.</dc:creator>
<dc:creator>Marcoline, F. V.</dc:creator>
<dc:creator>Peters, C. J.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:date>2026-01-03</dc:date>
<dc:identifier>doi:10.64898/2026.01.02.697443</dc:identifier>
<dc:title><![CDATA[Conformational changes upon pore blocker removal reveal conductive states of TMEM16A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.05.697388v1?rss=1">
<title>
<![CDATA[
Ninhydrin as a covalent warhead for chemical proteomic-enabled discovery and selective engagement of reactive arginines 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.05.697388v1?rss=1</link>
<description><![CDATA[
Covalent molecules have emerged as next-generation therapeutics and as powerful tools for perturbing fundamental biological processes. Chemical proteomic methods to screen for reactive proteinaceous amino acids have transformed small-molecule discovery pipelines, but their application remains mostly limited to sites where reactive cysteines and lysines are present. Here we report a ninhydrin-based warhead that selectively modifies arginine residues, thus expanding the repertoire of amino acids targetable by covalent molecules. Specifically, we developed alkyne-functionalized variants of ninhydrin to establish an arginine-specific chemical proteomics platform, enabling the classification of more than 6,800 unique reactive arginines. These studies uncovered potential modification sites on disease-relevant proteins, including reactive arginines within catalytic sites that are essential for function. By endowing a reversible small molecule inhibitor of cyclophilin A with a ninhydrin warhead, we achieved selective, covalent engagement, and attenuation of enzymatic activity, highlighting the potential for targeting arginines in future therapeutic development campaigns. These findings establish ninhydrin as a warhead for studying arginine reactivity and modulating protein function.
]]></description>
<dc:creator>Ecker, A.</dc:creator>
<dc:creator>Langen, A.</dc:creator>
<dc:creator>Fields, C.</dc:creator>
<dc:creator>Montano, J. L.</dc:creator>
<dc:creator>Tran, M.</dc:creator>
<dc:creator>Seiple, I. B.</dc:creator>
<dc:creator>Zaro, B. W.</dc:creator>
<dc:date>2026-01-05</dc:date>
<dc:identifier>doi:10.64898/2026.01.05.697388</dc:identifier>
<dc:title><![CDATA[Ninhydrin as a covalent warhead for chemical proteomic-enabled discovery and selective engagement of reactive arginines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.05.697753v1?rss=1">
<title>
<![CDATA[
HONeD-in on Brain Activity: Deconvolving Passive Diffusion on the Structural Network from Functional Brain Signals 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.05.697753v1?rss=1</link>
<description><![CDATA[
Brain regions perform distinct computations, and their signals propagate through the whole-brain white matter network. Yet, mathematical models that describe this signal propagation via purely passive diffusion can predict a considerable amount of the observed functional connectivity between regions. This raises a critical question: if so much functional connectivity can be explained by a passive process, how can we isolate the active process? Here, we calculate in closed-form an estimate for such an active signal in functional MRI by spatially deconvolving the effect of passive signal spread over the brains structural connectivity using a higher-order network diffusion (HONeD) model. Across 770 Human Connectome Project subjects, we show that the resulting HONeD-innovation (HONeD-in) signal 1) sparsifies functional connectivity while retaining a well-connected network, 2) remodels resting-state networks (RSNs), 3) mixes the unimodal- multimodal hierarchical organization of RSNs into a circle with no clear hierarchy, and 4) deblurs task-activation maps. Together, our results highlight HONeD deconvolution as a generalizable new way to study resting-state and task fMRI brain signals.
]]></description>
<dc:creator>Sipes, B. S.</dc:creator>
<dc:creator>Arab, F.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:date>2026-01-05</dc:date>
<dc:identifier>doi:10.64898/2026.01.05.697753</dc:identifier>
<dc:title><![CDATA[HONeD-in on Brain Activity: Deconvolving Passive Diffusion on the Structural Network from Functional Brain Signals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.05.696859v1?rss=1">
<title>
<![CDATA[
Cellular consequences, citrullination substrates, and antigenicity resulting from wild-type and targeted PAD4 on cell surfaces 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.05.696859v1?rss=1</link>
<description><![CDATA[
Protein arginine deiminase-4 (PAD4) catalyzes hydrolysis of arginine to citrulline in proteins that promotes widespread changes in cellular phenotypes through transcriptional regulation that can induce innate immunity and promote cancer. Overexpression and hyperactivity of PAD4 leads to a form of cell death called NETosis that releases PAD4 to the extracellular space. In excess, release of PAD4 is believed to be a major cause of various autoimmune diseases through the generation of anti-citrulline protein antibodies (ACPAs). Little is known about the specific protein substrates that become citrullinated and lead to autoimmunity, but there is growing evidence that PAD4 can be localized to the cell surface in response to inflammation. Here, we further characterize the cellular consequences for exogenous treatment with PAD4 showing that it induces morphological changes that increase cell migration, a hallmark of cancer. We then devised a more simplified and robust proteomics approach to identify PAD4 substrates. We identified some 1000 endogenously citrullinated peptides from 500 proteins, and 3000 citrullinated peptides from 1300 proteins upon exogenous addition of PAD4 both inside and outside of cells. This extracellular set can be further augmented by targeting PAD4 to a cancer target, HER2, using a binding protein conjugate. Finally, we studied how citrullinated cells can induce a robust humoral response in a syngeneic vaccine model to produce ACPAs. We believe these studies further our understanding of cell phenotypic consequences of extracellular PAD4 and new PAD4 substrates both inside and outside of cells that are potential neoepitopes for generation of ACPAs.
]]></description>
<dc:creator>Kong, S.</dc:creator>
<dc:creator>Peters-Clarke, T. M.</dc:creator>
<dc:creator>Delaveris, C. S.</dc:creator>
<dc:creator>Phojanakong, P.</dc:creator>
<dc:creator>Steri, V.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2026-01-06</dc:date>
<dc:identifier>doi:10.64898/2026.01.05.696859</dc:identifier>
<dc:title><![CDATA[Cellular consequences, citrullination substrates, and antigenicity resulting from wild-type and targeted PAD4 on cell surfaces]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.07.698158v1?rss=1">
<title>
<![CDATA[
Protein tyrosine phosphatase receptor type kappa (PTPRκ) regulates Superior ON-Direction Selective Ganglion Cell development, facilitating image stabilization 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.07.698158v1?rss=1</link>
<description><![CDATA[
In vertebrates, gaze stabilization during global visual motion requires ON direction-selective ganglion cells (oDSGCs) in the Accessory Optic System (AOS) to drive the optokinetic reflex (OKR). Three types of oDSGCs form independent mosaics; however, the mechanisms that specify and maintain their numbers to create these mosaics during development remain unknown. Here, we identify protein tyrosine phosphatase receptor type kappa (Ptprk) as a key regulator of Superior oDSGC density, the subtype tuned to upward motion. High-depth single-cell RNA sequencing (scRNAseq) reveals that Ptprk is selectively enriched in Superior oDSGCs compared to Inferior oDSGCs, which detect downward motion. Genetic deletion of Ptprk selectively halves the number of Superior oDSGCs while preserving the number of Inferior oDSGCs. Anatomically, Superior oDSGCs in Ptprk mutants exhibit compensatory neurite remodeling to preserve mosaics. Physiologically, oDSGCs have broader tuning curves, and a subset of Superior oDSGCS have aberrant preferred directions in Ptprk mutants. Postsynaptically, loss of Ptprk reduces oDSGC connectivity to central targets. Behaviorally, the OKR responses to upward motion are attenuated in both global and pan-retinal Ptprk mutants, confirming a retinal role for Ptprk in upward image stabilization. Together, these findings demonstrate that the density of a retinal ganglion cell type is under molecular control, and that reducing density drives neurite remodeling, alters circuit computation, and ultimately impairs visually-guided behavior.
]]></description>
<dc:creator>Lin, T.-H.</dc:creator>
<dc:creator>Balraj, A.</dc:creator>
<dc:creator>Al-Khindi, T.</dc:creator>
<dc:creator>Kiraly, J. K.</dc:creator>
<dc:creator>Dunn, F. A.</dc:creator>
<dc:creator>Kolodkin, A. L.</dc:creator>
<dc:date>2026-01-07</dc:date>
<dc:identifier>doi:10.64898/2026.01.07.698158</dc:identifier>
<dc:title><![CDATA[Protein tyrosine phosphatase receptor type kappa (PTPRκ) regulates Superior ON-Direction Selective Ganglion Cell development, facilitating image stabilization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.07.698214v1?rss=1">
<title>
<![CDATA[
A Joint Tau Propagation and Neuroinflammation Model Reinforces Inflammatory Modulation of Network-Driven Spread in Alzheimer's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.07.698214v1?rss=1</link>
<description><![CDATA[
Alzheimers disease (AD) is a progressive neurodegenerative disorder and the leading cause of dementia. Despite over a century of research, AD remains untreatable due to an incomplete understanding of its underlying mechanisms. While amyloid beta has dominated therapeutic efforts, tau pathology and neuroinflammation represent critical disease drivers and intriguing therapeutic targets. We developed an extended computational model building upon the open-source Aggregation Network Diffusion (AND) framework by coupling spatial tau propagation and aggregation with ordinary differential equations representing key inflammatory cascades. The model incorporates M1/M2 microglia and astrocytic activation, cytokine-mediated feedback loops, and neuronal loss, all modulated via region-specific genetic expression matrices of ApoE and TREM2 with variant-specific weighting. The model maintains biologically plausible tau dynamics while generating robust inflammatory marker trends, serving as a computational testbed for hypothesis generation and mechanistic exploration. The inflammatory components independently capture experimental observations and display a marked M1/M2 microglia divergence. The model demonstrates enhanced similarity to regional tau propagation patterns due to inflammatory and genetic components, reinforcing neuroinflammations role in tau spread and highlighting the need to incorporate these processes in modeling efforts. Finally, through parsimony analysis, we identify microglia and pro-inflammatory rates (including microglia-facilitated tau spread) as key contributors to improved model accuracy, informing future modeling approaches.
]]></description>
<dc:creator>Dhaliwal, I.</dc:creator>
<dc:creator>Sandell, R.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:date>2026-01-08</dc:date>
<dc:identifier>doi:10.64898/2026.01.07.698214</dc:identifier>
<dc:title><![CDATA[A Joint Tau Propagation and Neuroinflammation Model Reinforces Inflammatory Modulation of Network-Driven Spread in Alzheimer's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.08.698381v1?rss=1">
<title>
<![CDATA[
Targeted Chromosomal Sequencing of Wild Bonobos Identifies a Genetically Distinct Subpopulation East of the Lomami River 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.08.698381v1?rss=1</link>
<description><![CDATA[
Bonobos (Pan paniscus), an endangered species, have for decades been genetically understudied, partly due to difficulties in obtaining high-quality samples. The study of their genome is important not only for understanding their evolution, but also for improving conservation efforts, including population management, diversity and inbreeding assessment, and tracking rescued individuals to combat illegal wildlife trafficking.

Here, we use chromosome 21 target capture data from 156 non-invasively collected faecal samples from wild bonobos to perform a comprehensive analysis of their population structure. We confirm the existence of three previously suggested subpopulations identified here as Western, Central and Eastern bonobos which are defined by natural barriers of gene flow such as the Lomami River. By estimating levels of inbreeding, diversity and differentiation, we find support for isolation of mainly Western and Eastern populations and add information on the dispersal routes of their ancestors. We infer split times and separation of these populations and apply a genetic framework to geolocalize samples of unknown origin, showing that locations of their potential origin can be estimated with a precision of up to a median of [~]50 km. Our study provides valuable insight into the evolution and population structure of bonobos and reveals how rivers act as strong barriers between populations. It also offers resources for conservation efforts and highlights the need to monitor bonobo populations more closely, in particular isolated ones.

ImpactBonobos have been difficult to study genetically due to their remote forest habitat and endangered status. Here we used non-invasive sampling and chromosome 21 target capture sequencing to perform the most detailed analysis to date of their population structure. We find three main subpopulations and genetic differentiation influenced by river barriers, especially with populations found on the Eastern side of the Lomami river. This data will be useful for identifying the geographic origin of confiscated samples and thus aid bonobo conservation efforts.
]]></description>
<dc:creator>Crego-Walters, M.</dc:creator>
<dc:creator>Cuadros-Espinoza, S.</dc:creator>
<dc:creator>Ruiz-Gartzia, I.</dc:creator>
<dc:creator>Han, S.</dc:creator>
<dc:creator>Hermosilla, N.</dc:creator>
<dc:creator>Helsen, P.</dc:creator>
<dc:creator>Frandsen, P.</dc:creator>
<dc:creator>Brumwell Prats, A. M.</dc:creator>
<dc:creator>Alentorn-Moron, P.</dc:creator>
<dc:creator>Fontsere, C.</dc:creator>
<dc:creator>Alvarez Estape, M.</dc:creator>
<dc:creator>Ngofuna, M.</dc:creator>
<dc:creator>Monghiemo, C.</dc:creator>
<dc:creator>Leendertz, F.</dc:creator>
<dc:creator>Thompson, J.</dc:creator>
<dc:creator>Fasbender, D.</dc:creator>
<dc:creator>Dieguez, P.</dc:creator>
<dc:creator>Lotana Lokasola, A.</dc:creator>
<dc:creator>Brand, C.</dc:creator>
<dc:creator>Ndjango, J.-B.</dc:creator>
<dc:creator>Georgiev, A. V.</dc:creator>
<dc:creator>Giles, J. A.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Sharp, P. M.</dc:creator>
<dc:creator>Pereboom, Z.</dc:creator>
<dc:creator>Andres, A. M.</dc:creator>
<dc:creator>Kuhlwilm, M.</dc:creator>
<dc:creator>Gronau, I.</dc:creator>
<dc:creator>Kuehl, H.</dc:creator>
<dc:creator>Wessling, E. G.</dc:creator>
<dc:creator>Narat, V.</dc:creator>
<dc:creator>Surbeck, M.</dc:creator>
<dc:creator>Hart, J. A.</dc:creator>
<dc:creator>Hart, T. B.</dc:creator>
<dc:creator>Hvilsom, C.</dc:creator>
<dc:creator>Krutzen, M.</dc:creator>
<dc:creator>Stevens, J.</dc:creator>
<dc:creator>Hahn, B. H.</dc:creator>
<dc:creator>Lizano, E.</dc:creator>
<dc:creator>Prado-Martinez, J.</dc:creator>
<dc:creator>M</dc:creator>
<dc:date>2026-01-09</dc:date>
<dc:identifier>doi:10.64898/2026.01.08.698381</dc:identifier>
<dc:title><![CDATA[Targeted Chromosomal Sequencing of Wild Bonobos Identifies a Genetically Distinct Subpopulation East of the Lomami River]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.08.698469v1?rss=1">
<title>
<![CDATA[
Smoothened turnover regulated by Hedgehog signaling in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.08.698469v1?rss=1</link>
<description><![CDATA[
During development, the processes of cell-cell signaling orchestrate growth and morphogenesis using both static features and moving parts. For example, expression domains of genes which set states of determination may be fixed in space for periods of time, but multiple signals move between the determined cells and intracellular movements of components of the signal transduction machines are key to information exchange. In the Drosophila wing imaginal disc, Hedgehog (Hh) protein produced in the posterior compartment moves to cells across the anterior/posterior compartment border to induce signal transduction and various intracellular responses. In this work, we investigated protein turnover and intracellular trafficking of components of Hh signal transduction in the wing disc and found that both are major features of Hh signaling.

SummaryThe intracellular distribution and half-life of Smoothened is regulated by Hedgehog signaling.
]]></description>
<dc:creator>Hatori, R.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Kornberg, T.</dc:creator>
<dc:date>2026-01-09</dc:date>
<dc:identifier>doi:10.64898/2026.01.08.698469</dc:identifier>
<dc:title><![CDATA[Smoothened turnover regulated by Hedgehog signaling in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.09.698662v1?rss=1">
<title>
<![CDATA[
Tumor cell death by ferroptosis contributes to an immunosuppressive tumor microenvironment in syngeneic murine models of cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.09.698662v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDAC) is characterized by profound metabolic rewiring and a strongly immunosuppressive tumor microenvironment, both of which contribute to poor therapeutic responses. Immunogenic cell death (ICD) represents a potential strategy to overcome immune suppression by coupling tumor cell death to anti-tumor immune activation. Here, we investigated whether targeting amino acid metabolism in PDAC can induce ICD and promote tumor immunity. Through a focused metabolic screen in a panel of syngeneic mouse cancer cell lines, we identified cysteine restriction as a robust inducer of multiple damage-associated molecular patterns (DAMPs) in vitro, hallmark features of ICD. In addition to driving DAMPs, cystine-deprived tumor cells also promoted dendritic cell phagocytosis, maturation, and proinflammatory cytokine production in vitro. Because cysteine deprivation is a known trigger of ferroptosis, we further demonstrated that pharmacologic inhibition of glutathione peroxidase 4 (GPX4) similarly elicited ICD-associated features, which were reversible by the ferroptosis inhibitor Ferrostatin-1.

To define additional immune-modulatory signals associated with ferroptosis, we performed metabolomic and lipidomic profiling of cells undergoing, but not yet committed to, ferroptotic death. These analyses revealed selective release of immunosuppressive metabolites and oxidized phospholipids. Consistent with this, conditioned media from ferroptotic cells impaired CD8 T cell proliferation and cytotoxicity in vitro. Thus, together our results indicated that the induction of ferroptotic immunogenic cell death led to the release of both pro- and anti-inflammatory signals. Subsequent analysis in vivo revealed that ferroptotic tumor cells predominantly contributed to a tumor-protective environment. In particular, tumors inoculated with ferroptotic cells were enriched with immunosuppressive myeloid cells and exhibited reduced populations of tumor-infiltrating CD8+ T cells. Further investigation using immune compromised mice suggested that ferroptotic cells may suppress both adaptive and innate immune responses. Collectively, these results underscore the complex and highly context-dependent effects of ferroptosis on tumor immunity, highlighting the critical importance of in vivo models to determine true immunogenic potential within the tumor microenvironment.
]]></description>
<dc:creator>Mbah, N. E.</dc:creator>
<dc:creator>Sutton, D.</dc:creator>
<dc:creator>Hong, H. S.</dc:creator>
<dc:creator>Singhal, R.</dc:creator>
<dc:creator>Menjivar, R. E.</dc:creator>
<dc:creator>Giza, H.</dc:creator>
<dc:creator>Sajjakulnukit, P.</dc:creator>
<dc:creator>Perricone, M.</dc:creator>
<dc:creator>Nwosu, Z.</dc:creator>
<dc:creator>Alektiar, J.</dc:creator>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Long, D.</dc:creator>
<dc:creator>Andren, A. C.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Crawford, H. C.</dc:creator>
<dc:creator>Frankel, T. L.</dc:creator>
<dc:creator>Pasca di Magliano, M.</dc:creator>
<dc:creator>Franchi, L.</dc:creator>
<dc:creator>Shah, Y. M.</dc:creator>
<dc:creator>Lyssiotis, C. A.</dc:creator>
<dc:date>2026-01-11</dc:date>
<dc:identifier>doi:10.64898/2026.01.09.698662</dc:identifier>
<dc:title><![CDATA[Tumor cell death by ferroptosis contributes to an immunosuppressive tumor microenvironment in syngeneic murine models of cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.10.698836v1?rss=1">
<title>
<![CDATA[
Atypical GPCR Activation Resolved by Nanobody Engineering 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.10.698836v1?rss=1</link>
<description><![CDATA[
Abstract/ Summary ParagraphG protein-coupled receptors (GPCRs) are the largest family of clinically targeted proteins, yet most therapeutics target a narrow subset of structurally well-behaved receptors. The atypical chemokine receptor ACKR3 defies canonical models, displaying broad ligand recognition, high basal activity, and resistance to inhibition. Using engineered nanobodies, cryo-EM, NMR, and structure-guided pharmacology, we uncover an unconventional activation mechanism in ACKR3 that challenges established paradigms of GPCR activation. We find that receptor activity is controlled by changes in extracellular pocket volume rather than conformational rearrangements in conserved microswitches, and an expanded aromatic cluster at the intracellular transducer binding pocket stabilizes the active state. These findings redefine how GPCRs can be modulated and open new strategies for targeting pharmacologically intractable receptors.
]]></description>
<dc:creator>Schlimgen, R. R.</dc:creator>
<dc:creator>Jenjak, S. E.</dc:creator>
<dc:creator>De La Sancha, A.</dc:creator>
<dc:creator>Darcis, J.</dc:creator>
<dc:creator>Billesboelle, C. B.</dc:creator>
<dc:creator>Olson, L. J.</dc:creator>
<dc:creator>Peterson, F. C.</dc:creator>
<dc:creator>Smit, M. J.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:creator>Szpakowska, M.</dc:creator>
<dc:creator>Chevigne, A.</dc:creator>
<dc:creator>Volkman, B. F.</dc:creator>
<dc:date>2026-01-11</dc:date>
<dc:identifier>doi:10.64898/2026.01.10.698836</dc:identifier>
<dc:title><![CDATA[Atypical GPCR Activation Resolved by Nanobody Engineering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.10.698783v1?rss=1">
<title>
<![CDATA[
Intramolecular loops control SARS-CoV-2 nucleocapsid protein self-association and nucleic acid binding dependent on phosphorylation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.10.698783v1?rss=1</link>
<description><![CDATA[
The nucleocapsid protein of SARS-CoV-2 scaffolds genomic RNA into ribonucleoprotein complexes (RNP) for assembly in the virion, but also fulfills critical intracellular functions in replication and the suppression of host defense. It contains a folded nucleic acid binding domain (NTD) and a dimerization domain, connected by a disordered linker containing a serine/arginine-rich (SR) region and a leucine-rich sequence (LRS). The switch between intracellular and assembly functions is controlled by phosphorylation of the SR region, but the molecular details are unclear. Here we describe a model in which two mutually exclusive intramolecular loops bind the NTD and dynamically control self-association and nucleic acid binding properties dependent on the SR linker phosphorylation state. The model is supported by biophysical measurements of hydrodynamic radii, reversible protein self-association, nucleic acid binding, and thermodynamic stability, as well as structural predictions and published NMR chemical shifts. We find SR linker phosphorylation compacts the protein and inhibits nucleic acid binding and RNP formation, while enhancing self-association through promotion of transient coiled-coils in the LRS of the linker. These changes shift the nucleocapsid protein to a configuration poised for multi-valent interactions that support intracellular functions.
]]></description>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Datta, S. A.</dc:creator>
<dc:creator>Trent, C.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Saleem, Z.</dc:creator>
<dc:creator>Szczesna, E.</dc:creator>
<dc:creator>Owoborode, F.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Adly, A. N.</dc:creator>
<dc:creator>Kalish, H. R.</dc:creator>
<dc:creator>Piszczek, G.</dc:creator>
<dc:creator>Morgan, D. O.</dc:creator>
<dc:creator>Schuck, P.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:date>2026-01-12</dc:date>
<dc:identifier>doi:10.64898/2026.01.10.698783</dc:identifier>
<dc:title><![CDATA[Intramolecular loops control SARS-CoV-2 nucleocapsid protein self-association and nucleic acid binding dependent on phosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.11.697334v1?rss=1">
<title>
<![CDATA[
PRSS56 acts as an intrinsic retinal signal driving postnatal ocular axial growth and myopia susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.11.697334v1?rss=1</link>
<description><![CDATA[
Myopia is a leading cause of visual impairment worldwide, and high myopia markedly increases the risk of irreversible vision loss. Although visual experience guides postnatal ocular elongation, the role of intrinsic retinal growth signals remains poorly defined. Here we identify the serine protease PRSS56 as a retinal factor that promotes ocular axial growth beyond early development. Using genetic mouse models, we show that conditional inactivation of Prss56 in Muller glia reduces axial length and causes hyperopia even under dark-rearing conditions, demonstrating that PRSS56 drives axial elongation independently of light-evoked visual input during emmetropization. Conversely, Muller glia-specific overexpression of Prss56 induces axial elongation in a proteolysis-dependent manner, supporting its role as an autonomous retinal growth signal. In concordance, human genetic analyses reveal that the common PRSS56 variant rs2853447 is associated with increased axial length in individuals with myopia and high myopia, but not in non-myopes, suggesting that this variant confers a selective growth advantage in individuals predisposed to ocular elongation. Functional genomic analyses further identify a myopia-associated variant, rs2741297, within a retinal enhancer in intron 4 of PRSS56 marked by open chromatin and transcription factor occupancy. Together, these findings establish PRSS56 as an intrinsic retinal growth factor that functions beyond early eye development and support a model in which genetic and environmental factors converge on retinal pathways to modulate myopia susceptibility.
]]></description>
<dc:creator>Gangappa, K.</dc:creator>
<dc:creator>Duraikannu, D.</dc:creator>
<dc:creator>Advani, J.</dc:creator>
<dc:creator>Labelle-Dumais, C.</dc:creator>
<dc:creator>Uludag, E. B.</dc:creator>
<dc:creator>Koli, S.</dc:creator>
<dc:creator>Singh, N.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Kuo, Y.-M.</dc:creator>
<dc:creator>Ishikawa, Y.</dc:creator>
<dc:creator>Choquet, H.</dc:creator>
<dc:creator>Swaroop, A.</dc:creator>
<dc:creator>Nair, K. S.</dc:creator>
<dc:date>2026-01-12</dc:date>
<dc:identifier>doi:10.64898/2026.01.11.697334</dc:identifier>
<dc:title><![CDATA[PRSS56 acts as an intrinsic retinal signal driving postnatal ocular axial growth and myopia susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.12.698936v1?rss=1">
<title>
<![CDATA[
Osteoprotegerin-Enabled Immune Evasion by Pathological Adipose Stromal Cells Drives Metabolic Dysfunction in Obesity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.12.698936v1?rss=1</link>
<description><![CDATA[
Diet-induced obesity (DIO) promotes the accumulation of stromal cells with senescent characteristics in the adipose tissue (AT). Selectively clearing these cells--either through chemical senolytics or activation of invariant natural killer T (iNKT) cells--improves glucose homeostasis in obese mice, however the identity of the responsible stromal population remains unknown. Here, we use transcriptional profiling of AT stromal cells coupled with C12FDG-based enrichment of senescent populations in mice with DIO and healthy controls to identify a distinct subset of adipose progenitor cells (APCs) with robust senescence signatures that accumulate in DIO across multiple AT depots. We show that these cells, which we term senescent APCs (sAPCs), are not merely passive markers of metabolic stress but are instead active stromal organizers, accumulating in parallel with the emergence of lipid-associated macrophages (LAMs) and the diminution of multipotent mesenchymal progenitors. sAPCs promoted CCR2-dependent macrophage chemotaxis, directly linking stromal senescence to chemokine-mediated remodeling of the AT immune niche. Comparative transcriptional analysis revealed a remarkable similarity between sAPCs and inflammatory cancer-associated fibroblasts (iCAFs), including the strong induction of periostin (POSTN) and the production of osteoprotegerin (OPG), a decoy receptor for RANKL and TRAIL that enables tumoral immune evasion. Indeed, OPG production by AT stromal cells was induced by DIO across AT depots. Exogenous OPG inhibited the ability of iNKT cells to kill senescent APCs in vitro, whereas antibody-mediated OPG neutralization reciprocally enhanced such cytotoxic killing. In vivo, systemic OPG neutralization both reduced sAPC accumulation in AT and normalized glucose homeostasis in obese mice. Together, these findings identify sAPCs as a pathological stromal population that expands in obesity through elaboration of immunomodulatory factors. In particular, secreted OPG enables sAPCs to evade iNKT-mediated immune surveillance and contributes to metabolic dysfunction, highlighting OPG and sAPCs as promising therapeutic targets for restoring AT immune and metabolic homeostasis.
]]></description>
<dc:creator>Apolostolopoulou, H.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Bolus, W. R.</dc:creator>
<dc:creator>Singh, R. D.</dc:creator>
<dc:creator>Bains, A.</dc:creator>
<dc:creator>Bahadure, P.</dc:creator>
<dc:creator>Gupta, K.</dc:creator>
<dc:creator>Koliwad, S. K.</dc:creator>
<dc:creator>Bhushan, A.</dc:creator>
<dc:date>2026-01-13</dc:date>
<dc:identifier>doi:10.64898/2026.01.12.698936</dc:identifier>
<dc:title><![CDATA[Osteoprotegerin-Enabled Immune Evasion by Pathological Adipose Stromal Cells Drives Metabolic Dysfunction in Obesity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.12.698943v1?rss=1">
<title>
<![CDATA[
An engineered nanobody inhibitor for molecular-to-circuit control of opioid receptor function 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.12.698943v1?rss=1</link>
<description><![CDATA[
Opioid receptors (ORs) orchestrate pain relief, reward, and dependence, yet their signaling arises from diverse cell types and subcellular compartments that cannot be selectively interrogated with existing pharmacological or genetic approaches. Single-domain antibodies, or nanobodies (Nbs), can probe receptor states, but their potential as tools for controlling native receptor signaling at the system level has remained unexplored. Here, we engineer a suite of high-affinity intracellular Nbs that bind active ORs through structure-guided evolution and in silico design. Iterative optimization yields Nb64, a potent inhibitor that rapidly suppresses transducer engagement, receptor internalization, and downstream signaling, including endogenous pathways in neuronal cells. Organelle targeting highlights Nb64s capacity to control OR activity with subcellular precision, while bio-reversible cell-penetrating peptide (CPP) conjugation enables non-genetic cytosolic delivery. Cell-type-specific expression of Nb64 in VTA interneurons attenuates fentanyl-evoked dopamine release and behavioral responses in mice, demonstrating targeted control of opioid actions in vivo. Nb64 provides a versatile strategy for dissecting OR biology and establishes a generalizable framework for precision inhibition of native GPCR signaling in vivo.
]]></description>
<dc:creator>Valbret, Z.</dc:creator>
<dc:creator>Lamothe-Molina, P. L.</dc:creator>
<dc:creator>Papadogkonaki, S.</dc:creator>
<dc:creator>Koehl, A.</dc:creator>
<dc:creator>Arafiles, J. V. V.</dc:creator>
<dc:creator>Shapiro Tuchman, Z.</dc:creator>
<dc:creator>Fisher, N. M.</dc:creator>
<dc:creator>Radoux-Mergault, A.</dc:creator>
<dc:creator>Canziani, A.</dc:creator>
<dc:creator>Huang, G.</dc:creator>
<dc:creator>Von Zastrow, M.</dc:creator>
<dc:creator>Kosloff, M.</dc:creator>
<dc:creator>Hackenberger, C. P.</dc:creator>
<dc:creator>Manglik, A.</dc:creator>
<dc:creator>Patriarchi, T.</dc:creator>
<dc:creator>Stoeber, M.</dc:creator>
<dc:date>2026-01-14</dc:date>
<dc:identifier>doi:10.64898/2026.01.12.698943</dc:identifier>
<dc:title><![CDATA[An engineered nanobody inhibitor for molecular-to-circuit control of opioid receptor function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.14.699559v1?rss=1">
<title>
<![CDATA[
NPEPPS segmental duplication drives position effect expression of TBC1D3 in the human brain 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.14.699559v1?rss=1</link>
<description><![CDATA[
In humans, the TBC1D3 gene family is thought to play a critical role in the expansion of the frontal cortex by promoting neuronal proliferation during brain development. This gene family shows some of the greatest structural heterozygosity ([~]97%) with haplotype copy numbers ranging from 3-39 among different human haplotypes. This raises the question as to how a gene so crucial in the evolutionary expansion of the human frontal cortex can be so variable in the human population. Here, we characterize the regulatory architecture that explains this paradox. We show that 45-96% of TBC1D3 expression is attributable to a single paralog located at the most telomeric position at the edge of a cluster of TBC1D3 genes. We find that its >3-fold higher expression relative to other copies is driven by a 110 kbp segmental duplication that occurred [~]8.9 million years ago, relocating a partial duplication of the puromycin-sensitive aminopeptidase gene (NPEPPS), including its promoter, adjacent to this TBC1D3 locus. Using neurospheres and comparative transcriptomics of iPSC-derived cultures, we show that expression of NPEPPSP1-TBC1D3 increases as neurons differentiate as a result of alternative splicing and differential polyadenylation usage. While the fusion exists in other ape lineages, we show subsequent deletion of the NPEPPSP1 promoter in Gorilla and a separate, lineage-specific duplication in the Pan lineage ablated the production of this fusion product, rendering this position effect of TBC1D3 specific to humans.
]]></description>
<dc:creator>Guitart, X.</dc:creator>
<dc:creator>Brunner, J. W.</dc:creator>
<dc:creator>Ren, L.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Porubsky, D.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Hoekzema, K.</dc:creator>
<dc:creator>Munson, K. M.</dc:creator>
<dc:creator>Sun, K.</dc:creator>
<dc:creator>Ayllon, M.</dc:creator>
<dc:creator>Hoglin, K.</dc:creator>
<dc:creator>McMullen, R.</dc:creator>
<dc:creator>Pavlovic, B.</dc:creator>
<dc:creator>Pollen, A. A.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:date>2026-01-15</dc:date>
<dc:identifier>doi:10.64898/2026.01.14.699559</dc:identifier>
<dc:title><![CDATA[NPEPPS segmental duplication drives position effect expression of TBC1D3 in the human brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.14.699608v1?rss=1">
<title>
<![CDATA[
Soluble DLK1 secreted by telomere-shortening-induced senescent microglia impairs oligodendrocyte functions and alters neuronal activity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.14.699608v1?rss=1</link>
<description><![CDATA[
Aging is the major risk factor for neurodegenerative disease, yet the mechanisms linking physiological aging to brain dysfunction remain unclear. Because telomere erosion is a hallmark of aging, we examined its impact on glial and neuronal physiology. Telomere shortened mice showed lipofuscinosis, hypomyelination, microglial atrophy, and cognitive deficits. Single nuclei RNA-seq revealed accelerated glial aging, elevated microglial senescence pathways, and impaired oligodendrocyte functions. Inducing senescence in human iPSC derived microglia with shortened telomeres identified soluble DLK1 as a novel senescence associated ligand. sDLK1 was increased in the cerebrospinal fluid of telomere shortened and naturally aged mice, and this increase was eliminated by microglial depletion. AAV delivery of sDLK1 in vivo caused hypomyelination and blocked oligodendrocyte lineage progression, demonstrating the detrimental nature of excessive sDLK1. In human iPSC systems, sDLK1 impaired oligodendrocyte maturation and altered calcium signaling in excitatory neurons. These findings identify microglial senescence as a core consequence of telomere shortening and reveal sDLK1 as a microglia-derived senescence ligand that drives oligodendrocyte and neuronal dysfunction in aging.
]]></description>
<dc:creator>Liu, B.</dc:creator>
<dc:creator>Mahoney, M.</dc:creator>
<dc:creator>Feng, Y.</dc:creator>
<dc:creator>Telpoukhovskaia, M. A.</dc:creator>
<dc:creator>Giani, A.</dc:creator>
<dc:creator>Torres, E. R.</dc:creator>
<dc:creator>Zhan, L.</dc:creator>
<dc:creator>Ye, P.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Foxe, N.</dc:creator>
<dc:creator>Zhu, D.</dc:creator>
<dc:creator>Tong, X.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Theodoris, C. V.</dc:creator>
<dc:creator>Gong, S.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Fan, L.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:date>2026-01-15</dc:date>
<dc:identifier>doi:10.64898/2026.01.14.699608</dc:identifier>
<dc:title><![CDATA[Soluble DLK1 secreted by telomere-shortening-induced senescent microglia impairs oligodendrocyte functions and alters neuronal activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.15.699746v1?rss=1">
<title>
<![CDATA[
B7-H3-targeted natural killer cells effectively kill atypical teratoid / rhabdoid tumors and extend survival in orthotopic xenografts 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.15.699746v1?rss=1</link>
<description><![CDATA[
BackgroundAtypical teratoid/rhabdoid tumors (AT/RTs) are the most common malignant CNS tumor in infants, and patients suffer from low survival rates and treatment-related morbidities. These tumors frequently overexpress the pan-cancer antigen B7-H3 (CD276), which can be targeted with immunotherapy. We hypothesized that adding a B7-H3-targeting cytotoxic chimeric antigen receptor (CAR) to NK cells can enhance killing against AT/RTs.

MethodsWe designed a library of variable affinity B7-H3-targeted CARs, which were transduced into primary healthy donor-derived NK cells. We verified B7-H3 expression in a panel of AT/RT cell lines and further engineered luciferase and nuclear GFP-expressing AT/RT (CHLA-04, CHLA-06, BT12, BT37) as well as a CHLA-06-derived B7-H3 knockout. We tested CAR-NK cell functionality using in vitro co-culture cytotoxicity assays. We delivered anti-B7-H3 CAR-NK cells intratumorally or intracerebroventricularly (ICV) to AT/RT orthotopic xenografts and monitored for tumor growth and animal survival.

ResultsB7-H3-targeted CAR-NK cells demonstrated target-specific cytotoxicity when compared to unmodified NK cells. Knockout of B7-H3 in target cells abolished the increased CAR-mediated target killing. When delivered intratumorally to CHLA-06 orthotopic xenograft-bearing mice, anti-B7-H3 CAR-NK cells eliminated tumor cells and prolonged survival. When CAR-NK cells were delivered ICV against a CNS disseminated tumor model of BT12, treated mice had significantly improved survival.

ConclusionsAnti-B7-H3 CAR-NK cells effectively kill AT/RTs in multiple pre-clinical in vitro and in vivo models in an antigen-specific manner. Evidence of efficacy in translationally relevant models provides support for using B7-H3-targeting CAR-NK cells in high-risk AT/RT patients.
]]></description>
<dc:creator>Choe, T.</dc:creator>
<dc:creator>Chakravarti, S.</dc:creator>
<dc:creator>Holl, N. J.</dc:creator>
<dc:creator>Zinsky, M. T.</dc:creator>
<dc:creator>Jones, D. G.</dc:creator>
<dc:creator>Guchhait, S.</dc:creator>
<dc:creator>Rahnama, R.</dc:creator>
<dc:creator>Vorri, S. C.</dc:creator>
<dc:creator>Amaral, A.</dc:creator>
<dc:creator>Lucas, C.-H. G.</dc:creator>
<dc:creator>Raabe, E. H.</dc:creator>
<dc:creator>Bonifant, C.</dc:creator>
<dc:date>2026-01-16</dc:date>
<dc:identifier>doi:10.64898/2026.01.15.699746</dc:identifier>
<dc:title><![CDATA[B7-H3-targeted natural killer cells effectively kill atypical teratoid / rhabdoid tumors and extend survival in orthotopic xenografts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.16.699530v1?rss=1">
<title>
<![CDATA[
In vivo Proximity & Spatial Proteomics with CRISPR Screening Identify STXBP1 as a Protective Modifier of a-synuclein Toxicity in Dopamine Neurons. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.16.699530v1?rss=1</link>
<description><![CDATA[
Parkinsons disease (PD) is a disease of adults involving the loss of dopaminergic neurons after a long, asymptomatic, prodromal period. -synuclein, LRRK2, and VPS35 are linked to familial PD, however, how these mutations predispose dopamine neurons to death during the early prodromal phases remains unclear. Here, we used in vivo native proximity proteomics (iBioID) and dopaminergic neuron-specific subcellular proteomics across multiple PD models to uncover early alterations preceding neuronal loss. Our analyses identified convergent disruptions in synaptic protein abundance, indicating that presynaptic trafficking defects are early events in PD pathogenesis. Using a targeted CRISPR-based genetic screen in dopamine neurons, we demonstrated that mimicking this misregulation of STXBP1 amplifies vulnerability to -synuclein, implicating it as a previously underappreciated toxicity buffering factor. These findings highlight convergent mechanisms that sensitize dopamine neuronal degeneration and that presynaptic vesicle SNARE-complex proteins could serve as key targets for disease-modifying therapies in PD and related neurodegenerative disorders.

