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<title>bioRxiv Channel: Fred Hutchinson Cancer Center</title>
<link>https://biorxiv.org</link>
<description>
This feed contains articles for bioRxiv Channel "Fred Hutchinson Cancer Center"
</description>

<items>
<rdf:Seq>
</rdf:Seq>
</items>
<prism:eIssn/>
<prism:publicationName>bioRxiv</prism:publicationName>
<prism:issn/>

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</channel>
<image rdf:about="">
<title>bioRxiv</title>
<url/>
<link>https://biorxiv.org</link>
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<item rdf:about="https://biorxiv.org/content/10.1101/106054v1?rss=1">
<title>
<![CDATA[
The Rosetta all-atom energy function for macromolecular modeling and design 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/106054v1?rss=1</link>
<description><![CDATA[
Over the past decade, the Rosetta biomolecular modeling suite has informed diverse biological questions and engineering challenges ranging from interpretation of low-resolution structural data to design of nanomaterials, protein therapeutics, and vaccines. Central to Rosettas success is the energy function: amodel parameterized from small molecule and X-ray crystal structure data used to approximate the energy associated with each biomolecule conformation. This paper describes the mathematical models and physical concepts that underlie the latest Rosetta energy function, beta_nov15. Applying these concepts,we explain how to use Rosetta energies to identify and analyze the features of biomolecular models.Finally, we discuss the latest advances in the energy function that extend capabilities from soluble proteins to also include membrane proteins, peptides containing non-canonical amino acids, carbohydrates, nucleic acids, and other macromolecules.
]]></description>
<dc:creator>Alford, R. F.</dc:creator>
<dc:creator>Leaver-Fay, A.</dc:creator>
<dc:creator>Jeliazkov, J. R.</dc:creator>
<dc:creator>O'Meara, M. J.</dc:creator>
<dc:creator>DiMaio, F. P.</dc:creator>
<dc:creator>Park, H.</dc:creator>
<dc:creator>Shapovalov, M. V.</dc:creator>
<dc:creator>Renfrew, P. D.</dc:creator>
<dc:creator>Mulligan, V. K.</dc:creator>
<dc:creator>Kappel, K.</dc:creator>
<dc:creator>Labonte, J. W.</dc:creator>
<dc:creator>Pacella, M. S.</dc:creator>
<dc:creator>Bonneau, R.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Dunbrack, R. L.</dc:creator>
<dc:creator>Das, R.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:creator>Kuhlman, B.</dc:creator>
<dc:creator>Kortemme, T.</dc:creator>
<dc:creator>Gray, J. J.</dc:creator>
<dc:date>2017-02-07</dc:date>
<dc:identifier>doi:10.1101/106054</dc:identifier>
<dc:title><![CDATA[The Rosetta all-atom energy function for macromolecular modeling and design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.25.449763v1?rss=1">
<title>
<![CDATA[
BayesDeBulk: A Flexible Bayesian Algorithm for the Deconvolution of Bulk Tumor Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.25.449763v1?rss=1</link>
<description><![CDATA[
To understand immune activation and evasion mechanisms in cancer, one crucial step is to characterize the composition of immune and stromal cells in the tumor microenvironment (TME). Deconvolution analysis based on bulk transcriptomic data has been used to estimate cell composition in TME. However, these algorithms are sub-optimal for proteomic data, which has hindered research in the rapidly growing field of proteogenomics. Moreover, with the increasing prevalence of multi-omics studies, there is an opportunity to enhance deconvolution analysis by utilizing paired proteomic and transcriptomic profiles of the same tissue samples. To bridge these gaps, we propose BayesDeBulk, a new method for estimating the immune/stromal cell composition based on bulk proteomic and gene expression data. BayesDeBulk utilizes the information of known cell-type-specific markers without requiring their absolute abundance levels as prior knowledge. We compared BayesDeBulk with existing tools on synthetic and real data examples, demonstrating its superior performance and versatility.

AvailabilitySoftware available at http://www.BayesDeBulk.com/

ContactFor any information, please contact francesca.petralia@mssm.edu
]]></description>
<dc:creator>Petralia, F.</dc:creator>
<dc:creator>Calinawan, A. P.</dc:creator>
<dc:creator>Feng, S.</dc:creator>
<dc:creator>Gosline, S. J.</dc:creator>
<dc:creator>Pugliese, P.</dc:creator>
<dc:creator>Ceccarelli, M.</dc:creator>
<dc:creator>Wang, P.</dc:creator>
<dc:date>2021-06-25</dc:date>
<dc:identifier>doi:10.1101/2021.06.25.449763</dc:identifier>
<dc:title><![CDATA[BayesDeBulk: A Flexible Bayesian Algorithm for the Deconvolution of Bulk Tumor Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.08.451691v1?rss=1">
<title>
<![CDATA[
Multifactorial chromatin regulatory landscapes at single cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.08.451691v1?rss=1</link>
<description><![CDATA[
Chromatin profiling at locus resolution uncovers gene regulatory features that define cell types and developmental trajectories, but it remains challenging to map and compare distinct chromatin-associated proteins within the same sample. Here we describe a scalable antibody barcoding approach for profiling multiple chromatin features simultaneously in the same individual cells, Multiple Target Identification by Tagmentation (MulTI-Tag). MulTI-Tag is optimized to retain high sensitivity and specificity of enrichment for multiple chromatin targets in the same assay. We use MulTI-Tag in a combinatorial barcoding approach to resolve distinct cell types and developmental trajectories using multiple chromatin features, and to distinguish unique, coordinated patterns of active and repressive element regulatory usage in the same individual cells that are associated with distinct differentiation outcomes. Multifactorial profiling allows us to detect novel associations between histone marks in single cells and holds promise for comprehensively characterizing cell-specific gene regulatory landscapes in development and disease.
]]></description>
<dc:creator>Meers, M. P.</dc:creator>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2021-07-09</dc:date>
<dc:identifier>doi:10.1101/2021.07.08.451691</dc:identifier>
<dc:title><![CDATA[Multifactorial chromatin regulatory landscapes at single cell resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.16.472014v1?rss=1">
<title>
<![CDATA[
Dynamic structural cell responses in the thymus to acute injury, regeneration, and age 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.16.472014v1?rss=1</link>
<description><![CDATA[
The thymus is essential for establishing adaptive immunity yet undergoes age-related atrophy leading to compromised immune responsiveness. The thymus is also extremely sensitive to acute insult and although capable of regeneration, this capacity declines with age. Focusing on non-hematopoietic stromal cells, and using single-cell and spatial transcriptomics, lineage-tracing, and advanced imaging, we discovered two atypical thymic epithelial cell (TEC) states that emerged with age. Age-associated (aa)TECs formed atypical high-density epithelial clusters that were devoid of thymocytes, an accretion of non-functional thymic tissue that worsened with age and exhibited features of partial epithelial-to-mesenchymal transition (EMT). In silico interaction analysis revealed that aaTEC emergence drew tonic signals from other TEC populations at baseline, acting as a sink for TEC growth factors. Following damage, aaTEC expanded substantially, further perturbing trophic pathways, and correlating with defective regeneration of the involuted thymus. These findings define a unique feature of thymic involution linked to immune aging.
]]></description>
<dc:creator>Jahn, L. L.</dc:creator>
<dc:creator>Kousa, A. I.</dc:creator>
<dc:creator>Sikkema, L.</dc:creator>
<dc:creator>Flores, A. E.</dc:creator>
<dc:creator>Argyropoulos, K. V.</dc:creator>
<dc:creator>Tsai, J.</dc:creator>
<dc:creator>Lazrak, A.</dc:creator>
<dc:creator>Nichols, K.</dc:creator>
<dc:creator>Lee, N.</dc:creator>
<dc:creator>Malard, F.</dc:creator>
<dc:creator>Andrlova, H.</dc:creator>
<dc:creator>Gomes, A. L.</dc:creator>
<dc:creator>Velardi, E.</dc:creator>
<dc:creator>Youssef, S.</dc:creator>
<dc:creator>da Silva, M. B.</dc:creator>
<dc:creator>Docampo, M.</dc:creator>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Mazoutis, L.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:creator>Pe'er, D.</dc:creator>
<dc:creator>Manley, N. R.</dc:creator>
<dc:creator>Dudakov, J. A.</dc:creator>
<dc:creator>van den Brink, M. R.</dc:creator>
<dc:date>2021-12-17</dc:date>
<dc:identifier>doi:10.1101/2021.12.16.472014</dc:identifier>
<dc:title><![CDATA[Dynamic structural cell responses in the thymus to acute injury, regeneration, and age]]></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/10.1101/2022.03.17.484768v1?rss=1">
<title>
<![CDATA[
KAT5 activity regulates G0-like states in human gliomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.17.484768v1?rss=1</link>
<description><![CDATA[
In solid tumors, G0-like states are likely critical for maintaining developmental hierarchies and cellular heterogeneity and promoting tumor growth/recurrence, yet little is known about tumor G0 states or regulation of their ingress/egress. To discover G0-like states and their regulators for glioblastoma (GBM), we analyzed G0 populations in an orthotopic model of GBM using single cell RNA-seq and performed a genome-wide CRISPR-Cas9 screen in patient-derived GBM stem-like cells (GSCs) for genes that trap cells in G0 when inhibited. We identify the protein acetyltransferase KAT5 as a key regulator of transcriptional, epigenetic, and proliferative heterogeneity impacting transitions into G0-like states. KAT5 activity suppresses the emergence of non-dividing subpopulations with oligodendrocyte progenitor and radial glial cell characteristics both in vitro and in a human GSC brain tumor model. In primary gliomas, KAT5 activity is dynamic with KAT5low tumor cells displaying quiescent properties, while KAT5 activity overall increases from low to high grade tumors and is associated with worse patient outcomes.
]]></description>
<dc:creator>Mihalas, A.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>O'Connor, S.</dc:creator>
<dc:creator>Feldman, H.</dc:creator>
<dc:creator>Bassett, J.</dc:creator>
<dc:creator>Mitchell, K.</dc:creator>
<dc:creator>Jin, K.</dc:creator>
<dc:creator>Hoellerbauer, P.</dc:creator>
<dc:creator>Ling, M.</dc:creator>
<dc:creator>Jenkins, W.</dc:creator>
<dc:creator>Kufeld, M.</dc:creator>
<dc:creator>Corrin, P.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Aronow, B. J.</dc:creator>
<dc:creator>Plaisier, C. J.</dc:creator>
<dc:creator>Patel, A.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:date>2022-03-19</dc:date>
<dc:identifier>doi:10.1101/2022.03.17.484768</dc:identifier>
<dc:title><![CDATA[KAT5 activity regulates G0-like states in human gliomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.03.486888v1?rss=1">
<title>
<![CDATA[
A paracrine circuit of IL-1b/IL-1R1 between myeloid and tumor cells drives glioblastoma progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.03.486888v1?rss=1</link>
<description><![CDATA[
Monocytes and monocyte-derived macrophages (MDM) from blood circulation infiltrate and promote glioblastoma growth. Here we discover that glioma cells induce the expression of potent pro-inflammatory cytokine IL-1{beta} in MDM, which engages IL-1R1 in glioma cells, activates NF-{kappa}B pathway, and subsequently leads to the induction of monocyte chemoattractant proteins (MCPs). Thus, a feedforward paracrine circuit of IL-1{beta}/IL-1R1 between the tumors and MDM creates an interdependence driving glioblastoma progression. Locally antagonizing IL-1{beta}/IL-1R1 leads to reduced MDM infiltration, diminished tumor growth, reduced exhausted CD8+ T cells, and thereby extends the survival of tumor-bearing mice. In contrast to IL-1{beta}, IL-1a exhibits anti-tumor effects. Genetic deletion of Il1a is associated with decreased recruitment of lymphoid cells and loss of interferon (IFN) signaling in various immune populations and subsets of malignant cells. IL-1{beta} antagonism of IL-1{beta} should be considered as an effective anti-glioblastoma therapy.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=196 HEIGHT=200 SRC="FIGDIR/small/486888v1_ufig1.gif" ALT="Figure 1">
View larger version (66K):
org.highwire.dtl.DTLVardef@1c99e57org.highwire.dtl.DTLVardef@1ba9d24org.highwire.dtl.DTLVardef@a2c418org.highwire.dtl.DTLVardef@1f8091c_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Giotti, B.</dc:creator>
<dc:creator>Kaluzova, M.</dc:creator>
<dc:creator>Herting, C.</dc:creator>
<dc:creator>Pinero, G.</dc:creator>
<dc:creator>vallcorba, m. p.</dc:creator>
<dc:creator>Cristea, S.</dc:creator>
<dc:creator>Ross, J. L.</dc:creator>
<dc:creator>Ackley, J.</dc:creator>
<dc:creator>Maximov, V.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:creator>Marquez-Ropero, M.</dc:creator>
<dc:creator>Angione, A.</dc:creator>
<dc:creator>Nichols, N.</dc:creator>
<dc:creator>Tsankkova, N.</dc:creator>
<dc:creator>Michor, F.</dc:creator>
<dc:creator>Shayakhmetov, D. M.</dc:creator>
<dc:creator>Gutmann, D. H.</dc:creator>
<dc:creator>Tsankov, A. M.</dc:creator>
<dc:creator>Hambardzumyan, D.</dc:creator>
<dc:date>2022-04-05</dc:date>
<dc:identifier>doi:10.1101/2022.04.03.486888</dc:identifier>
<dc:title><![CDATA[A paracrine circuit of IL-1b/IL-1R1 between myeloid and tumor cells drives glioblastoma progression]]></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/10.1101/2022.04.14.488397v1?rss=1">
<title>
<![CDATA[
Cell wound repair requires the coordinated action of linear and branched actin nucleation factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.14.488397v1?rss=1</link>
<description><![CDATA[
Cells are subjected to a barrage of daily insults that often lead to its cortex being ripped open and requiring immediate repair. An important component of the cell s repair response is the formation of an actomyosin ring at the wound periphery to mediate its closure. Inhibition of linear actin nucleation factors and myosin result in a disrupted contractile apparatus and delayed wound closure. Here we show that branched actin nucleators function as a scaffold to assemble and maintain this contractile actomyosin cable. Removing branched actin leads to the formation of smaller circular actin-myosin structures at the cell cortex and slow wound closure. Removing linear and branched actin results in failed wound closure. Surprisingly, removal of branched actin and myosin results in the formation of parallel linear actin filaments that undergo a chiral swirling movement to close the wound. These results provide insight into actin organization in contractile actomyosin rings and uncover a new mechanism of wound closure.

SummaryHui et al. find that branched actin is required during cell wound repair to serve as a scaffold to anchor the contractile actomyosin cable at the wound periphery. Inhibition of branched actin and myosin results in parallel linear filaments that swirl to close the wound, uncovering a new mechanism for cell wound repair.
]]></description>
<dc:creator>Hui, J.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Dubrulle, J.</dc:creator>
<dc:creator>Parkhurst, S. M.</dc:creator>
<dc:date>2022-04-14</dc:date>
<dc:identifier>doi:10.1101/2022.04.14.488397</dc:identifier>
<dc:title><![CDATA[Cell wound repair requires the coordinated action of linear and branched actin nucleation factors]]></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/10.1101/2022.04.29.489989v1?rss=1">
<title>
<![CDATA[
Robust Normalization and Integration of Single-cell Protein Expression across CITE-seq Datasets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.29.489989v1?rss=1</link>
<description><![CDATA[
CITE-seq enables paired measurement of surface protein and mRNA expression in single cells using antibodies conjugated to oligonucleotide tags. Due to the high copy number of surface protein molecules, sequencing antibody-derived tags (ADTs) allows for robust protein detection, improving cell-type identification. However, variability in antibody staining leads to batch effects in the ADT expression, obscuring biological variation, reducing interpretability, and obstructing cross-study analyses. Here, we present ADTnorm (https://github.com/yezhengSTAT/ADTnorm), a normalization and integration method designed explicitly for ADT abundance. Benchmarking against 14 existing scaling and normalization methods, we show that ADTnorm accurately aligns populations with negative- and positive-expression of surface protein markers across 13 public datasets, effectively removing technical variation across batches and improving cell-type separation. ADTnorm enables efficient integration of public CITE-seq datasets, each with unique experimental designs, paving the way for atlas-level analyses. Beyond normalization, ADTnorm includes built-in utilities to aid in automated threshold-gating as well as assessment of antibody staining quality for titration optimization and antibody panel selection. Applying ADTnorm to a published COVID-19 CITE-seq dataset allowed for identifying previously undetected disease-associated markers, illustrating a broad utility in biological applications.
]]></description>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Jun, S.-H.</dc:creator>
<dc:creator>Tian, Y.</dc:creator>
<dc:creator>Florian, M.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:date>2022-05-01</dc:date>
<dc:identifier>doi:10.1101/2022.04.29.489989</dc:identifier>
<dc:title><![CDATA[Robust Normalization and Integration of Single-cell Protein Expression across CITE-seq Datasets]]></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/10.1101/2022.05.02.490302v1?rss=1">
<title>
<![CDATA[
Profiling RNA at chromatin targets in situ by antibody-targeted tagmentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.02.490302v1?rss=1</link>
<description><![CDATA[
Whereas techniques to map chromatin-bound proteins are well-developed, mapping chromatin-associated RNAs remains a challenge. Here we describe Reverse Transcribe & Tagment (RT&Tag), in which RNAs associated with a chromatin epitope are targeted by an antibody followed by a protein A-Tn5 transposome. Localized reverse transcription generates RNA/cDNA hybrids that are subsequently tag-mented for sequencing by Tn5. We demonstrate the utility of RT&Tag in Drosophila cells for capturing the noncoding RNA roX2 with the dosage compensation complex and maturing transcripts associated with silencing histone modifications. We also show that RT&Tag can detect N6-methyladenosine (m6A)-modified mRNAs, and show that genes producing methylated transcripts are characterized by extensive promoter pausing of RNA polymerase II. The high efficiency of in situ antibody tethering and tagmentation makes RT&Tag especially suitable for rapid low-cost profiling of chromatin-associated RNAs from small samples.
]]></description>
<dc:creator>Khyzha, N.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:date>2022-05-03</dc:date>
<dc:identifier>doi:10.1101/2022.05.02.490302</dc:identifier>
<dc:title><![CDATA[Profiling RNA at chromatin targets in situ by antibody-targeted tagmentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.02.490337v1?rss=1">
<title>
<![CDATA[
Expression of YAP1-MAML2 and constitutively active YAP1 drive the formation of meningioma-like tumors in mice that resemble NF2-mutant meningiomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.02.490337v1?rss=1</link>
<description><![CDATA[
YAP1 is a transcriptional co-activator regulated by the Hippo Signaling Pathway, including NF2. Meningiomas are the most common primary brain tumors, a large percentage exhibit heterozygous loss of chromosome 22 (harboring the NF2 gene) and functional inactivation of the remaining NF2 copy, implicating oncogenic YAP activity in these tumors. Recently, fusions between YAP1 and MAML2 have been identified in a subset of pediatric NF2-wild type meningiomas. Here, we show that human YAP1-MAML2-positive meningiomas resemble NF2-mutant meningiomas by global and YAP-related gene expression signatures. We then show that expression of YAP1-MAML2 in mice induces tumors that resemble human YAP1 fusion-positive and NF2-mutant meningiomas by gene expression. We demonstrate that YAP1-MAML2 primarily functions by exerting TEAD-dependent YAP activity that is resistant to Hippo signaling. Treatment with YAP-TEAD inhibitors is sufficient to inhibit the viability of YAP1-MAML2-driven mouse tumors ex vivo. Finally, we show that expression of constitutively active YAP1 (S127/397A-YAP1) is sufficient to induce similar tumors suggesting that the YAP component of the gene fusion is the critical driver of these tumors. In summary, our results implicate YAP1-MAML2 as a causal oncogenic driver and highlight TEAD-dependent YAP activity as an oncogenic driver in YAP1-MAML2-fusion meningioma as well as NF2-mutant meningioma in general.
]]></description>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Arakaki, A.</dc:creator>
<dc:creator>Sievers, P.</dc:creator>
<dc:creator>Bonnin, D. A. A.</dc:creator>
<dc:creator>Paddison, P. J.</dc:creator>
<dc:creator>Sahm, F.</dc:creator>
<dc:creator>Cimino, P. J.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:date>2022-05-02</dc:date>
<dc:identifier>doi:10.1101/2022.05.02.490337</dc:identifier>
<dc:title><![CDATA[Expression of YAP1-MAML2 and constitutively active YAP1 drive the formation of meningioma-like tumors in mice that resemble NF2-mutant meningiomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.16.492156v1?rss=1">
<title>
<![CDATA[
Targeting advanced prostate cancer with STEAP1 chimeric antigen receptor T cell therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.16.492156v1?rss=1</link>
<description><![CDATA[
Six transmembrane epithelial antigen of the prostate 1 (STEAP1) is a compelling tumor-associated cell surface antigen for therapeutic targeting in solid tumors. We identified broad expression of STEAP1 (87% positive) in lethal metastatic prostate cancer, even more so than prostate-specific membrane antigen (PSMA, 60% positive) which is a clinically established diagnostic and therapeutic target. Second-generation chimeric antigen receptor (CAR) T cells were engineered for reactivity against STEAP1 and demonstrated substantial antitumor activity in metastatic human prostate cancer models in immunodeficient mice. Adoptive transfer of STEAP1 CAR T cells was associated with prolonged peripheral persistence and either disease eradication or substantial tumor growth inhibition with progressive disease demonstrating antigen loss. As STEAP1 CAR T cells were also highly active in antigen density conditions as low as [~]1,500 molecules/cell, we generated a human STEAP1 (hSTEAP1) knock-in (KI) mouse to evaluate the potential for on-target off-tumor toxicities. hSTEAP1-KI mice demonstrated a pattern of systemic hSTEAP1 expression akin to that observed in humans with the greatest expression found in the prostate gland. Mouse-in-mouse studies of STEAP1 CAR T cell therapy in immunocompetent hSTEAP1-KI mice engrafted with disseminated mouse prostate cancer showed preliminary safety without evidence of gross toxicity, cytokine storm, or architectural disruption and increased T cell infiltration at sites of systemic hSTEAP1 expression. Tumor responses and extension of survival were appreciated but antigen loss was identified in recurrent and progressive disease. In summary, we report the extent of STEAP1 expression in treatment-refractory metastatic prostate cancer, the generation of a STEAP1 CAR T cell therapy with promising potency and safety in preclinical studies of advanced prostate cancer, and antigen escape as a mechanism of resistance to effective STEAP1 CAR T cell therapy.
]]></description>
<dc:creator>Bhatia, V.</dc:creator>
<dc:creator>Kamat, N. V.</dc:creator>
<dc:creator>Pariva, T. E.</dc:creator>
<dc:creator>Wu, L.-T.</dc:creator>
<dc:creator>Tsao, A.</dc:creator>
<dc:creator>Sasaki, K.</dc:creator>
<dc:creator>Wiest, L. T.</dc:creator>
<dc:creator>Zhang, A.</dc:creator>
<dc:creator>Rudoy, D.</dc:creator>
<dc:creator>Gulati, R.</dc:creator>
<dc:creator>Patel, R. A.</dc:creator>
<dc:creator>Roudier, M. P.</dc:creator>
<dc:creator>True, L. D.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Priceman, S. J.</dc:creator>
<dc:creator>Ishihara, J.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:date>2022-05-17</dc:date>
<dc:identifier>doi:10.1101/2022.05.16.492156</dc:identifier>
<dc:title><![CDATA[Targeting advanced prostate cancer with STEAP1 chimeric antigen receptor T cell therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.17.492375v1?rss=1">
<title>
<![CDATA[
Liver humanized NSG-PiZ mice support the study of chronic hepatitis B virus infection and antiviral therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.17.492375v1?rss=1</link>
<description><![CDATA[
Hepatitis B virus (HBV) is a pathogen of major public health importance that is largely incurable once a chronic hepatitis B (CHB) infection is established. Only humans and great apes are fully permissive to HBV replication, and this species restriction has impacted HBV research by limiting the utility of small animal models of HBV. To combat the species restriction of HBV and enable more HBV studies in vivo, liver-humanized mouse models have been developed that harbor primary human hepatocytes (PHH) and are fully permissive to HBV infection and replication. Unfortunately, these models can be difficult to establish and are expensive commercially, which has limited their academic use. As an alternative mouse model to study HBV, we evaluated liver-humanized NSG-PiZ mice and showed that they are fully permissive to HBV and can develop CHB. Mice were infected with a precore mutant clinical isolate that has now been serially passaged through 3 generations of mice without loss of fitness. HBV selectively replicates in hCK18+ human hepatocytes within chimeric livers, and HBV+ mice secrete infectious virions and HBsAg into blood, while also harboring covalently closed circular DNA (cccDNA). HBV+ mice remain viremic for at least 169 days, which should enable the study of new curative therapies targeting CHB and respond to antiviral entecavir therapy. The extended duration of viremia is sufficient to enable the study of established and new therapeutic approaches targeting CHB. Furthermore, HBV+ PHH in NSG-PiZ mice can be transduced by the hepatotropic AAV3b and AAV.LK03 vector capsids, which should enable the study of curative gene therapies that target CHB. In summary, our data demonstrates that liver humanized NSG-PiZ mice can be used as a robust and cost-effective alternative to existing CHB models and may enable more academic research labs to study HBV disease pathogenesis and antiviral therapy in a setting that is fully permissive to ongoing replication.
]]></description>
<dc:creator>Colon-Thillet, R.</dc:creator>
<dc:creator>Stone, D.</dc:creator>
<dc:creator>Loprieno, M. A.</dc:creator>
<dc:creator>Klouser, L.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Santo, T. K.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Stensland, L.</dc:creator>
<dc:creator>Upham, S. L.</dc:creator>
<dc:creator>Pepper, G.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Aubert, M.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:date>2022-05-17</dc:date>
<dc:identifier>doi:10.1101/2022.05.17.492375</dc:identifier>
<dc:title><![CDATA[Liver humanized NSG-PiZ mice support the study of chronic hepatitis B virus infection and antiviral therapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.20.492866v1?rss=1">
<title>
<![CDATA[
Oncogenic KRAS alters splicing factor phosphorylation and alternative splicing in lung cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.20.492866v1?rss=1</link>
<description><![CDATA[
BackgroundAlternative RNA splicing is widely dysregulated in cancers including lung adenocarcinoma, where aberrant splicing events are frequently caused by somatic splice site mutations or somatic mutations of splicing factor genes. However, the majority of mis-splicing in cancers is unexplained by these known mechanisms. We hypothesize that the aberrant Ras signaling characteristic of lung cancers plays a role in promoting the alternative splicing observed in tumors.

MethodsWe recently performed transcriptome and proteome profiling of human lung epithelial cells ectopically expressing oncogenic KRAS and another cancer-associated Ras GTPase, RIT1. Unbiased analysis of phosphoproteome data identified altered splicing factor phosphorylation in KRAS-mutant cells, so we performed differential alternative splicing analysis using rMATS to identify significantly altered isoforms in lung epithelial. To determine whether these isoforms were uniquely regulated by KRAS, we performed a large-scale splicing screen in which we generated over 300 unique RNA sequencing profiles of isogenic A549 lung adenocarcinoma cells ectopically expressing 75 different wild-type or variant alleles across 28 genes implicated in lung cancer.

ResultsMass spectrometry data showed widespread downregulation of splicing factor phosphorylation in lung epithelial cells expressing mutant KRAS compared to cells expressing wild-type KRAS. We observed alternative splicing in the same cells, with 2196 and 2416 skipped exon events in KRASG12V and KRASQ61H cells, respectively, 997 of which were shared (p < 0.001 by hypergeometric test). In the high-throughput splicing screen, mutant KRAS induced the greatest number of differential alternative splicing events, second only to the RNA binding protein RBM45 and its mutant allele RBM45M126I. We identified ten high confidence cassette exon events across multiple KRAS variants and cell lines. These included differential splicing of the Myc Associated Zinc Finger (MAZ). As MAZ regulates expression of KRAS, this splice variant may be a mechanism for the cell to modulate wild-type KRAS levels in the presence of oncogenic KRAS.

ConclusionProteomic and transcriptomic profiling of lung epithelial cells uncovered splicing factor phosphorylation and mRNA splicing events regulated by oncogenic KRAS. These data suggest that in addition to widespread transcriptional changes, Ras signaling pathways in cancer promote post-transcriptional splicing changes that may contribute to oncogenic processes.
]]></description>
<dc:creator>Lo, A.</dc:creator>
<dc:creator>McSharry, M.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:date>2022-05-20</dc:date>
<dc:identifier>doi:10.1101/2022.05.20.492866</dc:identifier>
<dc:title><![CDATA[Oncogenic KRAS alters splicing factor phosphorylation and alternative splicing in lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.30.493973v1?rss=1">
<title>
<![CDATA[
Chemical degradation of BTK/TEC as a novel approach to inhibit platelet function and thrombosis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.30.493973v1?rss=1</link>
<description><![CDATA[
The tyrosine kinase BTK plays an important role in platelet function downstream of GPVI and CLEC2 receptors and has been proposed as a novel target to prevent thrombosis in patients that are at increased risk. However, current clinically approved BTK inhibitors have off target effects and are associated with an increased bleeding risk. In this study, we therefore explored whether BTK can be targeted for degradation in human platelets by using recently developed heterobifunctional molecules that employ the proteasomal system to break down BTK. Here we confirm that human platelets are highly susceptible to BTK degraders with the generic tyrosine kinase degrader TL12-186, and the BTK degraders DD-04-15 and DD-03-171 leading to breakdown of BTK and its closely related kinase TEC, an effect that was prevented by proteasomal inhibitors. Tandem Mass Tag proteomic analysis confirmed high selectivity with TL12-186 degrading BTK/TEC, FAK/PYK2 and FER, whereas DD-04-15 and DD-03-171 degraded BTK/TEC only. GPVI-mediated platelet integrin IIb{beta}3 activation, P-selectin expression, and phosphatidyl-serine exposure were largely impaired upon BTK/TEC degradation, with PAR-1-mediated responses left intact. This is the first study to demonstrate that chemical protein degraders can be successfully employed in anucleate human platelets to modulate their function.
]]></description>
<dc:creator>Trory, J. S.</dc:creator>
<dc:creator>Munkacsi, A.</dc:creator>
<dc:creator>Sledz, K.</dc:creator>
<dc:creator>Goudswaard, L. J.</dc:creator>
<dc:creator>Heesom, K. J.</dc:creator>
<dc:creator>Moore, S. F.</dc:creator>
<dc:creator>Nabet, B.</dc:creator>
<dc:creator>Aggarwal, V. K.</dc:creator>
<dc:creator>Hers, I.</dc:creator>
<dc:date>2022-05-30</dc:date>
<dc:identifier>doi:10.1101/2022.05.30.493973</dc:identifier>
<dc:title><![CDATA[Chemical degradation of BTK/TEC as a novel approach to inhibit platelet function and thrombosis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.06.494757v1?rss=1">
<title>
<![CDATA[
Gene amplification acts as a molecular foothold to facilitate cross-species adaptation and evasion of multiple antiviral pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.06.494757v1?rss=1</link>
<description><![CDATA[
Cross-species spillover events are responsible for many of the pandemics in human history including COVID-19; however, the evolutionary mechanisms that enable these events are poorly understood. We have previously modeled this process using a chimeric vaccinia virus expressing the rhesus cytomegalovirus-derived PKR antagonist RhTRS1 in place of its native PKR antagonists; E3L and K3L (VACV{Delta}E{Delta}K+RhTRS1). Using this virus, we demonstrated that gene amplification of rhtrs1 occurred early during experimental evolution and was sufficient to fully rescue virus replication in partially resistant African green monkey (AGM) fibroblasts. Notably, this rapid gene amplification also allowed limited virus replication in otherwise completely non-permissive human fibroblasts, suggesting that gene amplification may act as a "molecular foothold" to facilitate viral adaptation to multiple species. In this study, we demonstrate that there are multiple barriers to VACV{Delta}E{Delta}K+RhTRS1 replication in human cells, mediated by both PKR and RNase L. We experimentally evolved three AGM-adapted virus populations in human fibroblasts. Each population adapted to human cells bimodally, via an initial 10-fold increase in replication after only two passages followed by a second 10-fold increase in replication by passage nine. Using our Illumina-based pipeline, we found that some SNPs which had evolved during the prior AGM adaptation were rapidly lost, while 13 singlebase substitutions and short indels increased over time, including two SNPs unique to HFF adapted populations. Many of these changes were associated with components of the viral RNA polymerase, although no variant was shared between all three populations. Taken together, our results demonstrate that rhtrs1 amplification was sufficient to increase viral tropism after passage in an "intermediate species" and subsequently enabled the virus to adopt different, species-specific adaptive mechanisms to overcome distinct barriers to viral replication in AGM and human cells.
]]></description>
<dc:creator>Banerjee, S.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Geballe, A. P.</dc:creator>
<dc:creator>Rothenburg, S.</dc:creator>
<dc:creator>Kitzman, J. O.</dc:creator>
<dc:creator>Brennan, G.</dc:creator>
<dc:date>2022-06-08</dc:date>
<dc:identifier>doi:10.1101/2022.06.06.494757</dc:identifier>
<dc:title><![CDATA[Gene amplification acts as a molecular foothold to facilitate cross-species adaptation and evasion of multiple antiviral pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.13.496036v1?rss=1">
<title>
<![CDATA[
Helicobacter pylori chronic infection selects for effective colonizers of metaplastic glands 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.13.496036v1?rss=1</link>
<description><![CDATA[
Chronic gastric infection with Helicobacter pylori can lead to progressive tissue changes that culminate in cancer, but how H. pylori adapts to the changing tissue environment during disease development is not fully understood. In a transgenic mouse gastric metaplasia model, we found that strains from unrelated individuals differed in their ability to infect the stomach, to colonize metaplastic glands, and to induce proliferation and alter the expression of metaplasia-associated proteins. H. pylori isolates from different stages of disease from a single individual had differential ability to colonize healthy and metaplastic gastric glands. Exposure to the metaplastic environment selected for high gastric colonization by one of these strains. Complete genome sequencing revealed a unique alteration in the frequency of a variant allele of the putative adhesin sabB, arising from a recombination event with the related sialic acid binding adhesin (SabA) gene. Mutation of sabB strongly reduced adherence to both normal and metaplastic gastric tissue in multiple strain backgrounds and highly attenuated stomach colonization. Thus, the changing gastric environment during disease development promotes bacterial adhesin gene variation associated with enhanced gastric colonization.

ImportanceChronic infection with Helicobacter pylori is the primary risk factor for developing stomach cancer. As disease progresses H. pylori must adapt to a changing host tissue environment that includes induction of new cell fates in the cells that line the stomach. We tested representative H. pylori isolates collected from the same patient during early and later stages of disease in a mouse model where we can rapidly induce disease-associated tissue changes. Only the later-stage H. pylori strains could robustly colonize the diseased stomach environment. We also found that the ability to colonize the diseased stomach was associated with genetic variation in a putative cell surface adhesin gene called sabB. Additional experiments revealed that SabB promotes binding to stomach tissue and is critical for stomach colonization by the late-stage strains. Thus, H. pylori diversifies its genome during disease progression and these genomic changes highlight critical factors for bacterial persistence.
]]></description>
<dc:creator>O'Brien, V. P.</dc:creator>
<dc:creator>Jackson, L. K.</dc:creator>
<dc:creator>Frick, J.</dc:creator>
<dc:creator>Rodriguez Martinez, A. E.</dc:creator>
<dc:creator>Jones, D. S.</dc:creator>
<dc:creator>Johnston, C. D.</dc:creator>
<dc:creator>Salama, N. R.</dc:creator>
<dc:date>2022-06-15</dc:date>
<dc:identifier>doi:10.1101/2022.06.13.496036</dc:identifier>
<dc:title><![CDATA[Helicobacter pylori chronic infection selects for effective colonizers of metaplastic glands]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.21.496879v1?rss=1">
<title>
<![CDATA[
Nucleosome patterns in circulating tumor DNA reveal transcriptional regulation of advanced prostate cancer phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.21.496879v1?rss=1</link>
<description><![CDATA[
Advanced prostate cancers comprise distinct phenotypes, but tumor classification remains clinically challenging. Here, we harnessed circulating tumor DNA (ctDNA) to study tumor phenotypes by ascertaining nucleosome positioning patterns associated with transcription regulation. We sequenced plasma ctDNA whole genomes from patient-derived xenografts representing a spectrum of androgen receptor active (ARPC) and neuroendocrine (NEPC) prostate cancers. Nucleosome patterns associated with transcriptional activity were reflected in ctDNA at regions of genes, promoters, histone modifications, transcription factor binding, and accessible chromatin. We identified the activity of key phenotype-defining transcriptional regulators from ctDNA, including AR, ASCL1, HOXB13, HNF4G, and NR3C1. Using these features, we designed a prediction model which distinguished NEPC from ARPC in patient plasma samples across three clinical cohorts with 97-100% sensitivity and 85-100% specificity. While phenotype classification is typically assessed by immunohistochemistry or transcriptome profiling, we demonstrate that ctDNA provides comparable results with numerous diagnostic advantages for precision oncology.

STATEMENT OF SIGNIFICANCEThis study provides key insights into the dynamics of nucleosome positioning and gene regulation associated with cancer phenotypes that can be ascertained from ctDNA. The new methods established for phenotype classification extend the utility of ctDNA beyond assessments of DNA alterations with important implications for molecular diagnostics and precision oncology.
]]></description>
<dc:creator>De Sarkar, N.</dc:creator>
<dc:creator>Patton, R. D.</dc:creator>
<dc:creator>Doebley, A.-L.</dc:creator>
<dc:creator>Hanratty, B.</dc:creator>
<dc:creator>Kreitzman, A. J.</dc:creator>
<dc:creator>Sarthy, J. F.</dc:creator>
<dc:creator>Ko, M.</dc:creator>
<dc:creator>Adil, M.</dc:creator>
<dc:creator>Brahma, S.</dc:creator>
<dc:creator>Meers, M. P.</dc:creator>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Ang, L. S.</dc:creator>
<dc:creator>Coleman, I.</dc:creator>
<dc:creator>Bose, A.</dc:creator>
<dc:creator>Dumpit, R. F.</dc:creator>
<dc:creator>Lucas, J. M.</dc:creator>
<dc:creator>Nunez, T. A.</dc:creator>
<dc:creator>Nguyen, H. M.</dc:creator>
<dc:creator>McClure, H. M.</dc:creator>
<dc:creator>Pritchard, C. C.</dc:creator>
<dc:creator>Schweizer, M. T.</dc:creator>
<dc:creator>Morrissey, C.</dc:creator>
<dc:creator>Choudhury, A. D.</dc:creator>
<dc:creator>Baca, S. C.</dc:creator>
<dc:creator>Berchuck, J. E.</dc:creator>
<dc:creator>Freedman, M. L.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Montgomery, B.</dc:creator>
<dc:creator>Corey, E.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:date>2022-06-25</dc:date>
<dc:identifier>doi:10.1101/2022.06.21.496879</dc:identifier>
<dc:title><![CDATA[Nucleosome patterns in circulating tumor DNA reveal transcriptional regulation of advanced prostate cancer phenotypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.26.497647v1?rss=1">
<title>
<![CDATA[
Cross-Protective Antibodies Against Common Endemic Respiratory Viruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.26.497647v1?rss=1</link>
<description><![CDATA[
Respiratory syncytial virus (RSV), human metapneumovirus (HMPV), and human parainfluenza virus types one (HPIV1) and three (HPIV3) are a major cause of death, morbidity, and health care costs worldwide, and they can exact a significant toll on immunocompromised patients, the elderly, and those with underlying lung disease. There is an unmet medical need for safe and effective medications for many of the viruses responsible for common respiratory viral infections in vulnerable patient populations. While a protective monoclonal antibody exists for RSV, clinical use is limited to high-risk infant populations. Here, we present the discovery, in vitro characterization, and in vivo efficacy testing of two cross-neutralizing monoclonal antibodies, one targeting both HPIV3 and HPIV1 and the other targeting both RSV and HMPV. The 3x1 antibody is capable of targeting multiple parainfluenza viruses; the MxR antibody shares features with other previously reported monoclonal antibodies that are capable of neutralizing both RSV and HMPV. We obtained structures using cryo-electron microscopy of these antibodies in complex with their antigens to 3.62 [A] resolution for 3x1:HPIV3 and to 2.24 [A] for MxR:RSV, providing a structural basis to corroborate our in vitro characterization of binding and neutralization. Together, a cocktail of 3x1 and MxR could have clinical utility in providing broad protection against four of the respiratory viruses that cause significant morbidity and mortality in at-risk individuals.
]]></description>
<dc:creator>Caban, M.</dc:creator>
<dc:creator>Rodarte, J.</dc:creator>
<dc:creator>Bibby, M.</dc:creator>
<dc:creator>Gray, M. D.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>Boonyaratanakornkit, J.</dc:creator>
<dc:date>2022-06-26</dc:date>
<dc:identifier>doi:10.1101/2022.06.26.497647</dc:identifier>
<dc:title><![CDATA[Cross-Protective Antibodies Against Common Endemic Respiratory Viruses]]></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/10.1101/2022.07.06.498858v1?rss=1">
<title>
<![CDATA[
XDec Simplex Map of Breast Cancer Cell States Enables Precise Modeling and Targeting of Breast Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.06.498858v1?rss=1</link>
<description><![CDATA[
The characterization of cancer cell states within the tumor microenvironment is a key to understanding tumor biology and an important step toward the development of precision therapies. To reconstruct this information from bulk RNA-seq profiles, we developed the XDec Simplex Mapping (XDec-SM) approach, a reference-optional deconvolution method that leverages single-cell information, when such information is available, to map tumors and the states of constituent cells onto a biologically interpretable, low-dimensional space. When applied to breast tumors in The Cancer Genome Atlas (TCGA), XDec-SM infers the identity of constituent cell types and their proportions. XDec-SM also infers cancer cells states within individual tumors that associate with DNA methylation patterns, driver somatic mutations, pathway activation and metabolic coupling between stromal and breast cancer cells. By projecting tumors, cancer cell lines, and PDX models onto the same map, we identify both in vitro and in vivo models with matching cancer cell states. Map position is also predictive of therapy response, thus opening the prospects for precision therapy informed by experiments in model systems matched to tumors in vivo by cancer cell state.
]]></description>
<dc:creator>Murillo, O. D.</dc:creator>
<dc:creator>Petrosyan, V.</dc:creator>
<dc:creator>LaPlante, E. L.</dc:creator>
<dc:creator>Dobrolecki, L. E.</dc:creator>
<dc:creator>Lewis, M. T.</dc:creator>
<dc:creator>Milosavljevic, A.</dc:creator>
<dc:date>2022-07-07</dc:date>
<dc:identifier>doi:10.1101/2022.07.06.498858</dc:identifier>
<dc:title><![CDATA[XDec Simplex Map of Breast Cancer Cell States Enables Precise Modeling and Targeting of Breast Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.08.499199v1?rss=1">
<title>
<![CDATA[
Transient T cell expansion, activation, and proliferation in therapeutically vaccinated SIV+ macaques treated with N-803 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.08.499199v1?rss=1</link>
<description><![CDATA[
Vaccine strategies aimed at eliciting HIV-specific CD8+ T cells are one major target of interest in HIV functional cure strategies. We hypothesized that CD8+ T cells elicited by therapeutic vaccination during antiretroviral therapy (ART) would be recalled and boosted by treatment with the IL-15 superagonist N-803 after ART discontinuation. We intravenously immunized four SIV+ Mauritian cynomolgus macaques (MCM) receiving ART with vesicular stomatitis virus (VSV), modified vaccinia virus Ankara strain (MVA), and recombinant adenovirus serotype 5 (rAd-5) vectors all expressing SIVmac239 Gag. Immediately after ART cessation, these animals received three doses of N-803. Four control animals received no vaccines or N-803. The vaccine regimen generated a high magnitude of Gag-specific CD8+ T cells that were proliferative and biased toward an effector memory phenotype. We then compared cells elicited by vaccination (Gag-specific) to cells elicited by SIV infection and unaffected by vaccination (Nef-specific). We found that N-803 treatment enhanced both the frequencies of bulk and proliferating antigen-specific CD8+ T cells elicited by vaccination and the antigen-specific CD8+ T cells elicited by SIV infection. In sum, we demonstrate that a therapeutic heterologous prime-boost-boost (HPBB) vaccine can elicit antigen-specific effector memory CD8+ T cells that are boosted by N-803.
]]></description>
<dc:creator>Harwood, O. E.</dc:creator>
<dc:creator>Balgeman, A. J.</dc:creator>
<dc:creator>Weaver, A. J.</dc:creator>
<dc:creator>Ellis-Connell, A. L.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Erickson, K. N.</dc:creator>
<dc:creator>Matschke, L. M.</dc:creator>
<dc:creator>Golfinos, A. E.</dc:creator>
<dc:creator>Vezys, V.</dc:creator>
<dc:creator>Skinner, P. J.</dc:creator>
<dc:creator>Safrit, J. T.</dc:creator>
<dc:creator>Edlefsen, P. T.</dc:creator>
<dc:creator>Reynolds, M. R.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:creator>O'Connor, S. L.</dc:creator>
<dc:date>2022-07-08</dc:date>
<dc:identifier>doi:10.1101/2022.07.08.499199</dc:identifier>
<dc:title><![CDATA[Transient T cell expansion, activation, and proliferation in therapeutically vaccinated SIV+ macaques treated with N-803]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.25.501144v1?rss=1">
<title>
<![CDATA[
Extrachromosomal DNA in the cancerous transformation of Barrett's esophagus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.25.501144v1?rss=1</link>
<description><![CDATA[
BACKGROUNDOncogenes are commonly amplified on extrachromosomal DNA (ecDNA) contributing to poor outcomes for patients. Currently, the chronology of ecDNA development is not known. We studied the origination and evolution of ecDNA in patients with Barretts esophagus (BE) who progressed to esophageal adenocarcinoma (EAC).

METHODSWe analyzed whole-genome sequencing (WGS) data from a BE surveillance cohort and EAC patients at Cambridge University UK (n=206 patients). We also analyzed WGS data from biopsies taken at two time points from multiple sites in the esophagus from 80 patients enrolled in a case-control study at the Fred Hutchinson Cancer Center (FHCC) - 40 BE patients who progressed to EAC and 40 who did not.

RESULTSecDNA was detected in 24% and 43% of BE patients with BE-associated early and late-stage EAC, respectively, in the Cambridge cross-sectional cohort. ecDNA was found in 33% of all FHCC BE patients who developed cancer, either prior to, or at EAC diagnosis. ecDNA was strongly associated with patients who developed cancer, in contrast with FHCC BE patients who did not progress (odds ratio, 18.8, CI - 2.3-152, p=3.3x10-4). ecDNAs were enriched for oncogenes and immunomodulatory genes and could be detected early in the transition from high-grade dysplasia to cancer and increased in copy number and complexity over time.

CONCLUSIONSecDNAs can develop before a diagnosis of cancer in BE patients and are strongly selected for during the evolution to EAC. ecDNAs promote diverse oncogene and immunomodulatory gene amplification during EAC development and progression.
]]></description>
<dc:creator>Luebeck, J.</dc:creator>
<dc:creator>Ng, A. W. T.</dc:creator>
<dc:creator>Galipeau, P. C.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Sanchez, C. A.</dc:creator>
<dc:creator>Katz-Summercorn, A.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Jammula, S.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Lippman, S. M.</dc:creator>
<dc:creator>Verhaak, R. G. W.</dc:creator>
<dc:creator>Maley, C. C.</dc:creator>
<dc:creator>Alexandrov, L. B.</dc:creator>
<dc:creator>Reid, B. J.</dc:creator>
<dc:creator>Fitzgerald, R. C.</dc:creator>
<dc:creator>Paulson, T. G.</dc:creator>
<dc:creator>Chang, H. Y.</dc:creator>
<dc:creator>Wu, S.</dc:creator>
<dc:creator>Bafna, V.</dc:creator>
<dc:creator>Mischel, P. S.</dc:creator>
<dc:date>2022-07-25</dc:date>
<dc:identifier>doi:10.1101/2022.07.25.501144</dc:identifier>
<dc:title><![CDATA[Extrachromosomal DNA in the cancerous transformation of Barrett's esophagus]]></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/10.1101/2022.08.04.502764v1?rss=1">
<title>
<![CDATA[
Deep molecular, cellular and temporal phenotyping of developmental perturbations at whole organism scale 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.04.502764v1?rss=1</link>
<description><![CDATA[
The maturation of single cell transcriptomic technologies has facilitated the generation of comprehensive cellular atlases from whole embryos. A majority of this data, however, has been collected from wild type embryos without an appreciation for latent variation present in development. Here we present single cell transcriptomic data from 1812 individually resolved developing zebrafish embryos, encompassing 19 time points, 23 genetic perturbations, and totaling 3.2M cells. The high degree of replication in our study (8 or more embryos per condition) allows us to estimate the variance in cell type abundance organism-wide and to detect perturbation-dependent deviance in cell type composition relative to wild type embryos. Our approach is sensitive to rare cell types, resolving developmental trajectories and genetic dependencies in the cranial ganglia neurons, a cell population that comprises less than 1% of the embryo. Additionally, time-series profiling of individual mutants identified a group of brachyury-independent cells with strikingly similar transcriptomes to notochord sheath cells, leading to new hypotheses about the origins of the skull. We anticipate that standardized collection of high-resolution, organism-scale single cell data from large numbers of individual embryos will enable mapping the genetic dependencies of zebrafish cell types, while also addressing long-standing challenges in developmental genetics, including the cellular and transcriptional plasticity underlying phenotypic diversity across individuals.
]]></description>
<dc:creator>Saunders, L. M.</dc:creator>
<dc:creator>Srivatsan, S. R.</dc:creator>
<dc:creator>Duran, M.</dc:creator>
<dc:creator>Dorrity, M. W.</dc:creator>
<dc:creator>Ewing, B.</dc:creator>
<dc:creator>Linbo, T.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Raible, D.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:creator>Kimelman, D.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:date>2022-08-05</dc:date>
<dc:identifier>doi:10.1101/2022.08.04.502764</dc:identifier>
<dc:title><![CDATA[Deep molecular, cellular and temporal phenotyping of developmental perturbations at whole organism scale]]></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/10.1101/2022.08.05.502978v1?rss=1">
<title>
<![CDATA[
State-dependent evolutionary models reveal modes of solid tumor growth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.05.502978v1?rss=1</link>
<description><![CDATA[
Spatial properties of tumor growth have profound implications for cancer progression, therapeutic resistance and metastasis. Yet, how spatial position governs tumor cell division remains difficult to evaluate in clinical tumors. Here, we demonstrate that elevated cellular growth rates on the tumor periphery leave characteristic patterns in the genomes of cells sampled from different parts of a tumor, which become evident when they are used to construct a tumor phylogenetic tree. Namely, rapidly-dividing peripheral lineages branch more extensively and acquire more mutations than slower-dividing lineages in the tumor center. We develop a Bayesian state-dependent evolutionary phylodynamic model (SDevo) that quantifies these patterns to infer the differential cell division rates between peripheral and central cells jointly from the branching and mutational patterns of single-time point, multi-region sequencing data. We validate this approach on simulated tumors by demonstrating its ability to accurately infer spatially-varying birth rates under a range of growth conditions and sampling strategies. We then show that SDevo outperforms state-of-the-art, non-cancer multi-state phylodynamic methods which ignore differential mutational acquisition. Finally, we apply SDevo to multi-region sequencing data from clinical hepatocellular carcinomas and find evidence that cells on the tumor edge divide 3-6x faster than those in the center. As multi-region and single-cell sequencing increase in resolution and availability, we anticipate that SDevo will be useful in interrogating spatial restrictions on tumor growth and could be extended to model non-spatial factors that influence tumor progression, including hypoxia and immune infiltration.
]]></description>
<dc:creator>Lewinsohn, M. A.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Müller, N. F.</dc:creator>
<dc:creator>Feder, A. F.</dc:creator>
<dc:date>2022-08-06</dc:date>
<dc:identifier>doi:10.1101/2022.08.05.502978</dc:identifier>
<dc:title><![CDATA[State-dependent evolutionary models reveal modes of solid tumor growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.23.504195v1?rss=1">
<title>
<![CDATA[
A modular CRISPR screen identifies individual and combination pathways contributing to HIV-1 latency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.23.504195v1?rss=1</link>
<description><![CDATA[
Transcriptional silencing of latent HIV-1 proviruses entails complex and overlapping mechanisms that pose a major barrier to in vivo elimination of HIV-1. We developed a new latency CRISPR screening strategy, called Latency HIV-CRISPR, which uses the packaging of guideRNA-encoding lentiviral vector genomes into the supernatant of budding virions as a direct readout of factors involved in the maintenance of HIV-1 latency. We developed a custom guideRNA library targeting epigenetic regulatory genes and paired the screen with and without a latency reversal agent - AZD5582, an activator of the non-canonical NF{kappa}B pathway - to examine a combination of mechanisms controlling HIV-1 latency. A component of the Nucleosome Acetyltransferase of H4 histone acetylation (NuA4 HAT) complex, ING3, acts in concert with AZD5582 to activate proviruses in J-Lat cell lines and in a primary CD4+ T cell model of HIV-1 latency. We found that the knockout of ING3 reduces acetylation of the H4 histone tail and BRD4 occupancy on the HIV-1 LTR, and only in the combination of ING3 knockout with the activation of non-canonical NF{kappa}B via AZD5582 is there dramatic increase in initiation and elongation of RNA Polymerase II on the HIV-1 provirus in a manner that is nearly unique among all cellular promoters.
]]></description>
<dc:creator>Hsieh, E.</dc:creator>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Paddison, P. J.</dc:creator>
<dc:creator>Browne, E. P.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Ohainle, M.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2022-08-23</dc:date>
<dc:identifier>doi:10.1101/2022.08.23.504195</dc:identifier>
<dc:title><![CDATA[A modular CRISPR screen identifies individual and combination pathways contributing to HIV-1 latency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.24.505164v1?rss=1">
<title>
<![CDATA[
Safety switch optimization enhances antibody-mediated elimination of CAR T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.24.505164v1?rss=1</link>
<description><![CDATA[
Activation of a conditional safety switch has the potential to reverse serious toxicities arising from the administration of engineered cellular therapies, including chimeric antigen receptor (CAR) T cells. The functionally inert, non-immunogenic cell surface marker derived from human epidermal growth factor receptor (EGFRt) is a promising safety switch that has been used in multiple clinical constructs and can be targeted by cetuximab, a clinically available monoclonal antibody. However, this approach requires high and persistent cell surface expression of EGFRt to ensure that antibody mediated depletion of engineered cells is rapid and complete. Here we show that incorporating a short juxtamembrane sequence into the EGFRt polypeptide enhances its expression on the surface of T cells and their susceptibility to antibody-dependent cellular cytotoxicity (ADCC). Incorporating this optimized variant (EGFRopt) into bicistronic and tricistronic CAR designs results in more rapid in vivo elimination of CAR T cells and robust termination of their effector activity compared to EGFRt. These studies establish EGFRopt as a superior safety switch for the development of next-generation cell-based therapeutics.
]]></description>
<dc:creator>Shabaneh, T. B.</dc:creator>
<dc:creator>Moffett, H. F.</dc:creator>
<dc:creator>Stull, S. M.</dc:creator>
<dc:creator>Derezes, T.</dc:creator>
<dc:creator>Tait, L. J.</dc:creator>
<dc:creator>Park, S.</dc:creator>
<dc:creator>Riddell, S. R.</dc:creator>
<dc:creator>Lajoie, M. J.</dc:creator>
<dc:date>2022-08-24</dc:date>
<dc:identifier>doi:10.1101/2022.08.24.505164</dc:identifier>
<dc:title><![CDATA[Safety switch optimization enhances antibody-mediated elimination of CAR T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.29.505713v1?rss=1">
<title>
<![CDATA[
Receptor binding domain (RBD) antibodies contribute more to SARS-CoV-2 neutralization when target cells express high levels of ACE2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.29.505713v1?rss=1</link>
<description><![CDATA[
Neutralization assays are experimental surrogates for the effectiveness of infection- or vaccine-elicited polyclonal antibodies and therapeutic monoclonal antibodies targeting SARS-CoV-2. However, the measured neutralization can depend on details of the experimental assay. Here we systematically assess how ACE2 expression in target cells affects neutralization by antibodies to different spike epitopes in lentivirus pseudovirus neutralization assays. For high ACE2-expressing target cells, receptor binding domain (RBD) antibodies account for nearly all neutralizing activity in polyclonal human sera. But for lower ACE2-expressing target cells, antibodies targeting regions outside the RBD make a larger (although still modest) contribution to serum neutralization. These serum-level results are mirrored for monoclonal antibodies: N-terminal domain (NTD) antibodies and RBD antibodies that do not compete for ACE2 binding incompletely neutralize on high ACE2-expressing target cells, but completely neutralize on cells with lower ACE2 expression. Our results show that ACE2 expression level in the target cells is an important experimental variable, and that high ACE2 expression emphasizes the role of a subset of RBD-directed antibodies.
]]></description>
<dc:creator>Farrell, A. G.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Eguia, R.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Francko, N. M.</dc:creator>
<dc:creator>Logue, J.</dc:creator>
<dc:creator>Carreno, J. M.</dc:creator>
<dc:creator>Abbad, A.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Matreyek, K. A.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2022-08-30</dc:date>
<dc:identifier>doi:10.1101/2022.08.29.505713</dc:identifier>
<dc:title><![CDATA[Receptor binding domain (RBD) antibodies contribute more to SARS-CoV-2 neutralization when target cells express high levels of ACE2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.30.505828v1?rss=1">
<title>
<![CDATA[
Influenza virus transcription and progeny production are poorly correlated in single cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.30.505828v1?rss=1</link>
<description><![CDATA[
The ultimate success of a viral infection at the cellular level is determined by the number of progeny virions produced. However, most single-cell studies of infection quantify the expression of viral transcripts and proteins, rather than the amount of progeny virions released from infected cells. Here we overcome this limitation by simultaneously measuring transcription and progeny production from single influenza-virus-infected cells by embedding nucleotide barcodes in the viral genome. We find that viral transcription and progeny production are poorly correlated in single cells. The cells that transcribe the most viral mRNA do not produce the most viral progeny, and often represent aberrant infections that fail to express the influenza NS gene. However, only some of the discrepancy between transcription and progeny production can be explained by viral gene absence or mutations: there is also a wide range of progeny production among cells infected by complete unmutated virions. Overall, our results show that viral transcription is a relatively poor predictor of an infected cells contribution to the progeny population.
]]></description>
<dc:creator>Bacsik, D. J.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Butler, A.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Heaton, N. S.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2022-08-30</dc:date>
<dc:identifier>doi:10.1101/2022.08.30.505828</dc:identifier>
<dc:title><![CDATA[Influenza virus transcription and progeny production are poorly correlated in single cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.11.507488v1?rss=1">
<title>
<![CDATA[
Fanconi anemia-isogenic head and neck cancer cell line pairs - a basic and translational science resource 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.11.507488v1?rss=1</link>
<description><![CDATA[
Fanconi anemia (FA) is a heritable malformation, bone marrow failure and cancer predisposition syndrome that confers an exceptionally high risk of developing carcinomas arising in squamous mucosal epithelia lining the mouth, proximal esophagus, vulva and anus. The origin of these cancers is not understood, and no effective way has been identified to prevent or delay their appearance. FA-associated carcinomas are also therapeutically challenging, as they may be multi-focal and stage-advanced at diagnosis making surgical control challenging. Moreover, individuals with FA have systemic DNA damage hypersensitivity and thus an elevated risk of toxicity when treated with standard-of-care therapies such as DNA cross-linking drugs and ionizing radiation.

We developed the Fanconi Anemia Cancer Cell Line Resource (FA-CCLR) in order to foster new research on the origins, treatment, and prevention of FA-associated cancers. The FA-CCLR consists of FANC-isogenic head and neck squamous cell carcinoma (HNSCC) cell line pairs from cancers arising in individuals with FA, or newly engineered from sporadic HNSCC cell lines. Molecular, cellular, and biochemical analyses were used to demonstrate the causal dependence of key FA-associated phenotypes on FANC genotype, expression and pathway activity. These FANC-isogenic cell line pairs are available to academic and non-profit investigators, with ordering information available at the  Fanconi Anemia Research Materials Resource and Repository at Oregon Health & Sciences University, Portland OR.

SignificanceWe have generated new isogenic cancer cell line models to investigate the origins, treatment and prevention of Fanconi anemia-associated squamous carcinomas that target the oral mucosa, proximal esophagus, and anogenital region.
]]></description>
<dc:creator>Nguyen, H. T.</dc:creator>
<dc:creator>Tang, W.</dc:creator>
<dc:creator>Webster, A. L. H.</dc:creator>
<dc:creator>Whiteaker, J. R.</dc:creator>
<dc:creator>Chandler, C. M.</dc:creator>
<dc:creator>Errazquin, R.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:creator>Jonlin, E.</dc:creator>
<dc:creator>Hoskins, E. E.</dc:creator>
<dc:creator>Chen, E. Y.</dc:creator>
<dc:creator>Fritzke, M.</dc:creator>
<dc:creator>Paulovich, A. G.</dc:creator>
<dc:creator>Wells, S. I.</dc:creator>
<dc:creator>Roohollahi, K.</dc:creator>
<dc:creator>Dorsman, J.</dc:creator>
<dc:creator>Brakenhoff, R.</dc:creator>
<dc:creator>Garcia-Escudero, R.</dc:creator>
<dc:creator>Smogorzewska, A.</dc:creator>
<dc:creator>Wakefield, L.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Monnat, R. J.</dc:creator>
<dc:date>2022-09-13</dc:date>
<dc:identifier>doi:10.1101/2022.09.11.507488</dc:identifier>
<dc:title><![CDATA[Fanconi anemia-isogenic head and neck cancer cell line pairs - a basic and translational science resource]]></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/10.1101/2022.09.17.508366v1?rss=1">
<title>
<![CDATA[
A biophysical model of viral escape from polyclonal antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.17.508366v1?rss=1</link>
<description><![CDATA[
A challenge in studying viral immune escape is determining how mutations combine to escape polyclonal antibodies, which can potentially target multiple distinct viral epitopes. Here we introduce a biophysical model of this process that partitions the total polyclonal antibody activity by epitope, and then quantifies how each viral mutation affects the antibody activity against each epitope. We develop software that can use deep mutational scanning data to infer these properties for polyclonal antibody mixtures. We validate this software using a computationally simulated deep mutational scanning experiment, and demonstrate that it enables the prediction of escape by arbitrary combinations of mutations. The software described in this paper is available at https://jbloomlab.github.io/polyclonal.
]]></description>
<dc:creator>Yu, T. C.</dc:creator>
<dc:creator>Thornton, Z. T.</dc:creator>
<dc:creator>Hannon, W. H.</dc:creator>
<dc:creator>DeWitt, W. S.</dc:creator>
<dc:creator>Radford, C. E.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2022-09-17</dc:date>
<dc:identifier>doi:10.1101/2022.09.17.508366</dc:identifier>
<dc:title><![CDATA[A biophysical model of viral escape from polyclonal antibodies]]></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/10.1101/2022.09.20.508745v1?rss=1">
<title>
<![CDATA[
Deep mutational scans for ACE2 binding, RBD expression, and antibody escape in the SARS-CoV-2 Omicron BA.1 and BA.2 receptor-binding domains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.20.508745v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 continues to acquire mutations in the spike receptor-binding domain (RBD) that impact ACE2 receptor binding, folding stability, and antibody recognition. Deep mutational scanning prospectively characterizes the impacts of mutations on these biochemical properties, enabling rapid assessment of new mutations seen during viral surveillance. However, the effects of mutations can change as the virus evolves, requiring updated deep mutational scans. We determined the impacts of all amino acid mutations in the Omicron BA.1 and BA.2 RBDs on ACE2-binding affinity, RBD folding, and escape from binding by the LY-CoV1404 (bebtelovimab) monoclonal antibody. The effects of some mutations in Omicron RBDs differ from those measured in the ancestral Wuhan-Hu-1 background. These epistatic shifts largely resemble those previously seen in the Beta variant due to the convergent epistatically modifying N501Y substitution. However, Omicron variants show additional lineage-specific shifts, including examples of the epistatic phenomenon of entrenchment that causes the Q498R and N501Y substitutions present in Omicron to be more favorable in that background than in earlier viral strains. In contrast, the Omicron substitution Q493R exhibits no sign of entrenchment, with the derived state, R493, being as unfavorable for ACE2 binding in Omicron RBDs as in Wuhan-Hu-1. Likely for this reason, the R493Q reversion has occurred in Omicron sub-variants including BA.4/BA.5 and BA.2.75, where the affinity buffer from R493Q reversion may potentiate concurrent antigenic change. Consistent with prior studies, we find that Omicron RBDs have reduced expression, and identify candidate stabilizing mutations that ameliorate this deficit. Last, our maps highlight a broadening of the sites of escape from LY-CoV1404 antibody binding in BA.1 and BA.2 compared to the ancestral Wuhan-Hu-1 background. These BA.1 and BA.2 deep mutational scanning datasets identify shifts in the RBD mutational landscape and inform ongoing efforts in viral surveillance.

Author SummarySARS-CoV-2 evolves in part through mutations in its spike receptor-binding domain. As these mutations accumulate in evolved variants, they shape the future evolutionary potential of the virus through the phenomenon of epistasis. We characterized the functional impacts of mutations in the Omicron BA.1 and BA.2 receptor-binding domains on ACE2 receptor binding, protein folding, and recognition by the clinical LY-CoV1404 antibody. We then compared the measurements to prior data for earlier variants. These comparisons identify patterns of epistasis that may alter future patterns of Omicron evolution, such as turnover in the availability of specific affinity-enhancing mutations and an expansion in the number of paths of antibody escape from a key monoclonal antibody used for therapeutic treatment of COVID-19. This work informs continued efforts in viral surveillance and forecasting.
]]></description>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Stewart, C. M.</dc:creator>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>Hannon, W. W.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2022-09-20</dc:date>
<dc:identifier>doi:10.1101/2022.09.20.508745</dc:identifier>
<dc:title><![CDATA[Deep mutational scans for ACE2 binding, RBD expression, and antibody escape in the SARS-CoV-2 Omicron BA.1 and BA.2 receptor-binding domains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.04.510817v1?rss=1">
<title>
<![CDATA[
A human-specific motif facilitates CARD8 inflammasome activation after HIV-1 infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.04.510817v1?rss=1</link>
<description><![CDATA[
Inflammasomes are cytosolic innate immune complexes that play a critical role in host defense against pathogens but can also contribute to inflammatory pathogenesis. Here, we find that the human inflammasome-forming sensor CARD8 senses HIV-1 infection via site-specific cleavage of the CARD8 N-terminus by the HIV protease (HIV-1PR). HIV-1PR cleavage of CARD8 induces pyroptotic cell death and the release of pro-inflammatory cytokines from infected cells, processes that we find are dependent on Toll-like receptor stimulation prior to viral infection. Our evolutionary analyses reveal that the HIV-1PR cleavage site in CARD8 is unique to humans, and that chimpanzee CARD8 does not recognize proteases from HIV or simian immunodeficiency viruses from chimpanzees (SIVcpz). In contrast, SIVcpz does cleave human CARD8, suggesting that SIVcpz was poised to activate the human CARD8 inflammasome prior to its cross-species transmission into humans and implicating the CARD8 inflammasome as a potential driver of HIV pathogenesis.
]]></description>
<dc:creator>Kulsuptrakul, J.</dc:creator>
<dc:creator>Turcotte, E.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Mitchell, P.</dc:creator>
<dc:date>2022-10-04</dc:date>
<dc:identifier>doi:10.1101/2022.10.04.510817</dc:identifier>
<dc:title><![CDATA[A human-specific motif facilitates CARD8 inflammasome activation after HIV-1 infection]]></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/10.1101/2022.10.06.511235v1?rss=1">
<title>
<![CDATA[
An optimized gene knock-out strategy for human CD34+ hematopoietic stem and progenitor cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.06.511235v1?rss=1</link>
<description><![CDATA[
Human CD34+ hematopoietic stem and progenitor cells (HSPCs) are a standard source of cells for clinical HSC transplantations as well as experimental xenotransplantation to generate "humanized mice". To further extend the range of applications of these humanized mice, we developed a protocol to efficiently edit the genomes of human CD34+ HSPCs before transplantation. In the past, manipulating HSPCs has been complicated by the fact that they are inherently difficult to transduce with lentivectors, and rapidly lose their stemness and engraftment potential during in vitro culture. However, with optimized nucleofection of sgRNA:Cas9 ribonucleoprotein complexes, we are now able to edit a candidate gene in CD34+ HSPCs with almost 100% efficiency, and without affecting their potential for engraftment and multilineage differentiation in mice. The result is a humanized mouse from which we knocked out a gene of interest from their human immune system.
]]></description>
<dc:creator>Kuppers, D.</dc:creator>
<dc:creator>Linton, J.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:creator>Rongvaux, A.</dc:creator>
<dc:date>2022-10-07</dc:date>
<dc:identifier>doi:10.1101/2022.10.06.511235</dc:identifier>
<dc:title><![CDATA[An optimized gene knock-out strategy for human CD34+ hematopoietic stem and progenitor cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.12.511825v1?rss=1">
<title>
<![CDATA[
MR1-restricted T cell clonotypes are associated with 'resistance' to M.tuberculosis infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.12.511825v1?rss=1</link>
<description><![CDATA[
T cells are required for a protective immune response against the human adapted pathogen Mycobacterium tuberculosis (M.tb). We recently described a cohort of Ugandan household contacts of tuberculosis cases that appear to  resist M.tb infection (RSTRs) and showed that these individuals harbor IFN-{gamma} independent T cell responses to M.tb-specific peptide antigens. However, T cells also recognize non-protein antigens via antigen presenting systems that are independent of genetic background, leading to their designation as donor-unrestricted T (DURT) cells. We used combinatorial tetramer staining and multi-parameter flow cytometry to comprehensively characterize the association between DURTs and  resistance to M.tb infection. We did not observe a difference in peripheral blood frequencies of invariant natural killer T (iNKT) cells, germline encoded mycolyl-reactive (GEM) T cells, or {gamma}{delta} T cells between RSTRs and matched controls with latent M.tb infection (LTBIs). However, we did observe a 1.65-fold increase in frequency of circulating MR1-restricted T (MR1T) cells among RSTRs in comparison with LTBI (p=0.03). Multi-modal single cell RNA-sequencing of 18,251 MR1T cells sorted from a subset of donors revealed 5150 clonotypes that expressed a common transcriptional program, the majority of which were private. Deep sequencing of the TCR- repertoire revealed several DURT clonotypes that were expanded among RSTRs, including at least two MR1T clonotypes. Taken together, our data reveal unexpected donor-specific diversity in the TCR repertoire of human MR1T cells as well as associations between MR1 clonotypes and  resistance to M.tb infection.
]]></description>
<dc:creator>Cross, D. L.</dc:creator>
<dc:creator>Layton, E. D.</dc:creator>
<dc:creator>Yu, K. K.</dc:creator>
<dc:creator>Smith, M. T.</dc:creator>
<dc:creator>Aguilar, M. S.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Mayanja-Kizza, H.</dc:creator>
<dc:creator>Stein, C. M.</dc:creator>
<dc:creator>Boom, W. H.</dc:creator>
<dc:creator>Hawn, T. R.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Newell, E.</dc:creator>
<dc:creator>Seshadri, C.</dc:creator>
<dc:date>2022-10-13</dc:date>
<dc:identifier>doi:10.1101/2022.10.12.511825</dc:identifier>
<dc:title><![CDATA[MR1-restricted T cell clonotypes are associated with 'resistance' to M.tuberculosis infection]]></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/10.1101/2022.10.13.512056v1?rss=1">
<title>
<![CDATA[
A pseudovirus system enables deep mutational scanning of the full SARS-CoV-2 spike 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.13.512056v1?rss=1</link>
<description><![CDATA[
A major challenge in understanding SARS-CoV-2 evolution is interpreting the antigenic and functional effects of emerging mutations in the viral spike protein. Here we describe a new deep mutational scanning platform based on non-replicative pseudotyped lentiviruses that directly quantifies how large numbers of spike mutations impact antibody neutralization and pseudovirus infection. We demonstrate this new platform by making libraries of the Omicron BA.1 and Delta spikes. These libraries each contain ~7000 distinct amino-acid mutations in the context of up to ~135,000 unique mutation combinations. We use these libraries to map escape mutations from neutralizing antibodies targeting the receptor binding domain, N-terminal domain, and S2 subunit of spike. Overall, this work establishes a high-throughput and safe approach to measure how ~105 combinations of mutations affect antibody neutralization and spike-mediated infection. Notably, the platform described here can be extended to the entry proteins of many other viruses.
]]></description>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Radford, C. E.</dc:creator>
<dc:creator>Farrell, A. G.</dc:creator>
<dc:creator>Yu, T. C.</dc:creator>
<dc:creator>Hannon, W. W.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Andrabi, R.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Ho, D. D.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2022-10-13</dc:date>
<dc:identifier>doi:10.1101/2022.10.13.512056</dc:identifier>
<dc:title><![CDATA[A pseudovirus system enables deep mutational scanning of the full SARS-CoV-2 spike]]></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/10.1101/2022.10.21.513237v1?rss=1">
<title>
<![CDATA[
Computationally restoring the potency of a clinical antibody against SARS-CoV-2 Omicron subvariants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.21.513237v1?rss=1</link>
<description><![CDATA[
The COVID-19 pandemic underscored the promise of monoclonal antibody-based prophylactic and therapeutic drugs1-3, but also revealed how quickly viral escape can curtail effective options4, 5. With the emergence of the SARS-CoV-2 Omicron variant in late 2021, many clinically used antibody drug products lost potency, including EvusheldTM and its constituent, cilgavimab4, 6. Cilgavimab, like its progenitor COV2-2130, is a class 3 antibody that is compatible with other antibodies in combination4 and is challenging to replace with existing approaches. Rapidly modifying such high-value antibodies with a known clinical profile to restore efficacy against emerging variants is a compelling mitigation strategy. We sought to redesign COV2-2130 to rescue in vivo efficacy against Omicron BA.1 and BA.1.1 strains while maintaining efficacy against the contemporaneously dominant Delta variant. Here we show that our computationally redesigned antibody, 2130-1-0114-112, achieves this objective, simultaneously increases neutralization potency against Delta and many variants of concern that subsequently emerged, and provides protection in vivo against the strains tested, WA1/2020, BA.1.1, and BA.5. Deep mutational scanning of tens of thousands pseudovirus variants reveals 2130-1-0114-112 improves broad potency without incurring additional escape liabilities. Our results suggest that computational approaches can optimize an antibody to target multiple escape variants, while simultaneously enriching potency. Because our approach is computationally driven, not requiring experimental iterations or pre-existing binding data, it could enable rapid response strategies to address escape variants or pre-emptively mitigate escape vulnerabilities.
]]></description>
<dc:creator>Desautels, T. A.</dc:creator>
<dc:creator>Arrildt, K. T.</dc:creator>
<dc:creator>Zemla, A. T.</dc:creator>
<dc:creator>Lau, E. Y.</dc:creator>
<dc:creator>Zhu, F.</dc:creator>
<dc:creator>Ricci, D.</dc:creator>
<dc:creator>Cronin, S.</dc:creator>
<dc:creator>Zost, S.</dc:creator>
<dc:creator>Binshtein, E.</dc:creator>
<dc:creator>Scheaffer, S. M.</dc:creator>
<dc:creator>Engdahl, T. B.</dc:creator>
<dc:creator>Chen, E.</dc:creator>
<dc:creator>Goforth, J. W.</dc:creator>
<dc:creator>Vashchenko, D.</dc:creator>
<dc:creator>Nguyen, S.</dc:creator>
<dc:creator>Weilhammer, D. R.</dc:creator>
<dc:creator>Lo, J. K.-Y.</dc:creator>
<dc:creator>Rubinfeld, B.</dc:creator>
<dc:creator>Saada, E. A.</dc:creator>
<dc:creator>Weisenberger, T.</dc:creator>
<dc:creator>Lee, T.-H.</dc:creator>
<dc:creator>Whitener, B.</dc:creator>
<dc:creator>Case, J. B.</dc:creator>
<dc:creator>Ladd, A.</dc:creator>
<dc:creator>Silva, M. S.</dc:creator>
<dc:creator>Haluska, R. M.</dc:creator>
<dc:creator>Grzesiak, E. A.</dc:creator>
<dc:creator>Bates, T. W.</dc:creator>
<dc:creator>Petersen, B. K.</dc:creator>
<dc:creator>Thackray, L. B.</dc:creator>
<dc:creator>Segelke, B. W.</dc:creator>
<dc:creator>Lillo, A. M.</dc:creator>
<dc:creator>Sundaram, S.</dc:creator>
<dc:creator>Diamond, M. S.</dc:creator>
<dc:creator>Crowe, J. E.</dc:creator>
<dc:creator>Carnahan, R. H.</dc:creator>
<dc:creator>Faissol, D. M.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.21.513237</dc:identifier>
<dc:title><![CDATA[Computationally restoring the potency of a clinical antibody against SARS-CoV-2 Omicron subvariants]]></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/10.1101/2022.10.22.513040v1?rss=1">
<title>
<![CDATA[
DNA methylation restricts coordinated germline and neural fates in embryonic stem cell differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.22.513040v1?rss=1</link>
<description><![CDATA[
Somatic DNA methylation is established early during mammalian development, as embryonic cells transition from naive to primed pluripotency. This precedes the emergence of the three somatic germ layers, but also the segregation of the germline that undergoes genome-wide DNA demethylation after specification. While DNA methylation is essential for embryogenesis, the point at which it becomes critical during differentiation and whether all lineages equally depend on it is unclear. Using culture modeling of cellular transitions, we found that DNA methylation-free embryonic stem cells (ESCs) with a triple DNA methyltransferase knockout (TKO) normally progressed through the continuum of pluripotency states, but demonstrated skewed differentiation abilities towards neural versus other somatic lineages. More saliently, TKO ESCs were fully competent for establishing primordial germ cell-like cells (PGCLCs), even showing temporally extended and self-sustained capacity for the germline fate. By mapping chromatin states, we found that the neural and germline lineages are linked by a similar enhancer dynamics during priming, defined by common sets of methyl-sensitive transcription factors that fail to be decommissioned in absence of DNA methylation. We propose that DNA methylation controls the temporality of a coordinated neural-germline axis of preferred differentiation route during early development.
]]></description>
<dc:creator>Schulz, M.</dc:creator>
<dc:creator>Teissandier, A.</dc:creator>
<dc:creator>de la Mata-Santaella, E.</dc:creator>
<dc:creator>Armand, M.</dc:creator>
<dc:creator>Iranzo, J.</dc:creator>
<dc:creator>El Marjou, F.</dc:creator>
<dc:creator>Gestraud, P.</dc:creator>
<dc:creator>Walter, M.</dc:creator>
<dc:creator>Kinston, S.</dc:creator>
<dc:creator>Göttgens, B.</dc:creator>
<dc:creator>Greenberg, M. V. C.</dc:creator>
<dc:creator>Bourc'his, D.</dc:creator>
<dc:date>2022-10-22</dc:date>
<dc:identifier>doi:10.1101/2022.10.22.513040</dc:identifier>
<dc:title><![CDATA[DNA methylation restricts coordinated germline and neural fates in embryonic stem cell differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.24.513230v1?rss=1">
<title>
<![CDATA[
Validation of antigen tetramers for the detection of antigen-specific B cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.24.513230v1?rss=1</link>
<description><![CDATA[
The study of antigen-specific lymphocytes has been a key advancement in immunology the past few decades. The development of multimerized probes containing antigens, peptide:MHC complexes, or other ligands was one innovation allowing the direct study antigen-specific lymphocytes by flow cytometry. While these types of studies are now common and performed by thousands of laboratories, quality control and assessment of probe quality is often minimal. In fact, many of these types of probes are made in-house and protocols vary between labs. While peptide:MHC multimers can often be obtained from commercial sources or core facilities, few such services exist for antigen multimers. To ensure high quality and consistency with ligand probes, we have developed an easy and robust multiplexed approach using commercially available beads able to bind antibodies specific for the ligand of interest. Using this assay, we have sensitively assessed the performance of peptide:MHC and antigen tetramers and find considerable batch-to-batch variability in performance and stability over time. This assay can also reveal common production errors such as miscalculation of antigen concentration. Unexpectedly, probes including the fluorochrome allophycocyanin exhibited more rapid performance decline compared to probes including the fluorochrome R-phycoerythrin when both were stored at 4{degrees}C. This performance decline was reduced for most, but not all, batches when antigen tetramers were instead stored at -20{degrees}C in 50% glycerol. This work could set the stage for the development of standardized assays for all commonly used ligand probes to limit lab-to-lab technical variation, and experimental failure due to probe underperformance.
]]></description>
<dc:creator>Fitzpatrick, K. S.</dc:creator>
<dc:creator>Poljakov, K.</dc:creator>
<dc:creator>Bibby, M. G.</dc:creator>
<dc:creator>Remington, A. J.</dc:creator>
<dc:creator>Gray, M. D.</dc:creator>
<dc:creator>Boonyaratanakornkit, J.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.24.513230</dc:identifier>
<dc:title><![CDATA[Validation of antigen tetramers for the detection of antigen-specific B cells]]></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/10.1101/2022.10.25.513724v1?rss=1">
<title>
<![CDATA[
Multi-species analysis of inflammatory response elements reveals ancient and lineage-specific contributions of transposable elements to NF-κB binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.25.513724v1?rss=1</link>
<description><![CDATA[
Transposable elements (TEs) provide a source of transcription factor binding sites that can rewire conserved gene regulatory networks. NF-{kappa}B is an evolutionary conserved transcription factor complex primarily involved in innate immunity and inflammation. The extent to which TEs have contributed to NF-{kappa}B responses during mammalian evolution is not well established. Here we performed a multi-species analysis of TEs bound by the NF-{kappa}B subunit RELA (also known as p65) in response to the proinflammatory cytokine TNF. By comparing RELA ChIP-seq data from TNF-stimulated primary aortic endothelial cells isolated from human, mouse and cow, we found that 55 TE subfamilies were associated with RELA bound regions. These RELA-bound transposons possess active epigenetic features and reside near TNF-responsive genes. A prominent example of lineage-specific contribution of transposons comes from the bovine SINE subfamilies Bov-tA1/2/3 which collectively contributed over 14,000 RELA bound regions in cow. By comparing RELA binding data across species, we also found several examples of RELA motif-bearing TEs that colonized the genome prior to the divergence of the three species and contributed to species-specific RELA binding. For example, we found human RELA bound MER81 instances were enriched for the interferon gamma pathway and demonstrated that one RELA bound MER81 element can control the TNF-induced expression of Interferon Gamma Receptor 2 (IFNGR2). Using ancestral reconstructions, we found that RELA containing MER81 instances rapidly decayed during early primate evolution (> 50 million years ago (MYA)) before stabilizing since the separation of Old World monkeys (< 50 MYA). Taken together, our results suggest ancient and lineage-specific transposon subfamilies contributed to mammalian NF-{kappa}B regulatory networks.
]]></description>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Alizada, A.</dc:creator>
<dc:creator>Rathnakumar, K.</dc:creator>
<dc:creator>Khyzha, N.</dc:creator>
<dc:creator>Taylor, T.</dc:creator>
<dc:creator>Campitelli, L.</dc:creator>
<dc:creator>Patel, Z. M.</dc:creator>
<dc:creator>Antounians, L.</dc:creator>
<dc:creator>Hughes, T.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:creator>Mitchell, J. A.</dc:creator>
<dc:creator>Fish, J. E.</dc:creator>
<dc:creator>Wilson, M. D.</dc:creator>
<dc:date>2022-10-26</dc:date>
<dc:identifier>doi:10.1101/2022.10.25.513724</dc:identifier>
<dc:title><![CDATA[Multi-species analysis of inflammatory response elements reveals ancient and lineage-specific contributions of transposable elements to NF-κB binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.27.514108v1?rss=1">
<title>
<![CDATA[
Tracking the horizontal transfer of plasmids in Shigella sonnei and Shigella flexneri using phylogenetics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.27.514108v1?rss=1</link>
<description><![CDATA[
The  silent pandemic of antimicrobial resistance (AMR) represents a significant global public health threat. AMR genes in bacteria are often carried on mobile elements, such as plasmids. The horizontal movement of plasmids allows AMR genes and resistance to key therapeutics to disseminate in a population. However, the quantification of the movement of plasmids remains challenging with existing computational approaches. Here, we introduce a novel method that allows us to reconstruct and quantify the movement of plasmids in bacterial populations over time. To do so, we model chromosomal and plasmid DNA co-evolution using a joint coalescent and plasmid transfer process in a Bayesian phylogenetic network approach. This approach reconstructs differences in the evolutionary history of plasmids and chromosomes to reconstruct instances where plasmids likely move between bacterial lineages while accounting for parameter uncertainty. We apply this new approach to a five-year dataset of Shigella, exploring the plasmid transfer rates of five different plasmids with different AMR and virulence profiles. In doing so, we reconstruct the co-evolution of the large Shigella virulence plasmid with the chromosome DNA. We quantify higher plasmid transfer rates of three small plasmids that move between lineages of Shigella sonnei. Finally, we determine the recent dissemination of a multidrug-resistant plasmid between S. sonnei and S. flexneri lineages in multiple independent events and through steady growth in prevalence since 2010. This approach has a strong potential to improve our understanding of the evolutionary dynamics of AMR-carrying plasmids as they are introduced, circulate, and are maintained in bacterial populations.
]]></description>
<dc:creator>Müller, N. F.</dc:creator>
<dc:creator>Duchene, S.</dc:creator>
<dc:creator>Williamson, D. A.</dc:creator>
<dc:creator>Howden, B.</dc:creator>
<dc:creator>Ingle, D. J.</dc:creator>
<dc:date>2022-10-28</dc:date>
<dc:identifier>doi:10.1101/2022.10.27.514108</dc:identifier>
<dc:title><![CDATA[Tracking the horizontal transfer of plasmids in Shigella sonnei and Shigella flexneri using phylogenetics]]></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/10.1101/2022.11.02.514777v1?rss=1">
<title>
<![CDATA[
An information theoretic approach to detecting spatially varying genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.02.514777v1?rss=1</link>
<description><![CDATA[
Identifying genes with spatially coherent expression patterns is a key task in spatial transcriptomics. We adopt an information theoretic perspective on this problem by equating the degree of spatial coherence with the mutual information between nearby expression measurements. To avoid the notoriously difficult problem of computing mutual information, we use modern methods of approximation, in a method we call maximization of spatial information (Maxspin). As well as being highly scalable, we demonstrate improved accuracy across several spatial transcriptomics platforms and a variety of simulations when compared to both existing specialized methods and traditional spatial statistics methods. We use the method to analyze a renal cell carcinoma sample profiled using CosMx Spatial Molecular Imaging, revealing previously undescribed gene expression patterns.
]]></description>
<dc:creator>Jones, D. C.</dc:creator>
<dc:creator>Danaher, P.</dc:creator>
<dc:creator>Kim, Y.</dc:creator>
<dc:creator>Beechem, J. M.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:date>2022-11-03</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514777</dc:identifier>
<dc:title><![CDATA[An information theoretic approach to detecting spatially varying genes]]></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/10.1101/2022.11.03.515041v1?rss=1">
<title>
<![CDATA[
A Virus-Packageable CRISPR System Identifies Host Dependency Factors Across Multiple HIV-1 Strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.03.515041v1?rss=1</link>
<description><![CDATA[
At each stage of the HIV life cycle, host cellular proteins are hijacked by the virus to establish and enhance infection. We adapted the virus packageable HIV-CRISPR screening technology at a genome-wide scale to comprehensively identify host factors that affect HIV replication in a human T cell line. Using a smaller, targeted HIV Dependency Factor (HIVDEP) sub-library, we then performed screens across multiple HIV strains representing different clades and with different biological properties to define which T cell host factors are strain-specific versus which ones are important across all HIV strains and different clades. Nearly 90% genes selected across multiple host pathways validated in subsequent assays as bona fide host dependency factors including numerous proteins not previously reported to play role in HIV biology such as UBE2M, MBNL1, FBXW7, PELP1, SLC39A7, and others. Our ranked list of screen hits across multiple viral strains form a resource of HIV dependency factors for future investigation of host proteins involved in HIV biology.
]]></description>
<dc:creator>Montoya, V. R.</dc:creator>
<dc:creator>Ready, T. M.</dc:creator>
<dc:creator>Felton, A.</dc:creator>
<dc:creator>Fine, S. R.</dc:creator>
<dc:creator>OhAinle, M.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.03.515041</dc:identifier>
<dc:title><![CDATA[A Virus-Packageable CRISPR System Identifies Host Dependency Factors Across Multiple HIV-1 Strains]]></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/10.1101/2022.11.03.515066v1?rss=1">
<title>
<![CDATA[
Broad compatibility between yeast UAS elements and core promoters and identification of promoter elements that determine cofactor specificity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.03.515066v1?rss=1</link>
<description><![CDATA[
Three general classes of yeast protein-coding genes are distinguished by their dependence on the transcription cofactors TFIID, SAGA and Mediator (MED) Tail, but little is known about whether this dependence is determined by the core promoter, Upstream activation sites (UASs), or other gene features. It is also unclear whether UASs can broadly activate transcription from the different promoter classes or whether efficient transcription requires matching UASs and promoters of similar gene class. Here we measure transcription and cofactor specificity for tens of thousands of UAS-core promoter combinations. We find that <5% of UASs display strong core promoter specificity while most UASs can broadly activate promoters regardless of regulatory class. However, we find that matching UASs and promoters from the same gene class is generally important for optimal expression. From examining the cofactor dependence of this large UAS-promoter set, we find that sensitivity to rapid depletion of MED Tail or SAGA is dependent on the identity of both UAS and promoter while dependence on TFIID localizes to only the core promoter. Our results explain why transcription factor-mediated MED recruitment to the UAS does not always result in Tail-dependent transcription and highlight the role of TATA and TATA-like promoter sequences in MED Tail function.
]]></description>
<dc:creator>Schofield, J. A.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.03.515066</dc:identifier>
<dc:title><![CDATA[Broad compatibility between yeast UAS elements and core promoters and identification of promoter elements that determine cofactor specificity]]></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/10.1101/2022.11.05.515252v1?rss=1">
<title>
<![CDATA[
G Protein-Coupled Receptor Kinase-2 (GRK-2) regulates exploration through neuropeptide signaling in Caenorhabditis elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.05.515252v1?rss=1</link>
<description><![CDATA[
Animals alter their behavior in manners that depend on environmental conditions as well as their developmental and metabolic states. For example, C. elegans is quiescent during larval molts or during conditions of satiety. By contrast, worms enter an exploration state when removed from food. Sensory perception influences movement quiescence (defined as a lack of body movement), as well as the expression of additional locomotor states in C. elegans that are associated with increased or reduced locomotion activity, such as roaming (exploration behavior) and dwelling (local search). Here we find that movement quiescence is enhanced, and exploration behavior is reduced in G protein-coupled receptor kinase grk-2 mutant animals. grk-2 was previously shown to act in chemosensation, locomotion, and egg-laying behaviors. Using neuron-specific rescuing experiments, we show that GRK-2 acts in multiple ciliated chemosensory neurons to control exploration behavior. grk-2 acts in opposite ways from the cGMP-dependent protein kinase gene egl-4 to control movement quiescence and exploration behavior. Analysis of mutants with defects in ciliated sensory neurons indicates that grk-2 and the cilium-structure mutants act in the same pathway to control exploration behavior. We find that GRK-2 controls exploration behavior in an opposite manner from the neuropeptide receptor NPR-1 and the neuropeptides FLP-1 and FLP-18. Finally, we show that secretion of the FLP-1 neuropeptide is negatively regulated by GRK-2 and that overexpression of FLP-1 reduces exploration behavior. These results define neurons and molecular pathways that modulate movement quiescence and exploration behavior.

Author summaryMany modulatory neurotransmitters affect behavior by binding to G protein-coupled receptors (GPCRs) and initiating signals that modify neuronal activity. GPCRs are regulated by G protein-coupled receptor kinases (GRKs). GRKs phosphorylate and promote the inactivation of GPCRs. Here we identify GRK-2 as a regulator of distinct locomotor states in C. elegans. We find that GRK-2 acts in olfactory sensory neurons to promote exploration and suppress movement quiescence. Additionally, we show that GRK-2 acts in opposition to a neuropeptide signaling pathway that acts in interneurons. Thus, this study demonstrates critical roles for GRK-2 in regulating neuromodulatory signaling and locomotor behavior.
]]></description>
<dc:creator>Davis, K.</dc:creator>
<dc:creator>Mitchell, C.</dc:creator>
<dc:creator>Podhaisky, O.</dc:creator>
<dc:creator>Bai, J.</dc:creator>
<dc:creator>Raizen, D. M.</dc:creator>
<dc:creator>Ailion, M.</dc:creator>
<dc:creator>Topalidou, I.</dc:creator>
<dc:date>2022-11-05</dc:date>
<dc:identifier>doi:10.1101/2022.11.05.515252</dc:identifier>
<dc:title><![CDATA[G Protein-Coupled Receptor Kinase-2 (GRK-2) regulates exploration through neuropeptide signaling in Caenorhabditis elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-05</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/10.1101/2022.11.16.516846v1?rss=1">
<title>
<![CDATA[
The Centralspindlin proteins Pavarotti and Tumbleweed work with WASH to regulate Nuclear Envelope budding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.16.516846v1?rss=1</link>
<description><![CDATA[
Nuclear envelope (NE) budding is a nuclear pore independent nuclear export pathway, analogous to the egress of herpesviruses, and required for protein quality control, synapse development and mitochondrial integrity. The physical formation of NE buds is dependent on the Wiskott-Aldrich Syndrome protein Wash, its regulatory complex (SHRC), and Arp2/3, and requires Washs actin nucleation activity. However, the machinery governing cargo recruitment and organization within the NE bud remains unknown. Here, we identify Pavarotti (Pav) and Tumbleweed (Tum) as new molecular components of NE budding. Pav and Tum interact directly with Wash and define a second nuclear Wash-containing complex required for NE budding. Interestingly, we find that the actin bundling activities of Wash and Pav are required, suggesting a structural role in the physical and/or organizational aspects of NE buds. Thus, Pav and Tum are providing exciting new entry points into the physical machineries of this alternative nuclear export pathway for large cargos during cell differentiation and development.
]]></description>
<dc:creator>Davidson, K. A.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Verboon, J. M.</dc:creator>
<dc:creator>Parkhurst, S. M.</dc:creator>
<dc:date>2022-11-17</dc:date>
<dc:identifier>doi:10.1101/2022.11.16.516846</dc:identifier>
<dc:title><![CDATA[The Centralspindlin proteins Pavarotti and Tumbleweed work with WASH to regulate Nuclear Envelope budding]]></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/10.1101/2022.11.29.518420v1?rss=1">
<title>
<![CDATA[
FBXO42 activity is required to prevent mitotic arrest, spindle assembly checkpoint activation, and lethality in glioblastoma and other cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.29.518420v1?rss=1</link>
<description><![CDATA[
Glioblastoma (GBM) is the most common and aggressive brain tumor in adults. To identify genes differentially required for the viability of GBM stem-like cells (GSCs), we performed functional genomic lethality screens comparing GSCs and control human neural stem cells. Among top scoring hits in a subset of GBM cells was the F-box-containing gene FBXO42, which was also essential in [~]15% of cell lines derived from a broad range of cancers. Mechanistic studies revealed that, in sensitive cells, FBXO42 activity prevents chromosome alignment defects, mitotic cell cycle arrest, and cell death. The cell cycle arrest, but not the cell death, triggered by FBXO42 inactivation could be suppressed by brief exposure to a chemical inhibitor of Mps1, a key spindle assembly checkpoint (SAC) kinase. FBXO42s cancer-essential function requires its F-box and Kelch domains, which are necessary for FBXO42s substrate recognition and targeting by SCF ubiquitin ligase complex. However, none of FBXO42s previously proposed targets, including ING4, p53, and RBPJ, were responsible for the observed phenotypes. Instead, our results suggest that FBOX42 activity suppresses the accumulation of one or more proteins that perturb chromosome-microtubule dynamics in cancer cells, which, in turn, leads to induction of the SAC and cell death.
]]></description>
<dc:creator>Hoellerbauer, P.</dc:creator>
<dc:creator>Kufeld, M.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Girard, E.</dc:creator>
<dc:creator>Herman, J.</dc:creator>
<dc:creator>Olson, J.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:date>2022-12-01</dc:date>
<dc:identifier>doi:10.1101/2022.11.29.518420</dc:identifier>
<dc:title><![CDATA[FBXO42 activity is required to prevent mitotic arrest, spindle assembly checkpoint activation, and lethality in glioblastoma and other cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.06.519368v1?rss=1">
<title>
<![CDATA[
Novel populations of Tr1 cells contribute to the resolution of acute influenza A virus infection. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.06.519368v1?rss=1</link>
<description><![CDATA[
Type I regulatory (Tr1) cells contribute to immune suppression in the context of chronic infection, autoimmunity, and transplant tolerance. However, their physiological relevance in the resolution of acute respiratory infection is not understood. Here, we identify Tr1 cells accumulating in the lung parenchyma during resolution of the response to sublethal influenza A virus infection in mice. Tr1 cells were dependent on IL-27R and in their absence recovery from IAV-induced weight loss is impaired. Notably, these Tr1 cells did not necessarily co-express the typical Tr1 markers LAG-3 and CD49b, with four distinct populations of Tr1 cells apparent in the lungs. Each population was suppressive and were differentially dependent on IL-10 to mediate suppression. Transcriptional analysis revealed a core Tr1 gene signature in each population and distinct expression profiles indicative of different states of activation and differentiation. Finally, sort-transfer experiments indicated non-linear plasticity between these subsets of Tr1 cells. Together, these data support Tr1 cells contributing to the resolution of acute inflammation and define novel Tr1 cell phenotypes in acute infection.
]]></description>
<dc:creator>Abbott, C. A.</dc:creator>
<dc:creator>Freimayer, E. L.</dc:creator>
<dc:creator>Tyllis, T. S.</dc:creator>
<dc:creator>Norton, T. S.</dc:creator>
<dc:creator>Alsharifi, M.</dc:creator>
<dc:creator>Heng, A. H.</dc:creator>
<dc:creator>Pederson, S. M.</dc:creator>
<dc:creator>Qu, Z.</dc:creator>
<dc:creator>Armstrong, M.</dc:creator>
<dc:creator>Hill, G. R.</dc:creator>
<dc:creator>McColl, S. R.</dc:creator>
<dc:creator>Comerford, I.</dc:creator>
<dc:date>2022-12-06</dc:date>
<dc:identifier>doi:10.1101/2022.12.06.519368</dc:identifier>
<dc:title><![CDATA[Novel populations of Tr1 cells contribute to the resolution of acute influenza A virus infection.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.08.519635v1?rss=1">
<title>
<![CDATA[
Statistical inference reveals the role of length, breathing, and nucleotide identity in V(D)J nucleotide trimming 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.08.519635v1?rss=1</link>
<description><![CDATA[
To appropriately defend against a wide array of pathogens, humans somatically generate highly diverse repertoires of B cell and T cell receptors (BCRs and TCRs) through a random process called V(D)J recombination. Receptor diversity is achieved during this process through both the combinatorial assembly of V(D)J-genes and the junctional deletion and insertion of nucleotides. While the Artemis protein is often regarded as the main nuclease involved in V(D)J recombination, the exact mechanism of nucleotide trimming is not understood. Using a previously-published TCR{beta} repertoire sequencing data set, we have designed a flexible probabilistic model of nucleotide trimming that allows us to explore various mechanistically-interpretable sequence-level features. We show that local sequence context and the capacity for sequence-breathing, together, can most accurately predict the trimming probabilities of a given V-gene sequence. This model suggests that double-stranded DNA needs to be able to "breathe" for trimming to occur, and provides evidence of a sequence motif that appears to get preferentially trimmed, independent of breathing. Further, we find that the inferred mechanistic coefficients from this model provide accurate prediction for V- and J-gene sequences from other adaptive immune receptor loci. These results refine our understanding of how the Artemis nuclease may function to trim nucleotides during V(D)J recombination and provide another step towards understanding how V(D)J recombination generates diverse receptors and supports a powerful, unique immune response in healthy humans.
]]></description>
<dc:creator>Russell, M. L.</dc:creator>
<dc:creator>Simon, N.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2022-12-12</dc:date>
<dc:identifier>doi:10.1101/2022.12.08.519635</dc:identifier>
<dc:title><![CDATA[Statistical inference reveals the role of length, breathing, and nucleotide identity in V(D)J nucleotide trimming]]></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/10.1101/2022.12.15.520606v1?rss=1">
<title>
<![CDATA[
Identification of broad, potent antibodies to functionally constrained regions of SARS-CoV-2 spike following a breakthrough infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.15.520606v1?rss=1</link>
<description><![CDATA[
The antiviral benefit of antibodies can be compromised by viral escape especially for rapidly evolving viruses. Therefore, durable, effective antibodies must be both broad and potent to counter newly emerging, diverse strains. Discovery of such antibodies is critically important for SARS-CoV-2 as the global emergence of new variants of concern (VOC) has compromised the efficacy of therapeutic antibodies and vaccines. We describe a collection of broad and potent neutralizing monoclonal antibodies (mAbs) isolated from an individual who experienced a breakthrough infection with the Delta VOC. Four mAbs potently neutralize the Wuhan-Hu-1 vaccine strain, the Delta VOC, and also retain potency against the Omicron VOCs through BA.4/BA.5 in both pseudovirus-based and authentic virus assays. Three mAbs also retain potency to recently circulating VOCs XBB.1.5 and BQ.1.1 and one also potently neutralizes SARS-CoV-1. The potency of these mAbs was greater against Omicron VOCs than all but one of the mAbs that had been approved for therapeutic applications. The mAbs target distinct epitopes on the spike glycoprotein, three in the receptor binding domain (RBD) and one in an invariant region downstream of the RBD in subdomain 1 (SD1). The escape pathways we defined at single amino acid resolution with deep mutational scanning show they target conserved, functionally constrained regions of the glycoprotein, suggesting escape could incur a fitness cost. Overall, these mAbs are novel in their breadth across VOCs, their epitope specificity, and include a highly potent mAb targeting a rare epitope outside of the RBD in SD1.