HighlightsO_LIIn vivo native-BioID mapping of multiple Parkinsons disease (PD) protein interactomes revealed a convergent presynaptic network.
C_LIO_LIiBioID analysis on mutant PD proteins (-synuclein A30P, LRRK2 G2019S, VPS35 D620N) uncovered mutation-specific shifts in local proximity networks, notably in endocytic and vesicle recycling pathways.
C_LIO_LISpatial proteomics (iBioCoFrac) of dopamine neurons in vivo identified functional modules with reduced levels of key synaptic proteins in PD models.
C_LIO_LIComparative proteomics using iBioCoFrac revealed synaptic vesicle regulation as a primary site of molecular convergence and early molecular signatures in dopamine neurons across multiple PD mouse models.
C_LIO_LIAn in vivo CRISPR screen pinpointed the presynaptic protein Stxbp1/Munc18-1 as an -synuclein toxicity modifier in dopaminergic neurons.
C_LI
]]></description>
<dc:creator>Shonai, D.</dc:creator>
<dc:creator>Kent, J.</dc:creator>
<dc:creator>Okafor, A.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Parameswaran, P.</dc:creator>
<dc:creator>Bustamante, E.</dc:creator>
<dc:creator>Ramani, B.</dc:creator>
<dc:creator>Diao, Y.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Soderblom, E. J.</dc:creator>
<dc:creator>Soderling, S. H.</dc:creator>
<dc:date>2026-01-17</dc:date>
<dc:identifier>doi:10.64898/2026.01.16.699530</dc:identifier>
<dc:title><![CDATA[In vivo Proximity & Spatial Proteomics with CRISPR Screening Identify STXBP1 as a Protective Modifier of a-synuclein Toxicity in Dopamine Neurons.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.16.699952v1?rss=1">
<title>
<![CDATA[
Rigor and Transparency in two neurotrauma-publishing journals: editorial policies improve transparent reporting. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.16.699952v1?rss=1</link>
<description><![CDATA[
Preclinical research in traumatic brain injury (TBI) continues to significantly increase knowledge and yield a large number of peer-reviewed studies, but translation of these results to the clinical setting has been minimal. Rigor and transparency factors such as concealment of group allocation (e.g., "blinding) or ensuring that reagents are identifiable are critical in ensuring that scientific studies are replicable and translatable. Yet, nearly all efforts aimed at measuring these factors have concluded that reporting practices are problematic and incomplete. One way to improve transparency of reporting practices is to require that authors address a set of transparency related items in some way, such as a checklist or a paper section. Recently, Journal of Neurotrauma, a leading publisher of preclinical TBI research, instituted a required rigor-related section, which is explained to authors via a set of transparency, rigor, and reproducibility (TRR) instructions (one example for each manuscript type). These documents include specific transparency sections explaining blinding, power calculations, protocols, code, and data deposition. Experimental Neurology is a journal that is similar in size, impact and topic but the journal does not have explicit instructions to authors about transparency items. The purpose of this study was to assess the degree to which transparency reporting items were included in published manuscripts comparing reporting practices in the Journal of Neurotrauma and Experimental Neurology. We used a commercial software, SciScore, which is an AI tool tuned to detect rigor/transparency sentences in published manuscripts and count the number found (roughly dividing by the number expected) to obtain a score. Overall, SciScore found that in 6 of 8 items that were explicitly asked for, such as power calculations, investigator blinding, inclusion criteria, attrition, and data were significantly greater (more than 10%) compared to Experimental Neurology. However in Journal of Neurotrauma papers with the extra rigor section, 3 of 4 rigor items that were not explicitly asked for in the template rigor documents, such as subject demographics or transparent antibody reporting were not different from Experimental Neurology. One item, reporting of the sex of subjects was significantly better in Experimental Neurology. This shows that the Journal of Neurotrauma required rigor section is effective in improving reporting, but it would be far better if sex as a biological variable and transparent reporting of reagents (items present on major checklists including NIH rigor criteria) would be included.
]]></description>
<dc:creator>Bandrowski, A. E.</dc:creator>
<dc:creator>Namburi, A.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Floyd, C. L.</dc:creator>
<dc:creator>Martone, M. E.</dc:creator>
<dc:creator>PRECISE-TBI Authors, T.</dc:creator>
<dc:date>2026-01-19</dc:date>
<dc:identifier>doi:10.64898/2026.01.16.699952</dc:identifier>
<dc:title><![CDATA[Rigor and Transparency in two neurotrauma-publishing journals: editorial policies improve transparent reporting.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.18.700232v1?rss=1">
<title>
<![CDATA[
Cancer drives atherosclerotic plaque vulnerability by inducing pathological angiogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.18.700232v1?rss=1</link>
<description><![CDATA[
Cardiovascular disease (CVD) is the leading cause of death for many cancer survivors, a phenomenon traditionally attributed to shared risk factors and cardiotoxic chemotherapies. Here, we hypothesized that cancer may also directly promote atherosclerosis. Propensity-matched analyses confirmed significantly elevated cardiovascular event rates amongst cancer patients, independent of comorbidities such as smoking. Atheroprone mice implanted with colorectal tumors demonstrated accelerated features of plaque vulnerability, driven by pathological angiogenesis and intraplaque hemorrhage. Mechanistically, tumor-secreted TNF- induced the pro-angiogenic factor LRG1 across multiple murine models and human plaques. Therapeutic interventions targeting these pathways, including with FDA-approved cytokine inhibitors or tumor resection, prevented plaque destabilization in mice and reduced coronary revascularization rates in patients. Together, these findings suggest that cancer may causally promote CVD and unveil novel translational strategies for cancer survivors.
]]></description>
<dc:creator>Luo, L.</dc:creator>
<dc:creator>Fu, C.</dc:creator>
<dc:creator>Jarr, K.-U.</dc:creator>
<dc:creator>Baylis, R.</dc:creator>
<dc:creator>Sun, V. H.</dc:creator>
<dc:creator>Heemelaar, J.</dc:creator>
<dc:creator>von Scheidt, M.</dc:creator>
<dc:creator>Ramirez, D.</dc:creator>
<dc:creator>Krefting, J.</dc:creator>
<dc:creator>Sachs, N.</dc:creator>
<dc:creator>Wettich, J. L.</dc:creator>
<dc:creator>Winter, H.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Adkar, S.</dc:creator>
<dc:creator>Haas, A.</dc:creator>
<dc:creator>Gonzalez, M.</dc:creator>
<dc:creator>Nead, K. T.</dc:creator>
<dc:creator>Maegdefessel, L.</dc:creator>
<dc:creator>Schunkert, H.</dc:creator>
<dc:creator>Neilan, T. G.</dc:creator>
<dc:creator>Leeper, N. J.</dc:creator>
<dc:date>2026-01-20</dc:date>
<dc:identifier>doi:10.64898/2026.01.18.700232</dc:identifier>
<dc:title><![CDATA[Cancer drives atherosclerotic plaque vulnerability by inducing pathological angiogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.19.700410v1?rss=1">
<title>
<![CDATA[
Selective Immune Silencing by Targeted TGF-β Agonists 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.19.700410v1?rss=1</link>
<description><![CDATA[
Depletion of pathogenic T and B cells is a pillar of first-line therapies for inflammatory, autoimmune, and transplantation-related immunological diseases. However, concerns about adverse events, safety in immunocompromised patients, and disease relapse from incomplete depletion, limit clinical utility. Here, we exploit the immunosuppressive properties of Transforming growth factor beta (TGF-{beta}), through selective "silencing" of T and B cells by a targeted TGF-{beta} mimic agonist derived from Helminths. CD4 and CD8 T cell-targeted TGF-{beta} agonists effectively silence antigen-stimulated T cell activation and expansion in mice and human spleen organoids. A mouse CD4 T cell-targeted TGF-{beta} agonist silences antigen-specific antibody responses by reprogramming pro-inflammatory Th1 and T follicular helper cells into quiescent or regulatory T cell phenotypes in vivo. A human CD19 B cell-targeted TGF-{beta} agonist silences antibody responses by disrupting differentiation of germinal center B cells into antibody-secreting cells in human spleen organoids. Cell-type-specific targeted TGF-{beta} agonists ameliorate disease activity in multiple mouse models with minimal off-target effects in vivo. Thus, cell-selective TGF-{beta} agonism is a versatile therapeutic strategy for precise silencing of immune functions.
]]></description>
<dc:creator>Sun, Q.</dc:creator>
<dc:creator>Ogishi, M.</dc:creator>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Barrett, A. K.</dc:creator>
<dc:creator>Yan, H.</dc:creator>
<dc:creator>Sola, E.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Xiao, P.</dc:creator>
<dc:creator>Lyu, H.</dc:creator>
<dc:creator>Salehi, A.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:creator>Lanz, T. V.</dc:creator>
<dc:creator>Davis, M. M.</dc:creator>
<dc:creator>Negrin, R. S.</dc:creator>
<dc:creator>Garcia, K. C.</dc:creator>
<dc:date>2026-01-20</dc:date>
<dc:identifier>doi:10.64898/2026.01.19.700410</dc:identifier>
<dc:title><![CDATA[Selective Immune Silencing by Targeted TGF-β Agonists]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.17.700100v1?rss=1">
<title>
<![CDATA[
Memory erasure by dopamine-gated retrospective learning 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.17.700100v1?rss=1</link>
<description><![CDATA[
Erasing outdated memories is crucial for adaptive behavior. Yet once a cue-outcome association is learned, repeated cue exposure without outcome suppresses conditioned behavior without erasing the underlying memory. This allows rapid behavioral recovery when outcomes are reintroduced. Here, we confirm this limitation for standard "prospective extinction" protocols that present cues without the associated outcome, but show that true memory erasure is achieved by inverting the paradigm: presenting outcomes without associated cues, i.e., "retrospective extinction". We demonstrate that orbitofrontal cortex activity at outcome is necessary for the rapid behavioral recovery following prospective extinction, and that mesolimbic dopamine activity at outcome is necessary for retrospective extinction. These findings reconceptualize extinction mechanisms and suggest complementary strategies to mitigate relapse and erase maladaptive memories.
]]></description>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Zsembik, L.</dc:creator>
<dc:creator>Farouq, F.</dc:creator>
<dc:creator>Chakraborty, R.</dc:creator>
<dc:creator>Belur, N.</dc:creator>
<dc:creator>Zhou, M.</dc:creator>
<dc:creator>Sanders, A. D.</dc:creator>
<dc:creator>Wang, S. X.</dc:creator>
<dc:creator>Srinivasan, A.</dc:creator>
<dc:creator>Cox, S. M. L.</dc:creator>
<dc:creator>Garr, E.</dc:creator>
<dc:creator>Brooke, S.</dc:creator>
<dc:creator>Janak, P. H.</dc:creator>
<dc:creator>Leyton, M.</dc:creator>
<dc:creator>Chen, R.</dc:creator>
<dc:creator>Namboodiri, V. M. K.</dc:creator>
<dc:date>2026-01-21</dc:date>
<dc:identifier>doi:10.64898/2026.01.17.700100</dc:identifier>
<dc:title><![CDATA[Memory erasure by dopamine-gated retrospective learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.21.700886v1?rss=1">
<title>
<![CDATA[
Sequence and Structural Alignments Reveal Insights into ANKLE2 Evolution and Function 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.21.700886v1?rss=1</link>
<description><![CDATA[
ANKLE2 is an enigmatic protein with emerging roles in cell division, development, and virus replication. While ANKLE2 orthologs are present in all animals, its domain composition has evolved over time. ANKLE2s two namesake domains, the ankyrin repeat and LEM domains, have clear and defined roles; however nearly all ANKLE2 orthologs have at least three other structured domains with poorly understood purposes. In this study, we performed sequence and structural alignments of ANKLE2 orthologs to improve our understanding of the proteins evolution and function. We identified that ANKLE2s transmembrane domain likely evolved more recently and coincided with loss of VAPA interaction as a membrane anchoring mechanism. We show that despite stark differences in amino acid sequence, the structure of the LEM and ankyrin repeat domains are highly conserved across ANKLE2 orthologs. To investigate ANKLE2s uncharacterized domains, we performed structural alignments to identify similar proteins. This revealed surprising similarities between portions of ANKLE2 and nuclease or nucleic acid-binding proteins. However, ANKLE2 lacks key motifs imparting function in these domains, which was confirmed by experimental interrogation. We further identified that loss of ANKLE2 is correlated with changes in DNA damage response and micronuclei formation. We believe this methodology demonstrates the power of combining structural predictions with classical molecular techniques in exploring poorly understood proteins.

ImportanceANKLE2 is a scaffolding protein present in all animals; however much of its function is poorly understood. By evaluating ANKLE2 sequence and structure from many different organisms and comparing its various domains with other proteins, we gain insight into how ANKLE2 evolved and what cellular roles it might be fulfilling. Further, this approach can be used to investigate other understudied or uncharacterized proteins.
]]></description>
<dc:creator>Fishburn, A. T.</dc:creator>
<dc:creator>Florio, C. J.</dc:creator>
<dc:creator>Skawinski, C. L. S.</dc:creator>
<dc:creator>Becker, S. S.</dc:creator>
<dc:creator>Holleman, E.</dc:creator>
<dc:creator>Robertson, A. E.</dc:creator>
<dc:creator>Sitchon, R.</dc:creator>
<dc:creator>Chedin, F.</dc:creator>
<dc:creator>Shah, P. S.</dc:creator>
<dc:date>2026-01-21</dc:date>
<dc:identifier>doi:10.64898/2026.01.21.700886</dc:identifier>
<dc:title><![CDATA[Sequence and Structural Alignments Reveal Insights into ANKLE2 Evolution and Function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.19.700188v1?rss=1">
<title>
<![CDATA[
MicroRNA regulatory targets of VO2peak in older adult participants of the Study of Muscle Mobility and Aging (SOMMA) 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.19.700188v1?rss=1</link>
<description><![CDATA[
Skeletal muscle aging (sarcopenia) is associated with reduced peak oxygen consumption (VO peak) during exercise, a key determinant of physical function and overall health. However, the molecular mechanisms linking muscle aging to low VO peak remain poorly understood. We aimed to identify miRNA signatures and miRNA-gene regulatory networks associated with VO peak in older adults. Using small RNA and mRNA sequencing, we analyzed skeletal muscle from 72 SOMMA participants (70-79 years old) with low or high VO peak (n = 18/group) and from 36 participants spanning the full VO peak spectrum. Differential expression was assessed using LIMMA, with pathway and network analyses performed using Ingenuity Pathway Analysis (IPA) and Weighted Gene Co-expression Network Analysis (WGCNA). We detected 1,408 miRNAs and 16,210 genes; among these, 14 miRNAs and 2,018 genes were differentially expressed (FDR < 0.05). The 14 miRNAs regulated 142 genes, and expression of 10 miRNAs inversely correlated with 50 genes enriched in mitochondrial, sirtuin-1, and nitric oxide signaling pathways. Regression analyses identified 21 miRNAs and 1,744 genes significantly correlated with VO peak after adjusting for age and sex. WGCNA revealed 10 co-expression modules associated with VO peak, with the cyan module showing the strongest correlation and enrichment for nitric oxide signaling genes. These findings highlight novel miRNA-mediated molecular pathways potentially contributing to low VO peak and skeletal muscle aging in older adults. Future studies will further investigate these miRNA-gene interactions to uncover therapeutic targets for preserving muscle function with age.
]]></description>
<dc:creator>Karere, G.</dc:creator>
<dc:creator>Hsu, F.-C.</dc:creator>
<dc:creator>Hepple, R. T.</dc:creator>
<dc:creator>Coen, P. M.</dc:creator>
<dc:creator>Cummings, S. R.</dc:creator>
<dc:creator>Newman, A. B.</dc:creator>
<dc:creator>Glynn, N. W.</dc:creator>
<dc:creator>Sparks, L. M.</dc:creator>
<dc:creator>Lane, N. E.</dc:creator>
<dc:creator>Hayward, A. G.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Wagner, N.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Chan, J.</dc:creator>
<dc:creator>Cox, L.</dc:creator>
<dc:creator>Kritchevsky, S. B.</dc:creator>
<dc:date>2026-01-22</dc:date>
<dc:identifier>doi:10.64898/2026.01.19.700188</dc:identifier>
<dc:title><![CDATA[MicroRNA regulatory targets of VO2peak in older adult participants of the Study of Muscle Mobility and Aging (SOMMA)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.20.700462v1?rss=1">
<title>
<![CDATA[
The role of cohesin loading at enhancers in the flux of loop extrusion and long-range transcriptional control 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.20.700462v1?rss=1</link>
<description><![CDATA[
Enhancers have been proposed to act as privileged loading sites for cohesin, raising the idea that they actively fold the genome to engage distal target promoters for transcription. Supporting this idea, NIPBL/MAU2, which is required for cohesin loading, binds at enhancers in mouse embryonic stem cells. However, we find that driving cohesin recruitment near an enhancer strongly inhibits transcription from its target distal promoter, indicating that strong focal cohesin loading at enhancers is not compatible with their long-range regulatory functions. Quantitative experiments and biophysical modeling further indicate that cohesin loading at enhancers does not make major contributions to genome-wide cohesin binding and chromosome folding patterns. Instead, cohesin must load throughout the genome to extrude it, regardless of enhancer proximity, with the major determinants of cohesin traffic being extrusion barriers such as transcription and clustered CTCF sites. These findings indicate that enhancer function is largely ancillary to the general mechanisms of chromosome folding, informing further study of the relationship between genome architecture and transcriptional regulation.
]]></description>
<dc:creator>Anderson, E. C.</dc:creator>
<dc:creator>Rahmaninejad, H.</dc:creator>
<dc:creator>Aljahani, A.</dc:creator>
<dc:creator>Arnold, E. M.</dc:creator>
<dc:creator>Adachi, A. S.</dc:creator>
<dc:creator>Shah, R.</dc:creator>
<dc:creator>Hansen, K. L.</dc:creator>
<dc:creator>Cavka, I.</dc:creator>
<dc:creator>Boettiger, A. N.</dc:creator>
<dc:creator>Fudenberg, G.</dc:creator>
<dc:creator>Nora, E. P.</dc:creator>
<dc:date>2026-01-22</dc:date>
<dc:identifier>doi:10.64898/2026.01.20.700462</dc:identifier>
<dc:title><![CDATA[The role of cohesin loading at enhancers in the flux of loop extrusion and long-range transcriptional control]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.21.700667v1?rss=1">
<title>
<![CDATA[
Histone 3 lysine 9 dimethylation by the G9a-GLP heterodimer requires intranucleosomal product reading 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.21.700667v1?rss=1</link>
<description><![CDATA[
Repressive histone methyltransferases carry a catalytic ("write") domain and a separate domain specialized for recognizing ("reading") the reaction product. This read-write configuration acts as a positive feedback mechanism for epigenetic maintenance and the growth of repressive chromatin domains. Feedback exhibits as catalytic stimulation and is understood to act towards a proximal (trans) nucleosome. Whether this stimulation affects a specific methylation transition and whether it is restricted to trans-stimulation remains opaque. Here, we dissect the positive feedback in the heterodimeric histone 3 lysine 9 (H3K9) mono- and dimethlyase G9a-GLP, which carries two catalytic SET and two product-reading Ankyrin repeat (ANK) domains. We find that reading by both ANK domains is required for H3K9 di-, but not monomethylation on nucleosomes and for tight binding to them. As this read-writing occurs on dilute mononucleosomes, we propose that intranucleosomal feedback occurs for G9a-GLP. Swapping the ANK domains results in loss of dimethylation while maintaining nucleosome binding, indicating catalytic coupling of nucleosome methylation intermediates to reading. Crosslinking mass spectrometry reveals specific G9a surfaces that contact nucleosomal methylation intermediates. Structural approaches reveal how these surfaces position the G9a ANK domain on the methylation-intermediate nucleosome and stabilize G9a-GLP on chromatin during the reaction.
]]></description>
<dc:creator>Yousefi, F.</dc:creator>
<dc:creator>Simental, E. A.</dc:creator>
<dc:creator>Du, Y.</dc:creator>
<dc:creator>Whedon, S. D.</dc:creator>
<dc:creator>Trnka, M. J.</dc:creator>
<dc:creator>Darling, D.</dc:creator>
<dc:creator>Jia, S.</dc:creator>
<dc:creator>Panning, B.</dc:creator>
<dc:creator>Cole, P. A.</dc:creator>
<dc:creator>Halic, M.</dc:creator>
<dc:creator>Al-Sady, B.</dc:creator>
<dc:date>2026-01-22</dc:date>
<dc:identifier>doi:10.64898/2026.01.21.700667</dc:identifier>
<dc:title><![CDATA[Histone 3 lysine 9 dimethylation by the G9a-GLP heterodimer requires intranucleosomal product reading]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.22.701024v1?rss=1">
<title>
<![CDATA[
Conserved assembly architecture of the essential herpesvirus packaging accessory factor 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.22.701024v1?rss=1</link>
<description><![CDATA[
To create a new wave of infectious virions, all herpesviruses require an accessory factor of unknown function to package their viral genomes into nascent capsids. Here, we present cryo-EM structures of the packaging accessory factor from the -herpesvirus herpes simplex virus type 1 (HSV-1, UL32) and the {beta}-herpesvirus human cytomegalovirus (HCMV, UL52). Unlike homologs from the {gamma}-herpesviruses, neither UL32 nor UL52 form stable homopentameric rings. UL52 forms incomplete pentameric rings lacking one or two protomers. UL32 does not form stable higher-order species, but stabilization through chemical crosslinking revealed a novel quaternary structure where three pentameric rings assemble into a "tripentamer." Our results reveal that herpesvirus packaging accessory factors adopt distinct oligomeric states but are constrained to pentameric symmetry. Assembly of protomers into a ring creates a positively charged central channel that we show is critical for infectious virus production in HSV-1. Taken together, our study points to a structurally conserved, essential function of packaging accessory factors across the Herpesviridae.
]]></description>
<dc:creator>Bailey, E. J.</dc:creator>
<dc:creator>Devarkar, S. C.</dc:creator>
<dc:creator>Szczepaniak, R.</dc:creator>
<dc:creator>Meissner, L. M.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Wu, C.</dc:creator>
<dc:creator>Weller, S. K.</dc:creator>
<dc:creator>Xiong, Y.</dc:creator>
<dc:creator>Didychuk, A. L.</dc:creator>
<dc:date>2026-01-22</dc:date>
<dc:identifier>doi:10.64898/2026.01.22.701024</dc:identifier>
<dc:title><![CDATA[Conserved assembly architecture of the essential herpesvirus packaging accessory factor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.20.700519v1?rss=1">
<title>
<![CDATA[
Federated single-cell QTL meta-analysis reveals novel disease mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.20.700519v1?rss=1</link>
<description><![CDATA[
Genetic effects on gene expression are often cell type-specific and obscured in bulk analyses. To resolve this context-dependent regulation, we performed a federated cis-eQTL meta-analysis across 12 PBMC datasets (2,032 individuals, 2.5 million cells). Across six immune cell types, we identified cis-eQTLs for 6,592 genes and fine-mapped 14,985 independent loci. Notably, the 42% of eQTLs that were undetected in a bulk eQTL study on 43,301 whole blood samples also showed stronger enrichment for disease GWAS loci. We further identified three genome-wide significant and 65 suggestive loci affecting the abundance of (rare) immune cell types and validated these using previously reported hematological GWAS and bulk-derived trans-eQTLs. Integrating single-cell cis-eQTLs with bulk trans-eQTLs enabled us to anchor 6,382 trans-eGenes (37.2% novel) to upstream regulators and reconstruct directed gene regulatory relationships. For example, a hemorrhoidal disease-associated variant showed a CD4+ T cell-specific cis-eQTL on BACH1 that colocalized with 45 immune and metabolic trans-eGenes. These results demonstrate the power of single-cell QTL meta-analysis in interpreting complex trait genetics.
]]></description>
<dc:creator>Kaptijn, D.</dc:creator>
<dc:creator>Michielsen, L.</dc:creator>
<dc:creator>Neavin, D.</dc:creator>
<dc:creator>Ripoll-Cladellas, A.</dc:creator>
<dc:creator>Alquicira-Hernandez, J. E.</dc:creator>
<dc:creator>Korshevniuk, M.</dc:creator>
<dc:creator>Lee, J. T. H.</dc:creator>
<dc:creator>Oelen, R.</dc:creator>
<dc:creator>Vochteloo, M.</dc:creator>
<dc:creator>Warmerdam, R.</dc:creator>
<dc:creator>Ando, Y.</dc:creator>
<dc:creator>Ban, M.</dc:creator>
<dc:creator>Bayaraa, O.</dc:creator>
<dc:creator>Berg, M.</dc:creator>
<dc:creator>van Blokland, I.</dc:creator>
<dc:creator>Considine, D.</dc:creator>
<dc:creator>Dieng, M. M.</dc:creator>
<dc:creator>Edahiro, R.</dc:creator>
<dc:creator>Gordon, M. G.</dc:creator>
<dc:creator>Groot, H. E.</dc:creator>
<dc:creator>van der Harst, P.</dc:creator>
<dc:creator>Heinig, M.</dc:creator>
<dc:creator>Hon, C.-C.</dc:creator>
<dc:creator>Idaghdour, Y.</dc:creator>
<dc:creator>Kathail, P.</dc:creator>
<dc:creator>de Klein, N.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Losert, C.</dc:creator>
<dc:creator>Manikanda, V.</dc:creator>
<dc:creator>Moody, J.</dc:creator>
<dc:creator>Naeem, H.</dc:creator>
<dc:creator>Mokrab, Y.</dc:creator>
<dc:creator>Nawijn, M. C.</dc:creator>
<dc:creator>Netea, M.</dc:creator>
<dc:creator>Niewold, J.</dc:creator>
<dc:creator>Okada, Y.</dc:creator>
<dc:creator>Sawcer, S.</dc:creator>
<dc:creator>Soulama, I.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Tsepilov, Y.</dc:creator>
<dc:creator>Park, W.-Y.</dc:creator>
<dc:creator>Rajagopalan, D.</dc:creator>
<dc:creator>Shahin, T.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2026-01-23</dc:date>
<dc:identifier>doi:10.64898/2026.01.20.700519</dc:identifier>
<dc:title><![CDATA[Federated single-cell QTL meta-analysis reveals novel disease mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.21.700898v1?rss=1">
<title>
<![CDATA[
Biophysically realistic network-level transport model of tau progression with exosome-mediated release and uptake processes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.21.700898v1?rss=1</link>
<description><![CDATA[
The spatiotemporal progression of tau aggregates in neurodegenerative diseases like Alzheimers follows the brains structural connectome, yet a profound gap exists between the slow macroscopic spread observed over years and the rapid protein kinetics occurring over hours. Current graph-diffusion models fail to reconcile this timescale disparity or incorporate the cellular mechanisms driving transmission. Here, we advance the Network Transport Model (NTM) to bridge these scales by integrating directional active transport along microtubules, continuous toxic tau production, and exosome-mediated trans-neuronal release and uptake. This framework constitutes one of the most mechanistically complete and biologically detailed models of tau spread on the whole brain to date, representing a significant innovation in how multiscale proteinopathies are simulated. To overcome the computational complexity of the underlying partial differential equations, we developed a quasi-static approximation that separates fast axonal transport from slow network-wide exchange.

Simulations on the whole-brain mouse connectome demonstrate that this framework emergently replicates empirical tau propagation patterns without the need for case-specific empirical fitting. Our results identify trans-neuronal release and uptake rates as the primary mechanistic "bottleneck" on macroscopic spread, providing a biologically grounded explanation for the diseases slow progression. Furthermore, we find that high aggregation rates can paradoxically sequester tau within neurons, limiting global transmission, while transport polarity (anterograde vs. retrograde) fundamentally dictates spatial patterning. By linking molecular mechanics to system-wide pathology, this model provides a predictive "in-silico" framework to evaluate how cellular-targeted interventions might alter the trajectory of tauopathic dementias.
]]></description>
<dc:creator>Barron, N.</dc:creator>
<dc:creator>Tora, V.</dc:creator>
<dc:creator>Cozzolino, E.</dc:creator>
<dc:creator>Bertsch, M.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:date>2026-01-23</dc:date>
<dc:identifier>doi:10.64898/2026.01.21.700898</dc:identifier>
<dc:title><![CDATA[Biophysically realistic network-level transport model of tau progression with exosome-mediated release and uptake processes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.22.700386v1?rss=1">
<title>
<![CDATA[
Heterologous betacoronavirus spike immunization in nonhuman primates elicits cross-reactive antibodies that neutralize both sarbeco- and merbecoviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.22.700386v1?rss=1</link>
<description><![CDATA[
In anticipation of future coronavirus (CoV) pandemics, developing vaccines that elicit broadly neutralizing antibodies (bnAbs) against diverse CoVs is critical. Here, we vaccinated rhesus macaques with SARS-CoV-2 spike (S)-protein, then boosted with heterologous {beta}-CoV S-proteins to focus responses to common conserved S2 bnAb epitopes. Initial SARS-CoV-2 priming elicited receptor-binding domain (RBD)-focused responses, while MERS-CoV boosting redirected responses toward the S2 region, including the stem-helix bnAb site. Although S2-directed serum cross-neutralization was undetectable and most isolated cross-reactive monoclonal antibodies (mAbs) targeted non-neutralizing epitopes, two S2 stem-helix mAbs were identified from memory B cells. These bnAbs neutralized diverse sarbeco- and merbecoviruses, including MERS-CoV, and conferred robust in vivo protection against SARS-CoV-2 challenge. Structural studies revealed that these macaque bnAbs closely mimic human S2-stem bnAbs induced by infection. These findings provide proof-of-principle for vaccination strategies that elicit broadly protective {beta}-coronavirus responses and highlight non-human primates as a translational model for evaluating S2-targeted immunogens.
]]></description>
<dc:creator>Dueker, K.</dc:creator>
<dc:creator>Capozzola, T.</dc:creator>
<dc:creator>Feng, Z.</dc:creator>
<dc:creator>Lin, R. N.</dc:creator>
<dc:creator>Hurtado, J.</dc:creator>
<dc:creator>Bangaru, S.</dc:creator>
<dc:creator>Yuan, M.</dc:creator>
<dc:creator>Beutler, N.</dc:creator>
<dc:creator>Garcia, E.</dc:creator>
<dc:creator>He, W.-t.</dc:creator>
<dc:creator>Callaghan, S.</dc:creator>
<dc:creator>Avillion, G.</dc:creator>
<dc:creator>Vo, L.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Torres, J. L.</dc:creator>
<dc:creator>Musharrafieh, R.</dc:creator>
<dc:creator>Song, G.</dc:creator>
<dc:creator>Mishra, N.</dc:creator>
<dc:creator>Sharma, P.</dc:creator>
<dc:creator>Yong, P.</dc:creator>
<dc:creator>Anzanello, F.</dc:creator>
<dc:creator>Kaczmarek-Michaels, K.</dc:creator>
<dc:creator>Ben-Akiva, E.</dc:creator>
<dc:creator>Silva, M.</dc:creator>
<dc:creator>Melo, M.</dc:creator>
<dc:creator>Makhdoomi, M.</dc:creator>
<dc:creator>Westfall-Gomez, E.</dc:creator>
<dc:creator>Rinaldi, W.</dc:creator>
<dc:creator>Ferguson, M.</dc:creator>
<dc:creator>Safonova, Y.</dc:creator>
<dc:creator>Crotty, S.</dc:creator>
<dc:creator>Irvine, D. J.</dc:creator>
<dc:creator>Rogers, T.</dc:creator>
<dc:creator>Ward, A. B.</dc:creator>
<dc:creator>Briney, B.</dc:creator>
<dc:creator>Wilson, I. A.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>andrabi, R.</dc:creator>
<dc:date>2026-01-23</dc:date>
<dc:identifier>doi:10.64898/2026.01.22.700386</dc:identifier>
<dc:title><![CDATA[Heterologous betacoronavirus spike immunization in nonhuman primates elicits cross-reactive antibodies that neutralize both sarbeco- and merbecoviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.22.701150v1?rss=1">
<title>
<![CDATA[
Synthetic Hybrid Receptors for Safer and Programmable T Cell Therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.22.701150v1?rss=1</link>
<description><![CDATA[
Engineered T cell therapies have achieved significant clinical success in hematological malignancies but remain largely ineffective in solid tumors. Overcoming this limitation requires strategies that enhance T cell function while avoiding systemic immune toxicities and pathological T cell states. Existing approaches typically rely on constitutive gene overexpression or suppression to augment potency or remodel the tumor microenvironment, but these strategies frequently lead to dysregulated immune activation and dose-limiting toxicity. Here, we present Hybrid Receptors (Hybrid-Rs), a modular receptor platform that integrates features of chimeric antigen receptors (CARs) and SyNthetic Intramembrane Proteolysis Receptors (SNIPRs) to couple antigen-dependent T cell activation with programmable gene regulation. Hybrid-Rs enable precise, context-dependent control of T cell potency, differentiation states, and conditional expression of secreted immunotherapeutic payloads with otherwise prohibitive toxicity. Hybrid-Rs are readily humanized and compatible with precision genome editing in primary human T cells, providing a direct and practical path to clinical translation.
]]></description>
<dc:creator>Foisey, M.</dc:creator>
<dc:creator>Garcia, J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Hilburger, C.</dc:creator>
<dc:creator>Thienpont, C.</dc:creator>
<dc:creator>Chaves-Martinez, A.</dc:creator>
<dc:creator>Belkaya, S.</dc:creator>
<dc:creator>Truong, T.</dc:creator>
<dc:creator>Zhu, I.</dc:creator>
<dc:creator>Liu, R.</dc:creator>
<dc:creator>Hyrenius-wittsten, A.</dc:creator>
<dc:creator>Almeida, R.</dc:creator>
<dc:creator>Shy, B. R.</dc:creator>
<dc:creator>Allen, G. M.</dc:creator>
<dc:creator>Wyman, S.</dc:creator>
<dc:creator>Roybal, K. T.</dc:creator>
<dc:date>2026-01-23</dc:date>
<dc:identifier>doi:10.64898/2026.01.22.701150</dc:identifier>
<dc:title><![CDATA[Synthetic Hybrid Receptors for Safer and Programmable T Cell Therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.23.701345v1?rss=1">
<title>
<![CDATA[
Spatially resolved transcriptome-metabolome integration reveals region-specific glial lipid dysregulation associated with Alzheimer's pathology 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.23.701345v1?rss=1</link>
<description><![CDATA[
Glial cells maintain the brains lipid and energy balance, and their breakdown is increasingly recognized as a causal contributor to Alzheimers disease (AD). While this concept is established, no approach has directly shown how glial homeostatic failure manifests across brain regions and microenvironments or how it links local pathology, such as plaques, to global metabolic imbalance. To address this gap, we developed iMIST, an integrated platform that combines MALDI-based metabolite imaging, histology, and spatial transcriptomics within a single tissue section to align molecular and anatomical information. Using a mouse model of late-onset AD that recapitulates both amyloid deposition and metabolic vulnerability, iMIST revealed that glial lipid dysregulation is widespread but spatially specialized. In gray matter, plaque-associated microglia were associated with upregulated glycerophospholipid-remodeling in cortico-thalamic areas indicating metabolic stress around local pathology. In contrast, white matter tracts rich in lipid-producing oligodendrocytes show plaque-independent deficits in galactosylceramide metabolism reflecting their high myelin demand. Both processes intensify with age, transforming adaptive glial responses into persistent metabolic dysfunction. Together, these findings demonstrate the spatial interplay between global glial metabolic imbalance and local microenvironmental stressors associated with AD pathology. By integrating transcriptomic and metabolomic information in situ, iMIST provides a framework for uncovering how regional glial vulnerability shapes the pathogenesis of neurodegenerative diseases.
]]></description>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Leon, J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Oehler, A.</dc:creator>
<dc:creator>Modavi, C.</dc:creator>
<dc:creator>Abate, A. R.</dc:creator>
<dc:creator>Condello, C.</dc:creator>
<dc:date>2026-01-25</dc:date>
<dc:identifier>doi:10.64898/2026.01.23.701345</dc:identifier>
<dc:title><![CDATA[Spatially resolved transcriptome-metabolome integration reveals region-specific glial lipid dysregulation associated with Alzheimer's pathology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.23.701436v1?rss=1">
<title>
<![CDATA[
Dynamic reorganization of Developmental to Adult Genome Topology Controls the Initiation and Stabilization of the Human Muscle Stem Cell State 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.23.701436v1?rss=1</link>
<description><![CDATA[
Developmental gene expression is under tight temporal and spatial control. This regulation is imparted by tissue specific enhancers that integrate developmental signals into transcriptional responses to allow for developmental progression. Often species specific, the enhancers that regulate human muscle progenitor and stem cell gene expression are currently unknown. Here, we define the 3D chromatin organization of human muscle development and reveal key changes across the human genome that are associated with multiple layers of 3D genome reorganization during the transition from a more progenitor-like to muscle stem cell state, including a reduction of TAD numbers and an increase in CTCF binding at TAD boundaries and chromatin loops throughout developmental progression. Specifically, we found that increased CTCF occupancy at human enhancers of PAX7 in stem cells holds enhancer-promoter (e-p) loops for timely activation of PAX7 enhancers during early human development. These findings demonstrate that stem cell state acquisition is stabilized earlier than previously known and provide unprecedented insights into the initiation and control of the muscle stem cell state in humans.
]]></description>
<dc:creator>Romero, M. A.</dc:creator>
<dc:creator>Chien, P.</dc:creator>
<dc:creator>Nicoletti, C.</dc:creator>
<dc:creator>Liliom, H. L.</dc:creator>
<dc:creator>Cox, G.</dc:creator>
<dc:creator>Skuratovsky, E.</dc:creator>
<dc:creator>Saleh, K.</dc:creator>
<dc:creator>Gibbs, D.</dc:creator>
<dc:creator>Gane, L.</dc:creator>
<dc:creator>Hoang, D.-H.</dc:creator>
<dc:creator>Caputo, L.</dc:creator>
<dc:creator>Massenet, J.</dc:creator>
<dc:creator>Sobreira, D. R.</dc:creator>
<dc:creator>Puri, P. L.</dc:creator>
<dc:creator>Pyle, A. D.</dc:creator>
<dc:date>2026-01-25</dc:date>
<dc:identifier>doi:10.64898/2026.01.23.701436</dc:identifier>
<dc:title><![CDATA[Dynamic reorganization of Developmental to Adult Genome Topology Controls the Initiation and Stabilization of the Human Muscle Stem Cell State]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.24.701487v1?rss=1">
<title>
<![CDATA[
Cell-Autonomous and Systemic Circadian Regulation of Gene Expression in Adipocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.24.701487v1?rss=1</link>
<description><![CDATA[
Circadian clocks strongly influence adipocyte biology. Studying adipocyte circadian regulation in the absence of organismal cues isolates cell-autonomous clock control, providing insight into mechanisms relevant to metabolic disease. To resolve circadian core biological programs, we deeply sequenced bulk RNA from inguinal-derived, in vitro-differentiated adipocytes (IVDAs) over 2.5 days and compared clock-controlled genes (CCGs) with existing mouse supraclavicular brown (BAT) and epididymal white (eWAT) adipose tissue circadian time-course datasets. Using Phase Set Enrichment Analysis (PSEA), we report 21% of the protein-coding transcriptome is rhythmic in IVDAs. Intrinsic circadian regulation governs key processes in energy metabolism, molecular transport and transcription. Integration with in vivo datasets reveals that BAT and IVDAs exhibit more cohesive rhythmic pathways than does eWAT, clustering around the late-night early-morning transition. To explore how these pathways may be regulated, we reexamined a recent interscapular BAT cistrome dataset. Using IVDA transcription factors that were phase-aligned ([&le;]4hr) with in vivo as input, motif enrichment analysis revealed two temporally distinct regulatory programs; an early E-box activator ARNT-family/bHLH-PAS program was enriched for transcriptional regulation, RNA metabolism, and signaling pathways, and a late nuclear receptor-associated program enriched for energy metabolism, phospholipid biosynthesis, mitochondrial function, ECM organization, and nuclear receptor signaling. Overall, we identify novel rhythmic transcripts and define cell-autonomous circadian programs in adipocytes whose timing is further sculpted by systemic cues in vivo. Because obesity is associated with adipocyte hypertrophy and hyperplasia, processes likely influenced by circadian regulation, these findings advance our understanding of clock-controlled adipocyte metabolism and its contribution to metabolic dysfunction.
]]></description>
<dc:creator>Worthen, J. M.</dc:creator>
<dc:creator>Frederick, A. M.</dc:creator>
<dc:creator>Loros, J. J.</dc:creator>
<dc:creator>Dumesic, P. A.</dc:creator>
<dc:creator>Dunlap, J. C.</dc:creator>
<dc:date>2026-01-25</dc:date>
<dc:identifier>doi:10.64898/2026.01.24.701487</dc:identifier>
<dc:title><![CDATA[Cell-Autonomous and Systemic Circadian Regulation of Gene Expression in Adipocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.26.701664v1?rss=1">
<title>
<![CDATA[
The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.26.701664v1?rss=1</link>
<description><![CDATA[
Over the past decades, chikungunya virus (CHIKV), a re-emerging arthropod-borne alphavirus, has caused outbreaks in many (sub)tropical regions, but also in more temperate regions of the world, including Europe. CHIKV poses a significant health burden due to high infection rates during epidemics, symptom progression into chronic arthritic manifestations, and the lack of specific antiviral treatments. Multiple studies have shown that several viruses rely on host molecular chaperones, particularly the central Heat shock protein 70 (Hsp70), for replication. Hsp70s are guided by co-chaperones that drive their functionality and specificity. Here, we used chemical inhibitors of Hsp70-co-chaperone interactions to study the role of this molecular chaperone machinery in CHIKV replication. Our findings revealed that Hsp70 inhibition significantly reduces the CHIKV infectious particle production without affecting host-cell viability. Inhibition of the Hsp70-co-chaperone interaction primarily impedes the post-RNA replication stages of the CHIKV infectious cycle, affecting viral protein expression and reducing both the number and infectivity of released virions. Moreover, Hsp70 inhibition displayed antiviral activity in skin explants. Together, these results suggest that targeting the Hsp70 network could be a viable antiviral strategy against CHIKV infections.

Author summaryAs obligatory intracellular parasites, viruses rely entirely on the machinery of the host cell to produce new viral particles. In our study, we investigated whether a specific group of host proteins, known as molecular chaperones, is important for the replication of chikungunya virus (CHIKV), a reemerging mosquito-borne virus that can cause long-lasting joint pains and lacks specific antiviral treatments. We focused on one key family of molecular chaperones, called Heat shock protein 70s (Hsp70s), which supports protein folding and quality control in cells. We used chemical compounds to block Hsp70 function and observed that CHIKV replication was strongly reduced, while host cells remained healthy. We found that Hsp70 is especially important in the later stages of the virus life cycle, where it helps produce viral proteins and new infectious virus particles. When Hsp70 was blocked, fewer and less infectious virus particles were produced. We also showed that this effect holds true in biopsies of mouse skin tissue, which mimics the initial site of infection. These findings identify Hsp70 as an important host factor for CHIKV that may serve as a potential target for new antiviral therapies.
]]></description>
<dc:creator>van der Laan, M.</dc:creator>
<dc:creator>Verwimp, S.</dc:creator>
<dc:creator>Johnson, O. T.</dc:creator>
<dc:creator>Bouma, E. M.</dc:creator>
<dc:creator>Trappeniers, K.</dc:creator>
<dc:creator>Visscher, F. E.</dc:creator>
<dc:creator>van den Ende-Metselaar, H. H.</dc:creator>
<dc:creator>van de Pol, D. P. I.</dc:creator>
<dc:creator>Delang, L.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Kampinga, H. H.</dc:creator>
<dc:creator>Smit, J. M.</dc:creator>
<dc:date>2026-01-26</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701664</dc:identifier>
<dc:title><![CDATA[The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.26.701810v1?rss=1">
<title>
<![CDATA[
Genome-wide CRISPRi screen identifies basigin loss as protective in cardiac hypoxia 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.26.701810v1?rss=1</link>
<description><![CDATA[
Cardiac function depends on continuous oxidative metabolism, rendering cardiomyocytes highly vulnerable to oxygen deprivation. Here, we performed a genome-wide CRISPR interference (CRISPRi) screen in human iPSC-derived cardiomyocytes to identify genes that modulate survival during chronic hypoxia. This screen revealed that knockdown of basigin (BSG), a chaperone for the monocarboxylate transporters MCT1 and MCT4, confers robust protection. Canonically, hypoxic cells suppress pyruvate dehydrogenase (PDH) activity to reduce the oxidation of major fuel sources, thereby limiting TCA cycle flux, lowering oxygen consumption, and minimizing reactive oxygen species generated by an overly reduced electron transport chain (ETC). In contrast, we found that BSG inhibition reverses this response, prioritizing ATP maintenance during hypoxia and enhancing cardiomyocyte survival. Mechanistically, BSG loss restricts lactate efflux, leading to decreased PDH phosphorylation and increased glucose uptake for oxidation. Consistent with this, ETC subunits are more essential under hypoxia, highlighting cardiomyocytes unusual reliance on aerobic ATP production even when oxygen is limited. These findings challenge prevailing models of hypoxic adaptation by revealing cardiomyocyte-specific bioenergetic requirements and motivating future therapeutic efforts.
]]></description>
<dc:creator>Flanigan, W. R.</dc:creator>
<dc:creator>Midha, A. D.</dc:creator>
<dc:creator>Blume, S. Y.</dc:creator>
<dc:creator>Marti-Mateos, Y.</dc:creator>
<dc:creator>Costa, M. W.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Baik, A. H.</dc:creator>
<dc:creator>Huynh, H.</dc:creator>
<dc:creator>Susarla, G.</dc:creator>
<dc:creator>Bennett, N. K.</dc:creator>
<dc:creator>Nowak, R. A.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Jain, I. H.</dc:creator>
<dc:date>2026-01-27</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701810</dc:identifier>
<dc:title><![CDATA[Genome-wide CRISPRi screen identifies basigin loss as protective in cardiac hypoxia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.26.701898v1?rss=1">
<title>
<![CDATA[
Pre-myelinating oligodendrocyte ADGRG1 is required for axon ensheathment and CNS myelin formation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.26.701898v1?rss=1</link>
<description><![CDATA[
Myelin is essential for axonal health and rapid propagation of action potentials. In the central nervous system (CNS), myelination is initiated with axon ensheathment by pre-myelinating oligodendrocytes (preOLs), followed by iterative membrane wrapping and longitudinal extension along the axon. The molecular mechanisms that govern preOL development remain largely unknown. In this study, we identify the adhesion G protein-coupled receptor ADGRG1 (also called GPR56) as a key, evolutionarily conserved regulator of this process. We show that ADGRG1 is highly expressed in preOLs and that its conditional deletion in mouse preOLs leads to simplified preOL morphology and defective axon ensheathment, resulting in CNS hypomyelination. Live imaging in zebrafish demonstrated conserved function of Adgrg1 in axon ensheathment. Mechanistically, we show that ADGRG1 promotes preOL maturation by RhoA activation. Collectively, these findings reveal an ADGRG1-mediated RhoA signaling pathway that governs axon ensheathment and myelin formation.
]]></description>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Rodrigues, B. V.</dc:creator>
<dc:creator>Piller, M. V.</dc:creator>
<dc:creator>Call, C. L.</dc:creator>
<dc:creator>Mogha, A.</dc:creator>
<dc:creator>Belloso-Iguerategui, A.</dc:creator>
<dc:creator>Chiou, B.</dc:creator>
<dc:creator>Schmidt, R.</dc:creator>
<dc:creator>Wangzhou, A.</dc:creator>
<dc:creator>Thurber, A.</dc:creator>
<dc:creator>Dee, M.</dc:creator>
<dc:creator>Larson, C.</dc:creator>
<dc:creator>Monk, K.</dc:creator>
<dc:creator>Piao, X.</dc:creator>
<dc:date>2026-01-27</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701898</dc:identifier>
<dc:title><![CDATA[Pre-myelinating oligodendrocyte ADGRG1 is required for axon ensheathment and CNS myelin formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.28.699085v1?rss=1">
<title>
<![CDATA[
LANA-dependent Interaction of Host Factors DAXX and BRD4 Impact KSHV Lytic Replication 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.28.699085v1?rss=1</link>
<description><![CDATA[
Kaposis Sarcoma-Associated Herpesvirus (KSHV) is an oncogenic gammaherpesvirus that causes Kaposis Sarcoma (KS). KSHV alternates between latent and lytic phases. Latency is marked by limited viral gene expression and absence of virion production, whereas the lytic phase is characterized by the expression of all viral genes in a temporally and sequentially regulated cascade of immediate-early, early, and late genes, culminating in viral genome replication and infectious virion production. Both KSHV latent and lytic viral phases contribute to its pathogenesis and the development of KS. KSHV proteins hijack host proteins, rewiring host cellular processes and signaling pathways, and modulating host gene expression. KSHV establishes latency through stable interactions between viral proteins and host factors, with the Latency Associated Nuclear Antigen (LANA) serving as a key regulator that interacts with host machinery to maintain the viral episome and regulate viral replication. However, the consequences for host proteins binding viral proteins, including changes in their interaction networks and their impact on viral replication, remains poorly understood. Here, using Immunoprecipitation Mass Spectrometry (IP-MS), we identified and validated the host death domain-associated protein (DAXX) and DNA ligase 3 (LIG3) as major LANA-associated factors and discovered a LANA-dependent recruitment of Bromodomain-containing protein 4 (BRD4) to DAXX. This remodeling of the DAXX interactome guided functional analyses demonstrating roles for DAXX and BRD4 in supporting KSHV infection. Functional studies show that siRNA knockdown of DAXX or BRD4 in iSLK.BAC16 cells, followed by lytic reactivation, significantly induced viral gene transcription and protein expression of KSHV lytic genes ORF45, ORF59, ORF26, and K8.1. Conversely, siRNA knockdown of LIG3 reduced the transcription and protein expression of KSHV lytic genes. Viral genome replication and infectious virion production were elevated upon knockdown of DAXX or BRD4 and reduced after the knockdown of LIG3. Additionally, chemical inhibition of BRD4 activity by the drug JQ1 in iSLK.BAC16 cells, followed by lytic reactivation, resulted in elevated KSHV lytic gene expression, genome replication, and infectious virus production. Together, these data suggest that both DAXX and BRD4 host genes contribute to KSHV latency maintenance, while LIG3 is required for lytic reactivation. Understanding the functional significance of these LANA-interacting host factors in regulating KSHV infection is critical for identifying therapeutic targets and developing potential treatment strategies.
]]></description>
<dc:creator>Castro, M. d. C. C.</dc:creator>
<dc:creator>Adavikolanu, R.</dc:creator>
<dc:creator>Haas, K. M.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Kaake, R. M.</dc:creator>
<dc:creator>Sanchez, E. L.</dc:creator>
<dc:date>2026-01-28</dc:date>
<dc:identifier>doi:10.64898/2026.01.28.699085</dc:identifier>
<dc:title><![CDATA[LANA-dependent Interaction of Host Factors DAXX and BRD4 Impact KSHV Lytic Replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.27.700738v1?rss=1">
<title>
<![CDATA[
SUMO2 Deletion Changes Chromatin Accessibility and Enhances Cytotoxic T Cell Activation and Tumor Infiltration 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.27.700738v1?rss=1</link>
<description><![CDATA[
Cytotoxic T cells (CTL) are crucial for adaptive immunity that leads to prolonged survival and potential cures for cancer. Recent clinical data has shown that pharmacological inhibition of SUMOylation (SUMOi) profoundly modifies tumor microenvironment (TME) and activates CTL, although the mechanism is not well described. In this study, we found that T cell specific knock out (KO) of the most dominant SUMO paralog, Sumo2/SUMO2, in both mouse and human CD8+ T cells significantly enhanced CD8+ T cell activation that is independent of the known mechanism - inducing type I IFN (IFN-I) expression by myeloid cells. Sumo2/SUMO2 KO in CD8+ T cells increased chromatin accessibility for transcription factors BATF, JunB, ATF3, FRA1, FRA2, and AP1 that are known to promote T cell activation and proliferation. Using antigen-specific T cell models, OT1 and Chimeric Antigen Receptor (CAR)-T cells, we found that Sumo2 KO CD8+ T cells had significantly higher tumor infiltration as revealed by flow cytometry, immuno-fluorescence (IF) staining, and single nuclei RNA-sequencing (snRNA-seq) and conferred greater tumor growth inhibition than wildtype (WT) control T cells. snRNA-seq also revealed Sumo2 KO CD8+ T cells increased the expression of Tumor Necrosis Factor-Related Apoptosis-inducing Ligand (TRAIL), induced apoptosis genes in tumor cells and activated IFN-I and IFN-{gamma} responsive genes in all cell types in the TME. These findings elucidate a novel mechanism regarding how SUMOylation can directly control CTL activation and tumor infiltration that activate anti-tumor immunity in the TME. SUMO2 KO can also be a potential strategy to enhance adoptive T cell therapies of solid tumors by enhancing their activity, tumor infiltration and their ability to after the TME.
]]></description>
<dc:creator>Don, N. T. M.</dc:creator>
<dc:creator>Chen, D.-H.</dc:creator>
<dc:creator>Amaral, M. L. J.</dc:creator>
<dc:creator>Ren, T.</dc:creator>
<dc:creator>Kuang, Y.</dc:creator>
<dc:creator>Huang, S.-T.</dc:creator>
<dc:creator>Chilakapati, N.</dc:creator>
<dc:creator>Ren, B.</dc:creator>
<dc:creator>Schoenberger, S. P.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:date>2026-01-29</dc:date>
<dc:identifier>doi:10.64898/2026.01.27.700738</dc:identifier>
<dc:title><![CDATA[SUMO2 Deletion Changes Chromatin Accessibility and Enhances Cytotoxic T Cell Activation and Tumor Infiltration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.28.702361v1?rss=1">
<title>
<![CDATA[
LSD Reconfigures Cortical Dynamics Through Faster Brain Rhythms and Increased Fractal Dimension 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.28.702361v1?rss=1</link>
<description><![CDATA[
Lysergic acid diethylamide (LSD) profoundly alters conscious experience, yet the electrophysiological mechanisms by which it reshapes neural dynamics remain incompletely understood. A hallmark of psychedelic states is widespread cortical desynchronization, typically inferred from reductions in spectral power, but whether such effects reflect genuine weakening of neural oscillations or are confounded by shifts in oscillatory peak frequencies remains unresolved. Here, we address this gap by combining source-resolved magnetoencephalography (MEG), spectral parameterization, temporal complexity metrics, and interpretable machine learning in an LSD versus placebo design, with and without music. We show that LSD induces robust, spatially structured increases in alpha and beta peak frequencies alongside genuine attenuation of oscillatory power, with these effects displaying partly dissociable cortical patterns. Beyond rhythmic activity, LSD is associated with flattening of the aperiodic 1/f spectral slope and increased neural signal fractality and complexity, preferentially affecting sensory, language, emotion, and imagery-related networks while sparing motor cortex. Machine-learning analyses further identify peak-frequency shifts, aperiodic parameters, and complexity measures as key discriminators of the psychedelic state. Music does not robustly amplify these neural signatures and instead shows a trend toward attenuation. Together, these findings provide a comprehensive electrophysiological account of how LSD reorganizes large-scale human brain dynamics and highlight features that may differentiate its neural signature from that of other psychedelics.
]]></description>
<dc:creator>Subramani, V.</dc:creator>
<dc:creator>Nest, T.</dc:creator>
<dc:creator>Pascarella, A.</dc:creator>
<dc:creator>Brunel, J.</dc:creator>
<dc:creator>Ramos, Y. J. M.</dc:creator>
<dc:creator>Harel, Y.</dc:creator>
<dc:creator>Muthukumaraswamy, S. D.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Lioi, G.</dc:creator>
<dc:creator>Farrugia, N.</dc:creator>
<dc:creator>Jerbi, K.</dc:creator>
<dc:date>2026-01-29</dc:date>
<dc:identifier>doi:10.64898/2026.01.28.702361</dc:identifier>
<dc:title><![CDATA[LSD Reconfigures Cortical Dynamics Through Faster Brain Rhythms and Increased Fractal Dimension]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.28.702385v1?rss=1">
<title>
<![CDATA[
A Single-Cell and Spatial 3D Multi-omic Atlas of Developing Human Basal Ganglia and Inhibitory Neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.28.702385v1?rss=1</link>
<description><![CDATA[
The human basal ganglia (BG), subcortical nuclei fundamental to motor regulation and cognitive modulation, is constructed from neurons produced during gestation in the adjacent ganglionic eminences (GEs). GEs are transient structures in the ventral prenatal brain that also generate GABAergic inhibitory neurons which migrate to destinations in the BG, cortex and other destinations. This study aims to elucidate the epigenomic and 3D-genomic dynamics involved in the specification and maturation of GEs and GE-derived neurons, using single-nucleus methyl-3C sequencing (snm3C-seq), highly-multiplexed spatial transcriptomics, and chromatin+RNA single-molecule imaging. Our multi-modal data support a heterogeneous temporal progression across GE subregions, with the lateral GE (LGE) showing declining neurogenic activity in mid-gestation and caudal GE (CGE) exhibiting ongoing developmental progression through infancy. We identified regulatory programs that specify subtypes of BG principal cells, medium spiny neurons (MSN), via synchronized maturation of the 3D-epigenome. In infant brains, we found a transient short-range enriched (SE) chromatin conformation during the transition between oligodendrocyte progenitors (OPCs) and oligodendrocytes (ODCs), and a temporary shift toward Long-range Enriched (LE) chromatin conformation in projection neurons, extending previous works showing the differentiation of neurons and glial cells is associated with permanent SE and LE conformation, respectively. Lastly, we found that gene regulatory regions active in MSNs were enriched in loci associated with genetic risk for neuropsychiatric disease. Our study delineates the highly complex, lineage-specific 3D genomic dynamics in ventral progenitors and basal ganglia populations of the perinatal human brain.

HighlightsO_LIJoint 3D genome and DNA methylome analysis of ventral brain progenitor zones
C_LIO_LIHeterogeneous developmental progressions of the ganglionic eminences
C_LIO_LIDistinct development dynamics and regulatory landscape of MSNs and interneurons
C_LIO_LITransient remodeling of the 3D-genome in neurons and oligodendrocyte progenitors
C_LI
]]></description>
<dc:creator>Heffel, M. G.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Pastor-Alonso, O.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Baig, M. S.</dc:creator>
<dc:creator>Irfan Ghoor, R.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Kern, C.</dc:creator>
<dc:creator>Kum, J.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Paino, J.</dc:creator>
<dc:creator>Tsai, M. J.</dc:creator>
<dc:creator>Tai, C.-Y.</dc:creator>
<dc:creator>Tucker, G.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Hou, A.</dc:creator>
<dc:creator>von Behren, Z.</dc:creator>
<dc:creator>Bhade, M.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Sandoval, K.</dc:creator>
<dc:creator>Scholes, J.</dc:creator>
<dc:creator>Codrea, F.</dc:creator>
<dc:creator>Calimlim, J.</dc:creator>
<dc:creator>Liao, E. K.</dc:creator>
<dc:creator>Leung, G.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Eskin, E.</dc:creator>
<dc:creator>Flint, J.</dc:creator>
<dc:creator>Cotter, J. A.</dc:creator>
<dc:creator>Pasaniuc, B.</dc:creator>
<dc:creator>Bintu, B.</dc:creator>
<dc:creator>Zhu, Q.</dc:creator>
<dc:creator>Mukamel, E. A.</dc:creator>
<dc:creator>Ernst, J.</dc:creator>
<dc:creator>Paredes, M. F.</dc:creator>
<dc:creator>Luo, C.</dc:creator>
<dc:date>2026-01-29</dc:date>
<dc:identifier>doi:10.64898/2026.01.28.702385</dc:identifier>
<dc:title><![CDATA[A Single-Cell and Spatial 3D Multi-omic Atlas of Developing Human Basal Ganglia and Inhibitory Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.27.702131v1?rss=1">
<title>
<![CDATA[
A genetically encoded ionic-stress sensor reveals protons as a sleep driver 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.27.702131v1?rss=1</link>
<description><![CDATA[
Dynamic ionic changes are hallmarks of physiological and behavioral state transitions, including sleep in animals. Although biosensors for specific cellular ions are widely available, real-time monitoring of overall ionic strength in living organisms remains challenging. Here, we present a genetically encoded nuclear translocation ionic sensor (GENTIS) that enables direct visualization of ionic stress in vivo. Using GENTIS in C. elegans, we uncover rhythmic elevations in ionic strength during larval molting transitions that coincide with the lethargus sleep. Cytosolic proton ionic increase through inhibition of v-ATPase is sufficient to induce GENTIS nuclear translocation and evoke behavioral quiescence, characterized by reduced feeding and activation of sleep-active neurons. Apical membrane v-ATPases undergo disassembly during lethargus and under sleep-inducing stress conditions, leading to proton accumulation. Notably, this proton-mediated sleep is suppressed by proton buffering with ammonium. Together, these findings establish GENTIS as a powerful tool for tracking ionic strength dynamics in living organisms and support protons as a physiological driver of sleep.
]]></description>
<dc:creator>Ji, Z.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Wei, S.</dc:creator>
<dc:creator>Bian, Y.</dc:creator>
<dc:creator>Zeng, W.</dc:creator>
<dc:creator>Chung, C.-I.</dc:creator>
<dc:creator>Ma, Z.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Shu, X.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2026-01-30</dc:date>
<dc:identifier>doi:10.64898/2026.01.27.702131</dc:identifier>
<dc:title><![CDATA[A genetically encoded ionic-stress sensor reveals protons as a sleep driver]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.28.702329v1?rss=1">
<title>
<![CDATA[
Methionine metabolism and the NOP2 methyltransferase are essential for MYC-Driven liver tumorigenesis. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.28.702329v1?rss=1</link>
<description><![CDATA[
Hepatocellular carcinoma (HCC) represents the third leading cause of cancer-related death worldwide and has been increasing in developed nations.1,2 The MYC oncogene or its paralogs are frequently amplified or overexpressed in subtypes of cancer associated with stem cell-like features and worse clinical outcomes,3,4 including in liver cancer.5 Unfortunately, selective inhibitors that target MYC or its transcriptional program are not yet clinically available for therapy of HCC. Here, we identified methionine metabolism as a selective vulnerability for MYC but not RAS-driven liver cancers. MYC-driven liver cancer cells are methionine dependent, with markedly diminished tumor growth when mice are fed a methionine low diet. While RAS-driven liver cancer was resistant to a low methionine diet. S-adenosylmethionine (SAM), the predominant methyl donor, partially rescues cell proliferation following methionine depletion, suggesting that methylation processes are especially critical in the context of MYC high tumor cells. Heavy isotope methionine tracing in MYC high cells identified increased levels of m5C nucleotides. We found NOP2, an rRNA m5C-methyltransferase, was regulated by both MYC overexpression and methionine abundance linking the two processes. Methionine depletion reduced methylation of multiple 28S rRNA residues as did NOP2 knockdown. Depletion of NOP2 selectively inhibited MYC liver cancer cell proliferation and in vivo tumor growth. Thus, methionine catabolism is critical for MYC-driven liver tumorigenesis and the rRNA methyltransferase NOP2 may serve as a new therapeutic target in liver cancer.
]]></description>
<dc:creator>Lin, S.</dc:creator>
<dc:creator>Berdan, C.</dc:creator>
<dc:creator>Sandy, M.</dc:creator>
<dc:creator>Lu, X.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Nomura, D.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Lee, J. V.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:date>2026-01-30</dc:date>
<dc:identifier>doi:10.64898/2026.01.28.702329</dc:identifier>
<dc:title><![CDATA[Methionine metabolism and the NOP2 methyltransferase are essential for MYC-Driven liver tumorigenesis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.29.702647v1?rss=1">
<title>
<![CDATA[
H4K20me3 and CTCF act reciprocally at TAD boundaries to regulate cell state transitions 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.29.702647v1?rss=1</link>
<description><![CDATA[
Chromatin is organized into topologically associating domains (TADs) that are critical for gene regulation. Transitions between cell states, such as proliferation and quiescence, involve genome-scale chromatin dynamics and transcriptional changes, but the underlying mechanisms are not well-understood. Here we show that the reversible shift between proliferation and quiescence is regulated by the opposing action of CTCF and the histone modification H4K20me3. CTCF is a boundary element that defines TADs, while H4K20me3 has previously been associated with heterochromatin, chromatin compaction, and repetitive elements. Using CUT&Tag, RNA-seq, and functional perturbations, we demonstrate that H4K20me3 and CTCF compete for chromatin, including at TAD boundary elements, with increased H4K20me3 in quiescent cells antagonizing CTCF activity in proliferating cells. Manipulation of H4K20me3 levels shows that in quiescent cells, elevated H4K20me3 alters the chromatin landscape, leading to more compact chromatin architecture, elongated nuclei, and induction of a quiescence gene expression program. Conversely, CTCF binding is associated with open chromatin conformation, rounder nuclei, expression of proliferation-associated genes, lower levels of quiescence marker p27/CDKN1B, and cell division in cultured cells and mice. Fibroblasts deficient for H4K20me3 methyltransferase KMT5C are more proliferative, with reduced expression of quiescence genes, and KMT5C-deficient mice are larger. Our findings reveal a reversible antagonistic interplay between H4K20me3 and CTCF that modulates the functional outcome of chromatin architecture to effect changes in cellular state. This new paradigm for regulation of the proliferation-quiescence transition suggests a molecular basis for common developmental transitions and disorders of proliferation.

HighlightsO_LIH4K20me3, a mark associated with heterochromatin, and CTCF compete for chromatin boundary elements
C_LIO_LICTCF and H4K20me3 play opposing roles, with increased CTCF in proliferating cells and elevated H4K20me3 in quiescent cells
C_LIO_LIModulating H4K20me3 or CTCF reversibly shifts cells between proliferative and quiescent state
C_LIO_LIMice with reduced H4K20me3 are larger, consistent with hyperproliferation
C_LI
]]></description>
<dc:creator>Atai, K.</dc:creator>
<dc:creator>Mitra, M.</dc:creator>
<dc:creator>Bohn, E.</dc:creator>
<dc:creator>Hu, R.</dc:creator>
<dc:creator>Corvalan, A.</dc:creator>
<dc:creator>Jelinek, D.</dc:creator>
<dc:creator>Voisin, L.</dc:creator>
<dc:creator>Ambrus, A.</dc:creator>
<dc:creator>Sarathy, K.</dc:creator>
<dc:creator>Sparks, W. M.</dc:creator>
<dc:creator>Bonitto, K.</dc:creator>
<dc:creator>Dunkwu, A.</dc:creator>
<dc:creator>Coller, H. A.</dc:creator>
<dc:date>2026-01-30</dc:date>
<dc:identifier>doi:10.64898/2026.01.29.702647</dc:identifier>
<dc:title><![CDATA[H4K20me3 and CTCF act reciprocally at TAD boundaries to regulate cell state transitions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.29.702705v1?rss=1">
<title>
<![CDATA[
Predicting early bactericidal activity of tuberculosis drug combinations using a translational pharmacokinetic-pharmacodynamic modeling approach 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.29.702705v1?rss=1</link>
<description><![CDATA[
Phase IIa pulmonary tuberculosis (TB) trials typically assess the early bactericidal activity (EBA) of monotherapy for over 14 days. However, few studies have evaluated drug combinations, even though optimal monotherapy doses may not directly translate to combinations. Translational pharmacokinetic-pharmacodynamic (PK-PD) modeling has shown promise in predicting human treatment responses based on preclinical monotherapy data; however, its application in drug combinations remains limited. This study aimed to extend and validate our previously developed translational monotherapy PK-PD modeling platform to predict the EBA of two-drug combinations. Interactions between bedaquiline, pretomanid, linezolid, and pyrazinamide were characterized using two modeling approaches: the empirical SUPER method and the mechanistic General Pharmacodynamic Interaction model. Both approaches were independently linked to our translational platform and validated using mouse data and Phase IIa clinical results from the NC-001 study. Both modeling methods identified consistent interaction patterns, including antagonistic interactions when bedaquiline was combined with either pretomanid or linezolid. Pyrazinamide has emerged as the most effective companion for both bedaquiline and pretomanid. Our platform reasonably predicted 14-day clinical sputum colony-forming unit counts for multiple two-drug combinations, with most observations falling within the 95% prediction intervals, supporting its use in accelerating regimen development. Our study demonstrated that the translational PK-PD platform reliably predicts both short- and long-term outcomes for combinations, regardless of the interaction model. This supports its application across drug development stages to inform dose selection and effective companion drugs for anti-TB therapies.
]]></description>
<dc:creator>de Castro Suarez, N.</dc:creator>
<dc:creator>Nuermberger, E. L.</dc:creator>
<dc:creator>Ernest, J.</dc:creator>
<dc:creator>Savic, R. M.</dc:creator>
<dc:date>2026-02-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.29.702705</dc:identifier>
<dc:title><![CDATA[Predicting early bactericidal activity of tuberculosis drug combinations using a translational pharmacokinetic-pharmacodynamic modeling approach]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.30.702717v1?rss=1">
<title>
<![CDATA[
Hydrogel-imposed boundary conditions guide single-lumen neuroepithelial morphogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.30.702717v1?rss=1</link>
<description><![CDATA[
Three-dimensional (3D) stem cell-based cultures have emerged as promising in vitro model systems for studying human neurodevelopment. Current neural organoid protocols lack well-defined extracellular matrix (ECM) signaling and are limited by the formation of irregular tissue morphologies with multiple organizing centers, in contrast to the single neuroepithelial structure that emerges during embryonic development. This variability limits inter-organoid reproducibility and constrains their utility for modeling early developmental processes. To overcome these limitations, we leverage a materials-based approach to impose dynamic boundary conditions that extrinsically guide the self-organization of human induced pluripotent stem cells (iPSCs). Specifically, we develop a family of hyaluronic acid-elastin-like protein (HELP) hydrogels crosslinked with dynamic covalent bonds that recapitulate key biochemical and biophysical properties of the developing human neural ECM. Within these HELP hydrogels, iPSCs robustly self-organize from a single cell into complex neuroepithelial tissues with a single lumen. By tuning the boundary conditions imposed by the hydrogel, we identify matrix stress relaxation rate and tensional homeostasis as key regulators of single-lumen rosette formation and maintenance. With this hydrogel-enabled system, we identify phenotypic abnormalities in an early neurodevelopmental model of 22q11.2 deletion syndrome. Ultimately, our tunable engineered hydrogel supports the initiation of single-cell derived 3D neuroepithelial tissues, enables investigation into how matrix-imposed boundary conditions guide developmental morphogenesis, and establishes a reproducible platform for disease modeling.
]]></description>
<dc:creator>Huang, M. S.</dc:creator>
<dc:creator>Roth, J. G.</dc:creator>
<dc:creator>Kim, D.</dc:creator>
<dc:creator>Pashin, K. P.</dc:creator>
<dc:creator>Pizzarella, D.</dc:creator>
<dc:creator>Yang, T. M.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Navarro, R. S.</dc:creator>
<dc:creator>Palmer, T. D.</dc:creator>
<dc:creator>Heilshorn, S. C.</dc:creator>
<dc:date>2026-02-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.30.702717</dc:identifier>
<dc:title><![CDATA[Hydrogel-imposed boundary conditions guide single-lumen neuroepithelial morphogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.30.702860v1?rss=1">
<title>
<![CDATA[
cGAS-mediated IFN-I signaling contributes to disease progression in drug-refractory epilepsy 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.30.702860v1?rss=1</link>
<description><![CDATA[
Epilepsy is a prevalent neurological disease with a third of patients becoming non-responsive to antiepileptic drugs and developing drug-refractory epilepsy (DRE). Here we report that DRE disease progression is contributed by overactive cyclic GMP-AMP synthase (cGAS), a double-stranded DNA sensor that induces type I interferon (IFN-I) signaling. In human DRE microglia, we observe a robust IFN-I signature and the activation of upstream cGAS-STING signaling. Further, in mouse models of Dravet syndrome, a genetic form of DRE, we observe the activation of the cGAS pathway. We show that microglial cGAS can be activated by DNA released from hyperexcitable neurons. Genetic reduction and pharmacological inhibition of cGAS reduces epileptic phenotypes, glial inflammatory signatures, and neuronal transcriptomic changes, underscoring the therapeutic potential of targeting cGAS for DRE treatment.
]]></description>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Fan, L.</dc:creator>
<dc:creator>Wong, M. Y.</dc:creator>
<dc:creator>Lei, Z.</dc:creator>
<dc:creator>Krishnamachary, B.</dc:creator>
<dc:creator>Zhu, D.</dc:creator>
<dc:creator>Cadiz, M.</dc:creator>
<dc:creator>Nagiri, R. K.</dc:creator>
<dc:creator>Ye, P.</dc:creator>
<dc:creator>Norman, K.</dc:creator>
<dc:creator>Bhagwat, M.</dc:creator>
<dc:creator>Lee, Y. J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Amin, S.</dc:creator>
<dc:creator>Lauderdale, K.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Luo, W.</dc:creator>
<dc:creator>Gong, S.</dc:creator>
<dc:creator>Liechty, B. L.</dc:creator>
<dc:creator>Palop, J. J.</dc:creator>
<dc:creator>Sinha, S. C.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Gan, L.</dc:creator>
<dc:date>2026-02-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.30.702860</dc:identifier>
<dc:title><![CDATA[cGAS-mediated IFN-I signaling contributes to disease progression in drug-refractory epilepsy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.01.702869v1?rss=1">
<title>
<![CDATA[
Cellular diversity of the developing chick trigeminal ganglion at single-cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.01.702869v1?rss=1</link>
<description><![CDATA[
BackgroundThe trigeminal ganglion (TG) is a structure of the peripheral nervous system, composed of neuronal and non-neuronal cell types, that integrates sensory input from the face and jaw. The developing TG is derived from two embryonic cell populations: neural crest and cranial placode. Both populations play critical roles in TG development and must interact to coordinate changes in gene expression that regulate specification, differentiation, and organization. However, the molecular characteristics of the heterogeneous cell populations within the developing TG remain poorly defined.

ResultsWe performed single-cell RNA-sequencing (scRNA-seq) on TG from developing chick embryos at HH17. Our high-resolution dataset (14 clusters, [~]87000 cells) provides insight into cellular diversity within the developing TG. As expected, we identified placode-derived neurons as well as neural crest cells prior to neuronal differentiation. In addition to classic markers, we identified novel transcripts with unknown roles in TG development, including several long non-coding RNAs (lncRNAs).

ConclusionsWe generated a single-cell atlas of the developing chick trigeminal ganglion during early axonogenesis and defined the transcriptomic states of its diverse cell populations. Our results provide a useful resource for better understanding the cell populations contributing to TG development and gene expression that drives cell identity and differentiation.
]]></description>
<dc:creator>Arul Nambi Rajan, A.</dc:creator>
<dc:creator>Hutchins, E. J.</dc:creator>
<dc:date>2026-02-03</dc:date>
<dc:identifier>doi:10.64898/2026.02.01.702869</dc:identifier>
<dc:title><![CDATA[Cellular diversity of the developing chick trigeminal ganglion at single-cell resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.01.703132v1?rss=1">
<title>
<![CDATA[
Structure-guided design of a targeted autoantibody degrader for neurologic disease 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.01.703132v1?rss=1</link>
<description><![CDATA[
Despite rapid progress in the diagnosis of autoantibody-mediated neurologic diseases, standard-of-care therapeutic options remain limited to nonspecific immunosuppression. Here, we report an alternative therapeutic strategy using targeted protein degradation to eliminate pathogenic autoantibodies while leaving the rest of the immune system intact. We previously discovered autoimmune vitamin B12 central deficiency (ABCD), a neurologic condition in which autoantibodies targeting the transcobalamin receptor (CD320) impair the transport of cobalamin (B12) from the blood into the central nervous system (CNS). Combining scanning alanine mutagenesis by phage display, cryo-electron microscopy, and computational modeling, we elucidated a highly conserved anti-CD320 epitope and defined the structural determinants of antigen-autoantibody binding. Next, we synthesized a lysosome-targeting chimera (LYTAC) comprising the lysosome targeting glycan, triGalNAc, fused to the antigenic epitope of CD320 as autoantibody bait. In vitro, this LYTAC promoted the specific lysosomal internalization and extracellular clearance of anti-CD320, restoring homeostatic cellular uptake of B12. In a passive transfer mouse model of ABCD, LYTAC treatment rapidly cleared anti-CD320 from circulation and prevented penetration of anti-CD320 into the CNS. These findings uncover the mechanism of autoantibody-antigen binding in ABCD and demonstrate targeted autoantibody degradation as a therapeutic strategy that may be generalizable to other autoimmune neurologic diseases.
]]></description>
<dc:creator>Zimanyi, M.</dc:creator>
<dc:creator>Dayao, M.</dc:creator>
<dc:creator>Asencor, A. I.</dc:creator>
<dc:creator>Kondapavulur, S.</dc:creator>
<dc:creator>Asaki, J.</dc:creator>
<dc:creator>McCutcheon, K.</dc:creator>
<dc:creator>Dabaco, C.</dc:creator>
<dc:creator>Bodansky, A.</dc:creator>
<dc:creator>Craik, C.</dc:creator>
<dc:creator>Pleasure, S.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:creator>Cheng, Y.</dc:creator>
<dc:creator>Wilson, M.</dc:creator>
<dc:creator>Pluvinage, J. V.</dc:creator>
<dc:date>2026-02-03</dc:date>
<dc:identifier>doi:10.64898/2026.02.01.703132</dc:identifier>
<dc:title><![CDATA[Structure-guided design of a targeted autoantibody degrader for neurologic disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.02.700737v1?rss=1">
<title>
<![CDATA[
LUCas: Light-Uncaged Cas13a using photocleavable interfering guide RNAs 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.02.700737v1?rss=1</link>
<description><![CDATA[
CRISPR diagnostics have emerged as powerful tools for detecting infectious diseases, with the RNA endonuclease Cas13a enabling sensitive and specific, amplification-free RNA detection through collateral trans-cleavage of fluorescent reporters. However, background cleavage from unbound enzyme, contaminating nucleases, and unsynchronized initiation of reactions limits assay sensitivity and interpretability. A strategy to precisely control the onset of Cas13a catalytic activity, essentially a molecular "starting gun", would address these challenges and expand assay design space. Here, we introduce Light-Uncaged Cas13a (LUCas), a light controllable system that directly gates Cas13a using a photocleavable interfering guide RNA (pc-igRNA) that suppresses trans-cleavage activity even in the presence of target RNA. Brief UV illumination releases this suppression, restoring full activity. Quantitative kinetic analysis reveals an approximately 100-fold suppression of trans-cleavage activity prior to photo-activation. Importantly, LUCas also suppresses target-independent background activity, enabling a predictive, background-limited determination of assay sensitivity. Using measured kinetic parameters, we predict and experimentally validate the limit-of-detection of the LUCas system. Finally, we demonstrate a multiplexed detection strategy termed "temporal barcoding," which enables quantitative detection of viral co-infections in a single bulk reaction. Together, these results establish LUCas as a general framework for mechanistically informed, light-based control of Cas13a activity.
]]></description>
<dc:creator>Ng, C. F.</dc:creator>
<dc:creator>Krishnamurthy, D.</dc:creator>
<dc:creator>Dextre, A.</dc:creator>
<dc:creator>Chorlay, A.</dc:creator>
<dc:creator>Ott, M.</dc:creator>
<dc:creator>Fletcher, D. A.</dc:creator>
<dc:date>2026-02-04</dc:date>
<dc:identifier>doi:10.64898/2026.02.02.700737</dc:identifier>
<dc:title><![CDATA[LUCas: Light-Uncaged Cas13a using photocleavable interfering guide RNAs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.02.703317v1?rss=1">
<title>
<![CDATA[
The selectivity implications of docking libraries with greater and lesser similarities to bio-like molecules 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.02.703317v1?rss=1</link>
<description><![CDATA[
As virtual libraries have expanded into the tens of billions via make-on-demand chemistry, their similarity to metabolites, natural products, and drugs ("bio-like" molecules) has rapidly diminished. Despite this divergence, molecular docking of these ultra-large libraries has yielded molecules at higher experimental hit-rates and with improved affinities. The structural divergence from bio-like space raises the possibility that molecules from these ultra-large libraries have improved selectivity. Just as plausibly, if hit-rates on-target are divorced from similarity to bio-like molecules, so too may be selectivity against off-targets. Here, we test whether docking hits for the 5-HT2A serotonin receptor from ultra-large libraries are more selective than those from smaller and more bio-like "in-stock" libraries. Chemoinformatic similarity predicts that docking actives from the in-stock library have more off-targets than the more chemically novel hits emerging from docking the ultra-large library. This may reflect the bias of the known, however, as when tested experimentally at scale against 318 GPCRs, both 16 agonists from the ultra-large library and 20 actives from the in-stock library had similar numbers of off-targets. While the ultra-large library hits are more sub-type selective for the 5-HT2A over the 5-HT2B and 5-HT2C receptors, overall these results may suggest that selectivity against off-targets, like affinity and hit-rates for on-targets, is divorced from library similarity to bio-like molecules.
]]></description>
<dc:creator>Hall, B. W.</dc:creator>
<dc:creator>Sakamoto, K.</dc:creator>
<dc:creator>Huang, X.-P.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Roth, B. L.</dc:creator>
<dc:date>2026-02-04</dc:date>
<dc:identifier>doi:10.64898/2026.02.02.703317</dc:identifier>
<dc:title><![CDATA[The selectivity implications of docking libraries with greater and lesser similarities to bio-like molecules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.02.703338v1?rss=1">
<title>
<![CDATA[
NLRP11 promotes non-canonical inflammasome activation in human macrophages by enhancing caspase-4 recognition of cytosolic lipopolysaccharide 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.02.703338v1?rss=1</link>
<description><![CDATA[
Innate immune detection of Gram-negative bacteria depends on sensing of cytosolic lipopolysaccharide (cLPS) by the non-canonical inflammasome, mediated in humans by NLRP11 and caspase-4 (CASP4). Activation of this pathway in human macrophages triggers gasdermin-D activation and pyroptotic cell death. Although CASP4 directly binds LPS in vitro, additional host factors are required for efficient activation in vivo. Here, we show that NLRP11, a primate-specific pattern recognition receptor, facilitates CASP4 recognition of cLPS and promotes non-canonical inflammasome activation. NLRP11 functions upstream CASP4, forming an ASC-independent complex that requires a conserved CASP4 p20 residue, binds cLPS, and enhances CASP4-dependent LPS recognition. Mutational analyses demonstrate that in human macrophages, in addition to LPS binding and CASP4 catalytic activity, CASP4 interaction with NLRP11 is essential for efficient pyroptosis. Together, these findings establish NLRP11 as a primate-specific determinant that enhances CASP4-mediated cLPS detection and non-canonical inflammasome activation, revealing a mechanism for human-specific regulation of innate immunity.
]]></description>
<dc:creator>Gil-Marques, M. L.</dc:creator>
<dc:creator>Devant, P.</dc:creator>
<dc:creator>Kagan, J. C.</dc:creator>
<dc:creator>Goldberg, M. B.</dc:creator>
<dc:date>2026-02-04</dc:date>
<dc:identifier>doi:10.64898/2026.02.02.703338</dc:identifier>
<dc:title><![CDATA[NLRP11 promotes non-canonical inflammasome activation in human macrophages by enhancing caspase-4 recognition of cytosolic lipopolysaccharide]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.03.703379v1?rss=1">
<title>
<![CDATA[
Platelet Molecular Maturation Links Platelet Aging and the Platelet Storage Lesion 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.03.703379v1?rss=1</link>
<description><![CDATA[
Human platelets change over their 7-10 day lifespan, yet the molecular mechanisms underlying platelet aging remain poorly defined. Using two independent RNA sequencing datasets of fluorescence-activated cell sorted young and old human platelets, we developed a unified transcriptomic model to characterize RNA metabolism across the platelet lifespan, which we termed platelet molecular maturation. This was applied to RNA sequencing data from room-temperature stored platelets (up to 7 days) and cold-stored platelets (7, 14, or 21 days). We identified highly concordant aging signatures, including 6,015 shared expressed genes and 2,008 shared differentially expressed genes (DEGs) with strongly correlated fold changes, demonstrating a conserved platelet aging program. Nucleotide-level analyses revealed preferential 3'-directed degradation among downregulated transcripts during endogenous platelet aging and room-temperature storage, supporting an organized RNA decay process that was correlated with platelet function changes. Room-temperature storage recapitulated platelet molecular maturation, showing concordance with aging-related gene expression changes and enrichment of downregulated gene sets. In contrast, cold-storage significantly attenuated platelet molecular maturation and 3'-directed degradation. A total of 669 genes were consistently differentially expressed between room-temperature and cold-stored platelets, while no DEGs were detected during cold-storage, indicating transcriptional stability. Platelet transcript stability in cold-storage correlated with preserved platelet hemostatic function. These findings establish platelet molecular maturation as a conserved, functionally relevant model linking endogenous platelet aging to platelet storage lesions and providing mechanistic insight into preserved platelet hemostatic function in cold-storage. This atlas of platelet RNA metabolism supports biomarker discovery and strategies to improve storage.

KEY POINTSO_LIBy integrating multiple high-quality RNA sequencing datasets with novel analytic approaches tailored to the biology of anucleate platelets, we show that platelet aging is not a passive process of transcript decay, but follows a structured and reproducible molecular trajectory both endogenously and in storage, which we term platelet molecular maturation.
C_LIO_LIStorage temperature emerged as a dominant modifier of this trajectory, with cold-storage markedly slowing RNA metabolic kinetics and preserving transcripts associated with younger, more hemostatically competent platelets.
C_LIO_LITogether, these findings provide mechanistic insight into the platelet storage lesion and identify transcriptomic features that may serve as biomarkers or therapeutic targets to extend platelet shelf life.
C_LI
]]></description>
<dc:creator>Bainton, C. M.</dc:creator>
<dc:creator>Santos, Y.</dc:creator>
<dc:creator>Mayer, F.</dc:creator>
<dc:creator>Fields, A.</dc:creator>
<dc:creator>Zoghbi, K.</dc:creator>
<dc:creator>Mayer, N.</dc:creator>
<dc:creator>Williamson, D.</dc:creator>
<dc:creator>Chinn, G.</dc:creator>
<dc:creator>Rahn, K. C.</dc:creator>
<dc:creator>Musgrove, J. M.</dc:creator>
<dc:creator>Thomas, K. A.</dc:creator>
<dc:creator>Sall, J.</dc:creator>
<dc:creator>Shea, S. M.</dc:creator>
<dc:creator>Kornblith, L. Z.</dc:creator>
<dc:creator>Bainton, R. J.</dc:creator>
<dc:date>2026-02-05</dc:date>
<dc:identifier>doi:10.64898/2026.02.03.703379</dc:identifier>
<dc:title><![CDATA[Platelet Molecular Maturation Links Platelet Aging and the Platelet Storage Lesion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.04.703058v1?rss=1">
<title>
<![CDATA[
Scalable probe-based single-cell transcriptional profiling for virtual cell perturbation mapping and synthetic biology phenotyping 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.04.703058v1?rss=1</link>
<description><![CDATA[
Large-scale single-cell transcriptional phenotyping of genetic perturbations (perturb-seq) links genes to phenotypes and should enable virtual cell predictive modeling and cellular engineering. However, current perturb-seq single-cell methods are costly, information sparse and require barcodes for many applications. We developed ProPer-seq, a perturb-seq method that uses multiplexed custom DNA probe panels to measure and phenotype synthetic biology perturbations at single-cell resolution without barcodes, including multidomain proteins and sgRNAs. ProPer-seq faithfully reproduces gold-standard perturb-seq phenotypes while achieving 4-fold cost reduction and 50% increased gene detection per cell. As a scalable fixed-cell profiling method, ProPer-seq enables atlas-scale profiling for virtual-cell initiatives and demonstrates data quality suitable for training and validating predictive models. Lastly, ProPer-seqs targeted detection of modular transgenes enables library-on-library perturbation profiling of combinatorial synthetic protein design spaces. We applied this to 3,550 sgRNA x dCas9 effector combinations as well as 260 CAR x ORF combinations dynamically profiled in primary T cells, revealing principles of transcriptional control and cell state modulation by multidomain synthetic transgenes.
]]></description>
<dc:creator>Swinderman, J. T.</dc:creator>
<dc:creator>Tung, P.-Y.</dc:creator>
<dc:creator>Winters, A.</dc:creator>
<dc:creator>Goudy, L.</dc:creator>
<dc:creator>Wilson, C. M.</dc:creator>
<dc:creator>Bounds, L. R.</dc:creator>
<dc:creator>Teyssier, N.</dc:creator>
<dc:creator>Agrawal, A.</dc:creator>
<dc:creator>Dobin, A.</dc:creator>
<dc:creator>Hua, T.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Feng, F. Y.</dc:creator>
<dc:creator>Marson, A.</dc:creator>
<dc:creator>Burke, D. P.</dc:creator>
<dc:creator>Hsu, P. D.</dc:creator>
<dc:creator>Roohani, Y. H.</dc:creator>
<dc:creator>Konermann, S.</dc:creator>
<dc:creator>Kosicki, M.</dc:creator>
<dc:creator>Li, N.</dc:creator>
<dc:creator>Gilbert, L. A.</dc:creator>
<dc:date>2026-02-05</dc:date>
<dc:identifier>doi:10.64898/2026.02.04.703058</dc:identifier>
<dc:title><![CDATA[Scalable probe-based single-cell transcriptional profiling for virtual cell perturbation mapping and synthetic biology phenotyping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.04.700910v1?rss=1">
<title>
<![CDATA[
Stereoselective methyl-swapping demonstrates target specificity of cognitive enhancer 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.04.700910v1?rss=1</link>
<description><![CDATA[
The Integrated Stress Response (ISR) couples cellular stress sensing to translational control, playing a critical role in the homeostatic regulation of cell health. However, prolonged and unmitigated ISR activation becomes maladaptive and drives the progression of a wide range of pathologies, including cognitive decline. Pharmacological inhibition of the ISR with the small, drug-like molecule ISRIB has proven remarkably effective in reversing cognitive deficits and pathology in animal models, highlighting its potential for therapeutic intervention in humans. We engineered an allele-specific ISRIB analog (mISRIB) that selectively targets a mutant form of eIF2B, the molecular target of ISRIB, without affecting wild-type eIF2B. Notably, mISRIB treatment in mice homozygous for the eIF2B mutant allele enhances synaptic plasticity and long-term memory, confirming the on-target mechanism underlying ISRIBs cognitive benefits. Our results provide a framework for dissecting the ISRs contributions within complex cellular networks, such as those governing brain function, with precise temporal and spatial resolution.
]]></description>
<dc:creator>Boone, M.</dc:creator>
<dc:creator>Dalwadi, U.</dc:creator>
<dc:creator>Deal, A.</dc:creator>
<dc:creator>Zhu, P. J.</dc:creator>
<dc:creator>Croll, T. I.</dc:creator>
<dc:creator>Yamazaki, M.</dc:creator>
<dc:creator>Prescott, K.</dc:creator>
<dc:creator>Minopoli, R.</dc:creator>
<dc:creator>Biscocho, I.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Lee, D. J.</dc:creator>
<dc:creator>Arthur, C. P.</dc:creator>
<dc:creator>Laughlin, T. G.</dc:creator>
<dc:creator>Zhou, H.</dc:creator>
<dc:creator>Klope, M. T.</dc:creator>
<dc:creator>Egea, P. F.</dc:creator>
<dc:creator>Schoof, M.</dc:creator>
<dc:creator>Lawrence, R.</dc:creator>
<dc:creator>Renslo, A. R.</dc:creator>
<dc:creator>Costa-Mattioli, M.</dc:creator>
<dc:creator>Frost, A.</dc:creator>
<dc:creator>Walter, P.</dc:creator>
<dc:date>2026-02-07</dc:date>
<dc:identifier>doi:10.64898/2026.02.04.700910</dc:identifier>
<dc:title><![CDATA[Stereoselective methyl-swapping demonstrates target specificity of cognitive enhancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.06.704456v1?rss=1">
<title>
<![CDATA[
A Large Animal Model of Heritable Pulmonary Arterial Hypertension UsingGene-edited BMPR2 Sheep 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.06.704456v1?rss=1</link>
<description><![CDATA[
Pulmonary Arterial Hypertension (PAH) is a rare vascular disorder characterized by elevated pressure in pulmonary arteries, eventually leading to right ventricular failure. Approximately 50% of pediatric disease and 20% of adult disease can be linked to a genetic mutation, with nearly 70% of these cases involving mutations in the bone morphogenetic protein receptor type 2 (BMPR2) locus. Investigations using rodent models have made significant advances in our understanding of BMPR2 signaling; however, limited data exist regarding the onset and course of PAH, and etiologies for phenotypic expression in these patients remain unknown. In this work, we describe the development of a novel ovine model of heritable PAH. Because homozygous disruption of BMPR2 is embryonic lethal, we developed heterozygous BMPR2 sheep by using a PAM-disrupting synonymous single stranded oligodeoxyribonucleotide alongside a single guide RNA and Cas9 mediated gene editing strategy. The resulting BMPR2(+/-) lambs demonstrated cardiac and pulmonary vascular pathology that are consistent with BMPR2 mutation-driven PAH observed in humans. Given the genetic and physiological similarities of BMPR2(+/-)sheep to humans with heritable PAH, this large animal model will serve as a vital platform for mechanistic molecular studies and will provide a much-needed pre-clinical model for extensive treatment evaluations.
]]></description>
<dc:creator>Datar, S. A.</dc:creator>
<dc:creator>Werry, N.</dc:creator>
<dc:creator>Brown, A. R.</dc:creator>
<dc:creator>Fitzpatrick, D. S.</dc:creator>
<dc:creator>Falade, O.</dc:creator>
<dc:creator>Trott, J. F.</dc:creator>
<dc:creator>Hutchings, R.</dc:creator>
<dc:creator>Amin, E. K.</dc:creator>
<dc:creator>Morgan, J. M.</dc:creator>
<dc:creator>Nawaytou, H.</dc:creator>
<dc:creator>Deutsch, G. H.</dc:creator>
<dc:creator>Johnson, E. G.</dc:creator>
<dc:creator>Gonzales Viera, O. A.</dc:creator>
<dc:creator>Bishop, T. F.</dc:creator>
<dc:creator>Urbino, T.</dc:creator>
<dc:creator>McNabb, B. R.</dc:creator>
<dc:creator>Austin, E. D.</dc:creator>
<dc:creator>Fineman, J. R.</dc:creator>
<dc:creator>Van Eenennaam, A. L.</dc:creator>
<dc:date>2026-02-07</dc:date>
<dc:identifier>doi:10.64898/2026.02.06.704456</dc:identifier>
<dc:title><![CDATA[A Large Animal Model of Heritable Pulmonary Arterial Hypertension UsingGene-edited BMPR2 Sheep]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.05.704029v1?rss=1">
<title>
<![CDATA[
Harnessing Inflammatory Monocytes to Overcome Resistance to Anti-PD-1 Immunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.05.704029v1?rss=1</link>
<description><![CDATA[
BackgroundResistance to immune checkpoint inhibitors represents a major therapeutic challenge, as less than 50% of patients with melanoma achieve long-term response to immune checkpoint inhibitor therapy. One mechanism of acquired resistance involves somatic mutations, such as loss of beta-2 microglobulin (B2m), that enable tumor cells to evade T cell-mediated killing.