Significance StatementSARS-CoV-2 infections can result in diverse clinical outcomes, including severe disease. Monoclonal antibodies (mAbs) have been used therapeutically to treat infection, but the emergence of variants has compromised their efficacy. Thus, identifying mAbs that are more durable in the face of SARS-CoV-2 evolution is a pressing need. Here, we describe four new mAbs isolated from a Delta-breakthrough infection, that can potently neutralize diverse variants, including multiple Omicron variants. In addition, one mAb shows broader activity against coronaviruses. The breadth of these mAbs is due to their focus on highly conserved regions of the viral protein antigen, including regions that are required for the virus to enter the cell. These properties make them promising candidates for therapeutic use.
]]></description>
<dc:creator>Guenthoer, J.</dc:creator>
<dc:creator>Lilly, M.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Lovendahl, K. N.</dc:creator>
<dc:creator>Croft, J. T.</dc:creator>
<dc:creator>Stoddard, C. I.</dc:creator>
<dc:creator>Chohan, V.</dc:creator>
<dc:creator>Ding, S.</dc:creator>
<dc:creator>Ruiz, F.</dc:creator>
<dc:creator>Kopp, M. S.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Lee, K. K.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2022-12-16</dc:date>
<dc:identifier>doi:10.1101/2022.12.15.520606</dc:identifier>
<dc:title><![CDATA[Identification of broad, potent antibodies to functionally constrained regions of SARS-CoV-2 spike following a breakthrough infection]]></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/10.1101/2022.12.19.521109v1?rss=1">
<title>
<![CDATA[
Focal adhesion assembly requires phosphorylation signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.19.521109v1?rss=1</link>
<description><![CDATA[
Integrin-mediated cell attachment rapidly induces tyrosine kinase signaling. Despite years of research, the role of this signaling in integrin activation and focal adhesion assembly is unclear. We provide evidence that the Src-family kinase (SFK) substrate Cas (Crk-associated substrate, p130Cas, BCAR1) is phosphorylated, and associated with its effectors, Crk/CrkL, in clusters that are precursors of focal adhesions. The initial phospho-Cas clusters contain integrin {beta}1 in its inactive, bent closed, conformation. Later, phospho-Cas and total Cas levels decrease as integrin {beta}1 is activated and core focal adhesion proteins including vinculin, talin, kindlin and paxillin are recruited. Cas is required for cell spreading and focal adhesion assembly in epithelial and fibroblast cells on collagen and fibronectin. Cas cluster formation requires Cas, Crk/CrkL, SFKs and Rac1 but not vinculin. Rac1 provides positive feedback onto Cas through reactive oxygen, opposed by negative feedback from the ubiquitin proteasome system. The results suggest a two-step model for focal adhesion assembly in which clusters of phospho-Cas, effectors and inactive integrin {beta}1 grow through positive feedback prior to integrin activation and recruitment of core focal adhesion proteins.
]]></description>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Stainer, A.</dc:creator>
<dc:creator>Dubrulle, J.</dc:creator>
<dc:creator>Simpkins, C.</dc:creator>
<dc:creator>Cooper, J.</dc:creator>
<dc:date>2022-12-19</dc:date>
<dc:identifier>doi:10.1101/2022.12.19.521109</dc:identifier>
<dc:title><![CDATA[Focal adhesion assembly requires phosphorylation signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.20.521287v1?rss=1">
<title>
<![CDATA[
Single-cell profiling uncovers a Muc4-expressing metaplastic gastric cell type sustained by Helicobacter pylori-specific inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.20.521287v1?rss=1</link>
<description><![CDATA[
AbstractMechanisms for Helicobacter pylori (Hp)-driven stomach cancer are not fully understood. In a transgenic mouse model of gastric preneoplasia, concomitant Hp infection and induction of constitutively active KRAS (Hp+KRAS+) alters metaplasia phenotypes and elicits greater inflammation than either perturbation alone. Gastric single-cell RNA-seq showed that Hp+KRAS+ mice had a large population of metaplastic pit cells that expressed the intestinal mucin Muc4 and the growth factor amphiregulin. Metaplastic pit cells were associated with macrophage and T cell inflammation and prevented by gastric immunosuppression. Lineage tracing showed that Muc4 was not dependent on cell-intrinsic KRAS activity, and lineage-derived cells had a limited propensity for growth as organoids, demonstrating that metaplastic pit cells are largely not self- renewing. Finally, MUC4 expression was significantly associated with proliferation in human gastric cancer samples. These studies identify an Hp-associated metaplastic pit cell lineage, also found in human gastric cancer tissues, whose expansion is driven by Hp-dependent inflammation.

Statement of SignificanceUsing a mouse model, we have delineated metaplastic pit cells as a pre-cancerous cell type whose expansion requires H. pylori-driven inflammation. In humans, metaplastic pit cells show enhanced proliferation as well as enrichment in precancer and early cancer tissues, highlighting an early step in the gastric metaplasia to cancer cascade.
]]></description>
<dc:creator>O'Brien, V. P.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Shenoy, M. K.</dc:creator>
<dc:creator>Finak, G.</dc:creator>
<dc:creator>Young, W. C.</dc:creator>
<dc:creator>Dubrulle, J.</dc:creator>
<dc:creator>Koch, L.</dc:creator>
<dc:creator>Martinez, A. E. R.</dc:creator>
<dc:creator>Snow, J. A.</dc:creator>
<dc:creator>Williams, J.</dc:creator>
<dc:creator>Donato, E.</dc:creator>
<dc:creator>Batra, S. K.</dc:creator>
<dc:creator>Yeung, C. C.</dc:creator>
<dc:creator>Bullman, S.</dc:creator>
<dc:creator>Koch, M. A.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Salama, N. R.</dc:creator>
<dc:date>2022-12-20</dc:date>
<dc:identifier>doi:10.1101/2022.12.20.521287</dc:identifier>
<dc:title><![CDATA[Single-cell profiling uncovers a Muc4-expressing metaplastic gastric cell type sustained by Helicobacter pylori-specific inflammation]]></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/10.1101/2022.12.22.521701v1?rss=1">
<title>
<![CDATA[
Recursive splicing discovery using lariats in total RNA sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.22.521701v1?rss=1</link>
<description><![CDATA[
Recursive splicing is a noncanonical splicing mechanism in which an intron is removed in segments via multiple splicing reactions. Relatively few recursive splice sites have been identified with high confidence in human introns, and more comprehensive analyses are needed to better characterize where recursive splicing happens and whether or not it has a regulatory function. In this study, we use an unbiased approach utilizing intron lariats to search for recursive splice sites in constitutive introns and alternative exons in the human transcriptome. We find evidence for recursive splicing in a broader range of intron sizes than previously reported and detail a new location for recursive splicing at the distal ends of cassette exons. Additionally, we identify evidence for the conservation of these recursive splice sites among higher vertebrates and the use of these sites to influence alternative exon exclusion. Together, our data demonstrate the prevalence of recursive splicing and its potential influence on gene expression through alternatively spliced isoforms.
]]></description>
<dc:creator>Hoppe, E. R.</dc:creator>
<dc:creator>Udy, D. B.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:date>2022-12-23</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521701</dc:identifier>
<dc:title><![CDATA[Recursive splicing discovery using lariats in total RNA sequencing]]></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/10.1101/2022.12.26.521704v1?rss=1">
<title>
<![CDATA[
Epithelia delimits glial apical polarity against mechanical shear to maintain glia-neuron architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.26.521704v1?rss=1</link>
<description><![CDATA[
For an organ to maintain proper architecture and function, its different component cell-types must coordinate their cell-shapes with each other through life. While cell-intrinsic developmental mechanisms driving homotypic cell-cell coordination are known, how heterotypic cells collectively regulate cell-shape is less-clear. We report that, in a sense-organ, epithelial cells delimit and maintain polarity domains of contacting glia, and thereby, associated neuron shapes throughout life. Briefly, Hsp co-chaperone UNC-23/BAG2 keeps epithelial apical domains from deforming with animal movement. Epithelial apical domains stretch aberrantly and progressively in adult unc-23 mutant animals, which in an FGFR-dependent manner, dislocates glial apical cytoskeleton proteins SMA-1/{beta}H-Spectrin and actin. This alters glial apical polarity and cell shape, and concomitantly, associated neuron-ending shape. Notably, UNC-23 acts temporally at a developmental critical period to maintain glia-neuron shape in adults, and spatially within a defined anatomical zone. Lastly, intervention in either epithelia, glia or neuron ameliorate or phenocopy unc-23 neural defects. Epi/endothelia resist mechanical stress and contact glia-neuron units across central/peripheral nervous systems and species, and all components of the identified molecular pathway are conserved and disease-relevant. Thus, we posit that analogous epithelia-glia mechanobiological coupling may broadly regulate glia-neuron shapes through animal life.
]]></description>
<dc:creator>Martin, C. G.</dc:creator>
<dc:creator>Bent, J. S.</dc:creator>
<dc:creator>Singhvi, A.</dc:creator>
<dc:date>2022-12-27</dc:date>
<dc:identifier>doi:10.1101/2022.12.26.521704</dc:identifier>
<dc:title><![CDATA[Epithelia delimits glial apical polarity against mechanical shear to maintain glia-neuron architecture]]></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/10.1101/2023.01.03.522658v1?rss=1">
<title>
<![CDATA[
An RNA seq-based reference landscape of human normal and neoplastic brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.03.522658v1?rss=1</link>
<description><![CDATA[
In order to better understand the relationship between normal and neoplastic brain, we combined five publicly available large-scale datasets, correcting for batch effects and applying Uniform Manifold Approximation and Projection (UMAP) to RNA-seq data. We assembled a reference Brain-UMAP including 702 adult gliomas, 802 pediatric tumors and 1409 healthy normal brain samples, which can be utilized to investigate the wealth of information obtained from combining several publicly available datasets to study a single organ site. Normal brain regions and tumor types create distinct clusters and because the landscape is generated by RNA seq, comparative gene expression profiles and gene ontology patterns are readily evident. To our knowledge, this is the first meta-analysis that allows for comparison of gene expression and pathways of interest across adult gliomas, pediatric brain tumors, and normal brain regions. We provide access to this resource via the open source, interactive online tool Oncoscape, where the scientific community can readily visualize clinical metadata, gene expression patterns, gene fusions, mutations, and copy number patterns for individual genes and pathway over this reference landscape.
]]></description>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:creator>Jensen, M.</dc:creator>
<dc:creator>Nuechterlein, N.</dc:creator>
<dc:creator>Pattwell, S. S.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:date>2023-01-03</dc:date>
<dc:identifier>doi:10.1101/2023.01.03.522658</dc:identifier>
<dc:title><![CDATA[An RNA seq-based reference landscape of human normal and neoplastic brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.05.522885v1?rss=1">
<title>
<![CDATA[
Functional genomic analysis of adult and pediatric brain tumor isolates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.05.522885v1?rss=1</link>
<description><![CDATA[
BackgroundAdult and pediatric tumors display stark differences in their mutation spectra and chromosome alterations. Here, we attempted to identify common and unique gene dependencies and their associated biomarkers among adult and pediatric tumor isolates using functional genetic lethal screens and computational modeling.

MethodsWe performed CRISRP-Cas9 lethality screens in two adult glioblastoma (GBM) tumor isolates and five pediatric brain tumor isolates representing atypical teratoid rhabdoid tumors (ATRT), diffuse intrinsic pontine glioma, GBM, and medulloblastoma. We then integrated the screen results with machine learning-based gene-dependency models generated from data from >900 cancer cell lines.

ResultsWe found that >50% of candidate dependencies of 280 identified were shared between adult GBM tumors and individual pediatric tumor isolates. 68% of screen hits were found as nodes in our network models, along with shared and tumor-specific predictors of gene dependencies. We investigated network predictors associated with ADAR, EFR3A, FGFR1 (pediatric-specific), and SMARCC2 (ATRT-specific) gene dependency among our tumor isolates.

ConclusionsThe results suggest that, despite harboring disparate genomic signatures, adult and pediatric tumor isolates share a preponderance of genetic dependences. Further, combining data from primary brain tumor lethality screens with large cancer cell line datasets produced valuable insights into biomarkers of gene dependency, even for rare cancers.

Importance of the StudyOur results demonstrate that large cancer cell lines data sets can be computationally mined to identify known and novel gene dependency relationships in adult and pediatric human brain tumor isolates. Gene dependency networks and lethality screen results represent a key resource for neuro-oncology and cancer research communities. We also highlight some of the challenges and limitations of this approach.
]]></description>
<dc:creator>Hoellerbauer, P.</dc:creator>
<dc:creator>Biery, M.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Rao, Y.</dc:creator>
<dc:creator>Girard, E.</dc:creator>
<dc:creator>Mitchell, K.</dc:creator>
<dc:creator>Dighe, P.</dc:creator>
<dc:creator>Kufeld, M.</dc:creator>
<dc:creator>Kuppers, D.</dc:creator>
<dc:creator>Herman, J.</dc:creator>
<dc:creator>Holland, E.</dc:creator>
<dc:creator>Soroceanu, L.</dc:creator>
<dc:creator>Vitanza, N.</dc:creator>
<dc:creator>Olson, J.</dc:creator>
<dc:creator>Pritchard, J.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:date>2023-01-06</dc:date>
<dc:identifier>doi:10.1101/2023.01.05.522885</dc:identifier>
<dc:title><![CDATA[Functional genomic analysis of adult and pediatric brain tumor isolates]]></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/10.1101/2023.01.05.522947v1?rss=1">
<title>
<![CDATA[
Somatic mutation but not aneuploidy differentiates lung cancer in never-smokers and smokers. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.05.522947v1?rss=1</link>
<description><![CDATA[
Lung cancer in never-smokers disproportionately affects older women. To understand the mutational landscape of this cohort, we performed detailed genome characterization of 73 lung adenocarcinomas from participants of the Womens Health Initiative (WHI). We find enrichment of EGFR mutations in never-/light-smokers and KRAS mutations in heavy smokers as expected, but we also show that the specific variants of these genes differ by smoking status, with important therapeutic implications. Mutational signature analysis revealed signatures of clock, APOBEC, and DNA repair deficiency in never-/light-smokers; however, the mutational load of these signatures did not differ significantly from those found in smokers. Last, tumors from both smokers and never-/light-smokers shared copy number subtypes, with no significant differences in aneuploidy. Thus, the genomic landscape of lung cancer in never-/light-smokers and smokers is predominantly differentiated by somatic mutations and not copy number alterations.
]]></description>
<dc:creator>Moorthi, S.</dc:creator>
<dc:creator>Paguirigan, A.</dc:creator>
<dc:creator>Ko, M.</dc:creator>
<dc:creator>Pettinger, M.</dc:creator>
<dc:creator>Hoge, A. C. H.</dc:creator>
<dc:creator>Nag, A.</dc:creator>
<dc:creator>Patel, N. A.</dc:creator>
<dc:creator>Wu, F.</dc:creator>
<dc:creator>Sather, C.</dc:creator>
<dc:creator>Fitzgibbon, M. P.</dc:creator>
<dc:creator>Thorner, A. R.</dc:creator>
<dc:creator>Anderson, G. L.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:creator>Berger, A. H.</dc:creator>
<dc:date>2023-01-06</dc:date>
<dc:identifier>doi:10.1101/2023.01.05.522947</dc:identifier>
<dc:title><![CDATA[Somatic mutation but not aneuploidy differentiates lung cancer in never-smokers and smokers.]]></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/10.1101/2023.01.07.522490v1?rss=1">
<title>
<![CDATA[
Genetics of Latin American Diversity (GLAD) Project: insights into population genetics and association studies in recently admixed groups in the Americas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.07.522490v1?rss=1</link>
<description><![CDATA[
Latin America is underrepresented in genetic studies, which can exacerbate disparities in personalized genomic medicine. However, genetic data of thousands of Latin Americans are already publicly available, but require a bureaucratic maze to navigate all the data access and consenting issues. We present the Genetics of Latin American Diversity (GLAD) Project, a platform that compiles genome-wide information of 54,077 Latin Americans from 39 studies representing 45 geographical regions. Through GLAD, we identified heterogeneous ancestry composition and recent gene-flow across the Americas. Also, we developed a simulated-annealing-based algorithm to match the genetic background of external samples to our database and share summary statistics without transferring individual-level data. Finally, we demonstrate the potential of GLAD as a critical resource for evaluating statistical genetic softwares in the presence of admixture. By making this resource available, we promote genomic research in Latin Americans and contribute to the promises of personalized medicine to more people.
]]></description>
<dc:creator>Borda, V.</dc:creator>
<dc:creator>Loesch, D. P.</dc:creator>
<dc:creator>Guo, B.</dc:creator>
<dc:creator>Laboulaye, R.</dc:creator>
<dc:creator>Veliz-Otani, D.</dc:creator>
<dc:creator>French-Kwawu, J. N.</dc:creator>
<dc:creator>Peixoto Leal, T.</dc:creator>
<dc:creator>Gogarten, S. M.</dc:creator>
<dc:creator>Ikpe, S.</dc:creator>
<dc:creator>Gouveia, M. H.</dc:creator>
<dc:creator>Mendes, M.</dc:creator>
<dc:creator>Abecasis, G. R.</dc:creator>
<dc:creator>Alvim, I.</dc:creator>
<dc:creator>Arboleda-Bustos, C. E.</dc:creator>
<dc:creator>Arboleda, G.</dc:creator>
<dc:creator>Arboleda, H.</dc:creator>
<dc:creator>Barreto, M. L.</dc:creator>
<dc:creator>Barwick, L.</dc:creator>
<dc:creator>Bezzera, M. A.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Borges, V.</dc:creator>
<dc:creator>Caceres, O.</dc:creator>
<dc:creator>Cai, J.</dc:creator>
<dc:creator>Chana-Cuevas, P.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Custer, B.</dc:creator>
<dc:creator>Dean, M.</dc:creator>
<dc:creator>Dinardo, C.</dc:creator>
<dc:creator>Domingos, I.</dc:creator>
<dc:creator>Duggirala, R.</dc:creator>
<dc:creator>Dieguez, E.</dc:creator>
<dc:creator>Fernandez, W.</dc:creator>
<dc:creator>Ferraz, H. B.</dc:creator>
<dc:creator>Gilliland, F. D.</dc:creator>
<dc:creator>Guio, H.</dc:creator>
<dc:creator>Horta, B.</dc:creator>
<dc:creator>Curran, J. E.</dc:creator>
<dc:creator>Johnsen, J. M.</dc:creator>
<dc:creator>Kaplan, R. C.</dc:creator>
<dc:creator>Kelly, S.</dc:creator>
<dc:creator>Kenny, E. E.</dc:creator>
<dc:creator>Kittner, S.</dc:creator>
<dc:creator>Konkle,</dc:creator>
<dc:date>2023-01-09</dc:date>
<dc:identifier>doi:10.1101/2023.01.07.522490</dc:identifier>
<dc:title><![CDATA[Genetics of Latin American Diversity (GLAD) Project: insights into population genetics and association studies in recently admixed groups in the Americas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.09.523323v1?rss=1">
<title>
<![CDATA[
Aclarubicin stimulates RNA polymerase II elongation at closely spaced divergent promoters 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.09.523323v1?rss=1</link>
<description><![CDATA[
Anthracyclines are a class of widely prescribed anti-cancer drugs that disrupt chromatin by intercalating into DNA and enhancing nucleosome turnover. To understand the molecular consequences of anthracycline-mediated chromatin disruption, we utilized CUT&Tag to profile RNA polymerase II during anthracycline treatment in Drosophila cells. We observed that treatment with the anthracycline aclarubicin leads to elevated levels of elongating RNA polymerase II and changes in chromatin accessibility. We found that promoter proximity and orientation impacts chromatin changes during aclarubicin treatment, as closely spaced divergent promoter pairs show greater chromatin changes when compared to codirectionally-oriented tandem promoters. We also found that aclarubicin treatment changes the distribution of non-canonical DNA G-quadruplex structures both at promoters and at G-rich pericentromeric repeats. Our work suggests that the anti-cancer activity of aclarubicin is driven by the effects of nucleosome disruption on RNA polymerase II, chromatin accessibility and DNA structures.
]]></description>
<dc:creator>Wooten, M.</dc:creator>
<dc:creator>Takushi, B.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2023-01-10</dc:date>
<dc:identifier>doi:10.1101/2023.01.09.523323</dc:identifier>
<dc:title><![CDATA[Aclarubicin stimulates RNA polymerase II elongation at closely spaced divergent promoters]]></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/10.1101/2023.01.10.523535v1?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/10.1101/2023.01.19.524800v1?rss=1">
<title>
<![CDATA[
Damage-induced pyroptosis drives endogenous thymic regeneration viainduction of Foxn1 by purinergic receptor activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.19.524800v1?rss=1</link>
<description><![CDATA[
Endogenous thymic regeneration is a crucial process that allows for the renewal of immune competence following stress, infection or cytoreductive conditioning. Fully understanding the molecular mechanisms driving regeneration will uncover therapeutic targets to enhance regeneration. We previously demonstrated that high levels of homeostatic apoptosis suppress regeneration and that a reduction in the presence of damage-induced apoptotic thymocytes facilitates regeneration. Here we identified that cell-specific metabolic remodeling after ionizing radiation steers thymocytes towards mitochondrial-driven pyroptotic cell death. We further identified that a key damage-associated molecular pattern (DAMP), ATP, stimulates the cell surface purinergic receptor P2Y2 on cortical thymic epithelial cells (cTECs) acutely after damage, enhancing expression of Foxn1, the critical thymic transcription factor. Targeting the P2Y2 receptor with the agonist UTP{gamma}S promotes rapid regeneration of the thymus in vivo following acute damage. Together these data demonstrate that intrinsic metabolic regulation of pyruvate processing is a critical process driving thymus repair and identifies the P2Y2 receptor as a novel molecular therapeutic target to enhance thymus regeneration.

SUMMARYO_LIThymocytes rapidly and transiently undergo pyroptosis after acute thymic damage and promote regeneration.
C_LIO_LIDamage-induced redirection of pyruvate acutely enhances mitochondrial OXPHOS in thymocytes.
C_LIO_LIElevated mitochondrial ROS promotes pyroptosis in thymocytes after acute insult by driving caspase 1 cleavage.
C_LIO_LIExtracellular ATP release promotes Foxn1 expression in cTECs via activation of P2Y2
C_LIO_LITherapeutic targeting of the P2Y2 receptor promotes thymic regeneration.
C_LI
]]></description>
<dc:creator>Kinsella, S.</dc:creator>
<dc:creator>Evandy, C. A.</dc:creator>
<dc:creator>Cooper, K.</dc:creator>
<dc:creator>Cardinale, A.</dc:creator>
<dc:creator>Iovino, L.</dc:creator>
<dc:creator>deRoos, P.</dc:creator>
<dc:creator>Hopwo, K. S.</dc:creator>
<dc:creator>Smith, C. W.</dc:creator>
<dc:creator>Granadier, D. W.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:creator>Velardi, E.</dc:creator>
<dc:creator>Dudakov, J.</dc:creator>
<dc:date>2023-01-21</dc:date>
<dc:identifier>doi:10.1101/2023.01.19.524800</dc:identifier>
<dc:title><![CDATA[Damage-induced pyroptosis drives endogenous thymic regeneration viainduction of Foxn1 by purinergic receptor activation]]></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/10.1101/2023.01.19.524825v1?rss=1">
<title>
<![CDATA[
Oligonucleotide-directed proximity-interactome mapping (O-MAP): A unified method for discovering RNA-interacting proteins, transcripts and genomic loci in situ. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.19.524825v1?rss=1</link>
<description><![CDATA[
Throughout biology, RNA molecules form complex networks of molecular interactions that are central to their function, but remain challenging to investigate. Here, we introduce Oligonucleotide-mediated proximity-interactome MAPping (O-MAP), a straightforward method for elucidating the biomolecules near an RNA of interest, within its native cellular context. O-MAP uses programmable oligonucleotide probes to deliver proximity-biotinylating enzymes to a target RNA, enabling nearby molecules to be enriched by streptavidin pulldown. O-MAP induces exceptionally precise RNA-localized in situ biotinylation, and unlike alternative methods it enables straightforward optimization of its targeting accuracy. Using the 47S pre-ribosomal RNA and long noncoding RNA Xist as models, we develop O-MAP workflows for unbiased discovery of RNA-proximal proteins, transcripts, and genomic loci. This revealed unexpected co-compartmentalization of Xist and other chromatin-regulatory RNAs and enabled systematic characterization of nucleolar-chromatin interactions across multiple cell lines. O-MAP is portable to cultured cells, organoids, and tissues, and to RNAs of various lengths, abundances, and sequence composition. And, O-MAP requires no genetic manipulation and uses exclusively off-the-shelf parts. We therefore anticipate its application to a broad array of RNA phenomena.
]]></description>
<dc:creator>Tsue, A. F.</dc:creator>
<dc:creator>Kania, E. E.</dc:creator>
<dc:creator>Lei, D. Q.</dc:creator>
<dc:creator>Fields, R.</dc:creator>
<dc:creator>McGann, C. C.</dc:creator>
<dc:creator>Hershberg, E. A.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Kihiu, M.</dc:creator>
<dc:creator>Ong, S.-E.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:creator>Kugel, S.</dc:creator>
<dc:creator>Beliveau, B.</dc:creator>
<dc:creator>Schweppe, D. K.</dc:creator>
<dc:creator>Shechner, D. M.</dc:creator>
<dc:date>2023-01-20</dc:date>
<dc:identifier>doi:10.1101/2023.01.19.524825</dc:identifier>
<dc:title><![CDATA[Oligonucleotide-directed proximity-interactome mapping (O-MAP): A unified method for discovering RNA-interacting proteins, transcripts and genomic loci in situ.]]></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/10.1101/2023.01.20.524981v1?rss=1">
<title>
<![CDATA[
Single molecule visualization of native centromeric nucleosome formation reveals coordinated deposition by kinetochore proteins and centromere DNA sequence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.20.524981v1?rss=1</link>
<description><![CDATA[
Eukaryotic chromosome segregation requires the kinetochore, a megadalton-sized machine that forms on specialized centromeric chromatin containing CENP-A, a histone H3 variant. CENP-A deposition requires a chaperone protein HJURP that targets it to the centromere, but it has remained unclear whether HJURP has additional functions beyond CENP-A targeting and why high AT DNA content, which disfavors nucleosome assembly, is widely conserved at centromeres. To overcome the difficulties of studying nucleosome formation in vivo, we developed a microscopy assay that enables direct observation of de novo centromeric nucleosome recruitment and maintenance with single molecule resolution. Using this assay, we discover that CENP-A can arrive at centromeres without its dedicated centromere-specific chaperone HJURP, but stable incorporation depends on HJURP and additional DNA-binding proteins of the inner kinetochore. We also show that homopolymer AT runs in the yeast centromeres are essential for efficient CENP-A deposition. Together, our findings reveal requirements for stable nucleosome formation and provide a foundation for further studies of the assembly and dynamics of native kinetochore complexes.
]]></description>
<dc:creator>Popchock, A.</dc:creator>
<dc:creator>Larson, J.</dc:creator>
<dc:creator>Dubrulle, J.</dc:creator>
<dc:creator>Asbury, C. L.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2023-01-21</dc:date>
<dc:identifier>doi:10.1101/2023.01.20.524981</dc:identifier>
<dc:title><![CDATA[Single molecule visualization of native centromeric nucleosome formation reveals coordinated deposition by kinetochore proteins and centromere DNA sequence]]></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/10.1101/2023.01.22.524153v1?rss=1">
<title>
<![CDATA[
Mitochondrial diabetes in mice expressing a dominant-negative allele of nuclear respiratory factor-1 (NRF-1) in pancreatic and β-cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.22.524153v1?rss=1</link>
<description><![CDATA[
Genetic polymorphisms in nuclear respiratory factor-1 (NRF1), a key transcriptional regulator of nuclear-encoded mitochondrial proteins, have been linked to diabetes. Homozygous deletion of Nrf1 is embryonic lethal in mice. Our goal was to generate mice with {beta}-cell-specific reduction in NRF1 function to investigate the relationship between NRF1 and diabetes. We report the generation of mice expressing a dominant-negative allele of Nrf1 (DNNRF1) in pancreatic {beta}-cells. Heterozygous transgenic mice had high fed blood glucose levels detected at 3 wks of age, which persisted through adulthood. Plasma insulin levels in DNNRF1 transgenic mice were reduced, while insulin sensitivity remained intact in young animals. Islet size was reduced with increased numbers of apoptotic cells, and insulin content in islets by immunohistochemistry was low. Glucose-stimulated insulin secretion in isolated islets was reduced in DNNRF1-mice, but partially rescued by KCl, suggesting that decreased mitochondrial function contributed to the insulin secretory defect. Electron micrographs demonstrated abnormal mitochondrial morphology in {beta}- cells. Expression of NRF1 target genes Tfam, T@1m and T@2m, and islet cytochrome c oxidase and succinate dehydrogenase activities were reduced in DNNRF1-mice. Rescue of mitochondrial function with low level activation of transgenic c-Myc in {beta}-cells was sufficient to restore {beta}-cell mass and prevent diabetes. This study demonstrates that reduced NRF1 function can lead to loss of {beta}-cell function and establishes a model to study the interplay between regulators of bi- genomic gene transcription in diabetes.
]]></description>
<dc:creator>Morrish, F.</dc:creator>
<dc:creator>Sweet, I. R.</dc:creator>
<dc:creator>Hockenbery, D. M.</dc:creator>
<dc:date>2023-01-22</dc:date>
<dc:identifier>doi:10.1101/2023.01.22.524153</dc:identifier>
<dc:title><![CDATA[Mitochondrial diabetes in mice expressing a dominant-negative allele of nuclear respiratory factor-1 (NRF-1) in pancreatic and β-cells]]></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/10.1101/2023.01.22.525083v1?rss=1">
<title>
<![CDATA[
RNA Polymerase II, the BAF remodeler and transcription factors synergize to evict nucleosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.22.525083v1?rss=1</link>
<description><![CDATA[
Chromatin accessibility is a hallmark of active transcription and requires ATP-dependent nucleosome remodeling by Brahma-Associated Factor (BAF). However, the mechanistic link between transcription, nucleosome remodeling, and chromatin accessibility is unclear. Here, we used a chemical-genetic approach to dissect the interplay between RNA Polymerase II (RNAPII), BAF, and DNA-sequence-specific transcription factors (TFs) in mouse embryonic stem cells. By time-resolved chromatin profiling with acute transcription block at distinct stages, we show that RNAPII promoter-proximal pausing stabilizes BAF chromatin occupancy and enhances nucleosome eviction by BAF. We find that RNAPII and BAF probe both transcriptionally active and Polycomb-repressed genomic regions and provide evidence that TFs capture transient site exposure due to nucleosome unwrapping by BAF to confer locus specificity for persistent chromatin remodeling. Our study reveals the mechanistic basis of cell-type-specific chromatin accessibility. We propose a new paradigm for how functional synergy between dynamically acting chromatin factors regulates nucleosome organization.
]]></description>
<dc:creator>Brahma, S.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2023-01-22</dc:date>
<dc:identifier>doi:10.1101/2023.01.22.525083</dc:identifier>
<dc:title><![CDATA[RNA Polymerase II, the BAF remodeler and transcription factors synergize to evict nucleosomes]]></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/10.1101/2023.02.01.526589v1?rss=1">
<title>
<![CDATA[
Polypharmacology-based kinome screen identifies new regulators of KSHV reactivation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.01.526589v1?rss=1</link>
<description><![CDATA[
Kaposis sarcoma-associated herpesvirus (KSHV) causes several human diseases including Kaposis sarcoma (KS), a leading cause of cancer in Africa and in patients with AIDS. KS tumor cells harbor KSHV predominantly in a latent form, while typically <5% contain lytic replicating virus. Because both latent and lytic stages likely contribute to cancer initiation and progression, continued dissection of host regulators of this biological switch will provide insights into fundamental pathways controlling the KSHV life cycle and related disease pathogenesis. Several cellular protein kinases have been reported to promote or restrict KSHV reactivation, but our knowledge of these signaling mediators and pathways is incomplete. We employed a polypharmacology-based kinome screen to identifiy specific kinases that regulate KSHV reactivation. Those identified by the screen and validated by knockdown experiments included several kinases that enhance lytic reactivation: ERBB2 (HER2 or neu), ERBB3 (HER3), ERBB4 (HER4), MKNK2 (MNK2), ITK, TEC, and DSTYK (RIPK5). Conversely, ERBB1 (EGFR1 or HER1), MKNK1 (MNK1) and FRK (PTK5) were found to promote the maintenance of latency. Mechanistic characterization of ERBB2 pro-lytic functions revealed a signaling connection between ERBB2 and the activation of CREB1, a transcription factor that drives KSHV lytic gene expression. These studies provided a proof-of-principle application of a polypharmacology-based kinome screen for the study of KSHV reactivation and enabled the discovery of both kinase inhibitors and specific kinases that regulate the KSHV latent-to-lytic replication switch.

Author SummaryKaposis sarcoma-associated herpesvirus (KSHV) causes Kaposis sarcoma, a cancer particularly prevalent in Africa. In cancer cells, the virus persists in a quiescent form called latency, in which only a few viral genes are made. Periodically, the virus switches into an active replicative cycle in which most of the viral genes are made and new virus is produced. What controls the switch from latency to active replication is not well understood, but cellular kinases, enzymes that control many cellular processes, have been implicated. Using a cell culture model of KSHV reactivation along with an innovative screening method that probes the effects of many cellular kinases simultaneously, we identified drugs that significantly limit KSHV reactivation, as well as specific kinases that either enhance or restrict KSHV replicative cycle. Among these were the ERBB kinases which are known to regulate growth of cancer cells. Understanding how these and other kinases contribute to the switch leading to production of more infectious virus helps us understand the mediators and mechanisms of KSHV diseases. Additionally, because kinase inhibitors are proving to be effective for treating other diseases including some cancers, identifying ones that restrict KSHV replicative cycle may lead to new approaches to treating KSHV-related diseases.
]]></description>
<dc:creator>Olson, A. T.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Ladha, A. M.</dc:creator>
<dc:creator>Lim, C. B.</dc:creator>
<dc:creator>Lagunoff, M.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:creator>Geballe, A. P.</dc:creator>
<dc:date>2023-02-01</dc:date>
<dc:identifier>doi:10.1101/2023.02.01.526589</dc:identifier>
<dc:title><![CDATA[Polypharmacology-based kinome screen identifies new regulators of KSHV reactivation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.06.527330v1?rss=1">
<title>
<![CDATA[
Elevated binding and functional antibody responses to SARS-CoV-2 in infants versus mothers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.06.527330v1?rss=1</link>
<description><![CDATA[
Infant antibody responses to viral infection can differ from those in adults. However, data on the specificity and function of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies in infants, and direct comparisons between infants and adults are limited. We characterized antibody binding and functionality in convalescent plasma from postpartum women and their infants infected with SARS-CoV-2 from a vaccine-naive prospective cohort in Nairobi, Kenya. Antibody titers against SARS-CoV-2 Spike, receptor binding domain and N-terminal domain, and Spike-expressing cell-surface staining levels were significantly higher in infants than in mothers. Plasma antibodies from mothers and infants bound to similar regions of the Spike S2 subunit, including the fusion peptide (FP) and stem helix-heptad repeat 2. However, infants displayed higher antibody levels and more consistent antibody escape pathways in the FP region compared to mothers. Finally, infants had significantly higher levels of antibody-dependent cellular cytotoxicity (ADCC), though, surprisingly, neutralization titers between infants and mothers were similar. These results suggest infants develop distinct SARS-CoV-2 binding and functional antibody repertoires and reveal age-related differences in humoral immunity to SARS-CoV-2 infection that could be relevant to protection and COVID-19 disease outcomes.
]]></description>
<dc:creator>Stoddard, C. I.</dc:creator>
<dc:creator>Sung, K.</dc:creator>
<dc:creator>Yaffe, Z. A.</dc:creator>
<dc:creator>Weight, H.</dc:creator>
<dc:creator>Beaudoin-Bussieres, G.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>Gantt, S.</dc:creator>
<dc:creator>Adhiambo, J.</dc:creator>
<dc:creator>Begnel, E. R.</dc:creator>
<dc:creator>Ojee, E.</dc:creator>
<dc:creator>Slyker, J.</dc:creator>
<dc:creator>Wamalwa, D.</dc:creator>
<dc:creator>Kinuthia, J.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Lehman, D. A.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2023-02-07</dc:date>
<dc:identifier>doi:10.1101/2023.02.06.527330</dc:identifier>
<dc:title><![CDATA[Elevated binding and functional antibody responses to SARS-CoV-2 in infants versus mothers]]></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/10.1101/2023.02.07.527545v1?rss=1">
<title>
<![CDATA[
Several cell-intrinsic effectors drive type I interferon-mediated restriction of HIV-1 in primary CD4+ T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.07.527545v1?rss=1</link>
<description><![CDATA[
Type I interferon (IFN) upregulates proteins that inhibit HIV within infected cells. Prior studies have identified IFN-stimulated genes (ISGs) that impede lab-adapted HIV in cell lines, yet the ISG(s) that mediate IFN restriction in HIV target cells, primary CD4+ T cells, are unknown. Here, we interrogate ISG restriction of primary HIV in CD4+ T cells. We performed CRISPR-knockout screens using a custom library that specifically targets ISGs expressed in CD4+ T cells and validated top hits. Our investigation identified new HIV-restricting ISGs (HM13, IGFBP2, LAP3) and found that two previously studied factors (IFI16, UBE2L6) are IFN effectors in T cells. Inactivation of these five ISGs in combination further diminished IFNs protective effect against six diverse HIV strains. This work demonstrates that IFN restriction of HIV is multifaceted, resulting from several effectors functioning collectively, and establishes a primary cell ISG screening model to identify both single and combinations of HIV-restricting ISGs.
]]></description>
<dc:creator>Itell, H. L.</dc:creator>
<dc:creator>Humes, D.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2023-02-07</dc:date>
<dc:identifier>doi:10.1101/2023.02.07.527545</dc:identifier>
<dc:title><![CDATA[Several cell-intrinsic effectors drive type I interferon-mediated restriction of HIV-1 in primary CD4+ T cells]]></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/10.1101/2023.02.18.529093v1?rss=1">
<title>
<![CDATA[
The Regulation of Nucleic Acid Vaccine Responses by the Microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.18.529093v1?rss=1</link>
<description><![CDATA[
SummaryNucleic acid vaccines, including both RNA and DNA platforms, are key technologies that have considerable promise in combating both infectious disease and cancer. However, little is known about the extrinsic factors that regulate nucleic acid vaccine responses and which may determine their effectiveness. The microbiome is recognized as a significant regulator of immune development and response, whose role in regulating some traditional vaccine platforms has recently been discovered. Using germ-free and specific-pathogen-free mouse models in combination with different protein, DNA, and mRNA vaccine regimens, we demonstrate that the microbiome is a significant regulator of nucleic acid vaccine immunogenicity. While the presence of the microbiome enhances CD8+ T cell responses to mRNA lipid nanoparticle (LNP) immunization, the microbiome suppresses immunoglobulin and CD4+ T cell responses to DNA-prime, DNA-protein-boost immunization, indicating contrasting roles for the microbiome in the regulation of these different nucleic acid vaccine platforms. In the case of mRNA-LNP vaccination, germ-free mice display reduced dendritic cell/macrophage activation that may underlie the deficient vaccine response. Our study identifies the microbiome as a relevant determinant of nucleic acid vaccine response with implications for their continued therapeutic development and deployment.
]]></description>
<dc:creator>Johnson, A. M. F.</dc:creator>
<dc:creator>Hager, K.</dc:creator>
<dc:creator>Alameh, M.-G.</dc:creator>
<dc:creator>Van, P.</dc:creator>
<dc:creator>Potchen, N.</dc:creator>
<dc:creator>Mayer-Blackwell, K.</dc:creator>
<dc:creator>Fiore-Gartland, A.</dc:creator>
<dc:creator>Minot, S.</dc:creator>
<dc:creator>Lin, P. J. C.</dc:creator>
<dc:creator>Tam, Y. K.</dc:creator>
<dc:creator>Weissman, D.</dc:creator>
<dc:creator>Kublin, J. G.</dc:creator>
<dc:date>2023-02-19</dc:date>
<dc:identifier>doi:10.1101/2023.02.18.529093</dc:identifier>
<dc:title><![CDATA[The Regulation of Nucleic Acid Vaccine Responses by the Microbiome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.20.529303v1?rss=1">
<title>
<![CDATA[
Validation of the association between MRI and gene signatures in facioscapulohumeral dystrophy muscle: implications for clinical trial design 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.20.529303v1?rss=1</link>
<description><![CDATA[
Identifying the aberrant expression of DUX4 in skeletal muscle as the cause of facioscapulohumeral dystrophy (FSHD) has led to rational therapeutic development and clinical trials. Several studies support the use of MRI characteristics and the expression of DUX4-regulated genes in muscle biopsies as biomarkers of FSHD disease activity and progression, but reproducibility across studies needs further validation. We performed lower-extremity MRI and muscle biopsies in the mid-portion of the tibialis anterior (TA) muscles bilaterally in FSHD subjects and validated our prior reports of the strong association between MRI characteristics and expression of genes regulated by DUX4 and other gene categories associated with FSHD disease activity. We further show that measurements of normalized fat content in the entire TA muscle strongly predict molecular signatures in the mid-portion of the TA. Together with moderate-to-strong correlations of gene signatures and MRI characteristics between the TA muscles bilaterally, these results suggest a whole muscle model of disease progression and provide a strong basis for inclusion of MRI and molecular biomarkers in clinical trial design.
]]></description>
<dc:creator>Wong, C.-J.</dc:creator>
<dc:creator>Friedman, S. D.</dc:creator>
<dc:creator>Snider, L.</dc:creator>
<dc:creator>Bennett, S. R.</dc:creator>
<dc:creator>Jones, T. I.</dc:creator>
<dc:creator>Jones, P. L.</dc:creator>
<dc:creator>Shaw, D. W.</dc:creator>
<dc:creator>Blemker, S. S.</dc:creator>
<dc:creator>Riem, L.</dc:creator>
<dc:creator>DuCharme, O.</dc:creator>
<dc:creator>Lemmers, R. J.</dc:creator>
<dc:creator>van der Maarel, S. M.</dc:creator>
<dc:creator>Wang, L. H.</dc:creator>
<dc:creator>Tawil, R.</dc:creator>
<dc:creator>Statland, J. M.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:date>2023-02-20</dc:date>
<dc:identifier>doi:10.1101/2023.02.20.529303</dc:identifier>
<dc:title><![CDATA[Validation of the association between MRI and gene signatures in facioscapulohumeral dystrophy muscle: implications for clinical trial design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529353v1?rss=1">
<title>
<![CDATA[
Multiplex Base Editing to Protect from CD33-Directed Therapy: Implications for Immune and Gene Therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529353v1?rss=1</link>
<description><![CDATA[
On-target toxicity to normal cells is a major safety concern with targeted immune and gene therapies. Here, we developed a base editing (BE) approach exploiting a naturally occurring CD33 single nucleotide polymorphism leading to removal of full-length CD33 surface expression on edited cells. CD33 editing in human and nonhuman primate (NHP) hematopoietic stem and progenitor cells (HSPCs) protects from CD33-targeted therapeutics without affecting normal hematopoiesis in vivo, thus demonstrating potential for novel immunotherapies with reduced off-leukemia toxicity. For broader applications to gene therapies, we demonstrated highly efficient (>70%) multiplexed adenine base editing of the CD33 and gamma globin genes, resulting in long-term persistence of dual gene-edited cells with HbF reactivation in NHPs. In vitro, dual gene-edited cells could be enriched via treatment with the CD33 antibody-drug conjugate, gemtuzumab ozogamicin (GO). Together, our results highlight the potential of adenine base editors for improved immune and gene therapies.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=85 SRC="FIGDIR/small/529353v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Borot, F.</dc:creator>
<dc:creator>Humbert, O.</dc:creator>
<dc:creator>Newby, G. A.</dc:creator>
<dc:creator>Fields, E.</dc:creator>
<dc:creator>Kohli, S.</dc:creator>
<dc:creator>Radtke, S.</dc:creator>
<dc:creator>Laszlo, G. S.</dc:creator>
<dc:creator>Mayuranathan, T.</dc:creator>
<dc:creator>Ali, A. M.</dc:creator>
<dc:creator>Weiss, M. J.</dc:creator>
<dc:creator>Yen, J. S.</dc:creator>
<dc:creator>Walter, R. B.</dc:creator>
<dc:creator>Liu, D. R.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Kiem, H.-P.</dc:creator>
<dc:date>2023-02-23</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529353</dc:identifier>
<dc:title><![CDATA[Multiplex Base Editing to Protect from CD33-Directed Therapy: Implications for Immune and Gene Therapy]]></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/10.1101/2023.02.24.529909v1?rss=1">
<title>
<![CDATA[
Chromosome-specific maturation of the epigenome in the Drosophila male germline 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.24.529909v1?rss=1</link>
<description><![CDATA[
Spermatogenesis in the Drosophila male germline proceeds through a unique transcriptional program controlled both by germline-specific transcription factors and by testis-specific versions of core transcriptional machinery. This program includes the activation of genes on the heterochromatic Y chromosome, and reduced transcription from the X chromosome, but how expression from these sex chromosomes is regulated has not been defined. To resolve this, we profiled active chromatin features in the testes from wildtype and meiotic arrest mutants and integrate this with single-cell gene expression data from the Fly Cell Atlas. These data assign the timing of promoter activation for genes with germline-enriched expression throughout spermatogenesis, and general alterations of promoter regulation in germline cells. By profiling both active RNA polymerase II and histone modifications in isolated spermatocytes, we detail widespread patterns associated with regulation of the sex chromosomes. Our results demonstrate that the X chromosome is not enriched for silencing histone modifications, implying that sex chromosome inactivation does not occur in the Drosophila male germline. Instead, a lack of dosage compensation in spermatocytes accounts for the reduced expression from this chromosome. Finally, profiling uncovers dramatic ubiquitinylation of histone H2A and lysine-16 acetylation of histone H4 across the Y chromosome in spermatocytes that may contribute to the activation of this heterochromatic chromosome.
]]></description>
<dc:creator>Anderson, J.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:date>2023-02-25</dc:date>
<dc:identifier>doi:10.1101/2023.02.24.529909</dc:identifier>
<dc:title><![CDATA[Chromosome-specific maturation of the epigenome in the Drosophila male germline]]></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/10.1101/2023.02.27.530304v1?rss=1">
<title>
<![CDATA[
Hypercholesterolemia aggravates in-stent restenosis in rabbits: a mitigating effect of stent surface modification with CD47-derived peptide. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.27.530304v1?rss=1</link>
<description><![CDATA[
BackgroundHypercholesterolemia (HC) has previously been shown to augment restenotic response in several animal models and humans. However, the mechanistic aspects of in-stent restenosis (ISR) on a hypercholesterolemic background, including potential augmentation of systemic and local inflammation precipitated by HC are not completely understood. CD47 is a transmembrane protein known to abort crucial inflammatory pathways. Our present studies have examined the interrelation between HC, inflammation, and ISR and investigated the therapeutic potential of stents coated with a CD47-derived peptide (pepCD47) in the hypercholesterolemic rabbit model.

Methods and ResultsPepCD47 was immobilized on metal foil coupons and stents using polybisphosphonate coordination chemistry and pyridyldithio/thiol conjugation. The relative abundance of the surface-associated cells on bare metal (BM) and pepCD47 foils exposed to whole rabbit blood showed a 40% inhibition of cell attachment on pepCD47-modified surfaces. Likewise, cytokine expression analyzed in buffy coat-derived cells cultured over the BM and pepCD47-derivatized foils demonstrated a M2/M1 increase with pepCD47 coating. Hypercholesterolemic and normocholesterolemic rabbit cohorts underwent bilateral implantation of BM and pepCD47 stents in the iliac location. Hypercholesterolemia increased neointimal growth in comparison with normocholesterolemic animals at 4 weeks post-stenting. These untoward outcomes were mitigated in the arteries of hypercholesterolemic rabbits treated with pepCD47-derivatized stents. Compared to NC animals, inflammatory cytokine immunopositivity and macrophage infiltration of peri-strut areas increased in HC group animals, and was attenuated in the arteries of hypercholesterolemic rabbits treated with pepCD47 stents.

ConclusionsAugmented inflammatory responses triggered by HC underlie severe ISR morphology in hypercholesterolemic rabbits. Blockage of initial platelet and leukocyte attachment to stent struts through CD47 functionalization of stents mitigates pro-restenotic effects of HC.
]]></description>
<dc:creator>Fishbein, I.</dc:creator>
<dc:creator>Inamdar, V. V.</dc:creator>
<dc:creator>Alferiev, I. S.</dc:creator>
<dc:creator>Bratinov, G.</dc:creator>
<dc:creator>Zviman, M. M.</dc:creator>
<dc:creator>Yekhilevsky, A.</dc:creator>
<dc:creator>Nagaswami, C.</dc:creator>
<dc:creator>Gardiner, K. L.</dc:creator>
<dc:creator>Levy, R. J.</dc:creator>
<dc:creator>Stachelek, S. J.</dc:creator>
<dc:date>2023-02-28</dc:date>
<dc:identifier>doi:10.1101/2023.02.27.530304</dc:identifier>
<dc:title><![CDATA[Hypercholesterolemia aggravates in-stent restenosis in rabbits: a mitigating effect of stent surface modification with CD47-derived peptide.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.28.530282v1?rss=1">
<title>
<![CDATA[
Olfactory and neuropeptide inputs to appetite neurons in the arcuate nucleus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.28.530282v1?rss=1</link>
<description><![CDATA[
The sense of smell has potent effects on appetite, but the underlying neural pathways remain undefined. Here we investigated how olfactory signals reach two subsets of appetite-linked neurons in the hypothalamic arcuate nucleus: AgRP (agouti-related peptide) neurons, which stimulate appetite, and POMC (pro-opiomelanocortin) neurons, which suppress it. Using polysynaptic viral tracing, we show that AgRP and POMC neurons receive indirect input from partially overlapping but distinct areas of the olfactory cortex, indicating that they process different sets of olfactory information. We also identify different complements of neurons directly upstream of AgRP and POMC neurons that can relay olfactory cortical signals to the appetite neurons. Single cell transcriptomics shows heterogeneous expression of neuromodulator receptors among AgRP neurons, suggesting variations in the signals they receive. Integrated viral tracing and RNA localization further reveals selected brain areas where upstream neurons express cognate receptor ligands. Together, these findings outline multiple pathways by which distinct olfactory and modulatory signals are differentially routed to neurons that promote versus inhibit appetite.
]]></description>
<dc:creator>Kuang, D.</dc:creator>
<dc:creator>Hanchate, N. K.</dc:creator>
<dc:creator>Lee, C.-Y.</dc:creator>
<dc:creator>Heck, A.</dc:creator>
<dc:creator>Ye, X.</dc:creator>
<dc:creator>Erdenebileg, M.</dc:creator>
<dc:creator>Buck, L. B.</dc:creator>
<dc:date>2023-03-01</dc:date>
<dc:identifier>doi:10.1101/2023.02.28.530282</dc:identifier>
<dc:title><![CDATA[Olfactory and neuropeptide inputs to appetite neurons in the arcuate nucleus]]></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/10.1101/2023.02.28.530516v1?rss=1">
<title>
<![CDATA[
Microbial community-scale metabolic modeling predicts personalized short-chain-fatty-acid production profiles in the human gut. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.28.530516v1?rss=1</link>
<description><![CDATA[
Microbially-derived short chain fatty acids (SCFAs) in the human gut are tightly coupled to host metabolism, immune regulation, and integrity of the intestinal epithelium. However, the production of SCFAs can vary widely between individuals consuming the same diet, with lower levels often associated with disease. A systems-scale mechanistic understanding of this heterogeneity is lacking. We present a microbial community-scale metabolic modeling (MCMM) approach to predict individual-specific SCFA production profiles. We assess the quantitative accuracy of our MCMMs using in vitro, ex vivo, and in vivo data. Next, we show how MCMM SCFA predictions are significantly associated with blood-derived clinical chemistries, including cardiometabolic and immunological health markers, across a large human cohort. Finally, we demonstrate how MCMMs can be leveraged to design personalized dietary, prebiotic, and probiotic interventions that optimize SCFA production in the gut. Our results represent an important advance in engineering gut microbiome functional outputs for precision health and nutrition.
]]></description>
<dc:creator>Bohmann, N.</dc:creator>
<dc:creator>Wilmanski, T.</dc:creator>
<dc:creator>Levy, L.</dc:creator>
<dc:creator>Lampe, J.</dc:creator>
<dc:creator>Gurry, T.</dc:creator>
<dc:creator>Rappaport, N.</dc:creator>
<dc:creator>Diener, C.</dc:creator>
<dc:creator>Gibbons, S. M.</dc:creator>
<dc:date>2023-03-01</dc:date>
<dc:identifier>doi:10.1101/2023.02.28.530516</dc:identifier>
<dc:title><![CDATA[Microbial community-scale metabolic modeling predicts personalized short-chain-fatty-acid production profiles in the human gut.]]></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/10.1101/2023.03.06.531408v1?rss=1">
<title>
<![CDATA[
Lethal giant larvae gene family (Llgl1 and Llgl2) functions as a tumor suppressor in mouse skin epidermis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.06.531408v1?rss=1</link>
<description><![CDATA[
Loss of cell polarity and tissue disorganization occurs in majority of epithelial cancers. Studies in simple model organisms identified molecular mechanisms responsible for the establishment and maintenance of cellular polarity, which play a pivotal role in establishing proper tissue architecture. The exact role of these cell polarity pathways in mammalian cancer is not completely understood. Here we analyzed the mammalian orthologs of drosophila apical-basal polarity gene lethal giant larvae (lgl), which regulates asymmetric stem cell division and functions as a tumor suppressor in flies. There are two mammalian orthologs of lgl (Llgl1 and Llgl2). To determine the role of the entire lgl signaling pathway in mammals we generated mice with ablation of both Llgl1 and Llgl2 in skin epidermis using K14-Cre (Llgl1/2-/- cKO mice). Surprisingly, we found that ablation of Llgl1/2 genes does not impact epidermal polarity in adult mice. However, old Llgl1/2 cKO mice present with focal skin lesions which are missing epidermal layer and ripe with inflammation. To determine the role of lgl signaling pathway in cancer we generated Trp53-/-/Llgl1/2-/- cKO and Trp53-/+/Llgl1/2-/- cKO mice. Loss of Llgl1/2 promoted squamous cell carcinoma (SCC) development in Trp53-/- cKO and caused SCC in Trp53-/+ cKO mice, while no cancer was observed in Trp53-/+ cKO controls. Mechanistically, we show that ablation of Llgl1/2 causes activation of aPKC and upregulation of NF-kB signaling pathway, which may be necessary for SCC in Trp53-/+/Llgl1/2-/- cKO mice. We conclude that Lgl signaling pathway functions as a tumor suppressor in mammalian skin epidermis.
]]></description>
<dc:creator>Bii, V. M.</dc:creator>
<dc:creator>Rudoy, D.</dc:creator>
<dc:creator>Klezovitch, O.</dc:creator>
<dc:creator>Vasioukhin, V.</dc:creator>
<dc:date>2023-03-08</dc:date>
<dc:identifier>doi:10.1101/2023.03.06.531408</dc:identifier>
<dc:title><![CDATA[Lethal giant larvae gene family (Llgl1 and Llgl2) functions as a tumor suppressor in mouse skin epidermis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.09.531940v1?rss=1">
<title>
<![CDATA[
Diet-induced glial insulin resistance impairs the clearance of neuronal debris 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.09.531940v1?rss=1</link>
<description><![CDATA[
Obesity significantly increases the risk of developing neurodegenerative disorders, yet the precise mechanisms underlying this connection remain unclear. Defects in glial phagocytic function are a key feature of neurodegenerative disorders, as delayed clearance of neuronal debris can result in inflammation, neuronal death, and poor nervous system recovery. Mounting evidence indicates that glial function can affect feeding behavior, weight, and systemic metabolism, suggesting that diet may play a role in regulating glial function. While it is appreciated that glial cells are insulin sensitive, whether obesogenic diets can induce glial insulin resistance and thereby impair glial phagocytic function remains unknown. Here, using a Drosophila model, we show that a chronic obesogenic diet induces glial insulin resistance and impairs the clearance of neuronal debris. Specifically, obesogenic diet exposure downregulates the basal and injury-induced expression of the glia-associated phagocytic receptor, Draper. Constitutive activation of systemic insulin release from Drosophila Insulin-producing cells (IPCs) mimics the effect of diet-induced obesity on glial draper expression. In contrast, genetically attenuating systemic insulin release from the IPCs rescues diet-induced glial insulin resistance and draper expression. Significantly, we show that genetically stimulating Phosphoinositide 3-kinase (PI3K), a downstream effector of Insulin receptor signaling, rescues HSD-induced glial defects. Hence, we establish that obesogenic diets impair glial phagocytic function and delays the clearance of neuronal debris.
]]></description>
<dc:creator>Alassaf, M.</dc:creator>
<dc:creator>Rajan, A.</dc:creator>
<dc:date>2023-03-10</dc:date>
<dc:identifier>doi:10.1101/2023.03.09.531940</dc:identifier>
<dc:title><![CDATA[Diet-induced glial insulin resistance impairs the clearance of neuronal debris]]></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/10.1101/2023.03.18.533255v1?rss=1">
<title>
<![CDATA[
Neuron cilia constrain glial regulators to microdomains around distal neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.18.533255v1?rss=1</link>
<description><![CDATA[
Each glia interacts with multiple neurons, but the fundamental logic of whether it interacts with all equally remains unclear. We find that a single sense-organ glia modulates different contacting neurons distinctly. To do so, it partitions regulatory cues into molecular microdomains at specific neuron contact-sites, at its delimited apical membrane. For one glial cue, K/Cl transporter KCC-3, microdomain-localization occurs through a two-step, neuron-dependent process. First, KCC-3 shuttles to glial apical membranes. Second, some contacting neuron cilia repel it, rendering it microdomain-localized around one distal neuron-ending. KCC-3 localization tracks animal aging, and while apical localization is sufficient for contacting neuron function, microdomain-restriction is required for distal neuron properties. Finally, we find the glia regulates its microdomains largely independently. Together, this uncovers that glia modulate cross-modal sensor processing by compartmentalizing regulatory cues into microdomains. Glia across species contact multiple neurons and localize disease-relevant cues like KCC-3. Thus, analogous compartmentalization may broadly drive how glia regulate information processing across neural circuits.
]]></description>
<dc:creator>Ray, S.</dc:creator>
<dc:creator>Gurung, P.</dc:creator>
<dc:creator>Manning, R. S.</dc:creator>
<dc:creator>Kravchuk, A.</dc:creator>
<dc:creator>Singhvi, A.</dc:creator>
<dc:date>2023-03-19</dc:date>
<dc:identifier>doi:10.1101/2023.03.18.533255</dc:identifier>
<dc:title><![CDATA[Neuron cilia constrain glial regulators to microdomains around distal neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.19.533377v1?rss=1">
<title>
<![CDATA[
Quantification of elongation stalls and impact on gene expression in yeast 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.19.533377v1?rss=1</link>
<description><![CDATA[
Ribosomal pauses are a critical part of co-translational events including protein folding and localization. However, extended ribosome pauses can lead to ribosome collisions, resulting in the activation of ribosome rescue pathways and turnover of protein and mRNA. While this relationship has been known, the specific threshold between permissible pausing versus activation of rescue pathways has not been quantified. We have taken a method used to measure elongation time and adapted it for use in S. cerevisiae to quantify the impact of elongation stalls. We find, in transcripts containing Arg CGA codon repeat-induced stalls, a Hel2-mediated dose-dependent decrease in protein expression and mRNA level and an elongation delay on the order of minutes. In transcripts that contain synonymous substitutions to non-optimal Leu codons, there is a decrease in protein and mRNA levels, as well as similar elongation delay, but this occurs through a non-Hel2-mediated mechanism. Finally, we find that Dhh1 selectively increases protein expression, mRNA level, and elongation rate. This indicates that distinct poorly translated codons in an mRNA will activate different rescue pathways despite similar elongation stall durations. Taken together, these results provide new quantitative mechanistic insight into the surveillance of translation and the roles of Hel2 and Dhh1 in mediating ribosome pausing events.
]]></description>
<dc:creator>Hou, W.</dc:creator>
<dc:creator>Harjono, V.</dc:creator>
<dc:creator>Harvey, A. T.</dc:creator>
<dc:creator>Subramaniam, A. R.</dc:creator>
<dc:creator>Zid, B. M.</dc:creator>
<dc:date>2023-03-20</dc:date>
<dc:identifier>doi:10.1101/2023.03.19.533377</dc:identifier>
<dc:title><![CDATA[Quantification of elongation stalls and impact on gene expression in yeast]]></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/10.1101/2023.03.21.533668v1?rss=1">
<title>
<![CDATA[
Molecular heterogeneity of C. elegans glia across sexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.21.533668v1?rss=1</link>
<description><![CDATA[
A comprehensive description of nervous system function, and sex dimorphism within, is incomplete without clear assessment of the diversity of its component cell types, neurons and glia. C. elegans has an invariant nervous system with the first mapped connectome of a multi- cellular organism and single-cell atlas of component neurons. Here we present single nuclear RNA-seq evaluation of glia across the entire adult C. elegans nervous system, including both sexes. Machine learning models enabled us to identify both sex-shared and sex-specific glia and glial subclasses. We have identified and validated molecular markers in silico and in vivo for these molecular subcategories. Comparative analytics also reveals previously unappreciated molecular heterogeneity in anatomically identical glia between and within sexes, indicating consequent functional heterogeneity. Furthermore, our datasets reveal that while adult C. elegans glia express neuropeptide genes, they lack the canonical unc-31/CAPS-dependent dense core vesicle release machinery. Thus, glia employ alternate neuromodulator processing mechanisms. Overall, this molecular atlas, available at www.wormglia.org, reveals rich insights into heterogeneity and sex dimorphism in glia across the entire nervous system of an adult animal.
]]></description>
<dc:creator>Purice, M. D.</dc:creator>
<dc:creator>Quitevis, E. J. A.</dc:creator>
<dc:creator>Manning, R. S.</dc:creator>
<dc:creator>Severs, L. J.</dc:creator>
<dc:creator>Tran, N. T.</dc:creator>
<dc:creator>Sorrentino, V.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:creator>Singhvi, A.</dc:creator>
<dc:date>2023-03-24</dc:date>
<dc:identifier>doi:10.1101/2023.03.21.533668</dc:identifier>
<dc:title><![CDATA[Molecular heterogeneity of C. elegans glia across sexes]]></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/10.1101/2023.03.23.533993v1?rss=1">
<title>
<![CDATA[
Mapping the neutralizing specificity of human anti-HIV serum by deep mutational scanning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.23.533993v1?rss=1</link>
<description><![CDATA[
Understanding the specificities of human serum antibodies that broadly neutralize HIV can inform prevention and treatment strategies. Here we describe a deep mutational scanning system that can measure the effects of combinations of mutations to HIV envelope (Env) on neutralization by antibodies and polyclonal serum. We first show that this system can accurately map how all functionally tolerated mutations to Env affect neutralization by monoclonal antibodies. We then comprehensively map Env mutations that affect neutralization by a set of human polyclonal sera known to target the CD4-binding site that neutralize diverse strains of HIV. The neutralizing activities of these sera target different epitopes, with most sera having specificities reminiscent of individual characterized monoclonal antibodies, but one sera targeting two epitopes within the CD4 binding site. Mapping the specificity of the neutralizing activity in polyclonal human serum will aid in assessing anti-HIV immune responses to inform prevention strategies.
]]></description>
<dc:creator>Radford, C. E.</dc:creator>
<dc:creator>Schommers, P.</dc:creator>
<dc:creator>Gieselmann, L.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Yu, T. C.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Klein, F.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2023-03-24</dc:date>
<dc:identifier>doi:10.1101/2023.03.23.533993</dc:identifier>
<dc:title><![CDATA[Mapping the neutralizing specificity of human anti-HIV serum by deep mutational scanning]]></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/10.1101/2023.04.04.535599v1?rss=1">
<title>
<![CDATA[
Cadherin-mediated cell-cell adhesion regulates collective pediatric glioma cell migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.04.535599v1?rss=1</link>
<description><![CDATA[
Pediatric high-grade gliomas are highly invasive and essentially incurable. Glioma cells migrate between neurons and glia, along axon tracts, and through extracellular matrix surrounding blood vessels and underlying the pia. Mechanisms that allow adaptation to such complex environments are poorly understood. N-cadherin is highly expressed in pediatric gliomas and associated with shorter survival. We found that inter-cellular homotypic N-cadherin interactions differentially regulate glioma migration according to the microenvironment, stimulating migration on cultured neurons or astrocytes but inhibiting invasion into reconstituted or astrocyte-deposited extracellular matrix. N-cadherin localizes to filamentous connections between migrating leader cells but to epithelial-like junctions between followers. Leader cells have more surface and recycling N-cadherin, increased YAP1/TAZ signaling, and increased proliferation relative to followers. YAP1/TAZ signaling is dynamically regulated as leaders and followers change position, leading to altered N-cadherin levels and organization. Together, the results suggest that pediatric glioma cells adapt to different microenvironments by regulating N-cadherin dynamics and cell-cell contacts.

SummaryPediatric gliomas invade the brain by migrating between nerve cells or exploiting extracellular matrix along blood vessels. This research reveals cross-talk between YAP1/TAZ signaling and N-cadherin that regulates leader-follower cell phenotypes and migration efficiency in neural and extracellular matrix environments.
]]></description>
<dc:creator>Kim, D.</dc:creator>
<dc:creator>Olson, J. M.</dc:creator>
<dc:creator>Cooper, J. A.</dc:creator>
<dc:date>2023-04-05</dc:date>
<dc:identifier>doi:10.1101/2023.04.04.535599</dc:identifier>
<dc:title><![CDATA[Cadherin-mediated cell-cell adhesion regulates collective pediatric glioma cell migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.05.535726v1?rss=1">
<title>
<![CDATA[
A single-cell transcriptional timelapse of mouse embryonic development, from gastrula to pup 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.05.535726v1?rss=1</link>
<description><![CDATA[
The house mouse, Mus musculus, is an exceptional model system, combining genetic tractability with close homology to human biology. Gestation in mouse development lasts just under three weeks, a period during which its genome orchestrates the astonishing transformation of a single cell zygote into a free-living pup composed of >500 million cells. Towards a global framework for exploring mammalian development, we applied single cell combinatorial indexing (sci-*) to profile the transcriptional states of 12.4 million nuclei from 83 precisely staged embryos spanning late gastrulation (embryonic day 8 or E8) to birth (postnatal day 0 or P0), with 2-hr temporal resolution during somitogenesis, 6-hr resolution through to birth, and 20-min resolution during the immediate postpartum period. From these data (E8 to P0), we annotate dozens of trajectories and hundreds of cell types and perform deeper analyses of the unfolding of the posterior embryo during somitogenesis as well as the ontogenesis of the kidney, mesenchyme, retina, and early neurons. Finally, we leverage the depth and temporal resolution of these whole embryo snapshots, together with other published data, to construct and curate a rooted tree of cell type relationships that spans mouse development from zygote to pup. Throughout this tree, we systematically nominate sets of transcription factors (TFs) and other genes as candidate drivers of the in vivo differentiation of hundreds of mammalian cell types. Remarkably, the most dramatic shifts in transcriptional state are observed in a restricted set of cell types in the hours immediately following birth, and presumably underlie the massive changes in physiology that must accompany the successful transition of a placental mammal to extrauterine life.
]]></description>
<dc:creator>Qiu, C.</dc:creator>
<dc:creator>Martin, B. K.</dc:creator>
<dc:creator>Welsh, I. C.</dc:creator>
<dc:creator>Daza, R. M.</dc:creator>
<dc:creator>Le, T.-M.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Nichols, E. K.</dc:creator>
<dc:creator>Taylor, M. L.</dc:creator>
<dc:creator>Fulton, O.</dc:creator>
<dc:creator>O'Day, D. R.</dc:creator>
<dc:creator>Gomes, A. R.</dc:creator>
<dc:creator>Ilcisin, S.</dc:creator>
<dc:creator>Srivatsan, S.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:creator>Hamazaki, N.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:creator>Kimelman, D.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Schier, A. F.</dc:creator>
<dc:creator>Spielmann, M.</dc:creator>
<dc:creator>Murray, S. A.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:date>2023-04-05</dc:date>
<dc:identifier>doi:10.1101/2023.04.05.535726</dc:identifier>
<dc:title><![CDATA[A single-cell transcriptional timelapse of mouse embryonic development, from gastrula to pup]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.08.536123v1?rss=1">
<title>
<![CDATA[
Antibodies that neutralize all current SARS-CoV-2 variants of concern by conformational locking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.08.536123v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 continues to evolve and evade most existing neutralizing antibodies, including all clinically authorized antibodies. We have isolated and characterized two human monoclonal antibodies, 12-16 and 12-19, which exhibited neutralizing activities against all SARS-CoV-2 variants tested, including BQ.1.1 and XBB.1.5. They also blocked infection in hamsters challenged with Omicron BA.1 intranasally. Structural analyses revealed both antibodies targeted a conserved quaternary epitope located at the interface between the N-terminal domain and subdomain 1, revealing a previously unrecognized site of vulnerability on SARS-CoV-2 spike. These antibodies prevent viral receptor engagement by locking the receptor-binding domain of spike in the down conformation, revealing a novel mechanism of virus neutralization for non-RBD antibodies. Deep mutational scanning showed that SARS-CoV-2 could mutate to escape 12-19, but the responsible mutations are rarely found in circulating viruses. Antibodies 12-16 and 12-19 hold promise as prophylactic agents for immunocompromised persons who do not respond robustly to COVID-19 vaccines.
]]></description>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Casner, R. G.</dc:creator>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Iketani, S.</dc:creator>
<dc:creator>Chan, J. F.-W.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Nair, M. S.</dc:creator>
<dc:creator>Mohri, H.</dc:creator>
<dc:creator>Reddem, E. R.</dc:creator>
<dc:creator>Yuan, S.</dc:creator>
<dc:creator>Poon, V. K.-M.</dc:creator>
<dc:creator>Chan, C. C.-S.</dc:creator>
<dc:creator>Yuen, K.-Y.</dc:creator>
<dc:creator>Sheng, Z.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Shapiro, L.</dc:creator>
<dc:creator>Ho, D. D.</dc:creator>
<dc:date>2023-04-10</dc:date>
<dc:identifier>doi:10.1101/2023.04.08.536123</dc:identifier>
<dc:title><![CDATA[Antibodies that neutralize all current SARS-CoV-2 variants of concern by conformational locking]]></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/10.1101/2023.04.09.536175v1?rss=1">
<title>
<![CDATA[
Single B cell transcriptomics identifies multiple isotypes of broadly neutralizing antibodies against flaviviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.09.536175v1?rss=1</link>
<description><![CDATA[
Sequential dengue virus (DENV) infections often generate neutralizing antibodies against all four DENV serotypes and sometimes, Zika virus. Characterizing cross-flavivirus broadly neutralizing antibody (bnAb) responses can inform countermeasure strategies that avoid infection enhancement associated with non-neutralizing antibodies. Here, we used single cell transcriptomics to mine the bnAb repertoire following secondary DENV infection. We identified several new bnAbs with comparable or superior breadth and potency to known bnAbs, and with distinct recognition determinants. Unlike all known flavivirus bnAbs, which are IgG1, one newly identified cross-flavivirus bnAb (F25.S02) was derived from IgA1. Both IgG1 and IgA1 versions of F25.S02 and known bnAbs displayed neutralizing activity, but only IgG1 enhanced infection in monocytes expressing IgG and IgA Fc receptors. Moreover, IgG-mediated enhancement of infection was inhibited by IgA1 versions of bnAbs. We demonstrate a role for IgA in flavivirus infection and immunity with implications for vaccine and therapeutic strategies.
]]></description>
<dc:creator>Lubow, J.</dc:creator>
<dc:creator>Levoir, L. M.</dc:creator>
<dc:creator>Ralph, D. K.</dc:creator>
<dc:creator>Belmont, L.</dc:creator>
<dc:creator>Contreras, M.</dc:creator>
<dc:creator>Cartwright-Acar, C. H.</dc:creator>
<dc:creator>Kikawa, C.</dc:creator>
<dc:creator>Kannan, S.</dc:creator>
<dc:creator>Davidson, E.</dc:creator>
<dc:creator>Doranz, B. J.</dc:creator>
<dc:creator>Duran, V.</dc:creator>
<dc:creator>Sanchez, D. E.</dc:creator>
<dc:creator>Sanz, A. M.</dc:creator>
<dc:creator>Rosso, F.</dc:creator>
<dc:creator>Einav, S.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Goo, L.</dc:creator>
<dc:date>2023-04-11</dc:date>
<dc:identifier>doi:10.1101/2023.04.09.536175</dc:identifier>
<dc:title><![CDATA[Single B cell transcriptomics identifies multiple isotypes of broadly neutralizing antibodies against flaviviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.10.536252v1?rss=1">
<title>
<![CDATA[
Estimating heritability explained by local ancestry and evaluating stratification bias in admixture mapping from summary statistics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.10.536252v1?rss=1</link>
<description><![CDATA[
The heritability explained by local ancestry markers in an admixed population [Formula] provides crucial insight into the genetic architecture of a complex disease or trait. Estimation of [Formula] can be susceptible to biases due to population structure in ancestral populations. Here, we present a novel approach, Heritability estimation from Admixture Mapping Summary STAtistics (HAMSTA), which uses summary statistics from admixture mapping to infer heritability explained by local ancestry while adjusting for biases due to ancestral stratification. Through extensive simulations, we demonstrate that HAMSTA [Formula] estimates are approximately unbiased and are robust to ancestral stratification compared to existing approaches. In the presence of ancestral stratification, we show a HAMSTA-derived sampling scheme provides a calibrated family-wise error rate (FWER) of [~]5% for admixture mapping, unlike existing FWER estimation approaches. We apply HAMSTA to 20 quantitative phenotypes of up to 15,988 self-reported African American individuals in the Population Architecture using Genomics and Epidemiology (PAGE) study. We observe [Formula] in the 20 phenotypes range from 0.0025 to 0.033 (mean [Formula]), which translates to [Formula] ranging from 0.062 to 0.85 (mean [Formula]). Across these phenotypes we find little evidence of inflation due to ancestral population stratification in current admixture mapping studies (mean inflation factor of 0.99 +/-0.001). Overall, HAMSTA provides a fast and powerful approach to estimate genome-wide heritability and evaluate biases in test statistics of admixture mapping studies.
]]></description>
<dc:creator>Chan, T. F.</dc:creator>
<dc:creator>Rui, X.</dc:creator>
<dc:creator>Conti, D. V.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Graff, M.</dc:creator>
<dc:creator>Haessler, J.</dc:creator>
<dc:creator>Haiman, C.</dc:creator>
<dc:creator>Highland, H. M.</dc:creator>
<dc:creator>Jung, S. Y.</dc:creator>
<dc:creator>Kenny, E.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Le Marchland, L.</dc:creator>
<dc:creator>North, K. E.</dc:creator>
<dc:creator>Tao, R.</dc:creator>
<dc:creator>Wojcik, G.</dc:creator>
<dc:creator>Gignoux, C. R.</dc:creator>
<dc:creator>PAGE Consortium,</dc:creator>
<dc:creator>Chiang, C. W.</dc:creator>
<dc:creator>Mancuso, N.</dc:creator>
<dc:date>2023-04-10</dc:date>
<dc:identifier>doi:10.1101/2023.04.10.536252</dc:identifier>
<dc:title><![CDATA[Estimating heritability explained by local ancestry and evaluating stratification bias in admixture mapping from summary statistics]]></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/10.1101/2023.04.12.536510v1?rss=1">
<title>
<![CDATA[
ME-Bayes SL: Enhanced Bayesian Polygenic Risk Prediction Leveraging Information across Multiple Ancestry Groups 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.12.536510v1?rss=1</link>
<description><![CDATA[
Polygenic risk scores (PRS) are now showing promising predictive performance on a wide variety of complex traits and diseases, but there exists a substantial performance gap across different populations. We propose MUSSEL, a method for ancestry-specific polygenic prediction that borrows information in the summary statistics from genome-wide association studies (GWAS) across multiple ancestry groups. MUSSEL conducts Bayesian hierarchical modeling under a MUltivariate Spike-and-Slab model for effect-size distribution and incorporates an Ensemble Learning step using super learner to combine information across different tuning parameter settings and ancestry groups. In our simulation studies and data analyses of 16 traits across four distinct studies, totaling 5.7 million participants with a substantial ancestral diversity, MUSSEL shows promising performance compared to alternatives. The method, for example, has an average gain in prediction R2 across 11 continuous traits of 40.2% and 49.3% compared to PRS-CSx and CT-SLEB, respectively, in the African Ancestry population. The best-performing method, however, varies by GWAS sample size, target ancestry, underlying trait architecture, and the choice of reference samples for LD estimation, and thus ultimately, a combination of methods may be needed to generate the most robust PRS across diverse populations.
]]></description>
<dc:creator>Jin, J.</dc:creator>
<dc:creator>Zhan, J.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Zhao, R.</dc:creator>
<dc:creator>O'Connell, J.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>23andMe Research Team,</dc:creator>
<dc:creator>Buyske, S.</dc:creator>
<dc:creator>Gignoux, C. R.</dc:creator>
<dc:creator>Haiman, C. A.</dc:creator>
<dc:creator>Kenny, E.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>North, K. E.</dc:creator>
<dc:creator>Koelsch, B. L.</dc:creator>
<dc:creator>Wojcik, G. L.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Chatterjee, N.</dc:creator>
<dc:date>2023-04-13</dc:date>
<dc:identifier>doi:10.1101/2023.04.12.536510</dc:identifier>
<dc:title><![CDATA[ME-Bayes SL: Enhanced Bayesian Polygenic Risk Prediction Leveraging Information across Multiple Ancestry Groups]]></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/10.1101/2023.04.12.536619v1?rss=1">
<title>
<![CDATA[
Carcinoma-associated fibroblast-like tumor cells remodel the Ewing sarcoma tumor microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.12.536619v1?rss=1</link>
<description><![CDATA[
Tumor heterogeneity is a major driver of cancer progression. In epithelial-derived malignancies, carcinoma-associated fibroblasts (CAFs) contribute to tumor heterogeneity by depositing extracellular matrix (ECM) proteins that dynamically remodel the tumor microenvironment (TME). Ewing sarcomas (EwS) are histologically monomorphous, mesenchyme-derived tumors that are devoid of CAFs. Here we identify a previously uncharacterized subpopulation of transcriptionally distinct EwS tumor cells that deposit pro-tumorigenic ECM. Single cell analyses revealed that these CAF-like cells differ from bulk EwS cells by their upregulation of a matrisome-rich gene signature that is normally repressed by EWS::FLI1, the oncogenic fusion transcription factor that underlies EwS pathogenesis. Further, our studies showed that ECM-depositing tumor cells express the cell surface marker CD73, allowing for their isolation ex vivo and detection in situ. Spatial profiling of tumor xenografts and patient biopsies demonstrated that CD73+ EwS cells and tumor cell-derived ECM are prevalent along tumor borders and invasive fronts. Importantly, despite loss of EWS::FLI1-mediated gene repression, CD73+ EwS cells retain expression of EWS::FLI1 and the fusion-activated gene signature, as well as tumorigenic and proliferative capacities. Thus, EwS tumor cells can be reprogrammed to adopt CAF-like properties and these transcriptionally and phenotypically distinct cell subpopulations contribute to tumor heterogeneity by remodeling the TME.
]]></description>
<dc:creator>Wrenn, E. D.</dc:creator>
<dc:creator>Apfelbaum, A. A.</dc:creator>
<dc:creator>Rudzinski, E. R.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Jiang, W.</dc:creator>
<dc:creator>Sud, S.</dc:creator>
<dc:creator>Van Noord, R. A.</dc:creator>
<dc:creator>Newman, E. A.</dc:creator>
<dc:creator>Garcia, N. M.</dc:creator>
<dc:creator>Hoglund, V. J.</dc:creator>
<dc:creator>Bhise, S. S.</dc:creator>
<dc:creator>Kanaan, S. B.</dc:creator>
<dc:creator>Waltner, O. G.</dc:creator>
<dc:creator>Furlan, S. N.</dc:creator>
<dc:creator>Lawlor, E. R.</dc:creator>
<dc:date>2023-04-13</dc:date>
<dc:identifier>doi:10.1101/2023.04.12.536619</dc:identifier>
<dc:title><![CDATA[Carcinoma-associated fibroblast-like tumor cells remodel the Ewing sarcoma tumor microenvironment]]></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/10.1101/2023.04.12.536652v1?rss=1">
<title>
<![CDATA[
Combinatorial genetic strategy accelerates the discovery of cancer genotype-phenotype associations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.12.536652v1?rss=1</link>
<description><![CDATA[
Available genetically-defined cancer models are limited in genotypic and phenotypic complexity and underrepresent the heterogeneity of human cancer. Herein, we describe a combinatorial genetic strategy applied to an organoid transformation assay to rapidly generate diverse, clinically relevant bladder and prostate cancer models. Importantly, the clonal architecture of the resultant tumors can be resolved using single-cell or spatially resolved next-generation sequencing to uncover polygenic drivers of cancer phenotypes.
]]></description>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Wong, A.</dc:creator>
<dc:creator>Sun, H.</dc:creator>
<dc:creator>Bhatia, V.</dc:creator>
<dc:creator>Javier, G.</dc:creator>
<dc:creator>Jana, S.</dc:creator>
<dc:creator>Mongomery, R. B.</dc:creator>
<dc:creator>Wright, J. L.</dc:creator>
<dc:creator>Lam, H.-M.</dc:creator>
<dc:creator>Hsieh, A. C.</dc:creator>
<dc:creator>Faltas, B. M.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:date>2023-04-14</dc:date>
<dc:identifier>doi:10.1101/2023.04.12.536652</dc:identifier>
<dc:title><![CDATA[Combinatorial genetic strategy accelerates the discovery of cancer genotype-phenotype associations]]></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/10.1101/2023.04.17.537247v1?rss=1">
<title>
<![CDATA[
Adenovirus protein VII binds the A-box of HMGB1 to repress interferon responses. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.17.537247v1?rss=1</link>
<description><![CDATA[
Viruses hijack host proteins to promote infection and dampen host defenses. Adenovirus encodes the multifunctional protein VII that serves both to compact viral genomes inside the virion and disrupt host chromatin. Protein VII binds the abundant nuclear protein high mobility group box 1 (HMGB1) and sequesters HMGB1 in chromatin. HMGB1 is an abundant host nuclear protein that can also be released from infected cells as an alarmin to amplify inflammatory responses. By sequestering HMGB1, protein VII prevents its release, thus inhibiting downstream inflammatory signaling. However, the consequences of this chromatin sequestration on host transcription are unknown. Here, we employ bacterial two-hybrid interaction assays and human cell biological systems to interrogate the mechanism of the protein VII-HMGB1 interaction. HMGB1 contains two DNA binding domains, the A- and B-boxes, that bend DNA to promote transcription factor binding while the C-terminal tail regulates this interaction. We demonstrate that protein VII interacts directly with the A-box of HMGB1, an interaction that is inhibited by the HMGB1 C-terminal tail. By cellular fractionation, we show that protein VII renders A-box containing constructs insoluble, thereby acting to prevent their release from cells. This sequestration is not dependent on HMGB1s ability to bind DNA but does require post-translational modifications on protein VII. Importantly, we demonstrate that protein VII inhibits expression of interferon {beta}, in an HMGB1- dependent manner, but does not affect transcription of downstream interferon- stimulated genes. Together, our results demonstrate that protein VII specifically harnesses HMGB1 through its A-box domain to depress the innate immune response and promote infection.
]]></description>
<dc:creator>Arnold, E. A.</dc:creator>
<dc:creator>Kaai, R. J.</dc:creator>
<dc:creator>Leung, K.</dc:creator>
<dc:creator>Brinkley, M. R.</dc:creator>
<dc:creator>Kelnhofer-Millevolte, L. E.</dc:creator>
<dc:creator>Guo, M. S.</dc:creator>
<dc:creator>Avgousti, D. C.</dc:creator>
<dc:date>2023-04-18</dc:date>
<dc:identifier>doi:10.1101/2023.04.17.537247</dc:identifier>
<dc:title><![CDATA[Adenovirus protein VII binds the A-box of HMGB1 to repress interferon responses.]]></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/10.1101/2023.04.22.537934v1?rss=1">
<title>
<![CDATA[
CDK7 is a Novel Therapeutic Vulnerability in Fibrolamellar Carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.22.537934v1?rss=1</link>
<description><![CDATA[
Fibrolamellar carcinoma (FLC) is a rare and lethal cancer that afflicts young individuals. The tumor arises in the background of a healthy liver, and patients typically present with advanced cancer at the time of diagnosis. Unfortunately, for these patients with advanced or recurrent cancer, no proven systemic therapies exist resulting in only 30-45% of patients surviving to 5 years. Investigations into the molecular underpinning of FLC have revealed a unique gene fusion between heat shock protein 40 (DNAJB1) and the catalytic subunit alpha of protein kinase A (PRKACA), leading to the formation of an oncoprotein (DNAJ-PKAc) that retains kinase activity and is a proven tumor-causing event in FLC. To uncover potential therapeutic targets, we engineered an FLC cell line by introducing the DNAJB1-PRKACA oncogene rearrangement into human hepatocellular cells using CRISPR/Cas9. We identified aberrant cell cycle progression, and follow-up molecular analysis revealed evidence of enhanced cyclin dependent kinase 7 (CDK7) activation in the DNAJB1-PRKACA expressing FLC cells. These findings were confirmed in human samples of FLC. In turn, targeting CDK7 with selective inhibitors demonstrated efficacy in several patient-derived models of FLC, with minimal toxicity to normal liver. Collectively, this work uncovers a novel candidate therapeutic vulnerability in FLC.
]]></description>
<dc:creator>Nukaya, M.</dc:creator>
<dc:creator>Cafferty, C.</dc:creator>
<dc:creator>Zahed, K.</dc:creator>
<dc:creator>Yun, I.</dc:creator>
<dc:creator>Al-Adra, D. P.</dc:creator>
<dc:creator>Kazim, N. A.</dc:creator>
<dc:creator>Farghli, A. R.</dc:creator>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Kratz, J. D.</dc:creator>
<dc:creator>Berres, M. E.</dc:creator>
<dc:creator>Yen, A.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:creator>Sethupathy, P.</dc:creator>
<dc:creator>Bradfield, C. A.</dc:creator>
<dc:creator>Ronnekleiv-Kelly, S. M.</dc:creator>
<dc:date>2023-04-23</dc:date>
<dc:identifier>doi:10.1101/2023.04.22.537934</dc:identifier>
<dc:title><![CDATA[CDK7 is a Novel Therapeutic Vulnerability in Fibrolamellar Carcinoma]]></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/10.1101/2023.04.24.538128v1?rss=1">
<title>
<![CDATA[
Identification of allele-specific KIV-2 repeats and impact on Lp(a) measurements for cardiovascular disease risk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.24.538128v1?rss=1</link>
<description><![CDATA[
The abundance of Lp(a) protein holds significant implications for the risk of cardiovascular disease (CVD), which is directly impacted by the copy number (CN) of KIV-2, a 5.5 kbp sub-region. KIV-2 is highly polymorphic in the population and accurate analysis is challenging. In this study, we present the DRAGEN KIV-2 CN caller, which utilizes short reads. Data across 166 WGS show that the caller has high accuracy, compared to optical mapping and can further phase [~]50% of the samples. We compared KIV-2 CN numbers to 24 previously postulated KIV-2 relevant SNVs, revealing that many are ineffective predictors of KIV-2 copy number. Population studies, including USA-based cohorts, showed distinct KIV-2 CN, distributions for European-, African-, and Hispanic-American populations and further underscored the limitations of SNV predictors. We demonstrate that the CN estimates correlate significantly with the available Lp(a) protein levels and that phasing is highly important.
]]></description>
<dc:creator>Behera, S.</dc:creator>
<dc:creator>Belyeu, J.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Paulin, L. F.</dc:creator>
<dc:creator>Nguyen, N. Q.</dc:creator>
<dc:creator>Newman, E.</dc:creator>
<dc:creator>Mahmoud, M.</dc:creator>
<dc:creator>Menon, V. K.</dc:creator>
<dc:creator>Qi, Q.</dc:creator>
<dc:creator>Joshi, P.</dc:creator>
<dc:creator>Marcovina, S.</dc:creator>
<dc:creator>Rossi, M.</dc:creator>
<dc:creator>Roller, E.</dc:creator>
<dc:creator>Han, J.</dc:creator>
<dc:creator>Onuchic, V.</dc:creator>
<dc:creator>Avery, C. L.</dc:creator>
<dc:creator>Ballantyne, C. M.</dc:creator>
<dc:creator>Rodriguez, C. J.</dc:creator>
<dc:creator>Kaplan, R. C.</dc:creator>
<dc:creator>Muzny, D. M.</dc:creator>
<dc:creator>Metcalf, G. A.</dc:creator>
<dc:creator>Gibbs, R.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Eberle, M. A.</dc:creator>
<dc:creator>Sedlazeck, F. J.</dc:creator>
<dc:date>2023-04-27</dc:date>
<dc:identifier>doi:10.1101/2023.04.24.538128</dc:identifier>
<dc:title><![CDATA[Identification of allele-specific KIV-2 repeats and impact on Lp(a) measurements for cardiovascular disease risk]]></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/10.1101/2023.05.04.539483v1?rss=1">
<title>
<![CDATA[
Machine vision reveals micronucleus rupture as a potential driver of the transcriptomic response to aneuploidy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.04.539483v1?rss=1</link>
<description><![CDATA[
Recent advances in isolating cells based on visual phenotypes have transformed our ability to identify the mechanisms and consequences of complex traits. Micronucleus (MN) formation is a frequent outcome of genome instability, triggers extensive changes in genome structure and signaling coincident with MN rupture, and is almost exclusively defined by visual analysis. Automated MN detection in microscopy images has proved challenging, limiting discovery of the mechanisms and consequences of MN. In this study we describe two new MN segmentation modules: a rapid model for classifying micronucleated cells and their rupture status (VCS MN), and a robust model for accurate MN segmentation (MNFinder) from a broad range of cell lines. As proof-of-concept, we define the transcriptome of non-transformed human cells with intact or ruptured MN after chromosome missegregation by combining VCS MN with photoactivation-based cell isolation and RNASeq. Surprisingly, we find that neither MN formation nor rupture triggers a strong unique transcriptional response. Instead, transcriptional changes appear correlated with small increases in aneuploidy in these cell classes. Our MN segmentation modules overcome a significant challenge with reproducible MN quantification, and, joined with visual cell sorting, enable the application of powerful functional genomics assays to a wide-range of questions in MN biology.
]]></description>
<dc:creator>DiPeso, L.</dc:creator>
<dc:creator>Pendyala, S.</dc:creator>
<dc:creator>Huang, H. Z.</dc:creator>
<dc:creator>Fowler, D. M.</dc:creator>
<dc:creator>Hatch, E. M.</dc:creator>
<dc:date>2023-05-05</dc:date>
<dc:identifier>doi:10.1101/2023.05.04.539483</dc:identifier>
<dc:title><![CDATA[Machine vision reveals micronucleus rupture as a potential driver of the transcriptomic response to aneuploidy]]></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/10.1101/2023.05.11.540440v1?rss=1">
<title>
<![CDATA[
Within-host rhinovirus evolution in upper and lower respiratory tract highlights capsid variability and mutation-independent compartmentalization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540440v1?rss=1</link>
<description><![CDATA[
BackgroundHuman rhinovirus (HRV) infections can progress from the upper (URT) to lower (LRT) respiratory tract in immunocompromised individuals, causing high rates of fatal pneumonia. Little is known about how HRV evolves within hosts during infection.

MethodsWe sequenced HRV complete genomes from 12 hematopoietic cell transplant patients with prolonged infection for up to 190 days from both URT (nasal wash, NW) and LRT (bronchoalveolar lavage, BAL) specimens. Metagenomic (mNGS) and amplicon-based NGS were used to study the emergence and evolution of intra-host single nucleotide variants (iSNVs).

ResultsIdentical HRV intra-host populations in matched NW and BAL specimens indicated no genetic adaptation is required for HRV to progress from URT to LRT. Microbial composition between matched NW and BAL confirmed no cross-contamination during sampling procedure. Coding iSNVs were 2.3-fold more prevalent in capsid over non-structural genes, adjusted for length. iSNVs modeled onto HRV capsid structures were significantly more likely to be found in surface residues, but were not preferentially located in known HRV neutralizing antibody epitopes. Newly emergent, serotype-matched iSNV haplotypes from immunocompromised individuals from 2008-2010 could be detected in Seattle-area community HRV sequences from 2020-2021.