MethodsThis study used single-cell RNA-seq, flow cytometry, and ex vivo functional assays to characterize tumor-infiltrating immune cells in antigen presentation-deficient tumors. Tumor-bearing mice were treated with anti-PD-1 or CD40 agonist antibodies and cell depletion or cytokine blocking antibodies to define mechanisms of action. Analysis of published human RNA-seq datasets was performed to dissect the contributions of inflammatory monocytes to patient outcomes.

ResultsWe found an increase in immunosuppressive macrophages in B2m-null tumors. We hypothesized that repolarizing myeloid cells may restore control of tumor growth. Treatment with CD40 agonist antibody, which promotes differentiation of monocytes and macrophages towards a proinflammatory phenotype, reduced tumor growth and improved survival in B2m-null melanoma and colorectal cancer models. Unexpectedly, both CD8+ T cells and NK cells, but not CD4+ T cells, were required for the efficacy of CD40 agonist, even though CD8+ T cells cannot directly recognize antigen presentation-deficient tumor cells. Instead, these lymphocytes control tumor growth via secretion of IFN{gamma}, as depletion of IFN{gamma} inhibited the therapeutic effect of CD40 agonist. IFN{gamma} receptor (Ifngr1) expression was required on host cells, not tumor cells, for CD40 agonist-mediated tumor control. Single-cell analysis identified a distinct population of inflammatory monocytes that were enriched for an IFN{gamma} response signature in CD40 agonist-treated tumors, suggesting that these cells may be important for tumor control. Analysis of human bulk and single-cell RNA-seq datasets demonstrated that an inflammatory monocyte signature derived from our data was associated with improved patient outcomes and response to immune checkpoint inhibitors.

ConclusionsThese data demonstrate that CD8+ T cells contribute to tumor control even in the absence of direct antigen presentation by tumor cells. More broadly, our work suggests that strategies to activate the effector functions of inflammatory monocytes may limit tumor growth and overcome acquired resistance to immune checkpoint inhibitors.
]]></description>
<dc:creator>Zimmerman, M. P.</dc:creator>
<dc:creator>Huang, A. C.</dc:creator>
<dc:creator>Cox, E. K.</dc:creator>
<dc:creator>Al Abosy, R.</dc:creator>
<dc:creator>Chong, W. L.</dc:creator>
<dc:creator>Bastian, A. G.</dc:creator>
<dc:creator>Vietor, K.</dc:creator>
<dc:creator>Choutri, Y.</dc:creator>
<dc:creator>Collier, J.</dc:creator>
<dc:creator>Zhabotynsky, V.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Fung, M.</dc:creator>
<dc:creator>Weiss, S. A.</dc:creator>
<dc:creator>Robitschek, E. J.</dc:creator>
<dc:creator>Lin, J.-R.</dc:creator>
<dc:creator>Vallius, T.</dc:creator>
<dc:creator>Pant, S.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>HUGO, W.</dc:creator>
<dc:creator>Sen, D. R.</dc:creator>
<dc:creator>Haining, W. N.</dc:creator>
<dc:creator>Sharpe, A. H.</dc:creator>
<dc:creator>Miller, B. C.</dc:creator>
<dc:date>2026-02-08</dc:date>
<dc:identifier>doi:10.64898/2026.02.05.704029</dc:identifier>
<dc:title><![CDATA[Harnessing Inflammatory Monocytes to Overcome Resistance to Anti-PD-1 Immunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.05.704044v1?rss=1">
<title>
<![CDATA[
A decline in follicle cell function is a major driver of Drosophila ovarian aging 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.05.704044v1?rss=1</link>
<description><![CDATA[
The ovary is one of the first organs to lose functionality with age. We found that aging of the Drosophila ovary is characterized by an accumulation of phenotypes in the somatic compartment, including failure of the follicle cells to encapsulate germ-cell cysts, an extended S phase, and increased DNA damage. In aged ovaries, follicle encapsulation defects are associated with the lack of a germ-cell cyst checkpoint in early oogenesis. Single-cell RNA sequencing revealed that, across all cell types in the ovary, cells in the follicle lineage have the highest number of differentially expressed genes. Overexpression of Atg8a, a key autophagy machinery gene homologous to mammalian LC3, specifically in follicle cells prevents age-associated decline in the follicle epithelium and loss of reproductive capacity. Collectively, these findings demonstrate that genetic manipulation of a small population of ovarian somatic cells is sufficient to improve both cell-autonomous and non-autonomous features of reproductive aging.
]]></description>
<dc:creator>Wolfgram, E. A.</dc:creator>
<dc:creator>Nystul, T. G.</dc:creator>
<dc:date>2026-02-08</dc:date>
<dc:identifier>doi:10.64898/2026.02.05.704044</dc:identifier>
<dc:title><![CDATA[A decline in follicle cell function is a major driver of Drosophila ovarian aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.08.704658v1?rss=1">
<title>
<![CDATA[
MAMBAxBrain: A Multi-task Neural Framework Linking Brain Functional Dynamics to Individual Fingerprints, Cognitive and Disease States 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.08.704658v1?rss=1</link>
<description><![CDATA[
Functional magnetic resonance imaging (fMRI) contains rich individual, cognitive, and pathological information, yet no universal model exists for multi-task modeling of these dimensions. Here, we introduce MAMBAxBrain, a multi-task neural framework that integrates Mamba architecture with functional connectivity analysis to jointly model the temporal dynamics and spatial coordination of neural activity. MAMBAxBrain achieves high accuracy across four distinct fMRI objectives--brain fingerprinting, cognitive task decoding, reaction time prediction, and schizophrenia classification--consistently outperforming state-of-the-art methods with robust crosssession generalization. Interpretability analyses show that each task engages distinct, biologically plausible circuitry--from higher-order association cortex for identity to subcortical-motor loops for reaction time and disrupted control-sensory connectivity for schizophrenia. These findings inform a longstanding debate: rather than operating through wholly separate or entirely shared systems, the brain preferentially recruits task-specific circuits while retaining common representational structure across functions.
]]></description>
<dc:creator>Xia, Y.</dc:creator>
<dc:creator>Arab, F.</dc:creator>
<dc:creator>Saha, U.</dc:creator>
<dc:creator>Sipes, B.</dc:creator>
<dc:creator>Gooden, G.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Raj, A.</dc:creator>
<dc:date>2026-02-10</dc:date>
<dc:identifier>doi:10.64898/2026.02.08.704658</dc:identifier>
<dc:title><![CDATA[MAMBAxBrain: A Multi-task Neural Framework Linking Brain Functional Dynamics to Individual Fingerprints, Cognitive and Disease States]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.09.704975v1?rss=1">
<title>
<![CDATA[
Systemic hypoxia suppresses solid tumor growth 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.09.704975v1?rss=1</link>
<description><![CDATA[
Local hypoxia is a hallmark of solid tumors and a negative prognostic factor in the progression and treatment of cancer. Here, we showed that systemic hypoxia, in contrast to localized tumor hypoxia, decreases tumor growth in vivo across multiple cancer types and preclinical models. The reduced tumor growth in systemic hypoxia was not explained by hypoglycemia, hypoinsulinemia, or HIF activation. Instead, metabolite profiling in tumors and tumor interstitial fluid revealed extensive perturbations in purine-related metabolites. Stable isotope tracing demonstrated that systemic hypoxia caused tumors to suppress de novo purine synthesis. Furthermore, tumors did not develop resistance to systemic hypoxia therapy, and when used in combination with chemotherapy or immunotherapy, systemic hypoxia dramatically suppressed tumor growth. Finally, we showed that systemic hypoxia can be achieved pharmacologically with the small molecule HypoxyStat. These findings challenge the long-held paradigm of hypoxia as a negative prognostic factor in cancer progression, and they suggest a potential therapeutic role for systemic hypoxia in suppressing solid tumor growth.
]]></description>
<dc:creator>Midha, A. D.</dc:creator>
<dc:creator>Chew, B. T. L.</dc:creator>
<dc:creator>Choi, B. M. H.</dc:creator>
<dc:creator>Suh, J. M.</dc:creator>
<dc:creator>Carpenter, C.</dc:creator>
<dc:creator>Baik, A. H.</dc:creator>
<dc:creator>Joshi, T.</dc:creator>
<dc:creator>Blume, S. Y.</dc:creator>
<dc:creator>Haribowo, A. G.</dc:creator>
<dc:creator>Ruivo, P.</dc:creator>
<dc:creator>Flanigan, W. R.</dc:creator>
<dc:creator>Garg, A.</dc:creator>
<dc:creator>Zhang, D. D.</dc:creator>
<dc:creator>Subramanyam, V.</dc:creator>
<dc:creator>Shuere, R.</dc:creator>
<dc:creator>Seo, Y.</dc:creator>
<dc:creator>VanBrocklin, H.</dc:creator>
<dc:creator>Goodarzi, H.</dc:creator>
<dc:creator>Jain, I. H.</dc:creator>
<dc:date>2026-02-10</dc:date>
<dc:identifier>doi:10.64898/2026.02.09.704975</dc:identifier>
<dc:title><![CDATA[Systemic hypoxia suppresses solid tumor growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.10.704562v1?rss=1">
<title>
<![CDATA[
PRECISE TBI Model Catalog: Increasing Accessibility and Reproducibility in TBI research 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.10.704562v1?rss=1</link>
<description><![CDATA[
Preclinical traumatic brain injury (TBI) research relies on experimental models that vary by mechanism, parameters, surgical procedures, species, strains, and ages, to name a few. While these models are crucial for understanding injury mechanisms and testing therapies, the progress in translating this knowledge to the clinic has been limited. This is in part due to fragmented resources and inconsistent reporting of critical variables. Here, we introduce the PRECISE-TBI model catalog, a centralized, queryable resource that consolidates metadata from published studies. The catalog integrates curated annotations from more than 450 papers, including details such as age, sex, strain, model type, device, and injury parameters. Where available, entries are also linked to protocols and datasets to enhance transparency and reproducibility. The Model Catalog serves as a living resource that enables cross-study comparison, identifies gaps in reporting, and connects the literature to datasets, protocols, device information, and other relevant resources. Analysis of the initial catalog entries revealed gaps in the reporting of device, age, and weight. In contrast, the reporting of sex improved over time, with over 90% of recent studies within the catalog papers reporting sex. Strain was also reported in most studies, with consistent reporting of specificity, especially for the C57 mice substrain. We expect the Model Catalog to serve as a valuable tool to enhance study design and reproducibility in preclinical TBI research while advancing FAIR data principles in the TBI field.
]]></description>
<dc:creator>Surles-Zeigler, M. C.</dc:creator>
<dc:creator>Holmes, L.</dc:creator>
<dc:creator>Sincomb, T.</dc:creator>
<dc:creator>Martone, M. E.</dc:creator>
<dc:creator>Grethe, J.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Dixon, C. E.</dc:creator>
<dc:date>2026-02-12</dc:date>
<dc:identifier>doi:10.64898/2026.02.10.704562</dc:identifier>
<dc:title><![CDATA[PRECISE TBI Model Catalog: Increasing Accessibility and Reproducibility in TBI research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.10.705199v1?rss=1">
<title>
<![CDATA[
Shared somatosensory-motor neural population dynamics track motor recovery after stroke 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.10.705199v1?rss=1</link>
<description><![CDATA[
SO_SCPLOWUMMARYC_SCPLOWAlthough somatosensation is known to be important for precise hand control, how coordinated activity across sensory and motor cortical areas supports hand movements and object interactions is unclear. We simultaneously recorded neural activity from dorsal premotor cortex (PMd) and Area 2 of the primary somatosensory cortex in three rhesus macaques performing a reach-to-grasp task during recovery from a primary motor cortex (M1) lesion. Using dual-area latent factor analysis, we decomposed population activity into cross-area factors (CFs) shared between PMd and Area 2 and within-area factors (WFs) local to each region. Shared CFs tracked hand kinematics closely despite explaining less overall variance. Strikingly, the emergence of long-timescale, stereotyped sensory-motor cross-area dynamics was correlated with recovery of prehension and object grasping, whereas changes in within-area dynamics were not correlated with recovery. These findings suggest that restoration of coordinated sensory-motor population dynamics is important for recovery of skilled hand function after injury.
]]></description>
<dc:creator>Heimbuch, I. S.</dc:creator>
<dc:creator>Khanna, P.</dc:creator>
<dc:creator>Novik, L.</dc:creator>
<dc:creator>Morecraft, R. J.</dc:creator>
<dc:creator>Ganguly, K.</dc:creator>
<dc:date>2026-02-12</dc:date>
<dc:identifier>doi:10.64898/2026.02.10.705199</dc:identifier>
<dc:title><![CDATA[Shared somatosensory-motor neural population dynamics track motor recovery after stroke]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.12.705620v1?rss=1">
<title>
<![CDATA[
Distinct tau filament folds in familial frontotemporal dementia due to the MAPT S305I mutation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.12.705620v1?rss=1</link>
<description><![CDATA[
Frontotemporal lobar degeneration with tau inclusions (FTLD-tau) comprise a class of fatal heterogeneous neurodegenerative diseases. Approximately 10% arise from pathogenic MAPT mutations and often cause severe, early-onset disease with pathology that is distinct yet partially overlapping with sporadic cases. Here, we evaluated post-mortem tissue from a patient with FTLD-tau due to MAPT S305I showing neuropathology most consistent with argyrophilic grain disease (AGD), a prevalent limbic tauopathy of aging. Structures determined by cryo-electron microscopy reveal tau filament folds that differ from those found in sporadic AGD or other tauopathies and feature a 4-layer architecture stabilized by the Ile substitution within its core. Comparative structural analysis reveals conserved motifs are shared among AGD, corticobasal degeneration, and MAPT P301T. A well-defined density stacks along a cationic cleft, indicative of a bound RNA-like polyanion or small-molecule. In vitro analysis shows the S305I mutation promotes fibrilization relative to normal tau. These results demonstrate that MAPT S305I stabilizes a distinct aggregation-prone tau fold that likely contributes to disease pathology and heterogeneity beyond its known splicing defects, and underscore potential limitations of using the most pathologically similar genetic form as a model for sporadic FTLD-tau.
]]></description>
<dc:creator>Pan, H. S.</dc:creator>
<dc:creator>Merz, G. E.</dc:creator>
<dc:creator>Li, A. N.</dc:creator>
<dc:creator>Le, M. Q.</dc:creator>
<dc:creator>Jo, H.</dc:creator>
<dc:creator>Quddus, A.</dc:creator>
<dc:creator>Yung, A.</dc:creator>
<dc:creator>Kormos, R.</dc:creator>
<dc:creator>Melo, A. A.</dc:creator>
<dc:creator>Ramos, E. M.</dc:creator>
<dc:creator>Lago, A. L.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Rosen, H. H.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Gorno-Tempini, M. L.</dc:creator>
<dc:creator>DeGrado, W. F.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Southworth, D. R.</dc:creator>
<dc:date>2026-02-14</dc:date>
<dc:identifier>doi:10.64898/2026.02.12.705620</dc:identifier>
<dc:title><![CDATA[Distinct tau filament folds in familial frontotemporal dementia due to the MAPT S305I mutation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.16.706171v1?rss=1">
<title>
<![CDATA[
Inhibition of the EBF1-ITGB8 Axis in Bone Marrow Niche Ameliorates Hallmarks of Myelofibrosis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.16.706171v1?rss=1</link>
<description><![CDATA[
Fibrotic remodeling of the bone marrow (BM) niche is a characteristic feature of myelofibrosis (MF) that contributes to disease progression. In MF, mesenchymal stromal cells (MSCs) produce excessive amounts of inflammatory cytokines and extracellular matrix, leading to BM fibrosis, impaired blood production, extramedullary hematopoiesis, and progressive BM failure. While the genetic events that initiate MF in hematopoietic cells are well defined, our understanding of the mechanisms responsible for BM fibrosis remains incomplete. Here, we show that transcription factor EBF1 is a key regulator of the fibrotic gene program in mouse and human MSCs. EBF1 is upregulated in pre-fibrotic MSCs, while mice with MSC-specific deletion of Ebf1 exhibit reduced BM fibrosis, decreased expansion of myeloid cells and splenomegaly when transplanted with hematopoietic progenitors harboring the MF driver mutation MPLW515L. Moreover, we identify ITGB8 as an EBF1-regulated gene with therapeutic potential. MF mice treated with ITGB8-neutralizing antibodies or with MSC-specific Itgb8 deletion show reduced disease burden, as indicated by decreased marrow fibrosis, significantly reduced frequencies of MPL mutant cells, and reduced inflammation in the BM. Our data indicate that targeting the EBF1-ITGB8 axis in the MF MSCs may have therapeutic benefits.
]]></description>
<dc:creator>Tsurkan, L.</dc:creator>
<dc:creator>Doute, M.</dc:creator>
<dc:creator>Morchel, N.</dc:creator>
<dc:creator>Konada, L.</dc:creator>
<dc:creator>Mehmood, R.</dc:creator>
<dc:creator>Ling, T.</dc:creator>
<dc:creator>Atakilit, A.</dc:creator>
<dc:creator>Marcellino, B.</dc:creator>
<dc:creator>Hoffman, R.</dc:creator>
<dc:creator>Vogel, P.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:creator>Crispino, J.</dc:creator>
<dc:creator>Derecka, M.</dc:creator>
<dc:date>2026-02-17</dc:date>
<dc:identifier>doi:10.64898/2026.02.16.706171</dc:identifier>
<dc:title><![CDATA[Inhibition of the EBF1-ITGB8 Axis in Bone Marrow Niche Ameliorates Hallmarks of Myelofibrosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.13.705828v1?rss=1">
<title>
<![CDATA[
Age-related Delays in Osteochondral Remodeling of Fracture Healing Illustrated by Mass Spectrometry Imaging 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.13.705828v1?rss=1</link>
<description><![CDATA[
Age-related delays in fracture healing are prevalent and contribute to morbidity and mortality in elderly populations. Clinical and preclinical studies demonstrate that aging is associated with slower and less complete fracture repair characterized by delayed cartilage and bone formation, impaired matrix resorption, and an increased risk of delayed union or non-union. Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI MSI) enables spatially resolved, in situ molecular analysis of proteins directly from murine fracture tissues. We applied collagenase type III (MMP-13) mediated proteolytic digestion to formalin-fixed, paraffin-embedded (FFPE) tibia fracture callus sections harvested 10 days post-tibial fracture from young (3-month-old) and aged (18-month-old) mice to perform spatially resolved proteomic profiling. Spatial MS Imaging revealed pronounced age-dependent differences in extracellular matrix protein composition and remodeling within the fracture callus. We identified upregulation of canonical bone and matrix proteins, including Col1a1 and Col1a2 specifically in the young fracture callus demonstrating advancement into harden callus formation. Conversely, Col2a1 and other soft callus proteins were only seen in the aged callus tissues. Further, protein indicators of tissue state, such as fibronectin (upregulated) and calreticulin (downregulated) were selectively regulated aged tissues, demonstrating a failure for aged tissues to fully progress into harden calluses. Spatial proteomic patterns demonstrated a marked delay in progression from cartilaginous to osseous callus in aged mice, consistent with impaired matrix remodeling during fracture repair. Together, these findings establish spatial MS Imaging based proteomics as a powerful approach to elucidate age-related alterations in fracture healing and to identify molecular regulators of impaired skeletal regeneration.

Lay SummaryUsing spatially-resolved proteomics via mass spectrometry imaging on fracture callus tissues from young and aged mice, we observed delayed healing in aged animals based on the composition of the extracellular matrices. Higher levels of bone specific collagens were detected in young animals, whereas cartilage specific collagens were detected in aged animals at higher levels. Further, detection of novel, non-canonical callus proteins revealed critical transitional steps that are delayed in aged-callus tissues, and these may also contribute to the delayed healing aged animals.
]]></description>
<dc:creator>Schurman, C. A.</dc:creator>
<dc:creator>Chandler, W.</dc:creator>
<dc:creator>Hu, D.</dc:creator>
<dc:creator>Taylor, H.</dc:creator>
<dc:creator>Tao, N.</dc:creator>
<dc:creator>Miclau, T.</dc:creator>
<dc:creator>Angel, P.</dc:creator>
<dc:creator>Marcucio, R.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.13.705828</dc:identifier>
<dc:title><![CDATA[Age-related Delays in Osteochondral Remodeling of Fracture Healing Illustrated by Mass Spectrometry Imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.16.706219v1?rss=1">
<title>
<![CDATA[
Structural modification of oxazolidinone antibiotics alters nascent peptide stalling preference and peptide trajectory through the ribosome 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.16.706219v1?rss=1</link>
<description><![CDATA[
The oxazolidinone antibiotic linezolid binds to the peptidyl transferase center of the ribosome, where it inhibits a subset of peptide bond formation events. This context-specificity of translation inhibition is dictated by the nature of the amino acid at the penultimate position of the nascent peptide. It remains unknown whether this is a general feature of oxazolidinones and whether it can be modulated by their structural alterations. Here, we show that the oxazolidinone tedizolid also inhibits translation in a context-specific manner, but with dramatically altered selectivity, favoring Ile, His, and Gln as the penultimate residues. Delpazolid, which shares the C5 hydroxymethyl moiety with tedizolid, shows a similar preference. Structural analysis of the ribosome with tedizolid and a stalled nascent peptide showed a compacted, helical conformation of the nascent chain induced by the drug. Our findings reveal that stalling preferences of oxazolidinones can be modulated by structural modifications within this antibiotic class.
]]></description>
<dc:creator>Kleinman, J. I.</dc:creator>
<dc:creator>Raskar, T.</dc:creator>
<dc:creator>Klepacki, D.</dc:creator>
<dc:creator>Szal, T.</dc:creator>
<dc:creator>Vazquez-Laslop, N.</dc:creator>
<dc:creator>Mankin, A.</dc:creator>
<dc:creator>Fraser, J. S.</dc:creator>
<dc:creator>Fujimori, D. G.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.16.706219</dc:identifier>
<dc:title><![CDATA[Structural modification of oxazolidinone antibiotics alters nascent peptide stalling preference and peptide trajectory through the ribosome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.17.705849v1?rss=1">
<title>
<![CDATA[
Structural evolution of the MTCH family of mitochondrial insertases 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.17.705849v1?rss=1</link>
<description><![CDATA[
Here we demonstrated that MTCH2 is the defining member of a large family of mitochondrial outer membrane (OM) insertases. The MTCH family is conserved across holozoa and has diverged from the solute carrier 25 transporters. The cryoelectron microscopy structure of the 33 kDa human MTCH2 revealed that evolution of its insertase activity required loss of a transmembrane helix, which created a lipid-accessible hydrophilic groove stabilized by its unique, structured C-terminus. Mutational analyses showed that MTCH insertase activity is attenuated, while experimental structures of hyperactive mutants demonstrated that the hydrophobicity, charge, and size of the residues that line its groove regulated MTCH function. Leveraging the MTCH2 structure, we identified the plant OM insertase, and proposed a universal mechanism for OM insertion across all kingdoms of life.
]]></description>
<dc:creator>Stevens, T. A.</dc:creator>
<dc:creator>Luo, Z.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Hazu, M.</dc:creator>
<dc:creator>Galatis, E. G.</dc:creator>
<dc:creator>Inglis, A. J.</dc:creator>
<dc:creator>Guna, A.</dc:creator>
<dc:creator>Voorhees, R. M.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.17.705849</dc:identifier>
<dc:title><![CDATA[Structural evolution of the MTCH family of mitochondrial insertases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.17.706445v1?rss=1">
<title>
<![CDATA[
In vivo deuteration reveals pronounced variation in myelin lipid turnover rates and reduced myelin renewal with ageing 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.17.706445v1?rss=1</link>
<description><![CDATA[
Myelin turnover is essential for its structural and functional integrity, yet how this particularly lipid-rich membrane is renewed and why it deteriorates with ageing remain unresolved. Combining deuterium oxide administration in mice with high resolution lipidomics, we establish that brain lipid turnover rates are highly heterogeneous, differ by brain region, and depend primarily on lipid class. Half-lives of common glycerophospholipids in purified myelin were under 2 months whereas many sphingolipids exhibited half-lives exceeding 8 months, dependent on acyl chain length and saturation. Myelin sphingolipid and cholesterol replacement rates in the corpus callosum decreased markedly between 3 and 12 months of age, while disrupting lipid trafficking through ApoE ablation preferentially impaired cholesterol turnover and incorporation into myelin. Our results establish that myelin renewal occurs through continual replacement of individual lipid constituents in a manner that depends on lipid class, hydrophobicity, and ApoE-dependent trafficking, and that this process slows significantly with ageing.
]]></description>
<dc:creator>Lee, J. Y.</dc:creator>
<dc:creator>Cai, Y.</dc:creator>
<dc:creator>Westerhausen, M.</dc:creator>
<dc:creator>Michael, J. A.</dc:creator>
<dc:creator>Teo, J. D.</dc:creator>
<dc:creator>Song, H.</dc:creator>
<dc:creator>Watt, G.</dc:creator>
<dc:creator>Ellis, S. R.</dc:creator>
<dc:creator>Don, A. S.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.17.706445</dc:identifier>
<dc:title><![CDATA[In vivo deuteration reveals pronounced variation in myelin lipid turnover rates and reduced myelin renewal with ageing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.17.706468v1?rss=1">
<title>
<![CDATA[
Targeted shedding of extracellular membrane proteins by induced protease recruitment 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.17.706468v1?rss=1</link>
<description><![CDATA[
Extracellular targeted protein degradation has emerged as a promising therapeutic modality to eliminate proteins of interest (POIs) at the cell surface, by using bifunctional molecules to recruit natural recycling receptors or membrane-bound E3 ligases that redirect POIs to the lysosome. Another natural mechanism involves extracellular proteases that cleave and shed extracellular domains. Here, we exploit this endogenous mechanism by engineering bispecific antibody Shedders, that recruit a classic sheddase ADAM10 to POIs, inducing selective ectodomain shedding. We first targeted the immune checkpoint receptor LAG-3 and observed robust depletion of surface LAG-3 accompanied by accumulation of soluble LAG-3 fragments in both engineered cell lines and primary human T cells. Using biochemical and imaging assays, we confirmed that this antibody-induced shedding is restricted to extracellular protease activity and occurs independently of lysosomal trafficking. Notably, induced shedding of LAG-3 on activated primary T cells partially alleviated inhibitory signaling and reinvigorated IFN{gamma} secretion. We extended the scope of induced shedding by developing Shedders that recognize synthetic epitope-tags that enabling rapid assessment of substrate compatibility across diverse targets. Using this platform, we identified multiple immune modulatory cell-surface receptors, including IL6R, CD62L and MIC-A that can be targeted for shedding. In summary, this work establishes a new paradigm for targeted extracellular proteolysis and expands the toolkit for studying extracellular proteolysis with potential therapeutic benefit.
]]></description>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Zhao, F.</dc:creator>
<dc:creator>Miao, K.</dc:creator>
<dc:creator>Peters-Clarke, T. M.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Ganjave, S. D.</dc:creator>
<dc:creator>Vazquez-Maldonado, A. L.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Kumru, K.</dc:creator>
<dc:creator>Jumaa, H.</dc:creator>
<dc:creator>Leung, K. K.</dc:creator>
<dc:creator>Wells, J. A.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.17.706468</dc:identifier>
<dc:title><![CDATA[Targeted shedding of extracellular membrane proteins by induced protease recruitment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.17.706477v1?rss=1">
<title>
<![CDATA[
Reconstitution of Ras-PI3Kγ membrane communication and feedback using light-induced signaling inputs 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.17.706477v1?rss=1</link>
<description><![CDATA[
Biochemical reactions involving phosphatidylinositol phosphate (PIP) lipids and small GTPases serve essential roles in signal transduction immediately downstream of cell surface receptor activation. Interconnected positive and negative feedback loops link these distinct classes of reactions and allow for the rapid establishment of the steep and precisely oriented intracellular activity gradients that are hallmarks of cell polarity. Although genetic approaches have identified numerous molecules that potentially regulate feedback between GTPases and PIP lipids in cells, it is unclear how different types of feedback modulate the spatiotemporal dynamics of membrane signaling reactions. Here, we reconstitute communication and feedback between Ras GTPase and phosphatidylinositol 3-kinase gamma (PI3K{gamma})-mediated generation of PIP3 on supported membranes. We employ light-induced membrane recruitment of a guanine nucleotide exchange factor (GEF) to rapidly and reversibly shift steady-state conditions and monitor corresponding Ras activation and PIP3 production in time and space. Alone, the Ras-PI3K{gamma} module exhibits weak and transient activation due to global inhibitors. However, the introduction of GEF-mediated positive feedback drives local amplification of Ras activation and PIP3 generation, resulting in a wave of activity that propagates across the membrane surface. Spatial coupling between activated Ras and PIP3 molecules depends on their respective diffusion coefficients and the species involved in the feedback circuit. This work identifies activation thresholds, membrane diffusion, and feedback architecture as key factors that determine how small GTPases and PIP lipid modifying enzymes amplify membrane signaling reactions in the presence of global inhibitors.
]]></description>
<dc:creator>Doerr, S.</dc:creator>
<dc:creator>Olavarrieta-Colasurdo, A.</dc:creator>
<dc:creator>Hansen, S. D.</dc:creator>
<dc:date>2026-02-18</dc:date>
<dc:identifier>doi:10.64898/2026.02.17.706477</dc:identifier>
<dc:title><![CDATA[Reconstitution of Ras-PI3Kγ membrane communication and feedback using light-induced signaling inputs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.18.706703v1?rss=1">
<title>
<![CDATA[
Unique mineralization pattern revealed in TBCK syndrome mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.18.706703v1?rss=1</link>
<description><![CDATA[
TBCK syndrome is a severe degenerative leukoencephalopathy with multisystem involvement. Neurodevelopmental, craniofacial, and pulmonary challenges are among the topmost effects on these children. TBCK has been implicated in endo-lysosomal regulation, RNA transport, and mTOR-associated pathways, all of which are critical for the development of mineralized tissue. Although craniofacial abnormalities can be clinically apparent, conventional imaging approaches may overlook subtle defects in mineral quality.

Here, we apply our multimodal framework to investigate the mineralization of enamel, dentin, and alveolar bone in a Tbck knockout mouse model. This is the first time our multimodal framework will be applied to a genetic condition. Using micro-computed tomography (microCT), histology, nanoindentation, energy-dispersive spectroscopy, and Raman spectroscopy, we identify tissue- and stage-dependent mineral effects undetected by microCT alone. Tbck loss resulted in differences in enamel and dentin element compositions as early as secretory and transition stages, while mechanical properties remained undetected until maturation stage.

Notably, Tbck knockout enamel exhibited reduced calcium and phosphorus content, along with increased carbon content during early mineralization, consistent with the retained organic matrix. Additionally, marked and opposing alterations in magnesium and iron levels began at the secretory stage. Together, these findings define a previously unrecognized mineralization signature associated with TBCK deficiency and establish multimodal hard-tissue analysis as a sensitive approach for detecting early craniofacial phenotypes in rare genetic disorders.
]]></description>
<dc:creator>Katsura, K. A.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Didziokas, M.</dc:creator>
<dc:creator>Badt, N. Z.</dc:creator>
<dc:creator>Dougherty, S.</dc:creator>
<dc:creator>Vining, K. H.</dc:creator>
<dc:creator>Bhoj, E. J.</dc:creator>
<dc:date>2026-02-20</dc:date>
<dc:identifier>doi:10.64898/2026.02.18.706703</dc:identifier>
<dc:title><![CDATA[Unique mineralization pattern revealed in TBCK syndrome mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.19.706904v1?rss=1">
<title>
<![CDATA[
Autoantibody Repertoires in Healthy Individuals Vary According to HLA Class II Genotype 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.19.706904v1?rss=1</link>
<description><![CDATA[
The link between HLA genotype and the presence of pathogenic autoantibodies has previously been established across different autoimmune disorders. However, the functional process linking specific antibodies to specific HLA remains unclear. Additionally, autoantibodies - usually associated with autoimmune disease - are also present in healthy individuals. To this end, we sought to determine the spectrum of self-antigen antibody specificity (the "autoreactome") in healthy individuals, stratified by HLA genotypes. We utilized Phage ImmunoPrecipitation Sequencing (PhIP-Seq), a programmable phage display for interrogation of antibody specificity, which encompasses over 700,000 peptides tiled across the entire human proteome. Serum from 741 donors without diagnosed autoimmune disease were grouped by their five homozygous HLA-DRB1 genotypes and analyzed for differences in autoreactivity profiles. We applied a custom filter to the PhIP-Seq normalized data and obtained a set of enriched peptides for each HLA-DRB1 genotype. We found that the autoreactome in healthy individuals with different HLA-DRB1 genotypes are generally distinct. Binary logistic regression successfully identified whether a sample belongs to a specific HLA-DRB1 genotype (HLA-DRB1*01, HLA-DRB1*03, HLA-DRB1*04, HLA-DRB1*07, and HLA-DRB1*15) with the following accuracies: 96%, 92%, 90%, 94%, and 90%, and multinomial models predicted HLA-DRB1 genotype with up to 90% accuracy. Finally, gene-level analysis suggests that individuals with specific HLA autoimmune risk alleles may harbor potentially pathogenic autoantibodies in the absence of, or prior to the establishment of, overt disease. Our analysis demonstrates that autoreactivity profiles in healthy people vary according to HLA class II genotype, and may provide insight into the pathological processes associated with development of autoimmune and other immune-mediated diseases.
]]></description>
<dc:creator>Suseno, R.</dc:creator>
<dc:creator>Boquett, J. A.</dc:creator>
<dc:creator>Dandekar, R.</dc:creator>
<dc:creator>Ituarte, T.</dc:creator>
<dc:creator>Alvarenga, B. D.</dc:creator>
<dc:creator>Vierra-Green, C.</dc:creator>
<dc:creator>Spellman, S.</dc:creator>
<dc:creator>Maiers, M.</dc:creator>
<dc:creator>Wilson, M.</dc:creator>
<dc:creator>Hollenbach, J. A.</dc:creator>
<dc:date>2026-02-20</dc:date>
<dc:identifier>doi:10.64898/2026.02.19.706904</dc:identifier>
<dc:title><![CDATA[Autoantibody Repertoires in Healthy Individuals Vary According to HLA Class II Genotype]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.19.706910v1?rss=1">
<title>
<![CDATA[
Induction and regulation of a reversible form of suspended animation in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.19.706910v1?rss=1</link>
<description><![CDATA[
Suspended animation, a state of extreme quiescence with microscopically invisible movement and development, is a remarkable yet poorly understood stress resilience strategy in animals. Here, we describe a newly discovered form of suspended animation inducible by high-population density in isosmotic liquids in C. elegans throughout larval development and adulthood. Transcriptomic, metabolomic and live-cell activity reporter imaging analyses reveal striking molecular and cellular landscape changes by such liquid-induced suspended animation (LISA), including remodeling of gene expression programs, energy metabolites, lysosomal and mitochondrial morphology. Genetic screens identify mutants with altered stress responses and survival against LISA. While key endo-lysosomal regulators promote survival during LISA, organelle remodeling and a neuronal axis via downstream neuropeptide and cAMP/PKA signaling orchestrate behavioral awakening from LISA. Our findings define a facile paradigm for reversible SA, providing a powerful model system to uncover key molecular and cellular mechanisms governing an extreme case of reversible life arrest and dormancy.
]]></description>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Catrow, J. L.</dc:creator>
<dc:creator>Pearce, Q.</dc:creator>
<dc:creator>Ji, Z.</dc:creator>
<dc:creator>Yang, S. W.</dc:creator>
<dc:creator>Balakrishnan, A.</dc:creator>
<dc:creator>Cox, J.</dc:creator>
<dc:creator>Ma, D. K.</dc:creator>
<dc:date>2026-02-20</dc:date>
<dc:identifier>doi:10.64898/2026.02.19.706910</dc:identifier>
<dc:title><![CDATA[Induction and regulation of a reversible form of suspended animation in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.19.706929v1?rss=1">
<title>
<![CDATA[
A universal platform for simultaneous TCRα/β removal enables safer and more potent TCR therapies and autoimmune modeling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.19.706929v1?rss=1</link>
<description><![CDATA[
Adoptive T-cell therapies using tumour-specific T-cell receptors (TCRs) are limited by competition with endogenous receptors, which impairs efficacy and poses risks of off-target autoreactivity. Here we present a CRISPR-based platform that completely and selectively eliminates both endogenous TCR- and -{beta} chains without affecting introduced transgenic TCRs, irrespective of codon optimization. This approach achieves >90% deletion efficiency in Jurkat and primary human T cells, markedly enhancing the expression, pairing fidelity, and functional potency of transgenic receptors. Using a clinically relevant HLA-A*02:01-restricted DMF5 TCR, we show that dual TCR ablation boosts antigen-specific activation and cytotoxicity in vitro and significantly enhances tumor clearance in vivo in human immune system (HIS) mice, while preventing graft-versus-host disease (GVHD). Targeted locus amplification revealed that CRISPR-induced double-strand breaks did not alter lentiviral integration profiles, confirming genomic safety. Extending this approach to four insulin-reactive TCRs demonstrated that removal of endogenous receptors increased transduction efficiency and functional activity, with one (1E6) showing selective activation and infiltration of stem cell-derived islet grafts (SC-islets) in vivo. This study establishes a universal, safe, and scalable genome-editing platform for generating functionally precise human T cells. By integrating cancer immunotherapy and autoimmune disease modelling within a single framework, it provides a strong preclinical rationale for dual endogenous TCR removal as a route to improved specificity, safety, and therapeutic efficacy in TCR-based cell therapies.
]]></description>
<dc:creator>Zanetti, G.</dc:creator>
<dc:creator>Legut, M.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>Fathi, F.</dc:creator>
<dc:creator>Suek, N.</dc:creator>
<dc:creator>Teteloshvili, N.</dc:creator>
<dc:creator>Li, H. W.</dc:creator>
<dc:creator>Ding, X.</dc:creator>
<dc:creator>Traum, D.</dc:creator>
<dc:creator>Kaestner, K.</dc:creator>
<dc:creator>Hoang, R. E.</dc:creator>
<dc:creator>Bremer, E.</dc:creator>
<dc:creator>Sewell, A. K.</dc:creator>
<dc:creator>Parent, A. V.</dc:creator>
<dc:creator>Creusot, R. J.</dc:creator>
<dc:creator>Sykes, M.</dc:creator>
<dc:creator>Khosravi Maharlooei, M.</dc:creator>
<dc:date>2026-02-20</dc:date>
<dc:identifier>doi:10.64898/2026.02.19.706929</dc:identifier>
<dc:title><![CDATA[A universal platform for simultaneous TCRα/β removal enables safer and more potent TCR therapies and autoimmune modeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.20.706984v1?rss=1">
<title>
<![CDATA[
Acid sphingomyelinase inhibition restores RPE homeostasis and photoreceptor function in preclinical Stargardt macular degeneration models 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.20.706984v1?rss=1</link>
<description><![CDATA[
Stargardt disease, which destroys central high-resolution vision in over 2 million people globally, lacks effective therapies. The primary site of damage in Stargardt disease is the retinal pigment epithelium (RPE), which safeguards photoreceptor health and function. Progressive loss of RPE integrity precedes visual deficits, yet insight into mechanisms driving RPE dysfunction and how this influences disease pathogenesis remains elusive. Here, we addressed this in cell-based and pigmented Abca4-/- Stargardt mice models using super-resolution imaging, bioinformatics, and biochemical approaches. We show that ceramide accumulation induced by bisretinoid-mediated overactivation of acid sphingomyelinase (ASM) in Abca4-/- RPE selectively disrupts Rab GTPases and ESCRT machinery involved in apical membrane trafficking and small extracellular vesicle (EV) biogenesis. Consequently, connexin 43 (Cx43) is misrouted from cell-cell junctions into EVs that are released apically by the RPE. This compromises RPE integrity and promotes subretinal immune cell recruitment, leading to photoreceptor dysfunction. Pharmacological ASM inhibition normalizes EV biogenesis and restores Cx43 localization. Decreasing RPE ceramide safeguards RPE structural integrity, limits subretinal microglia, and improves visual function in Abca4-/- mice. This study underscores the importance of the RPE as a communication hub in the retina and identifies ASM as a potential therapeutic target to prevent progressive vision loss.
]]></description>
<dc:creator>Germer, C. J.</dc:creator>
<dc:creator>Williams, S.</dc:creator>
<dc:creator>Fernandes, V.</dc:creator>
<dc:creator>Espinosa Lima, R.</dc:creator>
<dc:creator>La Cunza, N. R.</dc:creator>
<dc:creator>Tan, L. X.</dc:creator>
<dc:creator>Panda, S. R.</dc:creator>
<dc:creator>Iorio, E.</dc:creator>
<dc:creator>Elahi, F.</dc:creator>
<dc:creator>Lakkaraju, A.</dc:creator>
<dc:date>2026-02-22</dc:date>
<dc:identifier>doi:10.64898/2026.02.20.706984</dc:identifier>
<dc:title><![CDATA[Acid sphingomyelinase inhibition restores RPE homeostasis and photoreceptor function in preclinical Stargardt macular degeneration models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.21.706873v1?rss=1">
<title>
<![CDATA[
The Untangle Challenge for accurate ensemble models 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.21.706873v1?rss=1</link>
<description><![CDATA[
We report the discovery of a new class of local minima that has severely limited the accuracy of macromolecular models. Termed density misfit barrier traps, these minima explain much of the poor fit between macromolecular models and experimental data relative to that of smaller molecules: not just high R factors, but distorted chemical geometry. We postulated that proteins exist as an ensemble of conformations that each have good geometry, but refinement algorithms have been unable to converge to them due to a tangling phenomenon arising from these traps. To demonstrate, a synthetic ground truth data set was generated, consisting of a 2-member ensemble with excellent geometry. A series of starting models, each trapped in increasingly difficult local minima, were prepared, a unified validation score defined, and an open Challenge issued. This Challenge inspired algorithms for escaping such traps, and new programs have been released that are expected to substantially improve the accuracy of macromolecular ensemble models.