ConclusionHRV infections in immunocompromised hosts can progress from URT to LRT with no specific evolutionary requirement. Capsid proteins carry the highest variability and emergent mutations can be detected in other, including future, HRV sequences.
]]></description>
<dc:creator>Makhsous, N.</dc:creator>
<dc:creator>Goya, S.</dc:creator>
<dc:creator>Avendano, C.</dc:creator>
<dc:creator>Rupp, J.</dc:creator>
<dc:creator>Kuypers, J.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Boeckh, M.</dc:creator>
<dc:creator>Waghmare, A.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:date>2023-05-11</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540440</dc:identifier>
<dc:title><![CDATA[Within-host rhinovirus evolution in upper and lower respiratory tract highlights capsid variability and mutation-independent compartmentalization]]></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/10.1101/2023.05.17.541189v1?rss=1">
<title>
<![CDATA[
Oncogenic reactivation of young L1s is a hallmark of colon cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.17.541189v1?rss=1</link>
<description><![CDATA[
Transposable elements become increasingly active in both cancerous and aging cells, driven by loss of DNA methylation as cells divide. Here we leverage the epigenomes of colon cancers with matched adjacent tissue, in addition to non-cancerous normals and cell line models, to assess the role of transposable elements as drivers or passengers in cancer development. Using the baseline of activity from normal and adjacent tissue, we show that the youngest subfamilies of the LINE1 (L1) family exhibit a degree of activity and recurrence across patients that goes beyond what is expected from hypomethylation and cell division, suggesting an additional mechanism of oncogenic reactivation. We characterize this mechanism and find that the loss of the tumor suppressor PLZF drives young L1 reactivation in a cell-division-independent manner. PLZF de-repression exposes abundant motifs for tumor core factors in the L1 5UTR. Active young L1s act as oncogenic enhancers, interacting with oncogenes via gained chromatin loops. We uncover oncogenic L1 reactivation as a hallmark of colon cancer, where young L1s activate universally in our cohort at high levels of recurrence, act as enhancers to oncogenes, and become wired into the core regulatory circuitry of colon cancer.
]]></description>
<dc:creator>Neu, D.</dc:creator>
<dc:creator>Hung, S.</dc:creator>
<dc:creator>Bartels, C. F.</dc:creator>
<dc:creator>Faber, Z. J.</dc:creator>
<dc:creator>Lovrenert, K.</dc:creator>
<dc:creator>Pontius, W. D.</dc:creator>
<dc:creator>Morgan, L.</dc:creator>
<dc:creator>Chakraborty, M.</dc:creator>
<dc:creator>Liao, W.</dc:creator>
<dc:creator>Chin, D.</dc:creator>
<dc:creator>Hong, E. S.</dc:creator>
<dc:creator>Gray, J.</dc:creator>
<dc:creator>Moreno, V.</dc:creator>
<dc:creator>Kalady, M.</dc:creator>
<dc:creator>Peters, U.</dc:creator>
<dc:creator>Gryder, B.</dc:creator>
<dc:creator>Sallari, R. C.</dc:creator>
<dc:creator>Scacheri, P. C.</dc:creator>
<dc:date>2023-05-21</dc:date>
<dc:identifier>doi:10.1101/2023.05.17.541189</dc:identifier>
<dc:title><![CDATA[Oncogenic reactivation of young L1s is a hallmark of colon cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.24.542133v1?rss=1">
<title>
<![CDATA[
Improving the study of RNA dynamics through advances in RNA-seq with metabolic labeling and nucleotide-recoding chemistry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.24.542133v1?rss=1</link>
<description><![CDATA[
RNA metabolic labeling using 4-thiouridine (s4U) captures the dynamics of RNA synthesis and decay. The power of this approach is dependent on appropriate quantification of labeled and unlabeled sequencing reads, which can be compromised by the apparent loss of s4U-labeled reads in a process we refer to as dropout. Here we show that s4U-containing transcripts can be selectively lost when RNA samples are handled under sub-optimal conditions, but that this loss can be minimized using an optimized protocol. We demonstrate a second cause of dropout in nucleotide recoding and RNA sequencing (NR-seq) experiments that is computational and downstream of library preparation. NR-seq experiments involve chemically converting s4U from a uridine analog to a cytidine analog and using the apparent T-to-C mutations to identify the populations of newly synthesized RNA. We show that high levels of T-to-C mutations can prevent read alignment with some computational pipelines, but that this bias can be overcome using improved alignment pipelines. Importantly, kinetic parameter estimates are affected by dropout independent of the NR chemistry employed, and all chemistries are practically indistinguishable in bulk, short-read RNA-seq experiments. Dropout is an avoidable problem that can be identified by including unlabeled controls, and mitigated through improved sample handing and read alignment that together improve the robustness and reproducibility of NR-seq experiments.
]]></description>
<dc:creator>Zimmer, J. T.</dc:creator>
<dc:creator>Vock, I. W.</dc:creator>
<dc:creator>Schofield, J. A.</dc:creator>
<dc:creator>Kiefer, L.</dc:creator>
<dc:creator>Moon, M. H.</dc:creator>
<dc:creator>Simon, M. D.</dc:creator>
<dc:date>2023-05-24</dc:date>
<dc:identifier>doi:10.1101/2023.05.24.542133</dc:identifier>
<dc:title><![CDATA[Improving the study of RNA dynamics through advances in RNA-seq with metabolic labeling and nucleotide-recoding chemistry]]></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/10.1101/2023.05.25.542327v1?rss=1">
<title>
<![CDATA[
Neural Network Models for Sequence-Based TCR and HLA Association Prediction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.25.542327v1?rss=1</link>
<description><![CDATA[
T cells rely on their T cell receptors (TCRs) to recognize foreign antigens presented by human leukocyte antigen (HLA) proteins. TCRs contain a record of an individuals past immune activities, and some TCRs are observed only in individuals with certain HLA alleles. As a result, characterising TCRs requires a thorough understanding of TCR-HLA associations. To this end, we propose a neural network method named Deep learning Prediction of TCR-HLA association (DePTH) to predict TCR-HLA associations based on their amino acid sequences. We show that the DePTH can be used to quantify the functional similarities of HLA alleles, and that these HLA similarities are associated with the survival outcomes of cancer patients who received immune checkpoint blockade treatment.
]]></description>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2023-05-26</dc:date>
<dc:identifier>doi:10.1101/2023.05.25.542327</dc:identifier>
<dc:title><![CDATA[Neural Network Models for Sequence-Based TCR and HLA Association Prediction]]></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/10.1101/2023.05.28.542640v1?rss=1">
<title>
<![CDATA[
Tracking the Metabolic Fate of Exogenous Arachidonic Acid in Ferroptosis Using Dual-Isotope Labeling Lipidomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.28.542640v1?rss=1</link>
<description><![CDATA[
Lipid metabolism is implicated in a variety of diseases, including cancer, cell death, and inflammation, but lipidomics has proven to be challenging due to the vast structural diversity over a narrow range of mass and polarity of lipids. Isotope labeling is often used in metabolomics studies to follow the metabolism of exogenously added labeled compounds because they can be differentiated from endogenous compounds by the mass shift associated with the label. The application of isotope labeling to lipidomics has also been explored as a method to track the metabolism of lipids in various disease states. However, it can be difficult to differentiate a single isotopically labeled lipid from the rest of the lipidome due to the variety of endogenous lipids present over the same mass range. Here we report the development of a dual-isotope deuterium labeling method to track the metabolic fate of exogenous polyunsaturated fatty acids, e.g, arachidonic acid (AA), in the context of ferroptosis using hydrophilic interaction-ion mobility-mass spectrometry (HILIC-IM-MS). Ferroptosis is a type of cell death that is dependent on lipid peroxidation. The use of two isotope labels rather than one enables the identification of labeled species by a signature doublet peak in the resulting mass spectra. A Python-based software, D-Tracer, was developed to efficiently extract metabolites with dual-isotope labels. The labeled species were then identified with Lipydomics based on their retention times, collision cross section, and m/z values. Changes in exogenous AA incorporation in the absence and presence of a ferroptosis inducer were elucidated.

Table of Contents

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]]></description>
<dc:creator>Reimers, N.</dc:creator>
<dc:creator>Do, Q.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Guo, A.</dc:creator>
<dc:creator>Ostrander, R.</dc:creator>
<dc:creator>Shoji, A. T.</dc:creator>
<dc:creator>Vuong, C.</dc:creator>
<dc:creator>Xu, L.</dc:creator>
<dc:date>2023-05-28</dc:date>
<dc:identifier>doi:10.1101/2023.05.28.542640</dc:identifier>
<dc:title><![CDATA[Tracking the Metabolic Fate of Exogenous Arachidonic Acid in Ferroptosis Using Dual-Isotope Labeling Lipidomics]]></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/10.1101/2023.05.30.542951v1?rss=1">
<title>
<![CDATA[
A single polymorphic residue in humans underlies species-specific restriction of HSV-1 by the antiviral protein MxB 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.30.542951v1?rss=1</link>
<description><![CDATA[
Myxovirus resistance proteins (MxA and MxB) are interferon-induced proteins that exert antiviral activity against a diverse range of RNA and DNA viruses. In primates, MxA has been shown to inhibit myxoviruses, bunyaviruses, and hepatitis B virus, whereas MxB restricts retroviruses and herpesviruses. As a result of their conflicts with viruses, both genes have been undergoing diversifying selection during primate evolution. Here, we investigate how MxB evolution in primates has affected its restriction of herpesviruses. In contrast to human MxB, we find that most primate orthologs, including the closely related chimpanzee MxB, do not inhibit HSV-1 replication. However, all primate MxB orthologs tested restrict human cytomegalovirus. Through the generation of human and chimpanzee MxB chimeras we show that a single residue, M83, is the key determinant of restriction of HSV-1 replication. Humans are the only primate species known to encode a methionine at this position, whereas most other primate species encode a lysine. Residue 83 is also the most polymorphic residue in MxB in human populations, with M83 being the most common variant. However, [~]2.5% of human MxB alleles encode a threonine at this position, which does not restrict HSV-1. Thus, a single amino acid variant in MxB, which has recently risen to high frequency in humans, has endowed humans with HSV-1 antiviral activity.

ImportanceHerpesviruses present a major global disease burden. Understanding the host cell mechanisms that block viral infections as well as how viruses can evolve to counteract these host defenses is critically important for understanding viral disease pathogenesis, and for developing therapeutic tools aimed at treating or preventing viral infections. Additionally, understanding how these host and viral mechanisms adapt to counter one another can aid in identifying the risks of, and barriers to, cross-species transmission events. As highlighted by the recent SARS-CoV-2 pandemic, episodic transmission events can have severe consequences for human health. This study reveals that the major human variant of the antiviral protein MxB inhibits the human pathogen HSV-1, whereas human minor variants and orthologous MxB genes from even closely related primates do not. Thus, in contrast to the many antagonistic virus-host interactions in which the virus is successful in thwarting the defense systems of their native hosts, in this case the human gene appears to be at least temporarily winning at this interface of the primate-herpesviral evolutionary arms race. Our findings further show that a polymorphism at amino acid 83 in a small fraction of the human population is sufficient to abrogate MxBs ability to inhibit HSV-1, which could have important implications for human susceptibility to HSV-1 pathogenesis.
]]></description>
<dc:creator>Bayer, A.</dc:creator>
<dc:creator>Child, S. J.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:creator>Geballe, A. P.</dc:creator>
<dc:date>2023-05-30</dc:date>
<dc:identifier>doi:10.1101/2023.05.30.542951</dc:identifier>
<dc:title><![CDATA[A single polymorphic residue in humans underlies species-specific restriction of HSV-1 by the antiviral protein MxB]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/265082v1?rss=1">
<title>
<![CDATA[
A Highly Efficient and Faithful MDS Patient-Derived Xenotransplantation Model for Pre-Clinical Studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/265082v1?rss=1</link>
<description><![CDATA[
Comprehensive preclinical studies of Myelodysplastic Syndromes (MDS) have been elusive due to limited ability of MDS stem cells to engraft current immunodeficient murine hosts. We developed a novel MDS patient-derived xenotransplantation model in cytokine-humanized immunodeficient "MISTRG" mice that for the first time provides efficient and faithful disease representation across all MDS subtypes. MISTRG MDS patient-derived xenografts (PDX) reproduce patients' dysplastic morphology with multi-lineage representation, including erythro- and megakaryopoiesis. MISTRG MDS-PDX replicate the original sample's genetic complexity and can be propagated via serial transplantation. MISTRG MDS-PDX demonstrate the cytotoxic and differentiation potential of targeted therapeutics providing superior readouts of drug mechanism of action and therapeutic efficacy. Physiologic humanization of the hematopoietic stem cell niche proves critical to MDS stem cell propagation and function in vivo. The MISTRG MDS-PDX model opens novel avenues of research and long-awaited opportunities in MDS research.
]]></description>
<dc:creator>Song, Y.</dc:creator>
<dc:creator>Rongvaux, A.</dc:creator>
<dc:creator>Taylor, A.</dc:creator>
<dc:creator>Jiang, T.</dc:creator>
<dc:creator>Tebaldi, T.</dc:creator>
<dc:creator>Balasubramanian, K.</dc:creator>
<dc:creator>Bagale, A.</dc:creator>
<dc:creator>Terzi, Y. K.</dc:creator>
<dc:creator>Gbyli, R.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Podoltsev, N.</dc:creator>
<dc:creator>Xu, M.</dc:creator>
<dc:creator>Neparidze, N.</dc:creator>
<dc:creator>Wong, E.</dc:creator>
<dc:creator>Torres, R.</dc:creator>
<dc:creator>Bruscia, E. M.</dc:creator>
<dc:creator>Kluger, Y.</dc:creator>
<dc:creator>Manz, M. G.</dc:creator>
<dc:creator>Flavell, R. A.</dc:creator>
<dc:creator>Halene, S.</dc:creator>
<dc:date>2018-02-14</dc:date>
<dc:identifier>doi:10.1101/265082</dc:identifier>
<dc:title><![CDATA[A Highly Efficient and Faithful MDS Patient-Derived Xenotransplantation Model for Pre-Clinical Studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/285940v1?rss=1">
<title>
<![CDATA[
Hierarchical modeling of the effect of pre-exposure prophylaxis on HIV in the US 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/285940v1?rss=1</link>
<description><![CDATA[
1.AO_SCPLOWBSTRACTC_SCPLOWIn this paper we present a differential equation model stratified by behavioral risk and sexual activity. Some susceptible individuals have higher rates of risky behavior that increase their chance of contracting the disease. Infected individuals can be considered to be generally sexually active or inactive. The sexually active infected population is at higher risk of transmitting the disease to a susceptible individual. We further divide the sexually active population into diagnosed or undiagnosed infected individuals. We define model parameters for both the national and the urban case. These parameter sets are used to study the predicted population dynamics over the next 5 years. Our results indicate that the undiagnosed high risk infected group is the largest contributor to the epidemic. Finally, we apply a preventative medication protocol to the susceptible population and observe the effective reduction in the infected population. The simulations suggest that preventative medication effectiveness extends outside of the group that is taking the drug (herd immunity). Our models suggest that a strategy targeting the high risk undiagnosed infected group would have the largest impact in the next 5 years. We also find that such a protocol has similar effects for the national as the urban case, despite the smaller sexual network found in rural areas.
]]></description>
<dc:creator>Dale, R.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>He, H.</dc:creator>
<dc:date>2018-03-22</dc:date>
<dc:identifier>doi:10.1101/285940</dc:identifier>
<dc:title><![CDATA[Hierarchical modeling of the effect of pre-exposure prophylaxis on HIV in the US]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/341313v1?rss=1">
<title>
<![CDATA[
Comprehensive 2D and 3D phenotypic characterization of human invasive lobular carcinoma cell lines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/341313v1?rss=1</link>
<description><![CDATA[
Invasive lobular carcinoma (ILC) is the second most common subtype of breast cancer following invasive ductal carcinoma (IDC) and characterized by the loss of E-cadherin-mediated adherens junctions. Despite displaying unique histological and clinical features, ILC still remains a chronically understudied disease with limited knowledge on the available laboratory research models. To this end, herein we report a comprehensive 2D and 3D phenotypic characterization of four Estrogen Receptor-positive human ILC cell lines - MDA-MB-134, SUM44, MDA-MB-330 and BCK4. Compared to the IDC cell lines MCF7, T47D and MDA-MB-231, ultra-low attachment culture conditions revealed a remarkable anchorage-independence ability that was unique to the ILC cells, a feature not evident in soft agar gels. 3D Collagen I and Matrigel culture indicated a generally loose morphology for the ILC cell lines, which exhibited differing preferences for adhesion to ECM proteins in 2D. Furthermore, ILC cells had limited migration and invasion ability in wound-scratch and transwell assays with the exception of haptotaxis to Collagen I. Transcriptional comparison of the cell lines confirmed the decreased cell proliferation and E-cadherin-mediated intercellular junctions in ILC, while uncovering the induction of novel pathways related to cyclic nucleotide phosphodiesterase activity, ion channels, drug metabolism and alternative cell adhesion molecules such as N-cadherin, some of which were also differentially regulated in ILC versus IDC tumors. Altogether, these studies will serve as an invaluable resource for the breast cancer research community and facilitate further functional discoveries towards understanding ILC, identifying novel drug targets and ultimately improving the outcome of patients with ILC.nnAuthors ContributionsConception and design: N. Tasdemir, NE. Davidson, S. OesterreichnnDevelopment of methodology: N. Tasdemir, L. Zhu, GC. Tseng, S. OesterreichnnAcquisition of data (performed experiments, processed data, etc.): N. Tasdemir, E. Bossart, Z. Li, Z. LinnAnalysis and interpretation of data (e.g. biological interpretation, statistical analysis, computational analysis): N. Tasdemir, Z. Li, KM. Levine, NE. Davidson, S. OesterreichnnWriting, review and/or revision of the manuscript: N. Tasdemir, Z. Li, KM. Levine, BM. Jacobson, GC. Tseng, NE. Davidson, S. OesterreichnnStudy supervision: NE. Davidson and S. Oesterreich
]]></description>
<dc:creator>Tasdemir, N.</dc:creator>
<dc:creator>Bossart, E.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Levine, K. M.</dc:creator>
<dc:creator>Jacobsen, B. M.</dc:creator>
<dc:creator>Tseng, G. C.</dc:creator>
<dc:creator>Davidson, N. E.</dc:creator>
<dc:creator>Oesterreich, S.</dc:creator>
<dc:date>2018-06-07</dc:date>
<dc:identifier>doi:10.1101/341313</dc:identifier>
<dc:title><![CDATA[Comprehensive 2D and 3D phenotypic characterization of human invasive lobular carcinoma cell lines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/371492v1?rss=1">
<title>
<![CDATA[
The splicing factor U2AF1 contributes to cancer progression through a non-canonical role in translation regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/371492v1?rss=1</link>
<description><![CDATA[
Somatic mutations in the genes encoding components of the spliceosome occur frequently in human neoplasms, including myeloid dysplasias and leukemias and less often in solid tumors. One of the affected factors, U2AF1, is involved in splice site selection, and the most common change, S34F, alters a conserved nucleic acid binding domain, recognition of the 3-splice site, and alternative splicing of many mRNAs. However, the role that this mutation plays in oncogenesis is still unknown. Here, we have uncovered a non-canonical function of U2AF1, showing that it binds mature mRNA in the cytoplasm and negatively regulates mRNA translation. This splicing-independent role of U2AF1 is altered by the S34F mutation, and polysomal profiling indicates the mutation affects translation of hundreds of mRNA. One functional consequence is increased synthesis of the secreted chemokine interleukin 8 which contributes to metastasis, inflammation, and cancer progression in mice and humans.
]]></description>
<dc:creator>Palangat, M.</dc:creator>
<dc:creator>Anastsakis, D.</dc:creator>
<dc:creator>Liang, F.</dc:creator>
<dc:creator>Lindblad, K.</dc:creator>
<dc:creator>Bradley, R.</dc:creator>
<dc:creator>Hourigan, C.</dc:creator>
<dc:creator>Hafner, M.</dc:creator>
<dc:creator>Larson, D.</dc:creator>
<dc:date>2018-07-18</dc:date>
<dc:identifier>doi:10.1101/371492</dc:identifier>
<dc:title><![CDATA[The splicing factor U2AF1 contributes to cancer progression through a non-canonical role in translation regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/675272v1?rss=1">
<title>
<![CDATA[
Identification of antibodies targeting the H3N2 hemagglutinin receptor binding site following vaccination of humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/675272v1?rss=1</link>
<description><![CDATA[
Antibodies targeting the receptor binding site (RBS) of the influenza virus hemagglutinin (HA) protein are usually not broadly-reactive because their footprints are typically large and extend to nearby variable HA residues. Here, we identified several human H3N2 HA RBS-targeting monoclonal antibodies (mAbs) that were sensitive to substitutions in conventional antigenic sites and were not broadly-reactive. However, we also identified one H3N2 HA RBS-targeting mAb that was exceptionally broadly reactive despite being sensitive to substitutions in residues outside of the RBS. We determined that similar antibodies are present at measurable levels in the sera of some individuals but that they are inefficiently elicited by conventional vaccines. Our data indicate that some HA RBS-targeting antibodies can be surprisingly effective against variable viral strains even if they are somewhat sensitive to substitutions in HA residues adjacent to the RBS.
]]></description>
<dc:creator>Zost, S. J.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Gumina, M. E.</dc:creator>
<dc:creator>Parkhouse, K.</dc:creator>
<dc:creator>Henry, C.</dc:creator>
<dc:creator>Wilson, P. C.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Hensley, S. E.</dc:creator>
<dc:date>2019-06-19</dc:date>
<dc:identifier>doi:10.1101/675272</dc:identifier>
<dc:title><![CDATA[Identification of antibodies targeting the H3N2 hemagglutinin receptor binding site following vaccination of humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/013623v1?rss=1">
<title>
<![CDATA[
Software for the analysis and visualization of deep mutational scanning data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/013623v1?rss=1</link>
<description><![CDATA[
BackgroundDeep mutational scanning is a technique to estimate the impacts of mutations on a gene by using deep sequencing to count mutations in a library of variants before and after imposing a functional selection. The impacts of mutations must be inferred from changes in their counts after selection.nnResultsI describe a software package, dms_tools, to infer the impacts of mutations from deep mutational scanning data using a likelihood-based treatment of the mutation counts. I show that dms_tools yields more accurate inferences on simulated data than simply calculating ratios of counts pre-and post-selection. Using dms_tools, one can infer the preference of each site for each amino acid given a single selection pressure, or assess the extent to which these preferences change under different selection pressures. The preferences and their changes can be intuitively visualized with sequence-logo-style plots created using an extension to weblogo.nnConclusionsdms_tools implements a statistically principled approach for the analysis and subsequent visualization of deep mutational scanning data.
]]></description>
<dc:creator>Jesse D Bloom</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-01-10</dc:date>
<dc:identifier>doi:10.1101/013623</dc:identifier>
<dc:title><![CDATA[Software for the analysis and visualization of deep mutational scanning data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/026567v1?rss=1">
<title>
<![CDATA[
Immunosequencing reveals diagnostic signatures of chronic viral infection in T cell memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/026567v1?rss=1</link>
<description><![CDATA[
B and T cells expand clonally in response to pathogenic infection, and their descendants, which share the same receptor sequence, can persist for years, forming the basis of immunological memory. While most T cell receptor (TCR) sequences are seen very rarely,  public TCRs are present in many individuals.nnUsing a combination of high throughput immunosequencing, statistical association of particular TCRs with disease status, and machine learning, we identified of a set of public TCRs that discriminates cytomegalovirus (CMV) infection status with high accuracy. This pathogen-specific diagnostic tool uses a very general assay that relies only on a training cohort coupled with immunosequencing and sophisticated data analysis. Since all memory T cell responses are encoded in the common format of somatic TCR rearrangements, a key advantage of reading T cell memory to predict disease status is that this approach should apply to a wide variety of diseases. The underlying dataset is the largest collection of TCRs ever published, including [~]300 gigabases of sequencing data and [~]85 million unique TCRs across 640 HLA-typed individuals, which constitutes by far the largest such collection ever published. We expect these data to be a valuable public resource for researchers studying the TCR repertoire.
]]></description>
<dc:creator>Ryan Emerson</dc:creator>
<dc:creator>William DeWitt</dc:creator>
<dc:creator>Marissa Vignali</dc:creator>
<dc:creator>Jenna Gravley</dc:creator>
<dc:creator>Cindy Desmarais</dc:creator>
<dc:creator>Christoper Carlson</dc:creator>
<dc:creator>John Hansen</dc:creator>
<dc:creator>Mark Rieder</dc:creator>
<dc:creator>Harlan Robins</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-09-10</dc:date>
<dc:identifier>doi:10.1101/026567</dc:identifier>
<dc:title><![CDATA[Immunosequencing reveals diagnostic signatures of chronic viral infection in T cell memory]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/032300v1?rss=1">
<title>
<![CDATA[
Conjecture: Some auto-immune diseases are caused by the tissue, not a problem with the immune system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/032300v1?rss=1</link>
<description><![CDATA[
Applying highly sensitive modern immune profiling techniques to the blood or inflamed tissue of auto-immune patients with one of the common T-cell mediated diseases fails to detect large clonal expansions or other signs of a dysregulated immune system. Additionally, these new methods have shown that self-reactive T-cells are found in the periphery (and are not completely negatively selected in the thymus as previously believed). Combining these new data with well-established studies in auto-immunity leads to the conjecture that certain auto-immune diseases are likely to be caused by defects in tissue, not by dysregulation of the adaptive immune system. In particular, one hypothesis is that specific tissues up-regulate HLA class 2 genes, presenting epitopes that are bound by CD4+ helper T cells, which facilitates an immune response against the tissue specific cells.
]]></description>
<dc:creator>Harlan S Robins</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-11-20</dc:date>
<dc:identifier>doi:10.1101/032300</dc:identifier>
<dc:title><![CDATA[Conjecture: Some auto-immune diseases are caused by the tissue, not a problem with the immune system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/034082v1?rss=1">
<title>
<![CDATA[
Winners curse correction and variable thresholding improve performance of polygenic risk modeling based on summary-level data from genome-wide association studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/034082v1?rss=1</link>
<description><![CDATA[
Recent heritability analyses have indicated that genome-wide association studies (GWAS) have the potential to improve genetic risk prediction for complex diseases based on polygenic risk score (PRS), a simple modelling technique that can be implemented using summary-level data from the discovery samples. We herein propose modifications to improve the performance of PRS. We introduce threshold-dependent winners-curse adjustments for marginal association coefficients that are used to weight the SNPs in PRS. Further, as a way to incorporate external functional/annotation knowledge that could identify subsets of SNPs highly enriched for associations, we propose variable thresholds for SNPs selection. We applied our methods to GWAS summary-level data of 14 complex diseases. Across all diseases, a simple winners curse correction uniformly led to enhancement of performance of the models, whereas incorporation of functional SNPs was beneficial only for selected diseases. Compared to the standard PRS algorithm, the proposed methods in combination led to notable gain in efficiency (25-50% increase in the prediction R2) for 5 of 14 diseases. As an example, for GWAS of type 2 diabetes, winners curse correction improved prediction R2 from 2.29% based on the standard PRS to 3.10% (P=0.0017) and incorporating functional annotation data further improved R2 to 3.53% (P=2{chi}10-5). Our simulation studies illustrate why differential treatment of certain categories of functional SNPs, even when shown to be highly enriched for GWAS-heritability, does not lead to proportionate improvement in genetic risk-prediction because of non-uniform linkage disequilibrium structure.
]]></description>
<dc:creator>Jianxin Shi</dc:creator>
<dc:creator>JuHyun Park</dc:creator>
<dc:creator>Jubao Duan</dc:creator>
<dc:creator>Sonja Berndt</dc:creator>
<dc:creator>Winton Moy</dc:creator>
<dc:creator>William Wheeler</dc:creator>
<dc:creator>Xing Hua</dc:creator>
<dc:creator>Debra Silverman</dc:creator>
<dc:creator>Montserrat Garcia-Closas</dc:creator>
<dc:creator>Chao Agnes Hsiung</dc:creator>
<dc:creator>Jonine D. Figueroa</dc:creator>
<dc:creator>Victoria K. Cortessis</dc:creator>
<dc:creator>Nuria Malats</dc:creator>
<dc:creator>Margaret R. Karagas</dc:creator>
<dc:creator>Paolo Vineis</dc:creator>
<dc:creator>I-Shou Chang</dc:creator>
<dc:creator>Dongxin Lin</dc:creator>
<dc:creator>Baosen Zhou</dc:creator>
<dc:creator>Adeline Seow</dc:creator>
<dc:creator>Keitaro Matsuo</dc:creator>
<dc:creator>Yun-Chul Hong</dc:creator>
<dc:creator>Neil E. Caporaso</dc:creator>
<dc:creator>Brian Wolpin</dc:creator>
<dc:creator>Eric Jacobs</dc:creator>
<dc:creator>Gloria Petersen</dc:creator>
<dc:creator>Donghui Li</dc:creator>
<dc:creator>Harvey Risch</dc:creator>
<dc:creator>Alan R. Sanders</dc:creator>
<dc:creator>Li Hsu</dc:creator>
<dc:creator>Robert E. Schoen</dc:creator>
<dc:creator>Hermann Brenner</dc:creator>
<dc:creator>Rachael Stolzenberg-Solomon</dc:creator>
<dc:creator>Pablo Gejman</dc:creator>
<dc:creator>Qing Lan</dc:creator>
<dc:creator>Nathaniel Rothman</dc:creator>
<dc:creator>Laufey T. Amundadottir</dc:creator>
<dc:creator>Alison P. Klein</dc:creator>
<dc:creator>Maria Teresa Landi</dc:creator>
<dc:creator>Douglas F. Levinson</dc:creator>
<dc:creator>Stephen</dc:creator>
<dc:date>2016-01-10</dc:date>
<dc:identifier>doi:10.1101/034082</dc:identifier>
<dc:title><![CDATA[Winners curse correction and variable thresholding improve performance of polygenic risk modeling based on summary-level data from genome-wide association studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/038687v1?rss=1">
<title>
<![CDATA[
Translational plasticity facilitates the accumulation of nonsense genetic variants in the human population 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/038687v1?rss=1</link>
<description><![CDATA[
Genetic variants that disrupt protein-coding DNA are ubiquitous in the human population, with ~100 such loss-of-function variants per individual. While most loss-of-function variants are rare, a subset have risen to high frequency and occur in a homozygous state in healthy individuals. It is unknown why these common variants are well-tolerated, even though some affect essential genes implicated in Mendelian disease. Here, we combine genomic, proteomic, and biochemical data to demonstrate that many common nonsense variants do not ablate protein production from their host genes. We provide computational and experimental evidence for diverse mechanisms of gene rescue, including alternative splicing, stop codon readthrough, alternative translation initiation, and C-terminal truncation. Our results suggest a molecular explanation for the mild fitness costs of many common nonsense variants, and indicate that translational plasticity plays a prominent role in shaping human genetic diversity.
]]></description>
<dc:creator>Sujatha Jagannathan</dc:creator>
<dc:creator>Robert K. Bradley</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-02-04</dc:date>
<dc:identifier>doi:10.1101/038687</dc:identifier>
<dc:title><![CDATA[Translational plasticity facilitates the accumulation of nonsense genetic variants in the human population]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/039578v1?rss=1">
<title>
<![CDATA[
Cryopreservation of human mucosal leukocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/039578v1?rss=1</link>
<description><![CDATA[
BackgroundUnderstanding how leukocytes in the cervicovaginal and colorectal mucosae respond to pathogens, and how medical interventions affect these responses, is important for developing better tools to prevent HIV and other sexually transmitted infections. An effective cryopreservation protocol for these cells following their isolation will make studying them more feasible.nnMethods and findingsTo find an optimal cryopreservation protocol for mucosal mononuclear leukocytes, we compared cryopreservation media and procedures using human vaginal leukocytes and confirmed our results with endocervical and colorectal leukocytes. Specifically, we measured the recovery of viable vaginal T cells and macrophages after cryopreservation with different cryopreservation media and handling procedures. We found several cryopreservation media that led to recoveries above 75%. Limiting the number and volume of washes increased the fraction of cells recovered by 10-15%, possibly due to the small cell numbers in mucosal samples. We confirmed that our cryopreservation protocol also works well for both endocervical and colorectal leukocytes.nnCryopreserved leukocytes had slightly increased cytokine responses to antigenic stimulation relative to the same cells tested fresh. Additionally, we tested whether it is better to cryopreserve endocervical cells on the cytobrush or in suspension.nnConclusionsLeukocytes from cervicovaginal and colorectal tissues can be cryopreserved with good recovery of functional, viable cells using several different cryopreservation media. The number and volume of washes has an experimentally meaningful effect on the percentage of cells recovered. We provide a detailed, step-by-step protocol with best practices for cryopreservation of mucosal leukocytes.
]]></description>
<dc:creator>Sean M. Hughes</dc:creator>
<dc:creator>Zhiquan Shu</dc:creator>
<dc:creator>Claire N. Levy</dc:creator>
<dc:creator>April L. Ferre</dc:creator>
<dc:creator>Heather Hartig</dc:creator>
<dc:creator>Cifeng Fang</dc:creator>
<dc:creator>Gretchen Lentz</dc:creator>
<dc:creator>Michael Fialkow</dc:creator>
<dc:creator>Anna C Kirby</dc:creator>
<dc:creator>Kristina M Adams Waldorf</dc:creator>
<dc:creator>Ronald S Veazey</dc:creator>
<dc:creator>Anja Germann</dc:creator>
<dc:creator>Hagen von Briesen</dc:creator>
<dc:creator>M. Juliana McElrath</dc:creator>
<dc:creator>Charlene S Dezzutti</dc:creator>
<dc:creator>Elizabeth Sinclair</dc:creator>
<dc:creator>Chris A. R. Baker</dc:creator>
<dc:creator>Barbara L Shacklett</dc:creator>
<dc:creator>Dayong Gao</dc:creator>
<dc:creator>Florian Hladik</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-03-24</dc:date>
<dc:identifier>doi:10.1101/039578</dc:identifier>
<dc:title><![CDATA[Cryopreservation of human mucosal leukocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/048215v1?rss=1">
<title>
<![CDATA[
Extracting a low-dimensional description of multiple gene expression datasets reveals a potential driver for tumor-associated stroma in ovarian cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/048215v1?rss=1</link>
<description><![CDATA[
BackgroundDiscovering patient subtypes and molecular drivers of a subtype are difficult and driving problems underlying most modern disease expression studies collected across patient populations. Expression patterns conserved across multiple expression datasets from independent disease studies are likely to represent important molecular events underlying the disease.nnMethodsWe present the INSPIRE (INferring Shared modules from multiPle gene expREssion datasets) method to infer highly coherent and robust modules of co-expressed genes and the dependencies among the modules from multiple expression datasets. Focusing on inferring modules and their dependencies conserved across multiple expression datasets is important for several reasons. First, using multiple datasets will increase the power to detect robust and relevant patterns (modules and dependencies among modules). Second, INSPIRE enables the use of multiple datasets that contain different sets of genes due to, e.g., the difference in microarray platforms. Many methods designed for expression data analysis cannot integrate multiple datasets with variable discrepancy to infer a single combined model, whereas INSPIRE can naturally model the dependencies among the modules even when a large proportion of genes are not observed on a certain platform.nnResultsWe evaluated INSPIRE on synthetically generated datasets with known underlying network structure among modules, and gene expression datasets from multiple ovarian cancer studies. We show that the model learned by INSPIRE can explain unseen data better and can reveal prior knowledge on gene functions more accurately than alternative methods. We demonstrate that applying INSPIRE to nine ovarian cancer datasets leads to the identification of a new marker and potential molecular driver of tumor-associated stroma - HOPX. We also demonstrate that the HOPXmodule strongly overlaps with the genes defining the mesenchymal patient subtype identified in The Cancer Genome Atlas (TCGA) ovarian cancer data. We provide evidence for a previously unknown molecular basis of tumor resectability efficacy involving tumor-associated mesenchymal stem cells represented by HOPX.nnConclusionsINSPIRE extracts a low-dimensional description from multiple gene expression data, which consists of modules and their dependencies. The discovery of a new tumor-associated stroma marker, HOPX, and its module suggests a previously unknown mechanism underlying tumor-associated stroma.
]]></description>
<dc:creator>Safiye Celik</dc:creator>
<dc:creator>Benjamin A Logsdon</dc:creator>
<dc:creator>Stephanie Battle</dc:creator>
<dc:creator>Charles W Drescher</dc:creator>
<dc:creator>Mara Rendi</dc:creator>
<dc:creator>David Hawkins</dc:creator>
<dc:creator>Su-In Lee</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-04-13</dc:date>
<dc:identifier>doi:10.1101/048215</dc:identifier>
<dc:title><![CDATA[Extracting a low-dimensional description of multiple gene expression datasets reveals a potential driver for tumor-associated stroma in ovarian cancer]]></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/10.1101/060988v1?rss=1">
<title>
<![CDATA[
The validity of pairwise models in predicting community dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/060988v1?rss=1</link>
<description><![CDATA[
Pairwise models are commonly used to describe many-species communities. In these models, a focal species receives additive fitness effects from pairwise interactions with other species in the community ("pairwise additivity assumption"), and all pairwise interactions are represented by a single canonical equation form ("universality assumption"). Here, we analyze the validity of pairwise modeling. We build mechanistic reference models for chemical-mediated interactions in microbial communities, and attempt to derive corresponding pairwise models. Even when one species affects another via a single chemical mediator, different forms of pairwise models are appropriate for consumable versus reusable mediators, with the wrong model producing qualitatively wrong predictions. For multi-mediator interactions, a canonical model becomes even less tenable. These results, combined with potential violation of the pairwise additivity assumption in communities of more than two species, suggest that although pairwise modeling can be useful, we should examine its validity before employing it.
]]></description>
<dc:creator>Babak Momeni</dc:creator>
<dc:creator>Li Xie</dc:creator>
<dc:creator>Wenying Shou</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-06-27</dc:date>
<dc:identifier>doi:10.1101/060988</dc:identifier>
<dc:title><![CDATA[The validity of pairwise models in predicting community dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/063305v1?rss=1">
<title>
<![CDATA[
A compound interest approach to HIV cure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/063305v1?rss=1</link>
<description><![CDATA[
In the era of antiretroviral therapy (ART), HIV-1 infection is no longer tantamount to early death. Yet the benefits of treatment are available only to those who can access, afford, and tolerate taking daily pills. True cure is challenged by HIV latency, the ability of chromosomally integrated virus to persist within memory CD4+ T cells in a non-replicative state and activate when ART is discontinued. Using a mathematical model of HIV dynamics, we demonstrate that treatment strategies offering modest but continual enhancement of reservoir clearance rates result in faster cure than abrupt, one-time reductions in reservoir size. We frame this concept in terms of compounding interest: small changes in interest rate drastically improve returns over time. On ART, latent cell proliferation rates are orders of magnitude larger than activation and new infection rates. Contingent on subtypes of cells that may make up the reservoir and their respective proliferation rates, our model predicts that coupling clinically available, anti-proliferative therapies with ART could result in functional cure within 2-10 years rather than several decades on ART alone.
]]></description>
<dc:creator>Daniel Reeves</dc:creator>
<dc:creator>Elizabeth Duke</dc:creator>
<dc:creator>Martin Prlic</dc:creator>
<dc:creator>Florian Hladik</dc:creator>
<dc:creator>Joshua Schiffer</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-07-12</dc:date>
<dc:identifier>doi:10.1101/063305</dc:identifier>
<dc:title><![CDATA[A compound interest approach to HIV cure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/065912v1?rss=1">
<title>
<![CDATA[
Massively parallel digital transcriptional profiling of single cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/065912v1?rss=1</link>
<description><![CDATA[
Characterizing the transcriptome of individual cells is fundamental to understanding complex biological systems. We describe a droplet-based system that enables 3' mRNA counting of up to tens of thousands of single cells per sample. Cell encapsulation in droplets takes place in [~]6 minutes, with [~]50% cell capture efficiency, up to 8 samples at a time. The speed and efficiency allow the processing of precious samples while minimizing stress to cells. To demonstrate the system's technical performance and its applications, we collected transcriptome data from [~][1/4] million single cells across 29 samples. First, we validate the sensitivity of the system and its ability to detect rare populations using cell lines and synthetic RNAs. Then, we profile 68k peripheral blood mononuclear cells (PBMCs) to demonstrate the system's ability to characterize large immune populations. Finally, we use sequence variation in the transcriptome data to determine host and donor chimerism at single cell resolution in bone marrow mononuclear cells (BMMCs) of transplant patients. This analysis enables characterization of the complex interplay between donor and host cells and monitoring of treatment response. This high-throughput system is robust and enables characterization of diverse biological systems with single cell mRNA analysis.
]]></description>
<dc:creator>Grace X.Y. Zheng</dc:creator>
<dc:creator>Jessica M Terry</dc:creator>
<dc:creator>Phillip Belgrader</dc:creator>
<dc:creator>Paul Ryvkin</dc:creator>
<dc:creator>Zachary W. Bent</dc:creator>
<dc:creator>Ryan Wilson</dc:creator>
<dc:creator>Solongo B. Ziraldo</dc:creator>
<dc:creator>Tobias D. Wheeler</dc:creator>
<dc:creator>Geoff P. McDermott</dc:creator>
<dc:creator>Junjie Zhu</dc:creator>
<dc:creator>Mark T. Gregory</dc:creator>
<dc:creator>Joe Shuga</dc:creator>
<dc:creator>Luz Montesclaros</dc:creator>
<dc:creator>Donald A Masquelier</dc:creator>
<dc:creator>Stefanie Y. Nishimura</dc:creator>
<dc:creator>Michael Schnall-Levin</dc:creator>
<dc:creator>Paul W Wyatt</dc:creator>
<dc:creator>Christopher M. Hindson</dc:creator>
<dc:creator>Rajiv Bharadwaj</dc:creator>
<dc:creator>Alexander Wong</dc:creator>
<dc:creator>Kevin D. Ness</dc:creator>
<dc:creator>Lan W. Beppu</dc:creator>
<dc:creator>Joachim Deeg</dc:creator>
<dc:creator>Christopher McFarland</dc:creator>
<dc:creator>Keith R. Loeb</dc:creator>
<dc:creator>William J. Valente</dc:creator>
<dc:creator>Nolan G. Ericson</dc:creator>
<dc:creator>Emily A. Stevens</dc:creator>
<dc:creator>Jerald P. Radich</dc:creator>
<dc:creator>Tarjei S. Mikkelsen</dc:creator>
<dc:creator>Benjamin J. Hindson</dc:creator>
<dc:creator>Jason H Bielas</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-07-26</dc:date>
<dc:identifier>doi:10.1101/065912</dc:identifier>
<dc:title><![CDATA[Massively parallel digital transcriptional profiling of single cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/067470v1?rss=1">
<title>
<![CDATA[
Experimental estimation of the effects of all amino-acid mutations to HIV Env 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/067470v1?rss=1</link>
<description><![CDATA[
HIV is notorious for its capacity to evade immunity and anti-viral drugs through rapid sequence evolution. Knowledge of the functional effects of mutations to HIV is critical for understanding this evolution. HIVs most rapidly evolving protein is its envelope (Env). Here we use deep mutational scanning to experimentally estimate the effects of all amino-acid mutations to Env on viral replication in cell culture. Most mutations are under purifying selection in our experiments, although a few sites experience strong selection for mutations that enhance HIVs growth in cell culture. We compare our experimental measurements of each sites preference for each amino acid to the actual frequencies of these amino acids in naturally occurring HIV sequences. Our measured amino-acid preferences correlate with amino-acid frequencies in natural sequences for most sites. However, our measured preferences are less concordant with natural amino-acid frequencies at surface-exposed sites that are subject to pressures absent from our experiments such as antibody selection. We show that some regions of Env have a high inherent tolerance to mutation, whereas other regions (such as epitopes of broadly neutralizing antibodies) have a significantly reduced capacity to tolerate mutations. Overall, our results help disentangle the role of inherent functional constraints and external selection pressures in shaping Envs evolution.
]]></description>
<dc:creator>Hugh K Haddox</dc:creator>
<dc:creator>Adam S Dingens</dc:creator>
<dc:creator>Jesse Bloom</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-08-02</dc:date>
<dc:identifier>doi:10.1101/067470</dc:identifier>
<dc:title><![CDATA[Experimental estimation of the effects of all amino-acid mutations to HIV Env]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/071969v1?rss=1">
<title>
<![CDATA[
Deep mutational scanning identifies sites in influenza nucleoprotein that affect viral inhibition by MxA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/071969v1?rss=1</link>
<description><![CDATA[
The innate-immune restriction factor MxA inhibits influenza replication by targeting the viral nucleoprotein (NP). Human influenza is more resistant than avian influenza to inhibition by human MxA, and prior work has compared human and avian viral strains to identify amino-acid differences in NP that affect sensitivity to MxA. However, this strategy is limited to identifying sites in NP where mutations that affect MxA sensitivity have fixed during the small number of documented zoonotic transmissions of influenza to humans. Here we use an unbiased deep mutational scanning approach to quantify how all {approx}10,000 amino-acid mutations to NP affect MxA sensitivity. We both identify new sites in NP where mutations affect MxA resistance and re-identify mutations known to have increased MxA resistance during historical adaptations of influenza to humans. Most of the sites where mutations have the greatest effect are almost completely conserved across all influenza A viruses, and the amino acids at these sites confer relatively high resistance to MxA. These sites cluster in regions of NP that appear to be important for its recognition by MxA. Overall, our work systematically identifies the sites in influenza nucleoprotein where mutations affect sensitivity to MxA. We also demonstrate a powerful new strategy for identifying regions of viral proteins that affect interactions with host factors.nnAuthor SummaryDuring viral infection, human cells express proteins that can restrict virus replication. However, in many cases it remains unclear what determines the sensitivity of a given viral strain to a particular restriction factor. Here we use a high-throughput approach to measure how all amino-acid mutations to the nucleoprotein of influenza virus affect restriction by the human protein MxA. We find several dozen sites where mutations substantially affect influenzas sensitivity to MxA. While a few of these sites are known to have fixed mutations during past adaptations of influenza to humans, most of the sites are broadly conserved across all influenza strains and have never previously been described as affecting MxA resistance. Our results therefore show that the known historical evolution of influenza has only involved substitutions at a small fraction of the sites where mutations can in principle affect MxA resistance. We suggest that this is because many sites are already broadly fixed at amino acids that confer high resistance.
]]></description>
<dc:creator>Orr Ashenberg</dc:creator>
<dc:creator>Jai Padmakumar</dc:creator>
<dc:creator>Michael B Doud</dc:creator>
<dc:creator>Jesse Bloom</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-08-28</dc:date>
<dc:identifier>doi:10.1101/071969</dc:identifier>
<dc:title><![CDATA[Deep mutational scanning identifies sites in influenza nucleoprotein that affect viral inhibition by MxA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/076638v1?rss=1">
<title>
<![CDATA[
Seasonal influenza circulation patterns and projections for 2016-2017 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/076638v1?rss=1</link>
<description><![CDATA[
This is not meant as a comprehensive report, but is instead intended as particular observations that may be of relevance. Please also note that observed patterns reflect the GISAID database and may not be entirely representative of underlying dynamics. All analyses are based on the nextflu pipeline [1] with continual updates posted to Correspondence:http://nextflu.org.
]]></description>
<dc:creator>Trevor Bedford</dc:creator>
<dc:creator>Richard Neher</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-09-21</dc:date>
<dc:identifier>doi:10.1101/076638</dc:identifier>
<dc:title><![CDATA[Seasonal influenza circulation patterns and projections for 2016-2017]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/086611v1?rss=1">
<title>
<![CDATA[
Complete mapping of viral escape from neutralizing antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/086611v1?rss=1</link>
<description><![CDATA[
Identifying viral mutations that confer escape from antibodies is crucial for understanding the interplay between immunity and viral evolution. Here we quantify how every amino-acid mutation to influenza hemagglutinin affects neutralization by monoclonal antibodies targeting several antigenic regions. Our approach involves creating all replication-competent protein variants of the virus, selecting these variants with antibody, and using deep sequencing to identify enriched mutations. These high-throughput measurements are predictive of the effects of individual mutations in traditional neutralization assays. At many residues, only some of the possible mutations escape from an antibody. For instance, at a single residue targeted by two different antibodies, we identify some mutations that escape both antibodies and other mutations that escape only one or the other. Therefore, our approach maps how viruses can escape antibodies with mutation-level sensitivity, and shows that only some mutations at antigenic residues actually alter antigenicity.
]]></description>
<dc:creator>Doud, M. B.</dc:creator>
<dc:creator>Hensley, S. E.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2016-11-09</dc:date>
<dc:identifier>doi:10.1101/086611</dc:identifier>
<dc:title><![CDATA[Complete mapping of viral escape from neutralizing antibodies]]></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/10.1101/101725v1?rss=1">
<title>
<![CDATA[
Modern-Day SIV viral diversity generated by extensive recombination and cross-species transmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/101725v1?rss=1</link>
<description><![CDATA[
Cross-species transmission (CST) has led to many devastating epidemics, but is still a poorly understood phenomenon. HIV-1 and HIV-2 (human immunodeficiency virus 1 and 2), which have collectively caused over 35 million deaths, are the result of multiple CSTs from chimpanzees, gorillas, and sooty mangabeys. While the immediate history of HIV is known, there are over 45 lentiviruses that infect specific species of primates, and patterns of host switching are not well characterized. We thus took a phylogenetic approach to better understand the natural history of SIV recombination and CST. We modeled host species as a discrete character trait on the viral phylogeny and inferred historical host switches and the pairwise transmission rates between each pair of 24 primate hosts. We identify 14 novel, well-supported, ancient cross-species transmission events. We also find that lentiviral lineages vary widely in their ability to infect new host species: SIVcol (from colobus monkeys) is evolutionarily isolated, while SIVagms (from African green monkeys) frequently move between host subspecies. We also examine the origins of SIVcpz (the predecessor of HIV-1) in greater detail than previous studies, and find that there are still large portions of the genome with unknown origins. Observed patterns of CST are likely driven by a combination of ecological circumstance and innate immune factors.
]]></description>
<dc:creator>Bell, S. M.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:date>2017-01-19</dc:date>
<dc:identifier>doi:10.1101/101725</dc:identifier>
<dc:title><![CDATA[Modern-Day SIV viral diversity generated by extensive recombination and cross-species transmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/113035v1?rss=1">
<title>
<![CDATA[
Seasonal influenza circulation patterns and projections for 2017-2018 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/113035v1?rss=1</link>
<description><![CDATA[
This is not meant as a comprehensive report of recent influenza evolution, but is instead intended as particular observations that may be of relevance. Please also note that observed patterns reflect the GISAID database and may not be entirely representative of underlying dynamics. All analyses are based on the nextflu pipeline [1] with continual updates posted to nextflu.org. We arrive at the following results:nnH3N2In H3N2, clade 3c2.a has continued to diversify genetically with complicated and rapid dynamics of different subclades. This diversification is not reflected in serological data that shows only minor to moderate antigenic evolution. Nevertheless, the highly parallel mutation patterns and the rapid rise and fall of clades suggests competitive dynamics of phenotypically distinct viruses.nnH1N1pdmVery few H1N1pdm viruses have been observed in recent months. The dominant clade continues to be 6b.1 and there is little amino acid sequence variation within HA. The only notable subclade that has been growing recently is the clade bearing HA1:R205K/S183P. This clade is dominated by North American viruses and we see no evidence that this clade has a particular competitive advantage.nnB/VicClade 1A has continued to dominate and mutation 117V has all but taken over the global population. The rise of this mutation was fairly gradual and we have no evidence that it is associated with antigenic change or other benefit to the virus.nnB/YamClade 3 has continued to dominate. Within clade 3, a clade with mutation HA1:251V is globally at frequency of about 80% throughout 2016. Within this clade, mutation 211R is at 25% frequency. In addition, a clade without prominent amino acid mutations has been rising throughout 2016.
]]></description>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:date>2017-03-02</dc:date>
<dc:identifier>doi:10.1101/113035</dc:identifier>
<dc:title><![CDATA[Seasonal influenza circulation patterns and projections for 2017-2018]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/113100v1?rss=1">
<title>
<![CDATA[
Genetic characterization of the Zika virus epidemic in the US Virgin Islands 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/113100v1?rss=1</link>
<description><![CDATA[
Here we release draft genome sequences of Zika virus (ZIKV) that were sequenced from PCR-positive diagnostic specimens collected by the United States Virgin Islands Department of Health (USVI DoH) as part of their ongoing response to the ZIKV outbreak. We will use these sequences to conduct a genomic epidemiological study of ZIKV transmission in the USVI. We are releasing these genomes in the hope that they are useful for those individuals involved in the public health response to ZIKV and to other groups working to understand Zika virus transmission and evolution.
]]></description>
<dc:creator>Black, A.</dc:creator>
<dc:creator>Potter, B.</dc:creator>
<dc:creator>Dudas, G.</dc:creator>
<dc:creator>Feldstein, L.</dc:creator>
<dc:creator>Grubaugh, N. D.</dc:creator>
<dc:creator>Andersen, K. G.</dc:creator>
<dc:creator>Ellis, B. R.</dc:creator>
<dc:creator>Ellis, E. M.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:date>2017-03-03</dc:date>
<dc:identifier>doi:10.1101/113100</dc:identifier>
<dc:title><![CDATA[Genetic characterization of the Zika virus epidemic in the US Virgin Islands]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/117705v1?rss=1">
<title>
<![CDATA[
Comprehensive mapping of HIV-1 escape from a broadly neutralizing antibody 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/117705v1?rss=1</link>
<description><![CDATA[
Precisely defining how viral mutations affect HIVs sensitivity to antibodies is vital to the development and evaluation of vaccines and antibody immunotherapeutics. But despite great effort, a full map of escape mutants has not yet been delineated for even a single anti-HIV antibody. Here we describe a massively parallel experimental approach to quantify how all single amino-acid mutations to Envelope (Env) affect HIVs sensitivity to a neutralizing antibody. We applied this approach to PGT151 and identified novel sites of escape in addition to those previously defined by structural and functional studies, such as glycans at sites 611 and 637, residue 647, and sites in the fusion peptide. Evaluating the effect of each amino acid at each site lends insight into the biochemical basis of escape throughout the epitope. Thus, comprehensive mapping of HIV antibody escape gives a quantitative, mutation-level view of the ways that Env can evade neutralization.
]]></description>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2017-03-18</dc:date>
<dc:identifier>doi:10.1101/117705</dc:identifier>
<dc:title><![CDATA[Comprehensive mapping of HIV-1 escape from a broadly neutralizing antibody]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/117945v1?rss=1">
<title>
<![CDATA[
Brain connectivity-informed regularization methods for regression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/117945v1?rss=1</link>
<description><![CDATA[
A challenging problem arising in brain imaging research is principled incorporation of information from different imaging modalities. Frequently each modality is analyzed separately using, for instance, dimensionality reduction techniques which result in a loss of mutual information. We propose a novel regularization method to estimate the association between the brain structure features and a scalar outcome within the linear regression framework. Our regularization technique provides a principled approach to utilizing external information arising from the structural brain connectivity to inform the estimation of the regression coefficients. Our proposal extends the classical Tikhonov regularization framework by defining a penalty term based on the structural connectivity-derived Laplacian matrix. In the work presented, we address both theoretical and computational issues. The approach is illustrated using simulated data and compared with other penalized regression methods. Finally, we apply our regularization method to study the associations between the alcoholism phenotypes and brain cortical thickness using a diffusion tensor imaging (DTI) derived measure of structural connectivity.
]]></description>
<dc:creator>Karas, M.</dc:creator>
<dc:creator>Brzyski, D.</dc:creator>
<dc:creator>Dzemidzic, M.</dc:creator>
<dc:creator>Goni, J.</dc:creator>
<dc:creator>Kareken, D. A.</dc:creator>
<dc:creator>Randolph, T. W.</dc:creator>
<dc:creator>Harezlak, J.</dc:creator>
<dc:date>2017-03-18</dc:date>
<dc:identifier>doi:10.1101/117945</dc:identifier>
<dc:title><![CDATA[Brain connectivity-informed regularization methods for regression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/119545v1?rss=1">
<title>
<![CDATA[
Phandango: an interactive viewer for bacterial population genomics. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/119545v1?rss=1</link>
<description><![CDATA[
SummaryFully exploiting the wealth of data in current bacterial population genomics datasets requires synthesising and integrating different types of analysis across millions of base pairs in hundreds or thousands of isolates. Current approaches often use static representations of phylogenetic, epidemiological, statistical and evolutionary analysis results that are difficult to relate to one another. Phandango is an interactive application running in a web browser allowing fast exploration of large-scale population genomics datasets combining the output from multiple genomic analysis methods in an intuitive and interactive manner.nnAvailabilityPhandango is a web application freely available for use at https://jameshadfield.github.io/phandango and includes a diverse collection of datasets as examples. Source code together with a detailed wiki page is available on GitHub at https://github.com/jameshadfield/phandangonnContactjh22@sanger.ac.uk, sh16@sanger.ac.uk
]]></description>
<dc:creator>Hadfield, J.</dc:creator>
<dc:creator>Croucher, N. J.</dc:creator>
<dc:creator>Goater, R. J.</dc:creator>
<dc:creator>Abudahab, K.</dc:creator>
<dc:creator>Aanensen, D. M.</dc:creator>
<dc:creator>Harris, S. R.</dc:creator>
<dc:date>2017-03-22</dc:date>
<dc:identifier>doi:10.1101/119545</dc:identifier>
<dc:title><![CDATA[Phandango: an interactive viewer for bacterial population genomics.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/121830v1?rss=1">
<title>
<![CDATA[
phydms: Software for phylogenetic analyses informed by deep mutational scanning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/121830v1?rss=1</link>
<description><![CDATA[
BackgroundThe evolution of protein-coding genes can be quantitatively modeled using phylogenetic methods. Recently, it has been shown that high-throughput experimental measurements of mutational effects made via deep mutational scanning can inform site-specific phylogenetic substitution models of gene evolution. However, there is currently no software tailored for such analyses.nnResultsWe describe software that efficiently performs phylogenetic analyses with substitution models informed by deep mutational scanning. This software, phydms, is [~]100-fold faster than existing programs that accommodate such substitution models. It can be used to compare the results of deep mutational scanning experiments to the selection on genes in nature. For instance, phydms enables rigorous comparison of how well different experiments on the same gene describe natural selection. It also enables the re-scaling of deep mutational scanning data to account for differences in the stringency of selection in the lab and nature. Finally, phydms can identify sites that are evolving differently in nature than expected from experiments in the lab.nnConclusionsThe phydms software makes it easy to use phylogenetic substitution models informed by deep mutational scanning experiments. As data from such experiments becomes increasingly widespread, phydms will facilitate quantitative comparison of the experimental results to the actual selection pressures shaping evolution in nature.
]]></description>
<dc:creator>Hilton, S. K.</dc:creator>
<dc:creator>Doud, M. B.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2017-03-29</dc:date>
<dc:identifier>doi:10.1101/121830</dc:identifier>
<dc:title><![CDATA[phydms: Software for phylogenetic analyses informed by deep mutational scanning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/125609v1?rss=1">
<title>
<![CDATA[
Comprehensive characterization of pediatric acute myeloid leukemia reveals novel molecular features and age-specific interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/125609v1?rss=1</link>
<description><![CDATA[
We present the molecular landscape of pediatric acute myeloid leukemia (AML), characterizing nearly 1,000 participants in Childrens Oncology Group (COG) AML trials. The COG/NCI TARGET AML initiative assessed cases by whole-genome, targeted DNA, mRNA, miRNA sequencing and CpG methylation profiling. Validated DNA variants revealed diverse, infrequent mutations with fewer than 40 genes mutated in >2% of cases. In contrast, somatic structural variants, including novel gene fusions and focal MBNL1, ZEB2, and ELF1 deletions, were disproportionately prevalent in young as compared to adult patients. Conversely, DNMT3A and TP53 mutations, common in adults, are conspicuously absent from virtually all pediatric cases. Novel GATA2, FLT3, and CBL mutations, recurrent MYC-ITD, NRAS, KRAS, and WT1 mutations are frequent in pediatric AML. Deletions, mutations, and promoter DNA hypermethylation convergently impact Wnt signaling, Polycomb repression, innate immune cell interactions, and a cluster of zinc finger genes associated with KMT2A rearrangements. These results highlight the need for, and facilitate the development of age-tailored targeted therapies for the treatment of pediatric AML.
]]></description>
<dc:creator>Bolouri, H.</dc:creator>
<dc:creator>Farrar, J. E.</dc:creator>
<dc:creator>Triche, T. J.</dc:creator>
<dc:creator>Ries, R. E.</dc:creator>
<dc:creator>Lim, E. L.</dc:creator>
<dc:creator>Alonzo, T. A.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Moore, R.</dc:creator>
<dc:creator>Mungall, A.</dc:creator>
<dc:creator>Marra, M. A.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Ma, X.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Guidry Auvil, J. M.</dc:creator>
<dc:creator>Davidsen, T. M.</dc:creator>
<dc:creator>Gesuwan, P.</dc:creator>
<dc:creator>Hermida, L. C.</dc:creator>
<dc:creator>Salhia, B.</dc:creator>
<dc:creator>Capone, S.</dc:creator>
<dc:creator>Ramsingh, G.</dc:creator>
<dc:creator>Zwaan, C. M.</dc:creator>
<dc:creator>Noort, S.</dc:creator>
<dc:creator>Piccolo, S.</dc:creator>
<dc:creator>Kolb, E. A.</dc:creator>
<dc:creator>Gamis, A. S.</dc:creator>
<dc:creator>Smith, M. A.</dc:creator>
<dc:creator>Gerhard, D. S.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:date>2017-06-13</dc:date>
<dc:identifier>doi:10.1101/125609</dc:identifier>
<dc:title><![CDATA[Comprehensive characterization of pediatric acute myeloid leukemia reveals novel molecular features and age-specific interactions]]></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/10.1101/136614v1?rss=1">
<title>
<![CDATA[
Mapping Tumor-Specific Expression QTLs In Impure Tumor Samples 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/136614v1?rss=1</link>
<description><![CDATA[
The study of gene expression quantitative trait loci (eQTL) is an effective approach to illuminate the functional roles of genetic variants. Computational methods have been developed for eQTL mapping using gene expression data from microarray or RNA-seq technology. Application of these methods for eQTL mapping in tumor tissues is problematic because tumor tissues are composed of both tumor and infiltrating normal cells (e.g. immune cells) and eQTL effects may vary between tumor and infiltrating normal cells. To address this challenge, we have developed a new method for eQTL mapping using RNA-seq data from tumor samples. Our method separately estimates the eQTL effects in tumor and infiltrating normal cells using both total expression and allele-specific expression (ASE). We demonstrate that our method controls type I error rate and has higher power than some alternative approaches. We applied our method to study RNA-seq data from The Cancer Genome Atlas and illustrated the similarities and differences of eQTL effects in tumor and normal cells.
]]></description>
<dc:creator>Wilson, D. R.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:creator>Ibrahim, J. G.</dc:creator>
<dc:date>2017-05-11</dc:date>
<dc:identifier>doi:10.1101/136614</dc:identifier>
<dc:title><![CDATA[Mapping Tumor-Specific Expression QTLs In Impure Tumor Samples]]></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/10.1101/143347v1?rss=1">
<title>
<![CDATA[
Biochemical Adaptations Of The Retina And Retinal Pigment Epithelium Support A Metabolic Ecosystem In The Vertebrate Eye 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/143347v1?rss=1</link>
<description><![CDATA[
Here we report multiple lines of evidence for a comprehensive model for retinal energy metabolism. Metabolic flux, locations of key enzymes and our finding that glucose enters the neural retina almost entirely through photoreceptors support a conceptually new model for retinal metabolism. In this model, glucose from the choroidal blood supply passes through the retinal pigment epithelium to the retina where photoreceptors convert it to lactate. Photoreceptors then export the lactate as fuel for the retinal pigment epithelium and for neighboring Muller glial cells. A key feature of this model is that aerobic glycolysis in photoreceptors produces lactate to suppress glycolysis in the neighboring retinal pigment epithelium. That enhances the flow of glucose to the retina by minimizing consumption of glucose within the retinal pigment epithelium. This framework for metabolic relationships in retina provides new insights into the underlying causes of retinal disease, age-related vision loss and metabolism-based therapies.
]]></description>
<dc:creator>Kanow, M. A.</dc:creator>
<dc:creator>Giarmarco, M. M.</dc:creator>
<dc:creator>Jankowski, C.</dc:creator>
<dc:creator>Tsantilas, K.</dc:creator>
<dc:creator>Engel, A. L.</dc:creator>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Linton, J. D.</dc:creator>
<dc:creator>Farnsworth, C. C.</dc:creator>
<dc:creator>Sloat, S. R.</dc:creator>
<dc:creator>Lindsay, K. J.</dc:creator>
<dc:creator>Parker, E. D.</dc:creator>
<dc:creator>Brockerhoff, S. E.</dc:creator>
<dc:creator>Sadilek, M.</dc:creator>
<dc:creator>Chao, J. R.</dc:creator>
<dc:creator>Hurley, J. B.</dc:creator>
<dc:date>2017-05-29</dc:date>
<dc:identifier>doi:10.1101/143347</dc:identifier>
<dc:title><![CDATA[Biochemical Adaptations Of The Retina And Retinal Pigment Epithelium Support A Metabolic Ecosystem In The Vertebrate Eye]]></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/10.1101/146977v1?rss=1">
<title>
<![CDATA[
HIV persistence during antiviral therapy occurs due to cellular proliferation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/146977v1?rss=1</link>
<description><![CDATA[
Antiretroviral therapy (ART) suppresses viral replication in people living with HIV. Yet, infected cells persist for decades on ART and viremia returns if ART is stopped. Persistence has been attributed to viral replication in an ART sanctuary and long-lived and1or proliferating latently infected cells. Using ecological methods and existing data, we infer that >99% of infected cells are members of clonal populations after one year of ART. We reconcile our results with observations from the first months of ART, demonstrating mathematically how a "fossil record" of historic HIV replication permits observed viral evolution even while most new infected cells arise from proliferation. Together, our results imply cellular proliferation generates a majority of infected cells during ART. Therefore, reducing proliferation could decrease the size of the HIV reservoir and help achieve a functional cure.
]]></description>
<dc:creator>Reeves, D. B.</dc:creator>
<dc:creator>Duke, E. R.</dc:creator>
<dc:creator>Wagner, T. A.</dc:creator>
<dc:creator>Palmer, S. E.</dc:creator>
<dc:creator>Spivak, A. M.</dc:creator>
<dc:creator>Schiffer, J. T.</dc:creator>
<dc:date>2017-06-07</dc:date>
<dc:identifier>doi:10.1101/146977</dc:identifier>
<dc:title><![CDATA[HIV persistence during antiviral therapy occurs due to cellular proliferation]]></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/10.1101/165639v1?rss=1">
<title>
<![CDATA[
Transcription start site profiling uncovers divergent transcription and enhancer-associated RNAs in Drosophila melanogaster 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/165639v1?rss=1</link>
<description><![CDATA[
BackgroundHigh-resolution transcription start site (TSS) mapping in D. melanogaster embryos and cell lines has revealed a rich and detailed landscape of both cis- and trans-regulatory elements and factors. However, TSS profiling has not been investigated in an orthogonal in vivo setting. Here, we present a comprehensive dataset that links TSS dynamics with nucleosome occupancy and gene expression in the wandering third instar larva, a developmental stage characterized by large-scale shifts in transcriptional programs in preparation for metamorphosis.nnResultsThe data recapitulate major regulatory classes of TSSs, based on peak width, promoter-proximal polymerase pausing, and cis-regulatory element density. We confirm the paucity of divergent transcription units in D. melanogaster, but also identify notable exceptions. Furthermore, we identify thousands of novel initiation events occurring at unannotated TSSs that can be classified into functional categories by their local density of histone modifications. Interestingly, a sub-class of these unannotated TSSs overlaps with functionally validated enhancer elements, consistent with a regulatory role for "enhancer RNAs" (eRNAs) in defining transcriptional programs that are important for animal development.nnConclusionsHigh-depth TSS mapping is a powerful strategy for identifying and characterizing low-abundance and/or low-stability RNAs. Global analysis of transcription initiation patterns in a developing organism reveals a vast number of novel initiation events that identify potential eRNAs as well as other non-coding transcripts critical for animal development.
]]></description>
<dc:creator>Meers, M. P.</dc:creator>
<dc:creator>Adelman, K.</dc:creator>
<dc:creator>Duronio, R. J.</dc:creator>
<dc:creator>Strahl, B. D.</dc:creator>
<dc:creator>McKay, D. J.</dc:creator>
<dc:creator>Matera, A. G.</dc:creator>
<dc:date>2017-07-19</dc:date>
<dc:identifier>doi:10.1101/165639</dc:identifier>
<dc:title><![CDATA[Transcription start site profiling uncovers divergent transcription and enhancer-associated RNAs in Drosophila melanogaster]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/166595v1?rss=1">
<title>
<![CDATA[
Expanding the Atlas of Functional Missense Variation for Human Genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/166595v1?rss=1</link>
<description><![CDATA[
Although we now routinely sequence human genomes, we can confidently identify only a fraction of the sequence variants that have a functional impact. Here we developed a deep mutational scanning framework that produces exhaustive maps for human missense variants by combining random codon-mutagenesis and multiplexed functional variation assays with computational imputation and refinement. We applied this framework to four proteins corresponding to six human genes: UBE2I (encoding SUMO E2 conjugase), SUMO1 (small ubiquitin-like modifier), TPK1 (thiamin pyrophosphokinase), and CALM1/2/3 (three genes encoding the protein calmodulin). The resulting maps recapitulate known protein features, and confidently identify pathogenic variation. Assays potentially amenable to deep mutational scanning are already available for 57% of human disease genes, suggesting that DMS could ultimately map functional variation for all human disease genes.
]]></description>
<dc:creator>Weile, J.</dc:creator>
<dc:creator>Sun, S.</dc:creator>
<dc:creator>Cote, A. G.</dc:creator>
<dc:creator>Knapp, J.</dc:creator>
<dc:creator>Verby, M.</dc:creator>
<dc:creator>Mellor, J. C.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Pons, C.</dc:creator>
<dc:creator>Wong, C.</dc:creator>
<dc:creator>van Lieshout, N.</dc:creator>
<dc:creator>Yang, F.</dc:creator>
<dc:creator>Tasan, M.</dc:creator>
<dc:creator>Tan, G.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Fowler, D. M.</dc:creator>
<dc:creator>Nussbaum, R.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Vidal, M.</dc:creator>
<dc:creator>Hill, D. E.</dc:creator>
<dc:creator>Aloy, P.</dc:creator>
<dc:creator>Roth, F. P.</dc:creator>
<dc:date>2017-07-27</dc:date>
<dc:identifier>doi:10.1101/166595</dc:identifier>
<dc:title><![CDATA[Expanding the Atlas of Functional Missense Variation for Human Genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/167445v1?rss=1">
<title>
<![CDATA[
beachmat: a Bioconductor C++ API for accessing single-cell genomics data from a variety of R matrix types 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/167445v1?rss=1</link>
<description><![CDATA[
Recent advances in single-cell RNA sequencing have dramatically increased the number of cells that can be profiled in a single experiment. This provides unparalleled resolution to study cellular heterogeneity within biological processes such as differentiation. However, the explosion of data that are generated from such experiments poses a challenge to the existing computational infrastructure for statistical data analysis. In particular, large matrices holding expression values for each gene in each cell require sparse or file-backed representations for manipulation with the popular R programming language. Here, we describe a C++ interface named beachmat, which enables agnostic data access from various matrix representations. This allows package developers to write efficient C++ code that is interoperable with simple, sparse and HDF5-backed matrices, amongst others. We perform simulations to examine the performance of beachmat on each matrix representation, and we demonstrate how beachmat can be incorporated into the code of other packages to drive analyses of a very large single-cell data set.
]]></description>
<dc:creator>Lun, A. T. L.</dc:creator>
<dc:creator>Pages, H.</dc:creator>
<dc:creator>Smith, M. L.</dc:creator>
<dc:date>2017-07-24</dc:date>
<dc:identifier>doi:10.1101/167445</dc:identifier>
<dc:title><![CDATA[beachmat: a Bioconductor C++ API for accessing single-cell genomics data from a variety of R matrix types]]></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/10.1101/191676v1?rss=1">
<title>
<![CDATA[
Seasonal influenza circulation patterns and projections for Sep 2017 to Sep 2018 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/191676v1?rss=1</link>
<description><![CDATA[
This report details current seasonal influenza circulation patterns as of Sep 2017 and makes projections up to Sep 2018 to coincide with selection of the 2018 Southern Hemisphere vaccine strain. This is not meant as a comprehensive report, but is instead intended as particular observations that weve made that may be of relevance. Please also note that observed patterns reflect the GISAID database and may not be entirely representative of underlying dynamics. All analyses are based on the nextflu pipeline [1] with continual updates posted to nextflu.org.nnA/H3N2H3N2 continues to diversify with many coexisting clades, all of which carry several amino acid mutations at previously characterized epitope sites. The majority of viruses fall into the 3c2.a clade which has been dominating globally for >3 years, but 3c3.a viruses continue to persist. The common ancestor of circulating H3N2 viruses is now more than 5 years old, which is rare for H3N2. Despite extensive genetic diversity, serological assays suggest limited, but non-zero, antigenic evolution. We expect multiple competing clades within 3c2.a to persist into the future with no clear immediate winner.nnA/H1N1pdmA clade comprising mutations S74R and I295V has recently risen to >60% global frequency. Although it shows no antigenic distinction by ferret HI data, the rapidity of its rise suggests a selective origin.nnB/VicA clade with a two amino acid deletion 162-/163-has altered serological properties and is increasing in frequency, albeit slowly. Two other clades (carrying mutations K209N and V87A/I175V) have increased in frequency moderately.nnB/YamA clade comprising M251V within clade 3 viruses continues to dominate. The is little genetic differentiation within this clade and no evidence of antigenic evolution.
]]></description>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Neher, R.</dc:creator>
<dc:date>2017-09-26</dc:date>
<dc:identifier>doi:10.1101/191676</dc:identifier>
<dc:title><![CDATA[Seasonal influenza circulation patterns and projections for Sep 2017 to Sep 2018]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/193995v1?rss=1">
<title>
<![CDATA[
Extreme heterogeneity of influenza virus infection in single cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/193995v1?rss=1</link>
<description><![CDATA[
Viral infection can dramatically alter a cells transcriptome. However, these changes have mostly been studied by bulk measurements on many cells. Here we use single-cell mRNA sequencing to examine the transcriptional consequences of Influenza virus infection. We 1nd extremely wide cell-to-cell variation in production of viral gene transcripts - viral transcripts compose less than a percent of total mRNA in many infected cells, but a few cells derive over half their mRNA from virus. Some infected cells fail to express at least one viral gene, and this gene absence partially explains variation in viral transcriptional load. Despite variation in total viral load, the relative abundances of viral mRNAs are fairly consistent across infected cells. Activation of innate immune pathways is rare, but some cellular genes co-vary in abundance with the amount of viral mRNA. Overall, our results highlight the complexity of viral infection at the level of single cells.
]]></description>
<dc:creator>Russell, A. B.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2017-09-27</dc:date>
<dc:identifier>doi:10.1101/193995</dc:identifier>
<dc:title><![CDATA[Extreme heterogeneity of influenza virus infection in single cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.11.872986v1?rss=1">
<title>
<![CDATA[
A high-throughput screen for transcription activation domains reveals their sequence characteristics and permits reliable prediction by deep learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.11.872986v1?rss=1</link>
<description><![CDATA[
Transcription activation domains (ADs) are encoded by a wide range of seemingly unrelated amino acid sequences, making it difficult to recognize features that permit their dynamic behavior, fuzzy interactions and target specificity. We screened a large set of random 30-mer peptides for AD function and trained a deep neural network (ADpred) on the AD-positive and negative sequences. ADpred correctly identifies known ADs within protein sequences and accurately predicts the consequences of mutations. We show that functional ADs are (1) located within intrinsically disordered regions with biased amino acid composition, (2) contain clusters of hydrophobic residues near acidic side chains, (3) are enriched or depleted for particular dipeptide sequences, and (4) have higher helical propensity than surrounding regions. Taken together, our findings fit the model of "fuzzy" binding through hydrophobic protein-protein interfaces, where activator-coactivator binding takes place in a dynamic hydrophobic environment rather than through combinations of sequence-specific interactions.
]]></description>
<dc:creator>Erijman, A.</dc:creator>
<dc:creator>Kozlowski, L.</dc:creator>
<dc:creator>Sohrabi-Jahromi, S.</dc:creator>
<dc:creator>Fishburn, J.</dc:creator>
<dc:creator>Warfield, L.</dc:creator>
<dc:creator>Schreiber, J.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:creator>Soeding, J.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:date>2019-12-12</dc:date>
<dc:identifier>doi:10.1101/2019.12.11.872986</dc:identifier>
<dc:title><![CDATA[A high-throughput screen for transcription activation domains reveals their sequence characteristics and permits reliable prediction by deep learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.18.881763v1?rss=1">
<title>
<![CDATA[
Wash and the WASH Regulatory Complex function in Nuclear Envelope budding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.18.881763v1?rss=1</link>
<description><![CDATA[
Nuclear envelope budding is a recently described phenomenon wherein large macromolecular complexes can be packaged inside the nucleus and be extruded through the nuclear membranes, completely bypassing nuclear pores. While factors have been identified both as cargos or actively involved in this process, much remains unknown about the molecules that generate the forces and membrane deformations which appear inherent. Using fluorescence and electron microscopy, biochemical and cell biological assays, and genetic perturbations in the Drosophila model, we identify Wash, its regulatory complex, and Arp2/3 as novel players in NE-budding. Surprisingly, Washs role in this process is bipotent and, independent of SHRC/Arp2/3, its perturbation disrupts the normal homotypic Lamin A/B meshworks that are necessary for NE-budding to occur. In addition to NE-budding emerging as important in additional cellular processes and organisms, its incredible similarity to herpesvirus egress suggests new avenues for exploration in both normal and disease biology.
]]></description>
<dc:creator>Verboon, J. M.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Decker, J. R.</dc:creator>
<dc:creator>Davidson, K. A.</dc:creator>
<dc:creator>Nandakumar, V.</dc:creator>
<dc:creator>Parkhurst, S. M.</dc:creator>
<dc:date>2019-12-19</dc:date>
<dc:identifier>doi:10.1101/2019.12.18.881763</dc:identifier>
<dc:title><![CDATA[Wash and the WASH Regulatory Complex function in Nuclear Envelope budding]]></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/10.1101/2019.12.30.890517v1?rss=1">
<title>
<![CDATA[
Single-Cell Map of Diverse Immune Phenotypes in the Metastatic Brain Tumor Microenvironment of Non Small Cell Lung Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.30.890517v1?rss=1</link>
<description><![CDATA[
Cancer immunotherapies have shown sustained clinical success in treating primary non-small-cell lung cancer (NSCLC). However, patients with brain metastasis are excluded from the trials because the brain is viewed traditionally as an immune-privileged organ. The composition and properties of tumor-infiltrating myeloid cells in metastatic brain tumors are mostly unknown. To depict the baseline landscape of the composition, gene signature, and functional states of these immune cells, we performed - single-cell RNA sequencing (scRNAseq) for 12,196cellsafter data preprocessing, including 2,241 immunecells from three surgically removed brain lesions of treatment-naive NSCLC patients. We found a lack of T lymphocyte infiltration and activation, as well as the vast expansion of tumor-associated macrophage(TAM) in the brain lesions of NSCLC patients. By comparing our scRNAseq dataset with published data from early and late-stage primary NSCLC tumors, we showed that this compromised T cell response is unique to brain lesions. We identified a unique alternative activation (M2) gene expression pattern of the TAM in the brain metastasis and a lack of known T cell co-stimulator expression. Accumulation of M2 polarized TAM may, therefore, cause the comprised anti-tumor T cell response in metastatic brain lesions. These findings can contribute to the design of new immunotherapy strategies for NSCLC patients with brain metastasis.
]]></description>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Dai, J.</dc:creator>
<dc:creator>Han, R.-R.</dc:creator>
<dc:creator>Dong, L.</dc:creator>
<dc:creator>Feng, D.</dc:creator>
<dc:creator>Zhu, G.</dc:creator>
<dc:creator>Guo, W.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Chao, M.</dc:creator>
<dc:creator>Jin, M.-Z.</dc:creator>
<dc:creator>Jin, S.-J.</dc:creator>
<dc:creator>Wei, D.-P.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:creator>Jin, W.-L.</dc:creator>
<dc:date>2019-12-30</dc:date>
<dc:identifier>doi:10.1101/2019.12.30.890517</dc:identifier>
<dc:title><![CDATA[Single-Cell Map of Diverse Immune Phenotypes in the Metastatic Brain Tumor Microenvironment of Non Small Cell Lung Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.10.902296v1?rss=1">
<title>
<![CDATA[
BAF facilitates interphase nuclear envelope repair through recruitment of nuclear transmembrane proteins. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.10.902296v1?rss=1</link>
<description><![CDATA[
Nuclear membrane rupture during interphase occurs in a variety of cell contexts, both healthy and pathological. Membrane ruptures can be rapidly repaired, but these mechanisms are still unclear. Here we show BAF, a nuclear envelope protein that shapes chromatin and recruits membrane proteins in mitosis, also facilitates nuclear membrane repair in interphase, in part through recruitment of the nuclear membrane proteins emerin and LEMD2 to rupture sites. Characterization of GFP-BAF accumulation at nuclear membrane rupture sites confirmed BAF is a fast, accurate, and persistent mark of nucleus rupture whose kinetics are partially dictated by membrane resealing. BAF depletion significantly delayed nuclear membrane repair, with a larger effect on longer ruptures. This phenotype could be rescued by GFP-BAF, but not by a BAF mutant lacking the LEM-protein binding domain. Depletion of the BAF interactors LEMD2 or emerin, and to a lesser extent lamin A/C, increased the duration of nucleus ruptures, consistent with LEM-protein binding being a key function of BAF during membrane repair. Overall our results suggest a model where BAF is critical for timely repair of large ruptures in the nuclear membrane, potentially by facilitating membrane attachment to the rupture site.
]]></description>
<dc:creator>Young, A. M.</dc:creator>
<dc:creator>Gunn, A. L.</dc:creator>
<dc:creator>Hatch, E. M.</dc:creator>
<dc:date>2020-01-11</dc:date>
<dc:identifier>doi:10.1101/2020.01.10.902296</dc:identifier>
<dc:title><![CDATA[BAF facilitates interphase nuclear envelope repair through recruitment of nuclear transmembrane proteins.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.15.906867v1?rss=1">
<title>
<![CDATA[
Individual yeast cells signal at different levels but each with good precision 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.15.906867v1?rss=1</link>
<description><![CDATA[
Different isogenic cells have been shown to exhibit widely varying responses to the same extracellular signal. Based on the assumption that this variation arises from noise in the signaling pathways that cells use to transmit information from surface to nucleus, recent publications asserted that single cells cannot detect their surroundings accurately. Here, we analyze existing data on gene expression induced by the Saccharomyces cerevisiae pheromone response system, finding that individual cells signal consistently over time, implying that response variation arises primarily from stable cell-to-cell differences rather than signaling noise. Individual cells transmit at least 2.7 bits of information through the pheromone response system, enabling each cell to distinguish between at least 6 pheromone concentrations. In principle, cells can gain further precision by internally referencing these responses with measurements of constitutively expressed genes. Combination with prior results shows that only about 6% of total response variation arises from signaling pathway noise.

One-sentence summarySingle yeast cells signal consistently over time, indicating that their signaling pathways transmit information precisely.
]]></description>
<dc:creator>Andrews, S. S.</dc:creator>
<dc:creator>Brent, R.</dc:creator>
<dc:date>2020-01-15</dc:date>
<dc:identifier>doi:10.1101/2020.01.15.906867</dc:identifier>
<dc:title><![CDATA[Individual yeast cells signal at different levels but each with good precision]]></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/10.1101/2020.01.17.909937v1?rss=1">
<title>
<![CDATA[
Mechanical Instability of Adherens Junctions Overrides Intrinsic Quiescence of Hair Follicle Stem Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.17.909937v1?rss=1</link>
<description><![CDATA[
Vinculin, a mechanotransducer associated with both adherens junctions (AJ) and focal adhesions (FA) plays a central role in force transmission through these cell-cell and cell-substratum contacts. Here we describe the conditional knock out (KO) of vinculin in murine skin. Remarkably, we find that the loss of vinculin function results in the loss of bulge stem cell (BuSC) quiescence. We demonstrate that vinculin KO cells are impaired in force generation resulting in mechanically weak AJs. Mechanistically, vinculin functions by keeping -catenin in a stretched conformation, which in turn regulates the retention of YAP1, another potent mechanotransducer and regulator of cell proliferation, to the junctions. Conditional KO of -catenin specifically in the BuSCs further corroborates the importance of stable AJs in the maintenance of quiescence and stemness. Altogether, our data provides definitive mechanistic insights into the hitherto unexplored regulatory link between the mechanical stability of cell-junctions and the maintenance of BuSC quiescence.
]]></description>
<dc:creator>Biswas, R.</dc:creator>
<dc:creator>Banerjee, A.</dc:creator>
<dc:creator>Lembo, S.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Lakshmanan, V.</dc:creator>
<dc:creator>Nakasaki, M.</dc:creator>
<dc:creator>Kutyavin, V.</dc:creator>
<dc:creator>Wright, G.</dc:creator>
<dc:creator>Palakodeti, D.</dc:creator>
<dc:creator>Ross, R.</dc:creator>
<dc:creator>Jamora, C.</dc:creator>
<dc:creator>Vasioukhin, V.</dc:creator>
<dc:creator>Yan, J.</dc:creator>
<dc:creator>Raghavan, S.</dc:creator>
<dc:date>2020-01-17</dc:date>
<dc:identifier>doi:10.1101/2020.01.17.909937</dc:identifier>
<dc:title><![CDATA[Mechanical Instability of Adherens Junctions Overrides Intrinsic Quiescence of Hair Follicle Stem Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.23.917484v1?rss=1">
<title>
<![CDATA[
Estimation of Full-Length TprK Diversity in Treponema pallidum subspecies pallidum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.23.917484v1?rss=1</link>
<description><![CDATA[
Immune evasion and disease progression of Treponema pallidum subspecies pallidum are associated with sequence diversity in the hypervariable, putative outer membrane protein TprK. Previous attempts to study variation within TprK have sequenced at depths insufficient to fully appreciate the hypervariable nature of the protein, failed to establish linkage between the proteins 7 variable regions, or were conducted on strains passed through rabbits. As a consequence, a complete profiling of tprK during infection in the human host is still lacking. Furthermore, prior studies examining how T. pallidum uses its repertoire of genomic donor sites to generate diversity within the V regions of the tprK also yielded a partial understanding of this process, due to the limited number of tprK alleles examined. In this study, we used short- and long-read deep sequencing to directly characterize full-length tprK alleles from T. pallidum collected from early lesions of patients attending two STD clinics in Italy. Our data, combined with recent data available on Chinese T. pallidum strains, show the near complete absence of overlap in TprK sequences among the 41 strains profiled to date. Moreover, our data allowed us to redefine the boundaries of tprK V regions, identify 55 donor sites, and estimate the total number of TprK variants that T. pallidum can potentially generate. Altogether, our results support how T. pallidum TprK antigenic variation system is an unsurmountable obstacle for the human immune system to naturally achieve infection eradication, and reiterate the importance of this mechanism for pathogen persistence in the host.

ImportanceSyphilis continues to be a significant public health issue in both low- and high-income nations, including the United States, where the number of infectious syphilis cases has increased dramatically over the past five years. T. pallidum, the causative agent of syphilis, encodes an outer membrane protein TprK that undergoes segmental gene conversion to constantly create new sequences. We performed deep TprK profiling to understand full-length TprK diversity in T. pallidum-positive clinical specimens and compared these to all samples for which TprK deep sequencing is available. We found almost no overlap in TprK sequences between different patients. We further estimate that the total baseline junctional diversity of full-length TprK rivals that of current estimates of the human adaptive immune system. These data underscore the immunoevasive ability of TprK that allows T. pallidum to establish lifelong infection.
]]></description>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Lin, M. J.</dc:creator>
<dc:creator>Phung, Q.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Ciccarese, G.</dc:creator>
<dc:creator>Dal Conte, I.</dc:creator>
<dc:creator>Cusini, M.</dc:creator>
<dc:creator>Drago, F.</dc:creator>
<dc:creator>Giacani, L.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:date>2020-01-24</dc:date>
<dc:identifier>doi:10.1101/2020.01.23.917484</dc:identifier>
<dc:title><![CDATA[Estimation of Full-Length TprK Diversity in Treponema pallidum subspecies pallidum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.03.931972v1?rss=1">
<title>
<![CDATA[
Continual inactivation of stem cell functional identity genes stabilizes progenitor commitment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.03.931972v1?rss=1</link>
<description><![CDATA[
Expansion of the pool of stem cells that indirectly generate differentiated cells through intermediate progenitors drives vertebrate brain evolution. Due to a lack of lineage information, mechanistic investigation of the competency of stem cells to generate intermediate progenitors remains impossible. Fly larval brain neuroblasts provide excellent in vivo models for investigating the regulation of stem cell functionality during neurogenesis. Type II neuroblasts undergo indirect neurogenesis by dividing asymmetrically to generate a neuroblast and a progeny that commits to an intermediate progenitor (INP) identity. We identified Tailless (Tll) as the master regulator that maintains type II neuroblast functional identity, including the competency to generate INPs. Successive inactivation during INP commitment inhibits tll activation by Notch, preventing INPs from reacquiring neuroblast functionality. We propose that the continual inactivation of neural stem cell functional identity genes by histone deacetylation allows intermediate progenitors to stably commit to generating diverse differentiated cells during indirect neurogenesis.
]]></description>
<dc:creator>Quinto-Rives, N.</dc:creator>
<dc:creator>Komori, H.</dc:creator>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Kondo, S.</dc:creator>
<dc:creator>Dai, Q.</dc:creator>
<dc:creator>Moore, A. W.</dc:creator>
<dc:creator>Lee, C.-Y.</dc:creator>
<dc:date>2020-02-03</dc:date>
<dc:identifier>doi:10.1101/2020.02.03.931972</dc:identifier>
<dc:title><![CDATA[Continual inactivation of stem cell functional identity genes stabilizes progenitor commitment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.07.939439v1?rss=1">
<title>
<![CDATA[
Polymorphisms in human APOBEC3H differentially regulate ubiquitination and antiviral activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.07.939439v1?rss=1</link>
<description><![CDATA[
The APOBEC3 family of cytidine deaminases are an important part of the host innate immune defense against endogenous retroelements and retroviruses like human immunodeficiency virus (HIV). APOBEC3H (A3H) is the most polymorphic of the human APOBEC3 genes, with four major haplotypes circulating in the population. Haplotype II is the only antivirally-active variant of A3H, while the majority of the population possess independently destabilizing polymorphisms present in haplotype I (R105G) and haplotypes III and IV (N15del). Here, we show that instability introduced by either polymorphism is positively correlated with degradative ubiquitination, while haplotype II is protected from this modification. Inhibiting ubiquitination by mutating all of the A3H lysines increased expression of haplotypes III and IV, but these stabilized forms of haplotype III and IV had a strict nuclear localization, and did not incorporate into virions, nor exhibit antiviral activity, thus separating stabilization from function. On the other hand, the instability and functional deficiencies of haplotype III could be rescued by fusion to haplotype II, supporting a model by which antiviral A3H is actively stabilized through a cytoplasmic retention mechanism. Thus, the evolutionary loss of A3H activity in many humans involves functional deficiencies independent of protein stability.
]]></description>
<dc:creator>Chesarino, N. M.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2020-02-09</dc:date>
<dc:identifier>doi:10.1101/2020.02.07.939439</dc:identifier>
<dc:title><![CDATA[Polymorphisms in human APOBEC3H differentially regulate ubiquitination and antiviral activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.11.927475v1?rss=1">
<title>
<![CDATA[
Optimal timing for cancer screening and adaptive surveillance using mathematical modeling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.11.927475v1?rss=1</link>
<description><![CDATA[
Cancer screening and early detection efforts have been partially successful in reducing incidence and mortality but many improvements are needed. Although current medical practice is mostly informed by epidemiological studies, the decisions for guidelines are ultimately made ad hoc. We propose that quantitative optimization of protocols can potentially increase screening success and reduce overdiagnosis. Mathematical modeling of the stochastic process of cancer evolution can be used to derive and to optimize the timing of clinical screens so that the probability is maximal that a patient is screened within a certain "window of opportunity" for intervention when early cancer development may be observable. Alternative to a strictly empirical approach, or microsimulations of a multitude of possible scenarios, biologically-based mechanistic modeling can be used for predicting when best to screen and begin adaptive surveillance. We introduce a methodology for optimizing screening, assessing potential risks, and quantifying associated costs to healthcare using multiscale models. As a case study in Barretts esophagus (BE), we applied our methods for a model of esophageal adenocarcinoma (EAC) that was previously calibrated to US cancer registry data. We found optimal screening ages for patients with symptomatic gastroesophageal reflux disease to be older (58 for men, 64 for women) than what is currently recommended (age > 50 years). These ages are in a cost-effective range to start screening and were independently validated by data used in current guidelines. Our framework captures critical aspects of cancer evolution within BE patients for a more personalized screening design.

SignificanceOur study demonstrates how mathematical modeling of cancer evolution can be used to optimize screening regimes. Surveillance regimes could also be improved if they were based on these models.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=66 SRC="FIGDIR/small/927475v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Curtius, K.</dc:creator>
<dc:creator>Dewanji, A.</dc:creator>
<dc:creator>Hazelton, W. D.</dc:creator>
<dc:creator>Rubenstein, J. H.</dc:creator>
<dc:creator>Luebeck, E. G.</dc:creator>
<dc:date>2020-02-11</dc:date>
<dc:identifier>doi:10.1101/2020.02.11.927475</dc:identifier>
<dc:title><![CDATA[Optimal timing for cancer screening and adaptive surveillance using mathematical modeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.11.941351v1?rss=1">
<title>
<![CDATA[
A multilayered post-GWAS assessment on genetic susceptibility to pancreatic cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.11.941351v1?rss=1</link>
<description><![CDATA[
Pancreatic cancer (PC) is a complex disease in which both non-genetic and genetic factors interplay. To-date, 40 GWAS hits have been associated with PC risk in individuals of European descent, explaining 4.1% of the phenotypic variance. Here, we complemented a classical new PC GWAS (1D) with spatial autocorrelation analysis (2D) and Hi-C maps (3D) to gain additional insight into the inherited basis of PC. In-silico functional analysis of public genomic information allowed prioritization of potentially relevant candidate variants. We replicated 17/40 previous PC-GWAS hits and identified novel variants with potential biological functions. The spatial autocorrelation approach prioritized low MAF variants not detected by GWAS. These were further expanded via 3D interactions to 54 target regions with high functional relevance. This multi-step strategy, combined with an in-depth in-silico functional analysis, offers a comprehensive approach to advance the study of PC genetic susceptibility and could be applied to other diseases.
]]></description>
<dc:creator>Lopez de Maturana, E.</dc:creator>
<dc:creator>Rodriguez, J.</dc:creator>
<dc:creator>Alonso, L.</dc:creator>
<dc:creator>Lao, O.</dc:creator>
<dc:creator>Molina-Montes, E.</dc:creator>
<dc:creator>Martin-Antoniano, I.</dc:creator>
<dc:creator>Gomez-Rubio, P.</dc:creator>
<dc:creator>Lawlor, R.</dc:creator>
<dc:creator>Carrato, A.</dc:creator>
<dc:creator>Hidalgo, M.</dc:creator>
<dc:creator>Iglesias, M.</dc:creator>
<dc:creator>Molero, X.</dc:creator>
<dc:creator>Lohr, M.</dc:creator>
<dc:creator>Michalski, C.</dc:creator>
<dc:creator>Perea, J.</dc:creator>
<dc:creator>O'Rorke, M.</dc:creator>
<dc:creator>Barbera, V.</dc:creator>
<dc:creator>Tardon, A.</dc:creator>
<dc:creator>Farre, A.</dc:creator>
<dc:creator>Munoz-Bellvis, L.</dc:creator>
<dc:creator>Crnogorac-Jurcevic, T.</dc:creator>
<dc:creator>Dominguez-Munoz, E.</dc:creator>
<dc:creator>Gress, T.</dc:creator>
<dc:creator>Greenhaf, W.</dc:creator>
<dc:creator>Sharp, L.</dc:creator>
<dc:creator>Arnes, L.</dc:creator>
<dc:creator>Cecchini, L.</dc:creator>
<dc:creator>Balsells, J.</dc:creator>
<dc:creator>Costello, E.</dc:creator>
<dc:creator>Ilzarbe, L.</dc:creator>
<dc:creator>Kleeff, J.</dc:creator>
<dc:creator>Kong, B.</dc:creator>
<dc:creator>Marquez, M.</dc:creator>
<dc:creator>Mora, J.</dc:creator>
<dc:creator>O'Driscoll, D.</dc:creator>
<dc:creator>Scarpa, A.</dc:creator>
<dc:creator>Ye, W.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Investigators, P.</dc:creator>
<dc:creator>Garcia-Closas, M.</dc:creator>
<dc:creator>Kogevinas, M.</dc:creator>
<dc:creator>Rothman, N.</dc:creator>
<dc:creator>Silverman, D.</dc:creator>
<dc:date>2020-02-12</dc:date>
<dc:identifier>doi:10.1101/2020.02.11.941351</dc:identifier>
<dc:title><![CDATA[A multilayered post-GWAS assessment on genetic susceptibility to pancreatic cancer]]></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/10.1101/2020.02.12.944629v1?rss=1">
<title>
<![CDATA[
Retrocopying expands the functional repertoire of APOBEC3 antiviral proteins in primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.12.944629v1?rss=1</link>
<description><![CDATA[
Host-virus arms races are inherently asymmetric; viruses evolve much more rapidly than host genomes. Thus, there is high interest in discovering mechanisms by which host genomes keep pace with rapidly evolving viruses. One family of restriction factors, the APOBEC3 (A3) cytidine deaminases, has undergone positive selection and expansion via segmental gene duplication and recombination. Here, we show that new copies of A3 genes have also been created in primates by reverse transcriptase-encoding elements like LINE-1 or endogenous retroviruses via a process termed retrocopying. First, we discovered that all simian primate genomes retain the remnants of an ancient A3 retrocopy: A3I. Furthermore, we found that some New World monkeys encode up to ten additional APOBEC3G (A3G) retrocopies. Some of these A3G retrocopies are transcribed in a variety of tissues and able to restrict retroviruses. Our findings suggest that host genomes co-opt retroelement activity in the germline to create new host restriction factors as another means to keep pace with the rapid evolution of viruses. (163)
]]></description>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:creator>McLaughlin, R. N.</dc:creator>
<dc:date>2020-02-13</dc:date>
<dc:identifier>doi:10.1101/2020.02.12.944629</dc:identifier>
<dc:title><![CDATA[Retrocopying expands the functional repertoire of APOBEC3 antiviral proteins in primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.16.950378v1?rss=1">
<title>
<![CDATA[
Proteogenomic heterogeneity of localized human prostate cancer progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.16.950378v1?rss=1</link>
<description><![CDATA[
BackgroundTumor-specific genomic aberrations are routinely determined by high throughput genomic measurements. It remains unclear though, how complex genome alterations affect molecular networks through changing protein levels, and consequently biochemical states of tumor tissues.

ResultsHere, we investigated the propagation of genomic effects along the axis of gene expression during prostate cancer progression. For that, we quantified genomic, transcriptomic and proteomic alterations based on 105 prostate samples, consisting of benign prostatic hyperplasia regions and malignant tumors, from 39 prostate cancer patients. Our analysis revealed convergent effects of distinct copy number alterations impacting on common downstream proteins, which are important for establishing the tumor phenotype. We devised a network-based approach that integrates perturbations across different molecular layers, which identified a sub-network consisting of nine genes whose joint activity positively correlated with increasingly aggressive tumor phenotypes and was predictive of recurrence-free survival. Further, our data revealed a wide spectrum of intra-patient network effects, ranging from similar to very distinct alterations on different molecular layers.