SynopsisA synthetic 2-member conformational ensemble of a small protein and corresponding electron density data was generated to demonstrate how topological local minima hinder simultaneous agreement with density data and chemical geometry restraints in conventional structure refinement.
]]></description>
<dc:creator>Hopkins, M. S.</dc:creator>
<dc:creator>Terwilliger, T. C.</dc:creator>
<dc:creator>Afonine, P.</dc:creator>
<dc:creator>Ginn, H. M.</dc:creator>
<dc:creator>HOLTON, J. M.</dc:creator>
<dc:date>2026-02-22</dc:date>
<dc:identifier>doi:10.64898/2026.02.21.706873</dc:identifier>
<dc:title><![CDATA[The Untangle Challenge for accurate ensemble models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.23.707602v1?rss=1">
<title>
<![CDATA[
Title: Optogenetic WNT signaling drives germ layer self-organization in a human gastruloid model 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.23.707602v1?rss=1</link>
<description><![CDATA[
In vitro stem cell models of human gastrulation have been an advance for developmental biology, though elucidating mechanisms of germ layer formation remains challenging. While investigating whether spatially-patterned signaling is required for germ layer formation, we tested a "salt-and-pepper" signaling strategy in which WNT was optogenetically activated in a subset of human pluripotent stem cells (hPSC) uniformly mixed into an aggregate. Following mesendodermal specification, WNT-activated cells spatially segregated into a hemisphere, then underwent further differentiation and organization into mesoderm and endoderm. RNAseq-based lineage analysis revealed that WNT activation non-autonomously induced TGF{beta}/BMP signaling, leading to robust emergence of an anterior visceral endoderm-like population that patterned adjacent neural and mesendodermal fates. Transcriptional profiles and trajectories closely mirrored those observed during human gastrulation. Moreover, TGF{beta} or cadherin perturbation disrupted germ layer formation or spatial organization, respectively. This simple model thus enables mechanistic dissection of complex human lineage specifications and organization during gastrulation.
]]></description>
<dc:creator>Schaffer, D. V.</dc:creator>
<dc:creator>Johnson, H. J.</dc:creator>
<dc:creator>McMullin, D. M.</dc:creator>
<dc:creator>Zimmermann, J.</dc:creator>
<dc:creator>Kim, C.</dc:creator>
<dc:creator>Repina, N.</dc:creator>
<dc:creator>Bhalerao, R.</dc:creator>
<dc:creator>Nowakowski, T.</dc:creator>
<dc:date>2026-02-25</dc:date>
<dc:identifier>doi:10.64898/2026.02.23.707602</dc:identifier>
<dc:title><![CDATA[Title: Optogenetic WNT signaling drives germ layer self-organization in a human gastruloid model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.24.707563v1?rss=1">
<title>
<![CDATA[
Benzoxaboroles are structurally unique binders of eukaryotic translation initiation factor 4E 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.24.707563v1?rss=1</link>
<description><![CDATA[
Benzoxaboroles offer unusual reactivity and protein recognition for the development of small molecule drugs. Despite this potential, they are uncommon in drug discovery or in large fragment screening libraries. We synthesized a small series of structurally related benzoxaboroles containing a diazirine/alkyne tag to enable in-cell photoaffinity labeling (PAL) experiments. A subset of this library was found to have high selectivity for eukaryotic translation initiation factor 4E (eIF4E). The benzoxaborole-eIF4E interaction was found to be stereoselective in nature and competitive with the 7-methylguanosine cap of mRNA. Site of labeling experiments revealed that the benzoxaborole fragment interacts with the cap binding pocket of eIF4E. In silico modeling of the modified protein suggests that H-bonding interactions between the main chain of Trp102 and the side chain of Asn155 to the amide carbonyl and anionic boronate of the benzoxaborole, respectively, drive affinity for this challenging to drug pocket.
]]></description>
<dc:creator>Combs, J. B.</dc:creator>
<dc:creator>Peacock, D. M.</dc:creator>
<dc:creator>Craven, G. B.</dc:creator>
<dc:creator>Jung, S.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Le, S. M.</dc:creator>
<dc:creator>Taunton, J.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:date>2026-02-25</dc:date>
<dc:identifier>doi:10.64898/2026.02.24.707563</dc:identifier>
<dc:title><![CDATA[Benzoxaboroles are structurally unique binders of eukaryotic translation initiation factor 4E]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.24.707611v1?rss=1">
<title>
<![CDATA[
CBASS limits bacteriophage production while maintaining cell viability in Pseudomonas aeruginosa 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.24.707611v1?rss=1</link>
<description><![CDATA[
CBASS is an immune pathway that recognizes phage infection and generates cyclic nucleotide signals, which directly activate effectors that stop phage replication. Membrane-acting effectors are proposed to induce cell death to prevent phage replication; however, this mechanism has not been assessed with endogenous expression levels of the effector. We therefore sought to determine the cell viability outcomes of the CBASS phospholipase effector (CapV) upon activation with 3,3-cGAMP signals in Pseudomonas aeruginosa. Here, we surprisingly observe that constitutive 3,3-cGAMP signaling from the synthase (CdnA) enables robust cell growth and viability while effectively abolishing phage production in a CapV-dependent manner. Exogenous 3,3-cGAMP also enhances CBASS anti-phage activity and cell growth. Moreover, constitutive activation of the CapV effector induces no cell fitness cost, and blocks replication of many, but not all, phages. This demonstrates that a cyclic nucleotide-activated CBASS effector possess a degree of phage specificity that has been previously overlooked. When CBASS is active, phage transcription and initiation of DNA replication proceed normally, but phages do not reach maximum DNA levels and fewer mature virions are produced. Based on these findings, we propose that CapV interferes with the early stages of phage capsid assembly at the cell membrane and resultantly disrupts DNA packaging. Collectively, we demonstrate that a successful CBASS response antagonizes a late-stage of the phage replication cycle while maintaining cell viability.
]]></description>
<dc:creator>Huiting, E.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Shmidov, E.</dc:creator>
<dc:creator>Musunuri, S.</dc:creator>
<dc:creator>Gitai, Z.</dc:creator>
<dc:creator>Bondy-Denomy, J.</dc:creator>
<dc:date>2026-02-25</dc:date>
<dc:identifier>doi:10.64898/2026.02.24.707611</dc:identifier>
<dc:title><![CDATA[CBASS limits bacteriophage production while maintaining cell viability in Pseudomonas aeruginosa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.23.704410v1?rss=1">
<title>
<![CDATA[
Bradykinin Contributes to Vasogenic Edema in Murine Experimental Cerebral Malaria 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.23.704410v1?rss=1</link>
<description><![CDATA[
Cerebral malaria (CM) due to Plasmodium falciparum (Pf) infection is a major cause of death in African children. Bradykinin (BK) is a mediator of vasogenic edema. It could contribute to the pathogenesis of central nervous system malaria in Kenyan children and P. berghei ANKA (PbA) infected C57BL/6J mice with experimental cerebral malaria. Cleaved plasma high molecular weight kininogen (cHK) is a marker for prior BK release. 40% of children with central nervous system malaria had plasma cHK versus 18% of children with uncomplicated malaria. Wild-type PbA-infected mice had circulating plasma cHK, elevated BK levels, and reduced HK and prekallikrein levels. HK null (Kng1-/-), combined BK B1 and B2 receptor null (Bdkrb1-/- / Bdkrb2-/-), BK B2 (Bdkrb2-/-) or BK B1 (Bdkrb1-/-) receptor null mice were protected from neurologic deterioration and brain edema compared to wild-type mice. F12-/-mice were not protected from neurological deterioration.

Prekallikrein null (Klkb1-/-), prolylcarboxypeptidase hypomorphs (Prcpgt/gt), and brain endothelial cell conditional knockout of PRCP (Prcpfl/fl Cre) mice had reduced neurologic deterioration and brain edema. Adjuvant plasma kallikrein inhibition combined with artesunate treatment of PbA-infected mice reversed neurologic deterioration and brain edema and prolonged survival relative to artesunate alone. BK-induced vasogenic edema contributes to human and murine CM.
]]></description>
<dc:creator>Pinheiro, A. d. S.</dc:creator>
<dc:creator>Teixeira, D. E.</dc:creator>
<dc:creator>Silva-Aguiar, R. P.</dc:creator>
<dc:creator>Shim, Y. J.</dc:creator>
<dc:creator>Merkulova, A.</dc:creator>
<dc:creator>Silbak, S.</dc:creator>
<dc:creator>Skomorovska-Prokvolit, Y.</dc:creator>
<dc:creator>Midem, D.</dc:creator>
<dc:creator>Ogolla, S.</dc:creator>
<dc:creator>Burckhardt, B. B.</dc:creator>
<dc:creator>Gangnus, T.</dc:creator>
<dc:creator>Scharfstein, J.</dc:creator>
<dc:creator>Caruso-Neves, C.</dc:creator>
<dc:creator>McCarty, O. J.</dc:creator>
<dc:creator>Gailani, D.</dc:creator>
<dc:creator>Bader, M.</dc:creator>
<dc:creator>Rosenthal, P.</dc:creator>
<dc:creator>Dent, A. E.</dc:creator>
<dc:creator>Janse, C. J.</dc:creator>
<dc:creator>McCrae, K.</dc:creator>
<dc:creator>Pinheiro, A. A. d. S.</dc:creator>
<dc:creator>Kazura, J. W.</dc:creator>
<dc:creator>Schmaier, A. H.</dc:creator>
<dc:date>2026-02-26</dc:date>
<dc:identifier>doi:10.64898/2026.02.23.704410</dc:identifier>
<dc:title><![CDATA[Bradykinin Contributes to Vasogenic Edema in Murine Experimental Cerebral Malaria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.25.707725v1?rss=1">
<title>
<![CDATA[
Unified function of FACT in mammalian chromatin replication and transcription, dissolving and restoring nucleosomes to counteract genome aggregation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.25.707725v1?rss=1</link>
<description><![CDATA[
Nucleosomes with their associated modifications underlie genome organisation and regulation. Replication and transcription require nucleosome disassembly to access the DNA template. How this is orchestrated without jeopardizing chromatin function remains unknown. Here, we reveal a general, global requirement of the histone chaperone FACT in mammalian replication, transcription and chromatin maintenance. Upon acute FACT depletion, replisome and RNA polymerase progression is halted genome-wide and chromatin structure in their wake collapses with reduced nucleosome occupancy, irregular spacing and intermediate assemblies. Chromatin states deteriorate as modified histones are lost due to a lack of histone recycling. Chromatin fiber disorder further manifests in the 3D genome, triggering active genes to coalesce in aberrant microcompartments. This establishes a unifying mechanistic basis for mammalian chromatin replication and transcription, with FACT mediating nucleosome disruption and re-assembly and thereby guarding against spurious chromatin aggregation. Nucleosome organization can therefore dynamically regulate genome architecture.
]]></description>
<dc:creator>Frater, R. C. M.</dc:creator>
<dc:creator>Jensen, R. R.</dc:creator>
<dc:creator>Olle Lopez, J.</dc:creator>
<dc:creator>Flury, V.</dc:creator>
<dc:creator>van den Berg, J.</dc:creator>
<dc:creator>Lunow Bournonville, A.</dc:creator>
<dc:creator>Alcaraz, N.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Richter, H.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Du, Q.</dc:creator>
<dc:creator>van Oudenaarden, A.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Krietenstein, N.</dc:creator>
<dc:creator>Groth, A.</dc:creator>
<dc:date>2026-02-26</dc:date>
<dc:identifier>doi:10.64898/2026.02.25.707725</dc:identifier>
<dc:title><![CDATA[Unified function of FACT in mammalian chromatin replication and transcription, dissolving and restoring nucleosomes to counteract genome aggregation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.26.705527v1?rss=1">
<title>
<![CDATA[
Protein-guided RNA barcoding links transcriptomes to synaptic architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.26.705527v1?rss=1</link>
<description><![CDATA[
Mammalian brain function relies on the precise synaptic architecture of diverse cell types, yet scalable methods for linking a neurons transcriptomic profile to its neuroanatomy remain limited. We present Synapse-seq, an in vivo strategy in which cell-identifying barcoded mRNAs are routed to subcellular compartments via targeting proteins and detected by single-cell and spatial genomics. Using AAV delivery for minimal perturbation of gene expression, we directed barcodes to presynaptic terminals (via synaptophysin) in four distinct circuits, or to postsynaptic sites (via nanobodies to endogenous PSD95) of hippocampal excitatory neurons. In the mouse primary visual cortex, presynaptic Synapse-seq recovered known long-range projections and discovered cortical layer subtypes with distinct thalamic innervation. In the anterior cortex, we elucidated simple topographic rules of corticostriatal innervation: intratelencephalic neurons followed a continuous depth-to-target gradient, while extratelencephalic neurons exhibited striatal collaterals that spatially correlated with medullary innervation. Finally, postsynaptic barcoding of excitatory neurons revealed cell type-specific variation in dendritic architectures across and within hippocampal subfields. These data establish Synapse-seq as a versatile, genomics-based approach for the integrated definition of molecular identity and synaptic organization across mammalian brains.
]]></description>
<dc:creator>Urke, A.</dc:creator>
<dc:creator>Dolan, M.-J.</dc:creator>
<dc:creator>Silverman, J.</dc:creator>
<dc:creator>Kim, M. T.</dc:creator>
<dc:creator>Pineda, J.</dc:creator>
<dc:creator>Garcia, S.</dc:creator>
<dc:creator>Luu, J.</dc:creator>
<dc:creator>Buckley, A.</dc:creator>
<dc:creator>Kumar, V.</dc:creator>
<dc:creator>Zhao, B.</dc:creator>
<dc:creator>Chan, K.</dc:creator>
<dc:creator>Nadaf, N.</dc:creator>
<dc:creator>Balderrama, K. S.</dc:creator>
<dc:creator>Arnold, D. B.</dc:creator>
<dc:creator>Stevens, B.</dc:creator>
<dc:creator>Deverman, B. E.</dc:creator>
<dc:creator>Macosko, E. Z.</dc:creator>
<dc:date>2026-02-27</dc:date>
<dc:identifier>doi:10.64898/2026.02.26.705527</dc:identifier>
<dc:title><![CDATA[Protein-guided RNA barcoding links transcriptomes to synaptic architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.26.708346v1?rss=1">
<title>
<![CDATA[
Power is a major confounder in the analysis of cross-ancestry 'portability' in human eQTLs 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.26.708346v1?rss=1</link>
<description><![CDATA[
The phenotypic effects of germline variants are often mediated through gene regulation. Expression quantitative trait loci (eQTLs) are genetic variants associated with changes in gene expression. Understanding how eQTLs vary across populations is essential for characterising the genetic and regulatory drivers of trait diversity. Meta-analysing eQTL studies from multiple populations enables more robust detection of eQTLs and can reveal regulatory mechanisms shaped by population-specific environmental or ancestry-related factors. However, across the multi-ancestry eQTL literature, a wide range of methods have been used to quantify eQTL portability across ancestry groups. Because different studies employ different portability metrics, it is challenging to form a coherent view of the regulatory landscape across populations. In this work, we analyse eQTL summary statistics from ten datasets matched on tissue type and sequencing technology. We compare portability metrics used previously and show that they can yield markedly different patterns of apparent regulatory conservation or divergence. We then examine the statistical determinants of portability across metrics and demonstrate that sample size, minor allele frequency, and linkage disequilibrium are major drivers of the observed differences in eQTL portability across studies. These findings highlight that differences in statistical power stemming from factors such as population size and allele frequency must be accounted for when evaluating eQTL portability. To address this issue, we introduce a new approach designed to correct for these factors when calling eQTL portability. Finally, we show that empirical Bayes multivariate adaptive shrinkage provides a powerful framework for meta-analysing multiple eQTL studies, with the ability to pool signals across populations to produce more robust effect-size estimates within each population.
]]></description>
<dc:creator>Gibbs, P. M.</dc:creator>
<dc:creator>Beasley, I. J.</dc:creator>
<dc:creator>Del Azodi, C. B.</dc:creator>
<dc:creator>McCarthy, D. J.</dc:creator>
<dc:creator>Gallego Romero, I.</dc:creator>
<dc:date>2026-02-27</dc:date>
<dc:identifier>doi:10.64898/2026.02.26.708346</dc:identifier>
<dc:title><![CDATA[Power is a major confounder in the analysis of cross-ancestry 'portability' in human eQTLs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.27.708594v1?rss=1">
<title>
<![CDATA[
Differential Methylation by Early Life Adversity in the Future of Families Child Wellbeing Study 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.27.708594v1?rss=1</link>
<description><![CDATA[
Early life adversity (ELA) has a well-established link to mental health disorders later in life, yet the molecular mechanisms behind this relationship are incompletely understood. The Future of Families Child Wellbeing Study (FFCWS) provides an opportunity to examine experience encoding in the genome through functional changes in DNA methylation (DNAm) in a cohort enriched in subjects exposed to ELA. We investigated epigenome-wide differences in DNAm across thirteen early-life exposures in salivary samples from FFCWS participants. We calculated differential methylation associations with disease-linked genetic variants, evaluated tissue-specific gene expression, and assessed the persistence of DNAm changes from ages 9 to 15 years. Using data from the mSTARR-seq assay, we characterized methylation-dependent regulatory activity. Differential methylation in the FFCWS validated prior results and identified new genomic regions associated with child adversity. Differential methylation occurs in genomic regions likely to impact gene expression, and affected genes are expressed in disease-relevant tissues. We also identified association of genetic variants associated with downstream disorders near differential methylation, including depression, alcohol and substance use, and anxiety disorders. Overall, cumulative ELA is associated with specific DNAm changes, functional regulation, and persistence over time. Our findings indicate that ELA-associated differential methylation in the FFCWS does not simply occur at random, but in genomic regions that are functional. Our results support the conclusion that altered DNAm represents a biological link between early life experience and later health outcomes.
]]></description>
<dc:creator>Dumas Ang, S.</dc:creator>
<dc:creator>Chin, S.</dc:creator>
<dc:creator>Schneper, L. M.</dc:creator>
<dc:creator>Johnston, R. A.</dc:creator>
<dc:creator>Koss, K. J.</dc:creator>
<dc:creator>Mitchell, C.</dc:creator>
<dc:creator>Notterman, D. A.</dc:creator>
<dc:creator>Engelhardt, B. E.</dc:creator>
<dc:creator>Pena, C. J.</dc:creator>
<dc:date>2026-02-28</dc:date>
<dc:identifier>doi:10.64898/2026.02.27.708594</dc:identifier>
<dc:title><![CDATA[Differential Methylation by Early Life Adversity in the Future of Families Child Wellbeing Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.27.708013v1?rss=1">
<title>
<![CDATA[
Blood-brain barrier dysfunction predicts cognitive trajectory after ischemic stroke 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.27.708013v1?rss=1</link>
<description><![CDATA[
Ischemic stroke doubles the risk of dementia.1-4 Stroke severity and location affect cognition early,5,6 but late dementia risk is not related to infarct characteristics, nor is it reduced by preventing additional strokes,3,6,7 and its mechanism is unknown. We identified a plasma proteomic signature of chronic stroke that was consistent with blood-brain barrier (BBB) dysfunction, including a 58% decrease in plasma levels of platelet-derived growth factor B and downregulation of its pathway compared to healthy controls. During 2 years of follow-up, the stroke-specific proteome was accentuated in stroke survivors who subsequently declined in the processing speed/executive function cognitive domain. To test BBB function, we performed dynamic contrast-enhanced MRI 6-9 months after stroke in an additional cohort and found 1.7-fold higher whole brain BBB leakage compared to controls. Finally, we compared autopsy tissue from people with infarcts and dementia at death to those with infarcts and no dementia. Those who died with dementia had dramatic loss of vascular mural cell coverage compared to those without dementia (median 0.7% vs. 27%). Thus, our proteomic, functional, and structural data implicate chronic BBB dysfunction in cognitive decline late after stroke, revealing potential proteomic and imaging biomarkers and, importantly, a novel target for intervention.
]]></description>
<dc:creator>Xue, L.</dc:creator>
<dc:creator>Jones, O. A.</dc:creator>
<dc:creator>Drag, L.</dc:creator>
<dc:creator>Zera, K. A.</dc:creator>
<dc:creator>Zhu, L.</dc:creator>
<dc:creator>Mlynash, M.</dc:creator>
<dc:creator>Carmichael, N. S.</dc:creator>
<dc:creator>Shu, C.-H.</dc:creator>
<dc:creator>Biesiada, Z.</dc:creator>
<dc:creator>Seong, D.</dc:creator>
<dc:creator>Thomas, O. M.</dc:creator>
<dc:creator>Simmons, E. C.</dc:creator>
<dc:creator>Huang, E.</dc:creator>
<dc:creator>Berry, K.</dc:creator>
<dc:creator>Chung, P.</dc:creator>
<dc:creator>Aslan, A.</dc:creator>
<dc:creator>Xue, R.</dc:creator>
<dc:creator>Rutledge, J. E.</dc:creator>
<dc:creator>Oh, H. S.-H.</dc:creator>
<dc:creator>Yang, L. K.</dc:creator>
<dc:creator>James, T.</dc:creator>
<dc:creator>Ghanem, M.</dc:creator>
<dc:creator>Moran-Losada, P.</dc:creator>
<dc:creator>Poston, K. L.</dc:creator>
<dc:creator>Henderson, V. W.</dc:creator>
<dc:creator>Grainger, J.</dc:creator>
<dc:creator>Allan, S. M.</dc:creator>
<dc:creator>Lansberg, M.</dc:creator>
<dc:creator>Mormino, E. C.</dc:creator>
<dc:creator>Wyss-Coray, T.</dc:creator>
<dc:creator>Kapasi, A.</dc:creator>
<dc:creator>Schneider, J. A.</dc:creator>
<dc:creator>Smith, C. J.</dc:creator>
<dc:creator>Parkes, L. M.</dc:creator>
<dc:creator>Aghaeepour, N.</dc:creator>
<dc:creator>Buckwalter, M. S.</dc:creator>
<dc:date>2026-03-02</dc:date>
<dc:identifier>doi:10.64898/2026.02.27.708013</dc:identifier>
<dc:title><![CDATA[Blood-brain barrier dysfunction predicts cognitive trajectory after ischemic stroke]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.27.708609v1?rss=1">
<title>
<![CDATA[
Reprogramming mRNA localization by targeted RNA-protein interference 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.27.708609v1?rss=1</link>
<description><![CDATA[
RNA binding proteins (RBPs) associate with RNAs in intricate ribonucleoprotein complexes and regulate various aspects of RNA life cycle and, by extension, cell functions. Despite their significance, elucidating the functional contributions of specific RNA-RBP binding events, particularly in long-term phenotypic assays, remains challenging. Here, we harness the specificity of CRISPR/dCas13 to interfere with specific RNA-RBP interactions. We apply this methodology to GA-rich mRNA localization elements which recruit the RNA-binding protein CNBP and serve as platforms for the assembly of mRNA trafficking complexes. We show that dCas13/gRNA binds to target transcripts in a highly specific manner and sterically interferes with CNBP recruitment leading to altered target mRNA localization and cell motility, consistent with the function of the targeted mRNAs. The effectiveness of dCas13/gRNA as a functional interference tool is curtailed by the strength of target mRNA binding as well as by the amount of cytoplasmic gRNA. We describe optimizations and considerations for the stable implementation of this system, to allow the investigation of long-term functional consequences of altered mRNA distributions.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=141 SRC="FIGDIR/small/708609v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Mason, D.</dc:creator>
<dc:creator>Bandyopadhyay, D.</dc:creator>
<dc:creator>Jiwnani, N.</dc:creator>
<dc:creator>Meyer, B.</dc:creator>
<dc:creator>Mili, S.</dc:creator>
<dc:date>2026-03-02</dc:date>
<dc:identifier>doi:10.64898/2026.02.27.708609</dc:identifier>
<dc:title><![CDATA[Reprogramming mRNA localization by targeted RNA-protein interference]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.28.708767v1?rss=1">
<title>
<![CDATA[
Structural variants in human congenital heart disease disrupt distal genomic regulatory contacts of developmental genes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.28.708767v1?rss=1</link>
<description><![CDATA[
Predicting the functional significance of structural variants (SVs) associated with genetic diseases remains challenging. To test the hypothesis that SVs from people with congenital heart disease (CHD) disrupt developmental chromatin interactions, we developed CardioAkita, a machine-learning model that predicts how variants alter 3D chromatin structure. Analyzing previously genotyped de novo SVs (dnSVs), we observed a positive association between CHD severity and CardioAkita scores across dozens of families. From whole-genome sequencing of three individuals with CHD we predicted disruptive dnSVs. Induced pluripotent stem cells engineered to harbor these variants confirmed CardioAkitas predictions of 3D chromatin changes, and further revealed aberrant expression of local genes including cardiac developmental genes, suggesting that chromatin reorganization plays a significant mechanistic role in the genetic etiology of CHD. Our findings highlight the potential for models of 3D chromatin organization to predict the pathogenicity and underlying mechanisms of SVs in human disease.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Wu, J.</dc:creator>
<dc:creator>Pittman, M.</dc:creator>
<dc:creator>Grant, Z.</dc:creator>
<dc:creator>Kuang, S.</dc:creator>
<dc:creator>Quait, D.</dc:creator>
<dc:creator>Morton, S.</dc:creator>
<dc:creator>Fudenberg, G.</dc:creator>
<dc:creator>Traglia, M.</dc:creator>
<dc:creator>Hayes, K.</dc:creator>
<dc:creator>Pediatric Cardiac Genomics Consortium,</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Bruneau, B.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:date>2026-03-02</dc:date>
<dc:identifier>doi:10.64898/2026.02.28.708767</dc:identifier>
<dc:title><![CDATA[Structural variants in human congenital heart disease disrupt distal genomic regulatory contacts of developmental genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.28.708706v1?rss=1">
<title>
<![CDATA[
NMDA receptor-dependent presynaptic homeostatic plasticity? 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.28.708706v1?rss=1</link>
<description><![CDATA[
Excitatory glutamatergic synapses in the brain are remarkably plastic. Two forms of plasticity have received the most attention: long-term potentiation (LTP) and synaptic homeostasis. While LTP requires the activation of NMDA receptors, synaptic homeostasis does not. However, both phenomena are mediated by the recruitment of postsynaptic AMPA receptors to the synapses. Recently a new form of plasticity has been described referred to as presynaptic homeostatic plasticity (PHP) (Chipman et al., 2022; Chipman et al., 2025). Pharmacological inhibition of AMPA synaptic responses in CA1 hippocampal pyramidal cells initiates a rapid homeostatic response that results in the recovery of the AMPA responses to normal values in the continued presence of the inhibitor. Accompanying this recovery is a doubling of the NMDA response which is interpreted as an increase in the release of glutamate. This is provocative since it is the first report claiming that a reduction in AMPA responses triggers an enhancement in NMDA responses. Using three different protocols to monitor synaptic responses we fail to observe any recovery of synaptic responses in the presence of an AMPA inhibitor. Furthermore, there was no enhancement in NMDA responses. Thus, we find no evidence for the presence of PHP at CA1 hippocampal synapses.
]]></description>
<dc:creator>dou, t.</dc:creator>
<dc:creator>zhang, j.</dc:creator>
<dc:creator>hong, y.</dc:creator>
<dc:creator>chen, x.</dc:creator>
<dc:creator>nicoll, r.</dc:creator>
<dc:date>2026-03-03</dc:date>
<dc:identifier>doi:10.64898/2026.02.28.708706</dc:identifier>
<dc:title><![CDATA[NMDA receptor-dependent presynaptic homeostatic plasticity?]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.01.708910v1?rss=1">
<title>
<![CDATA[
Intestinal intraepithelial lymphocyte migration and tissue homeostasis are dictated by Gα13 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.01.708910v1?rss=1</link>
<description><![CDATA[
The intestinal barrier harbors numerous intraepithelial lymphocytes (IEL) that promote tissue integrity and whose dysfunction contributes to disease. The signals regulating IEL dynamics are incompletely understood. Here we show that deficiency in heterotrimeric G-protein subunit G13 or effector Arhgef1 causes a loss of CD8 and CD8{beta} IEL. Following G13 ablation, IEL reduce migration speed prior to undergoing cell death. Induction of CD8{beta} IEL by Listeria monocytogenes is intact but the cells fail to distribute along the villi and quickly die. TGF{beta} gain-of-function rescues Arhgef1-deficient CD8{beta} cell numbers. CRISPR screening identifies a role for G13-coupled GPR132 in IEL homeostasis. T cell G13-deficient mice suffer more severe colitis and colorectal tumor growth. In summary, we identify G13 as an essential signaling node in {beta} and {gamma}{delta} CD8 IEL and we propose G13-guided IEL positioning in the villous niche is required for receipt of signals, including TGF{beta}, for their maturation, survival and function.
]]></description>
<dc:creator>Earley, Z. M.</dc:creator>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Knopper, K.</dc:creator>
<dc:creator>Peng, F.</dc:creator>
<dc:creator>Qiu, L.</dc:creator>
<dc:creator>Jo, N.</dc:creator>
<dc:creator>Lisicka, W.</dc:creator>
<dc:creator>Taglinao, H.</dc:creator>
<dc:creator>An, J.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Yang, L. V.</dc:creator>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Looney, M. R.</dc:creator>
<dc:creator>Cyster, J. G.</dc:creator>
<dc:date>2026-03-03</dc:date>
<dc:identifier>doi:10.64898/2026.03.01.708910</dc:identifier>
<dc:title><![CDATA[Intestinal intraepithelial lymphocyte migration and tissue homeostasis are dictated by Gα13]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.02.708135v1?rss=1">
<title>
<![CDATA[
NINTEDANIB AND PIRFENIDONE AFFECT GROWTH AND DIFFERENTIATION OF HUMAN ALVEOLAR TYPE 2 CELLS 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.02.708135v1?rss=1</link>
<description><![CDATA[
BackgroundIdiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disease characterized by epithelial cell senescence. Pirfenidone and nintedanib are approved drugs for the treatment of IPF. They significantly slow disease progression, but their mechanisms of action, especially on alveolar type 2 (AT2) cells, are poorly understood. We addressed this question by evaluating colony formation and growth of human AT2 cells co-cultured with fibroblasts in organoid culture in the presence of pirfenidone and nintedanib. We further evaluated molecular changes induced by these drugs via single cell RNA-seq of treated organoids.

MethodsAT2 cells isolated from normal donor lungs or IPF patients were mixed with human fibroblasts in 3D culture and grown in the absence or presence of pirfenidone or nintedanib. After 14 days in culture, the organoids were quantified and cells extracted from Matrigel for single cell RNA-seq.

ResultsAT2 cell organoids cultured in the presence of pirfenidone or nintedanib resulted in increased colony formation and, in the case of nintedanib, in larger colonies. We observed that untreated or pirfenidone treated AT2 cells lost surfactant protein C (SFTPC) expression and acquired an expression profile consistent with keratin (KRT)17high/KRT5- basaloid cells, whereas a larger proportion of nintedanib treated cells retained SFTPC expression. In contrast, AT2 cells treated with TGF{beta} inhibitor exhibited intermediate (SFTPC-/KRT17low) gene expression profile.

ConclusionThese results suggest that nintedanib maintains an AT2-like expression state in culture and acts proximal to TGF{beta}.

Conflict of Interest StatementPJW was supported by grants from Boehringer Ingelheim, Roche, Sanofi, Pliant and Arda Therapeutics and received personal fees from Boehringer Ingelheim and Sanofi. None of these companies had a role in the design or analysis of the study or in the writing of the manuscript. ASM, GP, JRR and DG are employees of Genetech. The other authors have no conflicts of interest to declare.
]]></description>
<dc:creator>Bazarov, A. V.</dc:creator>
<dc:creator>Serra-Marques, A.</dc:creator>
<dc:creator>Protti, G.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Naikawadi, R. P.</dc:creator>
<dc:creator>Green, G.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Kukreja, J.</dc:creator>
<dc:creator>Matthay, M.</dc:creator>
<dc:creator>Wax, M.</dc:creator>
<dc:creator>Cai, X.</dc:creator>
<dc:creator>Wolters, R.</dc:creator>
<dc:creator>Rock, J. R.</dc:creator>
<dc:creator>Garfield, D.</dc:creator>
<dc:creator>Wolters, P. J.</dc:creator>
<dc:date>2026-03-03</dc:date>
<dc:identifier>doi:10.64898/2026.03.02.708135</dc:identifier>
<dc:title><![CDATA[NINTEDANIB AND PIRFENIDONE AFFECT GROWTH AND DIFFERENTIATION OF HUMAN ALVEOLAR TYPE 2 CELLS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.02.709108v1?rss=1">
<title>
<![CDATA[
Hormone signaling and immune programs define differential endocrine responsiveness in high-risk breast tissue 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.02.709108v1?rss=1</link>
<description><![CDATA[
Hormone therapies are frequently used to reduce breast cancer risk in individuals at increased risk for primary or subsequent disease; however, tissue-level responses to these therapies are heterogeneous and incompletely understood. Background parenchymal enhancement (BPE) on breast magnetic resonance imaging (MRI) provides a non-invasive radiologic readout of breast tissue features associated with endocrine responsiveness and cancer risk. Although BPE is associated with hormonal exposure, a subset of patients with BPE do not show a response to preventive endocrine therapy and therefore may remain at increased breast cancer risk. In this study, we integrated single-nucleus RNA sequencing and spatial transcriptomics to define the determinants of endocrine responsiveness in the setting of BPE. We identify hormone-driven epithelial cells with high levels of estrogen signaling and endocrine responsiveness, together with immune-associated epithelial programs characterized by diminished luminal identity and increased expression of immune-modulatory pathways, including major histocompatibility complex (MHC) class II and CD74. Functional organoid assays validate that these epithelial states exhibit differential sensitivity to tamoxifen and demonstrate that inflammatory signals can induce immune-modulatory epithelial programs. Together, our findings identify hormone signaling and immune programs as key determinants of endocrine responsiveness in breast tissue and provide a biological basis for interpreting radiologic markers relevant to cancer prevention.
]]></description>
<dc:creator>Goldhammer, N.</dc:creator>
<dc:creator>Bont, M.</dc:creator>
<dc:creator>Warhadpande, S.</dc:creator>
<dc:creator>Choi, M.</dc:creator>
<dc:creator>Cedano, J.</dc:creator>
<dc:creator>Greenwood, H.</dc:creator>
<dc:creator>Ye, J.</dc:creator>
<dc:creator>Schwartz, C.</dc:creator>
<dc:creator>Alvarado, M.</dc:creator>
<dc:creator>Ewing, C.</dc:creator>
<dc:creator>Goodwin, K.</dc:creator>
<dc:creator>Mukhtar, R.</dc:creator>
<dc:creator>Wong, J.</dc:creator>
<dc:creator>Abe, S.</dc:creator>
<dc:creator>Chandler, J.</dc:creator>
<dc:creator>Jackson, J.</dc:creator>
<dc:creator>Olopade, O.</dc:creator>
<dc:creator>Campbell, M.</dc:creator>
<dc:creator>Lam, A.</dc:creator>
<dc:creator>Park, C.</dc:creator>
<dc:creator>Vertido, A.</dc:creator>
<dc:creator>van 't Veer, L.</dc:creator>
<dc:creator>Hylton, N.</dc:creator>
<dc:creator>Esserman, L.</dc:creator>
<dc:creator>Rosenbluth, J.</dc:creator>
<dc:date>2026-03-04</dc:date>
<dc:identifier>doi:10.64898/2026.03.02.709108</dc:identifier>
<dc:title><![CDATA[Hormone signaling and immune programs define differential endocrine responsiveness in high-risk breast tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.03.709397v1?rss=1">
<title>
<![CDATA[
Myofibroblast lineage mapping and inhibiting subretinal fibrosis by targeting SMAD3 and MRTF pathways via microRNA-24 functional study 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.03.709397v1?rss=1</link>
<description><![CDATA[
Subretinal fibrosis underlies the end-stage pathogenesis of retinal diseases including age- related macular degeneration (AMD). It can disrupt retinal structure and eventually lead to legal blindness by generating contractile force, fibrotic scarring, subretinal hemorrhage, and retinal detachment. Myofibroblasts are the predominant cells critically involved in subretinal fibrosis, however, the cellular contribution to myofibroblasts remains unclear. Here we demonstrate that multiple cell lineages, including macrophages, endothelial cells (EC), retinal pigment epithelial (RPE) cells and pericytes, significantly contribute to myofibroblasts in a laser-induced subretinal fibrosis model. We found microRNA miR-24 is significantly downregulated in the plasma of wet AMD patients. Overexpression of miR-24 represses epithelial-mesenchymal transition (EMT), endothelial-mesenchymal transition (EndMT), and the resulting fibrosis by regulating TGF- {beta}/SMAD3 and PAK4/LIMK2/MRTF pathways. Consistently, a combination of SMAD3 and MRTF inhibitors show superior efficacy to individual inhibitors in repressing fibrosis in vitro and laser-induced subretinal fibrosis in vivo. Together, these suggest the contribution of multiple cell-types in myofibroblast transformation in subretinal fibrosis, and repression of miR-24-regulated TGF-{beta}/SMAD3 and PAK4/LIMK2/MRTF pathways in multiple cell types holds therapeutic potential for treating subretinal fibrosis in AMD and other fibrotic disorders.
]]></description>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Tong, Y.</dc:creator>
<dc:creator>Byrnes, K. G.</dc:creator>
<dc:creator>Zhou, Q.</dc:creator>
<dc:creator>Dong, C.</dc:creator>
<dc:creator>Benjamin, C.</dc:creator>
<dc:creator>Parker, E.</dc:creator>
<dc:creator>Bao, D.</dc:creator>
<dc:creator>Ren, Z.</dc:creator>
<dc:creator>Anderson, C. A.</dc:creator>
<dc:creator>Ufret-Vincenty, R. L.</dc:creator>
<dc:creator>He, Y.-G.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Hinkle, D.</dc:creator>
<dc:creator>Ma, J.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:date>2026-03-04</dc:date>
<dc:identifier>doi:10.64898/2026.03.03.709397</dc:identifier>
<dc:title><![CDATA[Myofibroblast lineage mapping and inhibiting subretinal fibrosis by targeting SMAD3 and MRTF pathways via microRNA-24 functional study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.02.709138v1?rss=1">
<title>
<![CDATA[
LSD Relaxes Structural Constraints on Brain Dynamics and Default Mode Decoupling Tracks Ego Dissolution 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.02.709138v1?rss=1</link>
<description><![CDATA[
Psychedelics profoundly alter conscious experience, yet how they reshape the relationship between brain anatomy and function remains unclear. In particular, it is unknown whether psychedelic states reflect a global disruption of structure-function organization or a frequency- and network-specific reconfiguration of neural dynamics relative to the structural connectome. Here we address this question using source-localized magnetoencephalography mapped onto connectome harmonics to quantify structure-function coupling in humans under lysergic acid diethylamide (LSD) and placebo. LSD induces a robust decoupling of low-frequency (theta, alpha and beta) activity from anatomical constraints, indicating a global loosening of structure-aligned large-scale dynamics. In contrast, high-frequency gamma activity shows selective reorganization rather than uniform disruption. Greater gamma-band decoupling within core default-mode network regions predicts the intensity of ego dissolution across individuals, demonstrating that while LSD broadly alters large-scale dynamics, subjective loss of self may be specifically linked to frequency-selective reorganization of the default-mode network. Functional decoding reveals that LSD does not produce indiscriminate disintegration but instead drives system-specific rebalancing, with preferential decoupling of visual and attentional systems and strengthened coupling within auditory networks. Together, these findings provide electrophysiological evidence that psychedelic states emerge from a frequency-dependent relaxation of structural constraints on brain activity and identify default-mode reorganization as a neural correlate of ego dissolution. These results offer a mechanistic framework for understanding how LSD may exert therapeutic effects by transiently relaxing rigid structural constraints and enhancing dynamical flexibility within networks involved in self-related processing.
]]></description>
<dc:creator>Subramani, V.</dc:creator>
<dc:creator>Pascarella, A.</dc:creator>
<dc:creator>Brunel, J.</dc:creator>
<dc:creator>Harel, Y.</dc:creator>
<dc:creator>Muthukumaraswamy, S. D.</dc:creator>
<dc:creator>Carhart-Harris, R.</dc:creator>
<dc:creator>Jerbi, K.</dc:creator>
<dc:creator>Lioi, G.</dc:creator>
<dc:creator>Farrugia, N.</dc:creator>
<dc:date>2026-03-05</dc:date>
<dc:identifier>doi:10.64898/2026.03.02.709138</dc:identifier>
<dc:title><![CDATA[LSD Relaxes Structural Constraints on Brain Dynamics and Default Mode Decoupling Tracks Ego Dissolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.05.709942v1?rss=1">
<title>
<![CDATA[
ENS lineage potential is not intrinsically regionalized but is modulated by PTPRZ1 signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.05.709942v1?rss=1</link>
<description><![CDATA[
The enteric nervous system (ENS) orchestrates critical gastrointestinal functions including peristalsis, fluid exchange, and blood flow regulation, and develops from vagal neural crest (vNC) progenitors that colonize the gut. While the gut epithelium and mesenchyme exhibit pronounced anterior-posterior (A-P) transcriptional patterning and lineage diversification after mid-gestation, whether the ENS itself undergoes comparable regional embryonic transcriptional diversification has remained unclear. Here, we use multiplexed single-cell RNA sequencing and functional perturbations to dissect how the ENS is patterned between E13.5 and E18.5 within the context of a regionally specialized gut. We find that, while the epithelium and mesenchyme display strong and enduring AP-graded gene expression programs, the ENS lacks intrinsic regionalization and instead follows a predominantly temporal maturation trajectory characterized by neuronal and glial differentiation states. Integrative ligand-receptor analyses reveal that mesenchymal populations express A-P patterned microenvironmental cues that correlate with subtle, region-linked transcriptional tuning in ENS cells, despite the absence of intrinsic A-P identities. Among these signals, PTN/MDK-PTPRZ1 signaling emerges as a major spatial and temporal input to the ENS, with gradients that track both small intestinal region and developmental time. To test the relevance of PTPRZ1 signaling for human ENS development, we perturbed pluripotent stem cell-derived ENS cultures and found that modulating PTPRZ1 signaling impacts proliferative, neurogenic, and neurotransmitter-specification programs, confirming that niche-derived cues fine-tune ENS development. Together, our findings support a model in which the small intestine establishes A-P regionalization through epithelial and mesenchymal patterning, whereas the ENS maintains a relatively uniform core neuroglial program that is secondarily refined by localized microenvironmental signals. This framework highlights how extrinsic, region-specific cues, rather than intrinsic regional transcriptional codes, shape ENS maturation within the small intestine.
]]></description>
<dc:creator>Kalantari, A.</dc:creator>
<dc:creator>Klein, O.</dc:creator>
<dc:creator>Gartner, Z. J.</dc:creator>
<dc:creator>Fattahi, F.</dc:creator>
<dc:date>2026-03-07</dc:date>
<dc:identifier>doi:10.64898/2026.03.05.709942</dc:identifier>
<dc:title><![CDATA[ENS lineage potential is not intrinsically regionalized but is modulated by PTPRZ1 signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.05.708917v1?rss=1">
<title>
<![CDATA[
CEACAM5/6+ Tumor Cells and IL-1β+ Macrophages Drive Resistance to Chemo-immunotherapy in Gastric Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.05.708917v1?rss=1</link>
<description><![CDATA[
Chemo-immunotherapy is a first-line treatment for advanced gastric cancer, yet response rates remain limited and resistance mechanisms are poorly defined. Here we generate a single-cell atlas of 542,121 cells from 35 patients treated with anti-PD-1 plus chemotherapy, profiling pre- and post-treatment tumors linked to clinical response. Integrating spatial transcriptomics, immunohistochemistry, and bulk RNA sequencing, we identify two temporally distinct resistance programs. Intrinsic resistance in pre-treatment non-responders is marked by enrichment of CEACAM5/6 tumor cells that form immune-excluded spatial niches characterized by macrophage recruitment and CD8 T-cell exhaustion. Acquired resistance in post-treatment non-responders is driven by expansion of IL-1{beta} macrophages, which induces coordinated NF-{kappa}B activation across tumor and stromal compartments, promoting PD-L1 upregulation, epithelial-mesenchymal transition, and chronic inflammation. These findings delineate an evolutionary trajectory of resistance and nominate CEACAM5/6 and IL-1{beta} as predictive biomarkers and therapeutic targets to improve anti-PD-1-based combination strategies.
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Han, X.</dc:creator>
<dc:creator>Kang, M.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Xun, Z.</dc:creator>
<dc:creator>Chen, G.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Xu, S.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Hao, Z.</dc:creator>
<dc:creator>Han, Y.</dc:creator>
<dc:creator>Lin, Q.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Liang, H.</dc:creator>
<dc:date>2026-03-08</dc:date>
<dc:identifier>doi:10.64898/2026.03.05.708917</dc:identifier>
<dc:title><![CDATA[CEACAM5/6+ Tumor Cells and IL-1β+ Macrophages Drive Resistance to Chemo-immunotherapy in Gastric Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.07.710284v1?rss=1">
<title>
<![CDATA[
A population-scale red blood cell proteome reveals genetically encoded aging clocks predictive of hemolysis and blood donor activity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.07.710284v1?rss=1</link>
<description><![CDATA[
As the most abundant human cell and the foundation of transfusion medicine, red blood cells (RBCs) offer a unique readout of systemic health, yet they have never been characterized at population scale. We generated a proteome atlas of 13,091 blood donors with multi-omics longitudinal phenotyping, characterizing the influence of demographics and genetic variation on the reproducibility of RBC proteomes across donations. Elastic-net aging clocks captured biological aging with high accuracy and uncovered genetic regulators of {Delta}Age at FN1, C4/IKZF1, CRAT, PFAS, TRIM58. Across independent cohorts, {Delta}Age was accelerated in G6PD deficiency, sickle cell trait/disease, and iron deficiency, reversed by iron repletion, and slowed in high-frequency donors, linking molecular aging to brain iron/myelin and cognitive performance. Molecular aging signatures predicted storage, osmotic, and oxidative hemolysis, hemoglobin increments after transfusion, and long-term donor activity over 12-years. These results establish RBC proteomics as a scalable biomarker of aging, donor healthspan, and transfusion outcomes.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/710284v1_ufig1.gif" ALT="Figure 1">
View larger version (96K):
org.highwire.dtl.DTLVardef@15bd46eorg.highwire.dtl.DTLVardef@1d7c107org.highwire.dtl.DTLVardef@1c1d870org.highwire.dtl.DTLVardef@168dc6f_HPS_FORMAT_FIGEXP  M_FIG Dzieciatkowska et al. generate the first population-scale atlas of the RBC proteome across 13,000 donors and develop proteomic and metabolomic aging clocks that quantify biological age. Molecular {Delta}Age is reproducible across donations, genetically encoded and accelerated in G6PD deficiency, sickle cell trait/disease, and iron deficiency - yet reset by iron repletion, tracking with cognitive function and brain iron/myelin. RBC aging clocks predict hemolytic fragility, transfusion efficacy, and donor activity 12 years later.