ConclusionsThis study uncovered molecular networks with remarkably convergent alterations across tumor sites and patients, but it also exposed a diversity of network effects: we could not identify a single sub-network that was perturbed in all high-grade tumor regions.
]]></description>
<dc:creator>Charmpi, K.</dc:creator>
<dc:creator>Guo, T.</dc:creator>
<dc:creator>Zhong, Q.</dc:creator>
<dc:creator>Wagner, U.</dc:creator>
<dc:creator>Sun, R.</dc:creator>
<dc:creator>Toussaint, N. C.</dc:creator>
<dc:creator>Fritz, C.</dc:creator>
<dc:creator>Yuan, C.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Rupp, N. J.</dc:creator>
<dc:creator>Christiansen, A.</dc:creator>
<dc:creator>Rutishauser, D.</dc:creator>
<dc:creator>Ruschoff, J. H.</dc:creator>
<dc:creator>Fankhauser, C.</dc:creator>
<dc:creator>Saba, K.</dc:creator>
<dc:creator>Poyet, C.</dc:creator>
<dc:creator>Hermanns, T.</dc:creator>
<dc:creator>Oehl, K.</dc:creator>
<dc:creator>Moore, A. L.</dc:creator>
<dc:creator>Beisel, C.</dc:creator>
<dc:creator>Calzone, L.</dc:creator>
<dc:creator>MARTIGNETTI, L.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Zhu, Y. J.</dc:creator>
<dc:creator>Martinez, M. R.</dc:creator>
<dc:creator>Manica, M.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Aebersold, R.</dc:creator>
<dc:creator>Wild, P. J.</dc:creator>
<dc:creator>Beyer, A.</dc:creator>
<dc:date>2020-02-16</dc:date>
<dc:identifier>doi:10.1101/2020.02.16.950378</dc:identifier>
<dc:title><![CDATA[Proteogenomic heterogeneity of localized human prostate cancer progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.19.956581v1?rss=1">
<title>
<![CDATA[
Structure, function and antigenicity of the SARS-CoV-2 spike glycoprotein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.19.956581v1?rss=1</link>
<description><![CDATA[
The recent emergence of a novel coronavirus associated with an ongoing outbreak of pneumonia (Covid-2019) resulted in infections of more than 72,000 people and claimed over 1,800 lives. Coronavirus spike (S) glycoprotein trimers promote entry into cells and are the main target of the humoral immune response. We show here that SARS-CoV-2 S mediates entry in VeroE6 cells and in BHK cells transiently transfected with human ACE2, establishing ACE2 as a functional receptor for this novel coronavirus. We further demonstrate that the receptor-binding domains of SARS-CoV-2 S and SARS-CoV S bind with similar affinities to human ACE2, which correlates with the efficient spread of SARS-CoV-2 among humans. We found that the SARS-CoV-2 S glycoprotein harbors a furin cleavage site at the boundary between the S1/S2 subunits, which is processed during biogenesis and sets this virus apart from SARS-CoV and other SARS-related CoVs. We determined a cryo-electron microscopy structure of the SARS-CoV-2 S ectodomain trimer, demonstrating spontaneous opening of the receptor-binding domain, and providing a blueprint for the design of vaccines and inhibitors of viral entry. Finally, we demonstrate that SARS-CoV S murine polyclonal sera potently inhibited SARS-CoV-2 S-mediated entry into target cells, thereby indicating that cross-neutralizing antibodies targeting conserved S epitopes can be elicited upon vaccination.
]]></description>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>Park, Y.-J.</dc:creator>
<dc:creator>Tortorici, M. A.</dc:creator>
<dc:creator>Wall, A.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:date>2020-02-20</dc:date>
<dc:identifier>doi:10.1101/2020.02.19.956581</dc:identifier>
<dc:title><![CDATA[Structure, function and antigenicity of the SARS-CoV-2 spike glycoprotein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.21.960484v1?rss=1">
<title>
<![CDATA[
Evolutionarily conserved regulation of embryonic fast-twitch skeletal muscle differentiation by Pbx factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.21.960484v1?rss=1</link>
<description><![CDATA[
Vertebrate skeletal muscles are composed of both slow-twitch and fast-twitch fiber types. How the differentiation of distinct fiber types is activated during embryogenesis is not well characterized. Skeletal muscle differentiation is initiated by the activity of the myogenic basic helix-loop-helix (bHLH) transcription factors Myf5, Myod1, Myf6, and Myog. Myod1 functions as a muscle master regulatory factor and directly activates muscle differentiation genes, including those specific to both slow and fast muscle fibers. Our previous studies showed that Pbx TALE-class homeodomain proteins bind with Myod1 on the promoter of the zebrafish fast muscle gene mylpfa and are required for proper activation of mylpfa expression and the fast-twitch muscle-specific differentiation program in zebrafish embryos. Pbx proteins have also been shown to bind regulatory regions of muscle differentiation genes in mammalian muscle cells in culture. Here, we use new zebrafish mutant strains to confirm the essential roles of zebrafish Pbx factors in embryonic fast muscle differentiation. Furthermore, we examine the requirements for Pbx genes in mouse embryonic skeletal muscle differentiation, an area that has not been investigated in the mammalian embryo. Removing Pbx1 function from skeletal muscle in Myf5Cre/+;Pbx1fl/fl mouse embryos has minor effects on embryonic muscle development. However, concomitantly deleting Pbx2 function in Myf5Cre/+;Pbx1fl/fl;Pbx2-/- mouse embryos causes delayed activation and reduced expression of fast muscle differentiation genes. In the mouse, Pbx1/Pbx2-dependent fast muscle genes closely match those that have been previously shown to be dependent on murine Six1 and Six4. This work establishes evolutionarily conserved requirements for Pbx factors in embryonic fast muscle differentiation. Our studies are revealing how Pbx homeodomain proteins help direct specific cellular differentiation pathways.
]]></description>
<dc:creator>Farr, G. H.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Risolino, M.</dc:creator>
<dc:creator>Johnson, N. M.</dc:creator>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Kao, R. M.</dc:creator>
<dc:creator>Majesky, M. W.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:creator>Selleri, L.</dc:creator>
<dc:creator>Maves, L.</dc:creator>
<dc:date>2020-02-22</dc:date>
<dc:identifier>doi:10.1101/2020.02.21.960484</dc:identifier>
<dc:title><![CDATA[Evolutionarily conserved regulation of embryonic fast-twitch skeletal muscle differentiation by Pbx factors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.08.978239v1?rss=1">
<title>
<![CDATA[
Single-cell transcriptomics of allo-reactive CD4+ T cells over time reveals divergent fates during gut GVHD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.08.978239v1?rss=1</link>
<description><![CDATA[
Acute gastrointestinal Graft-versus-Host-Disease (GVHD) is a primary determinant of mortality after allogeneic hematopoietic stem-cell transplantation (alloSCT). It is mediated by alloreactive donor CD4+ T cells that differentiate into pathogenic subsets expressing IFN{gamma}, IL-17A or GM-CSF, and is regulated by subsets expressing IL-10 and/or Foxp3. Developmental relationships between T-helper states during priming in mesenteric lymph nodes (mLN) and effector function in the GI tract remain undefined at genome-scale. We used scRNA-seq and computational modelling to create an atlas of putative differentiation pathways during GVHD. Computational trajectory inference suggested emergence of pathogenic and regulatory states along a single developmental trajectory in mLN. Importantly, we identified an unexpected second trajectory, categorised by little proliferation or cytokine expression, reduced glycolysis, and high TCF1 expression. TCF1hi cells upregulated 4{beta}7 prior to gut migration and failed to express cytokines therein. Nevertheless, they demonstrated recall potential and plasticity following secondary transplantation, including cytokine or Foxp3 expression, but reduced TCF1. Thus, scRNA-seq revealed divergence of allo-reactive CD4+ T cells into quiescent and effector states during gut GVHD, reflecting putative heterogenous priming in vivo. These findings, the first at a single-cell level during GVHD over time, can now be used to interrogate T cell differentiation in patients undergoing alloSCT.
]]></description>
<dc:creator>Engel, J. A.</dc:creator>
<dc:creator>Lee, H. J.</dc:creator>
<dc:creator>Williams, C. G.</dc:creator>
<dc:creator>Kuns, R.</dc:creator>
<dc:creator>Olver, S.</dc:creator>
<dc:creator>Lansink, L. I. M.</dc:creator>
<dc:creator>Soon, M. S. F.</dc:creator>
<dc:creator>Andersen, S. B.</dc:creator>
<dc:creator>Powell, J. E.</dc:creator>
<dc:creator>Svensson, V.</dc:creator>
<dc:creator>Teichmann, S. A.</dc:creator>
<dc:creator>Hill, G. R.</dc:creator>
<dc:creator>Varelias, A.</dc:creator>
<dc:creator>Koyama, M.</dc:creator>
<dc:creator>Haque, A.</dc:creator>
<dc:date>2020-03-09</dc:date>
<dc:identifier>doi:10.1101/2020.03.08.978239</dc:identifier>
<dc:title><![CDATA[Single-cell transcriptomics of allo-reactive CD4+ T cells over time reveals divergent fates during gut GVHD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.11.978387v1?rss=1">
<title>
<![CDATA[
A single cell atlas reveals distinct immune landscapes in transplant and primary tumors that determine response or resistance to immunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.11.978387v1?rss=1</link>
<description><![CDATA[
Despite impressive responses in some patients, immunotherapy fails to cure most cancer patients. Preclinical studies indicate that radiotherapy synergizes with immunotherapy, promoting radiation-induced antitumor immunity. Nearly all preclinical immunotherapy studies utilize transplant tumor models, but cure rates of transplant tumor models treated with immunotherapy often overestimate patient responses. Here, we show that transplant tumors are cured by PD-1 blockade and radiotherapy, but identical treatment fails in autochthonous tumors. We generated a single-cell atlas of tumor-infiltrating immune cells from transplant and primary tumors treated with radiation and immunotherapy, which reveals striking differences in their immune landscapes. Although radiotherapy remodels myeloid cell phenotypes in primary and transplant tumors, only transplant tumors are enriched for CD8+ T cells that mediate tumor clearance while mice with primary sarcomas demonstrate tumor-specific tolerance. These results identify distinct microenvironments in tumors that coevolve with the immune system, which promote tolerance that must be overcome for immune-mediated cancer cures.
]]></description>
<dc:creator>Wisdom, A. J.</dc:creator>
<dc:creator>Mowery, Y. M.</dc:creator>
<dc:creator>Hong, C. S.</dc:creator>
<dc:creator>Qin, X.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Himes, J. E.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Fradin, H.</dc:creator>
<dc:creator>Muise, E. S.</dc:creator>
<dc:creator>Xu, E. S.</dc:creator>
<dc:creator>Carpenter, D. J.</dc:creator>
<dc:creator>Kent, C. L.</dc:creator>
<dc:creator>Smythe, K. S.</dc:creator>
<dc:creator>Williams, N.</dc:creator>
<dc:creator>Luo, L.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Owzar, K.</dc:creator>
<dc:creator>Bradley, T.</dc:creator>
<dc:creator>Kirsch, D. G.</dc:creator>
<dc:date>2020-03-12</dc:date>
<dc:identifier>doi:10.1101/2020.03.11.978387</dc:identifier>
<dc:title><![CDATA[A single cell atlas reveals distinct immune landscapes in transplant and primary tumors that determine response or resistance to immunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.20.001008v1?rss=1">
<title>
<![CDATA[
RT-qPCR DETECTION OF SARS-CoV-2 RNA FROM PATIENT NASOPHARYNGEAL SWAB USING QIAGEN RNEASY KITS OR DIRECTLY VIA OMISSION OF AN RNA EXTRACTION STEP 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.20.001008v1?rss=1</link>
<description><![CDATA[
The ongoing COVID-19 pandemic has caused an unprecedented need for rapid diagnostic testing. The Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO) recommend a standard assay that includes an RNA extraction step from a nasopharyngeal (NP) swab followed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to detect the purified SARS-CoV-2 RNA. The current global shortage of RNA extraction kits has caused a severe bottleneck to COVID-19 testing. We hypothesized that SARS-CoV-2 RNA could be detected from NP samples via a direct RT-qPCR assay that omits the RNA extraction step altogether, and tested this hypothesis on a series of blinded clinical samples. The direct RT-qPCR approach correctly identified 92% of NP samples (n = 155) demonstrated to be positive for SARS-CoV-2 RNA by traditional clinical diagnostic RT-qPCR that included an RNA extraction. Thus, direct RT-qPCR could be a front-line approach to identify the substantial majority of COVID-19 patients, reserving a repeat test with RNA extraction for those individuals with high suspicion of infection but an initial negative result. This strategy would drastically ease supply chokepoints of COVID-19 testing and should be applicable throughout the world.
]]></description>
<dc:creator>Bruce, E. A.</dc:creator>
<dc:creator>Tighe, S.</dc:creator>
<dc:creator>Hoffman, J. J.</dc:creator>
<dc:creator>Laaguiby, P.</dc:creator>
<dc:creator>Gerrard, D. L.</dc:creator>
<dc:creator>Diehl, S. A.</dc:creator>
<dc:creator>Leonard, D. G. B.</dc:creator>
<dc:creator>Huston, C. D.</dc:creator>
<dc:creator>Kirkpatrick, B. D.</dc:creator>
<dc:creator>Crothers, J. W.</dc:creator>
<dc:creator>Dragon, J.</dc:creator>
<dc:creator>Botten, J. W.</dc:creator>
<dc:date>2020-03-21</dc:date>
<dc:identifier>doi:10.1101/2020.03.20.001008</dc:identifier>
<dc:title><![CDATA[RT-qPCR DETECTION OF SARS-CoV-2 RNA FROM PATIENT NASOPHARYNGEAL SWAB USING QIAGEN RNEASY KITS OR DIRECTLY VIA OMISSION OF AN RNA EXTRACTION STEP]]></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/10.1101/2020.03.27.012963v1?rss=1">
<title>
<![CDATA[
APOBEC3C tandem domain proteins create super restriction factors against HIV-1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.27.012963v1?rss=1</link>
<description><![CDATA[
Humans encode proteins, called restriction factors, that inhibit replication of viruses like HIV-1. One family of antiviral proteins, apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (APOBEC3, shortened to A3) acts by deaminating cytidines to uridines during the reverse transcription reaction of HIV-1. The A3 locus encodes seven genes, named A3A-A3H. These genes either have one or two cytidine deaminase domains and several of these A3s potently restrict HIV-1. A3C, which has only a single cytidine deaminase domain, however, inhibits HIV-1 only very weakly. We tested novel double domain protein combinations by genetically linking two A3C genes to make a synthetic tandem domain protein. This protein created a "super restriction factor" that had more potent antiviral activity than the native A3C protein, which correlated with increased packaging into virions. Furthermore, disabling one of the active sites of the synthetic tandem domain protein results in an even greater increase in the antiviral activity--recapitulating a similar evolution seen in A3F and A3G (double domain A3s that only use a single catalytically active deaminase domain). These A3C tandem domain proteins do not have an increase in mutational activity, but instead inhibit formation of reverse transcription products which correlates with their ability to form large higher order complexes in cells. Finally, the A3C-A3C super restriction factor largely escaped antagonism by the HIV-1 viral protein, Vif.

ImportanceAs a part of the innate immune system, humans encode proteins that inhibit viruses like HIV-1. These broadly acting antiviral proteins do not protect humans from viral infections because viruses encode proteins that antagonize the host antiviral proteins to evade the innate immune system. One such example of a host antiviral protein is APOBEC3C (A3C), which weakly inhibits HIV-1. Here, we show that we can improve the antiviral activity of A3C by duplicating the DNA sequence to create a synthetic tandem domain, and furthermore, are relatively resistant to the viral antagonist, Vif. Together, these data give insights about how nature has evolved a defense against viral pathogens like HIV.
]]></description>
<dc:creator>McDonnell, M. M.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2020-03-29</dc:date>
<dc:identifier>doi:10.1101/2020.03.27.012963</dc:identifier>
<dc:title><![CDATA[APOBEC3C tandem domain proteins create super restriction factors against HIV-1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.03.023135v1?rss=1">
<title>
<![CDATA[
High-resolution influenza mapping of a city reveals socioeconomic determinants of transmission within and between urban quarters 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.03.023135v1?rss=1</link>
<description><![CDATA[
With two-thirds of the global population projected to be living in urban areas by 2050, understanding the transmission patterns of viral pathogens within cities is crucial for effective prevention strategies. Here, in unprecedented spatial resolution, we analysed the socioeconomic determinants of influenza transmission in a European city. We combined geographical and epidemiological data with whole genome sequencing of influenza viruses at the scale of urban quarters and statistical blocks, the smallest geographic subdivisions within a city. We observed annually re-occurring geographic clusters of influenza incidences, mainly associated with net income, and independent of population density and living space. Vaccination against influenza was also mainly associated with household income and was linked to the likelihood of influenza-like illness within an urban quarter. Transmissions patterns within and between quarters were complex. High-resolution city-level epidemiological studies combined with social science surveys such as this will be essential for understanding seasonal and pandemic transmission chains and delivering tailored public health information and vaccination programs at the municipal level.
]]></description>
<dc:creator>Egli, A.</dc:creator>
<dc:creator>Goldman, N.</dc:creator>
<dc:creator>Müller, N. F.</dc:creator>
<dc:creator>Brunner, M.</dc:creator>
<dc:creator>Wuethrich, D.</dc:creator>
<dc:creator>Tschudin-Sutter, S.</dc:creator>
<dc:creator>Hodcroft, E.</dc:creator>
<dc:creator>Neher, R.</dc:creator>
<dc:creator>Saalfrank, C.</dc:creator>
<dc:creator>Hadfield, J.</dc:creator>
<dc:creator>Bedfors, T.</dc:creator>
<dc:creator>Syedbasha, M.</dc:creator>
<dc:creator>Vogel, T.</dc:creator>
<dc:creator>Augustin, N.</dc:creator>
<dc:creator>Bauer, J.</dc:creator>
<dc:creator>Sailer, N.</dc:creator>
<dc:creator>Amar-Sliwad, N.</dc:creator>
<dc:creator>Lang, D.</dc:creator>
<dc:creator>Seth-Smith, H. M.</dc:creator>
<dc:creator>Blaich, A.</dc:creator>
<dc:creator>Hollenstein, Y.</dc:creator>
<dc:creator>Dubuis, O.</dc:creator>
<dc:creator>Naegele, M.</dc:creator>
<dc:creator>Buser, A.</dc:creator>
<dc:creator>Nickel, C.</dc:creator>
<dc:creator>Ritz, N.</dc:creator>
<dc:creator>Zeller, A.</dc:creator>
<dc:creator>Stadler, T.</dc:creator>
<dc:creator>Battegay, M.</dc:creator>
<dc:creator>Schneider-Sliwa, R.</dc:creator>
<dc:date>2020-04-04</dc:date>
<dc:identifier>doi:10.1101/2020.04.03.023135</dc:identifier>
<dc:title><![CDATA[High-resolution influenza mapping of a city reveals socioeconomic determinants of transmission within and between urban quarters]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.06.028662v1?rss=1">
<title>
<![CDATA[
Autocrine insulin pathway signaling regulates actin dynamics in cell wound repair 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.06.028662v1?rss=1</link>
<description><![CDATA[
Cells are exposed to frequent mechanical and/or chemical stressors that can compromise the integrity of the plasma membrane and underlying cortical cytoskeleton. The molecular mechanisms driving the immediate repair response launched to restore the cell cortex and circumvent cell death are largely unknown. Using microarrays and drug-inhibition studies to assess gene expression, we find that initiation of cell wound repair in the Drosophila model is dependent on translation, whereas transcription is required for subsequent steps. We identified 253 genes whose expression is up-regulated (80) or down-regulated (173) in response to laser wounding. A subset of these genes were validated using RNAi knockdowns and exhibit aberrant actomyosin ring assembly and/or actin remodeling defects. Strikingly, we find that the canonical insulin signaling pathway controls actin dynamics through the actin regulators Girdin and Chickadee (profilin), and its disruption leads to abnormal wound repair. Our results provide new insight for understanding how cell wound repair proceeds in healthy individuals and those with diseases involving wound healing deficiencies.
]]></description>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Dominguez, A. N. M.</dc:creator>
<dc:creator>Verboon, J. M.</dc:creator>
<dc:creator>Abreu-Blanco, M. T.</dc:creator>
<dc:creator>Liu, R.</dc:creator>
<dc:creator>Delrow, J. J.</dc:creator>
<dc:creator>Parkhurst, S. M.</dc:creator>
<dc:date>2020-04-07</dc:date>
<dc:identifier>doi:10.1101/2020.04.06.028662</dc:identifier>
<dc:title><![CDATA[Autocrine insulin pathway signaling regulates actin dynamics in cell wound repair]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.10.030460v1?rss=1">
<title>
<![CDATA[
Mapping the unique and shared functions of oncogenic KRAS and RIT1 with proteome and transcriptome profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.10.030460v1?rss=1</link>
<description><![CDATA[
Aberrant activation of RAS oncogenes is prevalent in lung adenocarcinoma, with somatic mutation of KRAS occurring in [~]30% of tumors. Recently, we identified somatic mutation of the RAS-family GTPase RIT1 in lung adenocarcinoma, but relatively little is known about the biological pathways regulated by RIT1 and how these relate to the oncogenic KRAS network. Here we present quantitative proteomic and transcriptomic profiles from KRAS-mutant and RIT1-mutant isogenic lung epithelial cells and globally characterize the signaling networks regulated by each oncogene. We find that both mutant KRAS and mutant RIT1 promote S6 kinase, AKT, and RAF/MEK signaling, and promote epithelial-to-mesenchymal transition and immune evasion via HLA protein loss. However, KRAS and RIT1 diverge in regulation of phosphorylation sites on EGFR, USO1, and AHNAK proteins. The majority of the proteome changes are related to altered transcriptional regulation, but a small subset of proteins are differentially regulated by both oncoproteins at the post-transcriptional level, including intermediate filament proteins, metallothioneins, and MHC Class I proteins. These data provide the first global, unbiased characterization of oncogenic RIT1 network and identify the shared and divergent functions of oncogenic RIT1 and KRAS GTPases in lung cancer.
]]></description>
<dc:creator>Lo, A.</dc:creator>
<dc:creator>Holmes, K.</dc:creator>
<dc:creator>Mundt, F.</dc:creator>
<dc:creator>Moorthi, S.</dc:creator>
<dc:creator>Fung, I.</dc:creator>
<dc:creator>Fereshetian, S.</dc:creator>
<dc:creator>Watson, J.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Mertins, P.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:date>2020-04-11</dc:date>
<dc:identifier>doi:10.1101/2020.04.10.030460</dc:identifier>
<dc:title><![CDATA[Mapping the unique and shared functions of oncogenic KRAS and RIT1 with proteome and transcriptome profiling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.14.041731v1?rss=1">
<title>
<![CDATA[
Chloroquine Kills Hair Cells in Zebrafish Lateral Line and Murine Cochlear Cultures: Implications for Ototoxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.14.041731v1?rss=1</link>
<description><![CDATA[
Hearing and balance deficits have been reported during and following treatment with the antimalarial drug chloroquine. However, experimental work examining the direct actions of chloroquine on mechanoreceptive hair cells in common experimental models is lacking. This study examines the effects of chloroquine on hair cells using two common experimental models: the zebrafish lateral line and neonatal mouse cochlear cultures. Zebrafish larvae were exposed to varying concentrations of chloroquine phosphate or hydroxychloroquine for 1 hr or 24 hr, and hair cells assessed by antibody staining. A significant, dose-dependent reduction in the number of surviving hair cells was seen across conditions for both exposure periods. Hydroxycholroquine showed similar toxicity. In mouse cochlear cultures, chloroquine damage was specific to outer hair cells in tissue from the cochlear basal turn, consistent with susceptibility to other ototoxic agents. These findings suggest a need for future studies employing hearing and balance monitoring during exposure to chloroquine and related compounds, particularly with interest in these compounds as therapeutics against viral infections including coronavirus.
]]></description>
<dc:creator>Davis, S. N.</dc:creator>
<dc:creator>Wu, P.</dc:creator>
<dc:creator>Camci, E. D.</dc:creator>
<dc:creator>Simon, J. A.</dc:creator>
<dc:creator>Rubel, E. W.</dc:creator>
<dc:creator>Raible, D. W.</dc:creator>
<dc:date>2020-04-16</dc:date>
<dc:identifier>doi:10.1101/2020.04.14.041731</dc:identifier>
<dc:title><![CDATA[Chloroquine Kills Hair Cells in Zebrafish Lateral Line and Murine Cochlear Cultures: Implications for Ototoxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.15.043083v1?rss=1">
<title>
<![CDATA[
Profiling the epigenome at home 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.15.043083v1?rss=1</link>
<description><![CDATA[
Chromatin accessibility mapping is a powerful approach to identify potential regulatory elements. A popular example is ATAC-seq, whereby Tn5 transposase inserts sequencing adapters into accessible DNA ( tagmentation). CUT&Tag is a tagmentation-based epigenomic profiling method in which antibody tethering of Tn5 to a chromatin epitope of interest profiles specific chromatin features in small samples and single cells. Here we show that by simply modifying the tagmentation conditions for histone H3K4me2 or H3K4me3 CUT&Tag, antibody-tethered tagmentation of accessible DNA sites is redirected to produce chromatin accessibility maps that are indistinguishable from the best ATAC-seq maps. Thus, chromatin accessibility maps can be produced in parallel with CUT&Tag maps of other epitopes with all steps from nuclei to amplified sequencing-ready libraries performed in single PCR tubes in the laboratory or on a home workbench. As H3K4 methylation is produced by transcription at promoters and enhancers, our method identifies transcription-coupled accessible regulatory sites.
]]></description>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Henikoff, J. G.</dc:creator>
<dc:date>2020-04-17</dc:date>
<dc:identifier>doi:10.1101/2020.04.15.043083</dc:identifier>
<dc:title><![CDATA[Profiling the epigenome at home]]></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/10.1101/2020.04.16.045443v1?rss=1">
<title>
<![CDATA[
Repairing neural damage in a C. elegans chemosensory circuit using genetically engineered synapses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.16.045443v1?rss=1</link>
<description><![CDATA[
Neuronal loss can considerably diminish neural circuit function, impairing normal behavior by disrupting information flow in the circuit. We reasoned that by rerouting the flow of information in the damaged circuit it may be possible to offset these negative outcomes. We examined this possibility using the well-characterized chemosensory circuit of the nematode worm C. elegans. In this circuit, a main sensory neuron class sends parallel outputs to several interneuron classes. We found that the removal of one of these interneuron classes impairs chemotaxis to attractive odors, revealing a prominent path for information flow in the circuit. To alleviate these deficiencies, we sought to reinforce a remaining neural pathway. We used genetically engineered electrical synapses for this purpose, and observed the successful recovery of chemotaxis performance. However, we were surprised to find that the recovery was largely mediated by inadvertently formed left-right lateral electrical connections within individual neuron classes. Our analysis suggests that these additional electrical synapses help restore circuit function by amplifying weakened neuronal signals in the damaged circuit. These results demonstrate the power of genetically engineered synapses to regulate information flow and signal intensity in damaged neural circuits.
]]></description>
<dc:creator>Rabinowitch, I.</dc:creator>
<dc:creator>Upadhyaya, B.</dc:creator>
<dc:creator>Pant, A.</dc:creator>
<dc:creator>Bai, J.</dc:creator>
<dc:date>2020-04-17</dc:date>
<dc:identifier>doi:10.1101/2020.04.16.045443</dc:identifier>
<dc:title><![CDATA[Repairing neural damage in a C. elegans chemosensory circuit using genetically engineered synapses]]></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/10.1101/2020.04.20.051219v1?rss=1">
<title>
<![CDATA[
Protocol and reagents for pseudotyping lentiviral particles with SARS-CoV-2 Spike protein for neutralization assays 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.20.051219v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 enters cells using its Spike protein, which is also the main target of neutralizing antibodies. Therefore, assays to measure how antibodies and sera affect Spike-mediated viral infection are important for studying immunity. Because SARS-CoV-2 is a biosafety-level-3 virus, one way to simplify such assays is to pseudotype biosafety-level-2 viral particles with Spike. Such pseudotyping has now been described for single-cycle lentiviral, retroviral and VSV particles, but the reagents and protocols are not widely available. Here we detail how to effectively pseudotype lentiviral particles with SARS-CoV-2 Spike and infect 293T cells engineered to express the SARS-CoV-2 receptor, ACE2. We also make all the key experimental reagents available in the BEI Resources repository of ATCC and the NIH. Furthermore, we demonstrate how these pseudotyped lentiviral particles can be used to measure the neutralizing activity of human sera or plasma against SARS-CoV-2 in convenient luciferase-based assays, thereby providing a valuable complement to ELISA-based methods that measure antibody binding rather than neutralization.
]]></description>
<dc:creator>Crawford, K. H. D.</dc:creator>
<dc:creator>Eguia, R.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Malone, K. D.</dc:creator>
<dc:creator>Wolf, C. R.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Tortorici, M. A.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Murphy, M.</dc:creator>
<dc:creator>Pettie, D.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:creator>Balazs, A. B.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-04-20</dc:date>
<dc:identifier>doi:10.1101/2020.04.20.051219</dc:identifier>
<dc:title><![CDATA[Protocol and reagents for pseudotyping lentiviral particles with SARS-CoV-2 Spike protein for neutralization assays]]></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/10.1101/2020.04.22.056283v1?rss=1">
<title>
<![CDATA[
Preliminary support for a 'dry swab, extraction free' protocol for SARS-CoV-2 testing via RT-qPCR 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.22.056283v1?rss=1</link>
<description><![CDATA[
Structured AbstractO_ST_ABSBackgroundC_ST_ABSThe urgent need for massively scaled clinical testing for SARS-CoV-2, along with global shortages of critical reagents and supplies, has necessitated development of streamlined laboratory testing protocols. Conventional nucleic acid testing for SARS-CoV-2 involves collection of a clinical specimen with a nasopharyngeal swab in transport medium, nucleic acid extraction, and quantitative reverse transcription PCR (RT-qPCR) (1). As testing has scaled across the world, the global supply chain has buckled, rendering testing reagents and materials scarce (2). To address shortages, we developed SwabExpress, an end-to-end protocol developed to employ mass produced anterior nares swabs and bypass the requirement for transport media and nucleic acid extraction.

MethodsWe evaluated anterior nares swabs, transported dry and eluted in low-TE buffer as a direct-to-RT-qPCR alternative to extraction-dependent viral transport media. We validated our protocol of using heat treatment for viral activation and added a proteinase K digestion step to reduce amplification interference. We tested this protocol across archived and prospectively collected swab specimens to fine-tune test performance.

ResultsAfter optimization, SwabExpress has a low limit of detection at 2-4 molecules/uL, 100% sensitivity, and 99.4% specificity when compared side-by-side with a traditional RT-qPCR protocol employing extraction. On real-world specimens, SwabExpress outperforms an automated extraction system while simultaneously reducing cost and hands-on time.

ConclusionSwabExpress is a simplified workflow that facilitates scaled testing for COVID-19 without sacrificing test performance. It may serve as a template for the simplification of PCR-based clinical laboratory tests, particularly in times of critical shortages during pandemics.
]]></description>
<dc:creator>Srivatsan, S.</dc:creator>
<dc:creator>Han, P. D.</dc:creator>
<dc:creator>van Raay, K.</dc:creator>
<dc:creator>Wolf, C. R.</dc:creator>
<dc:creator>McCulloch, D. J.</dc:creator>
<dc:creator>Kim, A. E.</dc:creator>
<dc:creator>Brandstetter, E.</dc:creator>
<dc:creator>Martin, B.</dc:creator>
<dc:creator>Gehring, J.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Seattle Flu Study Investigators,</dc:creator>
<dc:creator>Kosuri, S.</dc:creator>
<dc:creator>Konnick, E. Q.</dc:creator>
<dc:creator>Lockwood, C. M.</dc:creator>
<dc:creator>Reider, M. J.</dc:creator>
<dc:creator>Nickerson, D. A.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Starita, L. M.</dc:creator>
<dc:date>2020-04-23</dc:date>
<dc:identifier>doi:10.1101/2020.04.22.056283</dc:identifier>
<dc:title><![CDATA[Preliminary support for a 'dry swab, extraction free' protocol for SARS-CoV-2 testing via RT-qPCR]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.24.060194v1?rss=1">
<title>
<![CDATA[
Using B cell receptor lineage structures to predict affinity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.24.060194v1?rss=1</link>
<description><![CDATA[
We are frequently faced with a large collection of antibodies, and want to select those with highest affinity for their cognate antigen. When developing a first-line therapeutic for a novel pathogen, for instance, we might look for such antibodies in patients that have recovered. There exist effective experimental methods of accomplishing this, such as cell sorting and baiting; how-ever they are time consuming and expensive. Next generation sequencing of B cell receptor (BCR) repertoires offers an additional source of sequences that could be tapped if we had a reliable method of selecting those coding for the best antibodies. In this paper we introduce a method that uses evolutionary information from the family of related sequences that share a naive ancestor to predict the affinity of each resulting antibody for its antigen. When combined with information on the identity of the antigen, this method should provide a source of effective new antibodies. We also introduce a method for a related task: given an antibody of interest and its inferred ancestral lineage, which branches in the tree are likely to harbor key affinity-increasing mutations? These methods are implemented as part of continuing development of the partis BCR inference package, available at https://github.com/psathyrella/partis.

Comments. Please post comments or questions on this paper as new issues at https://git.io/Jvxkn.
]]></description>
<dc:creator>Ralph, D. K.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2020-04-25</dc:date>
<dc:identifier>doi:10.1101/2020.04.24.060194</dc:identifier>
<dc:title><![CDATA[Using B cell receptor lineage structures to predict affinity]]></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/10.1101/2020.04.27.052225v1?rss=1">
<title>
<![CDATA[
Characterising the epidemic spread of Influenza A/H3N2 within a city through phylogenetics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.27.052225v1?rss=1</link>
<description><![CDATA[
Infecting large portions of the global population, seasonal influenza is a major burden on societies around the globe. While the global source sink dynamics of the different seasonal influenza viruses have been studied intensively, its local spread remains less clear. In order to improve our understanding of how influenza is transmitted on a city scale, we collected an extremely densely sampled set of influenza sequences alongside patient metadata. To do so, we sequenced influenza viruses isolated from patients of two different hospitals, as well as private practitioners in Basel, Switzerland during the 2016/2017 influenza season. The genetic sequences reveal that repeated introductions into the city drove the influenza season. We then reconstruct how the effective reproduction number changed over the course of the season. We find trends in transmission dynamics correlated positively with trends in temperature, but not relative humidity nor school holidays. Alongside the genetic sequence data that allows us to see how individual cases are connected, we gathered patient information, such as the age or household status. Zooming into the local transmission outbreaks suggests that the elderly were to a large extent infected within their own transmission network, while school children likely drove the spread within the remaining transmission network. These patterns will be valuable to plan interventions combating the spread of respiratory diseases within cities given that similar patterns are observed for other influenza seasons and cities.

Author summaryAs shown with the current SARS-CoV-2 pandemic, respiratory diseases can quickly spread around the globe. While it can be hugely important to understand how diseases spread around the globe, local spread is most often the main driver of novel infections of respiratory diseases such as SARS-CoV-2 or influenza. We here use genetic sequence data alongside patient information to better understand what the drives the local spread of influenza by looking at the 2016/2017 influenza season in Basel, Switzerland as an example. The genetic sequence data allows us to reconstruct the how the transmission dynamics changed over the course of the season, which we correlate to changes, but not humidity or school holidays. Additionally, the genetic sequence data allows us to see how individual cases are connected. Using patient information, such as age and household status our analyses suggest that the elderly mainly transmit within their own transmission network. Additionally, they suggest that school aged children, but not pre-school aged children are important drivers of the local spread of influenza.
]]></description>
<dc:creator>Müller, N. F.</dc:creator>
<dc:creator>Wüthrich, D.</dc:creator>
<dc:creator>Goldman, N.</dc:creator>
<dc:creator>Sailer, N.</dc:creator>
<dc:creator>Saalfrank, C.</dc:creator>
<dc:creator>Brunner, M.</dc:creator>
<dc:creator>Augustin, N.</dc:creator>
<dc:creator>Seth-Smith, H. M.</dc:creator>
<dc:creator>Hollenstein, Y.</dc:creator>
<dc:creator>Syedbasha, M.</dc:creator>
<dc:creator>Lang, D.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:creator>Dubuis, O.</dc:creator>
<dc:creator>Naegele, M.</dc:creator>
<dc:creator>Buser, A.</dc:creator>
<dc:creator>Nickel, C. H.</dc:creator>
<dc:creator>Ritz, N.</dc:creator>
<dc:creator>Zeller, A.</dc:creator>
<dc:creator>Lang, B. M.</dc:creator>
<dc:creator>Hadfield, J.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Battegay, M.</dc:creator>
<dc:creator>Schneider-Sliwa, R.</dc:creator>
<dc:creator>Egli, A.</dc:creator>
<dc:creator>Stadler, T.</dc:creator>
<dc:date>2020-04-29</dc:date>
<dc:identifier>doi:10.1101/2020.04.27.052225</dc:identifier>
<dc:title><![CDATA[Characterising the epidemic spread of Influenza A/H3N2 within a city through phylogenetics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.27.064774v1?rss=1">
<title>
<![CDATA[
A SARS-CoV-2 vaccine candidate would likely match all currently circulating strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.27.064774v1?rss=1</link>
<description><![CDATA[
The magnitude of the COVID-19 pandemic underscores the urgency for a safe and effective vaccine. Here we analyzed SARS-CoV-2 sequence diversity across 5,700 sequences sampled since December 2019. The Spike protein, which is the target immunogen of most vaccine candidates, showed 93 sites with shared polymorphisms; only one of these mutations was found in more than 1% of currently circulating sequences. The minimal diversity found among SARS-CoV-2 sequences can be explained by drift and bottleneck events as the virus spread away from its original epicenter in Wuhan, China. Importantly, there is little evidence that the virus has adapted to its human host since December 2019. Our findings suggest that a single vaccine should be efficacious against current global strains.

One Sentence SummaryThe limited diversification of SARS-CoV-2 reflects drift and bottleneck events rather than adaptation to humans as the virus spread.
]]></description>
<dc:creator>Dearlove, B. L.</dc:creator>
<dc:creator>Lewitus, E.</dc:creator>
<dc:creator>Bai, H.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Reeves, D. B.</dc:creator>
<dc:creator>Joyce, M. G.</dc:creator>
<dc:creator>Scott, P.</dc:creator>
<dc:creator>Amare, M.</dc:creator>
<dc:creator>Vasan, S.</dc:creator>
<dc:creator>Michael, N. L.</dc:creator>
<dc:creator>Modjarrad, K.</dc:creator>
<dc:creator>Rolland, M.</dc:creator>
<dc:date>2020-04-27</dc:date>
<dc:identifier>doi:10.1101/2020.04.27.064774</dc:identifier>
<dc:title><![CDATA[A SARS-CoV-2 vaccine candidate would likely match all currently circulating strains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.04.076638v1?rss=1">
<title>
<![CDATA[
Tumor-specific changes in Kaposi sarcoma-associated herpesvirus genomes in Ugandan adults with Kaposi sarcoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.04.076638v1?rss=1</link>
<description><![CDATA[
Intra-host evolved tumor virus variants have provided insights into the risk, pathogenesis and treatment responses of associated cancers. However, the intra-host variability of Kaposi sarcoma-associated herpesvirus (KSHV), the etiologic agent of Kaposi sarcoma (KS), has not been explored at the whole viral genome level. An accurate and detailed description of KSHV intra-host diversity in whole KSHV genomes from matching tumors and oral swabs from Ugandan adults with HIV-associated KS was obtained by deep, short read sequencing, using duplex unique molecular identifiers (dUMI) - random double-stranded oligonucleotides that barcode individual DNA molecules before library amplification. This allowed suppression of PCR and sequencing errors down to [~]10-9/base. KSHV genomes were assembled de novo, and identified rearrangements were confirmed by PCR. 131-kb KSHV genome sequences, excluding major repeat regions and averaging 2.3 x 104 reads/base, were successfully obtained from 23 specimens from 9 individuals, including 7 tumor-oral pairs. Sampling more than 100 viral genomes in at least one specimen per individual showed that KSHV genomes were virtually homogeneous within samples and within individuals at the point mutational level. Heterogeneity, if present, was due to point mutations and genomic rearrangements in tumors. In 2 individuals, the same mutations were found in distinct KS tumors. The K8.1 gene was inactivated in tumors from 3 individuals, and all KSHV genomic aberrations retained the region surrounding the first major internal repeat (IR1). These findings suggest that lytic gene alterations may contribute to KS tumorigenesis or persistence.

Author summaryKaposi sarcoma (KS) is a leading cancer in sub-Saharan Africa and in those with HIV co-infection. Infection by Kaposi sarcoma-associated herpesvirus (KSHV) is necessary for KS, yet why only few KSHV infections develop into KS is largely unknown. While strain differences or mutations in other tumor viruses are known to affect the risk and progression of their associated cancers, whether KSHV genetic variation is important to the natural history of KS is unclear. Most studies of KSHV diversity have characterized only [~]4% of its 165-kb genome and may have been impacted by PCR or cloning artifacts. Here, we performed highly sensitive, single-molecule sequencing of whole KSHV genomes in paired KS tumors and oral swabs from 9 individuals with KS. We found that KSHV genomes were virtually identical within individuals, with no evidence of quasispecies formation nor multistrain infection. However, KSHV genome aberrations and inactivating mutations appeared to be a common, tumor-associated phenomenon, with some mutations shared by distinct tumors within an individual. Certain regions of the KSHV genome featured prominently among tumor-associated mutations, suggesting that they are important contributors to the pathogenesis or persistence of KS.
]]></description>
<dc:creator>Santiago, C.</dc:creator>
<dc:creator>Goldman, J. D.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Pankow, A. P.</dc:creator>
<dc:creator>Okuku, F.</dc:creator>
<dc:creator>Schmitt, M. W.</dc:creator>
<dc:creator>Chen, L. H.</dc:creator>
<dc:creator>Hill, C. A.</dc:creator>
<dc:creator>Casper, C.</dc:creator>
<dc:creator>Phipps, W. T.</dc:creator>
<dc:creator>Mullins, J. I.</dc:creator>
<dc:date>2020-05-04</dc:date>
<dc:identifier>doi:10.1101/2020.05.04.076638</dc:identifier>
<dc:title><![CDATA[Tumor-specific changes in Kaposi sarcoma-associated herpesvirus genomes in Ugandan adults with Kaposi sarcoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.08.084954v1?rss=1">
<title>
<![CDATA[
Antibody neutralization of an influenza virus that uses neuraminidase for receptor binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.08.084954v1?rss=1</link>
<description><![CDATA[
Influenza virus infection elicits antibodies against the receptor-binding protein hemagglutinin (HA) and the receptor-cleaving protein neuraminidase (NA). Because HA is essential for viral entry, antibodies targeting HA often potently neutralize the virus in single-cycle infection assays. But antibodies against NA are not potently neutralizing in such assays, since NA is dispensable for single-cycle infection. Here we show that a modified influenza virus that depends on NA for receptor binding is much more sensitive than a virus with receptor-binding HA to neutralization by some anti-NA antibodies. Specifically, virus with a receptor-binding G147R N1 NA and a binding-deficient HA is completely neutralized in single-cycle infections by an antibody that binds near the NA active site. Infection is also substantially inhibited by antibodies that bind NA epitopes distant from the active site. Finally, we demonstrate that this modified virus can be used to efficiently select mutations in NA that escape antibody binding, a task that can be laborious with typical influenza viruses that are not well-neutralized by anti-NA antibodies. Thus, viruses dependent on NA for receptor binding allow for sensitive in vitro detection of antibodies binding near the catalytic site of NA and enable selection of viral escape mutants.
]]></description>
<dc:creator>Gentles, L. E.</dc:creator>
<dc:creator>Wan, H.</dc:creator>
<dc:creator>Eichelberger, M.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-05-09</dc:date>
<dc:identifier>doi:10.1101/2020.05.08.084954</dc:identifier>
<dc:title><![CDATA[Antibody neutralization of an influenza virus that uses neuraminidase for receptor binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.11.089342v1?rss=1">
<title>
<![CDATA[
Phage-DMS: a comprehensive method for fine mapping of antibody epitopes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.11.089342v1?rss=1</link>
<description><![CDATA[
Understanding the antibody response is critical to developing vaccine and antibody-based therapies and has inspired the recent development of new methods to isolate antibodies. However, methods to define the antibody-antigen interactions that determine specificity or allow escape have not kept pace. We developed Phage-DMS, a method which combines two powerful approaches - immunoprecipitation of phage peptide libraries and deep mutational scanning (DMS) - to enable high-throughput fine mapping of antibody epitopes. As an example, we designed sequences encoding all possible amino acid variants of HIV Envelope to create phage display libraries. Using Phage-DMS, we identified sites of escape predicted using other approaches for four well characterized HIV monoclonal antibodies with known linear epitopes. In some cases, the results of Phage-DMS refined the epitope beyond what was determined in previous studies. This method has the potential to rapidly and comprehensively screen many antibodies in a single experiment to define sites essential for binding to antigen.
]]></description>
<dc:creator>Garrett, M. E.</dc:creator>
<dc:creator>Itell, H. L.</dc:creator>
<dc:creator>Crawford, K. H. D.</dc:creator>
<dc:creator>Basom, R.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2020-05-13</dc:date>
<dc:identifier>doi:10.1101/2020.05.11.089342</dc:identifier>
<dc:title><![CDATA[Phage-DMS: a comprehensive method for fine mapping of antibody epitopes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.12.091298v1?rss=1">
<title>
<![CDATA[
Characterization of neutralizing antibodies from a SARS-CoV-2 infected individual 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.12.091298v1?rss=1</link>
<description><![CDATA[
B cells specific for the SARS-CoV-2 S envelope glycoprotein spike were isolated from a COVID-19-infected subject using a stabilized spike-derived ectodomain (S2P) twenty-one days post-infection. Forty-four S2P-specific monoclonal antibodies were generated, three of which bound to the receptor binding domain (RBD). The antibodies were minimally mutated from germline and were derived from different B cell lineages. Only two antibodies displayed neutralizing activity against SARS-CoV-2 pseudo-virus. The most potent antibody bound the RBD in a manner that prevented binding to the ACE2 receptor, while the other bound outside the RBD. Our study indicates that the majority of antibodies against the viral envelope spike that were generated during the first weeks of COVID-19 infection are non-neutralizing and target epitopes outside the RBD. Antibodies that disrupt the SARS-CoV-2 spike-ACE2 interaction can potently neutralize the virus without undergoing extensive maturation. Such antibodies have potential preventive/therapeutic potential and can serve as templates for vaccine-design.

IN BRIEFSARS-CoV-2 infection leads to expansion of diverse B cells clones against the viral spike glycoprotein (S). The antibodies bind S with high affinity despite being minimally mutated. Thus, the development of neutralizing antibody responses by vaccination will require the activation of certain naive B cells without requiring extensive somatic mutation.

HighlightsO_LIAnalysis of early B cell response to SARS-CoV-2 spike protein
C_LIO_LIMost antibodies target non-neutralizing epitopes
C_LIO_LIPotent neutralizing mAb blocks the interaction of the S protein with ACE2
C_LIO_LINeutralizing antibodies are minimally mutated
C_LI
]]></description>
<dc:creator>Seydoux, E.</dc:creator>
<dc:creator>Homad, L. J.</dc:creator>
<dc:creator>MacCamy, A. J.</dc:creator>
<dc:creator>Parks, K. R.</dc:creator>
<dc:creator>Hurlburt, N. K.</dc:creator>
<dc:creator>Jennewein, M. F.</dc:creator>
<dc:creator>Akins, N. R.</dc:creator>
<dc:creator>Stuart, A. B.</dc:creator>
<dc:creator>Wan, Y.-H.</dc:creator>
<dc:creator>Feng, J.</dc:creator>
<dc:creator>Nelson, R.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Cohen, K. W.</dc:creator>
<dc:creator>McElrath, J. M.</dc:creator>
<dc:creator>Englund, J. A.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:date>2020-05-12</dc:date>
<dc:identifier>doi:10.1101/2020.05.12.091298</dc:identifier>
<dc:title><![CDATA[Characterization of neutralizing antibodies from a SARS-CoV-2 infected individual]]></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/10.1101/2020.05.14.084913v1?rss=1">
<title>
<![CDATA[
Integrin αvβ8 on T cells is responsible for suppression of anti-tumor immunity in multiple syngeneic models and is a promising target for tumor immunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.14.084913v1?rss=1</link>
<description><![CDATA[
The v{beta}8 integrin is a key activator of transforming growth factor {beta} (TGF {beta}), which has been shown to inhibit anti-tumor immunity. Previous work has suggested that v{beta}8 on tumor cells could modulate tumor growth and responses to immune checkpoint blockade. We now show that a potent blocking monoclonal antibody against v{beta}8 (ADWA-11) causes growth suppression or complete regression in syngeneic models of squamous cell carcinoma (CCK168), mammary cancer (EMT-6), colon cancer (CT26), and prostate cancer (TRAMPC2), especially when it is combined with other immunomodulators (anti-PD-1, anti-CTLA-4 or 4-1BB) or radiotherapy. v{beta}8 is expressed on tumor cells in some of these models, but tumor cell expression of v{beta}8 is not essential for the beneficial effects of ADWA-11 therapy. v{beta}8 is consistently expressed at highest levels on CD4+CD25+ T cells within tumors, and specific deletion of Itgb8 from T cells is as effective as ADWA-11 in suppressing tumor growth. Treatment with ADWA-11 increases expression of a suite of genes in tumor infiltrating CD8+ T cells that are normally inhibited by TGF{beta} and are involved in tumor cell killing, including Granzyme B and Interferon-{gamma}. These findings solidify v{beta}8 integrin as a promising target for cancer immunotherapy, even for tumors that do not express this integrin.
]]></description>
<dc:creator>Dodagatta-Marri, E.</dc:creator>
<dc:creator>Ma, H.-Y.</dc:creator>
<dc:creator>Liang, B.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Meyer, D. S.</dc:creator>
<dc:creator>Sun, K.-H.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Zirak, B.</dc:creator>
<dc:creator>Rosenblum, M. D.</dc:creator>
<dc:creator>Headley, M.</dc:creator>
<dc:creator>Pinzas, L.</dc:creator>
<dc:creator>Reed, N. I.</dc:creator>
<dc:creator>Del Cid, J. S.</dc:creator>
<dc:creator>Adoumie, M.</dc:creator>
<dc:creator>Hann, B. C.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Giddabasappa, A.</dc:creator>
<dc:creator>Noorbehesht, K.</dc:creator>
<dc:creator>Yang, B.</dc:creator>
<dc:creator>Dal Porto, J.</dc:creator>
<dc:creator>Tsukui, T.</dc:creator>
<dc:creator>Niessen, K.</dc:creator>
<dc:creator>Atakilit, A.</dc:creator>
<dc:creator>Akhurst, R. J.</dc:creator>
<dc:creator>Sheppard, D.</dc:creator>
<dc:date>2020-05-15</dc:date>
<dc:identifier>doi:10.1101/2020.05.14.084913</dc:identifier>
<dc:title><![CDATA[Integrin αvβ8 on T cells is responsible for suppression of anti-tumor immunity in multiple syngeneic models and is a promising target for tumor immunotherapy]]></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/10.1101/2020.05.14.096842v1?rss=1">
<title>
<![CDATA[
dms-view: Interactive visualization tool for deep mutational scanning data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.14.096842v1?rss=1</link>
<description><![CDATA[
Summary and PurposeThe high-throughput technique of deep mutational scanning (DMS) has recently made it possible to experimentally measure the effects of all amino-acid mutations to a protein (Fowler and Fields 2014). Over the past five years, this technique has been used to study dozens of different proteins (Esposito et al. 2019) and answer a variety of research questions. For example, DMS has been used for protein engineering (Wrenbeck, Faber, and Whitehead 2017), understanding the human immune response to viruses (Lee et al. 2019), and interpreting human variation in a clinical setting (Starita et al. 2017; Gelman et al. 2019). Accompanying this proliferation of DMS studies has been the development of software tools (Bloom 2015; Rubin et al. 2017) and databases (Esposito et al. 2019) for data analysis and sharing. However, for many purposes it is important to also integrate and visualize the DMS data in the context of other information, such as the 3-D protein structure or natural sequence-variation data.

Here we describe dms-view (https://dms-view.github.io/), a flexible, web-based, interactive visualization tool for DMS data. dms-view is written in JavaScript and D3, and links site-level and mutation-level DMS data to a 3-D protein structure. The user can interactively select sites of interest to examine the DMS measurements in the context of the protein structure. dms-view tracks the input data and user selections in the URL, making it possible to save specific views of interactively generated visualizations to share with collaborators or to support a published study. Importantly, dms-view takes a flexible input data file so users can easily visualize their own DMS data in the context of protein structures of their choosing, and also incorporate additional information such amino-acid frequencies in natural alignments.
]]></description>
<dc:creator>Hilton, S. K.</dc:creator>
<dc:creator>Huddleston, J. L.</dc:creator>
<dc:creator>Black, A.</dc:creator>
<dc:creator>North, K.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-05-15</dc:date>
<dc:identifier>doi:10.1101/2020.05.14.096842</dc:identifier>
<dc:title><![CDATA[dms-view: Interactive visualization tool for deep mutational scanning data]]></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/10.1101/2020.05.15.098830v1?rss=1">
<title>
<![CDATA[
A system-view of B. pertussis booster vaccine responses in adults primed with whole-cell vs. acellular vaccine in infancy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.15.098830v1?rss=1</link>
<description><![CDATA[
Whole-cell inactivated vaccine against Bordetella pertussis (wP) was substituted in many countries by an acellular subunit vaccine (aP) to reduce side effects. Recent epidemiological studies have shown that aP vaccination in infancy induces less durable immunity than wP vaccination. To determine immunological differences associated with aP vs. wP priming, we performed system-level profiling of the immune response in adults primed with aP vs. wP vaccine in infancy following the Tdap booster vaccination as a surrogate to antigen encounter in vivo. Shared immune responses across cohorts were identified, including an increase of the blood monocyte frequency on day 1, and strong antigen-specific IgG response seven days after boost. Comparing aP and wP primed individuals, we found a subset of aP-primed individuals with higher levels of expression for several genes including CCL3 on day 3 and NFKBIA and ICAM1 on day 7 post immunization. These observations were supported by increased CCL3 concentrations in plasma of aP primed individuals. Contrary to the wP individuals, the CCL3-high aP subset presented boosted PT-specific IgE responses. Furthermore, higher antigen specific IgG4 and IgG3 antibodies against specific vaccine antigens at baseline and post boost of aP individuals was observed, suggesting a long term maintained difference in the IgG subtype response. Overall our findings demonstrate that, while broad immune response patterns to Tdap boost overlap between aP and wP primed individuals, a subset of aP primed individuals present a divergent response. These findings provide candidate targets to study the causes and correlates of waning immunity after aP vaccination.
]]></description>
<dc:creator>da Silva Antunes, R.</dc:creator>
<dc:creator>Pomaznoy, M.</dc:creator>
<dc:creator>Soldevila, F.</dc:creator>
<dc:creator>Babor, M.</dc:creator>
<dc:creator>Bennett, J.</dc:creator>
<dc:creator>Tian, Y.</dc:creator>
<dc:creator>Khalil, N.</dc:creator>
<dc:creator>Qian, Y.</dc:creator>
<dc:creator>Mandava, A.</dc:creator>
<dc:creator>Scheuermann, R. H.</dc:creator>
<dc:creator>Cortese, M.</dc:creator>
<dc:creator>Pulendran, B.</dc:creator>
<dc:creator>Petro, C. D.</dc:creator>
<dc:creator>Gilkes, A.</dc:creator>
<dc:creator>Purcell, L. A.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:creator>Peters, B.</dc:creator>
<dc:date>2020-05-16</dc:date>
<dc:identifier>doi:10.1101/2020.05.15.098830</dc:identifier>
<dc:title><![CDATA[A system-view of B. pertussis booster vaccine responses in adults primed with whole-cell vs. acellular vaccine in infancy]]></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/10.1101/2020.05.18.101709v1?rss=1">
<title>
<![CDATA[
Histone deposition pathways determine the chromatin landscapes of H3.1 and H3.3 K27M oncohistones 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.18.101709v1?rss=1</link>
<description><![CDATA[
Lysine 27-to-methionine (K27M) mutations in the H3.1 or H3.3 histone genes are characteristic of pediatric diffuse midline gliomas (DMGs). These oncohistone mutations dominantly inhibit histone H3K27 trimethylation and silencing, but it is unknown how oncohistone type affects gliomagenesis. We show that the genomic distributions of H3.1 and H3.3 oncohistones in human patient-derived DMG cells are consistent with the DNA replication-coupled deposition of histone H3.1 and the predominant replication-independent deposition of histone H3.3. Although H3K27 trimethylation is reduced for both oncohistone types, H3.3K27M-bearing cells retain some domains, and only H3.1K27M-bearing cells lack H3K27 trimethylation. Neither oncohistone interferes with PRC2 binding. Using Drosophila as a model, we demonstrate that inhibition of H3K27 trimethylation occurs only when H3K27M oncohistones are deposited into chromatin and only when expressed in cycling cells. We propose that oncohistones inhibit the H3K27 methyltransferase as chromatin patterns are being duplicated in proliferating cells, predisposing them to tumorigenesis.
]]></description>
<dc:creator>Sarthy, J. F.</dc:creator>
<dc:creator>Meers, M. P.</dc:creator>
<dc:creator>Janssens, D.</dc:creator>
<dc:creator>Henikoff, J. G.</dc:creator>
<dc:creator>Morris, S.</dc:creator>
<dc:creator>Biery, M.</dc:creator>
<dc:creator>Feldman, H.</dc:creator>
<dc:creator>Paddison, P. J.</dc:creator>
<dc:creator>Vitanza, N.</dc:creator>
<dc:creator>Olson, J. M.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2020-05-19</dc:date>
<dc:identifier>doi:10.1101/2020.05.18.101709</dc:identifier>
<dc:title><![CDATA[Histone deposition pathways determine the chromatin landscapes of H3.1 and H3.3 K27M oncohistones]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.29.123505v1?rss=1">
<title>
<![CDATA[
Multi-ancestry genome-wide gene-sleep interactions identify novel loci for blood pressure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.29.123505v1?rss=1</link>
<description><![CDATA[
Long and short sleep duration are associated with elevated blood pressure (BP), possibly through effects on molecular pathways that influence neuroendocrine and vascular systems. To gain new insights into the genetic basis of sleep-related BP variation, we performed genome-wide gene by short or long sleep duration interaction analyses on four BP traits (systolic BP, diastolic BP, mean arterial pressure, and pulse pressure) across five ancestry groups using 1 degree of freedom (1df) interaction and 2df joint tests. Primary multi-ancestry analyses in 62,969 individuals in stage 1 identified 3 novel loci that were replicated in an additional 59,296 individuals in stage 2, including rs7955964 (FIGNL2/ANKRD33) showing significant 1df interactions with long sleep duration and rs73493041 (SNORA26/C9orf170) and rs10406644 (KCTD15/LSM14A) showing significant 1df interactions with short sleep duration (Pint < 5x10-8). Secondary ancestry-specific two-stage analyses and combined stage 1 and 2 analyses additionally identified 23 novel loci that need external replication, including 3 and 5 loci showing significant 1df interactions with long and short sleep duration, respectively (Pint < 5x10-8). Multiple genes mapped to our 26 novel loci have known functions in sleep-wake regulation, nervous and cardiometabolic systems. We also identified new gene by long sleep interactions near five known BP loci ([&le;]1Mb) including NME7, FAM208A, MKLN1, CEP164, and RGL3/ELAVL3 (Pint < 5x10-8). This study indicates that sleep and primary mechanisms regulating BP may interact to elevate BP level, suggesting novel insights into sleep-related BP regulation.
]]></description>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Noordam, R.</dc:creator>
<dc:creator>Cade, B. E.</dc:creator>
<dc:creator>Schwander, K.</dc:creator>
<dc:creator>Winkler, T. W.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Sung, Y. J.</dc:creator>
<dc:creator>Bentley, A. R.</dc:creator>
<dc:creator>Manning, A. K.</dc:creator>
<dc:creator>Aschard, H.</dc:creator>
<dc:creator>Kilpelainen, T. O.</dc:creator>
<dc:creator>Ilkov, M.</dc:creator>
<dc:creator>Brown, M. R.</dc:creator>
<dc:creator>Horimoto, A. R.</dc:creator>
<dc:creator>Richard, M.</dc:creator>
<dc:creator>Bartz, T. M.</dc:creator>
<dc:creator>Vojinovic, D.</dc:creator>
<dc:creator>Lim, E.</dc:creator>
<dc:creator>Nierenberg, J. L.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Chitrala, K.</dc:creator>
<dc:creator>Rankinen, T.</dc:creator>
<dc:creator>Musani, S. K.</dc:creator>
<dc:creator>Franceschini, N.</dc:creator>
<dc:creator>Rauramaa, R.</dc:creator>
<dc:creator>Alver, M.</dc:creator>
<dc:creator>Zee, P.</dc:creator>
<dc:creator>Harris, S. E.</dc:creator>
<dc:creator>van der Most, P. J.</dc:creator>
<dc:creator>Nolte, I. M.</dc:creator>
<dc:creator>Munroe, P. B.</dc:creator>
<dc:creator>Palmer, N. D.</dc:creator>
<dc:creator>Kuhnel, B.</dc:creator>
<dc:creator>Weiss, S.</dc:creator>
<dc:creator>Wen, W.</dc:creator>
<dc:creator>Hall, K. A.</dc:creator>
<dc:creator>Lyytikainen, L.-P.</dc:creator>
<dc:creator>O'Connell, J. R.</dc:creator>
<dc:creator>Eiriksdottir, G.</dc:creator>
<dc:creator>Launer, L. J.</dc:creator>
<dc:creator>de Vries, P. S.</dc:creator>
<dc:creator>Arking, D.</dc:creator>
<dc:creator>Chen,</dc:creator>
<dc:date>2020-05-31</dc:date>
<dc:identifier>doi:10.1101/2020.05.29.123505</dc:identifier>
<dc:title><![CDATA[Multi-ancestry genome-wide gene-sleep interactions identify novel loci for blood pressure]]></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/10.1101/2020.06.02.129486v1?rss=1">
<title>
<![CDATA[
In vitro and in vivo characterization of a recombinant rhesus cytomegalovirus containing a complete genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.02.129486v1?rss=1</link>
<description><![CDATA[
Cytomegaloviruses (CMVs) are highly adapted to their host species resulting in strict species specificity. Hence, in vivo examination of all aspects of CMV biology employs animal models using host-specific CMVs. Infection of rhesus macaques (RM) with rhesus CMV (RhCMV) has been established as a representative model for infection of humans with HCMV due to the close evolutionary relationships of both host and virus. However, the commonly used 68-1 strain of RhCMV has been passaged in fibroblasts for decades resulting in multiple genomic changes due to tissue culture adaptation that cause reduced viremia in RhCMV-naive animals and limited shedding compared to low passage isolates. Using sequence information from primary RhCMV isolates we constructed a full-length (FL) RhCMV by repairing all presumed mutations in the 68-1 bacterial artificial chromosome (BAC). Inoculation of adult, immunocompetent, RhCMV-naive RM with the reconstituted virus resulted in significant replication in the blood similar to primary isolates of RhCMV and furthermore led to extensive viremia in many tissues at day 14 post infection. In contrast, viral dissemination and viremia was greatly reduced upon deletion of genes also lacking in 68-1. Transcriptome analysis of infected tissues further revealed that chemokine-like genes deleted in 68-1 are among the most highly expressed viral transcripts both in vitro and in vivo consistent with an important immunomodulatory function of the respective proteins. We conclude that FL-RhCMV displays in vitro and in vivo characteristics of a wildtype virus while being amenable to genetic modifications through BAC recombineering techniques.

Author SummaryHuman cytomegalovirus (HCMV) infections are generally asymptomatic in healthy immunocompetent individuals, but HCMV can cause serious disease after congenital infection and in individuals with immunocompromised immune systems. Since HCMV is highly species specific and cannot productively infect immunocompetent laboratory animals, experimental infection of rhesus macaques (RM) with rhesus CMV (RhCMV) has been established as a closely related animal model for HCMV. By employing the unique ability of CMV to elicit robust and lasting cellular immunity, this model has also been instrumental in developing novel CMV-based vaccines against chronic and recurring infections with pathogens such as the human immunodeficiency virus (HIV) and Mycobacterium tuberculosis (Mtb). However, most of this work was conducted with derivatives of the 68-1 strain of RhCMV which has acquired multiple genomic alterations in tissue culture. To model pathogenesis and immunology of clinical HCMV isolates we generated a full-length (FL) RhCMV clone representative of low passage isolates. Infection of RhCMV-naive RM with FL-RhCMV demonstrated viremia and tissue dissemination that was comparable to that of non-clonal low passage isolates. We further demonstrate that FL-RhCMV is strongly attenuated upon deletion of gene regions absent in 68-1 thus demonstrating the usefulness of FL-RhCMV to study RhCMV pathogenesis.
]]></description>
<dc:creator>Taher, H.</dc:creator>
<dc:creator>Mahyari, E.</dc:creator>
<dc:creator>Kreklywich, C.</dc:creator>
<dc:creator>Uebelhoer, L. S.</dc:creator>
<dc:creator>McArdle, M. R.</dc:creator>
<dc:creator>Moström, M. J.</dc:creator>
<dc:creator>Bhusari, A.</dc:creator>
<dc:creator>Nekorchuk, M.</dc:creator>
<dc:creator>Whitmer, T.</dc:creator>
<dc:creator>Scheef, E. A.</dc:creator>
<dc:creator>Sprehe, L. M.</dc:creator>
<dc:creator>Roberts, D.</dc:creator>
<dc:creator>Hughes, C. M.</dc:creator>
<dc:creator>Jackson, K. A.</dc:creator>
<dc:creator>Selseth, A. N.</dc:creator>
<dc:creator>Ventura, A. B.</dc:creator>
<dc:creator>Yue, Y.</dc:creator>
<dc:creator>Schmidt, K. A.</dc:creator>
<dc:creator>Shao, J.</dc:creator>
<dc:creator>Edlefsen, P. T.</dc:creator>
<dc:creator>Smedley, J.</dc:creator>
<dc:creator>Stanton, R. J.</dc:creator>
<dc:creator>Axthelm, M. K.</dc:creator>
<dc:creator>Estes, J. D.</dc:creator>
<dc:creator>Hansen, S. G.</dc:creator>
<dc:creator>Kaur, A.</dc:creator>
<dc:creator>Barry, P. A.</dc:creator>
<dc:creator>Bimber, B. N.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Früh, K.</dc:creator>
<dc:creator>Malouli, D.</dc:creator>
<dc:date>2020-06-02</dc:date>
<dc:identifier>doi:10.1101/2020.06.02.129486</dc:identifier>
<dc:title><![CDATA[In vitro and in vivo characterization of a recombinant rhesus cytomegalovirus containing a complete genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.03.131664v1?rss=1">
<title>
<![CDATA[
Lactoferrin retargets adenoviruses to TLR4 to induce an abortive NLRP3-associated pyroptotic response in human dendritic cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.03.131664v1?rss=1</link>
<description><![CDATA[
Despite decades of investigations, we still poorly grasp the immunogenicity of human adenovirus (HAdV)-based vaccines in humans. In this study, we explored the role of lactoferrin, which belong to the alarmin subset of antimicrobial peptides that provide immediate direct and indirect activity against a range of pathogens following a breach in tissue homeostasis. Lactoferrin is a globular, iron-sequestering, glycoprotein that can increase HAdV infection and maturation of antigen-presenting cells. However, the mechanism by which HAdV-lactoferrin complexes induce maturation is unknown. We show that lactoferrin redirects HAdVs from species B, C, and D to toll-like receptor 4 (TLR4) complexes on human mononuclear phagocyte. TLR4-mediated internalization induces an abortive NLRP3-associated pyroptotic response inducing pro-inflammatory cytokine release and disrupting plasma membrane integrity without cell death. These data impact our understanding of the immunogenicity of HAdV-based vaccines and may provide ways to increase their efficacy.
]]></description>
<dc:creator>Cheneau, C.</dc:creator>
<dc:creator>Eichholz, K.</dc:creator>
<dc:creator>Tran, T. H.</dc:creator>
<dc:creator>Tran, T. T. P.</dc:creator>
<dc:creator>Paris, O.</dc:creator>
<dc:creator>Henriquet, C.</dc:creator>
<dc:creator>Pugniere, M.</dc:creator>
<dc:creator>Kremer, E. J.</dc:creator>
<dc:date>2020-06-03</dc:date>
<dc:identifier>doi:10.1101/2020.06.03.131664</dc:identifier>
<dc:title><![CDATA[Lactoferrin retargets adenoviruses to TLR4 to induce an abortive NLRP3-associated pyroptotic response in human dendritic cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.04.134536v1?rss=1">
<title>
<![CDATA[
Linking T cell receptor sequence to transcriptional profiles with clonotype neighbor graph analysis (CoNGA) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.04.134536v1?rss=1</link>
<description><![CDATA[
Multi-modal single-cell technologies capable of simultaneously assaying gene expression and surface phenotype across large numbers of immune cells have described extensive heterogeneity within these complex populations, in healthy and diseased states. In the case of T cells, these technologies have made it possible to profile clonotype, defined by T cell receptor (TCR) sequence, and phenotype, as reflected in gene expression (GEX) profile, surface protein expression, and peptide:MHC (pMHC) binding, across large and diverse cell populations. These rich, high-dimensional datasets have the potential to reveal new relationships between TCR sequence and T cell phenotype that go beyond identification of features shared by clonally related cells. In order to uncover these connections in an unbiased way, we developed a graph-theoretic approach---clonotype neighbor-graph analysis or "CoNGA"---that identifies correlations between GEX profile and TCR sequence through statistical analysis of a pair of T cell similarity graphs, one in which cells are linked based on gene expression similarity and another in which cells are linked by similarity of TCR sequence. Applying CoNGA across diverse human and mouse T cell datasets uncovered known and novel associations between TCR sequence features and cellular phenotype including the classical invariant T cell subsets; a novel defined population of human blood CD8+ T cells expressing the transcription factors HOBIT and HELIOS, NK-associated receptors, and a biased TCR repertoire, representing a potential previously undescribed lineage of "natural lymphocytes"; a striking association between usage of a specific V-beta gene segment and expression of the EPHB6 gene that is conserved between mouse and human; and TCR sequence determinants of differentiation in developing thymocytes. As the size and scale of single-cell datasets continue to grow, we expect that CoNGA will prove to be a useful tool for deconvolving complex relationships between TCR sequence and cellular state in single-cell applications.
]]></description>
<dc:creator>Schattgen, S.</dc:creator>
<dc:creator>Guion, K.</dc:creator>
<dc:creator>Crawford, J. C.</dc:creator>
<dc:creator>Souquette, A.</dc:creator>
<dc:creator>Martinez Barrio, A.</dc:creator>
<dc:creator>Stubbington, M. J. T.</dc:creator>
<dc:creator>Thomas, P. G.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:date>2020-06-05</dc:date>
<dc:identifier>doi:10.1101/2020.06.04.134536</dc:identifier>
<dc:title><![CDATA[Linking T cell receptor sequence to transcriptional profiles with clonotype neighbor graph analysis (CoNGA)]]></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/10.1101/2020.06.08.140558v1?rss=1">
<title>
<![CDATA[
Increased demand for NAD+ relative to ATP drives aerobic glycolysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.08.140558v1?rss=1</link>
<description><![CDATA[
Aerobic glycolysis, or preferential fermentation of glucose-derived pyruvate to lactate despite available oxygen, is a hallmark of proliferative metabolism that is observed across many organisms and conditions. To better understand why aerobic glycolysis is associated with cell proliferation, we examined the metabolic consequence of activating the pyruvate dehydrogenase complex (PDH) to increase mitochondrial pyruvate oxidation at the expense of fermentation. We find that increasing PDH activity impairs cell proliferation by reducing the nicotinamide adenine dinucleotide cofactor ratio (NAD+/NADH). This change in NAD+/NADH ratio is caused by an increase in mitochondrial membrane potential that impairs mitochondrial electron transport and NAD+ regeneration. Uncoupling mitochondrial respiration from ATP synthesis or increasing ATP hydrolysis restores NAD+/NADH homeostasis and proliferation even when glucose oxidation is increased. These data suggest that when the demand for NAD+ to support oxidation reactions exceeds the demand for ATP consumption in cells, NAD+ regeneration by mitochondrial respiration becomes constrained, promoting fermentation despite available oxygen. This argues that cells engage in aerobic glycolysis when the cellular demand for NAD+ is in excess of the cellular demand for ATP.
]]></description>
<dc:creator>Luengo, A.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Gui, D. Y.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:creator>Zagorulya, M.</dc:creator>
<dc:creator>Do, B. T.</dc:creator>
<dc:creator>Ferriera, R.</dc:creator>
<dc:creator>Naamati, A.</dc:creator>
<dc:creator>Ali, A.</dc:creator>
<dc:creator>Lewis, C. A.</dc:creator>
<dc:creator>Thomas, C. J.</dc:creator>
<dc:creator>Spranger, S.</dc:creator>
<dc:creator>Matheson, N. J.</dc:creator>
<dc:creator>Vander Heiden, M. G.</dc:creator>
<dc:date>2020-06-09</dc:date>
<dc:identifier>doi:10.1101/2020.06.08.140558</dc:identifier>
<dc:title><![CDATA[Increased demand for NAD+ relative to ATP drives aerobic glycolysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.10.144071v1?rss=1">
<title>
<![CDATA[
Helicobacter pylori diversification during chronic infection within a single host generates sub-populations with distinct phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.10.144071v1?rss=1</link>
<description><![CDATA[
Helicobacter pylori chronically infects the stomach of approximately half of the worlds population. Manifestation of clinical diseases associated with H. pylori infection, including cancer, is driven by strain properties and host responses; and as chronic infection persists, both are subject to change. Previous studies have documented frequent and extensive within-host bacterial genetic variation. To define how within-host diversity contributes to phenotypes related to H. pylori pathogenesis, this project leverages a collection of 39 clinical isolates acquired prospectively from a single subject at two time points and from multiple gastric sites. During the six years separating collection of these isolates, this individual, initially harboring a duodenal ulcer, progressed to gastric atrophy and concomitant loss of acid secretion. Whole genome sequence analysis identified 2,232 unique single nucleotide polymorphisms (SNPs) across isolates and a nucleotide substitution rate of 1.3x10-4 substitutions/site/year. Gene ontology analysis identified cell envelope genes among the genes with excess accumulation of nonsynonymous SNPs (nSNPs). A dendrogram based on genetic similarity, clusters isolates from each time point separately. Within time points, there is segregation of subgroups with phenotypic differences in bacterial morphology, ability to induce inflammatory cytokines, and mouse colonization. Higher inflammatory cytokine induction in recent isolates maps to shared polymorphisms in the Cag PAI protein, CagY, while rod morphology in a subgroup of recent isolates mapped to eight mutations in three distinct helical cell shape determining (csd) genes. The presence of subgroups with unique genetic and phenotypic properties suggest complex selective forces and multiple sub-niches within the stomach during chronic infection.

Author SummaryHelicobacter pylori, one of the most common bacterial pathogens colonizing humans, is the main agent responsible for stomach ulcers and cancer. Certain strain types are associated with increased risk of disease, however many factors contributing to disease outcome remain unknown. Prior work has documented genetic diversity among bacterial populations within single individuals, but the impact of this diversity for continued bacterial infection or disease progression remains understudied. In our analysis we examined both genetic and functional features of many stomach isolates from a single individual infected over six years. During these six years the subject shifted from having excess acid production and a duodenal ulcer to lower acid production from gastric atrophy. The 39 isolates form sub-populations based on gene sequence changes that accumulated in the different isolates. In addition to having distinguishing genetic features, these sub-populations also have differences in several bacterial properties, including cell shape, ability to activate immune responses, and colonization in a mouse model of infection. This apparent functional specialization suggests that the bacterial sub-populations may have adapted to distinct sub-niches within the stomach during chronic infection.
]]></description>
<dc:creator>Jackson, L. K.</dc:creator>
<dc:creator>Potter, B.</dc:creator>
<dc:creator>Schneider, S. E.</dc:creator>
<dc:creator>Fitzgibbon, M.</dc:creator>
<dc:creator>Blair, K.</dc:creator>
<dc:creator>Farah, H.</dc:creator>
<dc:creator>Krishna, U.</dc:creator>
<dc:creator>Peek, R.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Salama, N. R.</dc:creator>
<dc:date>2020-06-10</dc:date>
<dc:identifier>doi:10.1101/2020.06.10.144071</dc:identifier>
<dc:title><![CDATA[Helicobacter pylori diversification during chronic infection within a single host generates sub-populations with distinct phenotypes]]></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/10.1101/2020.06.12.148692v1?rss=1">
<title>
<![CDATA[
Structural basis for potent neutralization of SARS-CoV-2 and role of antibody affinity maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.12.148692v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 is a betacoronavirus virus responsible for the COVID-19 pandemic. Here, we determined the X-ray crystal structure of a potent neutralizing monoclonal antibody, CV30, isolated from a patient infected with SARS-CoV-2, in complex with the receptor binding domain (RBD). The structure reveals CV30s epitope overlaps with the human ACE2 receptor binding site thus providing the structural basis for its neutralization by preventing ACE2 binding.
]]></description>
<dc:creator>Hurlburt, N. K.</dc:creator>
<dc:creator>Wan, Y.-H.</dc:creator>
<dc:creator>Stuart, A. B.</dc:creator>
<dc:creator>Feng, J.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:date>2020-06-12</dc:date>
<dc:identifier>doi:10.1101/2020.06.12.148692</dc:identifier>
<dc:title><![CDATA[Structural basis for potent neutralization of SARS-CoV-2 and role of antibody affinity maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.12.149088v1?rss=1">
<title>
<![CDATA[
Mutational resilience of antiviral restriction favors primate TRIM5α in host-virus evolutionary arms races 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.12.149088v1?rss=1</link>
<description><![CDATA[
Host antiviral proteins engage in evolutionary arms races with viruses, in which both sides rapidly evolve at interaction interfaces to gain or evade immune defense. For example, primate TRIM5 uses its rapidly evolving "v1" loop to bind retroviral capsids, and single mutations in this loop can dramatically improve retroviral restriction. However, it is unknown whether such gains of viral restriction are rare, or if they incur loss of pre-existing function against other viruses. Using deep mutational scanning, we comprehensively measured how single mutations in the TRIM5 v1 loop affect restriction of divergent retroviruses. Unexpectedly, we found that the majority of mutations increase antiviral function. Moreover, most random mutations do not disrupt potent viral restriction, even when it is newly acquired via single adaptive substitutions. Our results indicate that TRIM5s adaptive landscape is remarkably broad and mutationally resilient, maximizing its chances of success in evolutionary arms races with retroviruses.
]]></description>
<dc:creator>Tenthorey, J.</dc:creator>
<dc:creator>Young, C.</dc:creator>
<dc:creator>Sodeinde, A.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2020-06-13</dc:date>
<dc:identifier>doi:10.1101/2020.06.12.149088</dc:identifier>
<dc:title><![CDATA[Mutational resilience of antiviral restriction favors primate TRIM5α in host-virus evolutionary arms races]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.15.153478v1?rss=1">
<title>
<![CDATA[
PROTECTIVE ANTIBODIES AGAINST HPIV3 INFECTION 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.15.153478v1?rss=1</link>
<description><![CDATA[
Human parainfluenza virus type III (HPIV3) is a common respiratory pathogen that afflicts children and can be fatal in vulnerable populations, including the immunocompromised. Unfortunately, an effective vaccine or therapeutic is not currently available, resulting in tens of thousands of hospitalizations per year. In an effort to discover a protective antibody against HPIV3, we screened the B cell repertoires from peripheral blood, tonsils, or spleen from healthy children and adults. These analyses yielded five monoclonal antibodies that potently neutralized HPIV3 in vitro. These HPIV3 neutralizing antibodies targeted two non-overlapping epitopes of the HPIV3 F protein, with most targeting the apex. Importantly, prophylactic administration of one of these antibodies, named PI3-E12, resulted in potent protection against HPIV3 infection in cotton rats. Additionally, PI3-E12 could also be used therapeutically to suppress HPIV3 in immunocompromised animals. These results demonstrate the potential clinical utility of PI3-E12 for the prevention or treatment of HPIV3 in both immunocompetent and immunocompromised individuals.
]]></description>
<dc:creator>Boonyaratanakornkit, J.</dc:creator>
<dc:creator>Weidle, C.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Bakthavatsalam, R.</dc:creator>
<dc:creator>Perkins, J.</dc:creator>
<dc:creator>Stewart-Jones, G. B. E.</dc:creator>
<dc:creator>Kwong, P. D.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:date>2020-06-15</dc:date>
<dc:identifier>doi:10.1101/2020.06.15.153478</dc:identifier>
<dc:title><![CDATA[PROTECTIVE ANTIBODIES AGAINST HPIV3 INFECTION]]></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/10.1101/2020.06.16.146803v1?rss=1">
<title>
<![CDATA[
Combining genome-wide studies of breast, prostate, ovarian and endometrial cancers maps cross-cancer susceptibility loci and identifies new genetic associations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.16.146803v1?rss=1</link>
<description><![CDATA[
We report a meta-analysis of breast, prostate, ovarian, and endometrial cancer genome-wide association data (effective sample size: 237,483 cases/317,006 controls). This identified 465 independent lead variants (P<5x10-8) across 192 genomic regions. Four lead variants were >1Mb from previously identified risk loci for the four cancers and an additional 23 lead variant-cancer associations were novel for one of the cancers. Bayesian models supported pleiotropic effects involving at least two cancers at 222/465 lead variants in 118/192 regions. Gene-level association analysis identified 13 shared susceptibility genes (P<2.6x10-6) in 13 regions not previously implicated in any of the four cancers and not uncovered by our variant-level meta-analysis. Several lead variants had opposite effects across cancers, including a cluster of such variants in the TP53 pathway. Fifty-four lead variants were associated with blood cell traits and suggested genetic overlaps with clonal hematopoiesis. Our study highlights the remarkable pervasiveness of pleiotropy across hormone-related cancers, further illuminating their shared genetic and mechanistic origins at variant- and gene-level resolution.
]]></description>
<dc:creator>Kar, S. P.</dc:creator>
<dc:creator>Lindstroem, S.</dc:creator>
<dc:creator>Hung, R.</dc:creator>
<dc:creator>Lawrenson, K.</dc:creator>
<dc:creator>Schmidt, M.</dc:creator>
<dc:creator>O'Mara, T.</dc:creator>
<dc:creator>Glubb, D.</dc:creator>
<dc:creator>Tyrer, J.</dc:creator>
<dc:creator>Schildkraut, J.</dc:creator>
<dc:creator>Chang-Claude, J.</dc:creator>
<dc:creator>Alsulimani, A.</dc:creator>
<dc:creator>Anton, F.</dc:creator>
<dc:creator>Beeghly-Fadiel, A.</dc:creator>
<dc:creator>Bjorge, L.</dc:creator>
<dc:creator>Bodelon, C.</dc:creator>
<dc:creator>Brauch, H.</dc:creator>
<dc:creator>Burghaus, S.</dc:creator>
<dc:creator>Campa, D.</dc:creator>
<dc:creator>Carney, M.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Daly, M.</dc:creator>
<dc:creator>du Bois, A.</dc:creator>
<dc:creator>Ekici, A.</dc:creator>
<dc:creator>Ewing, A.</dc:creator>
<dc:creator>Fasching, P.</dc:creator>
<dc:creator>Flanagan, J. M.</dc:creator>
<dc:creator>Gawelko, J.</dc:creator>
<dc:creator>Giles, G.</dc:creator>
<dc:creator>Hamilton, R.</dc:creator>
<dc:creator>Harris, H.</dc:creator>
<dc:creator>Heitz, F.</dc:creator>
<dc:creator>Hildebrandt, M.</dc:creator>
<dc:creator>Hillemanns, P.</dc:creator>
<dc:creator>Huang, R.-Y.</dc:creator>
<dc:creator>Imaz, L.</dc:creator>
<dc:creator>Irmejs, A.</dc:creator>
<dc:creator>Jakubowska, A.</dc:creator>
<dc:creator>Jensen, A.</dc:creator>
<dc:creator>John, E.</dc:creator>
<dc:creator>Kannisto, P.</dc:creator>
<dc:creator>Karlan, B.</dc:creator>
<dc:creator>Khusnutdinova, E.</dc:creator>
<dc:creator>Kiemeney, L.</dc:creator>
<dc:creator>Kjaer, S.</dc:creator>
<dc:creator>Klapdor, R.</dc:creator>
<dc:creator>Kleiblo</dc:creator>
<dc:date>2020-06-19</dc:date>
<dc:identifier>doi:10.1101/2020.06.16.146803</dc:identifier>
<dc:title><![CDATA[Combining genome-wide studies of breast, prostate, ovarian and endometrial cancers maps cross-cancer susceptibility loci and identifies new genetic associations]]></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/10.1101/2020.06.17.152926v1?rss=1">
<title>
<![CDATA[
Infinity Flow: High-throughput single-cell quantification of 100s of proteins using conventional flow cytometry and machine learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.17.152926v1?rss=1</link>
<description><![CDATA[
Modern immunologic research increasingly requires high-dimensional analyses in order to understand the complex milieu of cell-types that comprise the tissue microenvironments of disease. To achieve this, we developed Infinity Flow combining hundreds of overlapping flow cytometry panels using machine learning to enable the simultaneous analysis of the co-expression patterns of 100s of surface-expressed proteins across millions of individual cells. In this study, we demonstrate that this approach allows the comprehensive analysis of the cellular constituency of the steady-state murine lung and to identify novel cellular heterogeneity in the lungs of melanoma metastasis bearing mice. We show that by using supervised machine learning, Infinity Flow enhances the accuracy and depth of clustering or dimensionality reduction algorithms. Infinity Flow is a highly scalable, low-cost and accessible solution to single cell proteomics in complex tissues.
]]></description>
<dc:creator>Becht, E.</dc:creator>
<dc:creator>Tolstrup, D.</dc:creator>
<dc:creator>Dutertre, C.-A.</dc:creator>
<dc:creator>Ginhoux, F.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Headley, M. B.</dc:creator>
<dc:date>2020-06-19</dc:date>
<dc:identifier>doi:10.1101/2020.06.17.152926</dc:identifier>
<dc:title><![CDATA[Infinity Flow: High-throughput single-cell quantification of 100s of proteins using conventional flow cytometry and machine learning]]></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/10.1101/2020.06.17.157982v1?rss=1">
<title>
<![CDATA[
Deep mutational scanning of SARS-CoV-2 receptor binding domain reveals constraints on folding and ACE2 binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.17.157982v1?rss=1</link>
<description><![CDATA[
The receptor binding domain (RBD) of the SARS-CoV-2 spike glycoprotein mediates viral attachment to ACE2 receptor, and is a major determinant of host range and a dominant target of neutralizing antibodies. Here we experimentally measure how all amino-acid mutations to the RBD affect expression of folded protein and its affinity for ACE2. Most mutations are deleterious for RBD expression and ACE2 binding, and we identify constrained regions on the RBDs surface that may be desirable targets for vaccines and antibody-based therapeutics. But a substantial number of mutations are well tolerated or even enhance ACE2 binding, including at ACE2 interface residues that vary across SARS-related coronaviruses. However, we find no evidence that these ACE2-affinity enhancing mutations have been selected in current SARS-CoV-2 pandemic isolates. We present an interactive visualization and open analysis pipeline to facilitate use of our dataset for vaccine design and functional annotation of mutations observed during viral surveillance.
]]></description>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Hilton, S. K.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Navarro, M. J.</dc:creator>
<dc:creator>Bowen, J. E.</dc:creator>
<dc:creator>Tortorici, M. A.</dc:creator>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-06-17</dc:date>
<dc:identifier>doi:10.1101/2020.06.17.157982</dc:identifier>
<dc:title><![CDATA[Deep mutational scanning of SARS-CoV-2 receptor binding domain reveals constraints on folding and ACE2 binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.21.163550v1?rss=1">
<title>
<![CDATA[
Mathematical modeling explains differential SARS CoV-2 kinetics in lung and nasal passages in remdesivir treated rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.21.163550v1?rss=1</link>
<description><![CDATA[
Remdesivir was recently demonstrated to decrease recovery time in hospitalized patients with SARS-CoV-2 infection. In rhesus macaques, early initiation of remdesivir therapy prevented pneumonia and lowered viral loads in the lung, but viral loads increased in the nasal passages five days after therapy. We developed mathematical models to explain these results. We identified that 1) drug potency is slightly higher in nasal passages than in lungs, 2) viral load decrease in lungs relative to nasal passages during therapy because of infection-dependent generation of refractory cells in the lung, 3) incomplete drug potency in the lung that decreases viral loads even slightly may allow substantially less lung damage, and 4) increases in nasal viral load may occur due to a slight blunting of peak viral load and subsequent decrease of the intensity of the innate immune response, as well as a lack of refractory cells. We also hypothesize that direct inoculation of the trachea in rhesus macaques may not recapitulate natural infection as lung damage occurs more abruptly in this model than in human infection. We demonstrate with sensitivity analysis that a drug with higher potency could completely suppress viral replication and lower viral loads abruptly in the nasal passages as well as the lung.

One Sentence SummaryWe developed a mathematical model to explain why remdesivir has a greater antiviral effect on SARS CoV-2 in lung versus nasal passages in rhesus macaques.
]]></description>
<dc:creator>Goyal, A.</dc:creator>
<dc:creator>Duke, E. R.</dc:creator>
<dc:creator>Cardozo-Ojeda, E. F.</dc:creator>
<dc:creator>Schiffer, J. T.</dc:creator>
<dc:date>2020-06-22</dc:date>
<dc:identifier>doi:10.1101/2020.06.21.163550</dc:identifier>
<dc:title><![CDATA[Mathematical modeling explains differential SARS CoV-2 kinetics in lung and nasal passages in remdesivir treated rhesus macaques]]></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/10.1101/2020.06.22.165225v1?rss=1">
<title>
<![CDATA[
In vivo antiviral host response to SARS-CoV-2 by viral load, sex, and age 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.22.165225v1?rss=1</link>
<description><![CDATA[
Despite limited genomic diversity, SARS-CoV-2 has shown a wide range of clinical manifestations in different patient populations. The mechanisms behind these host differences are still unclear. Here, we examined host response gene expression across infection status, viral load, age, and sex among shotgun RNA-sequencing profiles of nasopharyngeal swabs from 430 individuals with PCR-confirmed SARS-CoV-2 and 54 negative controls. SARS-CoV-2 induced a strong antiviral response with upregulation of antiviral factors such as OAS1-3 and IFIT1-3, and Th1 chemokines CXCL9/10/11, as well as a reduction in transcription of ribosomal proteins. SARS-CoV-2 culture in human airway epithelial cultures replicated the in vivo antiviral host response. Patient-matched longitudinal specimens (mean elapsed time = 6.3 days) demonstrated reduction in interferon-induced transcription, recovery of transcription of ribosomal proteins, and initiation of wound healing and humoral immune responses. Expression of interferon-responsive genes, including ACE2, increased as a function of viral load, while transcripts for B cell-specific proteins and neutrophil chemokines were elevated in patients with lower viral load. Older individuals had reduced expression of Th1 chemokines CXCL9/10/11 and their cognate receptor, CXCR3, as well as CD8A and granzyme B, suggesting deficiencies in trafficking and/or function of cytotoxic T cells and natural killer (NK) cells. Relative to females, males had reduced B and NK cell-specific transcripts and an increase in inhibitors of NF-{kappa}B signaling, possibly inappropriately throttling antiviral responses. Collectively, our data demonstrate that host responses to SARS-CoV-2 are dependent on viral load and infection time course, with observed differences due to age and sex that may contribute to disease severity.
]]></description>
<dc:creator>Lieberman, N. A. P.</dc:creator>
<dc:creator>Peddu, V.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Shrestha, L.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Mears, M. C.</dc:creator>
<dc:creator>Cajimat, M. N.</dc:creator>
<dc:creator>Bente, D. A.</dc:creator>
<dc:creator>Shi, P.-Y.</dc:creator>
<dc:creator>Bovier, F.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Moscona, A.</dc:creator>
<dc:creator>Porotto, M.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:date>2020-06-22</dc:date>
<dc:identifier>doi:10.1101/2020.06.22.165225</dc:identifier>
<dc:title><![CDATA[In vivo antiviral host response to SARS-CoV-2 by viral load, sex, and age]]></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/10.1101/2020.06.23.167718v1?rss=1">
<title>
<![CDATA[
Super LeArner Prediction of NAb Panels (SLAPNAP): A Containerized Tool for Predicting Combination Monoclonal Broadly Neutralizing Antibody Sensitivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.23.167718v1?rss=1</link>
<description><![CDATA[
SummarySingle broadly neutralizing antibody (bnAb) regimens are currently being evaluated in randomized trials for prevention efficacy against HIV-1 infection. Subsequent trials will evaluate combination bnAb regimens (e.g., cocktails, multi-specific antibodies), which demonstrate higher potency and breadth in vitro compared to single bnAbs. Given the large number of potential regimens in the research pipeline, methods for down-selecting these regimens into efficacy trials are of great interest. To aid the down-selection process, we developed Super LeArner Prediction of NAb Panels (SLAPNAP), a software tool for training and evaluating machine learning models that predict in vitro neutralization resistance of HIV Envelope pseudoviruses to a given single or combination bnAb regimen, based on Envelope amino acid sequence features. SLAPNAP also provides measures of variable importance of sequence features. These results can rank bnAb regimens by their potential prevention efficacy and aid assessments of how prevention efficacy depends on sequence features.

Availability and ImplementationSLAPNAP is a freely available docker image that can be downloaded from DockerHub (https://hub.docker.com/r/slapnap/slapnap). Source code and documentation are available at GitHub (respectively, https://github.com/benkeser/slapnap and https://benkeser.github.io/slapnap/).

ContactDavid Benkeser, benkeser@emory.edu
]]></description>
<dc:creator>Benkeser, D.</dc:creator>
<dc:creator>Williamson, B. D.</dc:creator>
<dc:creator>Magaret, C. A.</dc:creator>
<dc:creator>Nizam, S.</dc:creator>
<dc:creator>Gilbert, P. B.</dc:creator>
<dc:date>2020-06-24</dc:date>
<dc:identifier>doi:10.1101/2020.06.23.167718</dc:identifier>
<dc:title><![CDATA[Super LeArner Prediction of NAb Panels (SLAPNAP): A Containerized Tool for Predicting Combination Monoclonal Broadly Neutralizing Antibody Sensitivity]]></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/10.1101/2020.06.23.167759v1?rss=1">
<title>
<![CDATA[
Robust, flexible, and scalable tests for Hardy-Weinberg Equilibrium across diverse ancestries 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.23.167759v1?rss=1</link>
<description><![CDATA[
Traditional Hardy-Weinberg equilibrium (HWE) tests (the {chi}2 test and the exact test) have long been used as a metric for evaluating genotype quality, as technical artifacts leading to incorrect genotype calls often can be identified as deviations from HWE. However, in datasets comprised of individuals from diverse ancestries, HWE can be violated even without genotyping error, complicating the use of HWE testing to assess genotype data quality. In this manuscript, we present the Robust Unified Test for HWE (RUTH) to test for HWE while accounting for population structure and genotype uncertainty, and evaluate the impact of population heterogeneity and genotype uncertainty on the standard HWE tests and alternative methods using simulated and real sequence datasets. Our results demonstrate that ignoring population structure or genotype uncertainty in HWE tests can inflate false positive rates by many orders of magnitude. Our evaluations demonstrate different tradeoffs between false positives and statistical power across the methods, with RUTH consistently amongst the best across all evaluations. RUTH is implemented as a practical and scalable software tool to rapidly perform HWE tests across millions of markers and hundreds of thousands of individuals while supporting standard VCF/BCF formats. RUTH is publicly available at https://www.github.com/statgen/ruth.
]]></description>
<dc:creator>Kwong, A.</dc:creator>
<dc:creator>NHLBI Trans-Omics for Precision Medicine (TOPMed) Consortium,</dc:creator>
<dc:creator>Blackwell, T. W.</dc:creator>
<dc:creator>TOPMed Analysis Working Group,</dc:creator>
<dc:creator>LeFaive, J.</dc:creator>
<dc:creator>de Andrade, M.</dc:creator>
<dc:creator>Barnard, J.</dc:creator>
<dc:creator>Barnes, K. C.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Burchard, E. G.</dc:creator>
<dc:creator>Cade, B. E.</dc:creator>
<dc:creator>Chasman, D.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Conomos, M.</dc:creator>
<dc:creator>Cupples, L. A.</dc:creator>
<dc:creator>Ellinor, P. T.</dc:creator>
<dc:creator>Eng, C.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Irvin, M. R.</dc:creator>
<dc:creator>Kelly, T.</dc:creator>
<dc:creator>Kim, W.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Lubitz, S. A.</dc:creator>
<dc:creator>Mak, A. C. Y.</dc:creator>
<dc:creator>Manichaikul, A. W.</dc:creator>
<dc:creator>Mathias, R. A.</dc:creator>
<dc:creator>Montasser, M. E.</dc:creator>
<dc:creator>Montgomery, C. G.</dc:creator>
<dc:creator>Musani, S.</dc:creator>
<dc:creator>Palmer, N. D.</dc:creator>
<dc:creator>Peloso, G.</dc:creator>
<dc:creator>Qiao, D.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Roden, D. M.</dc:creator>
<dc:creator>Shoemaker, M. B.</dc:creator>
<dc:creator>Smith, J. A</dc:creator>
<dc:date>2020-06-24</dc:date>
<dc:identifier>doi:10.1101/2020.06.23.167759</dc:identifier>
<dc:title><![CDATA[Robust, flexible, and scalable tests for Hardy-Weinberg Equilibrium across diverse ancestries]]></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/10.1101/2020.06.27.175422v1?rss=1">
<title>
<![CDATA[
Identification of cellular context sensitive regulatory variation in mouse genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.27.175422v1?rss=1</link>
<description><![CDATA[
Assessment of the functional consequences of disease-associated sequence variation at non-coding regulatory elements is complicated by their high degree of context sensitivity to both the local chromatin and nuclear environments. Allelic profiling of DNA accessibility across individuals has shown that only a select minority of sequence variation affects transcription factor (TF) occupancy, yet the low sequence diversity in human populations means that no experimental assessment is available for the majority of disease-associated variants. Here we describe high-resolution in vivo maps of allelic DNA accessibility in liver, kidney, lung and B cells from 5 increasingly diverged strains of F1 hybrid mice. The high density of heterozygous sites in these hybrids enables precise quantification of the effect size and cell-type specificity of hundreds of thousands of variants throughout the mouse genome. We show that chromatin-altering variants delineate characteristic sensitivity profiles for hundreds of TF motifs. We develop a compendium of TF-specific sensitivity profiles accounting for genomic context effects. Finally, we link these maps of allelic accessibility to allelic transcript levels in the same samples. This work provides a foundation for quantitative prediction of cell-type specific effects of non-coding variation on TF activity, which will dramatically facilitate both fine-mapping and systems-level analyses of common disease-associated variation in human genomes.
]]></description>
<dc:creator>Maurano, M. T.</dc:creator>
<dc:creator>Halow, J.</dc:creator>
<dc:creator>Byron, R.</dc:creator>
<dc:creator>Groudine, M.</dc:creator>
<dc:creator>Bender, M. A.</dc:creator>
<dc:creator>Stamatoyannopoulos, J. A.</dc:creator>
<dc:date>2020-06-28</dc:date>
<dc:identifier>doi:10.1101/2020.06.27.175422</dc:identifier>
<dc:title><![CDATA[Identification of cellular context sensitive regulatory variation in mouse genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.01.183145v1?rss=1">
<title>
<![CDATA[
A simple and highly efficient method for multi-allelic CRISPR-Cas9 editing in primary cell cultures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.01.183145v1?rss=1</link>
<description><![CDATA[
ABSTRACTBackground CRISPR-Cas9-based technologies have revolutionized experimental manipulation of mammalian genomes. None-the-less, limitations of the delivery and efficacy of these technologies restrict their application in primary cells.Aims To create an optimized protocol for penetrant, reproducible, and fast targeted insertiondeletion mutation (indel) formation in cell cultures derived from primary cells, using patient-derived glioblastoma (GBM) stem-like cells (GSCs) and human neural stem/progenitor cells (NSCs) for proof-of-concept experiments.Methods We employed transient nucleofection of Cas9:sgRNA ribonucleoprotein complexes using chemically synthesized 2’-O-methyl 3’phosphorothioate-modified sgRNAs and purified Cas9 protein. Indel frequency and size distribution were measured via computational deconvolution of Sanger sequencing trace data. Western blotting was used to evaluate protein loss. RNA-seq in edited NSCs was used to assess gene expression changes resulting from knockout of tumor suppressors commonly altered in GBM.Results We found that with this optimized technique, we can routinely achieve &gt;90% indel formation in only 3 days, without the need to create clonal lines for simple loss-of-function experiments. We observed near-total protein loss of target genes in cell pools. Additionally, we found that this approach allows for the creation of targeted genomic deletions. We also demonstrated the utility of this method for quickly creating a series of gene knockouts that allow for the study of oncogenic activities.Conclusion Our data suggest that this relatively simple method can be used for highly efficient and fast gene knockout, as well as for targeted genomic deletions, even in hyperdiploid cells (such as GSCs). This represents an extremely useful tool for the cancer research community when wishing to inactivate not only coding genes, but also non-coding RNAs, UTRs, enhancers, and promoters. This method can be readily applied to diverse cell types by varying the nucleofection conditions.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Hoellerbauer, P.</dc:creator>
<dc:creator>Kufeld, M.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Wu, H.-J.</dc:creator>
<dc:creator>Feldman, H.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:date>2020-07-01</dc:date>
<dc:identifier>doi:10.1101/2020.07.01.183145</dc:identifier>
<dc:title><![CDATA[A simple and highly efficient method for multi-allelic CRISPR-Cas9 editing in primary cell cultures]]></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/10.1101/2020.07.03.187310v1?rss=1">
<title>
<![CDATA[
An integrative oncogene-dependency map identifies unique vulnerabilities of oncogenic RIT1 in lung cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.03.187310v1?rss=1</link>
<description><![CDATA[
ABSTRACTAdvances in precision oncology have transformed cancer therapy from broadly-applied cytotoxic therapy to personalized treatments based on each tumor’s unique molecular alterations. Here we investigate the oncogene-specific dependencies conferred by lung cancer driver variants of KRAS, EGFR, and RIT1. Integrative analysis of genome-wide CRISPR screens in isogenic cell lines identified shared and unique vulnerabilities of each oncogene. The non-identical landscape of dependencies underscores the importance of genotype-guided therapies to maximize tumor responses. Combining genetic screening data with small molecule sensitivity profiling, we identify a unique vulnerability of RIT1-mutant cells to loss of spindle assembly checkpoint regulators. This sensitivity may be related to a novel role of RIT1 in mitosis; we find that oncogenic RIT1M90I alters mitotic timing via weakening of the spindle assembly checkpoint. In addition, we uncovered a specific cooperation of mutant RIT1 with loss of Hippo pathway genes. In human lung cancer, RIT1 mutations and amplifications frequently co-occur with loss of Hippo pathway gene expression. These results provide the first genome-wide atlas of oncogenic RIT1-cooperating factors and genetic dependencies and identify components of the RAS pathway, spindle assembly checkpoint, and Hippo/YAP1 network as candidate therapeutic targets in RIT1-mutant lung cancer.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Vichas, A.</dc:creator>
<dc:creator>Nkinsi, N. T.</dc:creator>
<dc:creator>Riley, A.</dc:creator>
<dc:creator>Parrish, P. C. R.</dc:creator>
<dc:creator>Duke, F.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Watson, J.</dc:creator>
<dc:creator>Rees, M.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:creator>Piccioni, F.</dc:creator>
<dc:creator>Hatch, E. M.</dc:creator>
<dc:creator>Berger, A. H.</dc:creator>
<dc:date>2020-07-04</dc:date>
<dc:identifier>doi:10.1101/2020.07.03.187310</dc:identifier>
<dc:title><![CDATA[An integrative oncogene-dependency map identifies unique vulnerabilities of oncogenic RIT1 in lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.08.192542v1?rss=1">
<title>
<![CDATA[
B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.08.192542v1?rss=1</link>
<description><![CDATA[
Tissue-based T cells increasingly have been shown to be important effectors in the control and prevention of mucosal viral infections - less is known about tissue-based B cells. We demonstrate that B cells and antibody-secreting cells (ASCs) are present in skin biopsies of persons with symptomatic HSV-2 reactivation. CD20+ B cells are observed in inflammatory infiltrates at greatest density at the time of symptomatic reactivation; HSV-2-specific antibodies to HSV-2 surface antigens are also detected. The concentrations of HSV-2-specific antibodies in tissue biopsies vary over the course of HSV-2 reactivation and healing, unlike serum where concentrations remain static over time. B cells and HSV-specific antibody were rarely present in biopsies of unaffected skin. Investigation of serial biopsies over the course of lesion healing suggests that B cells follow a more migratory than resident pattern of infiltration in HSV-affected genital skin, in contrast to T cells. Together, these observations may suggest a functional and distinct role of tissue-based B cells in the local immune response to HSV-2.
]]></description>
<dc:creator>Emily S. Ford</dc:creator>
<dc:creator>Anton M. Sholukh</dc:creator>
<dc:creator>RuthMabel Boytz</dc:creator>
<dc:creator>Savanna S. Carmack</dc:creator>
<dc:creator>Alexis Klock</dc:creator>
<dc:creator>Khamsone Phasouk</dc:creator>
<dc:creator>Jason Shao</dc:creator>
<dc:creator>Raabya Rossenkhan</dc:creator>
<dc:creator>Paul T. Edlefsen</dc:creator>
<dc:creator>Tao Peng</dc:creator>
<dc:creator>Christine Johnston</dc:creator>
<dc:creator>Anna Wald</dc:creator>
<dc:creator>Jia Zhu</dc:creator>
<dc:creator>Lawrence Corey</dc:creator>
<dc:date>2020-07-08</dc:date>
<dc:identifier>doi:10.1101/2020.07.08.192542</dc:identifier>
<dc:title><![CDATA[B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.13.201061v1?rss=1">
<title>
<![CDATA[
Cell Type Aware analysis of RNA-seq data(CARseq) reveals difference and similarities ofthe molecular mechanisms of Schizophrenia andAutism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.13.201061v1?rss=1</link>
<description><![CDATA[
Most tissue samples are composed of different cell types. Differential expression analysis without accounting for cell type composition cannot separate the changes due to cell type composition or cell type-specific expression. We propose a computational framework to address these limitations: Cell Type Aware analysis of RNA-seq (CARseq). CARseq employs a negative binomial distribution that appropriately models the count data from RNA-seq experiments. Simulation studies show that CARseq has substantially higher power than a linear model-based approach and it also provides more accurate estimate of the rankings of differentially expressed genes. We have applied CARseq to compare gene expression of schizophrenia/autism subjects versus controls, and identified the cell types underlying the difference and similarities of these two neuron-developmental diseases. Our results are consistent with the results from differential expression analysis using single cell RNA-seq data.
]]></description>
<dc:creator>Jin, C.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Lin, D.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2020-07-14</dc:date>
<dc:identifier>doi:10.1101/2020.07.13.201061</dc:identifier>
<dc:title><![CDATA[Cell Type Aware analysis of RNA-seq data(CARseq) reveals difference and similarities ofthe molecular mechanisms of Schizophrenia andAutism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.15.204172v1?rss=1">
<title>
<![CDATA[
Bystander CD4+ T cells infiltrate human tumors and are phenotypically distinct 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.15.204172v1?rss=1</link>
<description><![CDATA[
Tumor-specific T cells likely underpin effective immune checkpoint-blockade therapies. Yet, most studies focus on Treg cells and CD8+ tumor-infiltrating lymphocytes (TILs). Here we study CD4+ TILs in human lung and colorectal cancers and observe that non-Treg CD4+ TILs average more than 70% of total CD4+ TILs in both cancer types. Leveraging high dimensional analyses including mass cytometry and single-cell sequencing, we reveal that CD4+ TILs are heterogeneous at both gene and protein levels, within each tumor and across patients. Consistently, we find different subsets of CD4+ TILs showing characteristics of effectors, tissue resident memory (Trm) or exhausted cells (expressing PD-1, CTLA-4 and CD39). In both cancer types, the frequencies of CD39- non-Treg CD4+ TILs strongly correlate with frequencies of CD39- CD8+ TILs, which we and others have previously shown to be enriched for cells specific for cancer-unrelated antigens (bystanders). Ex-vivo, we demonstrate that CD39- CD4+ TILs can be specific for cancer unrelated antigens, such as HCMV epitopes. Overall, our findings highlight that CD4+ TILs cells are not necessarily tumor-specific and suggest measuring CD39 expression as a straightforward way to quantify or isolate bystander CD4+ T cells.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=158 SRC="FIGDIR/small/204172v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>SIMONI, y.</dc:creator>
<dc:creator>LI, S.</dc:creator>
<dc:creator>ZHUANG, S.</dc:creator>
<dc:creator>HEIT, A.</dc:creator>
<dc:creator>KOO, S.-L.</dc:creator>
<dc:creator>CHOW, I.-T.</dc:creator>
<dc:creator>KWOK, W. W.</dc:creator>
<dc:creator>TAN, I. B.</dc:creator>
<dc:creator>TAN, D. S. W.</dc:creator>
<dc:creator>NEWELL, E. W.</dc:creator>
<dc:date>2020-07-16</dc:date>
<dc:identifier>doi:10.1101/2020.07.15.204172</dc:identifier>
<dc:title><![CDATA[Bystander CD4+ T cells infiltrate human tumors and are phenotypically distinct]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.03.234559v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 ORF6 disrupts nucleocytoplasmic transport through interactions with Rae1 and Nup98 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.03.234559v1?rss=1</link>
<description><![CDATA[
RNA viruses that replicate in the cytoplasm often disrupt nucleocytoplasmic transport to preferentially translate their own transcripts and prevent host antiviral responses. The Sarbecovirus accessory protein ORF6 has previously been shown to be the major inhibitor of interferon production in both SARS-CoV and SARS-CoV-2. SARS-CoV-2 ORF6 was recently shown to co-purify with the host mRNA export factors Rae1 and Nup98. Here, we demonstrate SARS-CoV-2 ORF6 strongly represses protein expression of co-transfected reporter constructs and imprisons host mRNA in the nucleus, which is associated with its ability to co-purify with Rae1 and Nup98. These protein-protein interactions map to the C-terminus of ORF6 and can be abolished by a single amino acid mutation in Met58. Overexpression of Rae1 restores reporter expression in the presence of SARS-CoV-2 ORF6. We further identify an ORF6 mutant containing a 9-amino acid deletion, ORF6 {Delta}22-30, in multiple SARS-CoV-2 clinical isolates that can still downregulate the expression of a co-transfected reporter and interact with Rae1 and Nup98. SARS-CoV ORF6 also interacts with Rae1 and Nup98. However, SARS-CoV-2 ORF6 more strongly co-purifies with Rae1 and Nup98 and results in significantly reduced expression of reporter proteins compared to SARS-CoV ORF6, a potential mechanism for the delayed symptom onset and pre-symptomatic transmission uniquely associated with the SARS-CoV-2 pandemic.

ImportanceSARS-CoV-2, the causative agent of COVID-19, is an RNA virus with a large genome that encodes accessory proteins. While these accessory proteins are not required for growth in vitro, they can contribute to the pathogenicity of the virus. One of SARS-CoV-2s accessory proteins, ORF6, was recently shown to co-purify with two host proteins, Rae1 and Nup98, involved in mRNA nuclear export. We demonstrate SARS-CoV-2 ORF6 interaction with these proteins is associated with reduced expression of a reporter protein and accumulation of poly-A mRNA within the nucleus. SARS-CoV ORF6 also shows the same interactions with Rae1 and Nup98. However, SARS-CoV-2 ORF6 more strongly represses reporter expression and co-purifies with Rae1 and Nup98 compared to SARS-CoV ORF6. The ability of SARS-CoV-2 ORF6 to more strongly disrupt nucleocytoplasmic transport than SARS-CoV ORF6 may partially explain critical differences in clinical presentation between the two viruses.
]]></description>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Lieberman, N. A. P.</dc:creator>
<dc:creator>Phung, Q.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Loprieno, M. A.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Shrestha, L.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:date>2020-08-03</dc:date>
<dc:identifier>doi:10.1101/2020.08.03.234559</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 ORF6 disrupts nucleocytoplasmic transport through interactions with Rae1 and Nup98]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.03.235325v1?rss=1">
<title>
<![CDATA[
chTOG is a conserved mitotic error correction factor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.03.235325v1?rss=1</link>
<description><![CDATA[
Accurate chromosome segregation requires kinetochores on duplicated chromatids to biorient by attaching to dynamic microtubules from opposite spindle poles, which exerts forces to bring kinetochores under tension. However, kinetochores initially bind to MTs indiscriminately, resulting in errors that must be corrected. While the Aurora B protein kinase destabilizes low-tension attachments by phosphorylating kinetochores, low-tension attachments are intrinsically less stable than those under higher tension in vitro independent of Aurora activity. Intrinsic tensionsensitive behavior requires the microtubule regulator Stu2 (budding yeast Dis1/XMAP215 ortholog), which we demonstrate here is likely a conserved function for the TOG protein family. The human TOG protein, chTOG, localizes to kinetochores independent of microtubules by interacting with Hec1. We identify a chTOG mutant that regulates microtubule dynamics but accumulates erroneous kinetochore-microtubule attachments that Aurora B fails to destabilize. Thus, TOG proteins confer a unique, intrinsic error correction activity to kinetochores that ensures accurate chromosome segregation.
]]></description>
<dc:creator>Herman, J. A.</dc:creator>
<dc:creator>Miller, M. P.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2020-08-04</dc:date>
<dc:identifier>doi:10.1101/2020.08.03.235325</dc:identifier>
<dc:title><![CDATA[chTOG is a conserved mitotic error correction factor]]></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/10.1101/2020.08.07.242305v1?rss=1">
<title>
<![CDATA[
A Detailed View of KIR Haplotype Structures and Gene Families as Provided by a New Motif-based Multiple Sequence Alignment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.07.242305v1?rss=1</link>
<description><![CDATA[
Human chromosome 19q13.4 contains genes encoding killer-cell immunoglobulin-like receptors (KIR). Reported haplotype lengths range from 67 to 269 kilobases and contain 4 to 18 genes. The region has certain properties such as single nucleotide variation, structural variation, homology, and repetitive elements that make it hard to align accurately beyond single gene alleles. To the best of our knowledge, a multiple sequence alignment of KIR haplotypes has never been published or presented. Such an alignment would be useful to precisely define KIR haplotypes and loci, provide context for assigning alleles (especially fusion alleles) to genes, infer evolutionary history, impute alleles, interpret and predict co-expression, and generate markers. In order to extend the framework of KIR haplotype sequences in the human genome reference, 27 new sequences were generated including 24 haplotypes from 12 individuals of African American ancestry that were selected for genotypic diversity and novelty to the reference, to bring the total to 68 full length genomic KIR haplotype sequences. We leveraged these data and tools from our long-read KIR haplotype assembly algorithm to define and align KIR haplotypes at <5 kb resolution on average. We then used a standard alignment algorithm to refine that alignment down to single base resolution. This processing demonstrated that the high-level alignment recapitulates human-curated annotation of the human haplotypes as well as a chimpanzee haplotype. Further, assignments and alignments of gene alleles were consistent with their human curation in haplotype and allele databases. These results define KIR haplotypes as 14 loci containing 9 genes. The multiple sequence alignments have been applied in two software packages as probes to capture and annotate KIR haplotypes and as markers to genotype KIR from WGS.
]]></description>
<dc:creator>Roe, D.</dc:creator>
<dc:creator>Vierra-Green, C.</dc:creator>
<dc:creator>Pyo, C.-W.</dc:creator>
<dc:creator>Geraghty, D. E.</dc:creator>
<dc:creator>Spellman, S. R.</dc:creator>
<dc:creator>Maiers, M.</dc:creator>
<dc:creator>Kuang, R.</dc:creator>
<dc:date>2020-08-09</dc:date>
<dc:identifier>doi:10.1101/2020.08.07.242305</dc:identifier>
<dc:title><![CDATA[A Detailed View of KIR Haplotype Structures and Gene Families as Provided by a New Motif-based Multiple Sequence Alignment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.12.248823v1?rss=1">
<title>
<![CDATA[
Attenuated influenza virions expressing the SARS- CoV-2 receptor-binding domain induce neutralizing antibodies in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.12.248823v1?rss=1</link>
<description><![CDATA[
An effective vaccine is essential to controlling the spread of SARS-CoV-2 virus. Here, we describe an influenza-virus-based vaccine for SARS-CoV-2. We incorporated a membrane-anchored form of the SARS-CoV-2 Spike receptor binding domain (RBD) in place of the neuraminidase (NA) coding sequence in an influenza virus also possessing a mutation that reduces the affinity of hemagglutinin for its sialic acid receptor. The resulting {Delta}NA(RBD)-Flu virus can be generated by reverse genetics and grown to high titers in cell culture. A single-dose intranasal inoculation of mice with {Delta}NA(RBD)-Flu elicits serum neutralizing antibody titers against SAR-CoV-2 comparable to those observed in humans following natural infection ([~]1:200). Furthermore, {Delta}NA(RBD)-Flu itself causes no apparent disease in mice. It might be possible to produce a vaccine similar to {Delta}NA(RBD)-Flu at scale by leveraging existing platforms for production of influenza vaccines.
]]></description>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Gentles, L. E.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-08-13</dc:date>
<dc:identifier>doi:10.1101/2020.08.12.248823</dc:identifier>
<dc:title><![CDATA[Attenuated influenza virions expressing the SARS- CoV-2 receptor-binding domain induce neutralizing antibodies in mice]]></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/10.1101/2020.08.14.239871v1?rss=1">
<title>
<![CDATA[
A combined proteomics and Mendelian randomization approach to investigate the effects of aspirin-targeted proteins on colorectal cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.14.239871v1?rss=1</link>
<description><![CDATA[
BackgroundEvidence for aspirins chemopreventative properties on colorectal cancer (CRC) is substantial, but its mechanism of action is not well-understood. We combined a proteomic approach with Mendelian randomization (MR) to identify possible new aspirin targets that decrease CRC risk.

MethodsHuman colorectal adenoma cells (RG/C2) were treated with aspirin (24 hours) and a stable isotope labelling with amino acids in cell culture (SILAC) based proteomics approach identified altered protein expression. Protein quantitative trait loci (pQTLs) from INTERVAL (N=3,301) and expression QTLs (eQTLs) from the eQTLGen Consortium (N=31,684) were used as genetic proxies for protein and mRNA expression levels. Two-sample MR of mRNA/protein expression on CRC risk was performed using eQTL/pQTL data combined with CRC genetic summary data from the Colon Cancer Family Registry (CCFR), Colorectal Transdisciplinary (CORECT), Genetics and Epidemiology of Colorectal Cancer (GECCO) consortia and UK Biobank (55,168 cases and 65,160 controls).

ResultsAltered expression was detected for 125/5886 proteins. Of these, aspirin decreased MCM6, RRM2 and ARFIP2 expression and MR analysis showed that a standard deviation increase in mRNA/protein expression was associated with increased CRC risk (OR:1.08, 95% CI:1.03-1.13, OR:3.33, 95% CI:2.46-4.50 and OR:1.15, 95% CI:1.02-1.29, respectively).

ConclusionMCM6 and RRM2 are involved in DNA repair whereby reduced expression may lead to increased DNA aberrations and ultimately cancer cell death, whereas ARFIP2 is involved in actin cytoskeletal regulation indicating a possible role in aspirins reduction of metastasis.

ImpactOur approach has shown how laboratory experiments and population-based approaches can combine to identify aspirin-targeted proteins possibly affecting CRC risk.
]]></description>
<dc:creator>Nounu, A.</dc:creator>
<dc:creator>Greenhough, A.</dc:creator>
<dc:creator>Heesom, K. J.</dc:creator>
<dc:creator>Richmond, R. C.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Weinstein, S. J.</dc:creator>
<dc:creator>Albanes, D.</dc:creator>
<dc:creator>Baron, J. A.</dc:creator>
<dc:creator>Hopper, J. L.</dc:creator>
<dc:creator>Figueiredo, J. C.</dc:creator>
<dc:creator>Newcomb, P. A.</dc:creator>
<dc:creator>Lindor, N. M.</dc:creator>
<dc:creator>Casey, G.</dc:creator>
<dc:creator>Platz, E. A.</dc:creator>
<dc:creator>Le Marchand, L.</dc:creator>
<dc:creator>Ulrich, C. M.</dc:creator>
<dc:creator>Li, C. I.</dc:creator>
<dc:creator>van Duijnhoven, F. J.</dc:creator>
<dc:creator>Gsur, A.</dc:creator>
<dc:creator>Campbell, P. T.</dc:creator>
<dc:creator>Moreno, V.</dc:creator>
<dc:creator>Vodicka, P.</dc:creator>
<dc:creator>Vodickova, L.</dc:creator>
<dc:creator>Brenner, H.</dc:creator>
<dc:creator>Chang-Claude, J.</dc:creator>
<dc:creator>Hoffmeister, M.</dc:creator>
<dc:creator>Sakoda, L. C.</dc:creator>
<dc:creator>Slattery, M. L.</dc:creator>
<dc:creator>Schoen, R. E.</dc:creator>
<dc:creator>Gunter, M. J.</dc:creator>
<dc:creator>Castellvi-Bel, S.</dc:creator>
<dc:creator>Rok Kim, H.</dc:creator>
<dc:creator>Kweon, S.-S.</dc:creator>
<dc:creator>Chan, A. T.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Zheng, W.</dc:creator>
<dc:creator>Bishop, D. T.</dc:creator>
<dc:creator>Buchanan, D. D.</dc:creator>
<dc:creator>Giles, G. G.</dc:creator>
<dc:creator>Gruber, S. B.</dc:creator>
<dc:creator>Rennert, G.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2020-08-14</dc:date>
<dc:identifier>doi:10.1101/2020.08.14.239871</dc:identifier>
<dc:title><![CDATA[A combined proteomics and Mendelian randomization approach to investigate the effects of aspirin-targeted proteins on colorectal cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.14.248864v1?rss=1">
<title>
<![CDATA[
The origins and consequences of UPF1 variants in pancreatic adenosquamous carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.14.248864v1?rss=1</link>
<description><![CDATA[
Pancreatic adenosquamous carcinoma (PASC) is a rare and aggressive subtype of pancreatic cancer whose mutational origins are poorly understood. An early study reported somatic mutations in UPF1, which encodes a core component of the nonsense-mediated mRNA decay (NMD) pathway, as a common signature of PASC, but subsequent studies did not observe these lesions in other PASC cohorts. The corresponding controversy about whether UPF1 mutations are important contributors to PASC has been exacerbated by a paucity of functional studies of these lesions. Here, we systematically assessed the potential roles of UPF1 mutations in PASC. We modeled two reported UPF1 mutations to find no consistent effects on pancreatic cancer growth, acquisition of adenosquamous features, UPF1 splicing, UPF1 protein levels, or NMD efficiency. We subsequently discovered that ~40% of UPF1 mutations reportedly present in PASCs are identical to standing genetic variation in the human population, suggesting that they are likely non-pathogenic inherited variation rather than pathogenic mutations. Our data suggest that UPF1 is not a common functional driver of PASC and motivate further attempts to identify unique genetic features defining these malignancies.
]]></description>
<dc:creator>Polaski, J. T.</dc:creator>
<dc:creator>Udy, D. B.</dc:creator>
<dc:creator>Escobar-Hoyos, L. F.</dc:creator>
<dc:creator>Askan, G.</dc:creator>
<dc:creator>Leach, S. D.</dc:creator>
<dc:creator>Ventura, A.</dc:creator>
<dc:creator>Kannan, R.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:date>2020-08-14</dc:date>
<dc:identifier>doi:10.1101/2020.08.14.248864</dc:identifier>
<dc:title><![CDATA[The origins and consequences of UPF1 variants in pancreatic adenosquamous carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.15.250589v1?rss=1">
<title>
<![CDATA[
Utilizing Computational Machine Learning Tools to Understand Immunogenic Breadth in the Context of a CD8 T-Cell Mediated HIV Response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.15.250589v1?rss=1</link>
<description><![CDATA[
Predictive models are becoming more and more commonplace as tools for candidate antigen discovery to meet the challenges of enabling epitope mapping of cohorts with diverse HLA properties. Here we build on the concept of using two key parameters, diversity metric of the HLA profile of individuals within a population and consideration of sequence diversity in the context of an individuals CD8 T-cell immune repertoire to assess the HIV proteome for defined regions of immunogenicity. Using this approach, Analysis of HLA adaptation and functional immunogenicity data enabled the identification of regions within the proteome that offer significant conservation, HLA recognition within a population, low prevalence of HLA adaptation and demonstrated immunogenicity. We believe this unique and novel approach to vaccine design that, in combination with in vitro functional assays, offers a bespoke pipeline for expedited and rational CD8 T-cell vaccine design for HIV and potentially other pathogens with the potential for both global and local coverage.
]]></description>
<dc:creator>McGowan, E.</dc:creator>
<dc:creator>Rosenthal, R.</dc:creator>
<dc:creator>Fiore-Gartland, A.</dc:creator>
<dc:creator>Macharia, G.</dc:creator>
<dc:creator>Balinda, S.</dc:creator>
<dc:creator>Kapaata, A.</dc:creator>
<dc:creator>Muok, E.</dc:creator>
<dc:creator>Umviligihozo, G.</dc:creator>
<dc:creator>Dalel, J.</dc:creator>
<dc:creator>Streatfield, C.</dc:creator>
<dc:creator>Coutinho, H.</dc:creator>
<dc:creator>Monaco, D.</dc:creator>
<dc:creator>Morrison, D.</dc:creator>
<dc:creator>Yue, L.</dc:creator>
<dc:creator>Hunter, E.</dc:creator>
<dc:creator>Nielsen, M.</dc:creator>
<dc:creator>Gilmour, J.</dc:creator>
<dc:creator>Hare, J.</dc:creator>
<dc:date>2020-08-15</dc:date>
<dc:identifier>doi:10.1101/2020.08.15.250589</dc:identifier>
<dc:title><![CDATA[Utilizing Computational Machine Learning Tools to Understand Immunogenic Breadth in the Context of a CD8 T-Cell Mediated HIV Response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.17.253146v1?rss=1">
<title>
<![CDATA[
Cooperative Binding of Transcription Factors is a Hallmark of Active Enhancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.17.253146v1?rss=1</link>
<description><![CDATA[
Enhancers harbor binding motifs that recruit transcription factors (TFs) for gene activation. While cooperative binding of TFs at enhancers is known to be critical for transcriptional activation of a handful of developmental enhancers, the extent TF cooperativity genome-wide is unknown. Here, we couple high-resolution nuclease footprinting with single-molecule methylation profiling to characterize TF cooperativity at active enhancers in the Drosophila genome. Enrichment of short MNase-protected DNA segments indicates that the majority of enhancers harbor two or more TF binding sites, and we uncover protected fragments that correspond to co-bound sites in thousands of enhancers. We integrate MNase-seq, methylation accessibility profiling, and CUT&RUN chromatin profiling as a comprehensive strategy to characterize co-binding of the Trithorax-like (TRL) DNA binding protein and multiple other TFs and identify states where an enhancer is bound by no TF, by either single factor, by multiple factors, or where binding sites are occluded by nucleosomes. From the analysis of co-binding, we find that cooperativity dominates TF binding in vivo at a majority of active enhancers. TF cooperativity can occur without apparent protein-protein interactions and provides a mechanism to effectively clear nucleosomes and promote enhancer function.
]]></description>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Ramachandran, S.</dc:creator>
<dc:date>2020-08-18</dc:date>
<dc:identifier>doi:10.1101/2020.08.17.253146</dc:identifier>
<dc:title><![CDATA[Cooperative Binding of Transcription Factors is a Hallmark of Active Enhancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.21.260935v1?rss=1">
<title>
<![CDATA[
A CAR RNA FISH assay reveals functional and spatial heterogeneity of chimeric antigen receptor T cells in tissue 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.21.260935v1?rss=1</link>
<description><![CDATA[
Chimeric antigen receptor (CAR) T cells are engineered cells used in cancer therapy and are studied to treat infectious diseases. Trafficking and persistence of CAR T cells is an important requirement for efficacy to target cancer. Here, we describe a CAR RNA FISH histo-cytometry platform combined with a random reaction seed image analysis algorithm to quantitate spatial distribution and in vivo functional activity of a CAR T cell population at a single cell resolution for preclinical models. In situ, CAR T cell exhibited a heterogenous effector gene expression and this was related to the distance from tumor cells, allowing a quantitative assessment of the potential in vivo effectiveness. The platform offers the potential to study immune functions of genetically engineered cells in situ with their target cells in tissues with high statistical power and thus, can serve as an important tool for preclinical assessment of CAR T cell effectiveness.

Brief summaryWe developed an imaging platform and analysis pipeline to study large populations of engineered cells at a single cell level in situ.

One Sentence SummaryWe developed a CAR RNA FISH assay to study chimeric antigen receptor T cell trafficking and function in mouse tissue.
]]></description>
<dc:creator>Eichholz, K.</dc:creator>
<dc:creator>Li, A. Z.</dc:creator>
<dc:creator>Diem, K.</dc:creator>
<dc:creator>Tareen, S. U.</dc:creator>
<dc:creator>Jensen, M.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Corey, L.</dc:creator>
<dc:date>2020-08-24</dc:date>
<dc:identifier>doi:10.1101/2020.08.21.260935</dc:identifier>
<dc:title><![CDATA[A CAR RNA FISH assay reveals functional and spatial heterogeneity of chimeric antigen receptor T cells in tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.31.273631v1?rss=1">
<title>
<![CDATA[
A JAK/STAT-Mediated Inflammatory Signaling Cascade Drives Oncogenesis In AF10-Rearranged AML 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.31.273631v1?rss=1</link>
<description><![CDATA[
Leukemias bearing fusions of the AF10/MLLT10 gene are associated with poor prognosis, and therapies targeting these fusion proteins are lacking. To understand mechanisms underlying AF10 fusion-mediated leukemogenesis, we generated inducible mouse models of AML driven by the most common AF10 fusion proteins, PICALM/CALM-AF10 and KMT2A/MLL-AF10, and performed comprehensive characterization of the disease using transcriptomic, epigenomic, proteomic, and functional genomic approaches. Our studies provide a comprehensive map of gene networks and protein interactors associated with key AF10 fusions involved in leukemia. Specifically, we report that AF10 fusions activate a cascade of JAK/STAT-mediated inflammatory signaling through direct recruitment of JAK1 kinase. Inhibition of the JAK/STAT signaling by genetic Jak1 deletion or through pharmacological JAK/STAT inhibition elicited potent anti-oncogenic effects in mouse and human models of AF10 fusion AML. Collectively, our study identifies JAK1 as a tractable therapeutic target in AF10-rearranged leukemias.

STATEMENT OF SIGNIFICANCEGene fusions of AF10/MLLT10 are recurrent in acute myeloid and lymphoid leukemia and are associated with extremely poor survival outcomes. We show that the JAK1 kinase is required for activation of the AF10 fusion oncotranscriptome and for leukemogenesis. Since a number of JAK/STAT pathways inhibitors are in clinical development or approved for use, our studies may help develop a therapeutic strategy for AF10-rearranged leukemias.
]]></description>
<dc:creator>Chen, B.</dc:creator>
<dc:creator>Deshpande, A.</dc:creator>
<dc:creator>Barbosa, K. O.</dc:creator>
<dc:creator>Kleppe, M.</dc:creator>
<dc:creator>Lei, X.</dc:creator>
<dc:creator>Yeddula, N.</dc:creator>
<dc:creator>Rosa Campos, A.</dc:creator>
<dc:creator>Wechsler-Reya, R.</dc:creator>
<dc:creator>Bagchi, A.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Eaves, C. J.</dc:creator>
<dc:creator>Jeremias, I.</dc:creator>
<dc:creator>Haferlach, T.</dc:creator>
<dc:creator>Frank, D.</dc:creator>
<dc:creator>Ronai, Z.</dc:creator>
<dc:creator>Chanda, S.</dc:creator>
<dc:creator>Armstrong, S.</dc:creator>
<dc:creator>Adams, P.</dc:creator>
<dc:creator>Levine, R.</dc:creator>
<dc:creator>Deshpande, A.</dc:creator>
<dc:date>2020-08-31</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.273631</dc:identifier>
<dc:title><![CDATA[A JAK/STAT-Mediated Inflammatory Signaling Cascade Drives Oncogenesis In AF10-Rearranged AML]]></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/10.1101/2020.08.31.275834v1?rss=1">
<title>
<![CDATA[
Attenuation of homeostatic signaling from apoptotic thymocytes triggers a global regenerative response in the thymus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.31.275834v1?rss=1</link>
<description><![CDATA[
The molecular triggers of organotypic tissue repair are unknown. The thymus, which is the primary site of T cell development, is a model of tissue damage and regeneration as it is particularly sensitive to insult, but also has a remarkable capacity for repair. However, acute and profound damage, such as that caused by common cytoreductive therapies or age-related decline, lead to involution of the thymus and prolonged T cell deficiency, precipitating life-threatening infections and malignant relapse. Consequently, there is an unmet need to boost thymic function and enhance T cell immunity. Here, we demonstrate an innate trigger of the reparative response in the thymus, centered on the attenuation of signaling directly downstream of apoptotic cell detection as thymocytes are depleted after acute damage. We found that the intracellular pattern recognition receptor NOD2, via induction of microRNA-29c, suppressed the induction of the regenerative factors IL-23 and BMP4, from thymic dendritic cells (DCs) and endothelial cells (ECs), respectively. During steady-state, when a high proportion of thymocytes are undergoing apoptosis (as a consequence of selection events during T cell development), this suppressive pathway is constitutively activated by the detection of exposed phosphatidylserine on apoptotic thymocytes by cell surface TAM receptors on DCs and ECs, with subsequent downstream activation of the Rho GTPase Rac1. However, after damage, when profound cell depletion occurs across the thymus, the TAM-Rac1-NOD2-miR29c pathway is abrogated, therefore triggering the increase in IL-23 and BMP4 levels. Importantly, this pathway could be modulated pharmacologically by inhibiting Rac1 GTPase activation with the small molecule inhibitor EHT1864, leading to increased thymic function and T cell recovery after acute damage. In conclusion, our work not only represents a novel regenerative strategy for restoring immune competence in patients whose thymic function has been compromised due to cytoreductive conditioning, infection, or age; but also, identifies a mechanism by which tissue regenerative responses are triggered.
]]></description>
<dc:creator>Kinsella, S.</dc:creator>
<dc:creator>Evandy, C. A.</dc:creator>
<dc:creator>Cooper, K.</dc:creator>
<dc:creator>Iovino, L.</dc:creator>
<dc:creator>deRoos, P. C.</dc:creator>
<dc:creator>Hopwo, K. S.</dc:creator>
<dc:creator>Granadier, D. W.</dc:creator>
<dc:creator>Smith, C. W.</dc:creator>
<dc:creator>Rafii, S.</dc:creator>
<dc:creator>Dudakov, J. A.</dc:creator>
<dc:date>2020-09-01</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.275834</dc:identifier>
<dc:title><![CDATA[Attenuation of homeostatic signaling from apoptotic thymocytes triggers a global regenerative response in the thymus]]></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/10.1101/2020.09.04.282418v1?rss=1">
<title>
<![CDATA[
Single-cell analysis of chromatin silencing programs in developmental and tumor progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.04.282418v1?rss=1</link>
<description><![CDATA[
Single-cell analysis has become a powerful approach for the molecular characterization of complex tissues. Methods for quantifying gene expression1 and chromatin accessibility2 of single cells are now well-established, but analysis of chromatin regions with specific histone modifications has been technically challenging. Here, we adapt the recently published CUT&Tag method3 to scalable single-cell platforms to profile chromatin landscapes in single cells (scCUT&Tag) from complex tissues. We focus on profiling Polycomb Group (PcG) silenced regions marked by H3K27 trimethylation (H3K27me3) in single cells as an orthogonal approach to chromatin accessibility for identifying cell states. We show that scCUT&Tag profiling of H3K27me3 distinguishes cell types in human blood and allows the generation of cell-type-specific PcG landscapes from heterogeneous tissues. Furthermore, we use scCUT&Tag to profile H3K27me3 in a brain tumor patient before and after treatment, identifying cell types in the tumor microenvironment and heterogeneity in PcG activity in the primary sample and after treatment.
]]></description>
<dc:creator>Wu, S. J.</dc:creator>
<dc:creator>Furlan, S. N.</dc:creator>
<dc:creator>Mihalas, A. B.</dc:creator>
<dc:creator>Kaya-Okur, H.</dc:creator>
<dc:creator>Feroze, A. H.</dc:creator>
<dc:creator>Emerson, S. N.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Carson, K.</dc:creator>
<dc:creator>Cimino, P. J.</dc:creator>
<dc:creator>Keene, C. D.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:creator>Sarthy, J. F.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Patel, A. P.</dc:creator>
<dc:date>2020-09-04</dc:date>
<dc:identifier>doi:10.1101/2020.09.04.282418</dc:identifier>
<dc:title><![CDATA[Single-cell analysis of chromatin silencing programs in developmental and tumor progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.04.283812v1?rss=1">
<title>
<![CDATA[
BayesSpace enables the robust characterization of spatial gene expression architecture in tissue sections at increased resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.04.283812v1?rss=1</link>
<description><![CDATA[
Recently developed spatial gene expression technologies such as the Spatial Transcriptomics and Visium platforms allow for comprehensive measurement of transcriptomic profiles while retaining spatial context. However, existing methods for analyzing spatial gene expression data often do not efficiently leverage the spatial information and fail to address the limited resolution of the technology. Here, we introduce BayesSpace, a fully Bayesian statistical method for clustering analysis and resolution enhancement of spatial transcriptomics data that seamlessly integrates into current transcriptomics analysis workflows. We show that BayesSpace improves the identification of transcriptionally distinct tissues from spatial transcriptomics samples of the brain, of melanoma, and of squamous cell carcinoma. In particular, BayesSpaces improved resolution allows the identification of tissue structure that is not detectable at the original resolution and thus not recovered by other methods. Using an in silico dataset constructed from scRNA-seq, we demonstrate that BayesSpace can spatially resolve expression patterns to near single-cell resolution without the need for external single-cell sequencing data. In all, our results illustrate the utility BayesSpace has in facilitating the discovery of biological insights from a variety of spatial transcriptomics datasets.
]]></description>
<dc:creator>Zhao, E.</dc:creator>
<dc:creator>Stone, M. R.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Pulliam, T.</dc:creator>
<dc:creator>Nghiem, P.</dc:creator>
<dc:creator>Bielas, J. H.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:date>2020-09-05</dc:date>
<dc:identifier>doi:10.1101/2020.09.04.283812</dc:identifier>
<dc:title><![CDATA[BayesSpace enables the robust characterization of spatial gene expression architecture in tissue sections at increased resolution]]></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/10.1101/2020.09.07.286880v1?rss=1">
<title>
<![CDATA[
Joint Microbial and Metabolomic Network Estimation with the Censored Gaussian Graphical Model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.07.286880v1?rss=1</link>
<description><![CDATA[
Joint analysis of microbiome and metabolomic data represents an imperative objective as the field moves beyond basic microbiome association studies and turns towards mechanistic and translational investigations. We present a censored Gaussian graphical model framework, where the metabolomic data are treated as continuous and the microbiome data as censored at zero, to identify direct interactions (defined as conditional dependence relationships) between microbial species and metabolites. Simulated examples show that our method metaMint performs favorably compared to existing ones. metaMint also provides interpretable microbe-metabolite interactions when applied to a bacterial vaginosis data set. R implementation of metaMint is available on GitHub.
]]></description>
<dc:creator>Ma, J.</dc:creator>
<dc:date>2020-09-09</dc:date>
<dc:identifier>doi:10.1101/2020.09.07.286880</dc:identifier>
<dc:title><![CDATA[Joint Microbial and Metabolomic Network Estimation with the Censored Gaussian Graphical Model]]></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/10.1101/2020.09.08.287482v1?rss=1">
<title>
<![CDATA[
Antibody binding to SARS-CoV-2 S glycoprotein correlates with, but does not predict neutralization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.08.287482v1?rss=1</link>
<description><![CDATA[
Convalescent plasma from SARS-CoV-2 infected individuals and monoclonal antibodies were shown to potently neutralize viral and pseudoviral particles carrying the S glycoprotein. However, a non-negligent proportion of plasma samples from infected individuals as well as S-specific monoclonal antibodies were reported to be non-neutralizing despite efficient interaction with the S glycoprotein in different biochemical assays using soluble recombinant forms of S or when expressed at the cell surface. How neutralization relates to binding of S glycoprotein in the context of viral particles remains to be established. Here we developed a pseudovirus capture assay (VCA) to measure the capacity of plasma samples or antibodies immobilized on ELISA plates to bind to membrane-bound S glycoproteins from SARS-CoV-2 expressed at the surface of lentiviral particles. By performing VCA and neutralization assays we observed a strong correlation between these two parameters. However, while we found that plasma samples unable to capture viral particles did not neutralize, capture did not guarantee neutralization, indicating that the capacity of antibodies to bind to the S glycoprotein at the surface of viral particles is required but not sufficient to mediate neutralization. Altogether, our results highlights the importance of better understanding the inactivation of S by plasma and neutralizing antibodies.
]]></description>
<dc:creator>Ding, S.</dc:creator>
<dc:creator>Laumaea, A.</dc:creator>
<dc:creator>Gasser, R.</dc:creator>
<dc:creator>Medjahed, H.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>McGuire, A.</dc:creator>
<dc:creator>Bazin, R.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:date>2020-09-08</dc:date>
<dc:identifier>doi:10.1101/2020.09.08.287482</dc:identifier>
<dc:title><![CDATA[Antibody binding to SARS-CoV-2 S glycoprotein correlates with, but does not predict neutralization]]></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/10.1101/2020.09.10.292078v1?rss=1">
<title>
<![CDATA[
Complete mapping of mutations to the SARS-CoV-2 spike receptor-binding domain that escape antibody recognition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.10.292078v1?rss=1</link>
<description><![CDATA[
Antibodies targeting the SARS-CoV-2 spike receptor-binding domain (RBD) are being developed as therapeutics and make a major contribution to the neutralizing antibody response elicited by infection. Here, we describe a deep mutational scanning method to map how all amino-acid mutations in the RBD affect antibody binding, and apply this method to 10 human monoclonal antibodies. The escape mutations cluster on several surfaces of the RBD that broadly correspond to structurally defined antibody epitopes. However, even antibodies targeting the same RBD surface often have distinct escape mutations. The complete escape maps predict which mutations are selected during viral growth in the presence of single antibodies, and enable us to design escape-resistant antibody cocktails-including cocktails of antibodies that compete for binding to the same surface of the RBD but have different escape mutations. Therefore, complete escape-mutation maps enable rational design of antibody therapeutics and assessment of the antigenic consequences of viral evolution.
]]></description>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Gilchuk, P.</dc:creator>
<dc:creator>Zost, S. J.</dc:creator>
<dc:creator>Binshtein, E.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Hilton, S. K.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Eguia, R.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Nargi, R. S.</dc:creator>
<dc:creator>Sutton, R. E.</dc:creator>
<dc:creator>Suryadevara, N.</dc:creator>
<dc:creator>Rothlauf, P. W.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Whelan, S. P.</dc:creator>
<dc:creator>Carnahan, R. H.</dc:creator>
<dc:creator>Crowe, J. E.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-09-10</dc:date>
<dc:identifier>doi:10.1101/2020.09.10.292078</dc:identifier>
<dc:title><![CDATA[Complete mapping of mutations to the SARS-CoV-2 spike receptor-binding domain that escape antibody recognition]]></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/10.1101/2020.09.21.306837v1?rss=1">
<title>
<![CDATA[
Baseline T cell immune phenotypes predict virologic and disease control upon SARS-CoV infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.21.306837v1?rss=1</link>
<description><![CDATA[
The COVID-19 pandemic has revealed that infection with SARS-CoV-2 can result in a wide range of clinical outcomes in humans, from asymptomatic or mild disease to severe disease that can require mechanical ventilation. An incomplete understanding of immune correlates of protection represents a major barrier to the design of vaccines and therapeutic approaches to prevent infection or limit disease. This deficit is largely due to the lack of prospectively collected, pre-infection samples from indiviuals that go on to become infected with SARS-CoV-2. Here, we utilized data from a screen of genetically diverse mice from the Collaborative Cross (CC) infected with SARS-CoV to determine whether circulating baseline T cell signatures are associated with a lack of viral control and severe disease upon infection. SARS-CoV infection of CC mice results in a variety of viral load trajectories and disease outcomes. Further, early control of virus in the lung correlates with an increased abundance of activated CD4 and CD8 T cells and regulatory T cells prior to infections across strains. A basal propensity of T cells to express IFNg and IL17 over TNFa also correlated with early viral control. Overall, a dysregulated, pro-inflammatory signature of circulating T cells at baseline was associated with severe disease upon infection. While future studies of human samples prior to infection with SARS-CoV-2 are required, our studies in mice with SARS-CoV serve as proof of concept that circulating T cell signatures at baseline can predict clinical and virologic outcomes upon SARS-CoV infection. Identification of basal immune predictors in humans could allow for identification of individuals at highest risk of severe clinical and virologic outcomes upon infection, who may thus most benefit from available clinical interventions to restrict infection and disease.

SummaryWe used a screen of genetically diverse mice from the Collaborative Cross infected with mouse-adapted SARS-CoV in combination with comprehensive pre-infection immunophenotyping to identify baseline circulating immune correlates of severe virologic and clinical outcomes upon SARS-CoV infection.
]]></description>
<dc:creator>Graham, J.</dc:creator>
<dc:creator>Swarts, J.</dc:creator>
<dc:creator>Leist, S. R.</dc:creator>
<dc:creator>Schafer, A.</dc:creator>
<dc:creator>Menachery, V. D.</dc:creator>
<dc:creator>Gralinski, L.</dc:creator>
<dc:creator>Jeng, S.</dc:creator>
<dc:creator>Miller, D. R.</dc:creator>
<dc:creator>Mooney, M.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Ferris, M. T.</dc:creator>
<dc:creator>Pardo-Manuel de Villena, F.</dc:creator>
<dc:creator>Heise, M. T.</dc:creator>
<dc:creator>Baric, R. S.</dc:creator>
<dc:creator>Lund, J. M.</dc:creator>
<dc:date>2020-09-21</dc:date>
<dc:identifier>doi:10.1101/2020.09.21.306837</dc:identifier>
<dc:title><![CDATA[Baseline T cell immune phenotypes predict virologic and disease control upon SARS-CoV infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.24.312066v1?rss=1">
<title>
<![CDATA[
Development of antibody-dependent cell cytotoxicity function in HIV-1 antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.24.312066v1?rss=1</link>
<description><![CDATA[
A prerequisite for the design of an HIV vaccine that elicits protective antibodies is understanding the developmental pathways that result in desirable antibody features. The development of antibodies that mediate antibody-dependent cellular cytotoxicity (ADCC) is particularly relevant because such antibodies have been associated with HIV protection in humans. We reconstructed the developmental pathways of six human HIV-specific ADCC antibodies using longitudinal antibody sequencing data. Most of the inferred naive antibodies did not mediate detectable ADCC. Gain of antigen binding and ADCC function typically required mutations in complementarity determining regions of one or both chains. Enhancement of ADCC potency often required additional mutations in framework regions. Antigen binding affinity and ADCC activity were correlated, but affinity alone was not sufficient to predict ADCC potency. Thus, elicitation of human ADCC antibodies may require mutations that first enable high affinity antigen recognition, followed by mutations that optimize factors contributing to functional ADCC activity.
]]></description>
<dc:creator>Doepker, L. E.</dc:creator>
<dc:creator>Danon, S.</dc:creator>
<dc:creator>Harkins, E.</dc:creator>
<dc:creator>Ralph, D.</dc:creator>
<dc:creator>Yaffe, Z.</dc:creator>
<dc:creator>Dhar, A.</dc:creator>
<dc:creator>Wagner, C.</dc:creator>
<dc:creator>Stumpf, M.</dc:creator>
<dc:creator>Arenz, D.</dc:creator>
<dc:creator>Williams, J. A.</dc:creator>
<dc:creator>Jaoko, W.</dc:creator>
<dc:creator>Mandaliya, K.</dc:creator>
<dc:creator>Lee, K. K.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2020-09-25</dc:date>
<dc:identifier>doi:10.1101/2020.09.24.312066</dc:identifier>
<dc:title><![CDATA[Development of antibody-dependent cell cytotoxicity function in HIV-1 antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.26.312348v1?rss=1">
<title>
<![CDATA[
An optimal set of inhibitors for Reverse Engineering via Kinase Regularization. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.26.312348v1?rss=1</link>
<description><![CDATA[
We present a comprehensive resource of 257 kinase inhibitor profiles against 365 human protein kinases using gold-standard kinase activity assays. We show the utility of this dataset with an improved version of Kinome Regularization (KiR) to deconvolve protein kinases involved in a cellular phenotype. We assayed protein kinase inhibitors against more than 70% of the human protein kinome and chose an optimal subset of 58 inhibitors to assay at ten doses across four orders of magnitude. We demonstrate the effectiveness of KiR to identify key kinases by using a quantitative cell migration assay and updated machine learning methods. This approach can be widely applied to biological problems for which a quantitative phenotype can be measured and which can be perturbed with our set of kinase inhibitors.
]]></description>
<dc:creator>Rata, S.</dc:creator>
<dc:creator>Gruver, J. S.</dc:creator>
<dc:creator>Trikoz, N.</dc:creator>
<dc:creator>Lukyanov, A.</dc:creator>
<dc:creator>Vultaggio, J.</dc:creator>
<dc:creator>Ceribelli, M.</dc:creator>
<dc:creator>Thomas, C.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:creator>Kirschner, M. W.</dc:creator>
<dc:creator>Peshkin, L.</dc:creator>
<dc:date>2020-09-28</dc:date>
<dc:identifier>doi:10.1101/2020.09.26.312348</dc:identifier>
<dc:title><![CDATA[An optimal set of inhibitors for Reverse Engineering via Kinase Regularization.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.28.317503v1?rss=1">
<title>
<![CDATA[
A rapidly evolving actin mediates fertility and developmental tradeoffs in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.28.317503v1?rss=1</link>
<description><![CDATA[
Most actin-related proteins (Arps) are highly conserved in eukaryotes, where they carry out well-defined cellular functions. Drosophila and mammals also encode divergent non-canonical Arps in their male-germline whose roles remain unknown. Here, we show that Arp53D, a rapidly-evolving Drosophila Arp, localizes to fusomes and actin cones, two male germline-specific actin structures critical for sperm maturation, via its non-canonical N-terminal tail. Although we expected that Arp53D loss would reduce male fertility, we instead find that Arp53D-KO males are more fertile, both in isolation and in competition with wildtype males. Upon investigating why evolution would retain a gene that negatively impacts male fertility, we unexpectedly found that Arp53D-KO females are less fertile. Moreover, KO embryos exhibit reduced viability, which worsens under heat stress. We conclude that  testis-specific Arp53D is detrimental to male fertility, but is required for female fertility and early development, leading to its long-term retention and recurrent adaptation in Drosophila.
]]></description>
<dc:creator>Schroeder, C. M.</dc:creator>
<dc:creator>Tomlin, S. A.</dc:creator>
<dc:creator>Valenzuela, J. R.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2020-09-28</dc:date>
<dc:identifier>doi:10.1101/2020.09.28.317503</dc:identifier>
<dc:title><![CDATA[A rapidly evolving actin mediates fertility and developmental tradeoffs in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.28.317578v1?rss=1">
<title>
<![CDATA[
AcrIIA22 is a novel anti-CRISPR that impairs SpyCas9 activity by relieving DNA torsion of target plasmids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.28.317578v1?rss=1</link>
<description><![CDATA[
To overcome CRISPR-Cas defense systems, many phages and mobile genetic elements encode CRISPR-Cas inhibitors called anti-CRISPRs (Acrs). Nearly all characterized Acrs directly bind Cas proteins to inactivate CRISPR immunity. Here, using functional metagenomic selection, we describe AcrIIA22, an unconventional Acr found in hypervariable genomic regions of clostridial bacteria and their prophages from human gut microbiomes. AcrIIA22 does not bind strongly to SpyCas9 but nonetheless potently inhibits its activity against plasmids. To gain insight into its mechanism, we obtained an X-ray crystal structure of AcrIIA22, which revealed homology to PC4-like nucleic-acid binding proteins. Based on mutational analyses and functional assays, we deduced that acrIIA22 encodes a DNA nickase that relieves torsional stress in supercoiled plasmids. This may render them less susceptible to SpyCas9, which uses free energy from negative supercoils to form stable R-loops. Modifying DNA topology may provide an additional route to CRISPR-Cas resistance in phages and mobile genetic elements.
]]></description>
<dc:creator>Forsberg, K. J.</dc:creator>
<dc:creator>Schmidtke, D. T.</dc:creator>
<dc:creator>Werther, R.</dc:creator>
<dc:creator>Hausman, D.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>Kaiser, B. K.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2020-09-29</dc:date>
<dc:identifier>doi:10.1101/2020.09.28.317578</dc:identifier>
<dc:title><![CDATA[AcrIIA22 is a novel anti-CRISPR that impairs SpyCas9 activity by relieving DNA torsion of target plasmids]]></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/10.1101/2020.09.30.321158v1?rss=1">
<title>
<![CDATA[
Modulation of MHC-E transport by viral decoy ligands is required for RhCMV/SIV vaccine efficacy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.30.321158v1?rss=1</link>
<description><![CDATA[
Strain 68-1 rhesus cytomegalovirus (RhCMV) vectors expressing simian immunodeficiency virus (SIV) antigens elicit CD8+ T cells that recognize peptide epitopes presented by major histocompatibility complex (MHC)-II and MHC-E molecules, instead of MHC-Ia, and are uniquely able to mediate stringent control and subsequent clearance of highly pathogenic SIV in [~]50% of vaccinated rhesus macaques (RMs). We show that the MHC-E ligand VMAPRTLLL (VL9), encoded by the Rh67 gene (or its HCMV UL40 counterpart) is required for recognition of RhCMV-infected fibroblasts by MHC-E-restricted CD8+ T cells via its ability to promote intracellular MHC-E transport. Moreover, deletion of Rh67 from 68-1 RhCMV/SIV vectors, or mutation of its embedded VL9 ligand, abrogated induction of MHC-E-restricted CD8+ T cell responses, leaving responses that exclusively target MHC-II-restricted epitopes. These MHC-II-presented CD8+ T cell responses, though comparable in response magnitude and functional differentiation to responses arising from the efficacious 68-1 vector, did not protect RMs against SIV challenge, indicating that Rh67/UL40-enabled direct priming of MHC-E-targeted CD8+ T cells is a crucial element of RhCMV/SIV vaccine efficacy.

One Sentence SummaryA cytomegalovirus protein (Rh67/UL40) that upregulates MHC-E expression on RhCMV/SIV-vector infected cells is required for induction of MHC-E-restricted CD8+ T cells and for protection against SIV.
]]></description>
<dc:creator>Verweij, M.</dc:creator>
<dc:creator>Hansen, S. G.</dc:creator>
<dc:creator>Iyer, R.</dc:creator>
<dc:creator>John, N.</dc:creator>
<dc:creator>Malouli, D.</dc:creator>
<dc:creator>Morrow, D.</dc:creator>
<dc:creator>Scholz, I.</dc:creator>
<dc:creator>Womack, J.</dc:creator>
<dc:creator>Abdulhaqq, S.</dc:creator>
<dc:creator>Gilbride, R. M.</dc:creator>
<dc:creator>Hughes, C. M.</dc:creator>
<dc:creator>Ventura, A. B.</dc:creator>
<dc:creator>Ford, J. C.</dc:creator>
<dc:creator>Selseth, A. N.</dc:creator>
<dc:creator>Oswald, K.</dc:creator>
<dc:creator>Shoemaker, R.</dc:creator>
<dc:creator>Berkemeier, B.</dc:creator>
<dc:creator>Borsche, W. J.</dc:creator>
<dc:creator>Hull, M.</dc:creator>
<dc:creator>Shao, J.</dc:creator>
<dc:creator>Sacha, J.</dc:creator>
<dc:creator>Axthelm, M. K.</dc:creator>
<dc:creator>Edlefsen, P.</dc:creator>
<dc:creator>Lifson, J. D.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:creator>Früh, K.</dc:creator>
<dc:date>2020-10-02</dc:date>
<dc:identifier>doi:10.1101/2020.09.30.321158</dc:identifier>
<dc:title><![CDATA[Modulation of MHC-E transport by viral decoy ligands is required for RhCMV/SIV vaccine efficacy]]></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/10.1101/2020.09.30.321349v1?rss=1">
<title>
<![CDATA[
Cytomegaloviral determinants of CD8+ T cell programming and RhCMV/SIV vaccine efficacy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.30.321349v1?rss=1</link>
<description><![CDATA[
Simian immunodeficiency virus (SIV) insert-expressing, 68-1 Rhesus Cytomegalovirus (RhCMV/SIV) vectors elicit major histocompatibility complex (MHC)-E- and -II-restricted, SIV-specific CD8+ T cell responses, but the basis of these unconventional responses and their contribution to demonstrated vaccine efficacy against SIV challenge in the rhesus monkeys (RMs) has not been characterized. We demonstrate that these unconventional responses resulted from a chance genetic rearrangement in 68-1 RhCMV that abrogated the function of eight distinct immunomodulatory gene products encoded in two RhCMV genomic regions (Rh157.5/.4 and Rh158-161). Differential repair of these genes with either RhCMV-derived or orthologous human CMV (HCMV)-derived sequences (UL128/130; UL146/147) leads to either of two distinct CD8+ T cell response types - MHC-Ia-restricted-only, or a mix of MHC-II- and MHC-Ia-restricted CD8+ T cells. Despite response magnitude and functional differentiation being similar to RhCMV 68-1, neither alternative response type mediated protection against SIV challenge. These findings implicate MHC-E-restricted CD8+ T cell responses as mediators of anti-SIV efficacy and indicate that translation of RhCMV/SIV vector efficacy to humans will likely require deletion of all the genes that inhibit these responses from the HCMV/HIV vector.

One-sentence summaryEight genes in two spatially distinct RhCMV gene regions control induction of unconventionally restricted CD8+ T cell responses and the efficacy of RhCMV/SIV vaccine vectors against SIV challenge.
]]></description>
<dc:creator>Malouli, D.</dc:creator>
<dc:creator>Hansen, S. G.</dc:creator>
<dc:creator>Hancock, M. H.</dc:creator>
<dc:creator>Hughes, C. M.</dc:creator>
<dc:creator>Ford, J. C.</dc:creator>
<dc:creator>Gilbride, R. M.</dc:creator>
<dc:creator>Ventura, A. B.</dc:creator>
<dc:creator>Morrow, D.</dc:creator>
<dc:creator>Taher, H.</dc:creator>
<dc:creator>Uebelhoer, L. S.</dc:creator>
<dc:creator>McArdle, M. R.</dc:creator>
<dc:creator>Papen, C. R.</dc:creator>
<dc:creator>Espinosa Trethewy, R.</dc:creator>
<dc:creator>Oswald, K.</dc:creator>
<dc:creator>Shoemaker, R.</dc:creator>
<dc:creator>Berkemeier, B.</dc:creator>
<dc:creator>Bosche, W.</dc:creator>
<dc:creator>Hull, M.</dc:creator>
<dc:creator>Green, J.</dc:creator>
<dc:creator>Axthelm, M. K.</dc:creator>
<dc:creator>Shao, J.</dc:creator>
<dc:creator>Edlefsen, P.</dc:creator>
<dc:creator>Grey, F.</dc:creator>
<dc:creator>Nelson, J. A.</dc:creator>
<dc:creator>Lifson, J. D.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Sacha, J.</dc:creator>
<dc:creator>Früh, K.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:date>2020-10-01</dc:date>
<dc:identifier>doi:10.1101/2020.09.30.321349</dc:identifier>
<dc:title><![CDATA[Cytomegaloviral determinants of CD8+ T cell programming and RhCMV/SIV vaccine efficacy]]></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/10.1101/2020.10.06.328518v1?rss=1">
<title>
<![CDATA[
Helicobacter pylori accelerates KRAS-dependent gastric dysplasia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.06.328518v1?rss=1</link>
<description><![CDATA[
More than 80% of gastric cancer is attributable to stomach infection with Helicobacter pylori (Hp), even though the bacterium is not always present at time of diagnosis. Infection is thought to lead to cancer by promoting the accumulation of oncogenic mutations downstream of inflammation; once oncogenic pathways become activated, infection may become dispensable for cancer development. Gastric preneoplastic progression involves sequential changes to the tissue, including loss of parietal cells, spasmolytic polypeptide-expressing metaplasia (SPEM), intestinal metaplasia (IM) and dysplasia. In mice, active KRAS expression recapitulates these tissue changes in the absence of Hp infection. This model provides an experimental system to investigate whether Hp infection has additional roles in preneoplastic progression, beyond initiating inflammation. Mice were assessed by evaluating tissue histology, gene expression changes, the immune cell repertoire, and expression of metaplasia and dysplasia markers. Compared to Hp-/KRAS+ mice, Hp+/KRAS+ mice had i) severe T cell infiltration and altered macrophage polarization; ii) altered expression of metaplasia markers, including increased expression of CD44v9 (SPEM) and decreased expression of TFF3 (IM); iii) more dysplastic (TROP2+) glands; and iv) greater proliferation of metaplastic and dysplastic glands. Hp was able to persistently colonize the stomach during the onset of these tissue changes, and eradication of Hp with antibiotics prevented metaplastic, dysplastic and proliferation marker changes. Collectively, these results suggest that gastric preneoplastic progression differs between Hp+ and Hp-cases, and that sustained Hp infection can promote the later stages of gastric preneoplastic progression, in addition to its established role in initiating chronic inflammation.
]]></description>
<dc:creator>O'Brien, V. P.</dc:creator>
<dc:creator>Koehne, A.</dc:creator>
<dc:creator>Dubrulle, J.</dc:creator>
<dc:creator>Rodriguez, A.</dc:creator>
<dc:creator>Leverich, C.</dc:creator>
<dc:creator>Kong, P.</dc:creator>
<dc:creator>Campbell, J.</dc:creator>
<dc:creator>Pierce, R.</dc:creator>
<dc:creator>Goldenring, J.</dc:creator>
<dc:creator>Choi, E.</dc:creator>
<dc:creator>Salama, N.</dc:creator>
<dc:date>2020-10-06</dc:date>
<dc:identifier>doi:10.1101/2020.10.06.328518</dc:identifier>
<dc:title><![CDATA[Helicobacter pylori accelerates KRAS-dependent gastric dysplasia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.06.328948v1?rss=1">
<title>
<![CDATA[
Automated CUT&Tag profiling of chromatin heterogeneity in mixed-lineage leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.06.328948v1?rss=1</link>
<description><![CDATA[
Acute myeloid and lymphoid leukemias often harbor chromosomal translocations involving the Mixed Lineage Leukemia-1 gene, encoding the KMT2A lysine methyltransferase. The most common translocations produce in-frame fusions of KMT2A to other chromatin regulatory proteins. Here we develop a strategy to map the genome-wide occupancy of oncogenic KMT2A fusion proteins in primary patient samples regardless of fusion partner. By modifying the versatile CUT&Tag method for full automation we identify common and tumor-specific patterns of aberrant chromatin regulation induced by different KMT2A fusion proteins. Integration of automated and single-cell CUT&Tag uncovers epigenomic heterogeneity within patient samples and predicts sensitivity to therapeutic agents.
]]></description>
<dc:creator>Janssens, D.</dc:creator>
<dc:creator>Meers, M. P.</dc:creator>
<dc:creator>Wu, S. J.</dc:creator>
<dc:creator>Babaeva, E.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Sarthy, J. F.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2020-10-08</dc:date>
<dc:identifier>doi:10.1101/2020.10.06.328948</dc:identifier>
<dc:title><![CDATA[Automated CUT&Tag profiling of chromatin heterogeneity in mixed-lineage leukemia]]></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/10.1101/2020.10.08.330688v1?rss=1">
<title>
<![CDATA[
CD8+ T cell responses in convalescent COVID-19 individuals target epitopes from the entireSARS-CoV-2 proteome and show kinetics of early differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.08.330688v1?rss=1</link>
<description><![CDATA[
Characterization of the T cell response in individuals who recover from SARS-CoV-2 infection is critical to understanding its contribution to protective immunity. A multiplexed peptide-MHC tetramer approach was used to screen 408 SARS-CoV-2 candidate epitopes for CD8+ T cell recognition in a cross-sectional sample of 30 COVID-19 convalescent individuals. T cells were evaluated using a 28-marker phenotypic panel, and findings were modelled against time from diagnosis, humoral and inflammatory responses. 132 distinct SARS-CoV-2-specific CD8+ T cell epitope responses across six different HLAs were detected, corresponding to 52 unique reactivities. T cell responses were directed against several structural and non-structural virus proteins. Modelling demonstrated a coordinated and dynamic immune response characterized by a decrease in inflammation, increase in neutralizing antibody titer, and differentiation of a specific CD8+ T cell response. Overall, T cells exhibited distinct differentiation into stem-cell and transitional memory states, subsets, which may be key to developing durable protection.
]]></description>
<dc:creator>Kared, H.</dc:creator>
<dc:creator>Redd, A. D.</dc:creator>
<dc:creator>Bloch, E. M.</dc:creator>
<dc:creator>Bonny, T. S.</dc:creator>
<dc:creator>Sumatoh, H.</dc:creator>
<dc:creator>Kairi, F.</dc:creator>
<dc:creator>Carbajo, D.</dc:creator>
<dc:creator>Abel, B.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Bettinotti, M. P.</dc:creator>
<dc:creator>Benner, S. E.</dc:creator>
<dc:creator>Patel, E. U.</dc:creator>
<dc:creator>Littlefield, K.</dc:creator>
<dc:creator>Laeyendecker, O.</dc:creator>
<dc:creator>Shoham, S.</dc:creator>
<dc:creator>Sullivan, D.</dc:creator>
<dc:creator>Casadevall, A.</dc:creator>
<dc:creator>Pekosz, A.</dc:creator>
<dc:creator>Nardin, A.</dc:creator>
<dc:creator>Fehlings, M.</dc:creator>
<dc:creator>Tobian, A. A.</dc:creator>
<dc:creator>Quinn, T. C.</dc:creator>
<dc:date>2020-10-08</dc:date>
<dc:identifier>doi:10.1101/2020.10.08.330688</dc:identifier>
<dc:title><![CDATA[CD8+ T cell responses in convalescent COVID-19 individuals target epitopes from the entireSARS-CoV-2 proteome and show kinetics of early differentiation]]></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/10.1101/2020.10.09.331900v1?rss=1">
<title>
<![CDATA[
HP1α is a chromatin crosslinker that controls nuclear and mitotic chromosome mechanics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.09.331900v1?rss=1</link>
<description><![CDATA[
Chromatin, which consists of DNA and associated proteins, contains genetic information and is a mechanical component of the nucleus. Heterochromatic histone methylation controls nucleus and chromosome stiffness, but the contribution of heterochromatin protein HP1 (CBX5) is unknown. We used a novel HP1 auxin-inducible degron human cell line to rapidly degrade HP1. Degradation did not alter transcription, local chromatin compaction, or histone methylation, but did decrease chromatin stiffness. Single-nucleus micromanipulation reveals that HP1 is essential to chromatin-based mechanics and maintains nuclear morphology, separate from histone methylation. Further experiments with dimerization-deficient HP1I165E indicate that chromatin crosslinking via HP1 dimerization is critical, while polymer simulations demonstrate the importance of chromatin-chromatin crosslinkers in mechanics. In mitotic chromosomes, HP1 similarly bolsters stiffness while aiding in mitotic alignment and faithful segregation. HP1 is therefore a critical chromatin-crosslinking protein that provides mechanical strength to chromosomes and the nucleus throughout the cell cycle and supports cellular functions.
]]></description>
<dc:creator>Strom, A. R.</dc:creator>
<dc:creator>Biggs, R. J.</dc:creator>
<dc:creator>Banigan, E. J.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Chiu, K.</dc:creator>
<dc:creator>Herman, C.</dc:creator>
<dc:creator>Collado, J.</dc:creator>
<dc:creator>Yue, F.</dc:creator>
<dc:creator>Ritland Politz, J. C.</dc:creator>
<dc:creator>Tait, L. J.</dc:creator>
<dc:creator>Scalzo, D.</dc:creator>
<dc:creator>Telling, A.</dc:creator>
<dc:creator>Groudine, M.</dc:creator>
<dc:creator>Brangwynne, C. P.</dc:creator>
<dc:creator>Marko, J.</dc:creator>
<dc:creator>Stephens, A. D.</dc:creator>
<dc:date>2020-10-09</dc:date>
<dc:identifier>doi:10.1101/2020.10.09.331900</dc:identifier>
<dc:title><![CDATA[HP1α is a chromatin crosslinker that controls nuclear and mitotic chromosome mechanics]]></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/10.1101/2020.10.09.333971v1?rss=1">
<title>
<![CDATA[
Gene-level metagenomics identifies genome islands associated with immunotherapy response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.09.333971v1?rss=1</link>
<description><![CDATA[
Researchers must be able to generate experimentally testable hypotheses from sequencing-based observational microbiome experiments to discover the mechanisms underlying the influence of gut microbes on human health. We describe a novel bioinformatics tool for identifying testable hypotheses based on gene-level metagenomic analysis of WGS microbiome data (geneshot). By applying geneshot to two independent previously published cohorts, we identified microbial genomic islands consistently associated with response to immune checkpoint inhibitor (ICI)-based cancer treatment in culturable type strains. The identified genomic islands are within operons involved in type II secretion, TonB-dependent transport, and bacteriophage growth. These results, as well as the underlying methodology, inform further mechanistic studies and facilitate the development of microbiome-enhanced therapeutics.
]]></description>
<dc:creator>Minot, S. S.</dc:creator>
<dc:creator>Barry, K. C.</dc:creator>
<dc:creator>Kasman, C.</dc:creator>
<dc:creator>Golob, J. L.</dc:creator>
<dc:creator>Willis, A. D.</dc:creator>
<dc:date>2020-10-10</dc:date>
<dc:identifier>doi:10.1101/2020.10.09.333971</dc:identifier>
<dc:title><![CDATA[Gene-level metagenomics identifies genome islands associated with immunotherapy response]]></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/10.1101/2020.10.10.334540v1?rss=1">
<title>
<![CDATA[
Innate behavior sequence progression by peptide-mediated interorgan crosstalk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.10.334540v1?rss=1</link>
<description><![CDATA[
Innate behaviors consist of a succession of genetically-hardwired motor and physiological subprograms that can be coupled to drastic morphogenetic changes. How these integrative responses are orchestrated is not completely understood. Here, we provide insight into these mechanisms by studying pupariation, a multi-step innate behavior of fly larvae that is critical for survival during metamorphosis. We find that the steroid-hormone ecdysone triggers parallel pupariation neuromotor and morphogenetic subprograms, which include the induction of the relaxin-peptide hormone, Dilp8, in the epidermis. Dilp8 acts on six Lgr3-positive thoracic interneurons to couple both subprograms in time and to instruct neuromotor subprogram switching during behavior. Our work reveals that interorgan feedback gates progression between subunits of an innate behavior and points to an ancestral neuromodulatory function of relaxin signaling.
]]></description>
<dc:creator>Heredia, F.</dc:creator>
<dc:creator>Volonte, Y.</dc:creator>
<dc:creator>Pereirinha, J.</dc:creator>
<dc:creator>Fernandez-Acosta, M.</dc:creator>
<dc:creator>Casimiro, A. P.</dc:creator>
<dc:creator>Belem, C. G.</dc:creator>
<dc:creator>Viegas, F.</dc:creator>
<dc:creator>Tanaka, K.</dc:creator>
<dc:creator>Arana, M.</dc:creator>
<dc:creator>Cardoso, G. A.</dc:creator>
<dc:creator>Macedo, A.</dc:creator>
<dc:creator>Kotowicz, M.</dc:creator>
<dc:creator>Prado Spalm, F. H.</dc:creator>
<dc:creator>Dibo, M.</dc:creator>
<dc:creator>Monfardini, R. D.</dc:creator>
<dc:creator>Torres, T. T.</dc:creator>
<dc:creator>Mendes, C. S.</dc:creator>
<dc:creator>Garelli, A.</dc:creator>
<dc:creator>Gontijo, A. M.</dc:creator>
<dc:date>2020-10-10</dc:date>
<dc:identifier>doi:10.1101/2020.10.10.334540</dc:identifier>
<dc:title><![CDATA[Innate behavior sequence progression by peptide-mediated interorgan crosstalk]]></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/10.1101/2020.10.12.335331v1?rss=1">
<title>
<![CDATA[
Integrated analysis of multimodal single-cell data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.12.335331v1?rss=1</link>
<description><![CDATA[
The simultaneous measurement of multiple modalities, known as multimodal analysis, represents an exciting frontier for single-cell genomics and necessitates new computational methods that can define cellular states based on multiple data types. Here, we introduce  weighted-nearest neighbor analysis, an unsupervised framework to learn the relative utility of each data type in each cell, enabling an integrative analysis of multiple modalities. We apply our procedure to a CITE-seq dataset of hundreds of thousands of human white blood cells alongside a panel of 228 antibodies to construct a multimodal reference atlas of the circulating immune system. We demonstrate that integrative analysis substantially improves our ability to resolve cell states and validate the presence of previously unreported lymphoid subpopulations. Moreover, we demonstrate how to leverage this reference to rapidly map new datasets, and to interpret immune responses to vaccination and COVID-19. Our approach represents a broadly applicable strategy to analyze single-cell multimodal datasets, including paired measurements of RNA and chromatin state, and to look beyond the transcriptome towards a unified and multimodal definition of cellular identity.

AvailabilityInstallation instructions, documentation, tutorials, and CITE-seq datasets are available at http://www.satijalab.org/seurat
]]></description>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Hao, S.</dc:creator>
<dc:creator>Andersen-Nissen, E.</dc:creator>
<dc:creator>Mauck, W. M.</dc:creator>
<dc:creator>Zheng, S.</dc:creator>
<dc:creator>Butler, A.</dc:creator>
<dc:creator>Lee, M. J.</dc:creator>
<dc:creator>Wilk, A. J.</dc:creator>
<dc:creator>Darby, C.</dc:creator>
<dc:creator>Zagar, M.</dc:creator>
<dc:creator>Hoffman, P.</dc:creator>
<dc:creator>Stoeckius, M.</dc:creator>
<dc:creator>Papalexi, E.</dc:creator>
<dc:creator>Mimitou, E. P.</dc:creator>
<dc:creator>Jain, J.</dc:creator>
<dc:creator>Srivastava, A.</dc:creator>
<dc:creator>Stuart, T.</dc:creator>
<dc:creator>Fleming, L. B.</dc:creator>
<dc:creator>Yeung, B.</dc:creator>
<dc:creator>Rogers, A. J.</dc:creator>
<dc:creator>McElrath, J. M.</dc:creator>
<dc:creator>Blish, C. A.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Smibert, P.</dc:creator>
<dc:creator>Satija, R.</dc:creator>
<dc:date>2020-10-12</dc:date>
<dc:identifier>doi:10.1101/2020.10.12.335331</dc:identifier>
<dc:title><![CDATA[Integrated analysis of multimodal single-cell data]]></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/10.1101/2020.10.15.340521v1?rss=1">
<title>
<![CDATA[
Collective nuclear behavior shapes bilateral nuclear symmetry for subsequent left-right asymmetric morphogenesis in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.15.340521v1?rss=1</link>
<description><![CDATA[
Proper organ development often requires nuclei to move to a specific position within the cell. To determine how nuclear positioning affects left-right (LR) development in the Drosophila anterior midgut (AMG), we developed a surface-modeling method to measure and describe nuclear behavior at stages 13-14, captured in three-dimensional time-lapse movies. We describe the distinctive positioning and a novel collective nuclear behavior by which nuclei align LR-symmetrically along the anterior-posterior axis in the visceral muscles that overlie the midgut and are responsible for this organs LR-asymmetric development. Wnt4 signaling is crucial for the collective behavior and proper positioning of the nuclei, as are myosin II and LINC complex, without which the nuclei failed to align LR-symmetrically. The LR-symmetric positioning of the nuclei is important for the subsequent LR-asymmetric development of the AMG. We propose that the bilaterally symmetrical positioning of these nuclei may be mechanically coupled with subsequent LR-asymmetric morphogenesis.
]]></description>
<dc:creator>Shin, D.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Morishita, Y.</dc:creator>
<dc:creator>Eiraku, M.</dc:creator>
<dc:creator>Yamakawa, T. Y.</dc:creator>
<dc:creator>Sasamura, T. S.</dc:creator>
<dc:creator>Akiyama, M. A.</dc:creator>
<dc:creator>Inaki, M.</dc:creator>
<dc:creator>Matsuno, K. M.</dc:creator>
<dc:date>2020-10-15</dc:date>
<dc:identifier>doi:10.1101/2020.10.15.340521</dc:identifier>
<dc:title><![CDATA[Collective nuclear behavior shapes bilateral nuclear symmetry for subsequent left-right asymmetric morphogenesis in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.16.334714v1?rss=1">
<title>
<![CDATA[
Loss of MGA mediated Polycomb repression promotes tumor progression and invasiveness 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.16.334714v1?rss=1</link>
<description><![CDATA[
MGA, a transcription factor and member of the MYC network, is mutated or deleted in a broad spectrum of malignancies. As a critical test of a tumor suppressive role, we inactivated Mga in two mouse models of non-small cell lung cancer using a CRISPR based approach. MGA loss significantly accelerated tumor growth in both models and led to de-repression of atypical Polycomb PRC1.6, E2F and MYC-MAX targets. Similarly, MGA depletion in human lung adenocarcinoma lines augmented invasive capabilities. We further show that MGA, E2F6 and L3MBTL2 co-occupy thousands of promoters and that MGA stabilizes PRC1.6 subunits. Lastly, we report that MGA loss has also a pro-growth effect in human colon organoids. Our studies establish MGA as a bona fide tumor suppressor in vivo and suggest a tumor suppressive mechanism in adenocarcinomas resulting from widespread transcriptional attenuation of MYC and E2F targets mediated by an atypical Polycomb complex containing MGA-MAX dimers.
]]></description>
<dc:creator>Mathsyaraja, H.</dc:creator>
<dc:creator>Catchpole, J.</dc:creator>
<dc:creator>Eastwood, E.</dc:creator>
<dc:creator>Babaeva, E.</dc:creator>
<dc:creator>Geuenich, M. J.</dc:creator>
<dc:creator>Cheng, P.-F.</dc:creator>
<dc:creator>Freie, B.</dc:creator>
<dc:creator>Ayers, J. L.</dc:creator>
<dc:creator>Yu, M.</dc:creator>
<dc:creator>Wu, N.</dc:creator>
<dc:creator>Poudel, K. R.</dc:creator>
<dc:creator>Koehne, A.</dc:creator>
<dc:creator>Grady, W.</dc:creator>
<dc:creator>Houghton, A. M.</dc:creator>
<dc:creator>Shiio, Y.</dc:creator>
<dc:creator>MacPherson, D. P.</dc:creator>
<dc:creator>Eisenman, R. N.</dc:creator>
<dc:date>2020-10-17</dc:date>
<dc:identifier>doi:10.1101/2020.10.16.334714</dc:identifier>
<dc:title><![CDATA[Loss of MGA mediated Polycomb repression promotes tumor progression and invasiveness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.18.344663v1?rss=1">
<title>
<![CDATA[
Multi-omic Single-cell Atlas Reveals Broad Ex Vivo Effects on Human Immunobiology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.18.344663v1?rss=1</link>
<description><![CDATA[
Multi-omic profiling of human peripheral blood is increasingly utilized to identify biomarkers and pathophysiologic mechanisms of disease. The importance of these platforms in clinical and translational studies led us to investigate the impact of delayed blood processing on the numbers and state of peripheral blood mononuclear cells (PBMC) and on the plasma proteome. Similar to previous studies, we show minimal effects of delayed processing on the numbers and general phenotype of PBMCs up to 18 hours. In contrast, profound changes in the single-cell transcriptome and composition of the plasma proteome become evident as early as 6 hours after blood draw. These reflect patterns of cellular activation across diverse cell types that lead to progressive distancing of the gene expression state and plasma proteome from native in vivo biology. Differences accumulating during an overnight rest (18 hours) could confound relevant biologic variance related to many underlying disease states.
]]></description>
<dc:creator>Savage, A. K.</dc:creator>
<dc:creator>Gutschow, M. V.</dc:creator>
<dc:creator>Chiang, T.</dc:creator>
<dc:creator>Henderson, K.</dc:creator>
<dc:creator>Green, R.</dc:creator>
<dc:creator>Chaudhari, M.</dc:creator>
<dc:creator>Swanson, E.</dc:creator>
<dc:creator>Heubeck, A. T.</dc:creator>
<dc:creator>Kondza, N.</dc:creator>
<dc:creator>Burley, K. C.</dc:creator>
<dc:creator>Genge, P. C.</dc:creator>
<dc:creator>Lord, C.</dc:creator>
<dc:creator>Smith, T.</dc:creator>
<dc:creator>Beaubien, A.</dc:creator>
<dc:creator>Goldy, J.</dc:creator>
<dc:creator>Bolouri, H.</dc:creator>
<dc:creator>Buckner, J. H.</dc:creator>
<dc:creator>Meijer, P.</dc:creator>
<dc:creator>Coffey, E. M.</dc:creator>
<dc:creator>Li, X.-j.</dc:creator>
<dc:creator>Skene, P. J.</dc:creator>
<dc:creator>Torgerson, T. R.</dc:creator>
<dc:creator>Bumol, T. F.</dc:creator>
<dc:date>2020-10-19</dc:date>
<dc:identifier>doi:10.1101/2020.10.18.344663</dc:identifier>
<dc:title><![CDATA[Multi-omic Single-cell Atlas Reveals Broad Ex Vivo Effects on Human Immunobiology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.21.348623v1?rss=1">
<title>
<![CDATA[
High-resolution mapping of the neutralizing and binding specificities of polyclonal rabbit serum elicited by HIV Env trimer immunization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.21.348623v1?rss=1</link>
<description><![CDATA[
Mapping the epitope specificities of polyclonal serum is critical to rational vaccine design. However, most high-resolution mapping approaches involve isolating and characterizing individual monoclonal antibodies, which incompletely defines the full polyclonal response. Here we use two complementary approaches to directly map the specificities of the neutralizing and binding antibodies of polyclonal anti-HIV-1 sera from rabbits immunized with BG505 Env SOSIP trimers. To map the neutralizing specificity, we used mutational antigenic profiling to determine how all amino-acid mutations in Env affected viral neutralization. To map the binding specificity, we used electron microscopy polyclonal epitope mapping (EMPEM) to directly visualize the Fabs in serum bound to Env trimers. Mutational antigenic profiling showed that the dominant neutralizing specificities were the C3/V5 and/or 241/289 glycan hole epitopes, which were generally only a subset of the more diverse binding specificities mapped with EMPEM. Additional differences between binding and neutralization reflected antigenicity differences between virus and soluble Env trimer. Further, mutational antigenic profiling was able to refine epitope specificity in residue-level detail directly from sera, revealing subtle differences across rabbits. Together, mutational antigenic profiling and EMPEM allow for a holistic view of the binding and neutralizing specificity of polyclonal sera and could be used to finely evaluate and guide vaccine design.
]]></description>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Pratap, P.</dc:creator>
<dc:creator>Malone, K.</dc:creator>
<dc:creator>Hilton, S. K.</dc:creator>
<dc:creator>Ketas, T.</dc:creator>
<dc:creator>Cottrell, C. A.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Moore, J. P.</dc:creator>
<dc:creator>Klasse, P. J.</dc:creator>
<dc:creator>Ward, A. B.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-10-21</dc:date>
<dc:identifier>doi:10.1101/2020.10.21.348623</dc:identifier>
<dc:title><![CDATA[High-resolution mapping of the neutralizing and binding specificities of polyclonal rabbit serum elicited by HIV Env trimer immunization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.22.351627v1?rss=1">
<title>
<![CDATA[
B LYMPHOCYTES, BUT NOT DENDRITIC CELLS, EFFICIENTLY HIV-1 TRANS-INFECT NAÏVE CD4+ T CELLS: IMPLICATIONS FOR THE VIRAL RESERVOIR 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.22.351627v1?rss=1</link>
<description><![CDATA[
Insight into the establishment and maintenance of HIV-1 infection in resting CD4+ T cell subsets is critical for the development of therapeutics targeting the HIV-1 reservoir. Although the frequency of HIV-1 infection, as quantified by the frequency of HIV-1 DNA, is lower in CD4+ naive T cells (TN) compared to the memory T cell subsets, recent studies have shown that TN cells harbor a large pool of replication-competent virus. Interestingly, however, TN cells are highly resistant to direct (cis) HIV-1 infection in vitro, in particular to R5-tropic HIV-1, as TN cells do not express CCR5. In this study, we investigated whether TN cells could be efficiently HIV-1 trans-infected by professional antigen-presenting B lymphocytes and myeloid dendritic cells (DC) in the absence of global T cell activation. We found that B cells, but not DC, have a unique ability to efficiently trans infect TN cells in vitro. In contrast, both B cells and DC mediated HIV-1 trans infection of memory and activated CD4+ T cells. Moreover, we found that TN isolated from HIV-1-infected nonprogressors (NP) harbor significantly disproportionately lower levels of HIV-1 DNA compared to TN isolated from progressors. This is consistent with our previous finding that APC derived from NP do not efficiently trans-infect CD4+ T cells due to alterations in APC cholesterol metabolism and cell membrane lipid raft organization. These findings support that B cell-mediated trans infection of TN cells with HIV-1 has a more profound role than previously considered in establishing the viral reservoir and control of HIV-1 disease progression.

ImportanceThe latent human immunodeficiency virus type 1 (HIV-1) reservoir in persons on antiretroviral therapy represents a major barrier to a cure. Although most studies have focused on the HIV-1 reservoir in the memory T cell subset, replication competent HIV-1 has been isolated from naive T cells, and CCR5-tropic HIV-1 has been recovered from CCR5negTN cells from ART-suppressed HIV-1-infected individuals. In this study, we showed that CCR5negTN cells are efficiently trans infected with R-5 tropic HIV-1 by B lymphocytes, but not by myeloid dendritic cells. Furthermore, we found that TN isolated from NP harbor no or significantly less copies of HIV-1 DNA compared to ART-suppressed progressors. These findings support that B cell-mediated trans infection of TN cells with HIV-1 has a more profound role than previously considered in establishing the viral reservoir and control of HIV-1 disease progression. Understanding the establishment and maintenance of the HIV-1 latent reservoir is fundamental for the design of effective treatments for viral eradication.
]]></description>
<dc:creator>Gerberick, A.</dc:creator>
<dc:creator>DeLucia, D. C.</dc:creator>
<dc:creator>Piazza, P. A.</dc:creator>
<dc:creator>Alaoui-El-Azher, M.</dc:creator>
<dc:creator>Rinaldo, C. R.</dc:creator>
<dc:creator>Sluis-Cremer, N.</dc:creator>
<dc:creator>Rappocciolo, G.</dc:creator>
<dc:date>2020-10-23</dc:date>
<dc:identifier>doi:10.1101/2020.10.22.351627</dc:identifier>
<dc:title><![CDATA[B LYMPHOCYTES, BUT NOT DENDRITIC CELLS, EFFICIENTLY HIV-1 TRANS-INFECT NAÏVE CD4+ T CELLS: IMPLICATIONS FOR THE VIRAL RESERVOIR]]></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/10.1101/2020.10.29.360800v1?rss=1">
<title>
<![CDATA[
Epitope profiling reveals binding signatures of SARS-CoV-2 immune response and cross-reactivity with endemic HCoVs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.29.360800v1?rss=1</link>
<description><![CDATA[
A major goal of current SARS-CoV-2 vaccine efforts is to elicit antibody responses that confer protection. Mapping the epitope targets of the SARS-CoV-2 antibody response is critical for innovative vaccine design, diagnostics, and development of therapeutics. Here, we developed a phage display library to map antibody binding sites at high resolution within the complete viral proteomes of all human-infecting coronaviruses in patients with mild or moderate/severe COVID-19. The dominant immune responses to SARS-CoV-2 were targeted to regions spanning the Spike protein, Nucleocapsid, and ORF1ab. Some epitopes were identified in the majority of samples while others were rare, and we found variation in the number of epitopes targeted by different individuals. We also identified a set of cross-reactive sequences that were bound by antibodies in SARS-CoV-2 unexposed individuals. Finally, we uncovered a subset of enriched epitopes from commonly circulating human coronaviruses with significant homology to highly reactive SARS-CoV-2 sequences.
]]></description>
<dc:creator>Stoddard, C. I.</dc:creator>
<dc:creator>Galloway, J.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Shipley, M. M.</dc:creator>
<dc:creator>Itell, H. L.</dc:creator>
<dc:creator>Wolf, C. R.</dc:creator>
<dc:creator>Logue, J. K.</dc:creator>
<dc:creator>Magedson, A.</dc:creator>
<dc:creator>Sung, K.</dc:creator>
<dc:creator>Garrett, M.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Laserson, U.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2020-10-29</dc:date>
<dc:identifier>doi:10.1101/2020.10.29.360800</dc:identifier>
<dc:title><![CDATA[Epitope profiling reveals binding signatures of SARS-CoV-2 immune response and cross-reactivity with endemic HCoVs]]></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/10.1101/2020.11.04.368597v1?rss=1">
<title>
<![CDATA[
Pruriception and neuronal coding in nociceptor subtypes in human and nonhuman primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.04.368597v1?rss=1</link>
<description><![CDATA[
In human, intradermal administration of {beta}-alanine (ALA) and bovine adrenal medulla peptide 8-22 (BAM8-22) evokes the sensation of itch. Currently, it is unknown which human dorsal root ganglion (DRG) neurons express the receptors of these pruritogens, MRGPRD and MRGPRX1 respectively, and which cutaneous afferents these pruritogens activate in primate. In situ hybridization studies revealed that MRGPRD and MRGPRX1 are co-expressed in a subpopulation of TRPV1+ human DRG neurons. In electrophysiological recordings in nonhuman primates (Macaca nemestrina), subtypes of polymodal C-fiber nociceptors are preferentially activated by ALA and BAM8-22, with significant overlap. When pruritogens ALA, BAM8-22 and histamine, that activate different subclasses of C-fiber afferents, are administered in combination, human volunteers report itch and nociceptive sensations similar to those induced by a single pruritogen. Our results provide evidence for differences in pruriceptive processing between primates and rodents, and do not support the spatial contrast theory of coding of itch and pain.
]]></description>
<dc:creator>Klein, A.</dc:creator>
<dc:creator>Solinski, H. J.</dc:creator>
<dc:creator>Malewicz, N.</dc:creator>
<dc:creator>Ieong, H. F.-h.</dc:creator>
<dc:creator>Shimada, S.</dc:creator>
<dc:creator>Hartke, T. V.</dc:creator>
<dc:creator>Wooten, M.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Hoon, M. A.</dc:creator>
<dc:creator>LaMotte, R. H.</dc:creator>
<dc:creator>Ringkamp, M.</dc:creator>
<dc:date>2020-11-04</dc:date>
<dc:identifier>doi:10.1101/2020.11.04.368597</dc:identifier>
<dc:title><![CDATA[Pruriception and neuronal coding in nociceptor subtypes in human and nonhuman primates]]></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/10.1101/2020.11.06.371047v1?rss=1">
<title>
<![CDATA[
Rapid adaptation to human protein kinase R by a unique genomic rearrangement in rhesus cytomegalovirus. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.06.371047v1?rss=1</link>
<description><![CDATA[
Cytomegaloviruses (CMVs) are generally unable to cross species barriers, in part because prolonged coevolution with one host species limits their ability to evade restriction factors in other species. However, the limitation in host range is incomplete. For example, rhesus CMV (RhCMV) can replicate in human cells, albeit much less efficiently than in rhesus cells. Previously we reported that the protein kinase R (PKR) antagonist encoded by RhCMV, rTRS1, has limited activity against human PKR but is nonetheless necessary and sufficient to enable RhCMV replication in human fibroblasts (HF). We now show that knockout of PKR in human cells or treatment with the eIF2B agonist ISRIB, which overcomes the translational inhibition resulting from PKR activation, augments RhCMV replication in HF, indicating that human PKR contributes to the inefficiency of RhCMV replication in HF. Serial passage of RhCMV in HF reproducibly selected for viruses with improved fitness in human cells. The evolved viruses contain an inverted duplication of the terminal 6.8 kb of the genome, including rTRS1. The duplication replaces ~11.8 kb just downstream of an internal sequence element, pac1-like, which is very similar to the pac1 cleavage and packaging signal found near the terminus of the genome. Plaque-purified evolved viruses produced at least twice as much rTRS1 as the parental RhCMV and blocked the PKR pathway more effectively in HF. Southern blots revealed that unlike the parental RhCMV, viruses with the inverted duplication isomerize in a manner similar to HCMV and other herpesviruses that have internal repeat sequences. The apparent ease with which this duplication event occurs raises the possibility that the pac1-like site, which is conserved in Old World monkey CMV genomes, may serve a function in facilitating rapid adaptation to evolutionary obstacles.