C_FIG HighlightsO_LIRBC proteome atlas of 13,091 donors reveals demographic and genetic programs
C_LIO_LIGenetically encoded RBC aging clocks identify regulators of molecular {Delta}age
C_LIO_LIMolecular aging features predict hemolysis and transfusion response across cohorts
C_LIO_LIRBC molecular age forecasts long-term donor activity over a 12-year follow-up
C_LI
]]></description>
<dc:creator>Dzieciatkowska, M.</dc:creator>
<dc:creator>Issaian, A. V.</dc:creator>
<dc:creator>Keele, G. R.</dc:creator>
<dc:creator>Saviola, A.</dc:creator>
<dc:creator>Stephenson, D.</dc:creator>
<dc:creator>Bevers, S.</dc:creator>
<dc:creator>Reisz, J. A.</dc:creator>
<dc:creator>Haiman, Z. B.</dc:creator>
<dc:creator>Nemkov, T.</dc:creator>
<dc:creator>Fang, F.</dc:creator>
<dc:creator>Moore, A.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Kleinman, S.</dc:creator>
<dc:creator>Norris, P. J.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Thein, S.-L.</dc:creator>
<dc:creator>Hod, E. A.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>Roubinian, N. H.</dc:creator>
<dc:creator>Page, G. P.</dc:creator>
<dc:creator>Hansen, K.</dc:creator>
<dc:creator>D'Alessandro, A.</dc:creator>
<dc:date>2026-03-08</dc:date>
<dc:identifier>doi:10.64898/2026.03.07.710284</dc:identifier>
<dc:title><![CDATA[A population-scale red blood cell proteome reveals genetically encoded aging clocks predictive of hemolysis and blood donor activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.07.710099v1?rss=1">
<title>
<![CDATA[
The React & Rebound Model: Capturing Emotion Regulation Dynamics from Passive Wearable Data 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.07.710099v1?rss=1</link>
<description><![CDATA[
BackgroundEmotion regulation--the ability to respond to and restore equilibrium after emotional perturbations--is central to mental health. Yet objective measurement remains limited to lab-based studies with group-level results, while consumer wearables focus on physical activity-related metrics rather than emotional dynamics.

ObjectiveWe aimed to develop computational models that extract personalized, interpretable emotion regulation parameters from continuous heart rate variability (HRV) data collected via consumer wearables during everyday life, and validate these parameters against self-reported anxiety symptoms.

MethodsWe analyzed 4 weeks of continuous HRV data from N = 49 healthy adults wearing Samsung Galaxy Active 2 smartwatches. We derived a continuous autonomic balance signal and developed three computational modeling approaches of increasing sophistication: (1) a static sympathetic load metric, (2) an Ornstein-Uhlenbeck (OU) dynamical systems model capturing continuous restoration dynamics, and (3) a discrete-state Markov transition model--the React & Rebound model-- capturing reactivity and rebound dynamics. All models were estimated using joint hierarchical Bayesian models that simultaneously extract subject-specific parameters from HRV time series and estimate their association with Generalized Anxiety Disorder 7-item scale (GAD-7) scores. The validity of extracted parameters was evaluated against anxiety symptom severity.

ResultsStatic sympathetic load correlated modestly with GAD-7 (r = 0.39, R2 = 0.16). The OU model captured 69% of variance (R2 = 0.69), and the React & Rebound model captured 60% (R2 = 0.60) with substantially fewer parameters. Both models revealed that anxiety symptom severity is associated with the interaction between activation and restoration parameters--not either alone. Fast rebound appeared protective even for highly reactive individuals, who scored comparably to low-reactivity groups when restoration was rapid (Cohens d = 1.17 between highest- and lowest-risk quadrants). In the OU model, the interaction effect was specific to GAD-7 scores versus PHQ-9 and ISI scores; in the React & Rebound model, the interaction was credible across all three symptom measures. Both models were unchanged after controlling for physical activity ({Delta}R2 < 0.002).

ConclusionsComputational models can extract interpretable emotion regulation parameters from naturalistic wearable data. The React & Rebound model yields two personalized parameters--reactivity and rebound--that are strongly associated with anxiety symptoms and define meaningful autonomic profiles. These parameters bridge autonomic dynamics measurable via consumer devices to neural circuit models of emotion regulation, with implications for characterizing individual autonomic profiles via consumer wearables.
]]></description>
<dc:creator>Heusser, A. C.</dc:creator>
<dc:creator>Simon, T. J.</dc:creator>
<dc:creator>Elliot, E.</dc:creator>
<dc:creator>James, C.</dc:creator>
<dc:creator>Gazzaley, A.</dc:creator>
<dc:creator>Gibson, N.</dc:creator>
<dc:date>2026-03-10</dc:date>
<dc:identifier>doi:10.64898/2026.03.07.710099</dc:identifier>
<dc:title><![CDATA[The React & Rebound Model: Capturing Emotion Regulation Dynamics from Passive Wearable Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.09.710667v1?rss=1">
<title>
<![CDATA[
A Dimeric Rocaglate Promotes Multivalent eIF4A-RNA Assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.09.710667v1?rss=1</link>
<description><![CDATA[
Ligand dimerization represents a powerful strategy to enhance avidity, potency, and selectivity. Leveraging the natural-product molecular glue Rocaglamide (RocA), we identified BisRoc, a dimeric rocaglate ligand that potently and durably suppresses translation and exhibits greater specificity across a cancer cell line panel than the monomeric RocA. CRISPRi screening revealed that BisRoc activity is influenced by cellular context, including IFITM-mediated uptake, ABC-type efflux transporters, and the translation initiation factor eIF4A2. Mechanistic studies showed that the paralogs eIF4A1 and eIF4A2 are differentially sensitive to BisRoc-induced dimerization. Owing to the presence of multiple binding sites on RNAs, BisRoc-bridged eIF4A-RNA motifs assemble into higher-order complexes that promote stress-granule formation more efficiently than monomeric RocA. Given the widespread multivalency of RNA-RBP interactions, this ligand dimerization strategy may be extended to modulate the higher-order assembly of other RNA-binding proteins.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=95 SRC="FIGDIR/small/710667v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Shokat, K.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Moore, M. K.</dc:creator>
<dc:creator>Lou, K.</dc:creator>
<dc:creator>Wassarman, D. R.</dc:creator>
<dc:creator>Arab, A.</dc:creator>
<dc:creator>Ojeda, S.</dc:creator>
<dc:creator>Karakyriakou, B.</dc:creator>
<dc:creator>Koglin, A.-S.</dc:creator>
<dc:creator>Ott, C. J.</dc:creator>
<dc:creator>Gilbert, L.</dc:creator>
<dc:date>2026-03-10</dc:date>
<dc:identifier>doi:10.64898/2026.03.09.710667</dc:identifier>
<dc:title><![CDATA[A Dimeric Rocaglate Promotes Multivalent eIF4A-RNA Assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.10.710584v1?rss=1">
<title>
<![CDATA[
Metabolic control of drug resistance by a mycobacterial ion channel 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.10.710584v1?rss=1</link>
<description><![CDATA[
Pyrazinamide (PZA) is a cornerstone of modern tuberculosis therapy, yet its context-dependent activity has obscured both its mode of action and resistance mechanisms. Using a host-mimicking culture system integrated with genome-wide CRISPRi profiling, metabolomics, and comparative genomics, we identify a previously unrecognized driver of PZA resistance in humans: loss of the ion channel Rv2571c. Rv2571c mediates -ketoglutarate efflux, amplifying PZA-induced cytoplasmic acidification under host-relevant acidic conditions. Loss-of-function mutations confer resistance in vitro and in vivo and are under positive selection in clinical isolates, establishing this pathway as a resistance determinant in patients. Together, these findings define a novel, ion channel-mediated resistance mechanism, establish cytoplasmic acidification as the basis of PZA killing, and inform resistance detection and treatment-shortening drug development.
]]></description>
<dc:creator>Gouzy, A.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Na, A.</dc:creator>
<dc:creator>Saleh, A.</dc:creator>
<dc:creator>Azadian, Z. A.</dc:creator>
<dc:creator>Tam, K.</dc:creator>
<dc:creator>Munsamy-Govender, V.</dc:creator>
<dc:creator>Poulton, N. C.</dc:creator>
<dc:creator>DeJesus, M. A.</dc:creator>
<dc:creator>Schnappinger, D.</dc:creator>
<dc:creator>Rhee, K. Y.</dc:creator>
<dc:creator>Ehrt, S.</dc:creator>
<dc:creator>Rock, J. M.</dc:creator>
<dc:date>2026-03-10</dc:date>
<dc:identifier>doi:10.64898/2026.03.10.710584</dc:identifier>
<dc:title><![CDATA[Metabolic control of drug resistance by a mycobacterial ion channel]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.09.710372v1?rss=1">
<title>
<![CDATA[
Genome-scale mapping of variant, enhancer and gene function in primary human CD4+ T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.09.710372v1?rss=1</link>
<description><![CDATA[
CD4+ T cells harbor a disproportionate enrichment of immune disease risk loci and represent the primary cellular context for immune disease biology, yet the genes and regulatory programs these variants affect remain largely unknown. We combined targeted Perturb-seq of 1,032 cis-regulatory elements (CREs) overlapping 4,724 variants across 14 immune diseases with genome-wide Perturb-seq of all expressed genes in primary human CD4+ T cells, spanning 4.1 million cells. We identified 626 CRE-gene pairs, and connected CRE targets to downstream regulatory cascades. At the TYK2 and DEXI/CLEC16A loci, we resolved target genes and linked noncoding variants to inflammatory and metabolic programs. Across diseases, we revealed that dispersed variants converged on shared and disease-specific programs. Our work provides a framework for tracing variant-to-CRE-to-gene-to-network in disease-relevant primary cells.
]]></description>
<dc:creator>Moonen, D. P.</dc:creator>
<dc:creator>Claringbould, A.</dc:creator>
<dc:creator>Gschwind, A. R.</dc:creator>
<dc:creator>Schrod, S.</dc:creator>
<dc:creator>Braunger, J.</dc:creator>
<dc:creator>Feng, C.</dc:creator>
<dc:creator>Rauscher, B.</dc:creator>
<dc:creator>Yi, J.</dc:creator>
<dc:creator>Bi, S. Z.</dc:creator>
<dc:creator>Matthess, Y.</dc:creator>
<dc:creator>Kaulich, M.</dc:creator>
<dc:creator>Acob, R. A.</dc:creator>
<dc:creator>Ayer, A.</dc:creator>
<dc:creator>Engreitz, J. M.</dc:creator>
<dc:creator>Velten, B.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Trynka, G.</dc:creator>
<dc:creator>Zaugg, J. B.</dc:creator>
<dc:creator>Schraivogel, D.</dc:creator>
<dc:creator>Steinmetz, L. M.</dc:creator>
<dc:date>2026-03-11</dc:date>
<dc:identifier>doi:10.64898/2026.03.09.710372</dc:identifier>
<dc:title><![CDATA[Genome-scale mapping of variant, enhancer and gene function in primary human CD4+ T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.10.710694v1?rss=1">
<title>
<![CDATA[
KMT2C and KMT2D amplify GRHL2-driven enhancer activation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.10.710694v1?rss=1</link>
<description><![CDATA[
The activation of cis-regulatory enhancers is essential for cell fate specification by driving cell type-specific gene expression. Differentiation models are widely used to study enhancer biology but the asynchronous and interdependent nature of gene regulatory changes during cell state transitions can complicate mechanistic studies. To overcome these limitations, here we develop a tamoxifen-gated system for acute enhancer activation in embryonic stem cells (ESCs) based on GRHL2, a pioneer transcription factor which naturally becomes expressed as naive ESCs differentiate into the formative ESC state. Using this system, we investigate the functional relationship between GRHL2 and the histone mono-methyltransferases KMT2C and KMT2D (KMT2C/D). GRHL2 readily binds its target sites independent of KMT2C/D. However, in the absence of KMT2C/D, there are dramatic reductions in H3K4me1/2, P300 recruitment, and H3K27ac deposition at these sites as well as diminished transcriptional activation. Still, strikingly, a basal level of active enhancer mark acquisition and transcriptional activation occurs. Consistent with these findings, during the naive to formative ESC differentiation, GRHL2 enhancer remodeling and target expression is also strongly but incompletely dependent on KMT2C/D. Together, these results define a functional co-activator relationship in which KMT2C/D act as important amplifiers of GRHL2-driven enhancer activation in ESCs and establish a rapid inducible system for dissecting the kinetics and enzymatic dependencies of pioneer transcription factor mediated enhancer remodeling.
]]></description>
<dc:creator>Boileau, R. M.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Blelloch, R.</dc:creator>
<dc:date>2026-03-11</dc:date>
<dc:identifier>doi:10.64898/2026.03.10.710694</dc:identifier>
<dc:title><![CDATA[KMT2C and KMT2D amplify GRHL2-driven enhancer activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.10.710854v1?rss=1">
<title>
<![CDATA[
Saxiphilin is a broad-spectrum toxin sponge for C13-modified saxitoxins 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.10.710854v1?rss=1</link>
<description><![CDATA[
Saxitoxin (STX) and its congeners (paralytic shellfish toxins, PSTs) are among the most potent small-molecule toxins. PSTs are produced by harmful algal blooms and derive toxicity by disrupting voltage-gated sodium channel (NaV) bioelectrical signaling. Understanding how PST structural variation affects target binding is crucial to develop means to counteract PSTs and exploit these natural products as drug development leads. Frog and toad saxiphilins (Sxphs) are soluble, high-affinity STX toxin sponge proteins that offer a powerful platform to define PST-protein interactions. Here, we show that American bullfrog (Rana catesbeiana) RcSxph and High Himalaya frog (Nanorana parkeri) NpSxph bind a broad set of C13-modified STX congeners. High-resolution X-ray crystal structures of toxin complexes with RcSxph, RcSxph mutants, and NpSxph unveil two C13-aryl congener binding modes, termed  compact and  open, that depend on the RcSxph Tyr558 local environment. These results highlight a remarkable adaptability of Sxphs for accommodating chemically diverse STX analogs and reveal unexpected toxin conformational plasticity. These findings have implications for understanding PST interactions with biological targets and informing design of STX-based probes and NaV modulators.
]]></description>
<dc:creator>Zakrzewska, S.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Park, E.</dc:creator>
<dc:creator>Bhaskar, R. G.</dc:creator>
<dc:creator>Bedell, T. A.</dc:creator>
<dc:creator>Du Bois, J.</dc:creator>
<dc:creator>Minor, D. L.</dc:creator>
<dc:date>2026-03-11</dc:date>
<dc:identifier>doi:10.64898/2026.03.10.710854</dc:identifier>
<dc:title><![CDATA[Saxiphilin is a broad-spectrum toxin sponge for C13-modified saxitoxins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.09.710658v1?rss=1">
<title>
<![CDATA[
Interferon-α and -β subtypes have temporally distinct roles in containing viral spread and protecting vital organs. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.09.710658v1?rss=1</link>
<description><![CDATA[
Type I interferons (IFN-Is), including a single IFN-{beta} and over a dozen IFN-s, induce the anti-viral state. In vitro, IFN-{beta} transcription requires the assembly of the "enhanceosome" composed of the constitutive transcription factors NF-{kappa}B and AP-1, and either constitutive IRF3 or IFN-I-inducible IRF7. IRF3 and IRF7 transcribe mouse IFN-4 and human IFN-1/13. Only IRF7 transcribes other IFN-s. How IFN-I subtype multiplicity and their differential constitutive/IFN-I-inducible versus only IFN-I-inducible transcription help control viruses in vivo remains unknown. Using novel genetically modified mice, we demonstrate that most or all IFN-I subtypes, regardless of their transcriptional control, are necessary to curb the systemic dissemination of lymph-borne ectromelia virus (ECTV) but do not necessarily suppress ECTV or West Nile Virus (WNV) replication in the liver or the brain, or promote survival to their infection. Individually, the most critical IFN-I subtype to survive ECTV and WNV infections is IFN-{beta}. IFN-4 potentiates IFN-{beta} but is not essential. PRDII, the IFN-{beta} promoters NF-{kappa}B binding site, which is required for enhanceosome assembly, is dispensable for in vivo IFN-{beta} production but complements IRF7-IFN-{beta} transcription to restrain ECTV and WNV. IFN-{beta} alone protects IFN--deficient mice from ECTV but not from WNV lethality. Control of WNV replication in the brain successively requires IFN-, IFN-{beta}, and then PRDII-dependent IFN-{beta}, suggesting that brain protection requires IFN-I production by various cell types and pathways. Thus, contrary to the prevailing view, in vivo, IFN-{beta} does not function earlier than IFN-, and IFN-I subtypes non-redundantly cooperate to restrain viruses in the periphery and in vital organs.
]]></description>
<dc:creator>Rezende Melo da Silva, C.</dc:creator>
<dc:creator>Roman, M. I.</dc:creator>
<dc:creator>Heath, N.</dc:creator>
<dc:creator>Gelman, M.</dc:creator>
<dc:creator>Keller, M.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Tang, L.</dc:creator>
<dc:creator>Taiwo, O.</dc:creator>
<dc:creator>Kafle, S.</dc:creator>
<dc:creator>Torres-Gonzalez, E.</dc:creator>
<dc:creator>Ramage, H.</dc:creator>
<dc:creator>Andino, R.</dc:creator>
<dc:creator>Sigal, L. J.</dc:creator>
<dc:date>2026-03-12</dc:date>
<dc:identifier>doi:10.64898/2026.03.09.710658</dc:identifier>
<dc:title><![CDATA[Interferon-α and -β subtypes have temporally distinct roles in containing viral spread and protecting vital organs.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.09.710682v1?rss=1">
<title>
<![CDATA[
Hypothalamic Orexin Input to the Medial Amygdala Links Vigilance to Arousal 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.09.710682v1?rss=1</link>
<description><![CDATA[
Arousal circuits govern anesthetic state transitions, but emergence is often complicated by agitation, and the neural circuits linking anesthetic arousal to vigilance remain unclear. Here, we identify a lateral hypothalamic orexin-to-medial amygdala pathway (LHAOx[-&gt;]MeA) that links anesthetic state transitions to vigilance-like behavioral bias. Synaptic labeling and slice recordings revealed dense LHAOx innervation in the MeA, and RNAscope showed predominant Ox2R expression in MeA GABAergic (MeAvGAT) neurons. LHAOx[-&gt;]MeA terminal activation increased local orexin release, promoted arousal from isoflurane anesthesia, delayed loss of righting reflex during induction, and accelerated recovery during emergence. In vigilance assays, stimulation acutely suppressed open-field exploration and produced real-time place preference without conditioned reinforcement. Fiber photometry demonstrated preferential recruitment of MeAvGAT neurons across anesthetic states. Direct MeAvGAT activation recapitulated cortical engagement, arousal, and vigilance phenotypes, whereas TeLC-mediated silencing of MeAvGAT output abolished induction delay and reversed vigilance-like bias while sparing light-anesthesia arousal and emergence acceleration.
]]></description>
<dc:creator>Xiang, X.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:date>2026-03-12</dc:date>
<dc:identifier>doi:10.64898/2026.03.09.710682</dc:identifier>
<dc:title><![CDATA[Hypothalamic Orexin Input to the Medial Amygdala Links Vigilance to Arousal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.11.711234v1?rss=1">
<title>
<![CDATA[
BMP antagonism is required for mandible outgrowth in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.11.711234v1?rss=1</link>
<description><![CDATA[
The Meckels cartilage (MC) is a fundamental component of mandibular development across vertebrates. In mammals, MC is transient and functions primarily as an early template for mandibular ossification, whereas other vertebrates, including zebrafish, retain MC within the mandible throughout life. Despite its importance, the requirements for MC in sustaining mandibular growth and how signaling pathways implicated in MC development contribute to this process remain unclear. Here, we investigated the dosage-dependent roles of BMP antagonists during zebrafish MC development using mutant alleles of grem1a, nog2, and nog3. Compound mutant adults exhibited fully penetrant mandibular truncation. MC shortening emerged after early larval stages, indicating a requirement for BMP antagonism to sustain cartilage growth. Chondrocyte number remained unchanged as phenotypes developed, but mutants displayed disorganized cartilage morphology and increased chondrocyte volume. Molecular analyses revealed reduced col2a1a domains and expanded ihha and col10a1a expression, consistent with ectopic hypertrophic-like differentiation. Constitutive activation of BMP receptor signaling in chondrocytes recapitulated these phenotypes. Although osteogenesis appeared unaffected by 14 dpf, loss of a tnn skeletal mesenchyme population was observed. Together, these findings demonstrate that BMP antagonists sustain MC growth by regulating chondrocyte differentiation and cartilage organization to support mandibular growth in non-mammalian vertebrates.

Summary StatementThis study leverages zebrafish to define the cellular and molecular mechanisms by which BMP antagonism sustains mandibular growth.
]]></description>
<dc:creator>Chen, H.-J.</dc:creator>
<dc:creator>Dukov, J.</dc:creator>
<dc:creator>Llyod, T.</dc:creator>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Farmer, D.</dc:creator>
<dc:date>2026-03-12</dc:date>
<dc:identifier>doi:10.64898/2026.03.11.711234</dc:identifier>
<dc:title><![CDATA[BMP antagonism is required for mandible outgrowth in zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.11.711097v1?rss=1">
<title>
<![CDATA[
GAP mimetic activity of pan-Ras TCI daraxonrasib synergizes with K-Ras Switch-II pocket inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.11.711097v1?rss=1</link>
<description><![CDATA[
Tricomplex inhibitors (TCIs) are a novel class of direct Ras inhibitors that target the GTP-bound Ras(on) state trough recruitment of Cyclophilin A. Daraxonrasib (RMC-6236) is a pan-Ras TCI that was recently shown to restore GTPase activity of G12-mutant Ras proteins. Structural analysis of a pan-Ras TCI bound to K-Ras(GDP- AlF3) reveals a transition-state arrangement of Tyr32 and Gln61 that closely resembles endogenous GTPase- GAP complexes. This includes a closed Switch-I conformation engaging the cis-GTPase machinery in a manner analogous to non-arginine-finger GAPs such as RanGAP. These observations position pan-Ras TCIs as pharmacologic GAP mimetics. The GTPase-promoting activity of daraxonrasib suggests synergy with Switch-II pocket K-Ras inhibitors, including the approved GDP-state selective K-Ras G12C inhibitor adagrasib (MRTX-849), whose engagement of K-Ras(GTP) is kinetically constrained by slow endogenous hydrolysis of the mutant GTPase. We demonstrate that daraxonrasib sensitizes K-Ras(GTP) to adagrasib labeling in both recombinant protein and cellular context. In K-Ras G12C and G12D mutant cell lines, combinations of daraxonrasib with adagrasib or HRS-4642 (MRTX-1133 analog) yield more rapid K-Ras engagement, rapid p-ERK suppression, and significant Loewe synergy scores in viability assays. These findings establish GAP mimetics as rational and potent combination partners for SW-II inhibitors. The synergistic combination has potential to deepen and prolong pathway suppression while enabling dose reductions that may mitigate on-target toxicity and resistance.
]]></description>
<dc:creator>Pfaff, P.</dc:creator>
<dc:creator>Shokat, K.</dc:creator>
<dc:date>2026-03-13</dc:date>
<dc:identifier>doi:10.64898/2026.03.11.711097</dc:identifier>
<dc:title><![CDATA[GAP mimetic activity of pan-Ras TCI daraxonrasib synergizes with K-Ras Switch-II pocket inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.12.711465v1?rss=1">
<title>
<![CDATA[
Biophysical trade-offs in antibody evolution are resolved by conformation-mediated epistasis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.12.711465v1?rss=1</link>
<description><![CDATA[
Protein evolution is constrained by multidimensional biophysical factors, in which mutations that enhance one property often compromise another. Antibodies represent an extreme case: they evolve rapidly to bind diverse antigens, yet mutations that improve affinity can disrupt folding, reduce cell-surface trafficking, or promote self-reactivity, and are typically selected against during affinity maturation. Though biophysical characterization of individual antibodies suggests that such trade-offs are pervasive, their impact on antibody evolutionary trajectories remains unclear, in part because existing high-throughput biophysical methods rely on heterologous systems that are often poorly suited for human proteins. Here, we develop a high-throughput platform to quantify multiple biophysical parameters of large libraries of full-length proteins that are natively synthesized, processed, and displayed on human cells. We apply this approach to a human antibody lineage that matures to recognize divergent SARS-CoV-2 variants by measuring the surface expression, antigen affinity, and self-reactivity for all 213 possible evolutionary intermediates between the unmutated and mature sequences. These measurements reveal that mutations differentially affect these biophysical properties - in some cases, improving one property at the expense of another. We leverage these data to compute the likelihood of all possible evolutionary paths, finding that very few paths can navigate these multidimensional requirements. The few accessible paths acquire mutations in a specific order that either circumvent trade-offs between biophysical properties or offset deleterious effects on one property with beneficial effects on another. By determining the structures of the ancestral and evolved antibodies, we find that these coordinated mutational effects arise from a conformational rearrangement that alleviates steric clashes and reshapes the biophysical landscape, enabling otherwise inaccessible mutational paths. Together, this work defines the multidimensional biophysical constraints and structural mechanisms that govern antibody evolution and establishes a general framework for mapping and predicting the biophysical effects of mutations in human proteins.
]]></description>
<dc:creator>Tharp, C. R.</dc:creator>
<dc:creator>Catalano, C.</dc:creator>
<dc:creator>Khalifeh, A.</dc:creator>
<dc:creator>Ghaffari-Kashani, S.</dc:creator>
<dc:creator>Huang, R.</dc:creator>
<dc:creator>Kang, G.</dc:creator>
<dc:creator>Scapin, G.</dc:creator>
<dc:creator>Phillips, A. M.</dc:creator>
<dc:date>2026-03-13</dc:date>
<dc:identifier>doi:10.64898/2026.03.12.711465</dc:identifier>
<dc:title><![CDATA[Biophysical trade-offs in antibody evolution are resolved by conformation-mediated epistasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.12.711375v1?rss=1">
<title>
<![CDATA[
Robust PHP in Adult Hippocampus: Essential Assay Optimizations 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.12.711375v1?rss=1</link>
<description><![CDATA[
Presynaptic homeostatic plasticity (PHP) is a potent form of homeostatic plasticity that has been documented at synapses as diverse as the glutamatergic Drosophila neuromuscular junction (NMJ), cholinergic mammalian NMJ (including human), and glutamatergic synapses in the mammalian brain. Published experimental evidence in favor of PHP in adult hippocampus and cerebellum includes patch-clamp electrophysiology, presynaptic capacitance measurement, calcium imaging, optical reporters of vesicle release and correlated three-dimensional electron microscopy. These studies are grounded in newly optimized experimental protocols that differ substantively from those typically used to study activity-dependent plasticity in neonatal and juvenile slice preparations. Here, we elaborate and extend our assays and methodologies for the study of PHP in the adult mammalian brain. Our assays are designed to optimize synapse, cell and tissue health and minimize the incorporation of unintended adverse experimental conditions that may interfere with the induction and/or expression of PHP. In addition, we provide benchmark criteria for assessment of cell health, necessary for analysis of PHP and, in so doing, advance our understanding of postsynaptic conditions necessary for PHP induction in the adult brain. Our data underscore why PHP may have been previously overlooked, inclusive of a recent manuscript challenging the robust expression of PHP in the mammalian brain (Dou et al., 2026 BioRxiv [preprint]).
]]></description>
<dc:creator>Chipman, P. H.</dc:creator>
<dc:creator>Fetter, R. D.</dc:creator>
<dc:creator>Ragozzino, F. J.</dc:creator>
<dc:creator>Lee, U.</dc:creator>
<dc:creator>Davis, G. W.</dc:creator>
<dc:date>2026-03-16</dc:date>
<dc:identifier>doi:10.64898/2026.03.12.711375</dc:identifier>
<dc:title><![CDATA[Robust PHP in Adult Hippocampus: Essential Assay Optimizations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.12.711398v1?rss=1">
<title>
<![CDATA[
Reinforcement Learning for Antibiotic Stewardship: Optimizing Prescribing Policies Under Antimicrobial Resistance Dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.12.711398v1?rss=1</link>
<description><![CDATA[
Antimicrobial resistance (AMR) threatens antibiotic effectiveness, but quantitatively evaluating stewardship strategies under partial observability and delayed feedback remains difficult in real-world data. We developed abx_amr_simulator, a Gymnasium-compatible simulation framework, and used it to benchmark reinforcement learning (RL) prescribing policies against value-iteration (VI) benchmarks and fixed prescribing rules across four experiment sets of increasing complexity, with varying levels of information degradation.

Across scenarios, temporal abstraction was consistently important: flat PPO was competitive only in simpler settings, whereas hierarchical PPO was generally needed when prescribing decisions had delayed, coupled effects on future resistance. We found that adding recurrent memory did not uniformly improve performance; its value was context-dependent. In some degraded-information settings, memoryless policies performed better by adopting conservative update-responsive behavior, while in more complex, multi-signal partially observable settings, recurrent memory provided modest advantages.

Patient heterogeneity and risk-stratification signals were major determinants of policy quality. When agents could differentiate higher-from lower-risk patients, they more reliably learned selective treatment behavior, stabilized AMR, and improved clinical outcomes. Exaggerated risk stratification modestly outperformed accurate stratification, while compressed stratification produced moderate degradation. In more realistic settings combining noisy patient observations, delayed AMR surveillance, and multi-patient decisions, hierarchical agents outperformed fixed prescribing rules across both stewardship and clinical metrics, converging to conservative low-AMR equilibria with reduced cross-seed variance.

Across experiments, results support the utility of hierarchical RL as a best-case policy-analysis tool for stewardship under uncertainty, while also highlighting that performance estimates are sensitive to observation structure and training horizon design. The framework provides a controlled environment for hypothesis generation and for stress-testing prescribing strategies before translation to policy-relevant settings.
]]></description>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Blumberg, S.</dc:creator>
<dc:date>2026-03-16</dc:date>
<dc:identifier>doi:10.64898/2026.03.12.711398</dc:identifier>
<dc:title><![CDATA[Reinforcement Learning for Antibiotic Stewardship: Optimizing Prescribing Policies Under Antimicrobial Resistance Dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.13.711689v1?rss=1">
<title>
<![CDATA[
Survey of the human proteostasis network: the ubiquitin-proteasome system 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.13.711689v1?rss=1</link>
<description><![CDATA[
Modification by ubiquitination governs the half-lives of thousands of proteins that are fated for elimination by either the proteasome or autophagy pathways, depending on the intricate architectures of ubiquitin modification. This system mediates quality control for individual proteins, protein complexes, and organelles, as well as myriad purely regulatory functions. Here we provide a comprehensive survey of the ubiquitin-proteasome system (UPS), the scope of which is at present poorly defined. The UPS, with the inclusion of pathways involving ubiquitin-like modifiers, comprises in our estimate over 1400 distinct proteins in humans, a vast set of activities whose collective impact on the biology of the cell is pervasive. The UPS is an integral component of the proteostasis network (PN), the remainder of which we have also surveyed in recent studies. With the addition of molecular chaperones, proteins from autophagy-lysosome pathway, and related activities, the PN includes in total over 3100 components by our estimates. Comprehensive and systematic definition of these pathways should support a range of ongoing investigations in the areas of genomics, proteomics, biochemistry, cell biology, and disease research.
]]></description>
<dc:creator>Elsasser, S.</dc:creator>
<dc:creator>Powers, E.</dc:creator>
<dc:creator>Stoeger, T.</dc:creator>
<dc:creator>Sui, X.</dc:creator>
<dc:creator>Kurtzbard, R. D.</dc:creator>
<dc:creator>Martinez-Botia, P.</dc:creator>
<dc:creator>Wangaline, M. A.</dc:creator>
<dc:creator>Gama, A. R.</dc:creator>
<dc:creator>Huttlin, E. L.</dc:creator>
<dc:creator>Elia, L. P.</dc:creator>
<dc:creator>Kelly, J. W.</dc:creator>
<dc:creator>Gestwicki, J. E.</dc:creator>
<dc:creator>Frydman, J. E.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:creator>Clerico, E. M.</dc:creator>
<dc:creator>Morimoto, R.</dc:creator>
<dc:creator>Prado, M. A.</dc:creator>
<dc:creator>Vertegaal, A. C. O.</dc:creator>
<dc:creator>Hofmann, K.</dc:creator>
<dc:creator>Finley, D.</dc:creator>
<dc:date>2026-03-16</dc:date>
<dc:identifier>doi:10.64898/2026.03.13.711689</dc:identifier>
<dc:title><![CDATA[Survey of the human proteostasis network: the ubiquitin-proteasome system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.13.711624v1?rss=1">
<title>
<![CDATA[
Single-library chromosome-scale diploid assemblies of vole genomes resolve a species-specific duplication implicated in pair bonding 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.13.711624v1?rss=1</link>
<description><![CDATA[
High-quality reference genomes are essential to effectively characterize genomic drivers of speciation, phenotypic diversity, and disease causality. Larger complex genomes often require integration of long-read DNA sequencing with additional genomic data, such as chromosome conformation capture (Hi-C or CiFi) to generate phased chromosome-scale assemblies, however this requires multiple sequencing platforms (in the case of Hi-C) or the construction of multiple long-read sequencing libraries. Here, we devise a strategy that combines PacBio HiFi and CiFi sequencing in a single library and run to efficiently produce high-quality contiguous chromosome-scale diploid genome assemblies. We apply this approach to liver tissue from single individuals of prairie vole (Microtus ochrogaster) and meadow vole (Microtus pennsylvanicus), generating haplotype-resolved, chromosome-scale 2.3 Gbp genomes with QV~62, and 99.3% BUSCO completeness. Comparing the two new genomes identifies complex structural changes impacting Avpr1a, previously implicated in pair bonding, including a species-specific duplication missing from the existing prairie vole reference genome. These divergent genomic features offer new avenues of investigation related to behavioral divergence between prairie and meadow voles. This single-library approach facilitates a simplified and more affordable assembly workflow, producing near-complete genomes of diverse species using one sequencing platform.
]]></description>
<dc:creator>Abuelanin, M.</dc:creator>
<dc:creator>Kaya, G.</dc:creator>
<dc:creator>Lake, J. A.</dc:creator>
<dc:creator>Lambert, C.</dc:creator>
<dc:creator>Wu, M. V.</dc:creator>
<dc:creator>Berendzen, K.</dc:creator>
<dc:creator>Krasheninnikova, K.</dc:creator>
<dc:creator>Wood, J. M.</dc:creator>
<dc:creator>Solomon, N. G.</dc:creator>
<dc:creator>Donaldson, Z. R.</dc:creator>
<dc:creator>Bales, K. L.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Korlach, J.</dc:creator>
<dc:creator>Manoli, D. S.</dc:creator>
<dc:creator>Tollkuhn, J.</dc:creator>
<dc:creator>Dennis, M. Y.</dc:creator>
<dc:date>2026-03-17</dc:date>
<dc:identifier>doi:10.64898/2026.03.13.711624</dc:identifier>
<dc:title><![CDATA[Single-library chromosome-scale diploid assemblies of vole genomes resolve a species-specific duplication implicated in pair bonding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.16.712192v1?rss=1">
<title>
<![CDATA[
Using image classifiers to predict CMT2A disease-relevant mitochondrial motility phenotypes in iPSC motor neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.16.712192v1?rss=1</link>
<description><![CDATA[
Charcot-Marie-Tooth disease type 2A (CMT2A) is a genetic disease characterized by autosomal dominant MFN2 mutations and dysregulated mitochondrial trafficking. While there is currently no FDA-approved CMT2A therapy, the recent development of iPSC motor neuron model systems, high-throughput imaging platforms, and CRISPR-based gene editing technologies holds promise for screening new therapies at scale in vitro. A critical roadblock for therapeutic screening is the development of scalable and robust computational methods to assess the mitochondrial trafficking phenotypes, healthy or diseased, of each iPSC motor neuron sample. To address this gap, we developed a vision transformer (ViT) based classification framework that predicts disease phenotypes using kymographs, an image transformation that captures particle movement along a prespecified path, such as mitochondrial movement along axons. We show that our classification approach more accurately discriminates healthy MFN2 wild-type (WT) from diseased MFN2 R364W-mutant (R364W) iPSCs than alternative summary statistics, such as mitochondrial speed and fraction of stationary mitochondria that are directly extracted from kymographs. Furthermore, we show that our model maintains high accuracy when deployed on a biological replicate holdout dataset. An analysis of ViT patch embeddings of the kymographs shows that mitochondria with highly variable sizes and many intersection events most strongly associate with R364W diseased kymographs. The computational approach demonstrated in this paper has broad applicability for future high-throughput screens where organelle trafficking along axons plays a key role in disease pathogenesis.
]]></description>
<dc:creator>Epstein, L.</dc:creator>
<dc:creator>Weiner, A. C.</dc:creator>
<dc:creator>Macklin, B.</dc:creator>
<dc:creator>Kelly, K. R.</dc:creator>
<dc:creator>Conklin, B. R.</dc:creator>
<dc:creator>Engelhardt, B. E.</dc:creator>
<dc:date>2026-03-17</dc:date>
<dc:identifier>doi:10.64898/2026.03.16.712192</dc:identifier>
<dc:title><![CDATA[Using image classifiers to predict CMT2A disease-relevant mitochondrial motility phenotypes in iPSC motor neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.16.712262v1?rss=1">
<title>
<![CDATA[
Preferential formation of NUP98-KDM5A condensates at specific H3K4me3-rich loci drives leukemogenic gene expression 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.16.712262v1?rss=1</link>
<description><![CDATA[
Chromosomal translocations involving NUP98 generate fusion proteins that alter gene expression programs, yet the fundamental principles governing their gene targeting and condensate behavior remain poorly understood. Using NUP98-KDM5A as a model, we integrate cellular imaging, in vitro reconstitution, and genomic analyses to dissect how chromatin engagement shapes condensate formation. We find that NUP98-KDM5A forms sub-diffraction-limited, gel-like condensates whose assembly is potentiated by binding to H3K4me3. This interaction creates a quantitative targeting mechanism in which, at the native expression level, condensates preferentially form at genomic loci with high local H3K4me3 density. Such local density-dependent recruitment explains selective enrichment at the leukemogenic HOX gene clusters, despite widespread presence of H3K4me3 across the genome. Analysis of single-cell sequencing data from patients further supports a correlation between local H3K4me3 density and transcriptional activation in NUP98-KDM5A-driven leukemia. Together, our findings reveal how activating chromatin marks and condensate-forming proteins synergize to generate specificity within euchromatin, offering a generalizable framework for understanding how chromatin-associated condensates interpret epigenetic landscapes.
]]></description>
<dc:creator>Berrocal, A.</dc:creator>
<dc:creator>Sandoval, J. E.</dc:creator>
<dc:creator>Khetan, N.</dc:creator>
<dc:creator>Ma, A.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Narlikar, G. J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Galonic Fujimori, D.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:date>2026-03-17</dc:date>
<dc:identifier>doi:10.64898/2026.03.16.712262</dc:identifier>
<dc:title><![CDATA[Preferential formation of NUP98-KDM5A condensates at specific H3K4me3-rich loci drives leukemogenic gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.16.711480v1?rss=1">
<title>
<![CDATA[
Genome-scale functional mapping of the mammalian whole brain with in vivo Perturb-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.16.711480v1?rss=1</link>
<description><![CDATA[
Functional genomics studies have provided critical insights into cell type-specific gene regulatory programs, but to date most have been conducted in wild-type tissues or cell cultures. Here, we present a gene expression functional atlas across the mouse brain. We use an enhanced in vivo Perturb-seq platform to analyze transcriptome-wide responses to loss of 1,947 disease-associated genes, profiling over 7.7 million cells spanning major brain regions and neuronal populations. We find striking cell-type-specific essentiality and transcriptional programs and show that closely related disease genes such as two NMDA receptor subunits can drive opposing transcriptional programs. Together, this work reveals insights into the genetics and mechanisms of neurodevelopmental, psychiatric, and neurodegenerative diseases in vivo, paving the way for the design of future genetic medicine.
]]></description>
<dc:creator>Shi, T.</dc:creator>
<dc:creator>Korshunova, M.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>DeTomaso, D.</dc:creator>
<dc:creator>Zheng, X.</dc:creator>
<dc:creator>Vishvanath, L.</dc:creator>
<dc:creator>Nyasulu, T.</dc:creator>
<dc:creator>Huynh, N.</dc:creator>
<dc:creator>Sun, A.</dc:creator>
<dc:creator>Thompson, P. C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Wigdor, E. M.</dc:creator>
<dc:creator>Rohani, N.</dc:creator>
<dc:creator>Ali, S.</dc:creator>
<dc:creator>Qiu, H.</dc:creator>
<dc:creator>Geralt, M.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Rabhi, S.</dc:creator>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>van Velthoven, C. T.</dc:creator>
<dc:creator>Nery, J. R.</dc:creator>
<dc:creator>Castanon, R. G.</dc:creator>
<dc:creator>Dicks, S.</dc:creator>
<dc:creator>Chen, T. J.</dc:creator>
<dc:creator>Ecker, J. R.</dc:creator>
<dc:creator>Zeng, H.</dc:creator>
<dc:creator>Zheng, G. X.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Sundaram, L.</dc:creator>
<dc:creator>Jin, X.</dc:creator>
<dc:date>2026-03-18</dc:date>
<dc:identifier>doi:10.64898/2026.03.16.711480</dc:identifier>
<dc:title><![CDATA[Genome-scale functional mapping of the mammalian whole brain with in vivo Perturb-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.16.711194v1?rss=1">
<title>
<![CDATA[
Female-enriched Eggerthella lenta drives neuroinflammation and IFN-γ via host receptor TLR2 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.16.711194v1?rss=1</link>
<description><![CDATA[
Women are at increased risk of autoimmune diseases, including multiple sclerosis (MS); however, the degree to which sex differences in the gut microbiota impact autoimmunity remains largely unexplored. Our 27-cohort meta-analysis revealed 60 sex-associated gut bacterial species. Leveraging an independent clinical cohort, we demonstrate that female-enriched species significantly associate with MS status and clinical disability (EDSS). Top female-enriched species Eggerthella lenta drove disease in the experimental autoimmune encephalomyelitis (EAE) MS model, consistent with brain and gut lamina propria T cell infiltration and MS-associated T helper (Th) signatures. E. lenta induced intestinal Th1 and Th17 in healthy mice, independent of bacterial viability. Mechanistically, we demonstrate that TLR2 directly drives E. lenta-induced IFN-{gamma} production in Th cells and is necessary for exacerbation of EAE. Together, we identify a causal host-microbe axis contributing to sex differences in autoimmunity and provide a framework for evaluating sex as a biological variable in human microbiome research.

HIGHLIGHTSO_LI27-cohort meta-analysis identifies a robust sex-signature in human gut microbiota.
C_LIO_LIFemale-enriched species are associated with MS risk and severity.
C_LIO_LIFemale-enriched Eggerthella lenta exacerbates the EAE model.
C_LIO_LIE. lenta impacts neuroinflammation via toll-like receptor 2.
C_LI
]]></description>
<dc:creator>Rock, R. R.</dc:creator>
<dc:creator>Alexander, M.</dc:creator>
<dc:creator>Noecker, C.</dc:creator>
<dc:creator>Trepka, K.</dc:creator>
<dc:creator>Upadhyay, V.</dc:creator>
<dc:creator>Ortega, E.</dc:creator>
<dc:creator>Ramirez, L.</dc:creator>
<dc:creator>Siewart, L.</dc:creator>
<dc:creator>Olson, C.</dc:creator>
<dc:creator>Halsey, T.</dc:creator>
<dc:creator>Probstel, A.-K.</dc:creator>
<dc:creator>Baranzini, S.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:date>2026-03-19</dc:date>
<dc:identifier>doi:10.64898/2026.03.16.711194</dc:identifier>
<dc:title><![CDATA[Female-enriched Eggerthella lenta drives neuroinflammation and IFN-γ via host receptor TLR2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.16.711909v1?rss=1">
<title>
<![CDATA[
A Three-dimensional Analytical Framework for Retinal Microvasculature Reveals Layer-associated Vulnerability in Development and Neovascular Remodeling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.16.711909v1?rss=1</link>
<description><![CDATA[
The neurosensory retina is one of the most metabolically active tissues in the body and a uniquely accessible extension of the central nervous system, where neuronal and vascular structures can be visualized non-invasively. Its accessibility and highly organized laminar architecture make it a powerful model for studying vascular development and a window into systemic health. Although computational analyses of retinal images have enabled risk assessment for ocular and systemic diseases, most vascular studies rely on two-dimensional frameworks with limited resolution of capillary structure and layer-specific organization. Here, we present a high-resolution three-dimensional (3D) imaging and analysis pipeline enabling quantification of retinal microvasculature and extraction of structural and network metrics across vascular layers. We apply this approach to two mouse models of aberrant retinal vascular development: one with spontaneous postnatal chorioretinal neovascularization and another with disrupted neurovascular lattice formation and layered organization in early life. Across both pathologic contexts, 3D analysis enables detailed characterization of retinal vascular architecture and identifies early vulnerability within the intermediate plexus, the vascular network that bridges the superficial and deep retinal layers, as a sensitive indicator of abnormal remodeling and neovascularization. This framework enables precise characterization of retinal vasculature and establishes a foundation for identifying new retinal biomarkers with potential relevance to neurovascular and systemic disease.
]]></description>
<dc:creator>Shang, W.</dc:creator>
<dc:creator>Hong, G.</dc:creator>
<dc:creator>Keller, W. E.</dc:creator>
<dc:creator>Morton, R. A.</dc:creator>
<dc:creator>Zeboulon, P.</dc:creator>
<dc:creator>Kenichi, T.</dc:creator>
<dc:creator>Duan, X.</dc:creator>
<dc:creator>Gould, D. B.</dc:creator>
<dc:creator>Kim, T. N.</dc:creator>
<dc:date>2026-03-19</dc:date>
<dc:identifier>doi:10.64898/2026.03.16.711909</dc:identifier>
<dc:title><![CDATA[A Three-dimensional Analytical Framework for Retinal Microvasculature Reveals Layer-associated Vulnerability in Development and Neovascular Remodeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.18.712472v1?rss=1">
<title>
<![CDATA[
Gene Expansion and Regulatory Rewiring Shape Sex-Biased Evolution of the Mouse Submandibular Gland Secretome 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.18.712472v1?rss=1</link>
<description><![CDATA[
Mammalian saliva plays essential roles in digestion, immunity, and host-microbiome interactions, yet its protein composition varies across species and sexes. The evolutionary mechanisms underlying this molecular diversity remain poorly understood. Here, we compared mouse and human salivary gland secretomes at genomic, transcriptomic, and proteomic levels to understand how saliva composition evolves. We performed RNA-seq analysis of the major mouse salivary glands (parotid, submandibular, and sublingual), liver and pancreas from both sexes, compared them with reanalyzed previously published human salivary gland transcriptomes, and integrated them with proteome data of mouse and human whole saliva. We found that evolution of gene expression in mouse salivary glands is driven by rapid gene turnover and sexual dimorphism. In the submandibular and sublingual glands, respectively, 68% and 73% of expression from genes encoding secreted proteins derives from lineage-specific genes that lack one-to-one human orthologs. Mouse submandibular gland shows striking sexual dimorphism, with 1537 tissue specific sex-biased genes, five times higher than in the liver, a classic model of sex-biased expression. These genes cluster in regions shaped by recent gene duplication, such as the kallikrein gene cluster, a mouse-specific expansion that accounts for [~]16.4% of male-biased submandibular expression. Our analyses suggest that this bias arises through regulatory changes that are expanded by gene duplication, including the spread of a testosterone-associated regulatory motif and the expansion of a shared chromatin domain that promotes coordinated gene regulation. Our results reveal how lineage-specific gene duplication and regulatory rewiring drive rapid, sex-specific evolution of the mammalian salivary gland secretome.
]]></description>
<dc:creator>Landau, L. J. B.</dc:creator>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Griffin, N.</dc:creator>
<dc:creator>Saikia, A.</dc:creator>
<dc:creator>Kramer, J. M.</dc:creator>
<dc:creator>Knox, S.</dc:creator>
<dc:creator>Ruhl, S.</dc:creator>
<dc:creator>Gokcumen, O.</dc:creator>
<dc:date>2026-03-20</dc:date>
<dc:identifier>doi:10.64898/2026.03.18.712472</dc:identifier>
<dc:title><![CDATA[Gene Expansion and Regulatory Rewiring Shape Sex-Biased Evolution of the Mouse Submandibular Gland Secretome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.19.713017v1?rss=1">
<title>
<![CDATA[
Library docking for Cannabinoid-2 Receptor ligands 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.19.713017v1?rss=1</link>
<description><![CDATA[
Cannabinoid receptors are therapeutically promising GPCRs that are also interesting test systems for structure-based methods, which have targeted them previously. Here we used the CB2 receptor as a template to explore several topical questions in library docking. Whereas an earlier campaign against the CB1 receptor led to potent but relatively non-selective ligands, here we found that targeting interactions with polar, orthosteric site residues led to subtype-selective ligands. Docking hit rate and especially hit affinity improved in moving from a 7 million to a 2.6 billion molecule library. Similar to earlier studies, docking against active and inactive states of the receptor did not reliably bias toward the discovery of agonists or inverse agonists. Cryo-EM structures of two of the new agonists, each in a different chemotype, superposed well on the docking predictions. Correspondingly, structure-based optimization led to 10- to 140-fold improvements within three different series, also consistent with well-behaved ligand families. Hit rates with a fully enumerated 2.6 billion molecule library resembled those of an implied 11 billion molecule library from a building-block method, consistent with the latters ability to explore this space, though higher affinities were discovered from the fully enumerated set. Overall, eight diverse families of ligands, with potencies <100 nM and mostly unrelated to previously known ligands were found. Implications for future studies are considered.
]]></description>
<dc:creator>Rachman, M. M.</dc:creator>
<dc:creator>Iliopoulos-Tsoutsouvas, C.</dc:creator>
<dc:creator>Dominic Sacco, M.</dc:creator>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Wu, C.-G.</dc:creator>
<dc:creator>Santos, E.</dc:creator>
<dc:creator>Glenn, I. S.</dc:creator>
<dc:creator>Paris, L.</dc:creator>
<dc:creator>Cahill, M. K.</dc:creator>
<dc:creator>Ganapathy, S.</dc:creator>
<dc:creator>Tummino, T. A.</dc:creator>
<dc:creator>Moroz, Y. S.</dc:creator>
<dc:creator>Radchenko, D. S.</dc:creator>
<dc:creator>Okorie, M.</dc:creator>
<dc:creator>Tawfik, V. L.</dc:creator>
<dc:creator>Irwin, J. J.</dc:creator>
<dc:creator>Makriyannis, A.</dc:creator>
<dc:creator>Skiniotis, G.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:date>2026-03-21</dc:date>
<dc:identifier>doi:10.64898/2026.03.19.713017</dc:identifier>
<dc:title><![CDATA[Library docking for Cannabinoid-2 Receptor ligands]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.19.712907v1?rss=1">
<title>
<![CDATA[
Neurogenin-2 Reprograms Human Microglial Lineage Cells into Neurons In Vitro and in Chimeric Brains 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.19.712907v1?rss=1</link>
<description><![CDATA[
Progressive neuronal loss is a hallmark of many neurological disorders, yet the adult human brain has a limited capacity for endogenous neuronal replacement. Direct neuronal reprogramming represents an alternative strategy for generating new neurons. Microglia, the brains resident immune cells, are uniquely positioned as candidate cellular substrates due to their abundance, self-renewal capacity, high motility, and rapid recruitment to sites of injury. Here, using live-cell imaging and electrophysiological recordings, we show that human pluripotent stem cell (hPSC)-derived primitive macrophage progenitors (PMPs) and their microglial derivatives exhibit neuronal reprogramming competence. Inducible expression of NEUROG2 in hPSC-derived PMPs drives acquisition of neuronal morphology, sequential expression of early and mature neuronal markers, organization of synaptic proteins, and functional excitability characterized by action potential firing. Single-nucleus RNA sequencing reveals a continuous, directionally ordered reprogramming trajectory marked by suppression of myeloid transcriptional programs, progression through intermediate remodeling states, and progressive activation of neuronal gene regulatory networks, consistent with a regulated lineage conversion rather than partial identity switching. Using a xenotransplantation-based human microglia chimeric brain model, we further demonstrate that inducible NEUROG2 expression reprograms donor-derived human microglia toward a neuronal identity in vivo. Together, these findings establish human microglial lineage cells as a previously unexplored substrate for neuronal reprogramming, providing a conceptual framework for microglia-based strategies aimed at neuronal replacement and neural repair.
]]></description>
<dc:creator>Jin, M.</dc:creator>
<dc:creator>Ma, Z.</dc:creator>
<dc:creator>Dang, R.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Xue, H.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Jiang, P.</dc:creator>
<dc:date>2026-03-22</dc:date>
<dc:identifier>doi:10.64898/2026.03.19.712907</dc:identifier>
<dc:title><![CDATA[Neurogenin-2 Reprograms Human Microglial Lineage Cells into Neurons In Vitro and in Chimeric Brains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.22.713509v1?rss=1">
<title>
<![CDATA[
Human ancestors interbred with two distinct populations of distant relatives 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.22.713509v1?rss=1</link>
<description><![CDATA[
Ancient DNA has shown that a distantly-related "superarchaic" population interbred first with the ancestors of Neanderthals and Denisovans and later with Denisovans themselves. Other work has shown that a superarchaic population interbred with the African ancestors of all modern humans. But it is not yet clear whether these events involved the same superarchaic population. Here, we use the distribution of derived alleles among populations to evaluate hypotheses about superarchaics and their relationship to other hominins of the Pleistocene and Holocene. We find evidence for at least two distinct superarchaic populations. The one contributing to archaic Eurasian populations (Denisovans and Neanderthal-Denisovan ancestors) diverged earlier from the human lineage than did the one contributing to early moderns in Africa. These findings reveal previously unrecognized structure among hominin populations of the Pleistocene.
]]></description>
<dc:creator>Rogers, A. R.</dc:creator>
<dc:creator>Islam, M. T.</dc:creator>
<dc:creator>Brand, C. M.</dc:creator>
<dc:creator>Webster, T. H.</dc:creator>
<dc:date>2026-03-23</dc:date>
<dc:identifier>doi:10.64898/2026.03.22.713509</dc:identifier>
<dc:title><![CDATA[Human ancestors interbred with two distinct populations of distant relatives]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.23.713599v1?rss=1">
<title>
<![CDATA[
ZMAP: A single-cell meta-atlas of zebrafish embryonic development reveals a consensus hierarchy of cell identities 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.23.713599v1?rss=1</link>
<description><![CDATA[
Single-cell RNA sequencing (scRNA-seq) efforts have generated large collections of high-resolution cellular atlases of embryonic development, providing unprecedented views of the dynamic gene expression programs that accompany cell fate specification. Alongside parallel efforts in other models, zebrafish has emerged as one of the most extensively profiled vertebrate embryonic systems, with numerous scRNA-seq atlases spanning both embryonic and early larval stages. Despite this progress, cross-study comparisons between datasets remain challenging due to differences in sample processing, mapping, and annotation conventions. Here we present ZMAP (Zebrafish Meta Atlas Project), a harmonized reference integrating 8 published whole-embryo zebrafish scRNA-seq datasets comprising 798,790 cells across 15 developmental time windows. ZMAP unifies component studies through a shared embedding, a standardized marker-gene discovery pipeline, and a hierarchical annotation ontology. Using ZMAP, we inferred "consensus identity programs" - marker gene signatures for each ontology group that were reproducibly detected across studies. To promote broad usage, we provide a Python-based API for automated annotation and retrieval of marker gene sets and reference objects, as well as a web portal that supports interactive 2D and 3D exploration of the UMAP embedding, gene and annotation-level queries, and access to consensus marker resources.
]]></description>
<dc:creator>Aponte-Santiago, N. A.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Wagner, D. E.</dc:creator>
<dc:date>2026-03-24</dc:date>
<dc:identifier>doi:10.64898/2026.03.23.713599</dc:identifier>
<dc:title><![CDATA[ZMAP: A single-cell meta-atlas of zebrafish embryonic development reveals a consensus hierarchy of cell identities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.22.713538v1?rss=1">
<title>
<![CDATA[
Genetic architectures of brain-related traits are shaped by strong selective constraints 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.22.713538v1?rss=1</link>
<description><![CDATA[
Genome-wide association studies (GWAS) have identified hundreds of significant loci for psychiatric disorders, yet the strength of these associations remains modest compared to other human complex traits with similar numbers of hits. Whether this pattern reflects statistical artifacts or real biological differences -- and, if the latter, what underlies it -- remains unclear. In addition to psychiatric disorders, we find that other traits with functional enrichment in the central nervous system (CNS), whether binary or quantitative, also share similar genetic architectures, characterized by GWAS hits of limited statistical significance and generally higher allele frequencies. To robustly compare traits that differ in GWAS statistical power, we demonstrate how binarizing a quantitative trait reduces power. This loss of power can be replicated by a matched "effective sample size" on the liability scale. After matching "effective sample sizes", we show that CNS-enriched traits have large mutational target sizes, with contributing variants and genes experiencing stronger selection than those for other traits. Our findings reveal heterogeneity among diseases and provide insights into traits that more effectively capture fitness-relevant processes. More broadly, our results suggest that the genetic architectures of complex traits are shaped by the tissues through which these traits are mediated.
]]></description>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Simons, Y. B.</dc:creator>
<dc:creator>Spence, J. P.</dc:creator>
<dc:creator>Sella, G.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:date>2026-03-25</dc:date>
<dc:identifier>doi:10.64898/2026.03.22.713538</dc:identifier>
<dc:title><![CDATA[Genetic architectures of brain-related traits are shaped by strong selective constraints]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.23.713595v1?rss=1">
<title>
<![CDATA[
WTR: A Toolkit for Functional Anterograde Transsynaptic Circuit Mapping 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.23.713595v1?rss=1</link>
<description><![CDATA[
The brain coordinates animal physiology and behavior via neuronal circuits. To understand and simulate brain functions, it is essential to delineate the synaptic connectivity between neurons. Transsynaptic tracers serve as powerful tools for such purposes. In response to the demand for anterograde tracers for circuit mapping and functional interrogation, we developed WTR, a fusion protein of mammalian codon-optimized WGA, TEV-protease cleavage sequence, and Recombinase. WTR expressed via AAV vectors in cell-type-specific starter neurons reaches their postsynaptic neurons and releases Cre/Flpo upon exposure to TEV-protease expressed in downstream neurons. Accompanied by Cre/Flpo-dependent expression of EGFP, GCaMP7s, or ChR2, the toolkit enables labeling, recording, or manipulation of downstream neurons. We utilized WTR to characterize downstream neurons of either glutamatergic or GABAergic neurons in the preoptic area of anterior hypothalamus for their differential actions in thermoregulation or stress responses, respectively. These results establish WTR as a versatile platform for functional anterograde circuit mapping.
]]></description>
<dc:creator>Wang, T. A.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Liu, R.</dc:creator>
<dc:creator>Yi-Luo, A.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Hu, J.</dc:creator>
<dc:creator>Guan, S.</dc:creator>
<dc:creator>Chang, S.-y.</dc:creator>
<dc:creator>Cui, X.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Zhao, F.</dc:creator>
<dc:creator>Huang, C.-T.</dc:creator>
<dc:creator>Duan, X.</dc:creator>
<dc:creator>Jan, L. Y.</dc:creator>
<dc:date>2026-03-25</dc:date>
<dc:identifier>doi:10.64898/2026.03.23.713595</dc:identifier>
<dc:title><![CDATA[WTR: A Toolkit for Functional Anterograde Transsynaptic Circuit Mapping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.23.713717v1?rss=1">
<title>
<![CDATA[
HIV-1 infection does not confer intrinsic resistance to cell death induced by cytotoxic T lymphocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.23.713717v1?rss=1</link>
<description><![CDATA[
To eliminate the persistent reservoir of cells harboring intact HIV-1 proviruses in people living with HIV-1 (PLWH), cure strategies like the Shock-and-Kill approach rely on effector functions of cytolytic T lymphocytes (CTL). CTL are involved in the initial control of HIV-1 viremia and target productively infected cells throughout the course of infection. However, selective killing of susceptible cells could generate a reservoir dominated by cells with dysregulated cell death pathways or other features conferring resistance to killing. Here, we use CTL-engaging single-chain diabodies to assess the rate of lysis of uninfected and HIV-1-infected primary CD4+ T cells under identical CTL pressure in the settings of both latent and active infection. Our findings indicate that with this mode of CTL triggering, infected and uninfected CD4+ T cells from PLWH on ART are generally lysed at identical rates, and that an apparent survival advantage for actively infected CD4+ T cells primarily reflects the reduced surface antigen availability through previously described Nef-dependent downregulation of MHC class I molecules. No survival advantage is observed when the CTL response is directed through diabodies to the stably expressed non-classical MHC class I molecule HLA-E, indicating equal susceptibility to cell death.
]]></description>
<dc:creator>Bachmann, N.</dc:creator>
<dc:creator>Kim, B.</dc:creator>
<dc:creator>Simonetti, F. R.</dc:creator>
<dc:creator>Kovacs, C. M.</dc:creator>
<dc:creator>Hoh, R.</dc:creator>
<dc:creator>Deeks, S. G.</dc:creator>
<dc:creator>Siliciano, J. D.</dc:creator>
<dc:creator>Siliciano, R. F.</dc:creator>
<dc:date>2026-03-25</dc:date>
<dc:identifier>doi:10.64898/2026.03.23.713717</dc:identifier>
<dc:title><![CDATA[HIV-1 infection does not confer intrinsic resistance to cell death induced by cytotoxic T lymphocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.24.714045v1?rss=1">
<title>
<![CDATA[
Heparan sulfate is essential for Drosophila FGF export 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.24.714045v1?rss=1</link>
<description><![CDATA[
Binding of Fibroblast growth factor (FGF) to a heparan sulfate proteoglycan (HSPG) is required for paracrine FGF signaling. To improve our understanding of FGF:HSPG association, we developed a method to monitor export of the Drosophila FGF ortholog Branchless (Bnl) in vivo. We detected Bnl on the surface of approximately 10% of Bnl-producing cells, but Bnl on the surface of cells depleted of HS was much reduced. HS depletion also non-autonomously decreased the activity of cytonemes that extend from cells that receive Bnl. These results are consistent with the idea that Bnl export to the cell surface is regulated, that intracellular binding of an HSPG to Bnl in producing cells is essential for export, and that cells that take up Bnl actively participate in its release from producing cells.

SummaryLevels of FGF exported to the surface of FGF-expressing cells are dependent on intracellular heparan sulfate proteoglycans.
]]></description>
<dc:creator>Barbosa, G. O.</dc:creator>
<dc:creator>Solis-Calero, C.</dc:creator>
<dc:creator>Kornberg, T.</dc:creator>
<dc:date>2026-03-26</dc:date>
<dc:identifier>doi:10.64898/2026.03.24.714045</dc:identifier>
<dc:title><![CDATA[Heparan sulfate is essential for Drosophila FGF export]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.24.712448v1?rss=1">
<title>
<![CDATA[
Requirement for oxidation of neuronal ketone bodies in aging andneurodegeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.24.712448v1?rss=1</link>
<description><![CDATA[
Glucose is the brains primary fuel, but the brain can also use alternative energy substrates, especially during development or starvation. Emerging evidence suggests ketone metabolism may help the brain adapt to energy stress in neurodegenerative diseases such as Alzheimers disease, although its role in constitutive brain function in normal aging is poorly understood. Using iPSC-derived human neurons and adult-inducible, neuron-specific Bdh1 knockout mice, we show that ketone body metabolism is essential for maximum energy production, neuronal function, and mouse survival--even under normal nutritional conditions. Mechanistically, phenotypes of Bdh1 knockout neurons are mitigated by provision of acetoacetate, a downstream energy metabolite. Moreover, loss of neuronal ketone oxidation markedly increases mortality and memory deficits in Alzheimers disease model mice. These findings identify ketones as critical neuronal fuels, with particular importance during neurodegeneration. While non-energetic activities of ketone bodies are increasingly appreciated, oxidation for energy provision is an essential mechanism for normal function in neurons and mice. Targeting the energetic function of ketones may thus offer new therapeutic strategies for both aging and neurodegenerative diseases such as Alzheimers.
]]></description>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Nomura, M.</dc:creator>
<dc:creator>Meng, J. X.</dc:creator>
<dc:creator>Garcia, T. Y.</dc:creator>
<dc:creator>Matsuura, T. R.</dc:creator>
<dc:creator>Kelly, D. P.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Newman, J. C.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.24.712448</dc:identifier>
<dc:title><![CDATA[Requirement for oxidation of neuronal ketone bodies in aging andneurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.24.713239v1?rss=1">
<title>
<![CDATA[
A network for self-transcendence derived from patients with brain lesions 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.24.713239v1?rss=1</link>
<description><![CDATA[
Self-transcendence, the reorientation of experience away from the self toward others, nature, or broader meaning, is a fundamental dimension of human psychology, yet its causal neural architecture remains poorly understood. Here we applied lesion network mapping to 88 neurosurgical patients with pre- and post-operative assessments of trait self-transcendence to identify the distributed brain network whose disruption alters this capacity. The resulting network showed significant spatial correspondence with the default mode network and, at a finer parcellation level, with frontoparietal control subnetworks. Leave-one-out analyses identified posterior midline regions as the most stable correlates of increased self-transcendence following brain lesions. Independent validation against fMRI meta-analyses of self-referential processing, compassion, and ketamine administration, alongside a neuromodulation target previously shown to modulate the sense of self, converged on a consistent model. These findings provide causal evidence for a network architecture in which posterior midline hubs constrain, and brainstem and anterior midline regions facilitate, self-transcendent experience.
]]></description>
<dc:creator>Healey, M. R.</dc:creator>
<dc:creator>Sanchez-Gama, Y.</dc:creator>
<dc:creator>Ding, M.</dc:creator>
<dc:creator>McMahon, J. T.</dc:creator>
<dc:creator>Bourbon, C.</dc:creator>
<dc:creator>Jesani, R.</dc:creator>
<dc:creator>Atwood, G. D.</dc:creator>
<dc:creator>Lord, B. T.</dc:creator>
<dc:creator>Sanguinetti, J.</dc:creator>
<dc:creator>Brewer, J.</dc:creator>
<dc:creator>Vago, D. R.</dc:creator>
<dc:creator>Siddiqi, S. H.</dc:creator>
<dc:creator>Fabbro, F.</dc:creator>
<dc:creator>Urgesi, C.</dc:creator>
<dc:creator>Nielsen, J. A.</dc:creator>
<dc:creator>Ferguson, M. A.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.24.713239</dc:identifier>
<dc:title><![CDATA[A network for self-transcendence derived from patients with brain lesions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.25.713648v1?rss=1">
<title>
<![CDATA[
Mining functional genes and characterizing cellular transcriptomic profiles in the single-cell atlas of adult Spodoptera litura ovary 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.25.713648v1?rss=1</link>
<description><![CDATA[
Understanding the reproductive biology of non-model organisms remains challenging due to the limited availability of high-resolution molecular resources. Here, we present a comprehensive single-cell transcriptomic atlas of the adult ovary of Spodoptera litura (S. litura), a highly polyphagous agricultural pest with a polytrophic meroistuc ovary. By integrating single-cell RNA sequencing with cross-species comparison to Drosophila melanogaster (D. melanogaster), we define major germline and somatic cell populations and delineate conserved and species-specific features of ovarian cell composition. To enhance the interpretability and reuse of this dataset, we combine transcriptomic profiling with in situ hybridization to validate cluster-specific molecular markers across ovarian cell types. We further apply RNA interference (RNAi) to assess the contributions of germline-enriched genes (Hsc70-4, Wech, Polo, Path) to ovarian development and fecundity. Trajectory inference, together with SCENIC and CellChat analyses, provides a system-level view of transcriptional regulatory programs and predicted intercellular communication pathways during oogenesis in S. litura. Collectively, this work establishes a valuable resource for studying lepidopteran insect oogenesis, offering a comparative framework for reproductive biology in non-model insects and highlighting potential targets for RNAi-based pest control strategies.
]]></description>
<dc:creator>Sun, Z.</dc:creator>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Yi, X.</dc:creator>
<dc:creator>Nystul, T. G.</dc:creator>
<dc:creator>Zhong, G.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.25.713648</dc:identifier>
<dc:title><![CDATA[Mining functional genes and characterizing cellular transcriptomic profiles in the single-cell atlas of adult Spodoptera litura ovary]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.25.714309v1?rss=1">
<title>
<![CDATA[
Combinatorial constraints predict that mitochondrial networks contain a large component 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.25.714309v1?rss=1</link>
<description><![CDATA[
Mitochondria often form branching membrane networks distributed throughout the cell interior. In many, though not all, cell types, these networks are observed to consist of one large connected component together with many smaller fragments. Why does this pattern arise? Does it reflect a specific biological function, an external biophysical constraint, or something simpler? Using results from extremal graph theory, we prove a new theorem which suggests that, under a sufficiently broad sampling of the space of mitochondria-like graphs, the predominance of three-way junctions makes the appearance of a large component likely. This suggests that, in some settings, a large component may serve as a useful null model for mitochondrial network structure rather than requiring a dedicated explanation. More broadly, our result points towards testable predictions, since systematic deviations from this baseline may help reveal additional constraints or mechanisms shaping mitochondrial morphology.
]]></description>
<dc:creator>Mostov, R.</dc:creator>
<dc:creator>Lewis, G. R.</dc:creator>
<dc:creator>Das, M.</dc:creator>
<dc:creator>Marshall, W. F.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.25.714309</dc:identifier>
<dc:title><![CDATA[Combinatorial constraints predict that mitochondrial networks contain a large component]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.26.714449v1?rss=1">
<title>
<![CDATA[
Microenvironmental Determinants of Reaction Kinetics in Biomolecular Condensates Probed with Protein Ligation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.26.714449v1?rss=1</link>
<description><![CDATA[
Cells utilize liquid-liquid phase separation to organize biochemical reactions within biomolecular condensates, which function as membraneless organelles. Although these assemblies are known to enhance reaction rates by concentrating reactants, the mechanisms beyond simple mass-action effects remain poorly understood. Here, we examined how the physicochemical microenvironment within condensates modulates reaction kinetics using spontaneous protein ligation as a model reaction, conducting a systematic analysis across various condensates, ranging from structured scaffolds (PRM-SH3 systems) to intrinsically disordered protein (IDP)-based scaffolds such as LAF, TAF, and FUS. We designed a FRET-based proximity-sensitive client probe to quantify increases in effective local concentration arising from excluded-volume effects. In parallel, we measured internal hydrophilicity and water activity, revealing them as additional key determinants of reaction acceleration. Together, the findings presented here elucidate how phase-separated compartments regulate biochemical reactions through the interplay of physical (effective concentration) and chemical (hydrophilicity and water activity) microenvironments and provide mechanistic insights for engineering condensates with tunable reactivity.
]]></description>
<dc:creator>Bae, J.</dc:creator>
<dc:creator>Hong, K.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Jun, J.</dc:creator>
<dc:creator>Jung, Y.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.714449</dc:identifier>
<dc:title><![CDATA[Microenvironmental Determinants of Reaction Kinetics in Biomolecular Condensates Probed with Protein Ligation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.714678v1?rss=1">
<title>
<![CDATA[
Tumor Heterogeneity Induces Pro- and Anti-metastatic Myeloid Cell Phenotypes in Breast Cancer Lung Metastasis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714678v1?rss=1</link>
<description><![CDATA[
Metastasis is a major cause of cancer-related mortality, yet targeting metastatic cells directly has been largely unsuccessful due to their plasticity and heterogeneity. Myeloid cells play diverse pro- and anti-metastatic functions and are an attractive alternative target for treating metastasis, but how tumor heterogeneity influences myeloid cell phenotypes during metastasis remains poorly understood. Here, we profiled myeloid cells from primary tumors and matched metastatic lungs of 12 heterogeneous and differentially metastatic patient-derived xenograft models of breast cancer. Comparative analysis of cell type abundances revealed distinct myeloid remodeling specific to primary tumors or metastatic lungs. Beyond compositional differences, we identified gene expression programs that were associated with metastatic burden, such as number or size of metastatic nodules, indicating distinct microenvironmental requirements for metastatic seeding and outgrowth. Examining these metastasis-associated programs using time-course datasets, we discovered an evolution from anti- to pro-metastatic monocyte phenotypes during metastatic progression. We further showed that this phenotypic shift was driven by an increase in two distinct myeloid-derived suppressor cell signatures, and a transcriptionally regulated impairment of monocyte differentiation leading to the depletion of non-classical monocytes. Our results comprehensively dissect the heterogeneity of myeloid cell phenotypes across primary tumor and metastatic sites, opening novel avenues for myeloid-targeting therapies specific to metastasis.
]]></description>
<dc:creator>Superville, D. A.</dc:creator>
<dc:creator>Chrenkova, E.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Barozzi, I.</dc:creator>
<dc:creator>Werb, Z.</dc:creator>
<dc:creator>Rendeiro, A. F.</dc:creator>
<dc:creator>McGinnis, C.</dc:creator>
<dc:creator>Goga, A.</dc:creator>
<dc:creator>Winkler, J.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714678</dc:identifier>
<dc:title><![CDATA[Tumor Heterogeneity Induces Pro- and Anti-metastatic Myeloid Cell Phenotypes in Breast Cancer Lung Metastasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.714056v1?rss=1">
<title>
<![CDATA[
Metagenomic strain-resolved DNA modification patterns link extrachromosomal genetic elements to host strains 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714056v1?rss=1</link>
<description><![CDATA[
DNA modification is central to microbial defense against extrachromosomal genetic elements (ECEs), consequently ECEs tend to adopt their hosts modification patterns. Shared ECE-host modification patterns enable linking ECEs to their hosts, but modification detection tools are designed for single genomes and are ineffective at metagenome scale. Here, we present MODIFI, software for detecting DNA modifications in metagenomes. MODIFI assumes that each k-mer in a metagenome is mostly unmodified and calculates background signal levels for that k-mer from PacBio HiFi reads, eliminating the need for matched control experiments. MODIFI ECE-host linkages were validated using >1,000 isolate and mock microbiome datasets. Illustrating the approach, we identified 315 strain-resolved, non-redundant ECE-host linkages in environmental and human metagenomes. In infant gut microbiomes, a chromosomal inversion in Enterococcus faecalis alters host and associated plasmid methylation motifs simultaneously. Overall, MODIFI solves a major bottleneck in DNA modification analysis and provides a foundational tool for understanding microbial epigenomics.
]]></description>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Guitor, A. K.</dc:creator>
<dc:creator>Valentin-Alvarado, L. E.</dc:creator>
<dc:creator>Garner, R.</dc:creator>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Yan, M.</dc:creator>
<dc:creator>Shi, L.-D.</dc:creator>
<dc:creator>Schoelmerich, M. C.</dc:creator>
<dc:creator>Steininger, H. M.</dc:creator>
<dc:creator>Portik, D. M.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Wilkinson, J. E.</dc:creator>
<dc:creator>Lynch, S.</dc:creator>
<dc:creator>Morowitz, M. J.</dc:creator>
<dc:creator>Hess, M.</dc:creator>
<dc:creator>Diamond, S.</dc:creator>
<dc:creator>Banfield, J. F.</dc:creator>
<dc:creator>Sachdeva, R.</dc:creator>
<dc:date>2026-03-28</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714056</dc:identifier>
<dc:title><![CDATA[Metagenomic strain-resolved DNA modification patterns link extrachromosomal genetic elements to host strains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.26.713760v1?rss=1">
<title>
<![CDATA[
Spike-in probe-enhanced single-cell RNA-seq reveals post-infusion transcriptomic remodeling of "prime-and-kill" synNotch-CAR-T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.26.713760v1?rss=1</link>
<description><![CDATA[
We previously developed synthetic Notch (synNotch)-chimeric antigen receptor (CAR)-T cells to improve the safety and efficacy of CAR-T therapy for glioblastoma. In this system, an anti-EphA2/IL13R2-dual-CAR is expressed only upon recognition of tumor- or brain-specific "priming" antigens, EGFRvIII (termed E-SYNC cells) or brevican (B-SYNC), respectively, with E-SYNC currently under phase I clinical evaluation (NCT06186401). However, tracking and profiling these engineered cells in vivo remain challenging, limiting our understanding of their activity and therapeutic potential. To address this gap, we developed a single-cell RNA-sequencing (scRNA-seq) workflow with custom spike-in probes for synNotch-CAR transcripts, enabling simultaneous detection of engineered cells and transcriptomic profiling. In vitro, integration of multiple probes using machine-learning-assisted classifiers detected 78.2% of E-SYNC cells and 60.0% of B-SYNC cells with 98.0% specificity. In a xenograft model, synNotch-positive cells were detected across the spleen, lung, and brain, with the highest frequency and most robust priming and activation observed in the brain. Single-cell transcriptomic analyses revealed tissue-specific differentiation programs, including cytotoxicity, proliferation, metabolic activity, and acquisition of tissue-resident memory phenotypes, shaped by both environmental cues and synNotch-mediated antigen recognition. In summary, this spike-in probe-enhanced scRNA-seq workflow enables robust detection and high-resolution characterization of synNotch-CAR-T cell dynamics and provides a broadly applicable platform for monitoring engineered immune cells in diverse clinical contexts.

One Sentence SummaryOur spike-in probe-enhanced single-cell RNA-sequencing method enables analysis of tissue-dependent activation and transcriptional states of synNotch-CAR-T cells, providing a robust and scalable platform for in vivo tracking and transcriptomic profiling of engineered cell therapies.
]]></description>
<dc:creator>Nejo, T.</dc:creator>
<dc:creator>Watchmaker, P. B.</dc:creator>
<dc:creator>Simic, M. S.</dc:creator>
<dc:creator>Yamamichi, A.</dc:creator>
<dc:creator>Lakshmanachetty, S.</dc:creator>
<dc:creator>Zhao, A.</dc:creator>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>Gallus, M.</dc:creator>
<dc:creator>Benway, H. L.</dc:creator>
<dc:creator>Zhu, R.</dc:creator>
<dc:creator>Almeida, R.</dc:creator>
<dc:creator>Lim, W. A.</dc:creator>
<dc:creator>Okada, H.</dc:creator>
<dc:date>2026-03-29</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.713760</dc:identifier>
<dc:title><![CDATA[Spike-in probe-enhanced single-cell RNA-seq reveals post-infusion transcriptomic remodeling of "prime-and-kill" synNotch-CAR-T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.26.714483v1?rss=1">
<title>
<![CDATA[
StrataChip: a microphysiological system capturing dynamic keratinocyte fate and mechanical transitions during human epidermal morphogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.26.714483v1?rss=1</link>
<description><![CDATA[
Epidermal development and homeostasis require precise coordination between keratinocyte differentiation and mechanics. Still, the mechanisms integrating these processes remain poorly understood in part due to limitations of existing experimental systems. Here, we introduce StrataChip, a tractable microphysiological system that enables dynamic, multimodal interrogation of human epidermal morphogenesis. The platform integrates a media perfused dermal tissue with human epidermal keratinocytes within a microfluidic device and supports rapid epidermal stratification following establishment of an air-liquid interface. High-resolution confocal imaging and single-cell RNA-sequencing demonstrate that the StrataChip recapitulates key architectural and molecular features of human epidermis, including distinct basal, spinous, and granular layers defined by canonical differentiation markers and adhesion molecule organization. Single-cell profiling reveals transcriptionally distinct basal and spinous subpopulations, including transitional states associated with suprabasal commitment. Live 3D imaging in situ captures keratinocyte morphodynamics including basal cell delamination and asymmetric division, linking dynamic cellular behaviors to defined differentiation fates and stratification. Altogether, StrataChip provides a robust platform for a dynamic and mechanistic interrogation of how gene regulation and cell mechanics are coupled during epidermal morphogenesis.
]]></description>
<dc:creator>Amakor, J. K.</dc:creator>
<dc:creator>Rajan, A. A. N.</dc:creator>
<dc:creator>Kamaraj, M.</dc:creator>
<dc:creator>Jacobs, K. A.</dc:creator>
<dc:creator>Hutchins, E. J.</dc:creator>
<dc:creator>Wittmann, T.</dc:creator>
<dc:creator>Kutys, M. L.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.714483</dc:identifier>
<dc:title><![CDATA[StrataChip: a microphysiological system capturing dynamic keratinocyte fate and mechanical transitions during human epidermal morphogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.26.714555v1?rss=1">
<title>
<![CDATA[
Prenatal diet buffers infant epigenetic changes linked to pollution and transient wheeze 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.26.714555v1?rss=1</link>
<description><![CDATA[
Prenatal air pollution exposure is associated with childhood asthma, particularly among biological males. The mechanisms remain unclear, but may involve lasting epigenetic changes, such DNA methylation (DNAm), that occur during gestation in response to oxidative stress and inflammation. Higher maternal intake of "protective" micronutrients, like antioxidants, could buffer pollution-induced oxidative stress and inflammation to mitigate potentially adverse DNAm differences contributing to asthma. Using data from 515 CANDLE participants, we examined associations between prenatal NO2, PM2.5, and PM10 and cord blood DNAm, evaluated DNAm mediation of pollution associations with childhood wheeze phenotypes (transient, persistent, and late-onset), and assessed buffering of DNAm by maternal polyunsaturated fatty acid, vitamin C, or folate intake, and overall diet quality measured by the Alternative Healthy Eating Index-Pregnancy (AHEI-P). We identified 19, seven, and five regional DNAm differences associated NO2, PM2.5, and PM10. Mediation analyses suggested a role for HLA-DPA1/DPB1 DNAm in NO2 and PM2.5 associations with transient wheeze. To assess buffering, we fit pollutant-by-diet interaction models, defining buffering as an interaction opposite in sign to the main pollutant effect. One or more micronutrients or AHEI-P attenuated pollutant effects at 16 of 19 NO2-associated DMRs, including HLA-DPA1/DPB1, and all PM2.5- and PM10-associated DMRs. However, attenuation of HLA-DPA1/DPB1 DNAm did not significantly reduce the indirect effect of NO2 on transient wheeze. In sex-stratified analyses, biological males exhibited lower PM2.5-associated DNAm in SERPINB9, a gene linked to lung function. These findings suggest prenatal air pollution alters DNAm, which may contribute to transient wheeze, with some differences partially buffered by maternal diet.

Significance StatementPrenatal air pollution exposure contributes to child wheeze and asthma, potentially through the oxidative stress response and subsequent changes to infant DNA methylomes. Here, we used data from the CANDLE cohort to identify cord blood DNAm differences associated with NO2, PM2.5, or PM10. We examined if any alterations mediated the relationship between prenatal air pollution exposures and transient, persistent, or late-onset wheeze at age 4 to 6 years. Some of these DNAm differences appeared to be at least partially buffered by maternal micronutrients and/or overall diet quality.
]]></description>
<dc:creator>Lee, S. A.</dc:creator>
<dc:creator>Konwar, C.</dc:creator>
<dc:creator>Balshaw, R.</dc:creator>
<dc:creator>MacIsaac, J. L.</dc:creator>
<dc:creator>Ramadori, K.</dc:creator>
<dc:creator>Lin, D. T. S.</dc:creator>
<dc:creator>Urtatiz, O.</dc:creator>
<dc:creator>LeWinn, K. Z.</dc:creator>
<dc:creator>Karr, C. J.</dc:creator>
<dc:creator>Smith, A. K.</dc:creator>
<dc:creator>Kobor, M. S.</dc:creator>
<dc:creator>Carroll, K. N.</dc:creator>
<dc:creator>Bush, N. R.</dc:creator>
<dc:creator>Jones, M. J.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.714555</dc:identifier>
<dc:title><![CDATA[Prenatal diet buffers infant epigenetic changes linked to pollution and transient wheeze]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.30.715090v1?rss=1">
<title>
<![CDATA[
Multi-cohort Analysis Reveals Microbiome Signatures Associated with Drug Response in New-Onset Rheumatoid Arthritis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.30.715090v1?rss=1</link>
<description><![CDATA[
The human gut microbiome influences treatment outcomes, but whether microbiome signatures of drug response generalize across cohorts remains unclear. Here, we perform a multi-cohort analysis (3 cohorts, N=100 patients) to determine whether cross-cohort microbial signatures are associated with methotrexate (MTX) response in new-onset rheumatoid arthritis (RA) patients. Pre-treatment gut microbiome community structure and function differed by future MTX response status, with MTX-nonresponders (MTX-NR) showing Bacteroides caccae depletion and Ruminococcus bromii enrichment. Multiple microbial functions were depleted in MTX-NR, including nucleotide metabolism, one-carbon pool by folate, and histidine metabolism. Notably, candidate MTX-degrading genes were enriched in MTX-NR. Microbiome functional profiles outperformed clinical metrics in predicting future MTX response. These results show that consistent microbiome signatures are associated with MTX response across different RA cohorts and pave the way for microbiome-based precision medicine in newly diagnosed RA patients.
]]></description>
<dc:creator>Bodkhe, R. L.</dc:creator>
<dc:creator>Blank, R. B.</dc:creator>
<dc:creator>Trepka, K. R.</dc:creator>
<dc:creator>Orellana, D. A.</dc:creator>
<dc:creator>Scher, J. U.</dc:creator>
<dc:creator>Turnbaugh, P. J.</dc:creator>
<dc:creator>Nayak, R. R.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.30.715090</dc:identifier>
<dc:title><![CDATA[Multi-cohort Analysis Reveals Microbiome Signatures Associated with Drug Response in New-Onset Rheumatoid Arthritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.714805v1?rss=1">
<title>
<![CDATA[
Sleep and emotional memory: translating neural response at encoding to memory accuracy in men and women with and without PTSD 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714805v1?rss=1</link>
<description><![CDATA[
BackgroundGrowing evidence suggests that sleep plays an important role in PTSD outcomes, potentially due to its influence on emotional memory consolidation, though these mechanisms remain unknown. This study sought to test the hypotheses that sleep neurophysiology, PTSD status, and sex moderates the degree to which the late positive potential (LPP) mediates memory accuracy for affective visual stimuli.

MethodsN = 39 participants (18 female) viewed 75 negative and 75 neutral IAPS images while EEG was recorded. After viewing the images, participants took a two-hour long nap which was followed by a memory assessment. Memory accuracy was measured using d = Z(hit rate) - Z(false alarm rate), where hit rate refers to the proportion of images seen during the memory assessment that are correctly identified as being previously seen, false alarm rate refers to the proportion of images seen during the memory assessment that are incorrectly identified as being previously seen, and Z() is the inverse cumulative distribution function of the standard normal distribution function.

ResultsThe early (300 - 1000 ms) and late (1000 - 1500 ms) LPP mediated enhanced discrimination accuracy for emotional compared to neural stimuli (d) (ps < 0.001). The association between the late LPP and d was moderated by sleep such that the association was stronger when participants spent proportionately more time in N3 and REM (p = 0.02). The differences in reactivity between emotional and neutral images for both the early and late LPP were attenuated in PTSD+ individuals vs. controls (ps < 0.001). Despite mediation results showing greater d for emotional compared to neutral stimuli, women showed overall worse memory accuracy for negative compared to neutral stimuli (p < 0.001) whereas men showed no difference (p = 0.64).