AUTHOR SUMMARYRhesus macaque CMV (RhCMV) is an important model for human CMV (HCMV) pathogenesis and vaccine development. Therefore, it is important to understand the similarities and differences in infectivity and interaction of these viruses with their host species. In contrast to the strict species-specificity of HCMV, RhCMV is able to cross species barriers to replicate in human cells. We know from past work that a component of this broader host range is RhCMVs ability to counteract both the rhesus and human versions of a key antiviral factor. Here we delve further into the mechanisms by which RhCMV can adapt to counteract human cellular defenses. We find that RhCMV appears to be poised to undergo a specific genomic rearrangement that facilitates increased replication efficiency in human cells. Besides providing insights into CMV species-specificity and host barriers to cross-species transmission, this work also provides more generalized clues about viral adaptative mechanisms.
]]></description>
<dc:creator>Child, S. J.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>Geballe, A. P.</dc:creator>
<dc:date>2020-11-06</dc:date>
<dc:identifier>doi:10.1101/2020.11.06.371047</dc:identifier>
<dc:title><![CDATA[Rapid adaptation to human protein kinase R by a unique genomic rearrangement in rhesus cytomegalovirus.]]></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/10.1101/2020.11.09.375329v1?rss=1">
<title>
<![CDATA[
The role of XPB/Ssl2 double-stranded DNA translocase processivity in transcription start-site scanning. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.09.375329v1?rss=1</link>
<description><![CDATA[
The general transcription factor TFIIH contains three ATP-dependent catalytic activities. TFIIH functions in nucleotide excision repair primarily as a DNA helicase and in Pol II transcription initiation as a dsDNA translocase and protein kinase. During initiation, the XPB/Ssl2 subunit of TFIIH couples ATP hydrolysis to dsDNA translocation facilitating promoter opening and the kinase module phosphorylates the C-terminal domain to facilitate the transition to elongation. These functions are conserved between metazoans and yeast; however, yeast TFIIH also drives transcription start-site scanning in which Pol II scans downstream DNA to locate productive start-sites. The ten-subunit holo-TFIIH from S. cerevisiae has a processive dsDNA translocase activity required for scanning and a structural role in scanning has been ascribed to the three-subunit TFIIH kinase module. Here, we assess the dsDNA translocase activity of ten-subunit holo- and core-TFIIH complexes (i.e. seven subunits, lacking the kinase module) from both S. cerevisiae and H. sapiens. We find that neither holo nor core human TFIIH exhibit processive translocation, consistent with the lack of start-site scanning in humans. Furthermore, in contrast to holo-TFIIH, the S. cerevisiae core-TFIIH also lacks processive translocation and its dsDNA-stimulated ATPase activity was reduced ~5-fold to a level comparable to the human complexes, potentially explaining the reported upstream shift in start-site observed in the absence of the S. cerevisiae kinase module. These results suggest that neither human nor S. cerevisiae core-TFIIH can translocate efficiently, and that the S. cerevisiae kinase module functions as a processivity factor to allow for robust transcription start-site scanning.
]]></description>
<dc:creator>Tomko, E. J.</dc:creator>
<dc:creator>Luyties, O.</dc:creator>
<dc:creator>Rimel, J. K.</dc:creator>
<dc:creator>Tsai, C.-L.</dc:creator>
<dc:creator>Fuss, J. O.</dc:creator>
<dc:creator>Fishburn, J.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:creator>Tsutakawa, S. E.</dc:creator>
<dc:creator>Taatjes, D. J.</dc:creator>
<dc:creator>Galburt, E. A.</dc:creator>
<dc:date>2020-11-10</dc:date>
<dc:identifier>doi:10.1101/2020.11.09.375329</dc:identifier>
<dc:title><![CDATA[The role of XPB/Ssl2 double-stranded DNA translocase processivity in transcription start-site scanning.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.10.375881v1?rss=1">
<title>
<![CDATA[
The H3.3K27M oncohistone antagonizes reprogramming in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.10.375881v1?rss=1</link>
<description><![CDATA[
Development proceeds by the activation of genes by transcription factors and the inactivation of others by chromatin-mediated gene silencing. In some cases development can be reversed or redirected by mis-expression of master regulator transcription factors. This must involve the activation of previously silenced genes, and such developmental aberrations are thought to underlie a variety of cancers. Here, we express the wing-specific Vestigial master regulator to reprogram the developing eye, and test the role of silencing in reprogramming using an H3.3K27M oncohistone mutation that dominantly inhibits histone H3K27 trimethylation. We find that expression of the oncohistone blocks eye-to-wing reprogramming. CUT&Tag chromatin profiling of mutant tissues shows that H3K27me3 domains are globally reduced with oncohistone expression, suggesting that previous developmental programs must be silenced for effective transformation. Strikingly, mis-expressed Vg and H3.3K27M synergize to stimulate overgrowth of eye tissue, a phenotype that resembles that of mutations in Polycomb Repressive Complex 1 components. Our results imply that growth dysregulation can result from the simple combination of crippled silencing and transcription factor mis-expression, an effect that may explain the origins of oncohistone-bearing cancers.
]]></description>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2020-11-11</dc:date>
<dc:identifier>doi:10.1101/2020.11.10.375881</dc:identifier>
<dc:title><![CDATA[The H3.3K27M oncohistone antagonizes reprogramming in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.11.378893v1?rss=1">
<title>
<![CDATA[
Glia actively sculpt sensory neurons by controlled phagocytosis to tune animal behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.11.378893v1?rss=1</link>
<description><![CDATA[
Glia in the central nervous system engulf neuron fragments during synapse remodeling and recycling of photoreceptor outer-segments. Whether glia passively clear shed neuronal debris, or actively remove neuron fragments is unknown. How pruning of single-neuron endings impacts animal behavior is also unclear. Here we report that adult C. elegans AMsh glia engulf sensory endings of the AFD thermosensory neuron. Engulfment is regulated by temperature, AFDs sensory input, and tracks AFD activity. Phosphatidylserine (PS) flippase TAT-1/ATP8A, functions with glial PS-receptor PSR-1/PSR and PAT-2/-integrin to initiate engulfment. Glial CED-10/Rac1 GTPase, acting through a conserved GEF complex, executes phagocytosis using the actin-remodeler WSP-1/nWASp and the membrane-sealing factor EFF-1 fusogen. CED-10 levels determine engulfment rates, and engulfment-defective mutants exhibit altered AFD-ending shape and thermosensory behavior. Our findings reveal a molecular pathway underpinning glia-dependent phagocytosis in a peripheral sense-organ, and demonstrate that glia actively engulf neuron-fragments, with profound consequences on neuron shape and animal behavior.
]]></description>
<dc:creator>Raiders, S.</dc:creator>
<dc:creator>Black, E. C.</dc:creator>
<dc:creator>Bae, A.</dc:creator>
<dc:creator>MacFarlane, S.</dc:creator>
<dc:creator>Shaham, S.</dc:creator>
<dc:creator>Singhvi, A.</dc:creator>
<dc:date>2020-11-12</dc:date>
<dc:identifier>doi:10.1101/2020.11.11.378893</dc:identifier>
<dc:title><![CDATA[Glia actively sculpt sensory neurons by controlled phagocytosis to tune animal behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.16.383307v1?rss=1">
<title>
<![CDATA[
Massively parallel phenotyping of variant impact in cancer with Perturb-seq reveals a shift in the spectrum of cell states induced by somatic mutations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.16.383307v1?rss=1</link>
<description><![CDATA[
Genome sequencing studies have identified millions of somatic variants in cancer, but their phenotypic impact remains challenging to predict. Current experimental approaches to distinguish between functionally impactful and neutral variants require customized phenotypic assays that often report on average effects, and are not easily scaled. Here, we develop a generalizable, high-dimensional, and scalable approach to functionally assess variant impact in single cells by pooled Perturb-seq. Specifically, we assessed the impact of 200 TP53 and KRAS variants in >300,000 single lung cancer cells, and used the profiles to categorize variants into phenotypic subsets to distinguish gain-of-function, loss-of-function and dominant negative variants, which we validated by comparison to orthogonal assays. Surprisingly, KRAS variants did not merely fit into discrete functional categories, but rather spanned a continuum of gain-of-function phenotypes driven by quantitative shifts in cell composition at the single cell level. We further discovered novel gain-of-function KRAS variants whose impact could not have been predicted solely by their occurrence in patient samples. Our work provides a scalable, gene-agnostic method for coding variant impact phenotyping, which can be applied in cancer and other diseases driven by somatic or germline coding mutations.
]]></description>
<dc:creator>Ursu, O.</dc:creator>
<dc:creator>Neal, J. T.</dc:creator>
<dc:creator>Shea, E.</dc:creator>
<dc:creator>Thakore, P. I.</dc:creator>
<dc:creator>Jerby-Arnon, L.</dc:creator>
<dc:creator>Nguyen, L.</dc:creator>
<dc:creator>Dionne, D.</dc:creator>
<dc:creator>Diaz, C.</dc:creator>
<dc:creator>Bauman, J.</dc:creator>
<dc:creator>Mosaad, M.</dc:creator>
<dc:creator>Fagre, C.</dc:creator>
<dc:creator>Giacomelli, A.</dc:creator>
<dc:creator>Ly, S. H.</dc:creator>
<dc:creator>Rozenblatt-Rosen, O.</dc:creator>
<dc:creator>Hahn, W.</dc:creator>
<dc:creator>Aguirre, A.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:creator>Regev, A.</dc:creator>
<dc:creator>Boehm, J. S.</dc:creator>
<dc:date>2020-11-17</dc:date>
<dc:identifier>doi:10.1101/2020.11.16.383307</dc:identifier>
<dc:title><![CDATA[Massively parallel phenotyping of variant impact in cancer with Perturb-seq reveals a shift in the spectrum of cell states induced by somatic mutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.16.385278v1?rss=1">
<title>
<![CDATA[
High resolution profiling of pathways of escape for SARS-CoV-2 spike-binding antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.16.385278v1?rss=1</link>
<description><![CDATA[
Defining long-term protective immunity to SARS-CoV-2 is one of the most pressing questions of our time and will require a detailed understanding of potential ways this virus can evolve to escape immune protection. Immune protection will most likely be mediated by antibodies that bind to the viral entry protein, Spike (S). Here we used Phage-DMS, an approach that comprehensively interrogates the effect of all possible mutations on binding to a protein of interest, to define the profile of antibody escape to the SARS-CoV-2 S protein using COVID-19 convalescent plasma. Antibody binding was common in two regions: the fusion peptide and linker region upstream of the heptad repeat region 2. However, escape mutations were variable within these immunodominant regions. There was also individual variation in less commonly targeted epitopes. This study provides a granular view of potential antibody escape pathways and suggests there will be individual variation in antibody-mediated virus evolution.
]]></description>
<dc:creator>Garrett, M. E.</dc:creator>
<dc:creator>Galloway, J.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Itell, H. L.</dc:creator>
<dc:creator>Stoddard, C. I.</dc:creator>
<dc:creator>Wolf, C. R.</dc:creator>
<dc:creator>Logue, J. K.</dc:creator>
<dc:creator>McDonald, D.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2020-11-16</dc:date>
<dc:identifier>doi:10.1101/2020.11.16.385278</dc:identifier>
<dc:title><![CDATA[High resolution profiling of pathways of escape for SARS-CoV-2 spike-binding antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.24.394932v1?rss=1">
<title>
<![CDATA[
A functional module states framework reveals cell states for drug and target prediction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.24.394932v1?rss=1</link>
<description><![CDATA[
Cells are complex systems in which many functions are performed by different genetically-defined and encoded functional modules. To systematically understand how these modules respond to drug or genetic perturbations, we developed a Functional Module States framework. Using this framework, we 1) defined the drug induced transcriptional state space for breast cancer cell lines using large public gene expression datasets, and revealed that the transcriptional states are associated with drug concentration and drug targets; 2) identified potential targetable vulnerabilities through integrative analysis of transcriptional states after drug treatment and gene knockdown associated cancer dependency; and 3) used functional module states to predict transcriptional state-dependent drug sensitivity and built prediction models using the functional module states for drug response. This approach demonstrates a similar prediction performance as do approaches using high dimensional gene expression values, with the added advantage of more clearly revealing biologically relevant transcriptional states and key regulators.
]]></description>
<dc:creator>Qin, G.</dc:creator>
<dc:creator>Knijnenburg, T.</dc:creator>
<dc:creator>Gibbs, D.</dc:creator>
<dc:creator>Moser, R.</dc:creator>
<dc:creator>Monnat, R. J.</dc:creator>
<dc:creator>Kemp, C.</dc:creator>
<dc:creator>Shmulevich, I.</dc:creator>
<dc:date>2020-11-25</dc:date>
<dc:identifier>doi:10.1101/2020.11.24.394932</dc:identifier>
<dc:title><![CDATA[A functional module states framework reveals cell states for drug and target prediction]]></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/10.1101/2020.11.24.396713v1?rss=1">
<title>
<![CDATA[
Chromatin Fiber Folding Represses Transcription and Loop Extrusion in Quiescent Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.24.396713v1?rss=1</link>
<description><![CDATA[
Determining the conformation of chromatin in cells at the nucleosome level and its relationship to cellular processes has been a central challenge in biology. We show that in quiescent yeast, widespread transcriptional repression coincides with the local compaction of chromatin fibers into structures that are less condensed and more heteromorphic than canonical 30-nanometer forms. Acetylation or substitution of H4 tail residues decompacts fibers and leads to global transcriptional de-repression. Fiber decompaction also increases the rate of loop extrusion by condensin. These findings establish a role for H4 tail-dependent local chromatin fiber folding in regulating transcription and loop extrusion in cells. They also demonstrate the physiological relevance of canonical chromatin fiber folding mechanisms even in the absence of regular 30-nanometer structures.
]]></description>
<dc:creator>Swygert, S. G.</dc:creator>
<dc:creator>Lin, D.</dc:creator>
<dc:creator>Portillo-Ledesma, S.</dc:creator>
<dc:creator>Lin, P.-Y.</dc:creator>
<dc:creator>Hunt, D. R.</dc:creator>
<dc:creator>Kao, C.-F.</dc:creator>
<dc:creator>Schlick, T.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:creator>Tsukiyama, T.</dc:creator>
<dc:date>2020-11-24</dc:date>
<dc:identifier>doi:10.1101/2020.11.24.396713</dc:identifier>
<dc:title><![CDATA[Chromatin Fiber Folding Represses Transcription and Loop Extrusion in Quiescent Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.25.398958v1?rss=1">
<title>
<![CDATA[
3D Adaptive Optical Nanoscopy for Thick Specimen Imaging at sub-50 nm Resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.25.398958v1?rss=1</link>
<description><![CDATA[
Understanding cellular organization demands the best possible spatial resolution in all three dimensions (3D). In fluorescence microscopy, this is achieved by 4Pi nanoscopy methods that combine the concepts of using two opposing objectives for optimal diffraction-limited 3D resolution with switching fluorescent molecules between bright and dark states to break the diffraction limit. However, optical aberrations have limited these nanoscopes to thin samples and prevented their application in thick specimens. Here, we have developed a nanoscope that, by utilizing an advanced adaptive optics strategy, achieves sub-50 nm isotropic resolution of structures such as neuronal synapses and ring canals previously inaccessible in tissue.
]]></description>
<dc:creator>Hao, X.</dc:creator>
<dc:creator>Allgeyer, E. S.</dc:creator>
<dc:creator>Antonello, J.</dc:creator>
<dc:creator>Watters, K.</dc:creator>
<dc:creator>Gerdes, J. A.</dc:creator>
<dc:creator>Schroeder, L. K.</dc:creator>
<dc:creator>Bottanelli, F.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Kidd, P.</dc:creator>
<dc:creator>Lessard, M. D.</dc:creator>
<dc:creator>Rothman, J. E.</dc:creator>
<dc:creator>Cooley, L.</dc:creator>
<dc:creator>Biederer, T.</dc:creator>
<dc:creator>Booth, M. J.</dc:creator>
<dc:creator>Bewersdorf, J.</dc:creator>
<dc:date>2020-11-27</dc:date>
<dc:identifier>doi:10.1101/2020.11.25.398958</dc:identifier>
<dc:title><![CDATA[3D Adaptive Optical Nanoscopy for Thick Specimen Imaging at sub-50 nm Resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.30.405472v1?rss=1">
<title>
<![CDATA[
Prospective mapping of viral mutations that escape antibodies used to treat COVID-19 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.30.405472v1?rss=1</link>
<description><![CDATA[
Antibodies are becoming a frontline therapy for SARS-CoV-2, but the risk of viral evolutionary escape remains unclear. Here we map how all mutations to SARS-CoV-2s receptor-binding domain (RBD) affect binding by the antibodies in Regenerons REGN-COV2 cocktail and Eli Lillys LY-CoV016. These complete maps uncover a single amino-acid mutation that fully escapes the REGN-COV2 cocktail, which consists of two antibodies targeting distinct structural epitopes. The maps also identify viral mutations that are selected in a persistently infected patient treated with REGN-COV2, as well as in lab viral escape selections. Finally, the maps reveal that mutations escaping each individual antibody are already present in circulating SARS-CoV-2 strains. Overall, these complete escape maps enable immediate interpretation of the consequences of mutations observed during viral surveillance.
]]></description>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Hannon, W. H.</dc:creator>
<dc:creator>Choudhary, M. C.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Li, J. Z.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-12-01</dc:date>
<dc:identifier>doi:10.1101/2020.11.30.405472</dc:identifier>
<dc:title><![CDATA[Prospective mapping of viral mutations that escape antibodies used to treat COVID-19]]></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/10.1101/2020.12.01.406611v1?rss=1">
<title>
<![CDATA[
Designed proteins assemble antibodies into modular nanocages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.01.406611v1?rss=1</link>
<description><![CDATA[
Antibodies are widely used in biology and medicine, and there has been considerable interest in multivalent antibody formats to increase binding avidity and enhance signaling pathway agonism. However, there are currently no general approaches for forming precisely oriented antibody assemblies with controlled valency. We describe the computational design of two-component nanocages that overcome this limitation by uniting form and function. One structural component is any antibody or Fc fusion and the second is a designed Fc-binding homo-oligomer that drives nanocage assembly. Structures of 8 antibody nanocages determined by electron microscopy spanning dihedral, tetrahedral, octahedral, and icosahedral architectures with 2, 6, 12, and 30 antibodies per nanocage match the corresponding computational models. Antibody nanocages targeting cell-surface receptors enhance signaling compared to free antibodies or Fc-fusions in DR5-mediated apoptosis, Tie2-mediated angiogenesis, CD40 activation, and T cell proliferation; nanocage assembly also increases SARS-CoV-2 pseudovirus neutralization by -SARS-CoV-2 monoclonal antibodies and Fc-ACE2 fusion proteins. We anticipate that the ability to assemble arbitrary antibodies without need for covalent modification into highly ordered assemblies with different geometries and valencies will have broad impact in biology and medicine.
]]></description>
<dc:creator>Divine, R.</dc:creator>
<dc:creator>Dang, H. V.</dc:creator>
<dc:creator>Ueda, G.</dc:creator>
<dc:creator>Fallas, J. A.</dc:creator>
<dc:creator>Vulovic, I.</dc:creator>
<dc:creator>Sheffler, W.</dc:creator>
<dc:creator>Saini, S.</dc:creator>
<dc:creator>Zhao, Y. T.</dc:creator>
<dc:creator>Raj, I. X.</dc:creator>
<dc:creator>Morawski, P. A.</dc:creator>
<dc:creator>Jennewein, M. F.</dc:creator>
<dc:creator>Homad, L. J.</dc:creator>
<dc:creator>Wan, Y.-H.</dc:creator>
<dc:creator>Tooley, M. R.</dc:creator>
<dc:creator>Seeger, F.</dc:creator>
<dc:creator>Fahning, M. L.</dc:creator>
<dc:creator>Etemadi, A.</dc:creator>
<dc:creator>Lazarovits, J.</dc:creator>
<dc:creator>Roederer, A.</dc:creator>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>Stewart, L.</dc:creator>
<dc:creator>Mazloomi, M.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:creator>Campbell, D. J.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Ruohola-Baker, H.</dc:creator>
<dc:creator>Mathieu, J.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2020-12-01</dc:date>
<dc:identifier>doi:10.1101/2020.12.01.406611</dc:identifier>
<dc:title><![CDATA[Designed proteins assemble antibodies into modular nanocages]]></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/10.1101/2020.12.07.415653v1?rss=1">
<title>
<![CDATA[
Evaluating Specimen Quality and Results from a Community-Wide, Home-Based Respiratory Surveillance Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.07.415653v1?rss=1</link>
<description><![CDATA[
IntroductionWhile influenza and other respiratory pathogens cause significant morbidity and mortality, the community-based burden of these infections remains incompletely understood. The development of novel methods to detect respiratory infections is essential for mitigating epidemics and developing pandemic-preparedness infrastructure.

MethodsFrom October 2019 to March 2020, we conducted a home-based cross-sectional study in the greater Seattle area, utilizing electronic consent and data collection instruments. Participants received nasal swab collection kits via rapid delivery within 24 hours of self-reporting respiratory symptoms. Samples were returned to the laboratory and were screened for 26 respiratory pathogens and a human marker. Participant data were recorded via online survey at the time of sample collection and one week later.

ResultsOf the 4,572 consented participants, 4,359 (95.3%) received a home swab kit, and 3,648 (83.7%) returned a nasal specimen for respiratory pathogen screening. The 3,638 testable samples had a mean RNase P CRT value of 19.0 (SD: 3.4) and 1,232 (33.9%) samples had positive results for one or more pathogens, including 645 (17.7%) influenza-positive specimens. Among the testable samples, the median time between shipment of the home swab kit and completion of laboratory testing was 8 days [IQR: 7.0-14.0].

DiscussionHome-based surveillance using online participant enrollment and specimen self-collection is a feasible method for community-level monitoring of influenza and other respiratory pathogens, which can readily be adapted for use during pandemics.
]]></description>
<dc:creator>Kim, A. E.</dc:creator>
<dc:creator>Brandstetter, E.</dc:creator>
<dc:creator>Wilcox, N.</dc:creator>
<dc:creator>Heimonen, J.</dc:creator>
<dc:creator>Graham, C.</dc:creator>
<dc:creator>Han, P. D.</dc:creator>
<dc:creator>Starita, L. M.</dc:creator>
<dc:creator>McCulloch, D. J.</dc:creator>
<dc:creator>Casto, A. M.</dc:creator>
<dc:creator>Nickerson, D. A.</dc:creator>
<dc:creator>Van de Loo, M. M.</dc:creator>
<dc:creator>Mooney, J.</dc:creator>
<dc:creator>Ilcisin, M.</dc:creator>
<dc:creator>Fay, K. A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Sibley, T. R.</dc:creator>
<dc:creator>Lyon, V.</dc:creator>
<dc:creator>Geyer, R. E.</dc:creator>
<dc:creator>Thompson, M.</dc:creator>
<dc:creator>Lutz, B. R.</dc:creator>
<dc:creator>Rieder, M. J.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Boeckh, M.</dc:creator>
<dc:creator>Englund, J. A.</dc:creator>
<dc:creator>Chu, H.</dc:creator>
<dc:date>2020-12-08</dc:date>
<dc:identifier>doi:10.1101/2020.12.07.415653</dc:identifier>
<dc:title><![CDATA[Evaluating Specimen Quality and Results from a Community-Wide, Home-Based Respiratory Surveillance Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.10.420257v1?rss=1">
<title>
<![CDATA[
Integrative genomics identifies lncRNA regulatory networks across 1,044 pediatric leukemias and solid tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.10.420257v1?rss=1</link>
<description><![CDATA[
Long non-coding RNAs (lncRNAs) play an important role in gene regulation and contribute to tumorigenesis. While pan-cancer studies of lncRNA expression have been performed for adult malignancies, the lncRNA landscape across pediatric cancers remains largely uncharted. Here, we curate RNA sequencing data for 1,044 pediatric leukemia and solid tumors and integrate paired tumor whole genome sequencing and epigenetic data in relevant cell line models to explore lncRNA expression, regulation, and association with cancer. We report a total of 2,657 robustly expressed lncRNAs across six pediatric cancers, including 1,142 exhibiting histotype-specific expression. DNA copy number alterations contributed to lncRNA dysregulation at a proportion comparable to protein coding genes. Application of a multi-dimensional framework to identify and prioritize lncRNAs impacting gene networks revealed that lncRNAs dysregulated in pediatric cancer are associated with proliferation, metabolism, and DNA damage hallmarks. Analysis of upstream regulation via cell-type specific transcription factors further implicated distinct histotype-specific and developmental lncRNAs. We integrated our analyses to prioritize lncRNAs for experimental validation and showed that silencing of TBX2-AS1, our top-prioritized neuroblastoma-specific lncRNA, resulted in significant growth inhibition of neuroblastoma cells, confirming our computational predictions. Taken together, these data provide a comprehensive characterization of lncRNA regulation and function in pediatric cancers and pave the way for future mechanistic studies.
]]></description>
<dc:creator>Modi, A.</dc:creator>
<dc:creator>Lopez, G.</dc:creator>
<dc:creator>Conkrite, K. L.</dc:creator>
<dc:creator>Leung, T. C.</dc:creator>
<dc:creator>Ramanan, S.</dc:creator>
<dc:creator>Cheung, D.</dc:creator>
<dc:creator>Su, C.</dc:creator>
<dc:creator>Manduchi, E.</dc:creator>
<dc:creator>Johnson, M. E.</dc:creator>
<dc:creator>Gadd, S.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Smith, M. A.</dc:creator>
<dc:creator>Guidry Auvil, J. M.</dc:creator>
<dc:creator>Gerhard, D. S.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Perlman, E. J.</dc:creator>
<dc:creator>Hunger, S. P.</dc:creator>
<dc:creator>Maris, J. M.</dc:creator>
<dc:creator>Wells, A. D.</dc:creator>
<dc:creator>Grant, S. F. A.</dc:creator>
<dc:creator>Diskin, S. J.</dc:creator>
<dc:date>2020-12-11</dc:date>
<dc:identifier>doi:10.1101/2020.12.10.420257</dc:identifier>
<dc:title><![CDATA[Integrative genomics identifies lncRNA regulatory networks across 1,044 pediatric leukemias and solid tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.11.418129v1?rss=1">
<title>
<![CDATA[
A viral histone-like protein exploits H1-HMGB1 antagonism for chromatin invasion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.11.418129v1?rss=1</link>
<description><![CDATA[
Virus infection necessarily requires redirecting cellular resources towards viral progeny production. Adenovirus encodes the histone-like protein VII that causes catastrophic global reorganization of host chromatin to promote virus infection. Protein VII recruits the family of high mobility group box (HMGB) proteins to chromatin along with the histone chaperone SET. As a consequence of this recruitment, we find that protein VII causes chromatin-depletion of several linker histone H1 isoforms. The relationship between linker histone H1 and the functionally opposite HMGB proteins is critical for higher order chromatin structure. However, the physiological consequences of perturbing this relationship are largely unknown. Here, we employ complementary systems in Saccharomyces cerevisiae and human cells to demonstrate that adenovirus protein VII disrupts the H1-HMGB balance to obstruct the cell cycle. We find that protein VII causes an accumulation of G2/M cells both in yeast and human systems, underscoring the high conservation of this chromatin vulnerability. In contrast, adenovirus E1A and E1B proteins are well-established to override cell cycle regulation and promote transformation of human cells. Strikingly, we find that protein VII obstructs the cell cycle even in the presence of E1A and E1B, suggesting that protein VII-directed cell cycle disruption ensures host resources are directed towards viral proliferation during infection. Together, our results demonstrate that protein VII targets H1-HMGB1 antagonism to obstruct cell cycle progression, revealing an unexpected chromatin vulnerability exploited for viral benefit.
]]></description>
<dc:creator>Lynch, K. L.</dc:creator>
<dc:creator>Bat-Erdene, M.</dc:creator>
<dc:creator>Dillon, M. R.</dc:creator>
<dc:creator>Lewis, H. C.</dc:creator>
<dc:creator>Avgousti, D. C.</dc:creator>
<dc:date>2020-12-11</dc:date>
<dc:identifier>doi:10.1101/2020.12.11.418129</dc:identifier>
<dc:title><![CDATA[A viral histone-like protein exploits H1-HMGB1 antagonism for chromatin invasion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.17.423211v1?rss=1">
<title>
<![CDATA[
Identification of Celastrol as a novel HIV-1 Latency  Reversal Agent by an Image-Based Screen 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.17.423211v1?rss=1</link>
<description><![CDATA[
Although current antiretroviral therapies (ART) are successful in controlling HIV-1 infection, a stable viral reservoir reactivates when ART is discontinued. Consequently, there is a major research effort to develop approaches to disrupt the latent viral reservoir and enhance the immune systems ability to clear HIV-1. A number of small molecules, termed latency reversal agents (LRAs), have been identified which can reactivate latent HIV-1 in cell lines and patients cells ex vivo. However, clinical trials have suggested that combinations of LRAs will be required to efficiently reactivate HIV-1 in vivo, especially LRAs that act synergistically by functioning through distinct pathways. To identify novel LRAs, we used an image-based assay to screen a natural compound library for the ability to induce a low level of aggregation of resting primary CD4+ T cells from healthy donors. We identified celastrol as a novel LRA. Celastrol functions synergistically with other classes of LRA to reactivate latent HIV-1 in a Jurkat cell line, suggesting a novel mechanism in its LRA activity. Additionally, celastrol does not appear to activate resting CD4+ T cells at levels at which it can reactivate latent HIV-1. Celastrol appears to represent a novel class of LRAs and it therefore can serve as a lead compound for LRA development.
]]></description>
<dc:creator>Rice, A. P.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Hu, P.-W. P.</dc:creator>
<dc:creator>Dubrulle, J. P.</dc:creator>
<dc:creator>Stossi, F.</dc:creator>
<dc:creator>Mancini, M.</dc:creator>
<dc:creator>Nikolai, B.</dc:creator>
<dc:date>2020-12-17</dc:date>
<dc:identifier>doi:10.1101/2020.12.17.423211</dc:identifier>
<dc:title><![CDATA[Identification of Celastrol as a novel HIV-1 Latency  Reversal Agent by an Image-Based Screen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.17.423313v1?rss=1">
<title>
<![CDATA[
A human coronavirus evolves antigenically to escape antibody immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.17.423313v1?rss=1</link>
<description><![CDATA[
There is intense interest in antibody immunity to coronaviruses. However, it is unknown if coronaviruses evolve to escape such immunity, and if so, how rapidly. Here we address this question by characterizing the historical evolution of human coronavirus 229E. We identify human sera from the 1980s and 1990s that have neutralizing titers against contemporaneous 229E that are comparable to the anti-SARS-CoV-2 titers induced by SARS-CoV-2 infection or vaccination. We test these sera against 229E strains isolated after sera collection, and find that neutralizing titers are lower against these "future" viruses. In some cases, sera that neutralize contemporaneous 229E viral strains with titers >1:100 do not detectably neutralize strains isolated 8-17 years later. The decreased neutralization of "future" viruses is due to antigenic evolution of the viral spike, especially in the receptor-binding domain. If these results extrapolate to other coronaviruses, then it may be advisable to periodically update SARS-CoV-2 vaccines.
]]></description>
<dc:creator>Eguia, R.</dc:creator>
<dc:creator>Crawford, K. H. D.</dc:creator>
<dc:creator>Stevens-Ayers, T.</dc:creator>
<dc:creator>Kelnhofer-Millevolte, L.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>Englund, J. A.</dc:creator>
<dc:creator>Boeckh, M. J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2020-12-18</dc:date>
<dc:identifier>doi:10.1101/2020.12.17.423313</dc:identifier>
<dc:title><![CDATA[A human coronavirus evolves antigenically to escape antibody immunity]]></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/10.1101/2020.12.20.423710v1?rss=1">
<title>
<![CDATA[
Discovery of synthetic lethal and tumor suppressive paralog pairs in the human genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.20.423710v1?rss=1</link>
<description><![CDATA[
CRISPR knockout screens have accelerated the discovery of important cancer genetic dependencies. However, traditional CRISPR-Cas9 screens are limited in their ability to assay the function of redundant or duplicated genes. Paralogs in multi-gene families constitute two-thirds of the protein-coding genome, so this blind spot is the rule, not the exception. To overcome the limitations of single gene CRISPR knockout screens, we developed paired guide RNAs for Paralog gENetic interaction mapping (pgPEN), a pooled CRISPR/Cas9 approach which targets over a thousand duplicated human paralogs in single knockout and double knockout configurations. We applied pgPEN to two cell lineages and discovered that over 10% of human paralogs exhibit synthetic lethality in at least one cellular context. We recovered known synthetic lethal paralogs such as MAP2K1/MAP2K2, important drug targets such as CDK4/CDK6, and numerous other synthetic lethal pairs such as CCNL1/CCNL2. In addition, we identified ten tumor suppressive paralog pairs whose compound loss promotes cell growth. These findings identify a large number of previously unidentified essential gene families and nominate new druggable targets for oncology drug discovery.

HighlightsO_LIComprehensive genetic interaction mapping of 1,030 human duplicated paralogs using a dual targeting CRISPR/Cas9 approach
C_LIO_LIDuplicated paralogs are highly enriched for genetic interactions
C_LIO_LISynthetic lethal paralogs include CCNL1/CCNL2, CDK4/CDK6, and GSK3A/GSK3B
C_LIO_LITumor suppressor paralog pairs include CDKN2A/CDKN2B and FBXO25/FBXO32
C_LI
]]></description>
<dc:creator>Parrish, P. C. R.</dc:creator>
<dc:creator>Thomas, J. D.</dc:creator>
<dc:creator>Kamlapurkar, S.</dc:creator>
<dc:creator>Gabel, A.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:creator>Berger, A. H.</dc:creator>
<dc:date>2020-12-21</dc:date>
<dc:identifier>doi:10.1101/2020.12.20.423710</dc:identifier>
<dc:title><![CDATA[Discovery of synthetic lethal and tumor suppressive paralog pairs in the human genome]]></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/10.1101/2020.12.21.423274v1?rss=1">
<title>
<![CDATA[
Multistability and consequent phenotypic plasticity in AMPK-Akt double negative feedback loop in cancer cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.21.423274v1?rss=1</link>
<description><![CDATA[
Adaptation and survival of cancer cells to various stress and growth factor conditions is crucial for successful metastasis. A double-negative feedback loop between two serine/threonine kinases AMPK and Akt can regulate the adaptation of breast cancer cells to matrix-deprivation stress. This feedback loop can generate majorly two phenotypes or cell states: matrix detachment-triggered pAMPKhigh/ pAktlow state, and matrix (re)attachment-triggered pAkthigh/ pAMPKlow state. However, whether these two cell states can exhibit phenotypic plasticity and heterogeneity in a given cell population, i.e., whether they can co-exist and undergo spontaneous switching to generate the other subpopulation, remains unclear. Here, we develop a mechanism-based mathematical model that captures the set of experimentally reported interactions among AMPK and Akt. Our simulations suggest that the AMPK-Akt feedback loop can give rise to two co-existing phenotypes (pAkthigh/ pAMPKlow and pAMPKhigh/pAktlow) in specific parameter regimes. Next, to test the model predictions, we segregated these two subpopulations in MDA-MB-231 cells and observed that each of them was capable of switching to another in adherent conditions. Finally, the predicted trends are supported by clinical data analysis of TCGA breast cancer and pan-cancer cohorts that revealed negatively correlated pAMPK and pAkt protein levels. Overall, our integrated computational-experimental approach unravels that AMPK-Akt feedback loop can generate multistability and drive phenotypic switching and heterogeneity in a cancer cell population.
]]></description>
<dc:creator>Chedere, A.</dc:creator>
<dc:creator>Hari, K.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Rangarajan, A.</dc:creator>
<dc:creator>Jolly, M. K.</dc:creator>
<dc:date>2020-12-22</dc:date>
<dc:identifier>doi:10.1101/2020.12.21.423274</dc:identifier>
<dc:title><![CDATA[Multistability and consequent phenotypic plasticity in AMPK-Akt double negative feedback loop in cancer 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/10.1101/2020.12.22.423783v1?rss=1">
<title>
<![CDATA[
Rare coding variants in 35 genes associate with circulating lipid levels: a multi-ancestry analysis of 170,000 exomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.22.423783v1?rss=1</link>
<description><![CDATA[
Large-scale gene sequencing studies for complex traits have the potential to identify causal genes with therapeutic implications. We performed gene-based association testing of blood lipid levels with rare (minor allele frequency<1%) predicted damaging coding variation using sequence data from >170,000 individuals from multiple ancestries: 97,493 European, 30,025 South Asian, 16,507 African, 16,440 Hispanic/Latino, 10,420 East Asian, and 1,182 Samoan. We identified 35 genes associated with circulating lipid levels. Ten of these: ALB, SRSF2, JAK2, CREB3L3, TMEM136, VARS, NR1H3, PLA2G12A, PPARG and STAB1 have not been implicated for lipid levels using rare coding variation in population-based samples. We prioritize 32 genes identified in array-based genome-wide association study (GWAS) loci based on gene-based associations, of which three: EVI5, SH2B3, and PLIN1, had no prior evidence of rare coding variant associations. Most of the associated genes showed evidence of association in multiple ancestries. Also, we observed an enrichment of gene-based associations for low-density lipoprotein cholesterol drug target genes, and for genes closest to GWAS index single nucleotide polymorphisms (SNP). Our results demonstrate that gene-based associations can be beneficial for drug target development and provide evidence that the gene closest to the array-based GWAS index SNP is often the functional gene for blood lipid levels.
]]></description>
<dc:creator>Hindy, G.</dc:creator>
<dc:creator>Dornbos, P.</dc:creator>
<dc:creator>Chaffin, M. D.</dc:creator>
<dc:creator>Liu, D. J.</dc:creator>
<dc:creator>Wang, M. X.</dc:creator>
<dc:creator>Aguilar-Salinas, C. A.</dc:creator>
<dc:creator>Antonacci-Fulton, L.</dc:creator>
<dc:creator>Ardissino, D.</dc:creator>
<dc:creator>Arnett, D. K.</dc:creator>
<dc:creator>Aslibekyan, S.</dc:creator>
<dc:creator>Atzmon, G.</dc:creator>
<dc:creator>Ballantyne, C. M.</dc:creator>
<dc:creator>Barajas-Olmos, F.</dc:creator>
<dc:creator>Barzilai, N.</dc:creator>
<dc:creator>Becker, L. C.</dc:creator>
<dc:creator>Bielak, L. F.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Bonnycastle, L. L.</dc:creator>
<dc:creator>Bottinger, E.</dc:creator>
<dc:creator>Bowden, D. W.</dc:creator>
<dc:creator>Bown, M. J.</dc:creator>
<dc:creator>Brody, J. A.</dc:creator>
<dc:creator>Broome, J. G.</dc:creator>
<dc:creator>Burtt, N. P.</dc:creator>
<dc:creator>Cade, B. E.</dc:creator>
<dc:creator>Centeno-Cruz, F.</dc:creator>
<dc:creator>Chan, E.</dc:creator>
<dc:creator>Chang, Y.-C.</dc:creator>
<dc:creator>Chen, Y.-D. I.</dc:creator>
<dc:creator>Cheng, C.-Y.</dc:creator>
<dc:creator>Choi, W. J.</dc:creator>
<dc:creator>Chowdhury, R.</dc:creator>
<dc:creator>Contreras-Cubas, C.</dc:creator>
<dc:creator>Cordova, E. J.</dc:creator>
<dc:creator>Correa, A.</dc:creator>
<dc:creator>Cupples, L. A.</dc:creator>
<dc:creator>Curran, J. E.</dc:creator>
<dc:creator>Danesh,</dc:creator>
<dc:date>2020-12-23</dc:date>
<dc:identifier>doi:10.1101/2020.12.22.423783</dc:identifier>
<dc:title><![CDATA[Rare coding variants in 35 genes associate with circulating lipid levels: a multi-ancestry analysis of 170,000 exomes]]></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/10.1101/2020.12.23.424063v1?rss=1">
<title>
<![CDATA[
MLX balances metabolism and stress to suppress apoptosis and maintain spermatogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.23.424063v1?rss=1</link>
<description><![CDATA[
Male germ cell production is a metabolically-driven and apoptosis-prone process. Here we show that the glucose-sensing transcription factor MLX, and its binding partner MondoA, are both required for male fertility in the mouse, as well as survival of human tumor cells derived from the male germ line. Loss of Mlx results in altered metabolism as well as activation of multiple stress pathways and germ cell apoptosis in the testes. This is concomitant with dysregulation of the expression of male-specific germ cell transcripts and proteins, manifesting as oligoasthenoteratozoospermia (OAT). Our genomic and functional analyses identify loci directly bound by MLX involved in these processes, including metabolic targets, apoptotic effectors and obligate components of male-specific germ cell development. These in vivo and in vitro studies implicate MLX and other members of the proximal MYC network, such as MNT, in regulation of metabolism and differentiation, as well as in suppression of intrinsic and extrinsic death signaling pathways in both spermatogenesis and male germ cell tumors.

Highlights- The MAX-like bHLHLZ protein MLX is required for male fertility, but not embryonic development.
- MLX and its heterodimeric partner MondoA are each required for both male fertility and survival of male germ cell tumors.
- Genomic analysis identifies direct MLX targets associated with metabolism, stress and male germ cell development.
- Loss of MLX alters MYC network genome occupancy and transcriptional output.
]]></description>
<dc:creator>Carroll, P.</dc:creator>
<dc:creator>Cheng, P.-F.</dc:creator>
<dc:creator>Freie, B. W.</dc:creator>
<dc:creator>Kasinathan, S.</dc:creator>
<dc:creator>Gu, H.</dc:creator>
<dc:creator>Hedrich, T.</dc:creator>
<dc:creator>Dowdle, J. A.</dc:creator>
<dc:creator>Venkataramani, V.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Raftery, D.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Ayer, D. E.</dc:creator>
<dc:creator>Muller, C. H.</dc:creator>
<dc:creator>Eisenman, R. N. N.</dc:creator>
<dc:date>2020-12-23</dc:date>
<dc:identifier>doi:10.1101/2020.12.23.424063</dc:identifier>
<dc:title><![CDATA[MLX balances metabolism and stress to suppress apoptosis and maintain spermatogenesis]]></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/10.1101/2020.12.24.424260v1?rss=1">
<title>
<![CDATA[
TCR meta-clonotypes for biomarker discovery with tcrdist3: quantification of public, HLA-restricted TCR biomarkers of SARS-CoV-2 infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.24.424260v1?rss=1</link>
<description><![CDATA[
As the mechanistic basis of adaptive cellular antigen recognition, T cell receptors (TCRs) encode clinically valuable information that reflects prior antigen exposure and potential future response. However, despite advances in deep repertoire sequencing, enormous TCR diversity complicates the use of TCR clonotypes as clinical biomarkers. We propose a new framework that leverages antigen-enriched repertoires to form meta-clonotypes - groups of biochemically similar TCRs - that can be used to robustly identify and quantify functionally similar TCRs in bulk repertoires. We apply the framework to TCR data from COVID-19 patients, generating 1831 public TCR meta-clonotypes from the 17 SARS-CoV-2 antigen-enriched repertoires with the strongest evidence of HLA-restriction. Applied to independent cohorts, meta-clonotypes targeting these specific epitopes were more frequently detected in bulk repertoires compared to exact amino acid matches, and 59.7% (1093/1831) were more abundant among COVID-19 patients that expressed the putative restricting HLA allele (FDR < 0.01), demonstrating the potential utility of meta-clonotypes as antigen-specific features for biomarker development. To enable further applications, we developed an open-source software package, tcrdist3, that implements this framework and facilitates flexible workflows for distance-based TCR repertoire analysis.
]]></description>
<dc:creator>Mayer-Blackwell, K.</dc:creator>
<dc:creator>Schattgen, S.</dc:creator>
<dc:creator>Cohen-Lavi, L.</dc:creator>
<dc:creator>Crawford, J. C.</dc:creator>
<dc:creator>Souquette, A.</dc:creator>
<dc:creator>Gaevert, J. A.</dc:creator>
<dc:creator>Hertz, T.</dc:creator>
<dc:creator>Thomas, P. G.</dc:creator>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:creator>Fiore-Gartland, A.</dc:creator>
<dc:date>2020-12-26</dc:date>
<dc:identifier>doi:10.1101/2020.12.24.424260</dc:identifier>
<dc:title><![CDATA[TCR meta-clonotypes for biomarker discovery with tcrdist3: quantification of public, HLA-restricted TCR biomarkers of SARS-CoV-2 infection]]></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/10.1101/2020.12.31.425021v1?rss=1">
<title>
<![CDATA[
Comprehensive mapping of mutations to the SARS-CoV-2 receptor-binding domain that affect recognition by polyclonal human serum antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.31.425021v1?rss=1</link>
<description><![CDATA[
The evolution of SARS-CoV-2 could impair recognition of the virus by human antibody-mediated immunity. To facilitate prospective surveillance for such evolution, we map how convalescent serum antibodies are impacted by all mutations to the spikes receptor-binding domain (RBD), the main target of serum neutralizing activity. Binding by polyclonal serum antibodies is affected by mutations in three main epitopes in the RBD, but there is substantial variation in the impact of mutations both among individuals and within the same individual over time. Despite this inter- and intra-person heterogeneity, the mutations that most reduce antibody binding usually occur at just a few sites in the RBDs receptor binding motif. The most important site is E484, where neutralization by some sera is reduced >10-fold by several mutations, including one in emerging viral lineages in South Africa and Brazil. Going forward, these serum escape maps can inform surveillance of SARS-CoV-2 evolution.
]]></description>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Malone, K. D.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-01-04</dc:date>
<dc:identifier>doi:10.1101/2020.12.31.425021</dc:identifier>
<dc:title><![CDATA[Comprehensive mapping of mutations to the SARS-CoV-2 receptor-binding domain that affect recognition by polyclonal human serum antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.06.425610v1?rss=1">
<title>
<![CDATA[
Coordination of phage genome degradation versus host genome protection by a bifunctional restriction-modification enzyme visualized by CryoEM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.06.425610v1?rss=1</link>
<description><![CDATA[
Restriction enzymes that combine DNA methylation and cleavage activities into a single polypeptide or protein assemblage and that modify just one DNA strand for host protection are capable of more efficient adaptation towards novel target sites. However, they must solve the problem of discrimination between newly replicated and unmodified host sites (needing methylation) and invasive foreign site (needing to lead to cleavage). One solution to this problem might be that the activity that occurs at any given site is dictated by the oligomeric state of the bound enzyme. Methylation requires just a single bound site and is relatively slow, while cleavage requires that multiple unmethylated target sites (often found in incoming, foreign DNA) be brought together into an enzyme-DNA complex to license rapid cleavage. To validate and visualize the basis for such a mechanism, we have determined the catalytic behavior of a bifunctional Type IIL restriction-modification ( RM) enzyme (DrdV) and determined its high-resolution structure at several different stages of assembly and coordination with multiple bound DNA targets using CryoEM. The structures demonstrate a mechanism of cleavage by which an initial dimer is formed between two DNA-bound enzyme molecules, positioning the single endonuclease domain from each enzyme against the others DNA and requiring further oligomerization through differing protein-protein contacts of additional DNA-bound enzyme molecules to enable cleavage. The analysis explains how endonuclease activity is licensed by the presence of multiple target-containing DNA duplexes and provides a clear view of the assembly through 3D space of a DNA-bound RM enzyme  synapse that leads to rapid cleavage of foreign DNA.
]]></description>
<dc:creator>Shen, B. W.</dc:creator>
<dc:creator>Quispe, J. D.</dc:creator>
<dc:creator>Luyten, Y.</dc:creator>
<dc:creator>McGeough, B. C.</dc:creator>
<dc:creator>Morgan, R. D.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:date>2021-01-06</dc:date>
<dc:identifier>doi:10.1101/2021.01.06.425610</dc:identifier>
<dc:title><![CDATA[Coordination of phage genome degradation versus host genome protection by a bifunctional restriction-modification enzyme visualized by CryoEM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.06.425613v1?rss=1">
<title>
<![CDATA[
Extracellular Matrix Dysfunction in Sorsby Patient-Derived Retinal Pigment Epithelium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.06.425613v1?rss=1</link>
<description><![CDATA[
Sorsby Fundus Dystrophy (SFD) is a rare form of macular degeneration that is clinically similar to age-related macular degeneration (AMD), and a histologic hallmark of SFD is a thick layer of extracellular deposits beneath the retinal pigment epithelium (RPE). Previous studies of SFD patient-induced pluripotent stem cell (iPSC) derived RPE differ as to whether these cultures recapitulate this key clinical feature by forming increased drusenoid deposits. The primary purpose of this study is to examine whether SFD patient-derived iPSC-RPE form basal deposits similar to what is found in affected family member SFD globes and to determine whether SFD iPSC RPE may be more oxidatively stressed. We performed a careful comparison of iPSC RPE from three control individuals, multiple iPSC clones from two SFD patients iPSC RPE, and post-mortem eyes of affected SFD family members. We also examined the effect of CRISPR-Cas9 gene correction of the S204C TIMP3 mutation on RPE phenotype. Finally, targeted metabolomics analysis with liquid chromatography and mass spectrometry analysis and stable isotope-labeled metabolite analysis was performed to determine whether SFD RPE are more oxidatively stressed. We found that SFD iPSC-RPE formed significantly more sub-RPE deposits ([~]6-90 m in height) compared to control RPE at 8 weeks. These deposits were similar in composition to the basal laminar drusen found in SFD family member globes by immunofluorescence staining and TEM imaging. S204C TIMP3 correction by CRISPR-Cas9 gene editing in SFD iPSC RPE cells resulted in significantly reduced basal laminar and sub-RPE calcium deposits. We detected a [~]18-fold increase in TIMP3 accumulation in the extracellular matrix (ECM) of SFD RPE, and targeted metabolomics showed that intracellular 4-hydroxyproline, a major breakdown product of collagen, is significantly elevated in SFD RPE, suggesting increased ECM turnover. Finally, SFD RPE cells have decreased intracellular reduced glutathione and were found to be more vulnerable to oxidative stress. Our findings suggest that elements of SFD pathology can be demonstrated in culture which may lead to insights into disease mechanisms.
]]></description>
<dc:creator>Engel, A. L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Khuu, T.</dc:creator>
<dc:creator>Worrall, E.</dc:creator>
<dc:creator>Manson, M. A.</dc:creator>
<dc:creator>Knight, K.</dc:creator>
<dc:creator>Yanagida, A.</dc:creator>
<dc:creator>Qi, J. H.</dc:creator>
<dc:creator>Ramakrishnan, A.</dc:creator>
<dc:creator>Weleber, R. G.</dc:creator>
<dc:creator>Klein, M.</dc:creator>
<dc:creator>Wilson, D. J.</dc:creator>
<dc:creator>Anand-Apte, B.</dc:creator>
<dc:creator>Hurley, J. B.</dc:creator>
<dc:creator>Du, J. L.</dc:creator>
<dc:creator>Chao, J. R.</dc:creator>
<dc:date>2021-01-07</dc:date>
<dc:identifier>doi:10.1101/2021.01.06.425613</dc:identifier>
<dc:title><![CDATA[Extracellular Matrix Dysfunction in Sorsby Patient-Derived Retinal Pigment Epithelium]]></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/10.1101/2021.01.06.425642v1?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/10.1101/2021.01.08.425928v1?rss=1">
<title>
<![CDATA[
Distinct spermiogenic phenotypes underlie sperm elimination in the Segregation Distorter meiotic drive system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.08.425928v1?rss=1</link>
<description><![CDATA[
Segregation Distorter (SD) is a male meiotic drive system in Drosophila melanogaster. Males heterozygous for a selfish SD chromosome rarely transmit the homologous SD+ chromosome. It is well established that distortion results from an interaction between Sd, the primary distorting locus on the SD chromosome and its target, a satellite DNA called Rsp, on the SD+ chromosome. However, the molecular and cellular mechanisms leading to post-meiotic SD+ sperm elimination remain unclear. Here we show that SD/SD+ males of different genotypes but with similarly strong degrees of distortion have distinct spermiogenic phenotypes. In some genotypes, SD+ spermatids fail to fully incorporate protamines after the removal of histones, and degenerate during the individualization stage of spermiogenesis. In contrast, in other SD/SD+ genotypes, protamine incorporation appears less disturbed, yet spermatid nuclei are abnormally compacted, and mature sperm nuclei are eventually released in the seminal vesicle. Our analyses of different SD+ chromosomes suggest that the severity of the spermiogenic defects associates with the copy number of the Rsp satellite. We propose that when Rsp copy number is very high (> 2000), spermatid nuclear compaction defects reach a threshold that triggers a checkpoint controlling sperm chromatin quality to eliminate abnormal spermatids during individualization.
]]></description>
<dc:creator>Herbette, M.</dc:creator>
<dc:creator>Wei, X.</dc:creator>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Larracuente, A. M.</dc:creator>
<dc:creator>Loppin, B.</dc:creator>
<dc:creator>Dubruille, R.</dc:creator>
<dc:date>2021-01-09</dc:date>
<dc:identifier>doi:10.1101/2021.01.08.425928</dc:identifier>
<dc:title><![CDATA[Distinct spermiogenic phenotypes underlie sperm elimination in the Segregation Distorter meiotic drive system]]></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/10.1101/2021.01.08.425930v1?rss=1">
<title>
<![CDATA[
Cysteine is a limiting factor for glioma proliferation and survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.08.425930v1?rss=1</link>
<description><![CDATA[
Nutritional intervention is becoming more prevalent as adjuvant therapy for many cancers in view of the tumor dependence on external sources for some nutrients. We report the dependence of glioma cells on exogenous cysteine/cystine, despite this amino acid being nonessential. 13C-tracing and the analysis of cystathionine synthase and cystathioninase levels revealed the metabolic landscape attributable to cysteine deprivation, and the disconnection between the methionine cycle and the transsulfuration pathway. Therefore, we explored the nutritional deprivation in a mouse model of glioma. Animals subjected to a cysteine/cystine-free diet survived longer, with concomitant reductions in glutathione and cysteine plasma levels. At the end point, however, tumors displayed the ability to synthesize glutathione, although higher levels of oxidative stress were detected. We observed a compensation from the nutritional intervention revealed as the recovery of cysteine-related metabolites in plasma. Our study highlights a time window where cysteine deprivation can be exploited for additional therapeutic strategies.
]]></description>
<dc:creator>Larion, M.</dc:creator>
<dc:creator>Ruiz-Rodado, V.</dc:creator>
<dc:creator>Dowdy, T.</dc:creator>
<dc:creator>Lita, A.</dc:creator>
<dc:creator>Kramp, T.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Jung, J.</dc:creator>
<dc:creator>Dios-Esponera, A.</dc:creator>
<dc:creator>Herold-Mende, C. C.</dc:creator>
<dc:creator>Camphausen, K.</dc:creator>
<dc:creator>Gilbert, M. R.</dc:creator>
<dc:date>2021-01-08</dc:date>
<dc:identifier>doi:10.1101/2021.01.08.425930</dc:identifier>
<dc:title><![CDATA[Cysteine is a limiting factor for glioma proliferation and survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.09.425923v1?rss=1">
<title>
<![CDATA[
Targeting glucose metabolism sensitizes pancreatic cancer to MEK inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.09.425923v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDAC) is almost universally lethal. A critical unmet need exists to explore essential susceptibilities in PDAC and identify druggable targets for tumor maintenance. This is especially challenging in the context of PDAC, in which activating mutations of KRAS oncogene (KRAS*) dominate the genetic landscape. By using an inducible KrasG12D-driven p53 deficient PDAC mouse model (iKras model), we demonstrate that RAF-MEK-MAPK signaling is the major effector for oncogenic Kras-mediated tumor maintenance. However, MEK inhibition has minimal therapeutic effect as single agent for PDAC both in vitro and in vivo. Although MEK inhibition partially downregulates the transcription of glycolysis genes, it surprisingly fails to suppress the glycolysis flux in PDAC cell, which is a major metabolism effector of oncogenic KRAS. Accordingly, In vivo genetic screen identified multiple glycolysis genes as potential targets that may sensitize tumor cells to MAPK inhibition. Furthermore, inhibition of glucose metabolism with low dose 2-deoxyglucose (2DG) in combination with MEK inhibitor dramatically induces apoptosis in KrasG12D-driven PDAC cell in vitro, inhibits xenograft tumor growth and prolongs the overall survival of genetically engineered mouse PDAC model. Molecular and metabolism analyses indicate that co-targeting glycolysis and MAPK signaling results in apoptosis via induction of lethal ER stress. Together, our work suggests that combinatory inhibition of glycolysis and MAPK pathway may serve as an alternative approach to target KRAS-driven PDAC.
]]></description>
<dc:creator>Yan, L.</dc:creator>
<dc:creator>Tu, B.</dc:creator>
<dc:creator>Yao, J.</dc:creator>
<dc:creator>Gong, J.</dc:creator>
<dc:creator>Carugo, A.</dc:creator>
<dc:creator>Bristow, C.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Zhu, C.</dc:creator>
<dc:creator>Dai, B.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Han, L.</dc:creator>
<dc:creator>Feng, N.</dc:creator>
<dc:creator>Jin, Y.</dc:creator>
<dc:creator>Fleming, J.</dc:creator>
<dc:creator>Heffernan, T.</dc:creator>
<dc:creator>Yao, W.</dc:creator>
<dc:creator>Ying, H.</dc:creator>
<dc:date>2021-01-11</dc:date>
<dc:identifier>doi:10.1101/2021.01.09.425923</dc:identifier>
<dc:title><![CDATA[Targeting glucose metabolism sensitizes pancreatic cancer to MEK inhibition]]></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/10.1101/2021.01.11.426199v1?rss=1">
<title>
<![CDATA[
Sustained IL-15 response signature predicts RhCMV/SIV vaccine efficacy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.11.426199v1?rss=1</link>
<description><![CDATA[
Simian immunodeficiency virus (SIV) challenge of rhesus macaques (RMs) vaccinated with Rhesus Cytomegalovirus (RhCMV) vectors expressing SIV proteins (RhCMV/SIV) results in a binary outcome: stringent control and subsequent clearance of highly pathogenic SIV in ~55% of vaccinated RMs with no protection in the remaining 45%. Although previous work suggests that unconventionally restricted, SIV-specific, effector-memory (EM)-biased CD8+ T cell responses are necessary for efficacy, the magnitude of these responses does not predict efficacy, and the basis of protection vs. non-protection in RhCMV/SIV vector-vaccinated RMs has not been elucidated. Here, we report that RhCMV/SIV vector administration strikingly alters the whole blood transcriptome of vaccinated RMs, with the sustained induction of specific immune-related pathways, including non-canonical T cell receptor (TCR), toll-lie receptor (TLR), inflammasome/cell death, and interleukin-15 (IL-15) signaling, significantly predicting protection. The IL-15 gene expression signature was further evaluated in an independent RM IL-15 treatment cohort, revealing that in whole blood the response to IL-15 is inclusive of innate and adaptive immune gene expression networks that link with RhCMV/SIV vaccine efficacy. We also show that this IL-15 response signature similarly tracks with vaccine protection in an independent RhCMV/SIV vaccination/SIV challenge RM validation cohort. Thus, the RhCMV/SIV vaccine imparts a coordinated and persistent induction of innate and adaptive immune pathways featuring IL-15, a known regulator of CD8+ T cell function, that enable vaccine-elicited CD8+ T cells to mediate protection against highly pathogenic SIV challenge.

Author SummarySIV insert-expressing vaccine vectors based on strain 68-1 RhCMV elicit robust, highly effector-memory-biased T cell responses that are associated with an unprecedented level of SIV control after challenge (replication arrest leading to clearance) in slightly over half of vaccinated monkeys. Since efficacy is not predicted by standard measures of immunogenicity, we used functional genomics analysis of RhCMV/SIV vaccinated monkeys with known challenge outcomes to identify immune correlates of protection. We found that arrest of viral replication after challenge significantly correlates with a vaccine-induced response to IL-15 that includes modulation of T cell, inflammation, TLR signaling, and cell death programming. These data suggest that RhCMV/SIV efficacy is not based on chance, but rather, results from a coordinated and sustained vaccine-mediated induction of innate and adaptive immune pathways featuring IL-15, a known regulator of CD8+ effector-memory T cell function, that enable vaccine-elicited CD8+ T cells to mediate efficacy.
]]></description>
<dc:creator>Barrenäs, F.</dc:creator>
<dc:creator>Hansen, S. G.</dc:creator>
<dc:creator>Law, L.</dc:creator>
<dc:creator>Driscoll, C.</dc:creator>
<dc:creator>Green, R. R.</dc:creator>
<dc:creator>Smith, E.</dc:creator>
<dc:creator>Chang, J.</dc:creator>
<dc:creator>Golez, I.</dc:creator>
<dc:creator>Urion, T.</dc:creator>
<dc:creator>Peng, X.</dc:creator>
<dc:creator>Whitmore, L.</dc:creator>
<dc:creator>Newhouse, D.</dc:creator>
<dc:creator>Hughes, C. M.</dc:creator>
<dc:creator>Morrow, D.</dc:creator>
<dc:creator>Randall, K. T.</dc:creator>
<dc:creator>Selseth, A.</dc:creator>
<dc:creator>Ford, J. C.</dc:creator>
<dc:creator>Gilbride, R. M.</dc:creator>
<dc:creator>Randall, B.</dc:creator>
<dc:creator>Ainslie, E.</dc:creator>
<dc:creator>Oswald, K.</dc:creator>
<dc:creator>Shoemaker, R.</dc:creator>
<dc:creator>Fast, R.</dc:creator>
<dc:creator>Bosche, W. J.</dc:creator>
<dc:creator>Axthelm, M. K.</dc:creator>
<dc:creator>Fukazawa, Y.</dc:creator>
<dc:creator>Pavlakis, G. N.</dc:creator>
<dc:creator>Felber, B. K.</dc:creator>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Sekaly, R.-P.</dc:creator>
<dc:creator>Lifson, J. D.</dc:creator>
<dc:creator>Komorowski, J.</dc:creator>
<dc:creator>Kosmider, E.</dc:creator>
<dc:creator>Shao, J.</dc:creator>
<dc:creator>Song, W.</dc:creator>
<dc:creator>Edlefsen, P. T.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:creator>Gale, M.</dc:creator>
<dc:date>2021-01-11</dc:date>
<dc:identifier>doi:10.1101/2021.01.11.426199</dc:identifier>
<dc:title><![CDATA[Sustained IL-15 response signature predicts RhCMV/SIV vaccine efficacy]]></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/10.1101/2021.01.14.426654v1?rss=1">
<title>
<![CDATA[
MLH1/3 variants causing aneuploidy, pregnancy loss, and premature reproductive aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.14.426654v1?rss=1</link>
<description><![CDATA[
Most spontaneous pregnancy losses are a result of embryonic aneuploidy stemming from mis-segregation of chromosomes during meiosis. Proper disjunction of homologous chromosomes is dependent upon precise control of crossing-over, a process requiring the mismatch repair (MMR) genes MLH1 and MLH3. Both are required for fertility and completion of meiosis in mice. People inheriting variants in these genes are often at high risk for colorectal cancer and Lynch syndrome, yet the potential impacts of variants upon reproduction are unclear. To determine if MLH1/3 variants (namely single nucleotide polymorphisms, or SNPs) in human populations can cause reproductive abnormalities, we used a combination of computational predictions, yeast two-hybrid assays, and assays of MMR and recombination in yeast to select nine MLH1 and MLH3 variants for modeling in mice via genome editing. We identified 7 alleles that caused reproductive defects in mice including subfertility in females, male infertility, reduced sperm counts, and increased spermatocyte apoptosis. Remarkably, these alleles in females caused age-dependent decreases in litter size, and increased resorption of embryos during pregnancy. These outcomes were likely a consequence of reduced meiotic chiasmata, in turn causing an increase in misaligned chromosomes and univalents in meiotic metaphase I (MI). Our data indicate that segregating hypomorphic alleles of meiotic recombination genes in populations can predispose females to increased incidence of pregnancy loss from gamete aneuploidy.
]]></description>
<dc:creator>Fragoza, R.</dc:creator>
<dc:creator>Blengini, C. S.</dc:creator>
<dc:creator>Tran, T. N.</dc:creator>
<dc:creator>Pannafino, G.</dc:creator>
<dc:creator>Al-Sweel, N.</dc:creator>
<dc:creator>Schimenti, K.</dc:creator>
<dc:creator>Schindler, K.</dc:creator>
<dc:creator>Alani, E.</dc:creator>
<dc:creator>Yu, H.</dc:creator>
<dc:creator>Schimenti, J. C.</dc:creator>
<dc:date>2021-01-14</dc:date>
<dc:identifier>doi:10.1101/2021.01.14.426654</dc:identifier>
<dc:title><![CDATA[MLH1/3 variants causing aneuploidy, pregnancy loss, and premature reproductive aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.15.426865v1?rss=1">
<title>
<![CDATA[
DNA-based copy number analysis confirms genomic evolution of PDX models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.15.426865v1?rss=1</link>
<description><![CDATA[
We previously reported the genomic evolution of the copy number (CN) landscapes of patient-derived xenografts (PDXs) during their engraftment and passaging1. Woo et al. argue that the CN profiles of PDXs are highly conserved, and that the main conclusions of our paper are invalid due to our use of expression-based CN profiles2. Here, we reassess genomic evolution of PDXs using the DNA-based CN profiles reported by Woo et al. We find that the degree of genomic evolution in the DNA-based dataset of Woo et al. is similar to that which we had previously reported. While the overall Pearsons correlation of CN profiles between primary tumors (PTs) and their derived PDXs is high (as reported in our original paper as well), a median of ~10% of the genome is differentially altered between PTs and PDXs across cohorts (range, 0% to 73% across all models). In 24% of the matched PT-PDX samples, over a quarter of the genome is differentially affected by CN alterations. Moreover, in matched analyses of PTs and their derived PDXs at multiple passages, later-passage PDXs are significantly less similar to their parental PTs than earlier-passage PDXs, indicative of genomic divergence. We conclude that genomic evolution of PDX models during model generation and propagation should not be dismissed, and that the phenotypic consequences of this evolution ought to be assessed in order to optimize the application of these valuable cancer models.
]]></description>
<dc:creator>Hoge, A.</dc:creator>
<dc:creator>Getz, M.</dc:creator>
<dc:creator>Beroukhim, R.</dc:creator>
<dc:creator>Golub, T.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:creator>Ben-David, U.</dc:creator>
<dc:date>2021-01-17</dc:date>
<dc:identifier>doi:10.1101/2021.01.15.426865</dc:identifier>
<dc:title><![CDATA[DNA-based copy number analysis confirms genomic evolution of PDX models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.23.427919v1?rss=1">
<title>
<![CDATA[
Dynamics and turnover of memory CD8 T cell responses following yellow fever vaccination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.23.427919v1?rss=1</link>
<description><![CDATA[
Understanding how immunological memory lasts a lifetime requires quantifying changes in the number of memory cells as well as how their division and death rates change over time. We address these questions by using a statistically powerful mixed-effects differential equations framework to analyze data from two human studies that follow CD8 T cell responses to the yellow fever vaccine (YFV-17D). Models were first fit to the frequency and division rates of YFV-specific memory CD8 T cells 42 days to 1 year post-vaccination. A different dataset, on the loss of YFV-specific CD8 T cells over three decades, was used to assess out of sample predictions of our models. The commonly used exponential and bi-exponential decline models performed relatively poorly. Models with the cell loss following a power law (exactly or approximately) were most predictive. Notably, using only the first year of data, these models accurately predicted T cell frequencies up to 30 years post-vaccination. Our analyses suggest that division rates of these cells drop and plateau at a low level (0.001 per day, ~double estimates for naive T cells) within one year following vaccination, whereas death rates continue to decline for much longer. Our results show that power laws can be predictive for T cell memory, a finding that may be useful for vaccine evaluation and epidemiological modeling. Moreover, since power laws asymptotically decline more slowly than any exponential decline, our results help explain the longevity of immune memory phenomenologically.

Author summaryImmunological memory, generated in response to infection or vaccination, may provide complete or partial protection from antigenically similar infections for the lifetime. Memory CD8 T cells are important players in protection from secondary viral infections but quantitative understanding of their dynamics in humans is limited. We analyze data from two studies where immunization with the yellow fever virus vaccine (YFV-17D) generates a mild acute infection and long-term memory. We find that: (i) the division rate of YFV-17D-specific CD8 T cells drops and stabilizes at ~ 0.1% per day during the first year following vaccination whereas the death rate declines more gradually, and (ii) the number of these cells declines approximately in accordance with a power law ({propto}time-0.82) for at least several decades following vaccination.
]]></description>
<dc:creator>Zarnitsyna, V. I.</dc:creator>
<dc:creator>Akondy, R. S.</dc:creator>
<dc:creator>Ahmed, H.</dc:creator>
<dc:creator>McGuire, D. J.</dc:creator>
<dc:creator>Zarnitsyn, V. G.</dc:creator>
<dc:creator>Moore, M.</dc:creator>
<dc:creator>Johnson, P. L. F.</dc:creator>
<dc:creator>Ahmed, R.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Hellerstein, M.</dc:creator>
<dc:creator>Antia, R.</dc:creator>
<dc:date>2021-01-23</dc:date>
<dc:identifier>doi:10.1101/2021.01.23.427919</dc:identifier>
<dc:title><![CDATA[Dynamics and turnover of memory CD8 T cell responses following yellow fever vaccination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.25.427999v1?rss=1">
<title>
<![CDATA[
Engineering a niche supporting haematopoietic stem cell development using integrated single cell transcriptomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.25.427999v1?rss=1</link>
<description><![CDATA[
Haematopoietic stem cells (HSCs) develop from haemogenic endothelium (HE) within embryonic arterial vessels such as the aorta of the aorta-gonad-mesonephros region (AGM). To identify the signals responsible for HSC formation, we used single cell RNA-sequencing to simultaneously analyze the transcriptional profiles of AGM-derived cells transitioning from HE to HSC, and AGM-derived endothelial cells which provide signals sufficient to support HSC maturation and self-renewal. Pseudotemporal ordering revealed dynamics of gene expression during the HE to HSC transition, identifying surface receptors specifically expressed on developing HSCs. Transcriptional profiles of niche endothelial cells enabled identification of corresponding ligands, including those signaling to Notch receptors, VLA-4 integrin, and CXCR4, which, when integrated in an engineered platform, were sufficient to support the generation of engrafting HSCs. These studies provide a transcriptional map of the signaling interactions necessary for the development of HSCs and advance the goal of engineering HSC for therapeutic applications.
]]></description>
<dc:creator>Hadland, B.</dc:creator>
<dc:creator>Varnum-Finney, B.</dc:creator>
<dc:creator>Dozono, S.</dc:creator>
<dc:creator>Dignum, T.</dc:creator>
<dc:creator>Nourigat-McKay, C.</dc:creator>
<dc:creator>Jackson, D. L.</dc:creator>
<dc:creator>Itkin, T.</dc:creator>
<dc:creator>Butler, J. M.</dc:creator>
<dc:creator>Rafii, S.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Bernstein, I. D.</dc:creator>
<dc:date>2021-01-26</dc:date>
<dc:identifier>doi:10.1101/2021.01.25.427999</dc:identifier>
<dc:title><![CDATA[Engineering a niche supporting haematopoietic stem cell development using integrated single cell transcriptomics]]></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/10.1101/2021.01.27.428529v1?rss=1">
<title>
<![CDATA[
Genetic and structural basis for recognition of SARS-CoV-2 spike protein by a two-antibody cocktail 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.27.428529v1?rss=1</link>
<description><![CDATA[
The SARS-CoV-2 pandemic has led to an urgent need to understand the molecular basis for immune recognition of SARS-CoV-2 spike (S) glycoprotein antigenic sites. To define the genetic and structural basis for SARS-CoV-2 neutralization, we determined the structures of two human monoclonal antibodies COV2-2196 and COV2-21301, which form the basis of the investigational antibody cocktail AZD7442, in complex with the receptor binding domain (RBD) of SARS-CoV-2. COV2-2196 forms an "aromatic cage" at the heavy/light chain interface using germline-encoded residues in complementarity determining regions (CDRs) 2 and 3 of the heavy chain and CDRs 1 and 3 of the light chain. These structural features explain why highly similar antibodies (public clonotypes) have been isolated from multiple individuals1-4. The structure of COV2-2130 reveals that an unusually long LCDR1 and HCDR3 make interactions with the opposite face of the RBD from that of COV2-2196. Using deep mutational scanning and neutralization escape selection experiments, we comprehensively mapped the critical residues of both antibodies and identified positions of concern for possible viral escape. Nonetheless, both COV2-2196 and COV2-2130 showed strong neutralizing activity against SARS-CoV-2 strain with recent variations of concern including E484K, N501Y, and D614G substitutions. These studies reveal germline-encoded antibody features enabling recognition of the RBD and demonstrate the activity of a cocktail like AZD7442 in preventing escape from emerging variant viruses.
]]></description>
<dc:creator>Dong, J.</dc:creator>
<dc:creator>Zost, S.</dc:creator>
<dc:creator>Greaney, A.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Chen, E. C.</dc:creator>
<dc:creator>Chen, R.</dc:creator>
<dc:creator>Case, B.</dc:creator>
<dc:creator>Sutton, R.</dc:creator>
<dc:creator>Gilchuk, P.</dc:creator>
<dc:creator>Rodriguez, J.</dc:creator>
<dc:creator>Armstrong, E.</dc:creator>
<dc:creator>Gainza, C.</dc:creator>
<dc:creator>Nargi, R.</dc:creator>
<dc:creator>Binshtein, E.</dc:creator>
<dc:creator>Xie, X.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Shi, P.-Y.</dc:creator>
<dc:creator>Logue, J.</dc:creator>
<dc:creator>Weston, S.</dc:creator>
<dc:creator>McGrath, M.</dc:creator>
<dc:creator>Frieman, M.</dc:creator>
<dc:creator>Brady, T.</dc:creator>
<dc:creator>Tuffy, K.</dc:creator>
<dc:creator>Bright, H.</dc:creator>
<dc:creator>Loo, Y.-M.</dc:creator>
<dc:creator>McvTamney, P.</dc:creator>
<dc:creator>Esser, M.</dc:creator>
<dc:creator>Carnahan, R.</dc:creator>
<dc:creator>Diamond, M.</dc:creator>
<dc:creator>Bloom, J.</dc:creator>
<dc:creator>Crowe, J. E.</dc:creator>
<dc:date>2021-01-28</dc:date>
<dc:identifier>doi:10.1101/2021.01.27.428529</dc:identifier>
<dc:title><![CDATA[Genetic and structural basis for recognition of SARS-CoV-2 spike protein by a two-antibody cocktail]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.28.428695v1?rss=1">
<title>
<![CDATA[
RSC primes the quiescent genome for hypertranscription upon cell cycle re-entry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.28.428695v1?rss=1</link>
<description><![CDATA[
Quiescence is a reversible G0 state essential for differentiation, regeneration, stem cell renewal, and immune cell activation. Necessary for long-term survival, quiescent chromatin is compact, hypoacetylated, and transcriptionally inactive. How transcription activates upon cell-cycle re-entry is undefined. Here we report robust, widespread transcription within the first minutes of quiescence exit. During quiescence, the chromatin-remodeling enzyme RSC was already bound to the genes induced upon quiescence exit. RSC depletion caused severe quiescence exit defects: a global decrease in RNA polymerase II (Pol II) loading, Pol II accumulation at transcription start sites, initiation from ectopic upstream loci, and aberrant antisense transcription. These phenomena were due to a combination of highly robust Pol II transcription and severe chromatin defects in the promoter regions and gene bodies. Together, these results uncovered multiple mechanisms by which RSC facilitates initiation and maintenance of large-scale, rapid gene expression despite a globally repressive chromatin state.
]]></description>
<dc:creator>Cucinotta, C. E.</dc:creator>
<dc:creator>Dell, R. H.</dc:creator>
<dc:creator>Braceros, K. C. A.</dc:creator>
<dc:creator>Tsukiyama, T.</dc:creator>
<dc:date>2021-01-28</dc:date>
<dc:identifier>doi:10.1101/2021.01.28.428695</dc:identifier>
<dc:title><![CDATA[RSC primes the quiescent genome for hypertranscription upon cell cycle re-entry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.29.428899v1?rss=1">
<title>
<![CDATA[
A Cas-BCAR3 co-regulatory circuit controls lamellipodia dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.29.428899v1?rss=1</link>
<description><![CDATA[
Integrin adhesion complexes regulate cytoskeletal dynamics during cell migration. Adhesion activates phosphorylation of integrin-associated signaling proteins, including Cas (p130Cas, BCAR1), by Src-family kinases. Cas regulates leading-edge protrusion and migration in cooperation with its binding partner, BCAR3. However, it has been unclear how Cas and BCAR3 cooperate. Here, using normal epithelial cells, we find that BCAR3 localization to integrin adhesions requires Cas. In return, Cas phosphorylation, as well as lamellipodia dynamics and cell migration, requires BCAR3. These functions require the BCAR3 SH2 domain and a specific phosphorylation site, Tyr 117, that is also required for BCAR3 downregulation by the ubiquitin-proteasome system. These findings place BCAR3 in a co-regulatory positive-feedback circuit with Cas, with BCAR3 requiring Cas for localization and Cas requiring BCAR3 for activation and downstream signaling. The use of a single phosphorylation site in BCAR3 for activation and degradation ensures reliable negative feedback by the ubiquitin-proteasome system.
]]></description>
<dc:creator>Steenkiste, E. M.</dc:creator>
<dc:creator>Berndt, J. D.</dc:creator>
<dc:creator>Pilling, C.</dc:creator>
<dc:creator>Simpkins, C.</dc:creator>
<dc:creator>Cooper, J. A.</dc:creator>
<dc:date>2021-01-29</dc:date>
<dc:identifier>doi:10.1101/2021.01.29.428899</dc:identifier>
<dc:title><![CDATA[A Cas-BCAR3 co-regulatory circuit controls lamellipodia dynamics]]></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/10.1101/2021.01.29.428913v1?rss=1">
<title>
<![CDATA[
Atg8 licenses adipokine nuclear exit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.29.428913v1?rss=1</link>
<description><![CDATA[
Adipokines released from the adipocytes function as a systemic adipometer; they impinge on neural circuits to signal nutrient status. On starvation, adipokines must be retained to signal energy deficit; else, it significantly reduces starvation survival. But how fat cells retain adipokines is unclear. Here, we demonstrate that Atg8, a cell-intrinsic autophagy factor, regulates the starvation-induced acute retention of the Leptin Drosophila ortholog Upd2. We show that on starvation, as a direct consequence of Atg8s lipidation, Upd2 accumulates in the nucleus. We illustrate that Upd2s nuclear retention is critical to fat mobilization and increased starvation resilience. Furthermore, nuclear Upd2 promotes the expression of a secreted innate immune gene signature. This hints at an unanticipated connection between adipokine nuclear retention and increased innate immunity. In conclusion, we propose that, during starvation, Atg8s role is not just limited to autophagy but is critical for withholding adipokines in the nucleus to promote starvation resilience.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=178 HEIGHT=200 SRC="FIGDIR/small/428913v5_ufig1.gif" ALT="Figure 1">
View larger version (33K):
org.highwire.dtl.DTLVardef@1826425org.highwire.dtl.DTLVardef@17cca52org.highwire.dtl.DTLVardef@1c70ac7org.highwire.dtl.DTLVardef@94a024_HPS_FORMAT_FIGEXP  M_FIG C_FIG O_LIIn fed state Upd2 requires Atg8 for nuclear exit and cytosolic localization.
C_LIO_LIAtg8s lipidation on starvation results in Upd2s nuclear accumulation.
C_LIO_LIUpd2 nuclear retention on starvation increases fat mobilization and post-starvation hunger.
C_LIO_LIOn starvation Upd2 nuclear retention increases expression of a secreted innate immune signature.
C_LI
]]></description>
<dc:creator>Poling, M. E.</dc:creator>
<dc:creator>Sullivan, C. E.</dc:creator>
<dc:creator>Brent, A. E.</dc:creator>
<dc:creator>Hafer, T. L.</dc:creator>
<dc:creator>Rajan, A.</dc:creator>
<dc:date>2021-01-31</dc:date>
<dc:identifier>doi:10.1101/2021.01.29.428913</dc:identifier>
<dc:title><![CDATA[Atg8 licenses adipokine nuclear exit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.09.430515v1?rss=1">
<title>
<![CDATA[
Signaling amplitude molds the Ras mutation tropism of urethane 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.09.430515v1?rss=1</link>
<description><![CDATA[
RAS genes are commonly mutated in cancers yet despite many possible mutations, cancers have a  tropism towards a specific subset. As driver mutations, these patterns ostensibly originate from normal cells. High oncogenic RAS activity causes oncogenic stress and different oncogenic mutations can impart different levels of activity. Here we show that changing rare codons to common in the murine Kras gene to increase translation shifts tumors induced by the carcinogen urethane from arising from canonical Q61 to biochemically less active G12 Kras driver mutations, despite the carcinogen still being biased towards generating Q61 mutations. Loss of p53 to blunt oncogenic stress partially reversed this effect, restoring Q61 mutations. Finally, transcriptional analysis revealed similar signaling amongst tumors driven by different mutations and Kras alleles. These finding suggest that the RAS mutation tropism of urethane is largely product of selection in normal cells for mutations promoting proliferation without causing oncogenic stress.

Impact statementThe bias towards specific Kras driver mutations during urethane carcinogenesis appears to arise predominantly from the selection of a narrow window of oncogenic signaling in normal cells.
]]></description>
<dc:creator>Counter, C.</dc:creator>
<dc:creator>Li, s.</dc:creator>
<dc:date>2021-02-10</dc:date>
<dc:identifier>doi:10.1101/2021.02.09.430515</dc:identifier>
<dc:title><![CDATA[Signaling amplitude molds the Ras mutation tropism of urethane]]></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/10.1101/2021.02.12.430999v1?rss=1">
<title>
<![CDATA[
An improved ChEC-seq method accurately maps the genome-wide binding of transcription coactivators and sequence-specific transcription factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.12.430999v1?rss=1</link>
<description><![CDATA[
Mittal and colleagues have raised questions about mapping transcription factor locations on DNA using the MNase-based ChEC-seq method (Mittal et al., 2021). Partly due to this concern, we modified the experimental conditions of the MNase cleavage step and subsequent computational analyses, resulting in more stringent conditions for mapping protein-DNA interactions (Donczew et al., 2020). The revised method (dx.doi.org/10.17504/protocols.io.bizgkf3w) answers questions raised by Mittal et al. and, without changing earlier conclusions, identified widespread promoter binding of the transcription coactivators TFIID and SAGA at active genes. The revised method is also suitable for accurately mapping the genome-wide locations of DNA sequence-specific transcription factors.
]]></description>
<dc:creator>Donczew, R.</dc:creator>
<dc:creator>Lalou, A.</dc:creator>
<dc:creator>Devys, D.</dc:creator>
<dc:creator>Tora, L.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:date>2021-02-12</dc:date>
<dc:identifier>doi:10.1101/2021.02.12.430999</dc:identifier>
<dc:title><![CDATA[An improved ChEC-seq method accurately maps the genome-wide binding of transcription coactivators and sequence-specific transcription factors]]></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/10.1101/2021.02.16.431009v1?rss=1">
<title>
<![CDATA[
PhylEx: Accurate reconstruction of clonal structure via integrated analysis of bulk DNA-seq and single cell RNA-seq data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.16.431009v1?rss=1</link>
<description><![CDATA[
We propose PhylEx: a clonal-tree reconstruction method that integrates bulk genomics and single-cell transcriptomics data. In addition to the clonal-tree, PhylEx also assigns single-cells to clones, which effectively produce clonal expression profiles, and generates clonal genotypes. By analyzing scRNA-seq integrated with bulk DNA-seq, PhylEx can take advantage of co-occurrences of the mutations found in the cells. In the probabilistic model underlying PhylEx, the raw read counts from scRNA-seq follow a mixture of Beta-Binomial distributions, which accounts for the sparse nature of single-cell gene expression data; the mixture lessens the penalty caused by mutations not observed due to mono-allelic expression. We rigorously evaluated PhylEx on simulated datasets as well as a biological dataset consisting of a previously well-characterized high-grade serous ovarian cancer (HGSOC) cell line. PhylEx outperformed the state-of-the-art methods by a wide margin both when comparing capacity for clonal-tree reconstruction and capacity for correctly clustering mutations. By analyzing HGSOC and HER2+ breast cancer data, we also show that PhylEx clears the way for phylo-phenotypic analysis of cancer, i.e., that the clonal expression profiles, induced by the cell-to-clone assignments, can be exploited in a manner beyond what is possible with only expression-based clustering.
]]></description>
<dc:creator>Jun, S.-H.</dc:creator>
<dc:creator>Toosi, H.</dc:creator>
<dc:creator>Mold, J.</dc:creator>
<dc:creator>Engblom, C.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>O'Flanagan, C.</dc:creator>
<dc:creator>Hagemann-Jensen, M.</dc:creator>
<dc:creator>Sandberg, R.</dc:creator>
<dc:creator>Aparicio, S.</dc:creator>
<dc:creator>Hartman, J.</dc:creator>
<dc:creator>Roth, A.</dc:creator>
<dc:creator>Lagergren, J.</dc:creator>
<dc:date>2021-02-17</dc:date>
<dc:identifier>doi:10.1101/2021.02.16.431009</dc:identifier>
<dc:title><![CDATA[PhylEx: Accurate reconstruction of clonal structure via integrated analysis of bulk DNA-seq and single cell RNA-seq data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.17.431683v1?rss=1">
<title>
<![CDATA[
Complete map of SARS-CoV-2 RBD mutations that escape the monoclonal antibody LY-CoV555 and its cocktail with LY-CoV016 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.17.431683v1?rss=1</link>
<description><![CDATA[
Monoclonal antibodies and antibody cocktails are a promising therapeutic and prophylaxis for COVID-19. However, ongoing evolution of SARS-CoV-2 can render monoclonal antibodies ineffective. Here we completely map all mutations to the SARS-CoV-2 spike receptor binding domain (RBD) that escape binding by a leading monoclonal antibody, LY-CoV555, and its cocktail combination with LY-CoV016. Individual mutations that escape binding by each antibody are combined in the circulating B.1.351 and P.1 SARS-CoV-2 lineages (E484K escapes LY-CoV555, K417N/T escape LY-CoV016). Additionally, the L452R mutation in the B.1.429 lineage escapes LY-CoV555. Furthermore, we identify single amino acid changes that escape the combined LY-CoV555+LY-CoV016 cocktail. We suggest that future efforts should diversify the epitopes targeted by antibodies and antibody cocktails to make them more resilient to antigenic evolution of SARS-CoV-2.
]]></description>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-02-22</dc:date>
<dc:identifier>doi:10.1101/2021.02.17.431683</dc:identifier>
<dc:title><![CDATA[Complete map of SARS-CoV-2 RBD mutations that escape the monoclonal antibody LY-CoV555 and its cocktail with LY-CoV016]]></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/10.1101/2021.02.17.431715v1?rss=1">
<title>
<![CDATA[
Dopamine receptor DOP-1 engages a sleep pathway to modulate swimming in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.17.431715v1?rss=1</link>
<description><![CDATA[
Animals require robust yet flexible programs to support locomotion. While it is clear that a variety of processes must be engaged to ensure rhythmic actions, the exact mechanisms remain largely unknown. Here we report a novel pathway that connects the D1-like dopamine receptor DOP-1 with a sleep mechanism to modulate swimming in C. elegans. We show that DOP-1 plays a negative role in sustaining swimming behavior. By contrast, a pathway through the D2-like dopamine receptor DOP-3 negatively regulates the initiation of swimming, but its impact fades quickly over a few minutes. We find that DOP-1 and the GPCR kinase GRK-2 function in the sleep interneuron RIS, where DOP-1 modulates the secretion of a sleep neuropeptide FLP-11. Our genetic studies further show that DOP-1 and FLP-11 act in the same pathway to modulate swimming. Together, these results delineate a functional connection between a dopamine receptor and a sleep program to regulate swimming in C. elegans. The temporal transition between DOP-3 and DOP-1 pathways highlights the dynamic nature of neuromodulation for rhythmic movements that persist over time.

HIGHLIGHTSO_LIThe D1-like dopamine receptor DOP-1 regulates swimming at 10 minutes
C_LIO_LIAn integrated function of DOP-1 and DOP-3 is required for the continuity of swimming
C_LIO_LIDOP-1 and GRK-2 act in the sleep neuron RIS
C_LIO_LIFLP-11, a neuropeptide that promotes sleep, negatively regulates swimming
C_LI

IN BRIEFXu et al. investigated genetic programs that modulate swimming behavior in the nematode C. elegans. They identified a functional link that couples a D1-like dopamine receptor to a sleep program that modulates the sustained phase rather than the initial phase of swimming.
]]></description>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Topalidou, I.</dc:creator>
<dc:creator>Hassinan, C.</dc:creator>
<dc:creator>Ailion, M.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Bai, J.</dc:creator>
<dc:date>2021-02-18</dc:date>
<dc:identifier>doi:10.1101/2021.02.17.431715</dc:identifier>
<dc:title><![CDATA[Dopamine receptor DOP-1 engages a sleep pathway to modulate swimming in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.18.431862v1?rss=1">
<title>
<![CDATA[
Pregnancy and weaning regulate human maternal liver size and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.18.431862v1?rss=1</link>
<description><![CDATA[
BACKGROUNDDuring pregnancy, the rodent liver undergoes hepatocyte proliferation and increases in size, followed by weaning-induced involution via hepatocyte cell death and stromal remodeling, creating a pro-metastatic niche. These data suggest a mechanism for increased liver metastasis in postpartum breast cancer patients.

OBJECTIVESInvestigate if the human liver changes in size and function during pregnancy and weaning.

METHODSAbdominal imaging was obtained in healthy women at early and late pregnancy, and post-wean. During pregnancy time points, endogenous glucose production was measured and fasting blood taken to measure bile acids.

RESULTSIndependent of weight gain, most womens livers increased in size with pregnancy, returning to baseline post-wean. Putative roles for bile acids in liver growth were observed in pregnant women and rodents.

CONCLUSIONSThe human liver is regulated by reproductive state with growth during pregnancy and volume loss post-wean. These findings may have broad implications for sex-specific liver diseases and cancer.
]]></description>
<dc:creator>Bartlett, A. Q.</dc:creator>
<dc:creator>Francisco, M.</dc:creator>
<dc:creator>Goddard, E.</dc:creator>
<dc:creator>DeBarber, A.</dc:creator>
<dc:creator>Leo, M. C.</dc:creator>
<dc:creator>Baetscher, E.</dc:creator>
<dc:creator>Rooney, W.</dc:creator>
<dc:creator>Naugler, W.</dc:creator>
<dc:creator>Catalano, P.</dc:creator>
<dc:creator>Guimaraes, A.</dc:creator>
<dc:creator>Purnell, J. Q.</dc:creator>
<dc:creator>Vesco, K. K.</dc:creator>
<dc:creator>Schedin, P. J.</dc:creator>
<dc:date>2021-02-18</dc:date>
<dc:identifier>doi:10.1101/2021.02.18.431862</dc:identifier>
<dc:title><![CDATA[Pregnancy and weaning regulate human maternal liver size and function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.23.432444v1?rss=1">
<title>
<![CDATA[
A large-scale transcriptome-wide association study (TWAS) of ten blood cell phenotypes reveals complexities of TWAS fine-mapping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.23.432444v1?rss=1</link>
<description><![CDATA[
Hematological measures are important intermediate clinical phenotypes for many acute and chronic diseases. Hematological measures are highly heritable, and although genome-wide association studies (GWAS) have identified thousands of loci containing trait-associated variants, the causal genes underlying these associations are often uncertain. To better understand the underlying genetic regulatory mechanisms, we performed a transcriptome-wide association study (TWAS) using PrediXcan to systematically investigate the association between genetically-predicted gene expression and hematological measures in 54,542 individuals of European ancestry from the Genetic Epidemiology Research on Adult Health and Aging (GERA) cohort. We found 239 significant gene-trait associations with hematological measures. Among this set of 239 associations, we replicated 71 at p < 0.05 with same direction of effect for the blood cell trait in a meta-analysis of TWAS results consisting of up to 35,900 European ancestry individuals from the Womens Health Initiative (WHI), the Atherosclerosis Risk in Communities Study (ARIC), and BioMe Biobank. We further attempted to refine this list of candidate genes by performing conditional analyses, adjusting for individual variants previously associated with these hematological measures, and performed further fine-mapping of TWAS loci. To assist with the interpretation of TWAS findings, we designed an R Shiny application to interactively visualize TWAS results, one genomic locus at a time, by integrating our TWAS results with additional genetic data sources (GWAS, TWAS from other gene expression reference panels, conditional analyses, known GWAS variants, etc.). Our results and R Shiny application highlight frequently overlooked challenges with TWAS and illustrate the complexity of TWAS fine-mapping efforts.

Author SummaryTranscriptome-wide association studies (TWAS) have shown great promise in furthering our understanding of the genetic regulatory mechanisms underlying complex trait variation. However, interpreting TWAS results can be incredibly complex, especially in large-scale analyses where hundreds of signals appear throughout the genome, with multiple genes often identified in a single chromosomal region. Our research demonstrates this complexity through real data examples from our analysis of hematological traits, and we provide a useful web application to visualize TWAS results in a broadly approachable format. Together, our results and web application illustrate the importance of interpreting TWAS studies in context and highlight the need to carefully examine results in a region-wide context to draw reasonable conclusions and formulate mechanistic hypotheses.
]]></description>
<dc:creator>Tapia, A. L.</dc:creator>
<dc:creator>Rowland, B. T.</dc:creator>
<dc:creator>Rosen, J. D.</dc:creator>
<dc:creator>Preuss, M.</dc:creator>
<dc:creator>Young, K.</dc:creator>
<dc:creator>Graff, M.</dc:creator>
<dc:creator>Choquet, H.</dc:creator>
<dc:creator>Couper, D. J.</dc:creator>
<dc:creator>Buyske, S.</dc:creator>
<dc:creator>Bien, S. A.</dc:creator>
<dc:creator>Jorgenson, E.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Loos, R. J. F.</dc:creator>
<dc:creator>Morrison, A. C.</dc:creator>
<dc:creator>North, K. E.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Raffield, L. M.</dc:creator>
<dc:date>2021-02-23</dc:date>
<dc:identifier>doi:10.1101/2021.02.23.432444</dc:identifier>
<dc:title><![CDATA[A large-scale transcriptome-wide association study (TWAS) of ten blood cell phenotypes reveals complexities of TWAS fine-mapping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.24.432809v1?rss=1">
<title>
<![CDATA[
Odor Blocking Of Stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.24.432809v1?rss=1</link>
<description><![CDATA[
Scents have been employed for millennia to allay fear and stress, but whether they do so is poorly understood. In response to fear and stress, hypothalamic corticotropin releasing hormone neurons (CRHNs) induce increases in blood stress hormones. Here, we find that certain structurally and perceptually dissimilar odorants can block mouse stress hormone responses to three potent stressors: physical restraint, predator odor, and male-male social confrontation. Both odorants activate GABAergic inhibitory neurons presynaptic to CRHNs in the hypothalamic ventromedial nucleus (VMH). Stimulation of those neurons inhibits restraint-induced activation of CRHNs and stress hormone increase, mimicking a blocking odorant. Conversely, their silencing prevents odorant blocking of both responses. Notably, we also observed odor blocking of stressor activation in neurons presynaptic to CRHNs in the bed nucleus of the stria terminalis (BNST). Together, these findings indicate that selected odorants can indeed block stress responses, and that odor blocking can occur via two routes: a direct route in which blocking odor signals directly inhibit CRHNs and an indirect route in which they inhibit stressor activation of neurons presynaptic to CRHNs and prevent them from transmitting stress signals to CRHNs.
]]></description>
<dc:creator>Lee, E. J.</dc:creator>
<dc:creator>Saraiva, L. R.</dc:creator>
<dc:creator>Hanchate, N. K.</dc:creator>
<dc:creator>Ye, X.</dc:creator>
<dc:creator>Asher, G.</dc:creator>
<dc:creator>Ho, J.</dc:creator>
<dc:creator>Buck, L. B.</dc:creator>
<dc:date>2021-02-25</dc:date>
<dc:identifier>doi:10.1101/2021.02.24.432809</dc:identifier>
<dc:title><![CDATA[Odor Blocking Of Stress]]></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/10.1101/2021.02.25.432905v1?rss=1">
<title>
<![CDATA[
A public broadly neutralizing antibody class targets a membrane-proximal anchor epitope of influenza virus hemagglutinin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.25.432905v1?rss=1</link>
<description><![CDATA[
Broadly neutralizing antibodies against influenza virus hemagglutinin (HA) have the potential to provide universal protection against influenza virus infections. Here, we report a distinct class of broadly neutralizing antibodies targeting an epitope toward the bottom of the HA stalk domain where HA is "anchored" to the viral membrane. Antibodies targeting this membrane-proximal anchor epitope utilized a highly restricted repertoire, which encode for two conserved motifs responsible for HA binding. Anchor targeting B cells were common in the human memory B cell repertoire across subjects, indicating pre-existing immunity against this epitope. Antibodies against the anchor epitope at both the serological and monoclonal antibody levels were potently induced in humans by a chimeric HA vaccine, a potential universal influenza virus vaccine. Altogether, this study reveals an underappreciated class of broadly neutralizing antibodies against H1-expressing viruses that can be robustly recalled by a candidate universal influenza virus vaccine.
]]></description>
<dc:creator>Guthmiller, J. J.</dc:creator>
<dc:creator>Han, J.</dc:creator>
<dc:creator>Utset, H. A.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Lan, L. Y.-L.</dc:creator>
<dc:creator>Henry, C.</dc:creator>
<dc:creator>Stamper, C. T.</dc:creator>
<dc:creator>Stovicek, O.</dc:creator>
<dc:creator>Gentles, L.</dc:creator>
<dc:creator>Dugan, H. L.</dc:creator>
<dc:creator>Zheng, N.-Y.</dc:creator>
<dc:creator>Richey, S. T.</dc:creator>
<dc:creator>Tepora, M. E.</dc:creator>
<dc:creator>Bitar, D. J.</dc:creator>
<dc:creator>Changrob, S.</dc:creator>
<dc:creator>Strohmeier, S.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Garcia-Sastre, A.</dc:creator>
<dc:creator>Nachbagauer, R.</dc:creator>
<dc:creator>Palese, P.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Krammer, F.</dc:creator>
<dc:creator>Coughlan, L.</dc:creator>
<dc:creator>Ward, A. B.</dc:creator>
<dc:creator>Wilson, P. C.</dc:creator>
<dc:date>2021-02-27</dc:date>
<dc:identifier>doi:10.1101/2021.02.25.432905</dc:identifier>
<dc:title><![CDATA[A public broadly neutralizing antibody class targets a membrane-proximal anchor epitope of influenza virus hemagglutinin]]></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/10.1101/2021.03.02.432825v1?rss=1">
<title>
<![CDATA[
Factors influencing maternal microchimerism throughout infancy and its impact on infant T cell immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.02.432825v1?rss=1</link>
<description><![CDATA[
Determinants of the acquisition and maintenance of maternal microchimerism (MMc) during infancy and the impact of MMc on infant immune responses are unknown. We examined factors which influence MMc detection and level across infancy and the effect of MMc on T cell responses to BCG vaccination in a cohort of HIV exposed, uninfected and HIV unexposed infants in South Africa. MMc was measured in whole blood from 58 infants using a panel of quantitative PCR assays at day one and 7, 15, and 36 weeks of life. Infants received BCG at birth, and selected whole blood samples from infancy were stimulated in vitro with BCG and assessed for polyfunctional CD4+ T cell responses. MMc was present in most infants across infancy with levels ranging from 0-1,193/100,000 genomic equivalents and was positively impacted by absence of maternal HIV, maternal-infant HLA compatibility, infant female sex, and exclusive breastfeeding. Initiation of maternal antiretroviral therapy prior to pregnancy partially restored MMc level in HIV exposed, uninfected infants. Birth MMc was associated with an improved polyfunctional CD4+ T cell response to BCG. These data emphasize that both maternal and infant factors influence the level of MMc, which may subsequently impact infant T cell responses.

O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY
]]></description>
<dc:creator>Balle, C.</dc:creator>
<dc:creator>Kiravu, A.</dc:creator>
<dc:creator>Hoffmann, A.</dc:creator>
<dc:creator>Happel, A.-U.</dc:creator>
<dc:creator>Kanaan, S. B.</dc:creator>
<dc:creator>Nelson, J. L.</dc:creator>
<dc:creator>Gray, C.</dc:creator>
<dc:creator>Jaspan, H. B.</dc:creator>
<dc:creator>Harrington, W. E.</dc:creator>
<dc:date>2021-03-02</dc:date>
<dc:identifier>doi:10.1101/2021.03.02.432825</dc:identifier>
<dc:title><![CDATA[Factors influencing maternal microchimerism throughout infancy and its impact on infant T cell immunity]]></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/10.1101/2021.03.08.434490v1?rss=1">
<title>
<![CDATA[
Differential condensation of sister chromatids coordinates with Cdc6 to ensure distinct cell cycle progression in Drosophila male germline stem cell lineage 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.08.434490v1?rss=1</link>
<description><![CDATA[
Stem cells undergo asymmetric division to produce both a self-renewing stem cell and a differentiating daughter cell. During Drosophila male germline stem cell (GSC) asymmetric division, preexisting old histones H3 and H4 are enriched in the self-renewed stem daughter cell, whereas the newly synthesized H3 and H4 are enriched in the differentiating daughter cell. However, the biological consequences in the two daughter cells resulting from asymmetric histone inheritance remained to be elucidated. In this work, we track both old and new histones throughout GSC cell cycle using high spatial and temporal resolution microscopy. We find several unique features differentiating old versus new histone-enriched sister chromatids, including nucleosome density, chromosomal condensation, and H3 Ser10 phosphorylation. These distinct chromosomal features lead to their differential association with Cdc6, an essential component of the pre-replication complex, which subsequently contributes to asynchronous initiation of DNA replication in the two resulting daughter cells. Disruption of asymmetric histone inheritance abolishes both differential Cdc6 association and asynchronous S-phase entry, demonstrating that asymmetric histone acts upstream of these critical events during cell cycle progression. Furthermore, GSC defects are detected under these conditions, indicating a connection between histone inheritance, cell cycle progression and cell fate decision. Together, these studies reveal that cell cycle remodeling as a crucial biological  readout of asymmetric histone inheritance, which precedes and could lead to other well-known readouts such as differential gene expression. This work also enhances our understanding of asymmetric histone inheritance and epigenetic regulation in other stem cells or asymmetrically dividing cells in multicellular organisms.
]]></description>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Ranjan, R.</dc:creator>
<dc:creator>Snedeker, J.</dc:creator>
<dc:creator>Wooten, M.</dc:creator>
<dc:creator>Chu, C.</dc:creator>
<dc:creator>Bracer, S.</dc:creator>
<dc:creator>Mouton, T.</dc:creator>
<dc:date>2021-03-09</dc:date>
<dc:identifier>doi:10.1101/2021.03.08.434490</dc:identifier>
<dc:title><![CDATA[Differential condensation of sister chromatids coordinates with Cdc6 to ensure distinct cell cycle progression in Drosophila male germline stem cell lineage]]></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/10.1101/2021.03.09.434681v1?rss=1">
<title>
<![CDATA[
RNA-silencing induces target gene relocalization toward a specialized nuage domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.09.434681v1?rss=1</link>
<description><![CDATA[
The authors wish to withdraw this manuscript and apologize for errors in the initial submission. All the original experiments were performed by YY. Unfortunately, JP and members of the Mello lab have not been able to replicate some aspects of the study. JP has failed to independently reproduce the specific results showing RNAi-triggered relocalization of target RNA, and P granule specific accumulation of (the P granule component GLH-1) as reported. The conditions/strains analyzed by JP were as follows: (1) oma-1 FISH on WT worms [control, 6 hr and 12 hr oma-1(RNAi)]. (2) oma-1 FISH on OMA-1:GFP worms [control, 6 hr oma-1(RNAi), or 6 hr gfp(RNAi)]. (3) oma-1 FISH on WT worms [control, 4hr, 6hr, 8 hr, and 10 hr oma-1(RNAi)]. 10-23 gonads were analyzed per experiment. Fixation conditions were essentially as described, with the only known difference being that gonads were not exposed to detergent prior to fixation. Using YYs reagents and protocol the Mello lab has not observed an obvious relocalization of target RNA to P granules (marked by GFP::GLH-1) after 6 hrs oma-1(RNAi); n=92 gonads. CM, JP and DG consider that the published images accurately represent the image stacks provided by YY as representative, raw data, but JP and CM note configurations of FISH signals in germ nuclei and gonad anatomy that they consider unusual. CM, JP and DG have not detected any evidence of image manipulation.