ConclusionsN3 and REM sleep play a critical role for memory of stimuli that produce large and sustained neural responses. PTSD is marked by a diminished ability to distinguish between negative and neutral information. More research is critical to understand sex effects on emotional memory.
]]></description>
<dc:creator>Santistevan, A. C.</dc:creator>
<dc:creator>Natraj, N.</dc:creator>
<dc:creator>Yack, L. M.</dc:creator>
<dc:creator>Felmingham, K. L.</dc:creator>
<dc:creator>Woodward, S. H.</dc:creator>
<dc:creator>Mathalon, D. H.</dc:creator>
<dc:creator>Neylan, T. C.</dc:creator>
<dc:creator>Richards, A.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714805</dc:identifier>
<dc:title><![CDATA[Sleep and emotional memory: translating neural response at encoding to memory accuracy in men and women with and without PTSD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.714827v1?rss=1">
<title>
<![CDATA[
Cell state-specific metabolic networks govern ferroptosis versus apoptosis in small cell lung cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714827v1?rss=1</link>
<description><![CDATA[
Cellular heterogeneity and plasticity are hallmarks of cancer that contribute to tumor growth and therapy resistance. Here we investigated metabolic heterogeneity in small cell lung cancer (SCLC), an aggressive neuroendocrine (NE) cancer type. Through integrated transcriptomic and metabolomic analyses, we identified a universal dependency on exogenous cysteine/cystine (Cys) across all NE/non-NE SCLC cell states. Notably, NE and non-NE cells with low levels of the ASCL1 transcription factor die from ferroptosis upon Cys depletion. In contrast, ASCL1-high cells die from apoptosis but are ferroptosis resistant. This resistance to ferroptosis is driven by the direct upregulation of the gene coding for the GCH1 enzyme by ASCL1, which results in higher levels of the BH4/BH2 antioxidants. Accordingly, combining cysteine depletion with BH4/BH2 synthesis inhibition effectively reduces tumor growth in patient-derived xenografts. This work elucidates distinct metabolic states in SCLC and suggests new approaches to induce cell death in this lethal form of cancer.
]]></description>
<dc:creator>Kim, J. W.</dc:creator>
<dc:creator>Bebber, C. M.</dc:creator>
<dc:creator>Dai, Y.</dc:creator>
<dc:creator>Bopp, S.</dc:creator>
<dc:creator>Edenhofer, A.</dc:creator>
<dc:creator>Li, A. M.</dc:creator>
<dc:creator>Rosner, T.</dc:creator>
<dc:creator>Berning, L.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Leak, L. B.</dc:creator>
<dc:creator>Stroh, J.</dc:creator>
<dc:creator>Shrestha, B.</dc:creator>
<dc:creator>Abdallah, A.</dc:creator>
<dc:creator>Prymidis, D.</dc:creator>
<dc:creator>Olivos, H.</dc:creator>
<dc:creator>Baron, M.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Shue, Y. T.</dc:creator>
<dc:creator>Nishiga, Y.</dc:creator>
<dc:creator>Drainas, A.</dc:creator>
<dc:creator>Chaikovsky, A.</dc:creator>
<dc:creator>Szylo, K.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Kang, Y. P.</dc:creator>
<dc:creator>Manoj, P.</dc:creator>
<dc:creator>Quintanal Villalonga, A.</dc:creator>
<dc:creator>Rudin, C. M.</dc:creator>
<dc:creator>DeNicola, G. M.</dc:creator>
<dc:creator>Dixon, S. J.</dc:creator>
<dc:creator>Frezza, C.</dc:creator>
<dc:creator>Ye, J.</dc:creator>
<dc:creator>von Karstedt, S.</dc:creator>
<dc:creator>Sage, J.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714827</dc:identifier>
<dc:title><![CDATA[Cell state-specific metabolic networks govern ferroptosis versus apoptosis in small cell lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.28.715039v1?rss=1">
<title>
<![CDATA[
Neural correlates of novel word-form learning in developmental language disorder 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.28.715039v1?rss=1</link>
<description><![CDATA[
Children with developmental language disorder (DLD) have persistent language learning difficulties and often perform poorly on pseudoword repetition, a task that probes phonological, memory, and speech-motor processes that support vocabulary acquisition. Research on the neural basis of pseudoword repetition in DLD is limited. We used whole-brain functional MRI (fMRI) to examine pseudoword repetition and repetition-based learning in 46 children with DLD (ages 10-15 years) and 71 age-matched children with typical language development. During scanning, children heard and repeated pseudowords paired with visual referents, allowing us to track learning-related changes in neural activity across repetitions. Repeated pseudoword production yielded comparable behavioural learning across groups, with faster productions by later repetitions. Post-scan, form-referent recognition was comparable across groups, whereas pseudoword repetition accuracy was lower in DLD. Pseudoword repetition engaged a distributed neural network, including inferior frontal cortex bilaterally (greater on the left), premotor and sensorimotor cortex, and posterior temporal and occipital regions. Group differences emerged primarily in regions where activity was task negative (i.e., below baseline or deactivated): lateral occipito-parietal cortex (posterior angular gyrus), medial parieto-occipital cortex (retrosplenial), and right posterior cingulate cortex. Learning-related decreases in activity were similar across groups, but region-of-interest analyses showed reduced leftward lateralisation of activity in inferior frontal gyrus in DLD. These findings suggest weaker disengagement of the default mode network during a linguistically demanding task in DLD. Although repetition-based pseudoword learning recruited similar neural mechanisms in both groups, these mechanisms may operate less efficiently in DLD, alongside reduced hemispheric specialisation in inferior frontal cortex.

HighlightsO_LISimilar repetition-related neural attenuation across groups during pseudoword learning.
C_LIO_LIReduced default-mode network suppression during pseudoword repetition in DLD.
C_LIO_LIReduced left-hemisphere specialisation of inferior frontal cortex in DLD.
C_LIO_LIRepetition-based learning in DLD supported by less efficient neural networks.
C_LI
]]></description>
<dc:creator>Bahar, N.</dc:creator>
<dc:creator>Cler, G. J.</dc:creator>
<dc:creator>Asaridou, S. S.</dc:creator>
<dc:creator>Smith, H. J.</dc:creator>
<dc:creator>Willis, H. E.</dc:creator>
<dc:creator>Healy, M. P.</dc:creator>
<dc:creator>Chughtai, S.</dc:creator>
<dc:creator>Haile, M.</dc:creator>
<dc:creator>Krishnan, S.</dc:creator>
<dc:creator>Watkins, K. E.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.28.715039</dc:identifier>
<dc:title><![CDATA[Neural correlates of novel word-form learning in developmental language disorder]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.30.710191v1?rss=1">
<title>
<![CDATA[
Resolution of recursive data corruption to transform T-cell epitope discovery 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.30.710191v1?rss=1</link>
<description><![CDATA[
Accurate prediction of MHC class I-presented peptides is essential for any vaccine or T-cell therapy design, yet reported gains on in silico benchmarks have not translated into clinical successes. Here we show that this discrepancy may come from a common methodological error: immunopeptidomics datasets are fundamentally contaminated by existing prediction models through prediction-based deconvolution and filtering, resulting in an iterative confirmation bias. An audit of the IEDB, the biggest database in the field, reveals that as of January 2025, 55.8% of assessable data are labeled by computational models rather than verified experimentally. This inflates in silico benchmarks while degrading real-world applicability on new data, effectively making it impossible to objectively test model performance, which can lead to choosing suboptimal solutions and decreasing the chance of any therapys clinical success. In silico simulation shows that iterative data corruption maintains high AUROC while top-of-list retrieval collapses. We reframe epitope discovery as a protein-centric learning-to-rank task and introduce deepMHCflare, a model evaluated exclusively on clean data. deepMHCflare achieves 0.80 Precision@4 on mono-allelic benchmarks versus 0.55-0.65 for gold-standard prediction models. A preclinical cancer vaccine study validated that 2 of the 4 deepMHCflare-nominated peptides were immunogenic, with a third independently confirmed in the literature.
]]></description>
<dc:creator>Preibisch, G.</dc:creator>
<dc:creator>Tyrolski, M.</dc:creator>
<dc:creator>Kucharski, P.</dc:creator>
<dc:creator>Gizinski, S.</dc:creator>
<dc:creator>Grzegorczyk, P.</dc:creator>
<dc:creator>Moon, S.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Zaro, B.</dc:creator>
<dc:creator>Gambin, A.</dc:creator>
<dc:date>2026-04-01</dc:date>
<dc:identifier>doi:10.64898/2026.03.30.710191</dc:identifier>
<dc:title><![CDATA[Resolution of recursive data corruption to transform T-cell epitope discovery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.30.714220v1?rss=1">
<title>
<![CDATA[
An integrated single cell and spatial omics atlas of human prenatal development 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.30.714220v1?rss=1</link>
<description><![CDATA[
Single cell genomics has enabled analysis of human prenatal development at unprecedented resolution. However, most studies have relied on dissociated tissues during restricted windows of development, limiting insights into how spatially distributed networks of cells, and multicellular niches emerge and adapt to distinct organ microenvironments in situ. Moreover, existing human developmental atlases have not yet been harmonised, and we thus lack a comprehensive catalogue of known cell types in the developing human body.

Here, we introduce the Human Developmental Cell Atlas (HDCA), a unified structural, cellular and molecular resource for prenatal human development. The HDCA integrates published and unpublished single cell/nucleus RNAseq atlases across prenatal organs, and includes a newly generated, spatially resolved, multimodal cell atlas of intact human embryos. Spanning 4-22 post conceptional weeks, capturing embryonic and early to mid fetal stages, the HDCA contains [~]4.6 million cells/nuclei which resolve into [~]450 cell types, explorable with a bespoke web portal.

For a global overview of the human embryos multicellular communities, we applied unsupervised deep learning to our intact human embryo spatial data, charting 114 tissue niches that are structural and signalling hubs for the cellular interactions of the embryo. Guided by these niches, we profiled cellular networks over space and time, not examinable using single-organ atlases. In so doing, we revealed tissue-specific fibroblast patterning from previously undescribed mesenchyme progenitors, early diversification of organ-specific blood capillaries and lymphatic vasculature, emergence of neural crest cell fates, the formation of placode-and neural crest-derived peripheral sensory neurons, and how tissue niches guide peripheral neuron maturation and axonal migration. The HDCA thus serves as a comprehensive step towards a comprehensive understanding of human prenatal development, and a template towards unravelling the biology of congenital disorders.
]]></description>
<dc:creator>Webb, S.</dc:creator>
<dc:creator>Rose, A.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Steele, L.</dc:creator>
<dc:creator>Kuri, M. A.</dc:creator>
<dc:creator>Stephenson, E.</dc:creator>
<dc:creator>Inecik, K.</dc:creator>
<dc:creator>Jafree, D.</dc:creator>
<dc:creator>Foster, A. R.</dc:creator>
<dc:creator>Basurto-Lozada, D.</dc:creator>
<dc:creator>Chipampe, N.-J.</dc:creator>
<dc:creator>Pournara, A. V.</dc:creator>
<dc:creator>Jacques, M.-A.</dc:creator>
<dc:creator>To, K.</dc:creator>
<dc:creator>Admane, C.</dc:creator>
<dc:creator>Kritikaki, E.</dc:creator>
<dc:creator>Chroscik, M. A.</dc:creator>
<dc:creator>Horsfall, D.</dc:creator>
<dc:creator>Foreman, J.</dc:creator>
<dc:creator>Rademaker, K.</dc:creator>
<dc:creator>Karjalainen, J.</dc:creator>
<dc:creator>Laddach, A.</dc:creator>
<dc:creator>Madad, S.</dc:creator>
<dc:creator>Lawrence, J. E. G.</dc:creator>
<dc:creator>Kleshchevnikov, V.</dc:creator>
<dc:creator>Lisgo, S.</dc:creator>
<dc:creator>Lee, J. T. H.</dc:creator>
<dc:creator>Blevinal, J.</dc:creator>
<dc:creator>Alqahtani, A.</dc:creator>
<dc:creator>Makarchuk, S.</dc:creator>
<dc:creator>Jackson, J.</dc:creator>
<dc:creator>Ucuncu, E.</dc:creator>
<dc:creator>Silva, T. P.</dc:creator>
<dc:creator>Lorenzi, V.</dc:creator>
<dc:creator>Torabi, F.</dc:creator>
<dc:creator>Botting, R. A.</dc:creator>
<dc:creator>Roberts, K.</dc:creator>
<dc:creator>Olabi, B.</dc:creator>
<dc:creator>Chakala, K. P.</dc:creator>
<dc:creator>Dony, L.</dc:creator>
<dc:creator>Dall'Aglio, G.</dc:creator>
<dc:creator>Cujba, A.-M.</dc:creator>
<dc:creator>Whitfield, H. J.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2026-04-01</dc:date>
<dc:identifier>doi:10.64898/2026.03.30.714220</dc:identifier>
<dc:title><![CDATA[An integrated single cell and spatial omics atlas of human prenatal development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.30.715205v1?rss=1">
<title>
<![CDATA[
Population genomics reveals multi-scale mechanisms sustaining schistosomiasis re-emergence in a near-elimination setting 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.30.715205v1?rss=1</link>
<description><![CDATA[
In China, sustained snail control, environmental management, and mass drug administration with praziquantel reduced schistosomiasis to near-elimination levels, yet re-emergence in Sichuan Province during the early 2000s exposed vulnerabilities in late-stage control. We conducted a novel investigation of Schistosoma japonicum re-emergence in Sichuan in order to identify the processes underlying re-emergence and to showcase how genomic data can be used in such investigations. We sequenced whole genomes from 270 miracidia collected from 53 human hosts across 17 villages in 2007 - the year after re-emergence was documented.

Population genomic analyses revealed a broadly cohesive regional parasite population with weak geographic structuring. Genome-wide diversity was substantial, and demographic reconstructions revealed no recent decline in effective population size, demonstrating that parasite populations were not demographically fragmented at the onset of re-emergence, hinting at maintenance in reservoir populations. At finer spatial scales, several villages exhibited low diversity and elevated inbreeding, consistent with localized transmission maintained by small founding populations. Estimates of pairwise genetic relatedness revealed dense within-village sibling clusters alongside second- and third-degree relationships spanning villages, and rare first- and second-degree cross-village links. These findings are consistent with highly focal transmission with episodic parasite dispersal across villages, leading to a regional transmission network. Inferred minimum worm burden across hosts varied from 1 to 11 adult worm pairs, indicating heterogeneity in within-host parasite diversity, although uneven sampling limited inference. Together, these findings indicate that schistosomiasis re-emergence in this near-elimination setting was likely facilitated by a diverse parasite population maintained in reservoir populations, and that transmission, while predominantly local was occurring across a network of connected villages. This work illustrates how population genomics can reveal mechanisms driving re-emergence in late-stage elimination in complex, multi-host transmission systems.

Author SummarySchistosomiasis is a parasitic disease that affects millions of people worldwide. Understanding the factors that contribute to persistence and transmission, even in the face of control programs, is central to reducing and ultimately eliminating the disease. In China, decades of intensive control efforts have dramatically reduced infections, yet the disease continued to persist in some regions. In this study, we analyzed the genomes of 270 parasite larvae collected from infected humans across 17 villages in Sichuan Province, China, shortly after schistosomiasis re-emerged in the region. By examining genetic relationships among parasites, we reconstructed patterns of transmission across villages and within individual infections. Our findings suggest that, despite major reductions in human infection prevalence, parasite populations sizes do not show signatures of recent population decline, indicating that non-human hosts may serve as key reservoirs promoting persistence. We also find that local transmission pathways, including clonal infections infecting multiple humans derived from a relatively small number of snail hosts, maintain parasite transmission pathways. Collectively, these findings suggest non-human hosts, intact local transmission pathways, and regional connectivity together enabled re-emergence, highlighting important challenges for elimination programs
]]></description>
<dc:creator>Guss, H.</dc:creator>
<dc:creator>Francioli, Y.</dc:creator>
<dc:creator>Grover, E.</dc:creator>
<dc:creator>Hill, A.</dc:creator>
<dc:creator>Zou, W.</dc:creator>
<dc:creator>Wade, K.</dc:creator>
<dc:creator>Pike, H.</dc:creator>
<dc:creator>Gopalan, S. S.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Bo, Z.</dc:creator>
<dc:creator>Pollock, D.</dc:creator>
<dc:creator>Carlton, E.</dc:creator>
<dc:creator>Castoe, T. A.</dc:creator>
<dc:date>2026-04-01</dc:date>
<dc:identifier>doi:10.64898/2026.03.30.715205</dc:identifier>
<dc:title><![CDATA[Population genomics reveals multi-scale mechanisms sustaining schistosomiasis re-emergence in a near-elimination setting]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.30.715396v1?rss=1">
<title>
<![CDATA[
Temporal AI model predicts drivers of cell state trajectories across human aging 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.30.715396v1?rss=1</link>
<description><![CDATA[
Foundational AI models have recently shown promise for predicting the impact of perturbations on cell states. However, current models typically consider only one cell state at a time, limiting their ability to learn how cellular responses unfold over time, particularly across long trajectories such as diseases of aging. Here, we develop a temporal AI model, MaxToki, trained on nearly 1 trillion gene tokens including cell state trajectories across the human lifespan to generate cell states across long timelapses of human aging. MaxToki generalized to unseen trajectories through in-context learning and predicted novel age-modulating targets that were experimentally verified to influence age-related gene programs and functional decline in vivo. MaxToki represents a promising strategy for temporal modeling to accelerate the discovery of interventions for programming therapeutic cellular trajectories.
]]></description>
<dc:creator>Gomez Ortega, J.</dc:creator>
<dc:creator>Nadadur, R. D.</dc:creator>
<dc:creator>Kunitomi, A.</dc:creator>
<dc:creator>Kothen-Hill, S.</dc:creator>
<dc:creator>Wagner, J. U. G.</dc:creator>
<dc:creator>Kurtoglu, S. D.</dc:creator>
<dc:creator>Kim, B.</dc:creator>
<dc:creator>Reid, M. M.</dc:creator>
<dc:creator>Lu, T.</dc:creator>
<dc:creator>Washizu, K.</dc:creator>
<dc:creator>Zanders, L.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Ancheta, S.</dc:creator>
<dc:creator>Lichtarge, S.</dc:creator>
<dc:creator>Johnson, W. A.</dc:creator>
<dc:creator>Thompson, C.</dc:creator>
<dc:creator>Phan, D. M.</dc:creator>
<dc:creator>Combes, A. J.</dc:creator>
<dc:creator>Yang, A. C.</dc:creator>
<dc:creator>Tadimeti, N.</dc:creator>
<dc:creator>Dimmeler, S.</dc:creator>
<dc:creator>Yamanaka, S.</dc:creator>
<dc:creator>Alexanian, M.</dc:creator>
<dc:creator>Theodoris, C. V.</dc:creator>
<dc:date>2026-04-01</dc:date>
<dc:identifier>doi:10.64898/2026.03.30.715396</dc:identifier>
<dc:title><![CDATA[Temporal AI model predicts drivers of cell state trajectories across human aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.31.715474v1?rss=1">
<title>
<![CDATA[
State-Dependent Organization of Microscale Functional Circuitry in Visual Cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.31.715474v1?rss=1</link>
<description><![CDATA[
Brain state modulates sensory processing across visual cortex, yet how it relates to the organization of functional circuitry at the level of individual neurons and cell types remains largely unknown. To address this, we constructed one of the largest mi-croscale directed functional circuit maps in mouse visual cortex from calcium imaging of more than 57,000 neurons across four visual areas and five cortical layers. Using a time-aware causal inference framework, we found that intra-areal connections dom-inate across arousal states, consistent with experimental findings on the local bias of cortical anatomy. Among intra-areal connections, anterolateral area (AL) had the highest density, and among inter-areal connections, the AL{leftrightarrow}rostrolateral area (RL) axis formed the strongest pathway. Laminar circuit organization was dominated by layer 6 recurrence within-layer, while the most prominent between-layer pathway was layer 5-to-layer 6 in low arousal and layer 4-to-layer 5 in high arousal. Spatial extent was selectively greater for excitatory-to-inhibitory connections in high arousal, but not for excitatory-to-excitatory connections. Across 6,597 electron-microscopy recon-structions of neuron pairs, synapse count predicted functional connection strength in both arousal states, but structure-function coupling was weaker in high arousal. In stimulus-driven response prediction, neuron pairs with stronger functional connections exhibited more similar predictive performance in both states, with performance vary-ing by layer and cell type. Overall, our findings map, at single-neuron resolution, the multi-scale organization of directed functional circuitry in mouse visual cortex across brain states.
]]></description>
<dc:creator>Biswas, R.</dc:creator>
<dc:creator>Wickrama Senevirathne, H.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Abbasi-Asl, R.</dc:creator>
<dc:date>2026-04-01</dc:date>
<dc:identifier>doi:10.64898/2026.03.31.715474</dc:identifier>
<dc:title><![CDATA[State-Dependent Organization of Microscale Functional Circuitry in Visual Cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.31.714901v1?rss=1">
<title>
<![CDATA[
Aerobic exercise prevents the loss of endogenous pain modulation in male and female rats with traumatic brain injury. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.31.714901v1?rss=1</link>
<description><![CDATA[
Traumatic Brain Injury (TBI) patients may suffer from a number of long-term complications after injury such as impaired motor skills, cognitive decline, and sensory abnormalities including chronic pain. Disruption of endogenous pain modulatory pathways likely contributes to development of chronic pain in a wide range of conditions including TBI. Aerobic exercise has been shown to impact pain syndromes. Here we investigate the effect of exercise on pain outcome measures after TBI using a lateral fluid percussion (LFP) model and voluntary running wheels in male and female rats. We tested mechanical nociceptive reactivity with von Frey fibers and descending control of nociception (DCN) using hindpaw sensitization with PGE2 followed by a capsaicin-test stimulus to the forepaw. Pharmacological studies employed the administration of noradrenergic (NA) and serotoninergic receptor blockers. Neuropathological studies quantified neuroinflammatory changes and axonal damage. We found that exercise decreased the duration of the acute phase of pain from [~]5 weeks to 2-3 weeks in female and male TBI rats respectively, gains that could be reversed using the 1-adrenoceptor (1AR) antagonist, prazosin. Exercise also prevented the loss of DCN for at least 180 days post-injury in both male and female TBI rats. The intact DCN response in male and female TBI rats provided by exercise could be blocked using prazosin. Surprisingly, exercise-mediated restoration of the DCN response in male TBI rats was not blocked by the 5-HT7 receptor antagonist, SB-267790, the receptor system through which serotonin reuptake inhibitors restore DCN after TBI in male rats. Therefore, the transition from a noradrenergic to a serotonergic inhibitory pain pathway that we typically see in male TBI rats, was blocked by exercise. Assessment of neuropathology, acutely after TBI, reveals that both the astrocyte and microglial response to injury is significantly greater in male TBI compared to female TBI, regardless of exercise. The effect of exercise on the extent of neuroinflammation after injury was minimal in TBI rats of both sexes. In contrast, exercise significantly decreased the amount of axonal loss in the corpus callosum in both male and female TBI rats compared to sedentary TBI rats. However, the extent of axonal loss after TBI in both exercise and sedentary male rats was greater than in female exercise and sedentary groups respectively. These results demonstrate that exercise is a promising treatment for chronic pain after TBI in both male and females. It also highlights that dysfunction of the endogenous pain modulatory pathways observed in male rats after TBI can be prevented by exercise, possibly by reducing axonal loss.
]]></description>
<dc:creator>Irvine, K.-A.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Clark, D. J.</dc:creator>
<dc:date>2026-04-02</dc:date>
<dc:identifier>doi:10.64898/2026.03.31.714901</dc:identifier>
<dc:title><![CDATA[Aerobic exercise prevents the loss of endogenous pain modulation in male and female rats with traumatic brain injury.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.31.710877v1?rss=1">
<title>
<![CDATA[
Epilepsy-associated potassium channel KCNT1 is required for multiciliated cell development in Xenopus 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.31.710877v1?rss=1</link>
<description><![CDATA[
Pathogenic variants in the gene KCNT1, which encodes a sodium-activated potassium channel, cause a severe neurodevelopmental disorder with intractable epilepsy. In addition to seizures, affected individuals commonly present with severe respiratory issues and structural heart defects not commonly observed in other genetic pediatric epilepsies, suggesting additional developmental functions for KCNT1 in organs beyond the brain. Here, we characterized the spectrum of clinical diagnoses present in a cohort of 46 individuals with pathogenic variants in KCNT1, ranging from 0 to 19 years of age, by medical record review. We documented the prevalence of diagnoses across organ systems, including dependence on assisted breathing, congenital structural heart defects, urinary dysfunction, and spine deformities, among others. Next, we explored the embryonic expression and function of KCNT1 in diploid frogs (Xenopus tropicalis) and observed expression in developing ciliated tissues such as the brain, heart, kidney, and epidermis. Embryonic perturbation of KCNT1 disrupted developmental signaling pathways and caused ciliogenesis defects in the mucociliary epidermis, a common model for the human airway. Loss of KCNT1 disrupted development of multiciliated cells, reminiscent of recent work on the ion channel Piezo1. Consistently, pharmacological inhibition of Piezo signaling enhanced the ciliogenesis phenotype observed following KCNT1 inhibition, while activation of Piezo1 activity partially rescued ciliogenesis in the context of KCNT1 inhibition. Together, this work establishes that KCNT1 has embryonic functions in Xenopus beyond regulating neuronal activity, specifically in multiciliated cell development, and identifies an interaction with pharmacologically-tractable Piezo channels that may be productive for therapeutic efforts.
]]></description>
<dc:creator>Chemel, A. K.</dc:creator>
<dc:creator>McCluskey, K. E.</dc:creator>
<dc:creator>Tran, M. N.</dc:creator>
<dc:creator>Ehrlich, A. T.</dc:creator>
<dc:creator>Willsey, H. R.</dc:creator>
<dc:date>2026-04-03</dc:date>
<dc:identifier>doi:10.64898/2026.03.31.710877</dc:identifier>
<dc:title><![CDATA[Epilepsy-associated potassium channel KCNT1 is required for multiciliated cell development in Xenopus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.31.715694v1?rss=1">
<title>
<![CDATA[
Molecular dynamics of Brodmann Area 22 in development and autism 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.31.715694v1?rss=1</link>
<description><![CDATA[
Challenges in verbal communication are a prominent feature of autism. However, gene regulatory programs in speech-related cortical regions remain poorly characterized. In parallel, it remains unclear whether the heterogeneous genetic factors underlying autism converge on shared neurobiological mechanisms. To address these gaps, we generated paired transcriptomic and epigenomic data from post-mortem human brain tissue across 100 donors. Here, we show that transcriptional differences in the speech-related Brodmann Area 22 in individuals with neurodevelopmental conditions, including autism, are strongest among those with a known genetic diagnosis. A similar but attenuated signature is observed in those without a genetic diagnosis. These transcriptional differences are most pronounced in neurons, with glutamatergic L4/5 intratelencephalic neurons affected across multiple modalities. Finally, multimodal analysis implicates altered RFX3-dependent networks as a central hub in autism, particularly among L4/5 intratelencephalic neurons in non-verbal individuals. Together, our study identifies regulatory architecture linking chromatin state, transcriptional output, and variation in verbal ability in autism.
]]></description>
<dc:creator>Suresh, V.</dc:creator>
<dc:creator>Wigdor, E. M.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Leonard, R.</dc:creator>
<dc:creator>Asfouri, J.</dc:creator>
<dc:creator>Griffiths, M.</dc:creator>
<dc:creator>Evans, C.</dc:creator>
<dc:creator>Yuan, G.</dc:creator>
<dc:creator>Rohani, N.</dc:creator>
<dc:creator>Weiss, J.</dc:creator>
<dc:creator>Dema, C.</dc:creator>
<dc:creator>Mukhthar, T.</dc:creator>
<dc:creator>Lassen, F.</dc:creator>
<dc:creator>Schafer, N.</dc:creator>
<dc:creator>Dong, S.</dc:creator>
<dc:creator>Palmer, D. S.</dc:creator>
<dc:creator>Chang, E. F.</dc:creator>
<dc:creator>Sanders, S. J.</dc:creator>
<dc:creator>Nowakowski, T. J.</dc:creator>
<dc:date>2026-04-03</dc:date>
<dc:identifier>doi:10.64898/2026.03.31.715694</dc:identifier>
<dc:title><![CDATA[Molecular dynamics of Brodmann Area 22 in development and autism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.04.02.714281v1?rss=1">
<title>
<![CDATA[
Dietary tryptophan mitigates lung ischemia-reperfusion injury via microbiota-derived indole-3-propionate and aryl hydrocarbon receptor signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.02.714281v1?rss=1</link>
<description><![CDATA[
BackgroundLung ischemia-reperfusion (IR) injury drives early morbidity after lung transplantation and cardiothoracic surgery, yet targeted preventive therapies are lacking. The gut-lung axis and microbiota-derived tryptophan metabolites, including indole-3-propionate (IPA), may regulate pulmonary immunity and inflammation. We investigated whether a tryptophan-rich (Trp-Rich) diet attenuates sterile lung IR injury by increasing microbiota-derived indole metabolites and reprogramming alveolar macrophage (AM) inflammatory responses.

MethodsC57BL/6 mice received isocaloric tryptophan-standard (Trp-Std; 0.18%) or Trp-Rich (0.60%) diets for 14 days, then underwent unilateral left lung IR (60 min ischemia followed by 60 min reperfusion). Oxygen saturation, lung cytokines, and aryl hydrocarbon receptor (AhR) signaling readouts (Cyp1a1/Cyp1b1) were evaluated. Gut microbiota was profiled by 16S rRNA sequencing, and targeted metabolomics quantified tryptophan metabolites in feces, portal vein (PV) plasma, and lung tissue. To further assess inflammatory priming in vivo, mice were additionally challenged with intratracheal lipopolysaccharide (LPS). Mechanistic studies compared IPA with related indole metabolites in MH-S cells and primary human AMs, including ex vivo nutritional IR, LPS stimulation, and AhR stimulation and blockade using synthetic agonists and antagonists.

ResultsTrp-Rich feeding improved post-IR oxygenation, reduced lung IL-1{beta}, and increased pulmonary Cyp1a1/Cyp1b1 gene expression. Trp-Rich diet remodeled the gut microbiota, including enrichment of Bifidobacterium and Lactobacillus, and increased IPA levels across feces, PV plasma, and lung tissue, with lower kynurenine/IPA ratios across matrices. In the LPS intratracheal challenge, Trp-Rich feeding reduced IL-6 levels in lung tissue and systemic plasma. Primary murine AMs isolated from Trp-Rich mice also showed reduced IL-1{beta} and IL-6 release in an ex vivo nutritional IR model. Among tested indole metabolites, IPA showed the strongest dose-dependent suppression of LPS-induced cytokines and chemokines in MH-S cells and primary human AMs, remained active in the ex vivo nutritional IR model, and its anti-inflammatory effect was abrogated by AhR blockade and enhanced by co-treatment with other indole metabolites.

ConclusionsA Trp-Rich diet attenuated sterile lung IR injury, coinciding with gut microbiota remodeling, increased systemic and pulmonary IPA, reduced inflammatory priming, and reprogrammed AM responses. These data support diet- or microbiome-directed strategies targeting IPA-AhR signaling to mitigate perioperative lung IR injury.

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O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=190 SRC="FIGDIR/small/714281v1_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@112753org.highwire.dtl.DTLVardef@2cdd0eorg.highwire.dtl.DTLVardef@164e413org.highwire.dtl.DTLVardef@b9a34a_HPS_FORMAT_FIGEXP  M_FIG A tryptophan-rich diet remodels the gut microbiota and indole metabolite profiles, including IPA, enhances alveolar macrophage AhR signaling, and attenuates sterile lung ischemia-reperfusion injury.

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]]></description>
<dc:creator>Chaki, T.</dc:creator>
<dc:creator>Maruyama, D.</dc:creator>
<dc:creator>Doan, T. N.</dc:creator>
<dc:creator>Tian, X.</dc:creator>
<dc:creator>Prakash, A.</dc:creator>
<dc:date>2026-04-06</dc:date>
<dc:identifier>doi:10.64898/2026.04.02.714281</dc:identifier>
<dc:title><![CDATA[Dietary tryptophan mitigates lung ischemia-reperfusion injury via microbiota-derived indole-3-propionate and aryl hydrocarbon receptor signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.04.02.716136v1?rss=1">
<title>
<![CDATA[
Gain-of-function mutation in SKAP2 leads to type 1 diabetes and broader autoimmunity through hyperactive integrin signaling in myeloid cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.02.716136v1?rss=1</link>
<description><![CDATA[
Many genetic variants associated with increased type 1 diabetes (T1D) risk are located within the SKAP2 gene; however, the mechanisms by which these variants confer disease risk remain unclear. SKAP2 encodes an adapter protein that functions within the integrin signaling pathway and is found at the highest levels in myeloid leukocytes. We recently identified a de novo gain-of-function SKAP2 mutation in an individual with T1D, leading to hyperactive integrin signaling in myeloid cells. To dissect the mechanisms by which this mutation may lead to T1D, we generated a knock-in mouse line containing the orthologous p.G153R substitution in mouse SKAP2 on the diabetes-prone nonobese diabetic (NOD) genetic background. Both female and male SKAP2G153R/G153R mice developed accelerated T1D. The SKAP2G153R/G153R mice also exhibited a unique spectrum of autoantibodies, leading to immune-complex nephritis. Accelerated infiltration of pancreatic islets by myeloid cells, B lymphocytes, and activated T cells was observed in SKAP2G153R/G153R mice. Single-cell RNA sequencing demonstrated a type 1 IFN{gamma}-driven inflammatory program within the pancreatic islets of SKAP2G153R/G153R mice. Dendritic cells from SKAP2G153R/G153R mice demonstrated increased antigen-presenting capacity, characterized by enhanced adhesion to T cells during immune synapse formation. Macrophages and neutrophils from SKAP2G153R/G153R mice also showed increased integrin signaling responses, with neutrophils expressing high levels of activated {beta}2 integrins on the cell surface. When backcrossed onto the C57BL/6J genetic background, the SKAP2G153R/G153R mice developed spontaneous autoantibody formation and exhibited accelerated autoimmunity, including nephritis, in the pristane-induced model of autoimmune disease. These findings demonstrate that dysregulation of leukocyte integrin signaling, through alterations in SKAP2, may increase the genetic risk for autoimmunity and T1D.
]]></description>
<dc:creator>Tamaki, C. M.</dc:creator>
<dc:creator>Chamberlain, C. E.</dc:creator>
<dc:creator>Abram, C. L.</dc:creator>
<dc:creator>Poojary, S.</dc:creator>
<dc:creator>Bridge, J.</dc:creator>
<dc:creator>Matsuda, J. L.</dc:creator>
<dc:creator>Tamaki, W.</dc:creator>
<dc:creator>Rutsch, N.</dc:creator>
<dc:creator>Spector, L.</dc:creator>
<dc:creator>Dixon, W.</dc:creator>
<dc:creator>Proekt, I.</dc:creator>
<dc:creator>Letourneau-Freiberg, L. R.</dc:creator>
<dc:creator>Philipson, L. H.</dc:creator>
<dc:creator>German, M. S.</dc:creator>
<dc:creator>Anderson, M. S.</dc:creator>
<dc:creator>Lowell, C. A.</dc:creator>
<dc:date>2026-04-06</dc:date>
<dc:identifier>doi:10.64898/2026.04.02.716136</dc:identifier>
<dc:title><![CDATA[Gain-of-function mutation in SKAP2 leads to type 1 diabetes and broader autoimmunity through hyperactive integrin signaling in myeloid cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.04.04.715981v1?rss=1">
<title>
<![CDATA[
FunctionaL Assigning Sequence Homing (FLASH) maps phenotype to sequence with deep and machine learning 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.04.715981v1?rss=1</link>
<description><![CDATA[
Genome-wide association studies (GWAS) map genetic variation to a reference genome and correlate variants to phenotypes. Yet, GWAS and similar procedures have limitations, including an inability to predict phenotype on variants never seen during the discovery phase and difficulty integrating structural variants. Deep and machine learning alternatives have not been successful at consistent prediction of resistance phenotypes (Hu et al. 2024). Here, we introduce FLASH: a new interpretable, statistically-based deep learning framework that operates directly on raw sequencing reads. In over 35,000 isolates of bacteria, fungi and viruses, FLASH achieves uniformly high accuracy on independent test data, including on variation never seen in training, meeting or exceeding bespoke state of the art methods. FLASH identifies canonical drug targets ab initio and new pan-species predictors of virulence, including those lacking annotation and those only partially aligned to NCBI reference databases. Further, FLASH can predict phenotypes beyond the possibility of GWAS, such as bacterial host range of phage, a task that to our knowledge is impossible today. FLASH is simple to run, highly efficient and constitutes a new approach for predicting gene function and phenotype across the tree of life. It is especially valuable when bioethical concerns and the vast genetic complexity of pathogenic microbes limit the feasibility of experimental validation.
]]></description>
<dc:creator>Cotter, D. J.</dc:creator>
<dc:creator>Harrison, M.-C.</dc:creator>
<dc:creator>Rustagi, A.</dc:creator>
<dc:creator>Wang, P. L.</dc:creator>
<dc:creator>Kokot, M.</dc:creator>
<dc:creator>Carey, A. F.</dc:creator>
<dc:creator>Deorowicz, S.</dc:creator>
<dc:creator>Salzman, J.</dc:creator>
<dc:date>2026-04-07</dc:date>
<dc:identifier>doi:10.64898/2026.04.04.715981</dc:identifier>
<dc:title><![CDATA[FunctionaL Assigning Sequence Homing (FLASH) maps phenotype to sequence with deep and machine learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.04.04.716514v1?rss=1">
<title>
<![CDATA[
Tau-induced mitochondrial reverse electron transport drives neurodegeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.04.716514v1?rss=1</link>
<description><![CDATA[
Hyperphosphorylation and aggregation of the microtubule-associated protein tau are recognized as pathological hallmarks of tauopathies; however, the biological activity of tau that drives its pathophysiological effects remains poorly understood1-6. Mitochondrial dysfunction is a common feature of tauopathies7,8. Despite this, the mechanistic link between tau abnormalities and mitochondrial dysfunction, as well as its relationship to taus physiological function, remains unclear. Here, we demonstrate that tau regulates mitochondrial reverse electron transport (RET), which produces excess ROS, reduces the NAD+/NADH ratio, and is activated by aging or stress. In flies, mice, and human induced pluripotent stem cells (hiPSC)-derived neurons, tau depletion eliminates stress-induced RET and confers significant stress resistance. Mechanistically, tau enters mitochondria and directly interacts with the mitochondrial complex I (C-I) subunit NDUFS3, enhancing RET activation in a phosphorylation-dependent manner that correlates with tau pathogenicity. Elevated RET further drives tau hyperphosphorylation, establishing a self-perpetuating pathological loop. Blocking tau entry into mitochondria or disrupting tau/NDUFS3 interaction reduces tau-induced RET. Genetic or pharmacological inhibition of RET protects against tau-induced neurodegeneration across species. RET regulation represents a previously unrecognized normal function of tau that becomes pathological in disease, providing a therapeutic target for conditions characterized by tau abnormalities and mitochondrial dysfunction.
]]></description>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Rimal, S.</dc:creator>
<dc:creator>Bhurtel, S.</dc:creator>
<dc:creator>Yeung, L.</dc:creator>
<dc:creator>Lu, B.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Cobos, I.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Guo, S.</dc:creator>
<dc:creator>Lu, B.</dc:creator>
<dc:date>2026-04-07</dc:date>
<dc:identifier>doi:10.64898/2026.04.04.716514</dc:identifier>
<dc:title><![CDATA[Tau-induced mitochondrial reverse electron transport drives neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.04.05.716562v1?rss=1">
<title>
<![CDATA[
Gut microbiome-dependent IL-1 signaling is a mediator of ACVR1R206H-driven heterotopic ossification 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.05.716562v1?rss=1</link>
<description><![CDATA[
Inflammatory diseases cause significant morbidity and mortality, but their pathobiology is often difficult to dissect due to complex genetic-environmental interactions. Genetic forms of heterotopic ossification, such as fibrodysplasia ossificans progressiva (FOP), reduce genetic variability, allowing careful dissection of non-genetic drivers of inflammation. While >95% of FOP patients harbor the ACVR1R206H mutation, patients exhibit significant variability in disease progression, suggesting a role of environmental drivers. Here, we identify the gut microbiome as a regulator of inflammation-driven HO in FOP. Metagenomic profiling of cohabitating FOP/unaffected sibling pairs revealed a pathogenic gut microbiome profile in FOP patients (Bray-Curtis, p < 0.05). In Pdgfr-Cre/Acvr1R206H (FOP) mice, gut microbiome ablation by antibiotics reduced spontaneous HO formation (47.4% reduction, p < 0.05) and reduced plasma IL-1 pathway activity. IL-1{beta} blockade in FOP mice suppressed trauma-induced HO formation. These findings identify a gut microbiome-IL-1-HO axis with modifiable targets for developing treatments for HO and related inflammatory conditions.

One Sentence SummaryAntibiotic disruption of the gut microbiome reduces HO in FOP mice via an IL-1 mediated pathway.
]]></description>
<dc:creator>Herzog, H. M.</dc:creator>
<dc:creator>Fang, C.</dc:creator>
<dc:creator>Lam, L.</dc:creator>
<dc:creator>Jin, K.</dc:creator>
<dc:creator>Zamarioli, A.</dc:creator>
<dc:creator>Dinh, E.</dc:creator>
<dc:creator>Gupta, C. L.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Moody, T.</dc:creator>
<dc:creator>Pierce, J. L.</dc:creator>
<dc:creator>Hohl, M. S.</dc:creator>
<dc:creator>Takimoto, S. W.</dc:creator>
<dc:creator>Lyalina, S.</dc:creator>
<dc:creator>Wentworth, K. L.</dc:creator>
<dc:creator>Yu, K.</dc:creator>
<dc:creator>Lu, V. F.</dc:creator>
<dc:creator>Isadora Mamikunian, I.</dc:creator>
<dc:creator>Hunt, N. K.</dc:creator>
<dc:creator>Lynch, S.</dc:creator>
<dc:creator>Pollard, K. S.</dc:creator>
<dc:creator>Hernandez, C. J.</dc:creator>
<dc:creator>Perrien, D. S.</dc:creator>
<dc:creator>Hsiao, E. C.</dc:creator>
<dc:date>2026-04-08</dc:date>
<dc:identifier>doi:10.64898/2026.04.05.716562</dc:identifier>
<dc:title><![CDATA[Gut microbiome-dependent IL-1 signaling is a mediator of ACVR1R206H-driven heterotopic ossification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.04.06.716539v1?rss=1">
<title>
<![CDATA[
Successful dendritic cell vaccines require lasting in-situ TNF α secretion to license antitumor CD8 + T cell cytotoxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.06.716539v1?rss=1</link>
<description><![CDATA[
Dendritic cells (DCs) are central to activating cytotoxic CD8 T cells, yet DC-based vaccines have achieved limited success against established tumors. To address this gap, we analyzed the transcriptomic and functional changes CD8 T cells undergo following interactions with DC subsets in lymphoid organs and tumor sites. This approach allowed us to map their trajectory from naive to fully cytotoxic effector cells. We found that classical DCs in lymphoid organs provide essential antigen presentation but fail to elicit cytotoxicity. Instead, antigenexperienced CD8 T cells require additional inflammatory signals, primarily through TNF, delivered at tumor sites by infiltrating myeloid DCs. Effective cytotoxic responses therefore depend on the synchronization of these distinct, temporally separated signals. Notably, tumor antigen-pulsed DC vaccines rapidly lose TNF expression after infiltrating tumors, limiting their efficacy. These findings establish a sequential model of T cell activation and suggest strategies to enhance the potency of DC-based immunotherapies.
]]></description>
<dc:creator>Khateeb, A. R.</dc:creator>
<dc:creator>Magal, N. S.</dc:creator>
<dc:creator>Inbal, K.</dc:creator>
<dc:creator>Gleiberman, A.</dc:creator>
<dc:creator>Kaminitz, A.</dc:creator>
<dc:creator>Weiss, T.</dc:creator>
<dc:creator>Verbin, G.</dc:creator>
<dc:creator>Richter, A.</dc:creator>
<dc:creator>Zarfin, A.</dc:creator>
<dc:creator>Younis, L. F.</dc:creator>
<dc:creator>Gutwillig, A.</dc:creator>
<dc:creator>Frish, A.</dc:creator>
<dc:creator>Shifrut, E.</dc:creator>
<dc:creator>Reuveni, I. R.</dc:creator>
<dc:creator>Barzel, A.</dc:creator>
<dc:creator>Levi, C.</dc:creator>
<dc:creator>Rider, P.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:creator>Engleman, E. G.</dc:creator>
<dc:creator>Madi, A.</dc:creator>
<dc:creator>Carmi, Y.</dc:creator>
<dc:date>2026-04-08</dc:date>
<dc:identifier>doi:10.64898/2026.04.06.716539</dc:identifier>
<dc:title><![CDATA[Successful dendritic cell vaccines require lasting in-situ TNF α secretion to license antitumor CD8 + T cell cytotoxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