YY states that none of the raw image data were manipulated beyond standard adjustments for brightness and contrast prior to processing images for publication as described. However, YY reports that the images were not representative of the majority of sample gonads, and instead were pre-selected under low magnification for rare examples with asymmetrical, expanded P granules.

Efforts to identify conditions that explain the rare gonads imaged by YY continue in the Mello lab, as do efforts to reproduce independently each of the other reported results; we plan to provide an update in the near future.
]]></description>
<dc:creator>Mello, C.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Grunwald, D.</dc:creator>
<dc:creator>Priess, J.</dc:creator>
<dc:date>2021-03-11</dc:date>
<dc:identifier>doi:10.1101/2021.03.09.434681</dc:identifier>
<dc:title><![CDATA[RNA-silencing induces target gene relocalization toward a specialized nuage domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.10.434844v1?rss=1">
<title>
<![CDATA[
Cell-type-specific chromatin occupancy by the pioneer factor Zelda drives key developmental transitions in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.10.434844v1?rss=1</link>
<description><![CDATA[
During Drosophila embryogenesis, the essential pioneer factor Zelda defines hundreds of cis-regulatory regions and in doing so reprograms the zygotic transcriptome. While Zelda is essential later in development, it is unclear how the ability of Zelda to define cis-regulatory regions is shaped by cell-type-specific chromatin architecture. Asymmetric division of neural stem cells (neuroblasts) in the fly brain provide an excellent paradigm for investigating the cell-type-specific functions of this pioneer factor. We show that Zelda synergistically functions with Notch to maintain neuroblasts in an undifferentiated state. Zelda misexpression reprograms progenitor cells to neuroblasts, but this capacity is limited by transcriptional repressors critical for progenitor commitment. Zelda genomic occupancy in neuroblasts is reorganized as compared to the embryo, and this reorganization is driven by differences in chromatin accessibility and cofactor availability. We propose that Zelda regulates essential transitions in the neuroblasts and embryo through a shared gene-regulatory network by defining cell-type-specific enhancers.
]]></description>
<dc:creator>Larson, E. D.</dc:creator>
<dc:creator>Komori, H.</dc:creator>
<dc:creator>Gibson, T. J.</dc:creator>
<dc:creator>Ostgaard, C. M.</dc:creator>
<dc:creator>Hamm, D. C.</dc:creator>
<dc:creator>Schnell, J. M.</dc:creator>
<dc:creator>Lee, C.-Y.</dc:creator>
<dc:creator>Harrison, M. M.</dc:creator>
<dc:date>2021-03-10</dc:date>
<dc:identifier>doi:10.1101/2021.03.10.434844</dc:identifier>
<dc:title><![CDATA[Cell-type-specific chromatin occupancy by the pioneer factor Zelda drives key developmental transitions in Drosophila]]></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/10.1101/2021.03.15.435484v1?rss=1">
<title>
<![CDATA[
Functional development of a V3-specific broadly neutralizing antibody isolated from a case of HIV superinfection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.15.435484v1?rss=1</link>
<description><![CDATA[
Stimulating broadly neutralizing antibodies (bnAbs) directly from germline remains a barrier for HIV vaccines. HIV superinfection elicits bnAbs more frequently than single infection, providing clues of how to elicit such responses. We used longitudinal antibody sequencing and structural studies to characterize bnAb development from a superinfection case. Mature bnAb QA013.2 bound both initial transmitted and superinfecting virus, but its inferred naive bound only the superinfecting strain and was not neutralizing. QA013.2 requires residues spanning FWRH1-CDRH1 to attain breadth, which is uncommon for V3-specific bnAbs. A 4.15 [A] cryo-EM structure of QA013.2 bound to heterologous native-like trimer showed recognition of V3 signatures (N301, N332, and GDIR). Antigenic profiling revealed that viral escape was achieved not only by changes in the structurally-defined epitope, but also by mutations in V1. These results highlight shared and distinct properties of QA013.2 relative to other V3-specific bnAbs in the setting of sequential, diverse antigenic variants.
]]></description>
<dc:creator>Shipley, M. M.</dc:creator>
<dc:creator>Mangala Prasad, V.</dc:creator>
<dc:creator>Doepker, L. E.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Ralph, D. K.</dc:creator>
<dc:creator>Harkins, E.</dc:creator>
<dc:creator>Dhar, A.</dc:creator>
<dc:creator>Arenz, D.</dc:creator>
<dc:creator>Chohan, V.</dc:creator>
<dc:creator>Weight, H.</dc:creator>
<dc:creator>Mandaliya, K.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Matsen, F.</dc:creator>
<dc:creator>Lee, K. K.</dc:creator>
<dc:creator>Overbaugh, J. M.</dc:creator>
<dc:date>2021-03-17</dc:date>
<dc:identifier>doi:10.1101/2021.03.15.435484</dc:identifier>
<dc:title><![CDATA[Functional development of a V3-specific broadly neutralizing antibody isolated from a case of HIV superinfection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.15.435527v1?rss=1">
<title>
<![CDATA[
Inflammatory signals are sufficient to elicit TOX expression in mouse and human CD8 T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.15.435527v1?rss=1</link>
<description><![CDATA[
T cell receptor (TCR) stimulation leads to expression of the transcription factor TOX. Prolonged TCR signaling, such as encountered during chronic infections or in tumors, leads to sustained TOX expression, which induces a state of exhaustion or dysfunction. While CD8 memory T cells (Tmem) in specific pathogen-free laboratory mice typically do not express TOX, functional human Tmem show heterogeneous TOX expression levels. Whether TCR-independent mechanisms can alter TOX expression in human and murine Tmem has not been defined. We report that human and mouse Tmem increase TOX expression following stimulation with inflammatory cytokines IL-12, IL-15, and IL-18. TOX and PD-1 expression patterns often appear to be directly correlated, however, we found that TOX is not necessary for cytokine-driven expression of PD-1. Together, these observations highlight that inflammation is sufficient to alter TOX and PD-1 expression and that the signals regulating TOX expression appear well conserved in human and murine Tmem.
]]></description>
<dc:creator>Maurice, N. J.</dc:creator>
<dc:creator>Berner, J.</dc:creator>
<dc:creator>Taber, A. K.</dc:creator>
<dc:creator>Zehn, D.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:date>2021-03-16</dc:date>
<dc:identifier>doi:10.1101/2021.03.15.435527</dc:identifier>
<dc:title><![CDATA[Inflammatory signals are sufficient to elicit TOX expression in mouse and human CD8 T cells]]></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/10.1101/2021.03.17.435863v1?rss=1">
<title>
<![CDATA[
Mutational escape from the polyclonal antibody response to SARS-CoV-2 infection is largely shaped by a single class of antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.17.435863v1?rss=1</link>
<description><![CDATA[
Monoclonal antibodies targeting a variety of epitopes have been isolated from individuals previously infected with SARS-CoV-2, but the relative contributions of these different antibody classes to the polyclonal response remains unclear. Here we use a yeast-display system to map all mutations to the viral spike receptor-binding domain (RBD) that escape binding by representatives of three potently neutralizing classes of anti-RBD antibodies with high-resolution structures. We compare the antibody-escape maps to similar maps for convalescent polyclonal plasma, including plasma from individuals from whom some of the antibodies were isolated. The plasma-escape maps most closely resemble those of a single class of antibodies that target an epitope on the RBD that includes site E484. Therefore, although the human immune system can produce antibodies that target diverse RBD epitopes, in practice the polyclonal response to infection is dominated by a single class of antibodies targeting an epitope that is already undergoing rapid evolution.
]]></description>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:creator>Weisblum, Y.</dc:creator>
<dc:creator>Schmidt, F.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Agudelo, M.</dc:creator>
<dc:creator>Finkin, S.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Muecksch, F.</dc:creator>
<dc:creator>Hatziioannou, T.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Robbiani, D. F.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-03-18</dc:date>
<dc:identifier>doi:10.1101/2021.03.17.435863</dc:identifier>
<dc:title><![CDATA[Mutational escape from the polyclonal antibody response to SARS-CoV-2 infection is largely shaped by a single class of antibodies]]></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/10.1101/2021.03.17.435909v1?rss=1">
<title>
<![CDATA[
High throughput genome-wide single cell protein:DNA binding site mapping by targeted insertion of promoters (TIP-seq) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.17.435909v1?rss=1</link>
<description><![CDATA[
Assessing cell to cell, and importantly, chromosome to chromosome, heterogeneity in cellular phenotypes is a central goal of modern cell biology. However, chromatin profiling in single cells has been extremely challenging, and single chromosome profiling has not been achieved. In cases where single cell methods have shown promise, success has been mainly limited to histone proteins and/or require highly specialized equipment or cell type specific protocols and are relatively low throughput. Here, we have combined the advantages of tagmentation, linear amplification and combinatorial indexing to produce a high throughput single cell DNA binding site mapping method that does not require specialized equipment and is capable of multiplexing several samples/ target proteins in one experiment. Targeted Insertion of Promoters (TIP-seq) uses Tn5 fused to protein A (as with CUT&Tag) to insert a T7 RNA polymerase promoter into sites adjacent to an antibody bound to a chromatin protein of interest, followed by linear amplification of flanking DNA with T7 polymerase, cDNA preparation and PCR indexing. Tip-seq provides [~]10-fold higher unique reads and thus higher coverage per single cell compared to state-of-the-art methods. We apply TIP-seq to map histone modifications, RNA PolII and CTCF binding sites in single human and mouse cells. TIP-seq will also be adaptable for other platforms, such as 10X genomics and ICELL8. In summary, TIP-seq provides a high-throughput, low-cost method for single cell protein mapping, that yields substantially higher coverage per cell and signal to noise than existing methods.
]]></description>
<dc:creator>Bartlett, D. A.</dc:creator>
<dc:creator>Dileep, V.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Gilbert, D. M.</dc:creator>
<dc:date>2021-03-19</dc:date>
<dc:identifier>doi:10.1101/2021.03.17.435909</dc:identifier>
<dc:title><![CDATA[High throughput genome-wide single cell protein:DNA binding site mapping by targeted insertion of promoters (TIP-seq)]]></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/10.1101/2021.03.22.436337v1?rss=1">
<title>
<![CDATA[
Live imaging of SARS-CoV-2 infection in mice reveals neutralizing antibodies require Fc function for optimal efficacy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.22.436337v1?rss=1</link>
<description><![CDATA[
Neutralizing antibodies (NAbs) are effective in treating COVID-19 but the mechanism of immune protection is not fully understood. Here, we applied live bioluminescence imaging (BLI) to monitor the real-time effects of NAb treatment in prophylaxis and therapy of K18-hACE2 mice intranasally infected with SARS-CoV-2-nanoluciferase. We could visualize virus spread sequentially from the nasal cavity to the lungs and thereafter systemically to various organs including the brain, which culminated in death. Highly potent NAbs from a COVID-19 convalescent subject prevented, and also effectively resolved, established infection when administered within three days. In addition to direct Fab-mediated neutralization, Fc effector interactions of NAbs with monocytes, neutrophils and natural killer cells were required to effectively dampen inflammatory responses and limit immunopathology. Our study highlights that both Fab and Fc effector functions of NAbs are essential for optimal in vivo efficacy against SARS-CoV-2.
]]></description>
<dc:creator>Ullah, I.</dc:creator>
<dc:creator>Prevost, J.</dc:creator>
<dc:creator>Ladinsky, M. S.</dc:creator>
<dc:creator>Stone, H.</dc:creator>
<dc:creator>Lu, M.</dc:creator>
<dc:creator>Anand, S. P.</dc:creator>
<dc:creator>Beaudoin-Bussieres, G.</dc:creator>
<dc:creator>Benlarbi, M.</dc:creator>
<dc:creator>Ding, S.</dc:creator>
<dc:creator>Gasser, R.</dc:creator>
<dc:creator>Fink, C.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Tauzin, A.</dc:creator>
<dc:creator>Goyette, G.</dc:creator>
<dc:creator>Bourassa, C.</dc:creator>
<dc:creator>Medjahed, H.</dc:creator>
<dc:creator>Mack, M.</dc:creator>
<dc:creator>Chung, K.</dc:creator>
<dc:creator>Wilen, C. B.</dc:creator>
<dc:creator>Dekaban, G. A.</dc:creator>
<dc:creator>Dikeakos, J. D.</dc:creator>
<dc:creator>Bruce, E. A.</dc:creator>
<dc:creator>Kaufmann, D. E.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>McGuire, A.</dc:creator>
<dc:creator>Richard, J.</dc:creator>
<dc:creator>Pazgier, M.</dc:creator>
<dc:creator>Bjorkman, P.</dc:creator>
<dc:creator>Mothes, W.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:creator>Kumar, P.</dc:creator>
<dc:creator>Uchil, P. D.</dc:creator>
<dc:date>2021-03-22</dc:date>
<dc:identifier>doi:10.1101/2021.03.22.436337</dc:identifier>
<dc:title><![CDATA[Live imaging of SARS-CoV-2 infection in mice reveals neutralizing antibodies require Fc function for optimal efficacy]]></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/10.1101/2021.03.23.436684v1?rss=1">
<title>
<![CDATA[
Isolation and Characterization of Cross-Neutralizing Coronavirus Antibodies from COVID-19+ Subjects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.23.436684v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 is one of three coronaviruses that have crossed the animal-to-human barrier in the past two decades. The development of a universal human coronavirus vaccine could prevent future pandemics. We characterized 198 antibodies isolated from four COVID19+ subjects and identified 14 SARS-CoV-2 neutralizing antibodies. One targeted the NTD, one recognized an epitope in S2 and twelve bound the RBD. Three anti-RBD neutralizing antibodies cross-neutralized SARS-CoV-1 by effectively blocking binding of both the SARS-CoV-1 and SARS-CoV-2 RBDs to the ACE2 receptor. Using the K18-hACE transgenic mouse model, we demonstrate that the neutralization potency rather than the antibody epitope specificity regulates the in vivo protective potential of anti-SARS-CoV-2 antibodies. The anti-S2 antibody also neutralized SARS-CoV-1 and all four cross-neutralizing antibodies neutralized the B.1.351 mutant strain. Thus, our study reveals that epitopes in S2 can serve as blueprints for the design of immunogens capable of eliciting cross-neutralizing coronavirus antibodies.
]]></description>
<dc:creator>Jennewein, M.</dc:creator>
<dc:creator>MacCamey, A.</dc:creator>
<dc:creator>Akins, N.</dc:creator>
<dc:creator>Feng, J.</dc:creator>
<dc:creator>Homad, L.</dc:creator>
<dc:creator>Hurlburt, N.</dc:creator>
<dc:creator>Seydoux, E.</dc:creator>
<dc:creator>Wang, Y.-H.</dc:creator>
<dc:creator>Stuart, A. B.</dc:creator>
<dc:creator>Edara, V. V.</dc:creator>
<dc:creator>Floyd, K.</dc:creator>
<dc:creator>Vanderheiden, A.</dc:creator>
<dc:creator>Mascola, J. R.</dc:creator>
<dc:creator>Doria-Rose, N.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Yang, E.</dc:creator>
<dc:creator>Chu, H.</dc:creator>
<dc:creator>Torres, J.</dc:creator>
<dc:creator>Ozorowski, G.</dc:creator>
<dc:creator>Ward, A.</dc:creator>
<dc:creator>Whaley, R.</dc:creator>
<dc:creator>Cohen, K.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>McElrath, J.</dc:creator>
<dc:creator>Englund, J. A.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:creator>Suthar, M.</dc:creator>
<dc:creator>McGuire, A.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:date>2021-03-24</dc:date>
<dc:identifier>doi:10.1101/2021.03.23.436684</dc:identifier>
<dc:title><![CDATA[Isolation and Characterization of Cross-Neutralizing Coronavirus Antibodies from COVID-19+ Subjects]]></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/10.1101/2021.04.06.438583v1?rss=1">
<title>
<![CDATA[
Highly-potent, synthetic APOBEC3s restrict HIV-1 through deamination-independent mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.06.438583v1?rss=1</link>
<description><![CDATA[
The APOBEC3 (A3) genes encode cytidine deaminase proteins with potent antiviral and anti-retroelement activity. This locus is characterized by duplication, recombination, and deletion events that gave rise to the seven A3s found in primates. These include three single deaminase domain A3s (A3A, A3C, and A3H) and four double deaminase domain A3s (A3B, A3D, A3F, and A3G). The most potent of the A3 proteins against HIV-1 is A3G. However, it is not clear if double deaminase domain A3s have a generalized functional advantage to restrict HIV-1. In order to test whether superior restriction factors could be created by genetically linking single A3 domains into synthetic double domains, we combined A3C and A3H single domains in novel combinations. We found that A3C/A3H double domains acquired enhanced antiviral activity that is at least as potent, if not better than, A3G. These synthetic double domain A3s have more efficiency of packaging into budding virions than their respective single domains, but this does not fully explain their gain of antiviral potency. The antiviral activity is conferred both by cytidine-deaminase dependent and independent mechanisms, with the latter correlating to an increase in RNA binding affinity. T cell lines expressing this A3C-A3H super restriction factor are able to control replicating HIV-1{Delta}Vif infection to similar levels as A3G. Together, these data show that novel combinations of A3 domains are capable of gaining potent antiviral activity to levels similar to the most potent genome-encoded A3s, via a primarily non-catalytic mechanism.

Author SummaryAntiviral genes are encoded by all organisms to help protect them from viral infections, including proteins encoded by primates to protect them from viruses similar to HIV-1. These antiviral proteins are also called "restriction factors". Some restriction factors are broadly acting, while others are very specific. During the course of evolution, some of these genes have expanded into multiple copies and rearranged in different versions to give them new activities. However, not all versions of these genes have been sampled in nature. In this paper, we validated the hypothesis that one particular antiviral gene family, called the APOBEC3 family, has the capability of making novel combinations of antiviral genes with as great, or greater, potency against HIV-1 as the most potent natural member of this family. By combining parts of the APOBEC3 proteins into novel combinations, we created potent antiviral versions that act through a mechanism distinct from existing APOBEC3 proteins.
]]></description>
<dc:creator>McDonnell, M. M.</dc:creator>
<dc:creator>Karvonen, S. C.</dc:creator>
<dc:creator>Gaba, A.</dc:creator>
<dc:creator>Flath, B.</dc:creator>
<dc:creator>Chelico, L.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2021-04-06</dc:date>
<dc:identifier>doi:10.1101/2021.04.06.438583</dc:identifier>
<dc:title><![CDATA[Highly-potent, synthetic APOBEC3s restrict HIV-1 through deamination-independent mechanisms]]></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/10.1101/2021.04.06.438709v1?rss=1">
<title>
<![CDATA[
Antibodies to the SARS-CoV-2 receptor-binding domain that maximize breadth and resistance to viral escape 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.06.438709v1?rss=1</link>
<description><![CDATA[
An ideal anti-SARS-CoV-2 antibody would resist viral escape1-3, have activity against diverse SARS-related coronaviruses4-7, and be highly protective through viral neutralization8-11 and effector functions12,13. Understanding how these properties relate to each other and vary across epitopes would aid development of antibody therapeutics and guide vaccine design. Here, we comprehensively characterize escape, breadth, and potency across a panel of SARS-CoV-2 antibodies targeting the receptor-binding domain (RBD), including S3094, the parental antibody of the late-stage clinical antibody VIR-7831. We observe a tradeoff between SARS-CoV-2 in vitro neutralization potency and breadth of binding across SARS-related coronaviruses. Nevertheless, we identify several neutralizing antibodies with exceptional breadth and resistance to escape, including a new antibody (S2H97) that binds with high affinity to all SARS-related coronavirus clades via a unique RBD epitope centered on residue E516. S2H97 and other escape-resistant antibodies have high binding affinity and target functionally constrained RBD residues. We find that antibodies targeting the ACE2 receptor binding motif (RBM) typically have poor breadth and are readily escaped by mutations despite high neutralization potency, but we identify one potent RBM antibody (S2E12) with breadth across sarbecoviruses closely related to SARS-CoV-2 and with a high barrier to viral escape. These data highlight functional diversity among antibodies targeting the RBD and identify epitopes and features to prioritize for antibody and vaccine development against the current and potential future pandemics.
]]></description>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Czudnochowski, N.</dc:creator>
<dc:creator>Zatta, F.</dc:creator>
<dc:creator>Park, Y.-J.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Pinto, D.</dc:creator>
<dc:creator>Beltramello, M.</dc:creator>
<dc:creator>Hernandez, P.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Marzi, R.</dc:creator>
<dc:creator>Glass, W. G.</dc:creator>
<dc:creator>Zhang, I.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Bowen, J. E.</dc:creator>
<dc:creator>Wojcechowskyj, J. A.</dc:creator>
<dc:creator>De Marco, A.</dc:creator>
<dc:creator>Rosen, L. E.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Montiel-Ruiz, M.</dc:creator>
<dc:creator>Kaiser, H.</dc:creator>
<dc:creator>Tucker, H.</dc:creator>
<dc:creator>Housley, M. P.</dc:creator>
<dc:creator>Di Iulio, J.</dc:creator>
<dc:creator>Lombardo, G.</dc:creator>
<dc:creator>Agostini, M.</dc:creator>
<dc:creator>Sprugasci, N.</dc:creator>
<dc:creator>Culap, K.</dc:creator>
<dc:creator>Jaconi, S.</dc:creator>
<dc:creator>Meury, M.</dc:creator>
<dc:creator>Dellota, E.</dc:creator>
<dc:creator>Cameroni, E.</dc:creator>
<dc:creator>Croll, T. I.</dc:creator>
<dc:creator>Nix, J. C.</dc:creator>
<dc:creator>Havenar-Daughton, C.</dc:creator>
<dc:creator>Telenti, A.</dc:creator>
<dc:creator>Lempp, F. A.</dc:creator>
<dc:creator>Pizzuto, M. S.</dc:creator>
<dc:creator>Chodera, J. D.</dc:creator>
<dc:creator>Hebner, C. M.</dc:creator>
<dc:creator>Whelan, S. P.</dc:creator>
<dc:creator>Virgin, H. W.</dc:creator>
<dc:creator>Veesler,</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.06.438709</dc:identifier>
<dc:title><![CDATA[Antibodies to the SARS-CoV-2 receptor-binding domain that maximize breadth and resistance to viral escape]]></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/10.1101/2021.04.07.438818v1?rss=1">
<title>
<![CDATA[
Structural basis for broad sarbecovirus neutralization by a human monoclonal antibody 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.07.438818v1?rss=1</link>
<description><![CDATA[
The recent emergence of SARS-CoV-2 variants of concern (VOC) and the recurrent spillovers of coronaviruses in the human population highlight the need for broadly neutralizing antibodies that are not affected by the ongoing antigenic drift and that can prevent or treat future zoonotic infections. Here, we describe a human monoclonal antibody (mAb), designated S2x259, recognizing a highly conserved cryptic receptor-binding domain (RBD) epitope and cross-reacting with spikes from all sarbecovirus clades. S2x259 broadly neutralizes spike-mediated entry of SARS-CoV-2 including the B.1.1.7, B.1.351, P.1 and B.1.427/B.1.429 VOC, as well as a wide spectrum of human and zoonotic sarbecoviruses through inhibition of ACE2 binding to the RBD. Furthermore, deep-mutational scanning and in vitro escape selection experiments demonstrate that S2x259 possesses a remarkably high barrier to the emergence of resistance mutants. We show that prophylactic administration of S2x259 protects Syrian hamsters against challenges with the prototypic SARS-CoV-2 and the B.1.351 variant, suggesting this mAb is a promising candidate for the prevention and treatment of emergent VOC and zoonotic infections. Our data unveil a key antigenic site targeted by broadly-neutralizing antibodies and will guide the design of pan-sarbecovirus vaccines.
]]></description>
<dc:creator>Tortorici, M. A.</dc:creator>
<dc:creator>Czudnochowski, N.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Marzi, R.</dc:creator>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>Zatta, F.</dc:creator>
<dc:creator>Bowen, J. E.</dc:creator>
<dc:creator>Jaconi, S.</dc:creator>
<dc:creator>di Iulio, J.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>De Marco, A.</dc:creator>
<dc:creator>Zepeda, S.</dc:creator>
<dc:creator>Pinto, D.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Beltramello, M.</dc:creator>
<dc:creator>Bartha, I.</dc:creator>
<dc:creator>Housley, M. P.</dc:creator>
<dc:creator>Lempp, F. A.</dc:creator>
<dc:creator>Rosen, L. E.</dc:creator>
<dc:creator>Dellota, E.</dc:creator>
<dc:creator>Kaiser, H.</dc:creator>
<dc:creator>Montiel-Ruiz, M.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Guarino, B.</dc:creator>
<dc:creator>Culap, K.</dc:creator>
<dc:creator>Sprugasci, N.</dc:creator>
<dc:creator>Saliba, C.</dc:creator>
<dc:creator>Vetti, E.</dc:creator>
<dc:creator>Giacchetto-Sasselli, I.</dc:creator>
<dc:creator>Silacci Fregni, C.</dc:creator>
<dc:creator>Abdelnabi, R.</dc:creator>
<dc:creator>Foo, C. S.-Y.</dc:creator>
<dc:creator>Havenar-Daughton, C.</dc:creator>
<dc:creator>Schmid, M. A.</dc:creator>
<dc:creator>Benigni, F.</dc:creator>
<dc:creator>Cameroni, E.</dc:creator>
<dc:creator>Neyts, J.</dc:creator>
<dc:creator>Telenti, A.</dc:creator>
<dc:creator>Snell, G.</dc:creator>
<dc:creator>Virgin, H. W.</dc:creator>
<dc:creator>Whelan, S. P. J.</dc:creator>
<dc:creator>Bloom, J.</dc:creator>
<dc:date>2021-04-08</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438818</dc:identifier>
<dc:title><![CDATA[Structural basis for broad sarbecovirus neutralization by a human monoclonal antibody]]></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/10.1101/2021.04.07.438871v1?rss=1">
<title>
<![CDATA[
Machine Learning Identifies Ponatinib as a Potent Inhibitor of SARS-CoV2-induced Cytokine Storm 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.07.438871v1?rss=1</link>
<description><![CDATA[
Although 15-20% of COVID-19 patients experience hyper-inflammation induced by massive cytokine production, cellular triggers of this process and strategies to target them remain poorly understood. Here, we show that the N-terminal domain (NTD) of the spike protein from the SARS-CoV-2 and emerging variants B1.1.7 and B.1.351 substantially induces multiple inflammatory molecules in human monocytes and PBMCs. Further, we identified several protein kinases, including JAK1, EPHA7, IRAK1, MAPK12, and MAP3K8, as essential downstream mediators of NTD-induced cytokine release. Additionally, we found that the FDA-approved, multi-kinase inhibitor Ponatinib is a potent inhibitor of the NTD-mediated cytokine storm. Taken together, we propose that agents targeting multiple kinases required for the SARS-CoV-2-mediated cytokine storm, such as Ponatinib, may represent an attractive therapeutic option for treating moderate to severe COVID-19.
]]></description>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Vijay, S.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:date>2021-04-09</dc:date>
<dc:identifier>doi:10.1101/2021.04.07.438871</dc:identifier>
<dc:title><![CDATA[Machine Learning Identifies Ponatinib as a Potent Inhibitor of SARS-CoV2-induced Cytokine Storm]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.09.439148v1?rss=1">
<title>
<![CDATA[
Structurally related but genetically unrelated antibody lineages converge on an immunodominantHIV-1 Env neutralizing determinant following trimer immunization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.09.439148v1?rss=1</link>
<description><![CDATA[
Understanding the molecular mechanisms by which antibodies target and neutralize the HIV-1 envelope glycoprotein (Env) is critical in guiding immunogen design and vaccine development aimed at eliciting cross-reactive neutralizing antibodies (NAbs). Here, we analyzed monoclonal antibodies (mAbs) isolated from non-human primates (NHPs) immunized with variants of a native flexibly linked (NFL) HIV-1 Env stabilized trimer derived from the tier 2 clade C 16055 strain. The antibodies displayed neutralizing activity against the autologous virus with potencies ranging from 0.005 to 3.68 ug/ml (IC50). Structural characterization using negative-stain EM and X-ray crystallography identified the variable region 2 (V2) of the 16055 NFL trimer to be the common epitope for these antibodies. The crystal structures revealed that the V2 segment adopts a {beta}-hairpin motif identical to that observed in the 16055 NFL crystal structure. These results depict how vaccine-induced antibodies derived from different clonal lineages penetrate through the glycan shield to recognize a hypervariable region within V2 (residues 184-186) that is unique to the 16055 strain. They also provide an explanation for the potent autologous neutralization of these antibodies, confirming the immunodominance of this site and revealing that multiple angles of approach are permissible for affinity/avidity that results in potent neutralizing capacity. The structural analysis reveals that the most negatively charged paratope correlated with the potency of the mAbs. The atomic level information is of interest to both define the means of autologous neutralization elicited by different tier 2-based immunogens and facilitate trimer redesign to better target more conserved regions of V2 to potentially elicit cross-neutralizing HIV-1 antibodies.

Author summaryNHPs immunizations with an HIV-1 immunogen (native-like tier 2 clade C 16055 strain) elicit potent HIV-1 tier 2 autologous polyclonal neutralizing antibodies. To understand the basis of the autologous neutralization, we determined structures of antibodies isolated from the vaccinated NHPs in complex with their epitopes. Our structural analysis reveals that the V2 hypervariable region, unique to 16055, is immunodominant and targeted by antibodies from diverse lineages. Additionally, vaccine-elicited V2 NAbs use different binding angles to avoid Env N-glycan shield and the more negatively charged paratope displays potent autologous neutralizing function. In summary, detailed analysis of how vaccine-elicited monoclonal antibodies interact with the target antigen provide valuable information for the design of immunogens aimed to elicit more broadly HIV-neutralizing antibodies. The use of cocktail/prime-boost sequential regimens that include a range of sequence variation combined with the removal/shielding of unwanted immunodominant epitopes will likely be needed to reach this goal.
]]></description>
<dc:creator>Aljedani, S.</dc:creator>
<dc:creator>Liban, T. J.</dc:creator>
<dc:creator>Tran, K.</dc:creator>
<dc:creator>Phad, G.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Dubrovskaya, V.</dc:creator>
<dc:creator>Pushparaj, P.</dc:creator>
<dc:creator>Martinez-Murillo, P.</dc:creator>
<dc:creator>Rodarte, J.</dc:creator>
<dc:creator>Mileant, A.</dc:creator>
<dc:creator>Mangala, V.</dc:creator>
<dc:creator>Kinzelman, R.</dc:creator>
<dc:creator>O'Dell, S.</dc:creator>
<dc:creator>Mascola, J. R.</dc:creator>
<dc:creator>Lee, K. K.</dc:creator>
<dc:creator>Karlsson Hedestam, G. B.</dc:creator>
<dc:creator>Wyatt, R. T.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:date>2021-04-09</dc:date>
<dc:identifier>doi:10.1101/2021.04.09.439148</dc:identifier>
<dc:title><![CDATA[Structurally related but genetically unrelated antibody lineages converge on an immunodominantHIV-1 Env neutralizing determinant following trimer immunization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.10.439265v1?rss=1">
<title>
<![CDATA[
Polypharmacological Re-programming of Tumor-associated Macrophages Restores Antitumor Immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.10.439265v1?rss=1</link>
<description><![CDATA[
Tumor-associated macrophages (TAMs) are an important component of the tumor microenvironment (TME), and they promote tumor progression, metastasis, and resistance to therapies. However, although TAMs represent a promising target for therapeutic intervention, the complexity of TME has made the study of TAMs challenging. Here, we established a physiologically-relevant in vitro TAM polarization system that recapitulates TAM pro-tumoral activities. We used this system for phenotypic kinase inhibitor screening and identified a multi-targeted compound BMS-794833 as the most potent inhibitor of TAM polarization. BMS-794833 decreased pro-tumoral properties of TAMs and suppressed tumor growth in mouse triple-negative breast cancer models. The effect of BMS-794833 was not dependent on its primary targets (MET and VEGFR2) but on its effect on multiple signaling pathways, including focal adhesion kinases, SRC family kinases, STAT3, and p38 MAP kinases. Our study underlines the efficacy of polypharmacological strategies in re-programming complex signaling cascades activated during TAM polarization.
]]></description>
<dc:creator>Nishida-Aoki, N.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:date>2021-04-11</dc:date>
<dc:identifier>doi:10.1101/2021.04.10.439265</dc:identifier>
<dc:title><![CDATA[Polypharmacological Re-programming of Tumor-associated Macrophages Restores Antitumor Immunity]]></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/10.1101/2021.04.13.439736v1?rss=1">
<title>
<![CDATA[
Multipotent progenitors and hematopoietic stem cells arise independently during the endothelial to hematopoietic transition in the early mouse embryo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.13.439736v1?rss=1</link>
<description><![CDATA[
During embryogenesis, waves of hematopoietic progenitors develop from hemogenic endothelium (HE) prior to the emergence of self-renewing hematopoietic stem cells (HSC). Although previous studies have shown that yolk sac-derived erythromyeloid progenitors and HSC emerge from distinct populations of HE, it remains unknown whether the earliest lymphoid-competent progenitors, multipotent progenitors, and HSC originate from common HE. Here we demonstrate by clonal assays and single cell transcriptomics that rare HE with functional HSC potential in the early murine embryo are distinct from more abundant HE with multilineage hematopoietic potential that fail to generate HSC. Specifically, HSC-competent HE are characterized by expression of CXCR4 surface marker and by higher expression of genes tied to arterial programs regulating HSC dormancy and self-renewal. Together, these findings suggest a revised model of developmental hematopoiesis in which the initial populations of multipotent progenitors and HSC arise independently from HE with distinct phenotypic and transcriptional properties.
]]></description>
<dc:creator>Dignum, T.</dc:creator>
<dc:creator>Varnum-Finney, B.</dc:creator>
<dc:creator>Srivatsan, S.</dc:creator>
<dc:creator>Dozono, S.</dc:creator>
<dc:creator>Waltner, O.</dc:creator>
<dc:creator>Heck, A.</dc:creator>
<dc:creator>Nourigat-McKay, C.</dc:creator>
<dc:creator>Jackson, D. L.</dc:creator>
<dc:creator>Rafii, S.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Bernstein, I. D.</dc:creator>
<dc:creator>Hadland, B. K.</dc:creator>
<dc:date>2021-04-14</dc:date>
<dc:identifier>doi:10.1101/2021.04.13.439736</dc:identifier>
<dc:title><![CDATA[Multipotent progenitors and hematopoietic stem cells arise independently during the endothelial to hematopoietic transition in the early mouse embryo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.14.439844v1?rss=1">
<title>
<![CDATA[
The SARS-CoV-2 mRNA-1273 vaccine elicits more RBD-focused neutralization, but with broader antibody binding within the RBD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.14.439844v1?rss=1</link>
<description><![CDATA[
The emergence of SARS-CoV-2 variants with mutations in key antibody epitopes has raised concerns that antigenic evolution will erode immunity. The susceptibility of immunity to viral evolution is shaped in part by the breadth of epitopes targeted. Here we compare the specificity of antibodies elicited by the mRNA-1273 vaccine versus natural infection. The neutralizing activity of vaccine-elicited antibodies is even more focused on the spike receptor-binding domain (RBD) than for infection-elicited antibodies. However, within the RBD, binding of vaccine-elicited antibodies is more broadly distributed across epitopes than for infection-elicited antibodies. This greater binding breadth means single RBD mutations have less impact on neutralization by vaccine sera than convalescent sera. Therefore, antibody immunity acquired by different means may have differing susceptibility to erosion by viral evolution.

One Sentence SummaryDeep mutational scanning shows the mRNA-1273 RBD-binding antibody response is less affected by single viral mutations than the infection response.
]]></description>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Gentles, L. E.</dc:creator>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Malone, K. D.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-04-14</dc:date>
<dc:identifier>doi:10.1101/2021.04.14.439844</dc:identifier>
<dc:title><![CDATA[The SARS-CoV-2 mRNA-1273 vaccine elicits more RBD-focused neutralization, but with broader antibody binding within the RBD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.16.440249v1?rss=1">
<title>
<![CDATA[
Genomic Stability and Genetic Defense Systems in Dolosigranulum pigrum a Candidate Beneficial Bacterium from the Human Microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.16.440249v1?rss=1</link>
<description><![CDATA[
Dolosigranulum pigrum is positively associated with indicators of health in multiple epidemiological studies of human nasal microbiota. Knowledge of the basic biology of D. pigrum is a prerequisite for evaluating its potential for future therapeutic use; however, such data are very limited. To gain insight into D. pigrums chromosomal structure, pangenome and genomic stability, we compared the genomes of 28 D. pigrum strains that were collected across 20 years. Phylogenomic analysis showed closely related strains circulating over this period and closure of 19 genomes revealed highly conserved chromosomal synteny. Gene clusters involved in the mobilome and in defense against mobile genetic elements (MGEs) were enriched in the accessory genome versus the core genome. A systematic analysis for MGEs identified the first candidate D. pigrum prophage and insertion sequence. A systematic analysis for genetic elements that limit the spread of MGEs, including restriction modification (RM), CRISPR-Cas, and deity-named defense systems, revealed strain-level diversity in host defense systems that localized to specific genomic sites including one RM system hotspot. Analysis of CRISPR spacers pointed to a wealth of MGEs against which D. pigrum defends itself. These results reveal a role for horizontal gene transfer and mobile genetic elements in strain diversification while highlighting that in D. pigrum this occurs within the context of a highly stable chromosomal organization protected by a variety of defense mechanisms.

IMPORTANCEDolosigranulum pigrum is a candidate beneficial bacterium with potential for future therapeutic use. This is based on its positive associations with characteristics of health in multiple studies of human nasal microbiota across the span of human life. For example, high levels of D. pigrum nasal colonization in adults predicts the absence of Staphylococcus aureus nasal colonization. Also, D. pigrum nasal colonization in young children is associated with healthy control groups in studies of middle ear infections. Our analysis of 28 genomes revealed a remarkable stability of D. pigrum strains colonizing people in the U.S. across a 20-year span. We subsequently identified factors that can influence this stability, including genomic stability, phage predators, the role of MGEs in strain-level variation and defenses against MGEs. Finally, these D. pigrum strains also lacked predicted virulence factors. Overall, these findings add additional support to the potential for D. pigrum as a therapeutic bacterium.
]]></description>
<dc:creator>Flores Ramos, S.</dc:creator>
<dc:creator>Brugger, S. D.</dc:creator>
<dc:creator>Fernandez Escapa, I.</dc:creator>
<dc:creator>Skeete, C. A.</dc:creator>
<dc:creator>Cotton, S. L.</dc:creator>
<dc:creator>Eslami, S. M.</dc:creator>
<dc:creator>Gao, W.</dc:creator>
<dc:creator>Bomar, L.</dc:creator>
<dc:creator>Tran, T. H.</dc:creator>
<dc:creator>Jones, D. S.</dc:creator>
<dc:creator>Minot, S.</dc:creator>
<dc:creator>Roberts, R. J.</dc:creator>
<dc:creator>Johnston, C. D.</dc:creator>
<dc:creator>Lemon, K. P.</dc:creator>
<dc:date>2021-04-18</dc:date>
<dc:identifier>doi:10.1101/2021.04.16.440249</dc:identifier>
<dc:title><![CDATA[Genomic Stability and Genetic Defense Systems in Dolosigranulum pigrum a Candidate Beneficial Bacterium from the Human Microbiome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.20.440612v1?rss=1">
<title>
<![CDATA[
Redefining replication in multi-ancestry genome-wide association studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.20.440612v1?rss=1</link>
<description><![CDATA[
Since 2005, genome-wide association (GWA) datasets have been largely biased toward sampling European ancestry individuals, and recent studies have shown that GWA results estimated from self-identified European individuals are not transferable to non-European individuals due to various confounding challenges. Here, we demonstrate that enrichment analyses which aggregate SNP-level association statistics at multiple genomic scales--from genes to genomic regions and pathways--have been underutilized in the GWA era and can generate biologically interpretable hypotheses regarding the genetic basis of complex trait architecture. We illustrate examples of the robust associations generated by enrichment analyses while studying 25 continuous traits assayed in 566,786 individuals from seven diverse self-identified human ancestries in the UK Biobank and the Biobank Japan, as well as 44,348 admixed individuals from the PAGE consortium including cohorts of African-American, Hispanic and Latin American, Native Hawaiian, and American Indian/Alaska Native individuals. We identify 1,000 gene-level associations that are genome-wide significant in at least two ancestry cohorts across these 25 traits, as well as highly conserved pathway associations with triglyceride levels in European, East Asian, and Native Hawaiian cohorts.
]]></description>
<dc:creator>Smith, S. P.</dc:creator>
<dc:creator>Shahamatdar, S.</dc:creator>
<dc:creator>Cheng, W.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Paik, J.</dc:creator>
<dc:creator>Graff, M.</dc:creator>
<dc:creator>Haiman, C. A.</dc:creator>
<dc:creator>Matise, T. C.</dc:creator>
<dc:creator>North, K. E.</dc:creator>
<dc:creator>Peters, U.</dc:creator>
<dc:creator>Kenny, E.</dc:creator>
<dc:creator>Gignoux, C. R.</dc:creator>
<dc:creator>Wojcik, G. L.</dc:creator>
<dc:creator>Crawford, L.</dc:creator>
<dc:creator>Ramachandran, S.</dc:creator>
<dc:date>2021-04-20</dc:date>
<dc:identifier>doi:10.1101/2021.04.20.440612</dc:identifier>
<dc:title><![CDATA[Redefining replication in multi-ancestry genome-wide association studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.20.440672v1?rss=1">
<title>
<![CDATA[
BET family members Bdf1/2 modulate global transcription initiation and elongation in Saccharomyces cerevisiae 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.20.440672v1?rss=1</link>
<description><![CDATA[
Human bromodomain-containing BET family members are promising targets for therapy of cancer and immunoinflammatory diseases, but their mechanisms of action and functional redundancies are poorly understood. Bdf1/2, yeast homologues of the human BET factors, were previously proposed to target transcription factor TFIID to acetylated histone H4, analogous to bromodomains that are present within the largest subunit of metazoan TFIID. We investigated the genome-wide roles of Bdf1/2 and found that their important contributions to transcription extend beyond TFIID function, as transcription of many genes is more sensitive to Bdf1/2 than to TFIID depletion. Bdf1/2 co-occupy the majority of yeast promoters and affect preinitiation complex formation through recruitment of TFIID, Mediator and basal transcription factors to chromatin. Surprisingly, we discovered that hypersensitivity of genes to Bdf1/2 depletion results from combined defects in transcription initiation and processive elongation, a striking functional similarity to human BET proteins, most notably Brd4. Our results establish Bdf1/2 as critical for yeast transcription and provide important mechanistic insights into the function of BET proteins in all eukaryotes.
]]></description>
<dc:creator>Donczew, R.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:date>2021-04-20</dc:date>
<dc:identifier>doi:10.1101/2021.04.20.440672</dc:identifier>
<dc:title><![CDATA[BET family members Bdf1/2 modulate global transcription initiation and elongation in Saccharomyces cerevisiae]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.22.441020v1?rss=1">
<title>
<![CDATA[
Multi-omics profiling of U2AF1 mutants dissects pathogenic mechanisms affecting RNA granules in myeloid malignancies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.22.441020v1?rss=1</link>
<description><![CDATA[
Somatic mutations in splicing factors are of significant interest in myeloid malignancies and other cancers. U2AF1, together with U2AF2, is essential for 3 splice site recognition. U2AF1 mutations result in aberrant splicing, but the molecular mechanism and the full spectrum of consequences on RNA biology have not been fully elucidated to date. We performed multi-omics profiling of in vivo RNA binding, splicing and turnover for U2AF1 S34F and Q157R mutants. We dissected specific binding signals of U2AF1 and U2AF2 and showed that U2AF1 mutations individually alter U2AF1-RNA binding, resulting in defective U2AF2 recruitment. We demonstrated a complex relationship between differential binding and splicing, expanding upon the currently accepted loss-of-binding model. Finally, we observed that U2AF1 mutations increase the formation of stress granules in both cell lines and primary acute myeloid leukemia samples. Our results uncover U2AF1 mutation-dependent pathogenic RNA mechanisms and provide the basis for developing targeted therapeutic strategies.
]]></description>
<dc:creator>Biancon, G.</dc:creator>
<dc:creator>Joshi, P.</dc:creator>
<dc:creator>Zimmer, J. T.</dc:creator>
<dc:creator>Hunck, T.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Lessard, M. D.</dc:creator>
<dc:creator>Courchaine, E.</dc:creator>
<dc:creator>Barentine, A. E.</dc:creator>
<dc:creator>Machyna, M.</dc:creator>
<dc:creator>Botti, V.</dc:creator>
<dc:creator>Qin, A.</dc:creator>
<dc:creator>Gbyli, R.</dc:creator>
<dc:creator>Patel, A.</dc:creator>
<dc:creator>Song, Y.</dc:creator>
<dc:creator>Kiefer, L.</dc:creator>
<dc:creator>Viero, G.</dc:creator>
<dc:creator>Neuenkirchen, N.</dc:creator>
<dc:creator>Lin, H.</dc:creator>
<dc:creator>Bewersdorf, J.</dc:creator>
<dc:creator>Simon, M. D.</dc:creator>
<dc:creator>Neugebauer, K. M.</dc:creator>
<dc:creator>Tebaldi, T.</dc:creator>
<dc:creator>Halene, S.</dc:creator>
<dc:date>2021-04-23</dc:date>
<dc:identifier>doi:10.1101/2021.04.22.441020</dc:identifier>
<dc:title><![CDATA[Multi-omics profiling of U2AF1 mutants dissects pathogenic mechanisms affecting RNA granules in myeloid malignancies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.25.441339v1?rss=1">
<title>
<![CDATA[
Kinetochore-associated Mps1 regulates the strength of kinetochore-microtubule attachments via Ndc80 phosphorylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.25.441339v1?rss=1</link>
<description><![CDATA[
Dividing cells detect and correct erroneous kinetochore-microtubule attachments during mitosis, thereby avoiding chromosome mis-segregation. Most studies of this process have focused on the Aurora B kinase, which phosphorylates microtubule-binding elements specifically at incorrectly attached kinetochores, promoting their release and providing another chance for proper attachments to form. However, growing evidence suggests additional mechanisms, potentially involving Mps1 kinase, that also underlie error correction. Because these mechanisms overlap in vivo, and because both Mps1 and Aurora B function in numerous other vital processes, their contributions to the correction of erroneous kinetochore attachments have been difficult to disentangle. Here we directly examine how Mps1 activity affects kinetochore-microtubule attachments using a reconstitution-based approach that allowed us to separate its effects from Aurora B activity. When endogenous Mps1 that co-purifies with isolated kinetochores is activated in vitro, it weakens their attachments to microtubules via phosphorylation of Ndc80, a major microtubule-binding element of the outer kinetochore. Mps1 phosphorylation of Ndc80 appears to contribute to error correction because phospho-deficient Ndc80 mutants exhibit genetic interactions and segregation defects when combined with mutants in an intrinsic error correction pathway. In addition, Mps1 phosphorylation of Ndc80 is stimulated on kinetochores lacking tension. These data suggest that Mps1 provides an additional mechanism for correcting erroneous kinetochore-microtubule attachments, complementing the well-known activity of Aurora B.
]]></description>
<dc:creator>Sarangapani, K. K.</dc:creator>
<dc:creator>Koch, L. B.</dc:creator>
<dc:creator>Nelson, C. R.</dc:creator>
<dc:creator>Asbury, C. L.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2021-04-26</dc:date>
<dc:identifier>doi:10.1101/2021.04.25.441339</dc:identifier>
<dc:title><![CDATA[Kinetochore-associated Mps1 regulates the strength of kinetochore-microtubule attachments via Ndc80 phosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-26</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.

O_FIG O_LINKSMALLFIG WIDTH=182 HEIGHT=200 SRC="FIGDIR/small/441344v1_ufig1.gif" ALT="Figure 1">
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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/10.1101/2021.05.06.442916v1?rss=1">
<title>
<![CDATA[
Identification of DAXX As A Restriction Factor Of SARS-CoV-2 Through A CRISPR/Cas9 Screen 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.06.442916v1?rss=1</link>
<description><![CDATA[
Interferon restricts SARS-CoV-2 replication in cell culture, but only a handful of Interferon Stimulated Genes with antiviral activity against SARS-CoV-2 have been identified. Here, we describe a functional CRISPR/Cas9 screen aiming at identifying SARS-CoV-2 restriction factors. We identified DAXX, a scaffold protein residing in PML nuclear bodies known to limit the replication of DNA viruses and retroviruses, as a potent inhibitor of SARS-CoV-2 and SARS-CoV replication in human cells. Basal expression of DAXX was sufficient to limit the replication of SARS-CoV-2, and DAXX over-expression further restricted infection. In contrast with most of its previously described antiviral activities, DAXX-mediated restriction of SARS-CoV-2 was independent of the SUMOylation pathway. SARS-CoV-2 infection triggered the re-localization of DAXX to cytoplasmic sites and promoted its degradation. Mechanistically, this process was mediated by the viral papain-like protease (PLpro) and the proteasome. Together, these results demonstrate that DAXX restricts SARS-CoV-2, which in turn has evolved a mechanism to counteract its action.
]]></description>
<dc:creator>Mac Kain, A.</dc:creator>
<dc:creator>Maarifi, G.</dc:creator>
<dc:creator>Aicher, S.-M.</dc:creator>
<dc:creator>Arhel, N.</dc:creator>
<dc:creator>Baidaliuk, A.</dc:creator>
<dc:creator>Vallet, T.</dc:creator>
<dc:creator>Tran, Q. D.</dc:creator>
<dc:creator>Hardy, A.</dc:creator>
<dc:creator>Chazal, M.</dc:creator>
<dc:creator>Porrot, F.</dc:creator>
<dc:creator>Ohainle, M.</dc:creator>
<dc:creator>Carlson-Stevermer, J.</dc:creator>
<dc:creator>Oki, J.</dc:creator>
<dc:creator>Holden, K.</dc:creator>
<dc:creator>Simon-Loriere, E.</dc:creator>
<dc:creator>Bruel, T.</dc:creator>
<dc:creator>Schwartz, O.</dc:creator>
<dc:creator>Jouvenet, N.</dc:creator>
<dc:creator>Nisole, S.</dc:creator>
<dc:creator>Vignuzzi, M.</dc:creator>
<dc:creator>Roesch, F.</dc:creator>
<dc:date>2021-05-06</dc:date>
<dc:identifier>doi:10.1101/2021.05.06.442916</dc:identifier>
<dc:title><![CDATA[Identification of DAXX As A Restriction Factor Of SARS-CoV-2 Through A CRISPR/Cas9 Screen]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.10.443350v1?rss=1">
<title>
<![CDATA[
Individual Level Differential Expression Analysis for Single Cell RNA-seq data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.10.443350v1?rss=1</link>
<description><![CDATA[
Bulk RNA-seq data quantify the expression of a gene in an individual by one number (e.g., fragment count). In contrast, single cell RNA-seq (scRNA-seq) data provide much richer information: the distribution of gene expression across many cells. To assess differential expression across individuals using scRNA-seq data, a straightforward solution is to create "pseudo" bulk RNA-seq data by adding up the fragment counts of a gene across cells for each individual, and then apply methods designed for differential expression using bulk RNA-seq data. This pseudo-bulk solution reduces the distribution of gene expression across cells to a single number and thus loses a good amount of information. We propose to assess differential expression using the gene expression distribution measured by cell level data. We find denoising cell level data can substantially improve the power of this approach. We apply our method, named IDEAS (Individual level Differential Expression Analysis for scRNA-seq), to study the gene expression difference between autism subjects and controls. We find neurogranin-expressing neurons harbor a high proportion of differentially expressed genes, and ERBB signals in microglia are associated with autism.
]]></description>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Miao, Z.</dc:creator>
<dc:creator>Han, F.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2021-05-10</dc:date>
<dc:identifier>doi:10.1101/2021.05.10.443350</dc:identifier>
<dc:title><![CDATA[Individual Level Differential Expression Analysis for Single Cell RNA-seq data]]></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/10.1101/2021.05.10.443452v1?rss=1">
<title>
<![CDATA[
Therapeutic reversal of prenatal pontine ID1 signaling in DIPG 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.10.443452v1?rss=1</link>
<description><![CDATA[
Diffuse intrinsic pontine glioma (DIPG) is a highly aggressive brain tumor with rare survival beyond two years. This poor prognosis is largely due to the tumors highly infiltrative and invasive nature. Previous reports demonstrate upregulation of the transcription factor ID1 with H3K27M and ACVR1 mutations, but this has not been confirmed in human tumors or therapeutically targeted. We developed an in utero electroporation (IUE) murine H3K27M-driven tumor model, which demonstrates increased ID1 expression in H3K27M- and ACVR1-mutated tumor cells. In human tumors, elevated ID1 expression is associated with H3K27M/ACVR1-mutation, brainstem location, and reduced survival. The ID1 promoter demonstrates a similar active epigenetic state in H3K27M tumor cells and murine prenatal hindbrain cells. In the developing human brain, ID1 is expressed highest in oligo/astrocyte-precursor cells (OAPCs). These ID1+/SPARCL1+ cells share a transcriptional program with astrocyte-like (AC-like) DIPG cells, and demonstrate upregulation of gene sets involved with regulation of cell migration. Both genetic and pharmacologic [cannabidiol (CBD)] suppression of ID1 results in decreased DIPG cell invasion/migration in vitro and invasion/tumor growth in multiple in vivo models. CBD reduces proliferation through reactive oxygen species (ROS) production at low micromolar concentrations, which we found to be achievable in the murine brainstem. Further, pediatric high-grade glioma patients treated off-trial with CBD (n=15) demonstrate tumor ID1 reduction and improved overall survival compared to historical controls. Our study identifies that ID1 is upregulated in DIPG through reactivation of a developmental OAPC transcriptional state, and ID1-driven invasiveness of DIPG is therapeutically targetable with CBD.

One Sentence SummaryThe transcription factor ID1 is upregulated in a subset of DIPG tumor cells, and ID1-driven invasiveness is therapeutically targetable with CBD.
]]></description>
<dc:creator>Yadav, V. N.</dc:creator>
<dc:creator>Harris, M. K.</dc:creator>
<dc:creator>Messinger, D.</dc:creator>
<dc:creator>Thomas, C.</dc:creator>
<dc:creator>Cummings, J. R.</dc:creator>
<dc:creator>Yang, T.</dc:creator>
<dc:creator>Woo, R.</dc:creator>
<dc:creator>Siddaway, R.</dc:creator>
<dc:creator>Burkert, M.</dc:creator>
<dc:creator>Stallard, S.</dc:creator>
<dc:creator>Qin, T.</dc:creator>
<dc:creator>Mullan, B.</dc:creator>
<dc:creator>Siada, R.</dc:creator>
<dc:creator>Ravindran, R.</dc:creator>
<dc:creator>Niculcea, M.</dc:creator>
<dc:creator>Ginn, K.</dc:creator>
<dc:creator>Gener, M. A. H.</dc:creator>
<dc:creator>Dorris, K.</dc:creator>
<dc:creator>Vitanza, N. A.</dc:creator>
<dc:creator>Schmidt, S.</dc:creator>
<dc:creator>Spitzer, J.</dc:creator>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>Filbin, M.</dc:creator>
<dc:creator>Cao, X.</dc:creator>
<dc:creator>Castro, M.</dc:creator>
<dc:creator>Lowenstein, P.</dc:creator>
<dc:creator>Mody, R.</dc:creator>
<dc:creator>Chinnaiyan, A.</dc:creator>
<dc:creator>Desprez, P.-Y.</dc:creator>
<dc:creator>McAllister, S.</dc:creator>
<dc:creator>Hawkins, C.</dc:creator>
<dc:creator>Waszak, S.</dc:creator>
<dc:creator>Venneti, S.</dc:creator>
<dc:creator>Koschmann, C.</dc:creator>
<dc:date>2021-05-11</dc:date>
<dc:identifier>doi:10.1101/2021.05.10.443452</dc:identifier>
<dc:title><![CDATA[Therapeutic reversal of prenatal pontine ID1 signaling in DIPG]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-11</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/10.1101/2021.05.11.443605v1?rss=1">
<title>
<![CDATA[
Comprehensive profiling of mutations to influenza virus PB2 that confer resistance to the cap-binding inhibitor pimodivir 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.11.443605v1?rss=1</link>
<description><![CDATA[
Antivirals are used not only in current treatment of influenza, but are also stockpiled as a first line of defense against novel influenza strains for which vaccines have yet to be developed. Identifying drug resistance mutations can guide clinical deployment of the antiviral, and additionally define the mechanisms of drug action and drug resistance. Pimodivir is a first-in-class inhibitor of the polymerase basic protein 2 (PB2) subunit of the influenza A virus polymerase complex. A number of resistance mutations have previously been identified in treated patients or cell culture. Here, we generate a complete map of the effect of all single-amino-acid mutations to an avian PB2 on resistance to pimodivir. We identified both known and novel resistance mutations not only in the previously implicated cap-binding and mid-link domains, but also in the N-terminal domain. Our complete map of pimodivir resistance thus enables the evaluation of whether new viral strains contain mutations that will confer pimodivir resistance.
]]></description>
<dc:creator>Soh, Y. Q. S.</dc:creator>
<dc:creator>Malone, K. D.</dc:creator>
<dc:creator>Eguia, R. T.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-05-11</dc:date>
<dc:identifier>doi:10.1101/2021.05.11.443605</dc:identifier>
<dc:title><![CDATA[Comprehensive profiling of mutations to influenza virus PB2 that confer resistance to the cap-binding inhibitor pimodivir]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.11.443665v1?rss=1">
<title>
<![CDATA[
A graphical, interactive and GPU-enabled workflow to process long-read sequencing data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.11.443665v1?rss=1</link>
<description><![CDATA[
We present a graphical cloud-enabled workflow for fast, interactive analysis of nanopore sequencing data using GPUs. Users customize parameters, monitor execution and visualize results through an accessible graphical interface. To facilitate reproducible deployment, we use Docker containers and provide an Amazon Machine Image (AMI) with all software and drivers pre-installed for GPU computing on the cloud. We observe a 34x speedup and a 109x reduction in costs for the rate-limiting basecalling step in the analysis of blood cancer cell line data. The graphical interface and greatly simplified deployment facilitate the adoption of GPUs for rapid, cost-effective analysis of long-read sequencing.
]]></description>
<dc:creator>Reddy, S.</dc:creator>
<dc:creator>Hung, L.-H.</dc:creator>
<dc:creator>Sala-Torra, O.</dc:creator>
<dc:creator>Radich, J.</dc:creator>
<dc:creator>Yeung, C. C.</dc:creator>
<dc:creator>Yeung, K. Y.</dc:creator>
<dc:date>2021-05-13</dc:date>
<dc:identifier>doi:10.1101/2021.05.11.443665</dc:identifier>
<dc:title><![CDATA[A graphical, interactive and GPU-enabled workflow to process long-read sequencing data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.12.443914v1?rss=1">
<title>
<![CDATA[
Chromosome length and gene density contribute to micronuclear membrane stability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.12.443914v1?rss=1</link>
<description><![CDATA[
Micronuclei are derived from missegregated chromosomes and frequently lose membrane integrity, leading to DNA damage, innate immune activation, and metastatic signaling. Here we demonstrate that two characteristics of the trapped chromosome, length and gene density, are key contributors to micronuclei membrane stability in human cells. Chromosome length is proportional to micronuclei size, and gene density has an additive effect with micronucleus size on membrane stability. We demonstrate that these results are not due to chromosome-specific differences in spindle position or initial nuclear pore complex recruitment during post-mitotic nuclear envelope assembly. We find that chromosome length and micronuclei size strongly correlate with lamin B1 and nuclear pore density in intact micronuclei. Unexpectedly, lamin B1 levels do not predict nuclear lamina organization and membrane stability. Instead, small gene-dense micronuclei have decreased nuclear lamina gaps compared to large micronuclei, despite very low levels of lamin B1. Our data strongly suggest that nuclear envelope composition defects previously correlated with membrane rupture only partly explain membrane stability in micronuclei. We propose that an unknown factor linked to gene density has a separate function that inhibits the appearance of nuclear lamina gaps and delays membrane rupture until late in the cell cycle.
]]></description>
<dc:creator>Mammel, A.</dc:creator>
<dc:creator>Huang, H. Z.</dc:creator>
<dc:creator>Gunn, A. L.</dc:creator>
<dc:creator>Choo, E.</dc:creator>
<dc:creator>Hatch, E. M.</dc:creator>
<dc:date>2021-05-14</dc:date>
<dc:identifier>doi:10.1101/2021.05.12.443914</dc:identifier>
<dc:title><![CDATA[Chromosome length and gene density contribute to micronuclear membrane stability]]></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/10.1101/2021.05.15.444222v1?rss=1">
<title>
<![CDATA[
Stabilization of the SARS-CoV-2 Spike receptor-binding domain using deep mutational scanning and structure-based design 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.15.444222v1?rss=1</link>
<description><![CDATA[
The unprecedented global demand for SARS-CoV-2 vaccines has demonstrated the need for highly effective vaccine candidates that are thermostable and amenable to large-scale manufacturing. Nanoparticle immunogens presenting the receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein (S) in repetitive arrays are being advanced as second-generation vaccine candidates, as they feature robust manufacturing characteristics and have shown promising immunogenicity in preclinical models. Here, we used previously reported deep mutational scanning (DMS) data to guide the design of stabilized variants of the RBD. The selected mutations fill a cavity in the RBD that has been identified as a linoleic acid binding pocket. Screening of several designs led to the selection of two lead candidates that expressed at higher yields than the wild-type RBD. These stabilized RBDs possess enhanced thermal stability and resistance to aggregation, particularly when incorporated into an icosahedral nanoparticle immunogen that maintained its integrity and antigenicity for 28 days at 35-40{degrees}C, while corresponding immunogens displaying the wild-type RBD experienced aggregation and loss of antigenicity. The stabilized immunogens preserved the potent immunogenicity of the original nanoparticle immunogen, which is currently being evaluated in a Phase I/II clinical trial. Our findings may improve the scalability and stability of RBD-based coronavirus vaccines in any format and more generally highlight the utility of comprehensive DMS data in guiding vaccine design.
]]></description>
<dc:creator>Ellis, D.</dc:creator>
<dc:creator>Brunette, N.</dc:creator>
<dc:creator>Crawford, K. H. D.</dc:creator>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>Pham, M. N.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Herpoldt, K.-L.</dc:creator>
<dc:creator>Fiala, B.</dc:creator>
<dc:creator>Murphy, M.</dc:creator>
<dc:creator>Pettie, D.</dc:creator>
<dc:creator>Kraft, J. C.</dc:creator>
<dc:creator>Malone, K. D.</dc:creator>
<dc:creator>Navarro, M. J.</dc:creator>
<dc:creator>Ogohara, C.</dc:creator>
<dc:creator>Kepl, E.</dc:creator>
<dc:creator>Ravichandran, R.</dc:creator>
<dc:creator>Sydeman, C.</dc:creator>
<dc:creator>Ahlrichs, M.</dc:creator>
<dc:creator>Johnson, M.</dc:creator>
<dc:creator>Blackstone, A.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Lee, K. K.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:date>2021-05-16</dc:date>
<dc:identifier>doi:10.1101/2021.05.15.444222</dc:identifier>
<dc:title><![CDATA[Stabilization of the SARS-CoV-2 Spike receptor-binding domain using deep mutational scanning and structure-based design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.17.444326v1?rss=1">
<title>
<![CDATA[
Patients with mesenchymal tumours and high Fusobacteriales prevalence have worse prognosis in colorectal cancer (CRC) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.17.444326v1?rss=1</link>
<description><![CDATA[
ObjectiveTranscriptomic-based subtyping, Consensus Molecular Subtyping (CMS) and CRC Intrinsic Subtyping (CRIS), identify a patient subpopulation with mesenchymal traits (CMS4/CRIS-B) and poorer outcome. Here, we investigated the relationship between prevalence of Fusobacterium nucleatum (Fn) and Fusobacteriales, CMS/CRIS subtyping, cell type composition, immune infiltrates and host contexture to refine patients stratification and identify druggable context-specific vulnerabilities.

DesignWe coupled cell culture experiments with characterization of Fn/Fusobacteriales prevalence and host biology/microenviroment in tumours from 2 independent CRC patient cohorts (Taxonomy: n=140; TCGA-COAD-READ: n=605).

ResultsIn vitro, Fn infection induced inflammation via NF{kappa}B/TNF in HCT116 and HT29 cancer cell lines. In patients, high Fn/Fusobacteriales were found in CMS1, MSI tumours, with infiltration of macrophages M1, reduced macrophages M2, and high IL6/IL8/IL1{beta} signaling. Analysis of the Taxonomy cohort suggested that Fn was prognostic for CMS4/CRIS-B patients, despite having lower Fn load than CMS1 patients. In the TCGA-COAD-READ cohort, we likewise identified a differential association between Fusobacteriales relative abundance and outcome when stratifying patients in mesenchymal (either CMS4 and/or CRIS-B) vs. non-mesenchymal (neither CMS4 nor CRIS-B). Patients with mesenchymal tumours and high Fusobacteriales had approximately 2-fold higher risk of worse outcome. These associations were null in non-mesenchymal patients. Modelling the 3-way association between Fusobacteriales prevalence, molecular subtyping, and host contexture with logistic models with an interaction term disentangled the pathogen/host-signaling relationship and identified aberrations (including EMT/WNT/NOTCH) as candidate targets.

ConclusionThis study identifies CMS4/CRIS-B patients with high Fn/Fusobacteriales prevalence as a high-risk subpopulation that may benefit from therapeutics targeting mesenchymal biology.

Significance of this studyO_ST_ABSWhat is already known on this subject?C_ST_ABSO_LIFusobacterium nucleatum (Fn), a commensal Gram-negative anaerobe from the Fusobacteriales order, is an onco-bacterium in CRC as a causal relationship between Fn prevalence and CRC pathogenesis, progression and treatment response has been reported in vivo.
C_LIO_LIBroad spectrum antibiotics has proven moderately successful in reducing tumour growth in preclinical models. However, the use of antibiotics to treat bacterium-positive cases in the clinic is not a viable option as it may further alter the already dysbiotic gut microbiome of CRC patients and may also have limited efficacy against Fn which penetrates and embeds deeply within the tumour.
C_LIO_LIThe highly heterogenous CRC patient population can be classified into distinct molecular subtypes (CMS and CRIS) based on gene expression profiles mirroring the underlying transcriptional programs. Patients classified as CMS4 and CRIS-B exhibit a mesenchymal phenotype and have poorer outcome.
C_LI

What are the new findings?O_LIFn/Fusobacteriales prevalence is associated with immune involvement (decrease in macrophages M1 and increase in macrophages M2) and activation of specific signalling programs (inflammation, DNA damage, WNT, metastasis, proliferation, cell cycle) in the host tumours.
C_LIO_LIThe prevalence of bacteria from the Fusobacteriales order, largely driven by Fn species, play an active or opportunistic role depending on the underlying host tumour biology and microenvironment.
C_LIO_LIFn and other species of the Fusobacteriales order are enriched in CMS1 (immuno, microsatellite unstable) patients compared to CMS2-4 cases.
C_LIO_LIFn/Fusobacteriales prevalence is associated with worse clinical outcome in patients with mesenchymal-rich CMS4/CRIS-B tumours, but not in patients with other molecular subtypes.
C_LI

How might it impact on clinical practice in the foreseeable future?O_LIFn/Fusobacteriales screening and transcriptomic-based molecular subtyping should be considered to identify patients with mesenchymal-rich tumours and high bacterium prevalence and to inform disease management.
C_LIO_LIFn/Fusobacteriales prevalence may need to be addressed exclusively in patients with mesenchymal-rich high-stromal infiltrating tumours rather than a blanket-approach to treat all pathogen-positive patients.
C_LIO_LIClinical management of the disease for this subpopulation of high-risk patients with unfavourable clinical outcome could be attained by administering compounds currently in clinical trials that target aberrations in the host signaling pathways (NOTCH, WNT, EMT) and tumour microenviroment (inflammasome, activated T cells, complement system, and macrophage chemotactism and activation).
C_LI
]]></description>
<dc:creator>Salvucci, M.</dc:creator>
<dc:creator>Crawford, N. T.</dc:creator>
<dc:creator>Stott, K.</dc:creator>
<dc:creator>Bullman, S.</dc:creator>
<dc:creator>Longley, D. B.</dc:creator>
<dc:creator>Prehn, J. H.</dc:creator>
<dc:date>2021-05-18</dc:date>
<dc:identifier>doi:10.1101/2021.05.17.444326</dc:identifier>
<dc:title><![CDATA[Patients with mesenchymal tumours and high Fusobacteriales prevalence have worse prognosis in colorectal cancer (CRC)]]></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/10.1101/2021.05.20.445000v1?rss=1">
<title>
<![CDATA[
Functional dissection of human mitotic proteins using CRISPR-Cas9 tiling screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.20.445000v1?rss=1</link>
<description><![CDATA[
Kinetochores are large protein complexes that assemble at the centromere and bind to mitotic spindle microtubules to ensure accurate chromosome segregation. Like most protein-coding genes, the full multifunctional nature of kinetochore factors remains uncharacterized due to the limited experimental tools for unbiased dissection of human protein sequences. We developed a method that leverages CRISPR-Cas9 induced mutations to identify key functional regions within protein sequences required for cellular outgrowth. Our analysis of 48 human mitotic genes revealed hundreds of regions required for cell proliferation, including known domains and uncharacterized ones. We validated the essential nature for 15 of these regions, including amino acids 387-402 of Mad1, which identified an unknown domain that contributes to Mad1 kinetochore localization and chromosome segregation fidelity. Altogether, we demonstrate that CRISPR-Cas9-based tiling mutagenesis identifies key functional domains in protein-coding genes de novo, which elucidates separation of function mutants and allows functional annotation across the human proteome.
]]></description>
<dc:creator>Herman, J. A.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:creator>Paddison, P.</dc:creator>
<dc:date>2021-05-20</dc:date>
<dc:identifier>doi:10.1101/2021.05.20.445000</dc:identifier>
<dc:title><![CDATA[Functional dissection of human mitotic proteins using CRISPR-Cas9 tiling screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.20.445014v1?rss=1">
<title>
<![CDATA[
Superscan: Supervised Single-Cell Annotation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.20.445014v1?rss=1</link>
<description><![CDATA[
Automated cell type annotation of single-cell RNA-seq data has the potential to significantly improve and streamline single cell data analysis, facilitating comparisons and meta-analyses. However, many of the current state-of-the-art techniques suffer from limitations, such as reliance on a single reference dataset or marker gene set, or excessive run times for large datasets. Acquiring high-quality labeled data to use as a reference can be challenging. With CITE-seq, surface protein expression of cells can be directly measured in addition to the RNA expression, facilitating cell type annotation. Here, we compiled and annotated a collection of 16 publicly available CITE-seq datasets. This data was then used as training data to develop Superscan, a supervised machine learning-based prediction model. Using our 16 reference datasets, we benchmarked Superscan and showed that it performs better in terms of both accuracy and speed when compared to other state-of-the-art cell annotation methods. Superscan is pre-trained on a collection of primarily PBMC immune datasets; however, additional data and cell types can be easily added to the training data for further improvement. Finally, we used Superscan to reanalyze a previously published dataset, demonstrating its applicability even when the dataset includes cell types that are missing from the training set.
]]></description>
<dc:creator>Shasha, C.</dc:creator>
<dc:creator>Tian, Y.</dc:creator>
<dc:creator>Mair, F.</dc:creator>
<dc:creator>Miller, H. E. R.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:date>2021-05-22</dc:date>
<dc:identifier>doi:10.1101/2021.05.20.445014</dc:identifier>
<dc:title><![CDATA[Superscan: Supervised Single-Cell Annotation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.21.445198v1?rss=1">
<title>
<![CDATA[
Transcriptional and Functional Activity of Canine Hemangiosarcoma to Support Hematopoiesis Demonstrate Bone Marrow Nurse Cell Ontogeny 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.21.445198v1?rss=1</link>
<description><![CDATA[
Hemangiosarcoma and angiosarcoma are soft-tissue sarcomas of blood vessel-forming cells in dogs and humans, respectively. These vasoformative sarcomas are aggressive and highly metastatic, with disorganized, irregular blood-filled vascular spaces. Our objective was to define molecular programs which support the niche that enables progression of canine hemangiosarcoma and human angiosarcoma. Dog-in-mouse hemangiosarcoma xenografts recapitulated the vasoformative and highly angiogenic morphology and molecular characteristics of primary tumors. Blood vessels in the tumors were complex and disorganized, and they were lined by both donor and host cells, a trait that was not observed in xenografts from canine osteosarcoma and lymphoma. In some cases, the xenografted hemangiosarcoma cells created exuberant myeloid hyperplasia and gave rise to lymphoproliferative tumors of mouse origin. We did not uncover a definitive transmissible etiology, but our functional analyses indicate that hemangiosarcoma cells generate a microenvironment that supports expansion and differentiation of hematopoietic progenitor populations. We conclude that canine hemangiosarcomas, and possibly human angiosarcomas, originate from stromal cells that are part of the bone marrow niche and that these cells may also support the growth of hematopoietic tumors.

SignificanceWe demonstrate that molecular programs supporting expansion of immune and inflammatory cells in hemangiosarcoma resemble those of bone marrow niche cells, providing insights into the potential roles of these cells - whether physiological or pathological - in creating a permissive environment for the progression of hematopoietic malignancies.
]]></description>
<dc:creator>Kim, J. H.</dc:creator>
<dc:creator>Schulte, A. J.</dc:creator>
<dc:creator>Sarver, A. L.</dc:creator>
<dc:creator>Angelos, M. G.</dc:creator>
<dc:creator>Frantz, A. M.</dc:creator>
<dc:creator>Foster, C. L.</dc:creator>
<dc:creator>O'Brien, T. D.</dc:creator>
<dc:creator>Cornax, I.</dc:creator>
<dc:creator>O'Sullivan, M. G.</dc:creator>
<dc:creator>Cheng, N.</dc:creator>
<dc:creator>Lewellen, M.</dc:creator>
<dc:creator>Oseth, L.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Bullman, S.</dc:creator>
<dc:creator>Pedamallu, C. S.</dc:creator>
<dc:creator>Goyal, S. M.</dc:creator>
<dc:creator>Meyerson, M.</dc:creator>
<dc:creator>Lund, T. C.</dc:creator>
<dc:creator>Alfoldi, J.</dc:creator>
<dc:creator>Lindblad-Toh, K.</dc:creator>
<dc:creator>Breen, M.</dc:creator>
<dc:creator>Dickerson, E. B.</dc:creator>
<dc:creator>Kaufman, D. S.</dc:creator>
<dc:creator>Modiano, J. F.</dc:creator>
<dc:date>2021-05-23</dc:date>
<dc:identifier>doi:10.1101/2021.05.21.445198</dc:identifier>
<dc:title><![CDATA[Transcriptional and Functional Activity of Canine Hemangiosarcoma to Support Hematopoiesis Demonstrate Bone Marrow Nurse Cell Ontogeny]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.21.445204v1?rss=1">
<title>
<![CDATA[
Met is required for oligodendrocyte progenitor cell migration in Danio rerio 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.21.445204v1?rss=1</link>
<description><![CDATA[
During vertebrate central nervous system development, most oligodendrocyte progenitor cells (OPCs) are specified in the ventral spinal cord and must migrate throughout the neural tube until they become evenly distributed, occupying non-overlapping domains. While this process of developmental OPC migration is well characterized, the nature of the molecular mediators that govern it remain largely unknown. Here, using zebrafish as a model, we demonstrate that Met signaling is required for initial developmental migration of OPCs, and, using cell-specific knock-down of Met signaling, show that Met acts cell-autonomously in OPCs. Taken together, these findings demonstrate in vivo, the role of Met signaling in OPC migration and provide new insight into how OPC migration is regulated during development.
]]></description>
<dc:creator>Ali, M. F.</dc:creator>
<dc:creator>Latimer, A. J.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Hogenmiller, L.</dc:creator>
<dc:creator>Fontenas, L.</dc:creator>
<dc:creator>Isabella, A. J.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:creator>Yu, G.</dc:creator>
<dc:creator>Kucenas, S.</dc:creator>
<dc:date>2021-05-23</dc:date>
<dc:identifier>doi:10.1101/2021.05.21.445204</dc:identifier>
<dc:title><![CDATA[Met is required for oligodendrocyte progenitor cell migration in Danio rerio]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.24.445486v1?rss=1">
<title>
<![CDATA[
Nonsense-mediated mRNA decay utilizes complementary mechanisms to suppress mRNA and protein accumulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.24.445486v1?rss=1</link>
<description><![CDATA[
Nonsense-mediated mRNA decay (NMD) is an essential, highly conserved quality control pathway that detects and degrades mRNAs containing premature termination codons (PTCs). Although the essentiality of NMD is frequently ascribed to its prevention of truncated protein accumulation, the extent to which NMD actually suppresses proteins encoded by NMD-sensitive transcripts is less well-understood than NMD-mediated suppression of mRNA. Here, we describe a reporter system that permits accurate quantification of both mRNA and protein levels via stable integration of paired reporters encoding NMD-sensitive and NMD-insensitive transcripts into the AAVS1 safe harbor loci in human cells. We use this system to demonstrate that NMD suppresses proteins encoded by NMD-sensitive transcripts by up to [~]8-fold more than the mRNA itself. Our data indicate that NMD limits the accumulation of proteins encoded by NMD substrates by mechanisms beyond mRNA degradation, such that even when NMD-sensitive mRNAs escape destruction, their encoded proteins are still effectively suppressed.
]]></description>
<dc:creator>Udy, D. B.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:date>2021-05-25</dc:date>
<dc:identifier>doi:10.1101/2021.05.24.445486</dc:identifier>
<dc:title><![CDATA[Nonsense-mediated mRNA decay utilizes complementary mechanisms to suppress mRNA and protein accumulation]]></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/10.1101/2021.05.26.442666v1?rss=1">
<title>
<![CDATA[
Longitudinal immune dynamics of mild COVID-19 define signatures of recovery and persistence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.26.442666v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 has infected over 200 million and caused more than 4 million deaths to date. Most individuals (>80%) have mild symptoms and recover in the outpatient setting, but detailed studies of immune responses have focused primarily on moderate to severe COVID-19. We deeply profiled the longitudinal immune response in individuals with mild COVID-19 beginning with early time points post-infection (1-15 days) and proceeding through convalescence to >100 days after symptom onset. We correlated data from single cell analyses of peripheral blood cells, serum proteomics, virus-specific cellular and humoral immune responses, and clinical metadata. Acute infection was characterized by vigorous coordinated innate and adaptive immune activation that differed in character by age (young vs. old). We then characterized signals associated with recovery and convalescence to define and validate a new signature of inflammatory cytokines, gene expression, and chromatin accessibility that persists in individuals with post-acute sequelae of SARS-CoV-2 infection (PASC).
]]></description>
<dc:creator>Talla, A.</dc:creator>
<dc:creator>Vasaikar, S. V.</dc:creator>
<dc:creator>Lemos, M. P.</dc:creator>
<dc:creator>Moodie, Z.</dc:creator>
<dc:creator>Pebworth, M.-P. L.</dc:creator>
<dc:creator>Henderson, K. E.</dc:creator>
<dc:creator>Cohen, K. W.</dc:creator>
<dc:creator>Czartoski, J. L.</dc:creator>
<dc:creator>Lai, L.</dc:creator>
<dc:creator>Suthar, M. S.</dc:creator>
<dc:creator>Heubeck, A. T.</dc:creator>
<dc:creator>Genge, P. C.</dc:creator>
<dc:creator>Roll, C. M.</dc:creator>
<dc:creator>Weiss, M.</dc:creator>
<dc:creator>Reading, J.</dc:creator>
<dc:creator>Kondza, N.</dc:creator>
<dc:creator>MacMillan, H.</dc:creator>
<dc:creator>Fong, O. C.</dc:creator>
<dc:creator>Thomson, Z. J.</dc:creator>
<dc:creator>Graybuck, L. T.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Coffey, E. M.</dc:creator>
<dc:creator>Meijer, P.</dc:creator>
<dc:creator>Becker, L. A.</dc:creator>
<dc:creator>De Rosa, S. C.</dc:creator>
<dc:creator>Torgerson, T. R.</dc:creator>
<dc:creator>Skene, P. J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Szeto, G. L.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Bumol, T. F.</dc:creator>
<dc:date>2021-05-26</dc:date>
<dc:identifier>doi:10.1101/2021.05.26.442666</dc:identifier>
<dc:title><![CDATA[Longitudinal immune dynamics of mild COVID-19 define signatures of recovery and persistence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.01.446671v1?rss=1">
<title>
<![CDATA[
Identifying strengths and weaknesses of methods for computational network inference from single cell RNA-seq data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.01.446671v1?rss=1</link>
<description><![CDATA[
Single-cell RNA-sequencing (scRNA-seq) offers unparalleled insight into the transcriptional programs of different cellular states by measuring the transcriptome of thousands of individual cells. An emerging problem in the analysis of scRNA-seq is the inference of transcriptional gene regulatory networks and a number of methods with different learning frameworks have been developed to address this problem. Here, we present an expanded benchmarking study of eleven recent network inference methods on six published scRNA-seq datasets in human, mouse, and yeast considering different types of gold standard networks and evaluation metrics. We evaluate methods based on their computing requirements as well as on their ability to recover the network structure. We find that, while most methods have a modest recovery of experimentally derived interactions based on global metrics such as Area Under the Precision Recall curve, methods are able to capture targets of regulators that are relevant to the system under study. Among the top performing methods that use only expression were SCENIC, PIDC, MERLIN or Correlation. Addition of prior biological knowledge and the estimation of transcription factor activities resulted in the best overall performance with the Inferelator and MERLIN methods that use prior knowledge outperforming methods that use expression alone. We found little to no benefit from imputation for network inference, which is further dataset-dependent. Comparisons of inferred networks for comparable bulk conditions showed that the networks inferred from scRNA-seq datasets are often better or at par with the networks inferred from bulk datasets. Our analysis should be beneficial in selecting methods for network inference. At the same time, this highlights the need for improved methods and better gold standards for regulatory network inference from scRNAseq datasets.
]]></description>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>McCalla, S. G.</dc:creator>
<dc:creator>Fotuhi Siahpirani, A.</dc:creator>
<dc:creator>Periyasamy, V.</dc:creator>
<dc:creator>Shin, J.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:date>2021-06-02</dc:date>
<dc:identifier>doi:10.1101/2021.06.01.446671</dc:identifier>
<dc:title><![CDATA[Identifying strengths and weaknesses of methods for computational network inference from single cell RNA-seq data]]></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/10.1101/2021.06.07.447395v1?rss=1">
<title>
<![CDATA[
T helper 2 transcriptional profile predicts single-cell HIV envelope-specific polyfunctional CD4+ T cells correlated with reduced risk of infection in RV144 trial 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.07.447395v1?rss=1</link>
<description><![CDATA[
Despite the critical role antigen-specific T cells play in responding to viral infections, their aggregate frequencies in peripheral blood have not correlated with clinical protection during HIV infection. However, a subset of HIV-specific CD4+ T cells, termed polyfunctional T cells, can produce multiple effector cytokines simultaneously. In the RV144 HIV vaccine trial, polyfunctional T cells correlated with reduced risk of HIV infection. Little is known about what differentiates polyfunctional T cells from other vaccine-elicited T cells. Therefore, we developed a novel live-cell multiplexed cytokine capture assay, to identify and transcriptionally profile vaccine-specific polyfunctional CD4+ T cells. We applied these methods to samples from the HVTN 097 clinical trial of the same vaccine regimen as RV144. We discovered two surface receptors that were enriched among polyfunctional CD4+ T cells and a Th2-biased signature (IL-4, IL-5, and IL-13) that specifically predicted the envelope-specific polyfunctional CD4+ T cells that were correlated with reduced risk of HIV infection in RV144. By linking single-cell transcriptional and functional profiles, we may be able to further define the role of vaccine-elicited polyfunctional T cells in contributing to effective immunity.

Key PointsO_LINovel ex vivo multiplexed cytokine capture assay to enumerate and single-cell sort polyfunctional T cells for downstream analyses
C_LIO_LIPolyfunctional T cells were specifically detected among the HIV envelope-stimulated CD4+ T cells
C_LIO_LISingle-cell RNA sequencing identified novel surface markers enriched among vaccine-specific polyfunctional CD4+ T cells
C_LIO_LITh2 transcriptional signature predicted polyfunctional CD4+ T cell profile that had correlated with reduced risk of HIV infection in the RV144 HIV efficacy trial
C_LI
]]></description>
<dc:creator>Cohen, K. W.</dc:creator>
<dc:creator>Tian, Y.</dc:creator>
<dc:creator>Thayer, C.</dc:creator>
<dc:creator>Seese, A.</dc:creator>
<dc:creator>Amezquita, R.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>De Rosa, S.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:date>2021-06-07</dc:date>
<dc:identifier>doi:10.1101/2021.06.07.447395</dc:identifier>
<dc:title><![CDATA[T helper 2 transcriptional profile predicts single-cell HIV envelope-specific polyfunctional CD4+ T cells correlated with reduced risk of infection in RV144 trial]]></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/10.1101/2021.06.14.041913v1?rss=1">
<title>
<![CDATA[
The atypical RNA-binding protein TAF15 regulates dorsoanterior neural development through diverse mechanisms in Xenopus tropicalis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.14.041913v1?rss=1</link>
<description><![CDATA[
The FET family of atypical RNA-binding proteins includes Fused in sarcoma (Fus), Ewings sarcoma (EWS), and the TATA-binding protein-associate factor 15 (TAF15). FET proteins are highly conserved, suggesting specialized requirements for each protein. Fus regulates splicing of transcripts required for mesoderm differentiation and cell adhesion in Xenopus, but roles that EWS and TAF15 play remain unknown. Here we analyze the roles of maternally deposited and zygotically transcribed TAF15, which is essential for the proper development of dorsoanterior neural tissues. By measuring changes in exon usage and transcript abundance from TAF15-depleted embryos we found TAF15 may regulate dorsoanterior neural development through fgfr4 and ventx2.1. TAF15 uses distinct mechanisms to downregulate FGFR4 expression: 1) retention of a single intron within fgfr4 when maternal and zygotic TAF15 is depleted, and 2) reduction of total fgfr4 transcript when zygotic TAF15 alone is depleted. The two mechanisms of gene regulation (post-transcriptional vs transcriptional) suggest TAF15-mediated gene regulation is target and cofactor-dependent, depending on the milieu of factors that are present at different times of development.
]]></description>
<dc:creator>DeJong, C. S.</dc:creator>
<dc:creator>Dichmann, D. S.</dc:creator>
<dc:creator>Exner, C. R. T.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Harland, R. M.</dc:creator>
<dc:date>2021-06-14</dc:date>
<dc:identifier>doi:10.1101/2021.06.14.041913</dc:identifier>
<dc:title><![CDATA[The atypical RNA-binding protein TAF15 regulates dorsoanterior neural development through diverse mechanisms in Xenopus tropicalis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.15.448495v1?rss=1">
<title>
<![CDATA[
Tracking SARS-CoV-2 Spike Protein Mutations in the United States (2020/01 - 2021/03) Using a Statistical Learning Strategy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.15.448495v1?rss=1</link>
<description><![CDATA[
The emergence and establishment of SARS-CoV-2 variants of interest (VOI) and variants of concern (VOC) highlight the importance of genomic surveillance. We propose a statistical learning strategy (SLS) for identifying and spatiotemporally tracking potentially relevant Spike protein mutations. We analyzed 167,893 Spike protein sequences from US COVID-19 cases (excluding 21,391 sequences from VOI/VOC strains) deposited at GISAID from January 19, 2020 to March 15, 2021. Alignment against the reference Spike protein sequence led to the identification of viral residue variants (VRVs), i.e., residues harboring a substitution compared to the reference strain. Next, generalized additive models were applied to model VRV temporal dynamics, to identify VRVs with significant and substantial dynamics (false discovery rate q-value <0.01; maximum VRV proportion > 10% on at least one day).

Unsupervised learning was then applied to hierarchically organize VRVs by spatiotemporal patterns and identify VRV-haplotypes. Finally, homology modelling was performed to gain insight into potential impact of VRVs on Spike protein structure. We identified 90 VRVs, 71 of which have not previously been observed in a VOI/VOC, and 35 of which have emerged recently and are durably present. Our analysis identifies 17 VRVs [~]91 days earlier than their first corresponding VOI/VOC publication. Unsupervised learning revealed eight VRV-haplotypes of 4 VRVs or more, suggesting two emerging strains (B1.1.222 and B.1.234). Structural modeling supported potential functional impact of the D1118H and L452R mutations. The SLS approach equally monitors all Spike residues over time, independently of existing phylogenic classifications, and is complementary to existing genomic surveillance methods.
]]></description>
<dc:creator>Zhao, L. P.</dc:creator>
<dc:creator>Lybrand, T. P.</dc:creator>
<dc:creator>Gilbert, P. B.</dc:creator>
<dc:creator>Hawn, T. R.</dc:creator>
<dc:creator>Schiffer, J. T.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Payne, T. H.</dc:creator>
<dc:creator>Carpp, L.</dc:creator>
<dc:creator>Geraghty, D. E.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:date>2021-06-15</dc:date>
<dc:identifier>doi:10.1101/2021.06.15.448495</dc:identifier>
<dc:title><![CDATA[Tracking SARS-CoV-2 Spike Protein Mutations in the United States (2020/01 - 2021/03) Using a Statistical Learning Strategy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.18.449051v1?rss=1">
<title>
<![CDATA[
Recovery of deleted deep sequencing data sheds more light on the early Wuhan SARS-CoV-2 epidemic 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.18.449051v1?rss=1</link>
<description><![CDATA[
The origin and early spread of SARS-CoV-2 remains shrouded in mystery. Here I identify a data set containing SARS-CoV-2 sequences from early in the Wuhan epidemic that has been deleted from the NIHs Sequence Read Archive. I recover the deleted files from the Google Cloud, and reconstruct partial sequences of 13 early epidemic viruses. Phylogenetic analysis of these sequences in the context of carefully annotated existing data further supports the idea that the Huanan Seafood Market sequences are not fully representative of the viruses in Wuhan early in the epidemic. Instead, the progenitor of currently known SARS-CoV-2 sequences likely contained three mutations relative to the market viruses that made it more similar to SARS-CoV-2s bat coronavirus relatives.
]]></description>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-06-22</dc:date>
<dc:identifier>doi:10.1101/2021.06.18.449051</dc:identifier>
<dc:title><![CDATA[Recovery of deleted deep sequencing data sheds more light on the early Wuhan SARS-CoV-2 epidemic]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.02.450872v1?rss=1">
<title>
<![CDATA[
Mammalian UPF3A and UPF3B activate NMD independently of their EJC binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.02.450872v1?rss=1</link>
<description><![CDATA[
Nonsense-mediated mRNA decay (NMD) is governed by the three conserved factors - UPF1, UPF2 and UPF3. While all three are required for NMD in yeast, UPF3B is dispensable for NMD in mammals, with its paralog UPF3A suggested to only weakly activate or even repress NMD due to its weaker binding to the exon junction complex (EJC). Here we characterize the UPF3B-dependent and -independent NMD in human cell lines knocked-out of one or both UPF3 paralogs. We show that in human colorectal cancer HCT116 cells, EJC-mediated NMD can operate in UPF3B-dependent and -independent manner. While UPF3A is almost completely dispensable for NMD in wild-type cells, it strongly activates EJC-mediated NMD in cells lacking UPF3B. Surprisingly, this major NMD branch can operate in UPF3-independent manner questioning the idea that UPF3 is needed to bridge UPF proteins to the EJC during NMD. Complementation studies in UPF3 knockout cells further show that EJC-binding domain of UPF3 paralogs is not essential for NMD. Instead, the conserved mid domain of UPF3B, previously shown to engage with ribosome release factors, is required for its full NMD activity. Altogether, UPF3 plays a more active role in NMD than simply being a bridge between the EJC and the UPF complex.
]]></description>
<dc:creator>Yi, Z.</dc:creator>
<dc:creator>Arvola, R. M.</dc:creator>
<dc:creator>Myers, S.</dc:creator>
<dc:creator>Dilsavor, C. N.</dc:creator>
<dc:creator>Abu Alhasan, R.</dc:creator>
<dc:creator>Carter, B. N.</dc:creator>
<dc:creator>Patton, R. D.</dc:creator>
<dc:creator>Bundschuh, R.</dc:creator>
<dc:creator>Singh, G.</dc:creator>
<dc:date>2021-07-03</dc:date>
<dc:identifier>doi:10.1101/2021.07.02.450872</dc:identifier>
<dc:title><![CDATA[Mammalian UPF3A and UPF3B activate NMD independently of their EJC binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.03.450733v1?rss=1">
<title>
<![CDATA[
Combined tumor and immune signals from genomes or transcriptomes predict outcomes of checkpoint inhibition in melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.03.450733v1?rss=1</link>
<description><![CDATA[
Cancer immunotherapy with checkpoint blockade (CPB) leads to improved outcomes in melanoma and other tumor types, but a majority of patients do not respond. High tumor mutation burden (TMB) and high levels of tumor-infiltrating T cells have been associated with response to immunotherapy, but integrative models to predict clinical benefit using DNA or RNA alone have not been comprehensively explored. We sequenced DNA and RNA from melanoma patients receiving CPB, and aggregated previously published data, yielding whole exome sequencing data for 189 patients and bulk RNA sequencing data for 178 patients. Using these datasets, we derived genomic and transcriptomic factors that predict overall survival (OS) and response to immunotherapy. Using whole-exome DNA data alone, we calculated T cell burden (TCB) and B cell burden (BCB) based on rearranged TCR/Ig DNA sequences and found that patients whose melanomas have high TMB together with either high TCB or high BCB survived longer and had higher response rates as compared to patients with either low TMB or TCB/BCB. Next, using bulk RNA-Seq data, differential expression analysis identified 83 genes associated with high or low OS. By combining pairs of immune-expressed genes with tumor-expressed genes, we identified three gene pairs associated with response and survival (Bonferroni P<0.05). All three gene pair models were validated in an independent cohort (n=180) (Bonferroni P<0.05). The best performing gene pair model included the lymphocyte-expressed MAP4K1 (Mitogen- Activated Protein Kinase Kinase Kinase Kinase 1) combined with the transcription factor TBX3 (T-Box Transcription Factor 3) which is overexpressed in poorly differentiated melanomas. We conclude that RNA-based (MAP4K1&TBX3) or DNA-based (TCB&TMB) models combining immune and tumor measures improve predictions of outcome after checkpoint blockade in melanoma.
]]></description>
<dc:creator>Freeman, S. S.</dc:creator>
<dc:creator>Sade-Feldman, M.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Stewart, C.</dc:creator>
<dc:creator>Gonye, A. L. K.</dc:creator>
<dc:creator>Ravi, A.</dc:creator>
<dc:creator>Arniella, M. B.</dc:creator>
<dc:creator>Gushterova, I.</dc:creator>
<dc:creator>LaSalle, T. J.</dc:creator>
<dc:creator>Blaum, E. M.</dc:creator>
<dc:creator>Yizhak, K.</dc:creator>
<dc:creator>Frederick, D. T.</dc:creator>
<dc:creator>Sharova, T.</dc:creator>
<dc:creator>Leshchiner, I.</dc:creator>
<dc:creator>Elagina, L.</dc:creator>
<dc:creator>Spiro, O. G.</dc:creator>
<dc:creator>Livitz, D.</dc:creator>
<dc:creator>Rosebrock, D.</dc:creator>
<dc:creator>Aguet, F.</dc:creator>
<dc:creator>Carrot-Zhang, J.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Chen, J. H.</dc:creator>
<dc:creator>Barzily-Rokni, M.</dc:creator>
<dc:creator>Hammond, M. R.</dc:creator>
<dc:creator>Vitzthum von Eckstaedt, H. C.</dc:creator>
<dc:creator>Blackmon, S. M.</dc:creator>
<dc:creator>Jiao, Y. J.</dc:creator>
<dc:creator>Gabriel, S.</dc:creator>
<dc:creator>Lawrence, D. P.</dc:creator>
<dc:creator>Duncan, L. M.</dc:creator>
<dc:creator>Stemmer-Rachamimov, A. O.</dc:creator>
<dc:creator>Wargo, J. A.</dc:creator>
<dc:creator>Flaherty, K. T.</dc:creator>
<dc:creator>Sullivan, R. J.</dc:creator>
<dc:creator>Boland, G. M.</dc:creator>
<dc:creator>Meyerson, M.</dc:creator>
<dc:creator>Getz, G.</dc:creator>
<dc:creator>Hacohen, N.</dc:creator>
<dc:date>2021-07-04</dc:date>
<dc:identifier>doi:10.1101/2021.07.03.450733</dc:identifier>
<dc:title><![CDATA[Combined tumor and immune signals from genomes or transcriptomes predict outcomes of checkpoint inhibition in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.03.450987v1?rss=1">
<title>
<![CDATA[
eQTL mapping using allele-specific gene expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.03.450987v1?rss=1</link>
<description><![CDATA[
Using information from allele-specific gene expression (ASE) can sub-stantially improve the power to map gene expression quantitative trait loci (eQTLs). However, such practice has been limited, partly due to high computational cost and the requirement to access raw data that can take a large amount of storage space. To address these computational challenges, we have developed a computational framework that uses a statistical method named TReCASE as its computational engine, and it is computationally feasible for large scale analysis. We applied it to map eQTLs in 28 human tissues using the data from the Genotype-Tissue Expression (GTEx) project. Compared with a popular linear regression method that does not use ASE data, TReCASE can double the number of eGenes (i.e., genes with at least one significant eQTL) when sample size is relatively small, e.g., n = 200. We also demonstrated how to use the ASE data that we have collected to study dynamic eQTLs whose effect sizes vary with respect to another variable, such as age. We find the majority of such dynamic eQTLs are due to some underlying latent factors, such as cell type proportions. We further compare TReCASE versus another method RASQUAL. TReCASE is ten times or more faster than RASQUAL and it provides more robust type I error control.
]]></description>
<dc:creator>Zhabotynsky, V.</dc:creator>
<dc:creator>Huang, L.</dc:creator>
<dc:creator>Little, P.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Pardo Manuel de Villena, F. F.</dc:creator>
<dc:creator>Zou, F.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2021-07-04</dc:date>
<dc:identifier>doi:10.1101/2021.07.03.450987</dc:identifier>
<dc:title><![CDATA[eQTL mapping using allele-specific gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.04.450352v1?rss=1">
<title>
<![CDATA[
PHENOTYPIC CHARACTERIZATION OF TWO NOVEL CELL LINE MODELS OF CASTRATION RESISTANT PROSTATE CANCER. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.04.450352v1?rss=1</link>
<description><![CDATA[
BACKGROUNDResistance to androgen deprivation therapies is a major driver of mortality in advanced prostate cancer. Therefore, there is a need to develop new pre-clinical models that allow the investigation of resistance mechanisms and the assessment of drugs for the treatment of castration resistant prostate cancer.

METHODSWe generated two novel cell line models (LAPC4-CR and VCaP-CR) which were derived by passaging LAPC4 and VCaP cells in vivo and in vitro under castrate conditions. We performed detailed transcriptomic (RNA-seq) and proteomic analyses (SWATH-MS) to delineate expression differences between castration-sensitive and castration-resistant cell lines. Furthermore, we characterized the in vivo and in vitro growth characteristics of the novel cell line models.

RESULTSThe two cell line derivatives LAPC4-CR and VCaP-CR showed castration resistant growth in vitro and in vivo which was only minimally inhibited by AR antagonists, enzalutamide and bicalutamide. High-dose androgen treatment resulted in significant growth arrest of VCaP-CR but not in LAPC4-CR cells. Both cell lines maintained AR expression, but exhibited distinct expression changes on the mRNA and protein level. Integrated analyses including data from LNCaP and the previously described castration resistant LNCaP-abl cells revealed an expression signature of castration resistance.

CONCLUSIONSThe two novel cell line models LAPC4-CR and VCaP-CR and their comprehensive characterization on the RNA and protein level represent important resources to study the molecular mechanisms of castration resistance.
]]></description>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Bhamidipati, A.</dc:creator>
<dc:creator>Tsai, H. K.</dc:creator>
<dc:creator>Esopi, D. M.</dc:creator>
<dc:creator>Vaghasia, A. M.</dc:creator>
<dc:creator>Low, J.-Y.</dc:creator>
<dc:creator>Patel, R. A.</dc:creator>
<dc:creator>Guner, G.</dc:creator>
<dc:creator>Pham, M.-T.</dc:creator>
<dc:creator>Castagna, N.</dc:creator>
<dc:creator>Hicks, J.</dc:creator>
<dc:creator>Wyhs, N.</dc:creator>
<dc:creator>Aebersold, R.</dc:creator>
<dc:creator>De Marzo, A. M.</dc:creator>
<dc:creator>Nelson, W. G.</dc:creator>
<dc:creator>Guo, T.</dc:creator>
<dc:creator>Yegnasubramanian, S.</dc:creator>
<dc:date>2021-07-05</dc:date>
<dc:identifier>doi:10.1101/2021.07.04.450352</dc:identifier>
<dc:title><![CDATA[PHENOTYPIC CHARACTERIZATION OF TWO NOVEL CELL LINE MODELS OF CASTRATION RESISTANT PROSTATE CANCER.]]></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/10.1101/2021.07.07.451375v1?rss=1">
<title>
<![CDATA[
Multivalent designed proteins protect against SARS-CoV-2 variants of concern 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.07.451375v1?rss=1</link>
<description><![CDATA[
Escape variants of SARS-CoV-2 are threatening to prolong the COVID-19 pandemic. To address this challenge, we developed multivalent protein-based minibinders as potential prophylactic and therapeutic agents. Homotrimers of single minibinders and fusions of three distinct minibinders were designed to geometrically match the SARS-CoV-2 spike (S) trimer architecture and were optimized by cell-free expression and found to exhibit virtually no measurable dissociation upon binding. Cryo-electron microscopy (cryoEM) showed that these trivalent minibinders engage all three receptor binding domains on a single S trimer. The top candidates neutralize SARS-CoV-2 variants of concern with IC50 values in the low pM range, resist viral escape, and provide protection in highly vulnerable human ACE2-expressing transgenic mice, both prophylactically and therapeutically. Our integrated workflow promises to accelerate the design of mutationally resilient therapeutics for pandemic preparedness.

One-Sentence SummaryWe designed, developed, and characterized potent, trivalent miniprotein binders that provide prophylactic and therapeutic protection against emerging SARS-CoV-2 variants of concern.
]]></description>
<dc:creator>Hunt, A. C.</dc:creator>
<dc:creator>Case, J. B.</dc:creator>
<dc:creator>Park, Y.-J.</dc:creator>
<dc:creator>Cao, L.</dc:creator>
<dc:creator>Wu, K.</dc:creator>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Bowen, J. E.</dc:creator>
<dc:creator>Yeh, H.-W.</dc:creator>
<dc:creator>Saini, S.</dc:creator>
<dc:creator>Helms, L.</dc:creator>
<dc:creator>Zhao, Y. T.</dc:creator>
<dc:creator>Hsiang, T.-Y.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Goreshnik, I.</dc:creator>
<dc:creator>Kozodoy, L.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Ravichandran, R.</dc:creator>
<dc:creator>Green, L. B.</dc:creator>
<dc:creator>Matochko, W. L.</dc:creator>
<dc:creator>Thomson, C. A.</dc:creator>
<dc:creator>Vogeli, B.</dc:creator>
<dc:creator>Kruger-Gericke, A.</dc:creator>
<dc:creator>VanBlargan, L. A.</dc:creator>
<dc:creator>Chen, R. E.</dc:creator>
<dc:creator>Ying, B.</dc:creator>
<dc:creator>Bailey, A. L.</dc:creator>
<dc:creator>Kafai, N. M.</dc:creator>
<dc:creator>Boyken, S.</dc:creator>
<dc:creator>Ljubetic, A.</dc:creator>
<dc:creator>Edman, N.</dc:creator>
<dc:creator>Ueda, G.</dc:creator>
<dc:creator>Chow, C.</dc:creator>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Panpradist, N.</dc:creator>
<dc:creator>Gale, M.</dc:creator>
<dc:creator>Freedman, B.</dc:creator>
<dc:creator>Lutz, B.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Ruohola-Baker, H.</dc:creator>
<dc:creator>Whelan, S. P. J.</dc:creator>
<dc:creator>Stewart, L.</dc:creator>
<dc:creator>Diamond, M. S.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Jewett, M.</dc:creator>
<dc:date>2021-07-07</dc:date>
<dc:identifier>doi:10.1101/2021.07.07.451375</dc:identifier>
<dc:title><![CDATA[Multivalent designed proteins protect against SARS-CoV-2 variants of concern]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-07</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/10.1101/2021.07.11.451980v1?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/10.1101/2021.07.13.452118v1?rss=1">
<title>
<![CDATA[
Active enhancers strengthen TAD insulation by bRNA mediated CTCF enrichment at the TAD boundaries 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.13.452118v1?rss=1</link>
<description><![CDATA[
Vertebrate genomes are partitioned into Topologically Associating Domains (TADs), which are typically bound by head-to-head pairs of CTCF binding sites. Transcription at TAD boundaries correlates with better insulation, however, it is not known whether the boundary transcripts themselves contribute to boundary function. Here we characterize boundary-associated RNAs genome-wide, focusing on the disease-relevant INK4a/ARF TAD. Using CTCF site deletions and boundary-associated RNA knockdowns, we observe that boundary-associated RNAs facilitate recruitment and clustering of CTCF at TAD borders. The resulting CTCF enrichment enhances TAD insulation, enhancer:promoter interactions and TAD gene expression. Importantly, knockdown of boundary-associated RNAs results in loss of boundary insulation function. Using enhancer deletions and CRISPRi of promoters we show that active TAD enhancers but not promoters induce boundary-associated RNA transcription, thus defining a novel class of regulatory enhancer RNAs.
]]></description>
<dc:creator>Islam, Z.</dc:creator>
<dc:creator>Saravanan, B.</dc:creator>
<dc:creator>Walavalkar, K.</dc:creator>
<dc:creator>Farooq, U.</dc:creator>
<dc:creator>Singh, A. K.</dc:creator>
<dc:creator>Thakur, J.</dc:creator>
<dc:creator>Pandit, A.</dc:creator>
<dc:creator>Sabarinathan, R.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Notani, D.</dc:creator>
<dc:date>2021-07-14</dc:date>
<dc:identifier>doi:10.1101/2021.07.13.452118</dc:identifier>
<dc:title><![CDATA[Active enhancers strengthen TAD insulation by bRNA mediated CTCF enrichment at the TAD boundaries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.13.452235v1?rss=1">
<title>
<![CDATA[
Divergence in dimerization and activity of primate APOBEC3C 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.13.452235v1?rss=1</link>
<description><![CDATA[
The APOBEC3 (A3) family of single-stranded DNA cytidine deaminases are host restriction factors that inhibit lentiviruses, such as HIV-1, in the absence of the Vif protein that causes their degradation. Deamination of cytidine in HIV-1 (-)DNA forms uracil that causes inactivating mutations when uracil is used as a template for (+)DNA synthesis. For APOBEC3C (A3C), the chimpanzee and gorilla orthologues are more active than human A3C, and the Old World Monkey A3C from rhesus macaque (rh) is not active against HIV-1. Multiple integrated analyses determined why rhA3C was not active against HIV-1 and how to increase this activity. Biochemical, virological, and coevolutionary analyses combined with molecular dynamics simulations showed that the key amino acids needed to promote rhA3C antiviral activity also promoted dimerization. Although rhA3C shares a similar dimer interface with hominid A3C, the key amino acid contacts were different. Overall, our results determine the basis for why rhA3C is less active than human A3C, establish the amino acid network for dimerization and increased activity, and track the loss and gain of A3C antiviral activity in primates. The coevolutionary analysis of the A3C dimerization interface provides a basis from which to analyze dimerization interfaces of other A3 family members.
]]></description>
<dc:creator>Gaba, A.</dc:creator>
<dc:creator>Hix, M. A.</dc:creator>
<dc:creator>Suhail, S.</dc:creator>
<dc:creator>Flath, B.</dc:creator>
<dc:creator>Boysan, B.</dc:creator>
<dc:creator>Williams, D. R.</dc:creator>
<dc:creator>Pelletier, T.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Morcos, F.</dc:creator>
<dc:creator>Cisneros, G. A.</dc:creator>
<dc:creator>Chelico, L.</dc:creator>
<dc:date>2021-07-13</dc:date>
<dc:identifier>doi:10.1101/2021.07.13.452235</dc:identifier>
<dc:title><![CDATA[Divergence in dimerization and activity of primate APOBEC3C]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.17.452804v1?rss=1">
<title>
<![CDATA[
ACE2 binding is an ancestral and evolvable trait of sarbecoviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.17.452804v1?rss=1</link>
<description><![CDATA[
Two different sarbecoviruses have caused major human outbreaks in the last two decades1,2. Both these sarbecoviruses, SARS-CoV-1 and SARS-CoV-2, engage ACE2 via the spike receptor-binding domain (RBD)2-6. However, binding to ACE2 orthologs from humans, bats, and other species has been observed only sporadically among the broader diversity of bat sarbecoviruses7-11. Here, we use high-throughput assays12 to trace the evolutionary history of ACE2 binding across a diverse range of sarbecoviruses and ACE2 orthologs. We find that ACE2 binding is an ancestral trait of sarbecovirus RBDs that has subsequently been lost in some clades. Furthermore, we demonstrate for the first time that bat sarbecoviruses from outside Asia can bind ACE2. In addition, ACE2 binding is highly evolvable: for many sarbecovirus RBDs there are single amino-acid mutations that enable binding to new ACE2 orthologs. However, the effects of individual mutations can differ markedly between viruses, as illustrated by the N501Y mutation which enhances human ACE2 binding affinity within several SARS-CoV-2 variants of concern12 but severely dampens it for SARS-CoV-1. Our results point to the deep ancestral origin and evolutionary plasticity of ACE2 binding, broadening consideration of the range of sarbecoviruses with spillover potential.
]]></description>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Zepeda, S. K.</dc:creator>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-07-19</dc:date>
<dc:identifier>doi:10.1101/2021.07.17.452804</dc:identifier>
<dc:title><![CDATA[ACE2 binding is an ancestral and evolvable trait of sarbecoviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.19.452625v1?rss=1">
<title>
<![CDATA[
Vaccinia Virus Arrests and Shifts the Cell Cycle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.19.452625v1?rss=1</link>
<description><![CDATA[
Modulation of the host cell cycle is a common strategy used by viruses to create a pro-replicative environment. To facilitate viral genome replication, vaccinia virus (VACV) has been reported to alter cell cycle regulation and trigger the host cell DNA damage response. However, the cellular factors and viral effectors that mediate these changes remain unknown. Here, we set out to investigate the effect of VACV infection on cell proliferation and host cell cycle progression. Using a subset of VACV mutants we characterize the stage of infection required for inhibition of cell proliferation and define the viral effectors required to dysregulate the host cell cycle. Consistent with previous studies, we show that VACV inhibits, and subsequently shifts the host cell cycle. We demonstrate that these two phenomena are independent of one another, with viral early genes being responsible for cell cycle inhibition, and post-replicative viral gene(s) responsible for the cell cycle shift. Extending previous findings, we show that the viral kinase F10 is required to activate the DNA damage checkpoint and that the viral B1/B12 (pseudo) kinases mediate degradation of checkpoint effectors p53 and p21 during infection. We conclude that VACV modulates host cell proliferation and host cell cycle progression through temporal expression of multiple VACV effector proteins.
]]></description>
<dc:creator>Martin, C. K.</dc:creator>
<dc:creator>Samolej, J.</dc:creator>
<dc:creator>Olson, A. T.</dc:creator>
<dc:creator>Bertoli, C.</dc:creator>
<dc:creator>Wiebe, M. S.</dc:creator>
<dc:creator>de Bruin, R. A. M.</dc:creator>
<dc:creator>Mercer, J.</dc:creator>
<dc:date>2021-07-19</dc:date>
<dc:identifier>doi:10.1101/2021.07.19.452625</dc:identifier>
<dc:title><![CDATA[Vaccinia Virus Arrests and Shifts the Cell Cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.28.454252v1?rss=1">
<title>
<![CDATA[
Systems analysis of immune responses to attenuated P. falciparum malaria sporozoite vaccination reveals excessive inflammatory signatures correlating with impaired immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.28.454252v1?rss=1</link>
<description><![CDATA[
Immunization with radiation-attenuated sporozoites (RAS) can confer sterilizing protection against malaria, although the mechanisms behind this protection are incompletely understood. We performed a systems biology analysis of samples from the Immunization by Mosquito with Radiation Attenuated Sporozoites IMRAS) trial, which comprised P. falciparum RAS-immunized (PfRAS), malaria-naive participants whose protection from malaria infection was subsequently assessed by controlled human malaria infection (CHMI). Blood samples collected after initial PfRAS immunization were analyzed to compare immune responses between protected and non-protected volunteers leveraging integrative analysis of whole blood RNA-seq, high parameter flow cytometry, and single cell CITEseq of PBMCs. This analysis revealed differences in early innate immune responses indicating divergent paths associated with protection. In particular, elevated levels of inflammatory responses early after the initial immunization were detrimental for the development of protective adaptive immunity. Specifically, non-classical monocytes and early type I interferon responses induced within 1 day of PfRAS vaccination correlated with impaired immunity. Non-protected individuals also showed an increase in Th2 polarized T cell responses whereas we observed a trend towards increased Th1 and T-bet+ CD8 T cell responses in protected individuals. Temporal differences in genes associated with natural killer cells suggest an important role in immune regulation by these cells. These findings give insight into the immune responses that confer protection against malaria and may guide further malaria vaccine development.
]]></description>
<dc:creator>Du, Y.</dc:creator>
<dc:creator>Hertoghs, N.</dc:creator>
<dc:creator>Carnes, J.</dc:creator>
<dc:creator>McDermott, S. M.</dc:creator>
<dc:creator>Neal, M. L.</dc:creator>
<dc:creator>Schwedhelm, K. V.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>De Rosa, S. C.</dc:creator>
<dc:creator>Aitchison, J. D.</dc:creator>
<dc:creator>Stuart, K. D.</dc:creator>
<dc:date>2021-07-29</dc:date>
<dc:identifier>doi:10.1101/2021.07.28.454252</dc:identifier>
<dc:title><![CDATA[Systems analysis of immune responses to attenuated P. falciparum malaria sporozoite vaccination reveals excessive inflammatory signatures correlating with impaired immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.30.454539v1?rss=1">
<title>
<![CDATA[
Engineering adoptive T cell therapy to co-opt Fas ligand-mediated death signaling in ovarian cancer enhances therapeutic efficacy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.30.454539v1?rss=1</link>
<description><![CDATA[
BackgroundIn the U.S., more than 50% of ovarian cancer patients die within 5 years of diagnosis, highlighting the need for innovations such as engineered T cell therapies. Mesothelin (Msln) is an attractive immunotherapy target for this cancer, as it is overexpressed by the tumor and contributes to malignant and invasive phenotypes, making antigen loss disadvantageous to the tumor. We previously showed that adoptively transferred T cells engineered to be Msln-specific (TCR1045) preferentially accumulate within established ovarian tumors, delay tumor growth and significantly prolong survival in the ID8VEGF mouse model. However, T cell persistence and anti-tumor activity were not sustained, and we and others have previously detected FasL in the tumor vasculature and the tumor microenvironment (TME) of human and murine ovarian cancers, which can induce apoptosis in infiltrating lymphocytes expressing Fas receptor (Fas).

MethodsTo concurrently overcome this mechanism for potential immune evasion and enhance T cell responses, we generated an immunomodulatory fusion protein (IFP) containing the Fas extracellular binding domain fused to a 4-1BB co-stimulatory domain, rather than the natural death domain. T cells engineered to express TCR1045 alone or in combination with the IFP were transferred into ID8VEGF-tumor bearing mice and evaluated for persistence, proliferation, anti-tumor cytokine production, and therapeutic efficacy.

ResultsRelative to T cells modified only to express TCR1045, T cells engineered to express both TCR1045 and a Fas IFP preferentially persisted in the TME of tumor-bearing mice due to improved T cell proliferation and survival. Moreover, adoptive immunotherapy with IFP+ T cells significantly prolonged survival in tumor-bearing mice, relative to TCR1045 T cells lacking the IFP.

ConclusionsFas/FasL signaling can mediate T cell death in the ovarian cancer microenvironment, as well as induce activation-induced cell death, an apoptotic mechanism responsible for regulating T cell expansion. Upregulation of FasL by tumor cells and tumor vasculature represents a mechanism for protecting growing tumors from attack by tumor-infiltrating lymphocytes. As many solid tumors overexpress FasL, an IFP that converts the Fas-mediated death signal into pro-survival and proliferative signals may provide an opportunity to enhance engineered adoptive T cell therapy against many malignancies.
]]></description>
<dc:creator>Anderson, K. G.</dc:creator>
<dc:creator>Oda, S. K.</dc:creator>
<dc:creator>Bates, B. M.</dc:creator>
<dc:creator>Burnett, M. G.</dc:creator>
<dc:creator>Rodgers Suarez, M.</dc:creator>
<dc:creator>Ruskin, S. L.</dc:creator>
<dc:creator>Greenberg, P. D.</dc:creator>
<dc:date>2021-07-31</dc:date>
<dc:identifier>doi:10.1101/2021.07.30.454539</dc:identifier>
<dc:title><![CDATA[Engineering adoptive T cell therapy to co-opt Fas ligand-mediated death signaling in ovarian cancer enhances therapeutic efficacy]]></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/10.1101/2021.08.02.454829v1?rss=1">
<title>
<![CDATA[
Structural definition of a pan-sarbecovirus neutralizing epitope on the spike S2 subunit. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.02.454829v1?rss=1</link>
<description><![CDATA[
Three highly pathogenic betacoronaviruses have crossed the species barrier and established human-to-human transmission causing significant morbidity and mortality in the past 20 years. The most current and widespread of these is SARS-CoV-2. The identification of CoVs with zoonotic potential in animal reservoirs suggests that additional outbreaks are likely to occur. Evidence suggests that neutralizing antibodies are important for protection against infection with CoVs. Monoclonal antibodies targeting conserved neutralizing epitopes on diverse CoVs can form the basis for prophylaxis and therapeutic treatments and enable the design of vaccines aimed at providing pan-coronavirus protection. To this end, we previously identified a neutralizing monoclonal antibody, CV3-25 that binds to the SARS-CoV-2 fusion machinery, neutralizes the SARS-CoV-2 Beta variant comparably to the ancestral Wuhan Hu-1 strain, cross neutralizes SARS-CoV-1 and displays cross reactive binding to recombinant proteins derived from the spike-ectodomains of HCoV-OC43 and HCoV-HKU1. Here, we show that the neutralizing activity of CV3-25 is also maintained against the Alpha, Delta and Gamma variants of concern as well as a SARS-CoV-like bat coronavirus with zoonotic potential by binding to a conserved linear peptide in the stem-helix region on sarbecovirus spikes. A 1.74[A] crystal structure of a CV3-25/peptide complex demonstrates that CV3-25 binds to the base of the stem helix at the HR2 boundary to an epitope that is distinct from other stem-helix directed neutralizing mAbs. Thus, CV3-25 defines a novel site of sarbecovirus vulnerability that will inform pan-CoV vaccine development.
]]></description>
<dc:creator>Hurlburt, N.</dc:creator>
<dc:creator>Homad, L.</dc:creator>
<dc:creator>Siha, I.</dc:creator>
<dc:creator>Jenewein, M. F.</dc:creator>
<dc:creator>MacCamy, A.</dc:creator>
<dc:creator>Wan, Y.-H.</dc:creator>
<dc:creator>Boonyaratanakornkit, J.</dc:creator>
<dc:creator>Sholukh, A. M.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Burton, D.</dc:creator>
<dc:creator>Andrabi, R.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>McGuire, A.</dc:creator>
<dc:date>2021-08-03</dc:date>
<dc:identifier>doi:10.1101/2021.08.02.454829</dc:identifier>
<dc:title><![CDATA[Structural definition of a pan-sarbecovirus neutralizing epitope on the spike S2 subunit.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.03.454902v1?rss=1">
<title>
<![CDATA[
A distinct core regulatory module enforces oncogene expression in KMT2A-rearranged leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.03.454902v1?rss=1</link>
<description><![CDATA[
A small set of lineage-restricted transcription factors (TFs), termed core regulatory circuitry (CRC), control cell identity and malignant transformation. Here, we integrated gene dependency, chromatin architecture and TF perturbation datasets to characterize 31 core TFs in acute myeloid leukemia (AML). Contrary to a widely accepted model, we detected a modular CRC structure with hierarchically organized, partially redundant and only sparsely interconnected modules of core TFs controlling distinct genetic programs. Rapid TF degradation followed by measurement of genome-wide transcription rates revealed that core TFs directly regulate dramatically fewer genes than previously assumed. Leukemias carrying KMT2A (MLL) rearrangements depend on the IRF8/MEF2 axis to directly enforce expression of the key oncogenes MYC, HOXA9 and BCL2. Our datasets provide an evolving model of CRC organization in human cells, and a resource for further inquiries into and therapeutic targeting of aberrant transcriptional circuits in cancer.
]]></description>
<dc:creator>Harada, T.</dc:creator>
<dc:creator>Heshmati, Y.</dc:creator>
<dc:creator>Kalfon, J.</dc:creator>
<dc:creator>Ferrucio, J. X.</dc:creator>
<dc:creator>Perez, M.</dc:creator>
<dc:creator>Ewers, J.</dc:creator>
<dc:creator>Kossenkov, A.</dc:creator>
<dc:creator>Ellegast, J. M.</dc:creator>
<dc:creator>Yi, J. S.</dc:creator>
<dc:creator>Bowker, A.</dc:creator>
<dc:creator>Zhu, Q.</dc:creator>
<dc:creator>Eagle, K.</dc:creator>
<dc:creator>Dempster, J. M.</dc:creator>
<dc:creator>Kugener, G.</dc:creator>
<dc:creator>Wickramasinghe, J.</dc:creator>
<dc:creator>Herbert, Z. T.</dc:creator>
<dc:creator>Li, C. H.</dc:creator>
<dc:creator>Koren, J. V.</dc:creator>
<dc:creator>Weinstock, D. M.</dc:creator>
<dc:creator>Paralkar, V. R.</dc:creator>
<dc:creator>Nabet, B.</dc:creator>
<dc:creator>Lin, C. Y.</dc:creator>
<dc:creator>Dharia, N. V.</dc:creator>
<dc:creator>Stegmaier, K.</dc:creator>
<dc:creator>Orkin, S. H.</dc:creator>
<dc:creator>Pimkin, M.</dc:creator>
<dc:date>2021-08-04</dc:date>
<dc:identifier>doi:10.1101/2021.08.03.454902</dc:identifier>
<dc:title><![CDATA[A distinct core regulatory module enforces oncogene expression in KMT2A-rearranged leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.05.455215v1?rss=1">
<title>
<![CDATA[
Coordinated Cross-Talk Between the Myc and Mlx Networks in Liver Regeneration and Neoplasia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.05.455215v1?rss=1</link>
<description><![CDATA[
Background & AimsThe c-Myc (Myc) bHLH-ZIP transcription factor is deregulated in most cancers. In association with Max, Myc controls target genes that supervise metabolism, ribosome biogenesis, translation and proliferation. This "Myc Network" cross-talks with the "Mlx Network", which consists of the Myc-like proteins MondoA and ChREBP and Max-like Mlx. Together, this "Extended Myc Network" regulates both common and distinct genes targets. Here we studied the consequence of Myc and/or Mlx ablation in the liver, particularly those pertaining to hepatocyte proliferation, metabolism and spontaneous tumorigenesis.

MethodsWe examined the ability of hepatocytes lacking Mlx (MlxKO) or Myc+Mlx (double KO or DKO) to repopulate the liver over an extended period of time in a murine model of Type I tyrosinemia. We also compared this and other relevant behaviors, phenotypes and transcriptomes of the livers to those from previously characterized MycKO, ChrebpKO and MycKO x ChrebpKO mice.

ResultsHepatocyte regenerative potential deteriorated as the Extended Myc Network was progressively dismantled. Genes and pathways dysregulated in MlxKO and DKO hepatocytes included those pertaining to translation, mitochondrial function and non-alcoholic fatty liver disease (NAFLD). The Myc and Mlx Networks were shown to cross-talk, with the latter playing a disproportionate role in target gene regulation. All cohorts also developed NAFLD and molecular evidence of early steatohepatitis. Finally, MlxKO and DKO mice displayed extensive hepatic adenomatosis.

ConclusionsIn addition to demonstrating cooperation between the Myc and Mlx Networks, this study revealed the latter to be more important in maintaining proliferative, metabolic and translational homeostasis, while concurrently serving as a suppressor of benign tumorigenesis.

SynopsisThe Myc and Mlx Networks exhibit extensive cross-talk and regulate distinct but overlapping sets of transcriptional targets. The current work demonstrates the cooperation between these two Networks in supporting the regenerative capabilities of normal hepatocytes while also revealing that the Mlx Network serves as a suppressor of spontaneous hepatic adenomatosis
]]></description>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>Alencastro, F.</dc:creator>
<dc:creator>Roberts, A.</dc:creator>
<dc:creator>Fiedor, J.</dc:creator>
<dc:creator>Carroll, P.</dc:creator>
<dc:creator>Eisenman, R.</dc:creator>
<dc:creator>Ranganathan, S.</dc:creator>
<dc:creator>Torbenson, M.</dc:creator>
<dc:creator>Duncan, A. W.</dc:creator>
<dc:creator>Prochownik, E. V.</dc:creator>
<dc:date>2021-08-06</dc:date>
<dc:identifier>doi:10.1101/2021.08.05.455215</dc:identifier>
<dc:title><![CDATA[Coordinated Cross-Talk Between the Myc and Mlx Networks in Liver Regeneration and Neoplasia]]></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/10.1101/2021.08.09.453882v1?rss=1">
<title>
<![CDATA[
Quantitative Restoration of Immune Defense in Old Animals Determined by Naive Antigen-Specific CD8 T cell Numbers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.09.453882v1?rss=1</link>
<description><![CDATA[
Older humans and animals often exhibit reduced immune responses to infection and vaccination, and this often directly correlates to the numbers and frequency of naive T (Tn) cells. We found such a correlation between reduced numbers of blood CD8+ Tn cells and severe clinical outcomes of West Nile virus (WNV) in both humans naturally exposed to, and mice experimentally infected with, WNV.

To examine possible causality, we sought to increase the number of CD8 Tn cells by treating C57BL/6 mice with IL-7 complexes (IL-7C, anti-IL-7 mAb bound to IL-7), shown previously to efficiently increase peripheral T cell numbers by homeostatic proliferation. T cells underwent robust expansion following IL-7C administration to old mice increasing the number of total T cells (>four-fold) and NS4b:H-2Db-restricted antigen-specific CD8 T cells (two-fold). This improved the numbers of NS4b-specific CD8 T cells detected at the peak of the response against WNV, but not survival of WNV challenge. IL-7C treated old animals also showed no improvement in WNV-specific effector immunity (neutralizing antibody and in vivo T cell cytotoxicity). To test quantitative limits to which CD8 Tn cell restoration could improve protective immunity, we transferred graded doses of Ag-specific precursors into old mice and showed that injection of 5,400 (but not of 1,800 or 600) adult naive WNV-specific CD8 T cells significantly increased survival after WNV. These results set quantitative limits to the level of Tn reconstitution necessary to improve immune defense in older organisms and are discussed in light of targets of immune reconstitution.
]]></description>
<dc:creator>Uhrlaub, J.</dc:creator>
<dc:creator>Jergovic, M.</dc:creator>
<dc:creator>Bradshaw, C. M.</dc:creator>
<dc:creator>Sonar, S.</dc:creator>
<dc:creator>Coplen, C. P.</dc:creator>
<dc:creator>Dudakov, J.</dc:creator>
<dc:creator>Murray, K. O.</dc:creator>
<dc:creator>Lanteri, M. C.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>van den Brink, M. R. M.</dc:creator>
<dc:creator>Nikolich-Zugich, J.</dc:creator>
<dc:date>2021-08-10</dc:date>
<dc:identifier>doi:10.1101/2021.08.09.453882</dc:identifier>
<dc:title><![CDATA[Quantitative Restoration of Immune Defense in Old Animals Determined by Naive Antigen-Specific CD8 T cell Numbers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.11.455919v1?rss=1">
<title>
<![CDATA[
The light chain of the L9 antibody is critical for binding circumsporozoite protein minor repeats and preventing malaria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.11.455919v1?rss=1</link>
<description><![CDATA[
L9 is a potent human monoclonal antibody (mAb) that preferentially binds two adjacent NVDP minor repeats and cross-reacts with NANP major repeats of the Plasmodium falciparum circumsporozoite protein (PfCSP) on malaria-infective sporozoites. Understanding this mAbs ontogeny and mechanisms of binding PfCSP to neutralize sporozoites will facilitate vaccine development. Here, we isolated mAbs clonally related to L9 and showed that this B-cell lineage has baseline NVDP affinity and evolves to acquire NANP reactivity. Pairing the L9 kappa light chain (L9{kappa}) with clonally-related heavy chains resulted in chimeric mAbs that cross-linked two NVDP, cross-reacted with NANP, and more potently neutralized sporozoites compared to their original light chain. Structural analyses revealed that chimeric mAbs bound the minor repeat motif in a type-1 {beta}-turn seen in other repeat-specific antibodies. These data highlight the importance of L9{kappa} in binding NVDP on PfCSP to neutralize SPZ and suggest that PfCSP-based immunogens might be improved by presenting [&ge;]2 NVDP.
]]></description>
<dc:creator>Wang, L. T.</dc:creator>
<dc:creator>Hurlburt, N. K.</dc:creator>
<dc:creator>Schon, A.</dc:creator>
<dc:creator>Flynn, B. J.</dc:creator>
<dc:creator>Pereira, L. S.</dc:creator>
<dc:creator>Dillon, M.</dc:creator>
<dc:creator>Flores-Garcia, Y.</dc:creator>
<dc:creator>Bonilla, B.</dc:creator>
<dc:creator>Zavala, F.</dc:creator>
<dc:creator>Idris, A. H.</dc:creator>
<dc:creator>Francica, J. R.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>Seder, R. A.</dc:creator>
<dc:date>2021-08-11</dc:date>
<dc:identifier>doi:10.1101/2021.08.11.455919</dc:identifier>
<dc:title><![CDATA[The light chain of the L9 antibody is critical for binding circumsporozoite protein minor repeats and preventing malaria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.12.456166v1?rss=1">
<title>
<![CDATA[
Autoimmune Alleles at the Major Histocompatibility Locus Modify Melanoma Susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.12.456166v1?rss=1</link>
<description><![CDATA[
Autoimmunity and cancer represent two different aspects of immune dysfunction. Autoimmunity is characterized by breakdowns in immune self-tolerance, while impaired immune surveillance can allow for tumorigenesis. The class I major histocompatibility complex (MHC-I), which displays derivatives of the cellular peptidome for immune surveillance by CD8+ T cells, serves as a common genetic link between these conditions. As melanoma-specific CD8+ T-cells have been shown to target melanocyte-specific peptide antigens more often than melanoma-specific antigens, we investigated whether vitiligo and psoriasis predisposing MHC-I alleles conferred a melanoma protective effect. In individuals with cutaneous melanoma from both The Cancer Genome Atlas (N = 451) and an independent validation cohort (N = 586), MHC-I autoimmune allele carrier status was significantly associated with a later age of melanoma diagnosis. Furthermore, MHC-I autoimmune allele carriers were significantly associated with decreased risk of developing melanoma in the Million Veterans Program cohort (OR = 0.962, p = 0.024). Existing melanoma polygenic risk scores (PRS) did not predict autoimmune allele carrier status, suggesting these alleles provide new risk-relevant information. Mechanisms of autoimmune protection were neither associated with improved melanoma-driver mutation association nor improved gene-level conserved antigen presentation relative to common alleles (population frequency > 1%). However, autoimmune alleles showed higher affinity relative to common alleles for particular windows of melanocyte conserved antigens suggesting a potential relationship between antigen processing, binding, and cell-surface presentation. Overall, this study presents evidence that MHC-I autoimmune risk alleles modulate melanoma risk unaccounted for by current PRS.
]]></description>
<dc:creator>Talwar, J.</dc:creator>
<dc:creator>Laub, D.</dc:creator>
<dc:creator>Pagadala, M.</dc:creator>
<dc:creator>Castro, A.</dc:creator>
<dc:creator>Lewis, M.</dc:creator>
<dc:creator>Luebeck, G. E.</dc:creator>
<dc:creator>Morris, G. P.</dc:creator>
<dc:creator>Salem, R. M.</dc:creator>
<dc:creator>Thompson, W. K.</dc:creator>
<dc:creator>Curtius, K.</dc:creator>
<dc:creator>Zanetti, M.</dc:creator>
<dc:creator>Carter, H.</dc:creator>
<dc:date>2021-08-13</dc:date>
<dc:identifier>doi:10.1101/2021.08.12.456166</dc:identifier>
<dc:title><![CDATA[Autoimmune Alleles at the Major Histocompatibility Locus Modify Melanoma Susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.16.456461v1?rss=1">
<title>
<![CDATA[
Unique structure and positive selection promote the rapid divergence of Drosophila Y chromosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.16.456461v1?rss=1</link>
<description><![CDATA[
Y chromosomes across diverse species convergently evolve a gene-poor, heterochromatic organization enriched for duplicated genes, LTR retrotransposable elements, and satellite DNA. Sexual antagonism and a loss of recombination play major roles in the degeneration of young Y chromosomes. However, the processes shaping the evolution of mature, already degenerated Y chromosomes are less well-understood. Because Y chromosomes evolve rapidly, comparisons between closely related species are particularly useful. We generated de novo long read assemblies complemented with cytological validation to reveal Y chromosome organization in three closely related species of the Drosophila simulans complex, which diverged only 250,000 years ago and share >98% sequence identity. We find these Y chromosomes are divergent in their organization and repetitive DNA composition and discover new Y-linked gene families whose evolution is driven by both positive selection and gene conversion. These Y chromosomes are also enriched for large deletions, suggesting that the repair of double-strand breaks on Y chromosomes may be biased toward microhomology-mediated end joining over canonical non-homologous end-joining. We propose that this repair mechanism generally contributes to the convergent evolution of Y chromosome organization.
]]></description>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Gregory, L. E.</dc:creator>
<dc:creator>Gordon, K. E.</dc:creator>
<dc:creator>Meiklejohn, C. D.</dc:creator>
<dc:creator>Larracuente, A.</dc:creator>
<dc:date>2021-08-16</dc:date>
<dc:identifier>doi:10.1101/2021.08.16.456461</dc:identifier>
<dc:title><![CDATA[Unique structure and positive selection promote the rapid divergence of Drosophila Y chromosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.18.455699v1?rss=1">
<title>
<![CDATA[
SOX2-phosphorylation toggles a bistable differentiation-switch in squamous cell carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.18.455699v1?rss=1</link>
<description><![CDATA[
The fate choice between stem cell self-renewal and differentiation is regulated by bistable transcriptional networks, which are balanced in homeostasis and imbalanced in tumors. Yet, how stem cells switch from self-renewal to differentiation remains a conundrum. Here, we discover a molecular mechanism that allows stem cell-like tumor propagating cells (TPCs) in squamous cell carcinomas (SCCs) to switch from a mutually exclusive SOX2-PITX1-TP63 self-renewal circuit to a KLF4 driven differentiation program, dependent on the relative occupancy of a novel Klf4-regulatory enhancer cluster (Klf4EC944) by SOX2 or KLF4, respectively. We find SOX2 occupies this site in TPCs to inhibit Klf4 transcription, but upon phosphorylation SOX2 becomes evicted from Klf4EC944, allowing residual KLF4 to occupy this site instead, boost the expression of KLF4 and its downstream targets, and differentiate self-renewing TPCs into post-mitotic SCC cells. This mechanism allows SOX2 to promote self-renewal and tumor formation, while preserving the differentiation potential in SCC cells. Our data suggest that stochastic cell fate decisions depend on the effective concentration of enzymatically regulated transcription factors. The surprising specificity by which SOX2-phosphorylation governs the bistable Klf4EC944 network-switch in SCCs reveals a conceptual framework for the identification of similar switches in other stem cell and cancer types and their potential development into cell type specific differentiation therapies for diseases in which tissue homeostasis has gone awry.
]]></description>
<dc:creator>Hoang-Phou, S.</dc:creator>
<dc:creator>Sastre-Perona, A.</dc:creator>
<dc:creator>Abbruzzese, M.</dc:creator>
<dc:creator>Ying, Z.</dc:creator>
<dc:creator>Siegle, J.</dc:creator>
<dc:creator>Aranda Orgilles, B.</dc:creator>
<dc:creator>Rocha, P. P.</dc:creator>
<dc:creator>Aifantis, I.</dc:creator>
<dc:creator>Skok, J.</dc:creator>
<dc:creator>Beronja, S.</dc:creator>
<dc:creator>Schober, M.</dc:creator>
<dc:date>2021-08-18</dc:date>
<dc:identifier>doi:10.1101/2021.08.18.455699</dc:identifier>
<dc:title><![CDATA[SOX2-phosphorylation toggles a bistable differentiation-switch in squamous cell carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.18.456876v1?rss=1">
<title>
<![CDATA[
Different adjuvanted pediatric HIV envelope vaccines induced distinct plasma antibody responses despite similar B cell receptor repertoires in infant rhesus macaques. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.18.456876v1?rss=1</link>
<description><![CDATA[
Different HIV vaccine regimens elicit distinct plasma antibody responses in both human and nonhuman primate models. Previous studies in human and non-human primate infants showed that adjuvants influenced the quality of plasma antibody responses induced by pediatric HIV envelope vaccine regimens. We recently reported that use of the 3M052-SE adjuvant and longer intervals between vaccinations are associated with higher magnitude of antibody responses in infant rhesus macaques. However, the impact of different adjuvants in HIV vaccine regimens on the developing infant B cell receptor (BCR) repertoire has not been studied. This study evaluated whether pediatric HIV envelope vaccine regimens with different adjuvants induced distinct antigen-specific memory B cell repertoires and whether specific immunoglobulin (Ig) immunogenetic characteristics are associated with higher magnitude of plasma antibody responses in vaccinated infant rhesus macaques. We utilized archived preclinical pediatric HIV vaccine studies PBMCs and tissue samples from 19 infant rhesus macaques immunized either with (i) HIV Env protein with a squalene adjuvant, (ii) MVA-HIV and Env protein coadministered using a 3-week interval, (iii) MVA-HIV prime/ protein boost with an extended 6-week interval between immunizations, or (iv) with HIV Env administered with 3M-052-SE adjuvant. Frequencies of vaccine-elicited HIV Env-specific memory B cells from PBMCs and tissues were similar across vaccination groups (frequency range of 0.06-1.72%). There was no association between vaccine-elicited antigen-specific memory B cell frequencies and plasma antibody titer or avidity. Moreover, the epitope specificity and Ig immunogenetic features of vaccine-elicited monoclonal antibodies did not differ between the different vaccine regimens. These data suggest that pediatric HIV envelope vaccine candidates with different adjuvants that previously induced higher magnitude and quality of plasma antibody responses in infant rhesus macaques were not driven by distinct antigen-specific memory BCR repertoires.
]]></description>
<dc:creator>Fouda, G. G.</dc:creator>
<dc:creator>Berendam, S. J.</dc:creator>
<dc:creator>Morgan-Asiedu, P. K.</dc:creator>
<dc:creator>Mangan, R. J.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Heimsath, H.</dc:creator>
<dc:creator>Luo, K.</dc:creator>
<dc:creator>Curtis, A. D.</dc:creator>
<dc:creator>Eudailey, J. A.</dc:creator>
<dc:creator>Fox, C. B.</dc:creator>
<dc:creator>Tomai, M. A.</dc:creator>
<dc:creator>Phillips, B.</dc:creator>
<dc:creator>Itell, H. L.</dc:creator>
<dc:creator>Kunz, E.</dc:creator>
<dc:creator>Hudgens, M.</dc:creator>
<dc:creator>Cronin, K.</dc:creator>
<dc:creator>Wiehe, K.</dc:creator>
<dc:creator>Alam, S. M.</dc:creator>
<dc:creator>Van Rompay, K. K. A.</dc:creator>
<dc:creator>De Paris, K.</dc:creator>
<dc:creator>Permar, S. R.</dc:creator>
<dc:creator>Moody, M. A.</dc:creator>
<dc:date>2021-08-20</dc:date>
<dc:identifier>doi:10.1101/2021.08.18.456876</dc:identifier>
<dc:title><![CDATA[Different adjuvanted pediatric HIV envelope vaccines induced distinct plasma antibody responses despite similar B cell receptor repertoires in infant rhesus macaques.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.21.457217v1?rss=1">
<title>
<![CDATA[
Characterization of a vaccine-elicited human antibody with sequence homology to VRC01-class antibodies that binds the C1C2 gp120 domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.21.457217v1?rss=1</link>
<description><![CDATA[
Broadly HIV-1 neutralizing VRC01-class antibodies bind the CD4-binding site of the HIV-1 envelope (Env) and contain VH1-2*02-derived heavy chains paired with light chains expressing five amino acid long CDRL3s. Their unmutated forms do not recognize Env or neutralize HIV-1. The lack of elicitation of VRC01-class antibodies in human clinical trials could potentially be due to the absence of activation of the corresponding naive B cells by the vaccine Env immunogens. To address this point directly, we examined Env-specific BCR sequences from participants in the HVTN 100 clinical trial. Of all the sequences analyzed only one displayed sequence homology to VRC01-class antibodies, but the corresponding antibody (FH1) recognized the C1C2 gp120 domain. For FH1 to switch epitope recognition to the CD4-binding site, alterations in both the CDRH3 and CDRL3 were necessary. Our findings support the use of specifically designed immunogens to activate VRC01-class B cells in future human vaccine trials.
]]></description>
<dc:creator>Gray, M. D.</dc:creator>
<dc:creator>Feng, J.</dc:creator>
<dc:creator>Weidle, C. E.</dc:creator>
<dc:creator>Cohen, K. W.</dc:creator>
<dc:creator>Ballweber-Fleming, L.</dc:creator>
<dc:creator>MacCamy, A. J.</dc:creator>
<dc:creator>Huynh, C. N.</dc:creator>
<dc:creator>Trichka, J. J.</dc:creator>
<dc:creator>Montefiori, D.</dc:creator>
<dc:creator>Ferrari, G.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:date>2021-08-23</dc:date>
<dc:identifier>doi:10.1101/2021.08.21.457217</dc:identifier>
<dc:title><![CDATA[Characterization of a vaccine-elicited human antibody with sequence homology to VRC01-class antibodies that binds the C1C2 gp120 domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.22.457286v1?rss=1">
<title>
<![CDATA[
Coordinately regulated interbacterial antagonism defense pathways constitute a bacterial innate immune system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.22.457286v1?rss=1</link>
<description><![CDATA[
Bacterial survival is fraught with antagonism, including that deriving from viruses and competing bacterial cells1-3 4. It is now appreciated that bacteria mount complex antiviral responses; however, whether a coordinated defense against bacterial threats is undertaken is not well understood. Previously we showed that Pseudomonas aeruginosa possess a danger sensing pathway that is a critical fitness determinant during competition against other bacteria5, 6. Here, we conducted genome-wide screens in P. aeruginosa that reveal three conserved and widespread interbacterial antagonism resistance clusters (arc1-3). We find that although arc1-3 are coordinately activated by the Gac/Rsm danger sensing system, they function independently and provide idiosyncratic defense capabilities, distinguishing them from general stress response pathways. Our findings demonstrate that Arc3 family proteins provide specific protection against phospholipase toxins by preventing the accumulation of lysophospholipids in a manner distinct from previously characterized membrane repair systems. These findings liken the response of P. aeruginosa to bacterial threats to that of eukaryotic innate immunity, wherein threat detection leads to the activation of specialized defense systems.
]]></description>
<dc:creator>Ting, S.-Y.</dc:creator>
<dc:creator>LaCourse, K. D.</dc:creator>
<dc:creator>Ledvina, H. E.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Radey, M. C.</dc:creator>
<dc:creator>Kulasekara, H. D.</dc:creator>
<dc:creator>Somavanshi, R.</dc:creator>
<dc:creator>Bertolli, S. K.</dc:creator>
<dc:creator>Gallagher, L. A.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Penewit, K. M.</dc:creator>
<dc:creator>Salipante, S. J.</dc:creator>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Peterson, S. B.</dc:creator>
<dc:creator>Mougous, J. D.</dc:creator>
<dc:date>2021-08-23</dc:date>
<dc:identifier>doi:10.1101/2021.08.22.457286</dc:identifier>
<dc:title><![CDATA[Coordinately regulated interbacterial antagonism defense pathways constitute a bacterial innate immune system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.23.457421v1?rss=1">
<title>
<![CDATA[
HIV-1 Vif gained breadth in APOBEC3G specificity after cross-species transmission of its precursors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.23.457421v1?rss=1</link>
<description><![CDATA[
APOBEC3G (A3G) is a host-encoded cytidine deaminase that potently restricts retroviruses, such as HIV-1, and depends on its ability to package into virions. As a consequence of this, HIV-1 protein Vif has evolved to antagonize human A3G by targeting it for ubiquitination and subsequent degradation. There is an ancient arms-race between Vif and A3G highlighted by amino acids 128 and 130 in A3G that have evolved under positive selection due to Vif-mediated selective pressure in Old World primates. Nonetheless, not all possible amino acid combinations at these sites have been sampled by nature and it is not clear the evolutionary potential of species to resist Vif antagonism. To explore the evolutionary space of positively selected sites in the Vif-binding region of A3G, we designed a combinatorial mutagenesis screen to introduce all 20 amino acids at sites 128 and 130. Our screen uncovered mutants of A3G with several interesting phenotypes, including loss of stability and resistance of Vif antagonism. However, HIV-1 Vif exhibited remarkable flexibility in antagonizing A3G 128 and 130 mutants, which significantly reduces viable Vif resistance strategies for hominid primates. Importantly, we find that broadened Vif specificity was conferred through Loop 5 adaptations that were required for cross-species adaptation from Old World monkey A3G to hominid A3G. Our evidence suggests that Vif adaptation to novel A3G interfaces during cross-species transmission may train Vif towards broadened specificity that can further facilitate cross-species transmissions and raise the barrier to host resistance.
]]></description>
<dc:creator>Chesarino, N. M.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2021-08-23</dc:date>
<dc:identifier>doi:10.1101/2021.08.23.457421</dc:identifier>
<dc:title><![CDATA[HIV-1 Vif gained breadth in APOBEC3G specificity after cross-species transmission of its precursors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.02.458741v1?rss=1">
<title>
<![CDATA[
Activation of the Zinc-sensing receptor GPR39 promotes T cell reconstitution after hematopoietic stem cell transplant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.02.458741v1?rss=1</link>
<description><![CDATA[
Prolonged lymphopenia represents a major clinical problem after cytoreductive therapies such as chemotherapy and the conditioning required for hematopoietic stem cell transplant (HCT), contributing toward the risk of infections and malignant relapse. Restoration of T cell immunity is dependent on tissue regeneration in the thymus, the primary site of T cell development; although the capacity of the thymus to repair itself diminishes over lifespan. However, although boosting thymic function and T cell reconstitution is of considerable clinical importance, there are currently no approved therapies for treating lymphopenia. Here we found that Zinc (Zn), is critically important for both normal T cell development as well as repair after acute damage. Accumulated Zn in thymocytes during development was released into the extracellular milieu after HCT conditioning, where it triggered regeneration by stimulating endothelial cell-production of BMP4 via the cell surface receptor GPR39. Dietary supplementation of Zn was sufficient to promote thymic function in a mouse model of allogeneic HCT, including enhancing the number of recent thymic emigrants in circulation; although direct targeting of GPR39 with a small molecule agonist enhanced thymic function without the need for prior Zn accumulation in thymocytes. Together, these findings not only define an important pathway underlying tissue regeneration, but also offer an innovative preclinical approach to treat lymphopenia in HCT recipients.

KEY POINTSO_LIThymocytes release zinc after HCT conditioning is sensed by GPR39 and promotes epithelial repair
C_LIO_LIPharmacologic stimulation of GPR39 promotes T cell reconstitution after HCT
C_LI
]]></description>
<dc:creator>Iovino, L.</dc:creator>
<dc:creator>Cooper, K.</dc:creator>
<dc:creator>deRoos, P.</dc:creator>
<dc:creator>Kinsella, S.</dc:creator>
<dc:creator>Evandy, C.</dc:creator>
<dc:creator>Ugrai, T.</dc:creator>
<dc:creator>Mazziotta, F.</dc:creator>
<dc:creator>Ensbey, K. S.</dc:creator>
<dc:creator>Granadier, D.</dc:creator>
<dc:creator>Hopwo, K.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Gagnon, A.</dc:creator>
<dc:creator>Galimberti, S.</dc:creator>
<dc:creator>Petrini, M.</dc:creator>
<dc:creator>Hill, G. R.</dc:creator>
<dc:creator>Dudakov, J. A.</dc:creator>
<dc:date>2021-09-03</dc:date>
<dc:identifier>doi:10.1101/2021.09.02.458741</dc:identifier>
<dc:title><![CDATA[Activation of the Zinc-sensing receptor GPR39 promotes T cell reconstitution after hematopoietic stem cell transplant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.02.458791v1?rss=1">
<title>
<![CDATA[
Multiplexed triage of candidate biomarkers in plasma using internal standard triggered-parallel reaction monitoring mass spectrometry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.02.458791v1?rss=1</link>
<description><![CDATA[
Despite advances in proteomic technologies, clinical translation of plasma biomarkers remains low, partly due to a major bottleneck between the discovery of candidate biomarkers and downstream costly clinical validation studies. Due to a dearth of multiplexable assays, generally only a few candidate biomarkers are tested, and the validation success rate is accordingly low. Here, we demonstrate the capability of internal standard triggered-parallel reaction monitoring (IS-PRM) to prioritize candidate biomarkers for validation studies. A 5,176-plex assay coupling immunodepletion and fractionation with IS-PRM was developed and implemented in human plasma to quantify peptides representing 1,314 breast cancer biomarker candidates. Compared to prior approaches using data-dependent analysis, IS-PRM showed improved sensitivity (912 vs 295 proteins quantified) and precision (CV 0.1 vs 0.27) enabling rank-ordering of candidate biomarkers for validation studies. The assay greatly expands capabilities for quantification of large numbers of proteins and is well suited for prioritization of viable candidate biomarkers.
]]></description>
<dc:creator>Kennedy, J. J.</dc:creator>
<dc:creator>Whiteaker, J. R.</dc:creator>
<dc:creator>Ivey, R. G.</dc:creator>
<dc:creator>Burian, A.</dc:creator>
<dc:creator>Chowdhury, S.</dc:creator>
<dc:creator>Tsai, C.-F.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Lin, C.</dc:creator>
<dc:creator>Murillo, O.</dc:creator>
<dc:creator>Lundeen, R. A.</dc:creator>
<dc:creator>Jones, L. A.</dc:creator>
<dc:creator>Gafken, P. R.</dc:creator>
<dc:creator>Longton, G.</dc:creator>
<dc:creator>Rodland, K. D.</dc:creator>
<dc:creator>Skates, S.</dc:creator>
<dc:creator>Landua, J.</dc:creator>
<dc:creator>Wang, P.</dc:creator>
<dc:creator>Lewis, M. T.</dc:creator>
<dc:creator>Paulovich, A. G.</dc:creator>
<dc:date>2021-09-03</dc:date>
<dc:identifier>doi:10.1101/2021.09.02.458791</dc:identifier>
<dc:title><![CDATA[Multiplexed triage of candidate biomarkers in plasma using internal standard triggered-parallel reaction monitoring mass spectrometry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.09.459676v1?rss=1">
<title>
<![CDATA[
Pro-apoptotic caspase deficiency reveals a cell-extrinsic mechanism of NK cell regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.09.459676v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWRegulated cell death is essential for the maintenance of cellular and tissue homeostasis. In the hematopoietic system, genetic defects in apoptotic cell death generally produce the accumulation of immune cells, inflammation and autoimmunity. In contrast, we found that genetic deletion of caspases of the mitochondrial apoptosis pathway reduces natural killer (NK) cell numbers and makes NK cells functionally defective in vivo and in vitro. Caspase deficiency results in constitutive activation of a type I interferon (IFN) response, due to leakage of mitochondrial DNA and activation of the cGAS/STING pathway. The NK cell defect in caspase-deficient mice is independent of the type I IFN response, but the phenotype is partially rescued by cGAS or STING deficiency. Finally, caspase deficiency alters NK cells in a cell-extrinsic manner. Type I IFNs and NK cells are two essential effectors of antiviral immunity, and our results demonstrate that they are both regulated in a caspase-dependent manner. Beyond caspase-deficient animals, our observations may have implications in infections that trigger mitochondrial stress and caspase-dependent cell death.
]]></description>
<dc:creator>Olsen, T. M.</dc:creator>
<dc:creator>Tan, W. H.</dc:creator>
<dc:creator>Knudsen, A. C.</dc:creator>
<dc:creator>Rongvaux, A.</dc:creator>
<dc:date>2021-09-10</dc:date>
<dc:identifier>doi:10.1101/2021.09.09.459676</dc:identifier>
<dc:title><![CDATA[Pro-apoptotic caspase deficiency reveals a cell-extrinsic mechanism of NK cell regulation]]></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/10.1101/2021.09.09.459682v1?rss=1">
<title>
<![CDATA[
A faithful in vivo model of human macrophages in metastatic melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.09.459682v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWDespite recent therapeutic progress, advanced melanoma remains lethal for many patients. The composition of the immune tumor microenvironment (TME) has decisive impacts on therapy response and disease outcome. High dimensional analyses of patient samples can reveal the composition and heterogeneity of the immune TME. In particular, macrophages are known for their cancer-supportive role, but the underlying mechanisms are incompletely understood, and experimental in vivo systems are needed to test the functional properties of these cells. We characterized a humanized mouse model, reconstituted with a human immune system and a human melanoma, in which: (1) human macrophages support metastatic spread of the tumor; and (2) tumor-infiltrating macrophages have a specific transcriptional signature that faithfully represents the transcriptome of macrophages from patient melanoma samples and is associated with shorter survival. This model complements patient sample analyses, enabling the elucidation of fundamental principles in melanoma biology, and the development and evaluation of candidate therapies.
]]></description>
<dc:creator>Voillet, V.</dc:creator>
<dc:creator>Berger, T. R.</dc:creator>
<dc:creator>McKenna, K. M.</dc:creator>
<dc:creator>Paulson, K. G.</dc:creator>
<dc:creator>Smythe, K. S.</dc:creator>
<dc:creator>Hunter, D. S.</dc:creator>
<dc:creator>Valente, W. J.</dc:creator>
<dc:creator>Weaver, S. J.</dc:creator>
<dc:creator>Campbell, J. S.</dc:creator>
<dc:creator>Kim, T. S.</dc:creator>
<dc:creator>Byrd, D. R.</dc:creator>
<dc:creator>Bielas, J. H.</dc:creator>
<dc:creator>Pierce, R. H.</dc:creator>
<dc:creator>Chapuis, A. G.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Rongvaux, A.</dc:creator>
<dc:date>2021-09-11</dc:date>
<dc:identifier>doi:10.1101/2021.09.09.459682</dc:identifier>
<dc:title><![CDATA[A faithful in vivo model of human macrophages in metastatic melanoma]]></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/10.1101/2021.09.11.459844v1?rss=1">
<title>
<![CDATA[
Rapid and parallel adaptive mutations in spike S1 drive clade success in SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.11.459844v1?rss=1</link>
<description><![CDATA[
Given the importance of variant SARS-CoV-2 viruses with altered receptor-binding or antigenic phenotypes, we sought to quantify the degree to which adaptive evolution is driving accumulation of mutations in the SARS-CoV-2 genome. Here we assessed adaptive evolution across genes in the SARS-CoV-2 genome by correlating clade growth with mutation accumulation as well as by comparing rates of nonsynonymous to synonymous divergence, clustering of mutations across the SARS-CoV-2 phylogeny and degree of convergent evolution of individual mutations. We find that spike S1 is the focus of adaptive evolution, but also identify positively-selected mutations in other genes that are sculpting the evolutionary trajectory of SARS-CoV-2. Adaptive changes in S1 accumulated rapidly, resulting in a remarkably high ratio of nonsynonymous to synonymous divergence that is 2.5X greater than that observed in HA1 at the beginning of the 2009 H1N1 pandemic.
]]></description>
<dc:creator>Kistler, K.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:date>2021-09-12</dc:date>
<dc:identifier>doi:10.1101/2021.09.11.459844</dc:identifier>
<dc:title><![CDATA[Rapid and parallel adaptive mutations in spike S1 drive clade success in SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.14.458942v1?rss=1">
<title>
<![CDATA[
Convergent clonal selection of donor- and recipient-derived CMV-specific T cells in hematopoietic stem cell transplant patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.14.458942v1?rss=1</link>
<description><![CDATA[
Competition between antigen-specific T cells for peptide:MHC (p:MHC) complexes shapes the ensuing T cell response. Mouse model studies provided compelling evidence that competition is a highly effective mechanism controlling the activation of naive T cells. However, assessing the effect of T cell competition in the context of a human infection requires defined pathogen kinetics and trackable naive and memory T cell populations of defined specificity. A unique cohort of non-myeloablative hematopoietic stem cell transplant (nmHSCT) patients allowed us to assess T cell competition in response to CMV reactivation, which was documented with detailed virology data. In our cohort, HSCT donors and recipients were CMV-seronegative and -positive, respectively, thus providing genetically distinct memory and naive T cell populations. We used single-cell transcriptomics to track donor versus recipient-derived T cell clones over the course of 90 days. We found that donor-derived T cell clones proliferated and expanded substantially following CMV-reactivation. However, for immunodominant CMV epitopes, recipient-derived memory T cells remained the overall dominant population. This dominance was maintained despite more robust clonal expansion of donor-derived T cells in response to CMV reactivation. Interestingly, the donor-derived T cells that were recruited into these immunodominant memory populations shared strikingly similar TCR properties with the recipient-derived memory T cells. This selective recruitment of identical and nearly identical clones from the naive into the immunodominant memory T cell pool suggests that competition does not interfere with rejuvenating a memory T cell population, but results in selection of convergent clones to the memory T cell pool.

SignificanceAn existing memory T cell population specific for a single epitope is sufficient to effectively curtail responses to any new antigens if the original epitope is present in a vaccination regimen or heterologous infections. We asked if T cell competition precludes recruitment of any new, naive T cells to an existing memory T cell pool in context of CMV-specific T cell responses in a cohort of transplant patients. Our data indicate that competition does not prevent recruitment of naive T cells into the memory T cell pool, but selects for T cells with nearly or fully congruent T cell receptor specificities. We discuss the implications of rejuvenating a memory T cell pool while preserving the T cell receptor repertoire.
]]></description>
<dc:creator>Erickson, J. R.</dc:creator>
<dc:creator>Stevens-Ayers, T.</dc:creator>
<dc:creator>Mair, F.</dc:creator>
<dc:creator>Edmison, B.</dc:creator>
<dc:creator>Boeckh, M.</dc:creator>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:date>2021-09-15</dc:date>
<dc:identifier>doi:10.1101/2021.09.14.458942</dc:identifier>
<dc:title><![CDATA[Convergent clonal selection of donor- and recipient-derived CMV-specific T cells in hematopoietic stem cell transplant patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.15.460536v1?rss=1">
<title>
<![CDATA[
Mechanism of broad-spectrum Cas9 inhibition by AcrIIA11 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.15.460536v1?rss=1</link>
<description><![CDATA[
Mobile genetic elements evade CRISPR-Cas adaptive immunity by encoding anti-CRISPR proteins (Acrs). Acrs inactivate CRISPR-Cas systems via diverse mechanisms but are generally specific for a narrow subset of Cas nucleases that share high sequence similarity. Here, we demonstrate that AcrIIA11 inhibits diverse Cas9 sub-types in vitro and human cells. Single-molecule fluorescence imaging reveals that AcrIIA11 interferes with the first steps of target search by reducing S. aureus Cas9s diffusion on non-specific DNA. DNA cleavage is inhibited because the AcrIIA11:Cas9 complex is kinetically trapped at PAM-rich decoy sites, preventing Cas9 from reaching its target. This work establishes that DNA trapping can be used to inhibit a broad spectrum of Cas9 orthologs in vitro and during mammalian genome editing.
]]></description>
<dc:creator>Dillard, K. E.</dc:creator>
<dc:creator>Terrace, C.</dc:creator>
<dc:creator>Javanmardi, K.</dc:creator>
<dc:creator>Kim, W.</dc:creator>
<dc:creator>Forsberg, K. J.</dc:creator>
<dc:creator>Finkelstein, I. J.</dc:creator>
<dc:date>2021-09-15</dc:date>
<dc:identifier>doi:10.1101/2021.09.15.460536</dc:identifier>
<dc:title><![CDATA[Mechanism of broad-spectrum Cas9 inhibition by AcrIIA11]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.17.460747v1?rss=1">
<title>
<![CDATA[
Combining genotypes and T cell receptor distributions to infer genetic loci determining V(D)J recombination probabilities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.17.460747v1?rss=1</link>
<description><![CDATA[
Every T cell receptor (TCR) repertoire is shaped by a complex probabilistic tangle of genetically determined biases and immune exposures. T cells combine a random V(D)J recombination process with a selection process to generate highly diverse and functional TCRs. The extent to which an individuals genetic background is associated with their resulting TCR repertoire diversity has yet to be fully explored. Using a previously published repertoire sequencing dataset paired with high-resolution genome-wide genotyping from a large human cohort, we infer specific genetic loci associated with V(D)J recombination probabilities using genome-wide association inference. We show that V(D)J gene usage profiles are associated with variation in the TCRB locus and, specifically for the functional TCR repertoire, variation in the major histocompatibility complex locus. Further, we identify specific variations in the genes encoding the Artemis protein and the TdT protein to be associated with biasing junctional nucleotide deletion and N-insertion, respectively. These results refine our understanding of genetically-determined TCR repertoire biases by confirming and extending previous studies on the genetic determinants of V(D)J gene usage and providing the first examples of trans genetic variants which are associated with modifying junctional diversity. Together, these insights lay the groundwork for further explorations into how immune responses vary between individuals.
]]></description>
<dc:creator>Russell, M. L.</dc:creator>
<dc:creator>Souquette, A.</dc:creator>
<dc:creator>Levine, D. M.</dc:creator>
<dc:creator>Allen, E. K.</dc:creator>
<dc:creator>Kuan, G.</dc:creator>
<dc:creator>Simon, N.</dc:creator>
<dc:creator>Balmaseda, A.</dc:creator>
<dc:creator>Gordon, A.</dc:creator>
<dc:creator>Thomas, P.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Bradley, P. G.</dc:creator>
<dc:date>2021-09-20</dc:date>
<dc:identifier>doi:10.1101/2021.09.17.460747</dc:identifier>
<dc:title><![CDATA[Combining genotypes and T cell receptor distributions to infer genetic loci determining V(D)J recombination probabilities]]></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/10.1101/2021.09.18.459450v1?rss=1">
<title>
<![CDATA[
SL-Cloud: A Computational Resource to Support Synthetic Lethal Interaction Discovery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.18.459450v1?rss=1</link>
<description><![CDATA[
Synthetic lethal interactions (SLIs), genetic interactions in which the simultaneous inactivation of two genes leads to a lethal phenotype, are promising targets for therapeutic intervention in cancer, as exemplified by the recent success of PARP inhibitors in treating BRCA1/2-deficient tumors. We present SL-Cloud, an integrated resource and framework to facilitate the prediction of context-specific SLIs by using cloud-based technologies. This resource addresses two main challenges related to SLI inference: the need to wrangle and preprocess large multi-omic datasets and the multiple comparable prediction approaches available. We demonstrate the utility of this resource by using a set of DNA damage repair genes as the basis for predicting potential SLI partners, using multiple computational strategies. Context-specific synthetic lethality potential can also be compared using the framework. We demonstrate various use cases for our cloud-based computational resource and the utility of this approach for customizable and extensible computational inference of SLIs.
]]></description>
<dc:creator>Tercan, B.</dc:creator>
<dc:creator>Qin, G.</dc:creator>
<dc:creator>Kim, T.</dc:creator>
<dc:creator>Aguilar, B.</dc:creator>
<dc:creator>Kemp, C. J.</dc:creator>
<dc:creator>Chambwe, N.</dc:creator>
<dc:creator>Shmulevich, I.</dc:creator>
<dc:date>2021-09-19</dc:date>
<dc:identifier>doi:10.1101/2021.09.18.459450</dc:identifier>
<dc:title><![CDATA[SL-Cloud: A Computational Resource to Support Synthetic Lethal Interaction Discovery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.21.461216v1?rss=1">
<title>
<![CDATA[
Brain I3 Binding Protein regulates K-Ras4B membrane localization and signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.21.461216v1?rss=1</link>
<description><![CDATA[
Membrane localization of Ras proteins is necessary for their biological functions and oncogenic activity. We report here on the identification of Brain I3 Binding Protein (BRI3BP) as a novel binding partner for Ras. We show that K-Ras4B plasma membrane localization and biological function are reduced in the absence of BRI3BP. BRI3BP interacts with K-Ras4B and K-Ras4A and our data suggest that BRI3BP operates within the recycling endosomal compartment to regulate K-Ras localization to the plasma membrane. This study uncovers a new regulatory protein for Ras membrane localization.
]]></description>
<dc:creator>McCabe, I.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Cooper, J. A.</dc:creator>
<dc:creator>Turner, D. L.</dc:creator>
<dc:creator>Vojtek, A. B.</dc:creator>
<dc:date>2021-09-22</dc:date>
<dc:identifier>doi:10.1101/2021.09.21.461216</dc:identifier>
<dc:title><![CDATA[Brain I3 Binding Protein regulates K-Ras4B membrane localization and signaling]]></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/10.1101/2021.09.21.461235v1?rss=1">
<title>
<![CDATA[
Hematopoietic recovery after transplantation is primarily derived from the stochastic contribution of hematopoietic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.21.461235v1?rss=1</link>
<description><![CDATA[
Reconstitution after hematopoietic stem cell (HSC) transplantation is assumed to occur in two distinct phases: initial recovery mediated by short-term progenitors and long-term repopulation by multipotent HSCs which do not contribute to hematopoietic reconstitution during the first 6-9 months. We have previously reported the transplantation and exclusive engraftment of the HSC-enriched CD34+CD45RA-CD90+ phenotype in a nonhuman primate model. Here, we closely followed the clonal diversity and kinetics in these animals. Enhanced sampling and high density clonal tracking within the first 3 month revealed that multipotent HSCs actively contributed to the early phases of neutrophil recovery and became the dominant source for blood cells as early as 50 days after transplant. Longitudinal changes in clonal diversity supported a stochastic engraftment of HSCs with the majority of HSCs clones vanishing early during neutrophil recovery and a smaller fraction of HSC clones expanding into bigger pools to support long-term hematopoiesis. In contrast to the bi-phasic model, we propose that hematopoietic recovery after myeloablation and transplantation is primarily derived from HSCs in a stochastic manner rather than in two phases by independent cell populations.
]]></description>
<dc:creator>Radtke, S.</dc:creator>
<dc:creator>Enstrom, M.</dc:creator>
<dc:creator>Pande, D. P.</dc:creator>
<dc:creator>Cui, M. L.</dc:creator>
<dc:creator>Madhu, R.</dc:creator>
<dc:creator>Perez, A.</dc:creator>
<dc:creator>Kiem, H.-P.</dc:creator>
<dc:date>2021-09-21</dc:date>
<dc:identifier>doi:10.1101/2021.09.21.461235</dc:identifier>
<dc:title><![CDATA[Hematopoietic recovery after transplantation is primarily derived from the stochastic contribution of hematopoietic stem cells]]></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/10.1101/2021.09.23.461592v1?rss=1">
<title>
<![CDATA[
Batch effects removal for microbiome data via conditional quantile regression (ConQuR) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.23.461592v1?rss=1</link>
<description><![CDATA[
Batch effects in microbiome data arise from differential processing of specimens and can lead to spurious findings and obscure true signals. Most existing strategies for mitigating batch effects rely on approaches designed for genomic analysis, failing to address the zero-inflated and over-dispersed microbiome data. Strategies tailored for microbiome data are restricted to association testing, failing to allow other analytic goals such as visualization. We develop the Conditional Quantile Regression (ConQuR) approach to remove microbiome batch effects using a two-part quantile regression model. It is a fundamental advancement in the field because it is the first comprehensive method that accommodates the complex distributions of microbial read counts, and it generates batch-removed zero-inflated read counts that can be used in and benefit all usual subsequent analyses. We apply ConQuR to real microbiome data sets and demonstrate its state-of-the-art performance in removing batch effects while preserving or even amplifying the signals of interest.
]]></description>
<dc:creator>Ling, W.</dc:creator>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Lulla, A.</dc:creator>
<dc:creator>Plantinga, A. M.</dc:creator>
<dc:creator>Fu, W.</dc:creator>
<dc:creator>Zhang, A.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Randolph, T.</dc:creator>
<dc:creator>Koay, W. L. A.</dc:creator>
<dc:creator>White, J. R.</dc:creator>
<dc:creator>Launer, L. J.</dc:creator>
<dc:creator>Fodor, A. A.</dc:creator>
<dc:creator>Meyer, K. A.</dc:creator>
<dc:creator>Wu, M. C.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.23.461592</dc:identifier>
<dc:title><![CDATA[Batch effects removal for microbiome data via conditional quantile regression (ConQuR)]]></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/10.1101/2021.09.24.461596v1?rss=1">
<title>
<![CDATA[
Prioritizing candidate peptides for cancer vaccines by PEPPRMINT: a statistical model to predict peptide presentation by HLA-I proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.24.461596v1?rss=1</link>
<description><![CDATA[
Recent development of cancer immunotherapy has opened unprecedented avenues to eliminate tumor cells using the human immune system. Cancer vaccines composed of neoantigens, or peptides unique to tumor cells due to somatic mutations, have emerged as a promising approach to activate or strengthen the immune response against cancer. A key step to identifying neoantigens is computationally predicting which somatically mutated peptides are presented on the cell surface by a human leukocyte antigen (HLA). Computational prediction relies on large amounts of high-quality training data, such as mass spectrometry data of peptides presented by one of several HLAs in living cells. We developed a complete pipeline to prioritize neoantigens for cancer vaccines. A key step of our pipeline is PEPPRMINT (PEPtide PResentation using a MIxture model and Neural neTwork), a model designed to exploit mass spectrometry data to predict peptide presentation by HLAs. We applied our pipeline to DNA sequencing data of 60 melanoma patients and identified a group of neoantigens that were more immunogenic in tumor cells than in normal cells. Additionally, the neoantigen burden estimated by PEPPRMINT was significantly associated with activity of the immune system, suggesting these neoantigens could induce an immune response.
]]></description>
<dc:creator>Zhou, L. Y.</dc:creator>
<dc:creator>Zou, F.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2021-09-26</dc:date>
<dc:identifier>doi:10.1101/2021.09.24.461596</dc:identifier>
<dc:title><![CDATA[Prioritizing candidate peptides for cancer vaccines by PEPPRMINT: a statistical model to predict peptide presentation by HLA-I proteins]]></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/10.1101/2021.09.24.461735v1?rss=1">
<title>
<![CDATA[
Single-cell RNA Sequencing Reveals Immunosuppressive Myeloid Cell Diversity and Restricted Cytotoxic Effector Cell Trafficking and Activation During Malignant Progression in Glioma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.24.461735v1?rss=1</link>
<description><![CDATA[
Low grade gliomas (LGG) account for about two-thirds of all glioma diagnoses in adolescents and young adults (AYA) and malignant progression of these patients leads to dismal outcomes. Recent studies have shown the importance of the dynamic tumor microenvironment in high-grade gliomas (HGG), yet its role is still poorly understood in low-grade glioma malignant progression. Here, we investigated the heterogeneity of the immune microenvironment using a platelet-derived growth factor (PDGF)-driven RCAS (replication-competent ASLV long terminal repeat with a splice acceptor) glioma model that recapitulates the malignant progression of low to high-grade glioma in humans and also provides a model system to characterize immune cell trafficking and evolution. To illuminate changes in the immune cell landscape during tumor progression, we performed single-cell RNA sequencing on immune cells isolated from animals bearing no tumor (NT), LGG and HGG, with a particular focus on the myeloid cell compartment, which is known to mediate glioma immunosuppression. LGGs demonstrated significantly increased infiltrating T cells, CD4 T cells, CD8 T cells, B cells, and natural killer cells in the tumor microenvironment, whereas HGGs significantly abrogated this infiltration. Our study identified two distinct macrophage clusters in the tumor microenvironment; one cluster appeared to be bone marrow-derived while another was defined by overexpression of Trem2, a marker of tumor associated macrophages. Our data demonstrates that these two distinct macrophage clusters show an immune-activated phenotype (Stat1, Tnf, Cxcl9 and Cxcl10) in LGG which evolves to an immunosuppressive state (Lgals3, Apoc1 and Id2) in HGG that restricts T cell recruitment and activation. We identified CD74 and macrophage migration inhibition factor (MIF) as potential targets for these distinct macrophage populations. Interestingly, these results were mirrored by our analysis of the TCGA dataset, which demonstrated a statistically significant association between CD74 overexpression and decreased overall survival in AYA patients with grade II gliomas. Targeting immunosuppressive myeloid cells and intra-tumoral macrophages within this therapeutic window may ameliorate mechanisms associated with immunosuppression before and during malignant progression.
]]></description>
<dc:creator>Rajendran, S.</dc:creator>
<dc:creator>Peterson, C.</dc:creator>
<dc:creator>Canella, A.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Gross, A.</dc:creator>
<dc:creator>Cam, M.</dc:creator>
<dc:creator>Serin Harmanci, A.</dc:creator>
<dc:creator>Distefano, R.</dc:creator>
<dc:creator>Nigita, G.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Miller, K. E.</dc:creator>
<dc:creator>Elemento, O.</dc:creator>
<dc:creator>Roberts, R. D.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:creator>Rao, G.</dc:creator>
<dc:creator>Mardis, E. R.</dc:creator>
<dc:creator>Rajappa, P.</dc:creator>
<dc:date>2021-09-25</dc:date>
<dc:identifier>doi:10.1101/2021.09.24.461735</dc:identifier>
<dc:title><![CDATA[Single-cell RNA Sequencing Reveals Immunosuppressive Myeloid Cell Diversity and Restricted Cytotoxic Effector Cell Trafficking and Activation During Malignant Progression in Glioma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.26.461847v1?rss=1">
<title>
<![CDATA[
An innate immune activation state prior to vaccination predicts responsiveness to multiple vaccines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.26.461847v1?rss=1</link>
<description><![CDATA[
Many factors determine whether an individual responding to vaccination will generate an immune response that can lead to protection. Several studies have shown that the pre-vaccination immune state is associated with the antibody response to vaccines. However, the generalizability and mechanisms that underlie this association remain poorly defined. Here, we sought to identify a common pre-vaccination signature and mechanisms that could predict the immune response across a wide variety of vaccines. We leveraged the "Immune Signatures Data Resource" created by the NIH Human Immunology Project Consortium (HIPC) to integrate data from 28 studies involving 13 different vaccines and associate the blood transcriptional status of 820 healthy young adults with their responses. An unsupervised analysis of blood transcriptional profiles across studies revealed three distinct pre-vaccination states, characterized by the differential expression of genes associated with a pro-inflammatory response, cell proliferation, and metabolism alterations downstream of NF{kappa}B and IRF7. Innate and adaptive immune cell subset-specific genes were also associated with the three pre-vaccination states. Importantly, individuals whose pre-vaccination state was enriched in pro-inflammatory response genes known to be downstream of NF{kappa}B tended to have higher serum antibody responses one month after vaccination. A supervised analysis of the same data resulted in a single classifier, also enriched for NF{kappa}B regulated genes, that predicted the antibody response across most of the vaccines. Projection into single-cell RNA-sequencing data suggested that this pre-vaccination state was attributable to the signature of activation of non-classical monocytes and myeloid dendritic cells. Transcriptional signatures of acute responses to bacterial and not viral infections were enriched in the high pro-inflammatory pre-vaccination state and also included NF{kappa}B regulated genes. The pro-inflammatory pre-vaccination state was highly reminiscent of the innate activation state triggered by TLR ligands or adjuvants. These results demonstrate that wide variations in the transcriptional state of the immune system in humans can be a key determinant of responsiveness to vaccination. They also define a transcriptional signature NF{kappa}B activation at baseline, that is associated with a greater magnitude of antibody response to multiple vaccines, and suggest that modulation of the innate immune system by next-generation adjuvants targeting NF{kappa}B before vaccine administration may improve vaccine responsiveness.
]]></description>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Tomalin, L. E.</dc:creator>
<dc:creator>Mule, M. P.</dc:creator>
<dc:creator>Chawla, D. G.</dc:creator>
<dc:creator>Gerritsen, B.</dc:creator>
<dc:creator>Rychkov, D.</dc:creator>
<dc:creator>Henrich, E.</dc:creator>
<dc:creator>Miller, H. E.</dc:creator>
<dc:creator>Hagan, T.</dc:creator>
<dc:creator>Diray-Arce, J.</dc:creator>
<dc:creator>Dunn, P.</dc:creator>
<dc:creator>The Human Immunology Project Consortium (HIPC),</dc:creator>
<dc:creator>Levy, O.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Sarwal, M.</dc:creator>
<dc:creator>Tsang, J.</dc:creator>
<dc:creator>Suarez-Farinas, M.</dc:creator>
<dc:creator>Pulendran, B.</dc:creator>
<dc:creator>Sekaly, R.-P.</dc:creator>
<dc:creator>Kleinstein, S. H.</dc:creator>
<dc:date>2021-09-26</dc:date>
<dc:identifier>doi:10.1101/2021.09.26.461847</dc:identifier>
<dc:title><![CDATA[An innate immune activation state prior to vaccination predicts responsiveness to multiple vaccines]]></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/10.1101/2021.09.29.462325v1?rss=1">
<title>
<![CDATA[
An international report on bacterial communities in esophageal squamous cell carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.29.462325v1?rss=1</link>
<description><![CDATA[
The incidence of esophageal squamous cell carcinoma (ESCC) is disproportionately high in the eastern corridor of Africa and parts of Asia. Emerging research has identified a potential association between poor oral health and ESCC. One proposed biological pathway linking poor oral health and ESCC involves the alteration of the microbiome. Thus, we performed an integrated analysis of four independent sequencing efforts of ESCC tumors from patients from high- and low-incidence regions of the world. Using whole genome sequencing (WGS) and RNA sequencing (RNAseq) of ESCC tumors and WGS of synchronous collections of saliva specimens from 61 patients in Tanzania, we identified a community of bacteria, including members of the genera Fusobacterium, Selenomonas, Prevotella, Streptococcus, Porphyromonas, Veillonella, and Campylobacter, present at high abundance in ESCC tumors. We then characterized the microbiome of 238 ESCC tumor specimens collected in two additional independent sequencing efforts consisting of patients from other high-ESCC incidence regions (Tanzania, Malawi, Kenya, Iran, China). This analysis revealed a similar tumor enrichment of the ESCC-associated bacterial community in these cancers. Because these genera are traditionally considered members of the oral microbiota, we explored if there is a relationship between the synchronous saliva and tumor microbiomes of ESCC patients in Tanzania. Comparative analyses revealed that paired saliva and tumor microbiomes are significantly similar with a specific enrichment of Fusobacterium and Prevotella in the tumor microbiome. Together, these data indicate that cancer-associated oral bacteria are associated with ESCC tumors at the time of diagnosis and support a model in which oral bacteria are present in high abundance in both saliva and tumors of ESCC patients. Longitudinal studies of the pre-diagnostic oral microbiome are needed to investigate whether these cross-sectional similarities reflect temporal associations.
]]></description>
<dc:creator>Nomburg, J.</dc:creator>
<dc:creator>Bullman, S.</dc:creator>
<dc:creator>Nasrollahzadeh, D.</dc:creator>
<dc:creator>Collisson, E. A.</dc:creator>
<dc:creator>Abedi-Ardekani, B.</dc:creator>
<dc:creator>Akoko, L. O.</dc:creator>
<dc:creator>Atkins, J. R.</dc:creator>
<dc:creator>Buckle, G. C.</dc:creator>
<dc:creator>Gopal, S.</dc:creator>
<dc:creator>Hu, N.</dc:creator>
<dc:creator>Kaimila, B.</dc:creator>
<dc:creator>Khoshnia, M.</dc:creator>
<dc:creator>Malekzadeh, R.</dc:creator>
<dc:creator>Menya, D.</dc:creator>
<dc:creator>Mmbaga, B. T.</dc:creator>
<dc:creator>Moody, S.</dc:creator>
<dc:creator>Mulima, G.</dc:creator>
<dc:creator>Mushi, B. P.</dc:creator>
<dc:creator>Mwaiselage, J.</dc:creator>
<dc:creator>Mwanga, A.</dc:creator>
<dc:creator>Newton, Y.</dc:creator>
<dc:creator>Ng, D. L.</dc:creator>
<dc:creator>Radenbaugh, A.</dc:creator>
<dc:creator>Rwakatema, D. S.</dc:creator>
<dc:creator>Selekwa, M.</dc:creator>
<dc:creator>Schüz, J.</dc:creator>
<dc:creator>Taylor, P. R.</dc:creator>
<dc:creator>Vaske, C.</dc:creator>
<dc:creator>Goldstein, A.</dc:creator>
<dc:creator>Stratton, M. R.</dc:creator>
<dc:creator>McCormack, V.</dc:creator>
<dc:creator>Brennan, P.</dc:creator>
<dc:creator>DeCaprio, J. A.</dc:creator>
<dc:creator>Meyerson, M.</dc:creator>
<dc:creator>Mmbaga, E. J.</dc:creator>
<dc:creator>Van Loon, K.</dc:creator>
<dc:date>2021-09-29</dc:date>
<dc:identifier>doi:10.1101/2021.09.29.462325</dc:identifier>
<dc:title><![CDATA[An international report on bacterial communities in esophageal squamous cell carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.30.462231v1?rss=1">
<title>
<![CDATA[
Structures of core eukaryotic protein complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.30.462231v1?rss=1</link>
<description><![CDATA[
Protein-protein interactions play critical roles in biology, but despite decades of effort, the structures of many eukaryotic protein complexes are unknown, and there are likely many interactions that have not yet been identified. Here, we take advantage of recent advances in proteome-wide amino acid coevolution analysis and deep-learning-based structure modeling to systematically identify and build accurate models of core eukaryotic protein complexes, as represented within the Saccharomyces cerevisiae proteome. We use a combination of RoseTTAFold and AlphaFold to screen through paired multiple sequence alignments for 8.3 million pairs of S. cerevisiae proteins and build models for strongly predicted protein assemblies with two to five components. Comparison to existing interaction and structural data suggests that these predictions are likely to be quite accurate. We provide structure models spanning almost all key processes in Eukaryotic cells for 104 protein assemblies which have not been previously identified, and 608 which have not been structurally characterized.

One-sentence summaryWe take advantage of recent advances in proteome-wide amino acid coevolution analysis and deep-learning-based structure modeling to systematically identify and build accurate models of core eukaryotic protein complexes.
]]></description>
<dc:creator>Humphreys, I. R.</dc:creator>
<dc:creator>Pei, J.</dc:creator>
<dc:creator>Baek, M.</dc:creator>
<dc:creator>Krishnakumar, A.</dc:creator>
<dc:creator>Anishchenko, I.</dc:creator>
<dc:creator>Ovchinnikov, S. R.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Ness, T.</dc:creator>
<dc:creator>Banjade, S.</dc:creator>
<dc:creator>Bagde, S. R.</dc:creator>
<dc:creator>Stancheva, V.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Liu, K.</dc:creator>
<dc:creator>Zheng, Z.</dc:creator>
<dc:creator>Barerro, D.</dc:creator>
<dc:creator>Roy, U.</dc:creator>
<dc:creator>Fernandez, I. S.</dc:creator>
<dc:creator>Szakal, B.</dc:creator>
<dc:creator>Branzei, D.</dc:creator>
<dc:creator>Greene, E. C.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:creator>Keeney, S.</dc:creator>
<dc:creator>Miller, E. A.</dc:creator>
<dc:creator>Fromme, J. C.</dc:creator>
<dc:creator>Hendrickson, T.</dc:creator>
<dc:creator>Cong, Q.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2021-09-30</dc:date>
<dc:identifier>doi:10.1101/2021.09.30.462231</dc:identifier>
<dc:title><![CDATA[Structures of core eukaryotic protein complexes]]></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/10.1101/2021.09.30.462547v1?rss=1">
<title>
<![CDATA[
Cx43 carboxyl terminal domain determines AQP4 and Cx30 endfoot organization and blood brain barrier permeability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.30.462547v1?rss=1</link>
<description><![CDATA[
The neurovascular unit (NVU) consists of cells intrinsic to the vessel wall, the endothelial cells and pericytes, and astrocyte endfeet that surround the vessel but are separated from it by basement membrane. Endothelial cells are primarily responsible for creating and maintaining blood-brain-barrier (BBB) tightness, but astrocytes contribute to the barrier through paracrine signaling to the endothelial cells and by forming the glia limitans. Gap junctions (GJs) between astrocyte endfeet are composed of connexin 43 (Cx43) and Cx30, which form plaques between cells. GJ plaques formed of Cx43 do not diffuse laterally in the plasma membrane and thus potentially provide stable organizational features to the endfoot domain, whereas GJ plaques formed of other connexins and of Cx43 lacking a large portion of its cytoplasmic carboxyl terminus are quite mobile. In order to examine the organizational features that immobile GJs impose on the endfoot, we have used super-resolution confocal microscopy to map number and sizes of GJ plaques and aquaporin (AQP)-4 channel clusters in the perivascular endfeet of mice in which astrocyte GJs (Cx30, Cx43) were deleted or the carboxyl terminus of Cx43 was truncated. To determine if blood-brain-barrier integrity was compromised in these transgenic mice, we conducted perfusion studies under elevated hydrostatic pressure using horseradish peroxide as a molecular probe enabling detection of micro-hemorrhages in brain sections. These studies revealed that microhemorrhages were more numerous in mice lacking Cx43 or its carboxyl terminus. In perivascular domains of cerebral vessels, we found that density of Cx43 GJs was higher in the truncation mutant, while GJ size was smaller. Density of perivascular particles formed by AQP4 and its extended isoform AQP4ex was inversely related to the presence of full length Cx43, whereas the ratio of sizes of the particles of the AQP4ex isoform to total AQP4 was directly related to the presence of full length Cx43. Confocal analysis showed that Cx43 and Cx30 were substantially colocalized in astrocyte domains near vasculature of truncation mutant mice. These results showing altered distribution of some astrocyte nexus components (AQP4 and Cx30) in Cx43 null mice and in a truncation mutant, together with leakier cerebral vasculature, support the hypothesis that localization and mobility of gap junction proteins and their binding partners influences organization of astrocyte endfeet which in turn impacts BBB integrity of the NVU.
]]></description>
<dc:creator>Cibelli, A.</dc:creator>
<dc:creator>Stout, R.</dc:creator>
<dc:creator>Timmermann, A.</dc:creator>
<dc:creator>de Menezes, L.</dc:creator>
<dc:creator>Guo, P.</dc:creator>
<dc:creator>Maass, K.</dc:creator>
<dc:creator>Seifert, G.</dc:creator>
<dc:creator>Steinhaeuser, C.</dc:creator>
<dc:creator>Spray, D. C.</dc:creator>
<dc:creator>Scemes, E.</dc:creator>
<dc:date>2021-10-01</dc:date>
<dc:identifier>doi:10.1101/2021.09.30.462547</dc:identifier>
<dc:title><![CDATA[Cx43 carboxyl terminal domain determines AQP4 and Cx30 endfoot organization and blood brain barrier permeability]]></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/10.1101/2021.10.05.463210v1?rss=1">
<title>
<![CDATA[
Comprehensive characterization of the antibody responses to SARS-CoV-2 Spike protein after infection and/or vaccination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.05.463210v1?rss=1</link>
<description><![CDATA[
BackgroundControl of the COVID-19 pandemic will rely on SARS-CoV-2 vaccine-elicited antibodies to protect against emerging and future variants; an understanding of the unique features of the humoral responses to infection and vaccination, including different vaccine platforms, is needed to achieve this goal.

MethodsThe epitopes and pathways of escape for Spike-specific antibodies in individuals with diverse infection and vaccination history were profiled using Phage-DMS. Principal component analysis was performed to identify regions of antibody binding along the Spike protein that differentiate the samples from one another. Within these epitope regions we determined potential escape mutations by comparing antibody binding of peptides containing wildtype residues versus peptides containing a mutant residue.

ResultsIndividuals with mild infection had antibodies that bound to epitopes in the S2 subunit within the fusion peptide and heptad-repeat regions, whereas vaccinated individuals had antibodies that additionally bound to epitopes in the N- and C-terminal domains of the S1 subunit, a pattern that was also observed in individuals with severe disease due to infection. Epitope binding appeared to change over time after vaccination, but other covariates such as mRNA vaccine dose, mRNA vaccine type, and age did not affect antibody binding to these epitopes. Vaccination induced a relatively uniform escape profile across individuals for some epitopes, whereas there was much more variation in escape pathways in in mildly infected individuals. In the case of antibodies targeting the fusion peptide region, which was a common response to both infection and vaccination, the escape profile after infection was not altered by subsequent vaccination.

ConclusionsThe finding that SARS-CoV-2 mRNA vaccination resulted in binding to additional epitopes beyond what was seen after infection suggests protection could vary depending on the route of exposure to Spike antigen. The relatively conserved escape pathways to vaccine-induced antibodies relative to infection-induced antibodies suggests that if escape variants emerge, they may be readily selected for across vaccinated individuals. Given that the majority of people will be first exposed to Spike via vaccination and not infection, this work has implications for predicting the selection of immune escape variants at a population level.

FundingThis work was supported by NIH grants AI138709 (PI Overbaugh) and AI146028 (PI Matsen). Julie Overbaugh received support as the Endowed Chair for Graduate Education (FHCRC). The research of Frederick Matsen was supported in part by a Faculty Scholar grant from the Howard Hughes Medical Institute and the Simons Foundation. Scientific Computing Infrastructure at Fred Hutch was funded by ORIP grant S10OD028685.
]]></description>
<dc:creator>Garrett, M. E.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>Wolf, C.</dc:creator>
<dc:creator>Logue, J. K.</dc:creator>
<dc:creator>Franko, N.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Matsen, F.</dc:creator>
<dc:creator>Overbaugh, J. M.</dc:creator>
<dc:date>2021-10-07</dc:date>
<dc:identifier>doi:10.1101/2021.10.05.463210</dc:identifier>
<dc:title><![CDATA[Comprehensive characterization of the antibody responses to SARS-CoV-2 Spike protein after infection and/or vaccination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.11.463514v1?rss=1">
<title>
<![CDATA[
Whole genome sequence analysis of blood lipid levels in >66,000 individuals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.11.463514v1?rss=1</link>
<description><![CDATA[
Plasma lipids are heritable modifiable causal factors for coronary artery disease, the leading cause of death globally. Despite the well-described monogenic and polygenic bases of dyslipidemia, limitations remain in discovery of lipid-associated alleles using whole genome sequencing, partly due to limited sample sizes, ancestral diversity, and interpretation of potential clinical significance. Increasingly larger whole genome sequence datasets with plasma lipids coupled with methodologic advances enable us to more fully catalog the allelic spectrum for lipids. Here, among 66,329 ancestrally diverse (56% non-European ancestry) participants, we associate 428M variants from deep-coverage whole genome sequences with plasma lipids. Approximately 400M of these variants were not studied in prior lipids genetic analyses. We find multiple lipid-related genes strongly associated with plasma lipids through analysis of common and rare coding variants. We additionally discover several significantly associated rare non-coding variants largely at Mendelian lipid genes. Notably, we detect rare LDLR intronic variants associated with markedly increased LDL-C, similar to rare LDLR exonic variants. In conclusion, we conducted a systematic whole genome scan for plasma lipids expanding the alleles linked to lipids for multiple ancestries and characterize a clinically-relevant rare non-coding variant model for lipids.
]]></description>
<dc:creator>Selvaraj, M. S.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Pampana, A.</dc:creator>
<dc:creator>Zhang, D. Y.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Aslibekyan, S.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Brody, J. A.</dc:creator>
<dc:creator>Cade, B. E.</dc:creator>
<dc:creator>Chuang, L.-M.</dc:creator>
<dc:creator>Chung, R.-H.</dc:creator>
<dc:creator>Curran, J. E.</dc:creator>
<dc:creator>de las Fuentes, L.</dc:creator>
<dc:creator>de Vries, P. S.</dc:creator>
<dc:creator>Duggirala, R.</dc:creator>
<dc:creator>Freedman, B. I.</dc:creator>
<dc:creator>Graff, M.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Heard-Costa, N.</dc:creator>
<dc:creator>Hidalgo, B.</dc:creator>
<dc:creator>Hwu, C.-M.</dc:creator>
<dc:creator>Irvin, M. R.</dc:creator>
<dc:creator>Kelly, T. N.</dc:creator>
<dc:creator>Kral, B. G.</dc:creator>
<dc:creator>Lange, L.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Lisa, M.</dc:creator>
<dc:creator>Lubitz, S. A.</dc:creator>
<dc:creator>Manichaikul, A. W.</dc:creator>
<dc:creator>Michael, P.</dc:creator>
<dc:creator>Montasser, M. E.</dc:creator>
<dc:creator>Morrison, A. C.</dc:creator>
<dc:creator>Naseri, T.</dc:creator>
<dc:creator>O'Connell, J. R.</dc:creator>
<dc:creator>Palmer, N. D.</dc:creator>
<dc:creator>Peyser, P. A.</dc:creator>
<dc:creator>Reupena, M. S.</dc:creator>
<dc:creator>Smith, J. A.</dc:creator>
<dc:creator>Sun, X.</dc:creator>
<dc:creator>Taylor, K. D.</dc:creator>
<dc:creator>Tracy, R. P.</dc:creator>
<dc:creator>Tsai, M. Y.</dc:creator>
<dc:creator>Wang,</dc:creator>
<dc:date>2021-10-12</dc:date>
<dc:identifier>doi:10.1101/2021.10.11.463514</dc:identifier>
<dc:title><![CDATA[Whole genome sequence analysis of blood lipid levels in >66,000 individuals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.12.464112v1?rss=1">
<title>
<![CDATA[
In vivo versus in silico assessment of potentially pathogenic missense variants in human reproductive genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.12.464112v1?rss=1</link>
<description><![CDATA[
Infertility is a heterogeneous condition, with genetic causes estimated to be involved in approximately half of the cases. High-throughput sequencing (HTS) is becoming an increasingly important tool for genetic diagnosis of diseases including idiopathic infertility, however, most rare or minor alleles revealed by HTS are variants of uncertain significance (VUS). Interpreting the functional impacts of VUS is challenging but profoundly important for clinical management and genetic counseling. To determine the consequences of population polymorphisms in key fertility genes, we functionally evaluated 11 missense variants in the genes ANKRD31, BRDT, DMC1, EXOI, FKBP6, MCM9, M1AP, MEI1, MSH4 and SEPT12 by generating genome-edited mouse models. Nine variants were classified as deleterious by most functional prediction algorithms, and two disrupted a protein-protein interaction in the yeast 2 hybrid assay. Even though these genes are known to be essential for normal meiosis or spermiogenesis in mice, only one of the tested human variants (rs1460351219, encoding p.R581H in MCM9), which was observed in a male infertility patient, compromised fertility or gametogenesis in the mouse models. To explore the disconnect between predictions and outcomes, we compared pathogenicity calls of missense variants made by ten widely-used algorithms to: 1) those present in ClinVar, and 2) those which have been evaluated in mice. We found that all the algorithms performed poorly in terms of predicting the effects of human missense variants that have been modeled in mice. These studies emphasize caution in the genetic diagnoses of infertile patients based primarily on pathogenicity prediction algorithms, and emphasize the need for alternative and efficient in vitro or vivo functional validation models for more effective and accurate VUS delineation to either pathogenic or benign categories.

SignificanceAlthough infertility is a substantial medical problem that affects up to 15% of couples, the potential genetic causes of idiopathic infertility have been difficult to decipher. This problem is complicated by the large number of genes that can cause infertility when perturbed, coupled with the large number of VUS that are present in the genomes of affected patients. Here, we present and analyze mouse modeling data of missense variants that are classified as deleterious by commonly-used pathogenicity prediction algorithms but which caused no detectible phenotype when introduced into mice by genome editing. We find that augmenting pathogenicity predictions with preliminary screens for biochemical defects substantially enhanced the proportion of prioritized variants that caused phenotypes in mice. The results emphasize that, in the absence of substantial improvements of in silico prediction tools or other compelling pre-existing evidence, in vivo analysis is crucial for confident attribution of infertility alleles.
]]></description>
<dc:creator>Ding, X.</dc:creator>
<dc:creator>Singh, P.</dc:creator>
<dc:creator>Tran, T. N.</dc:creator>
<dc:creator>Fragoza, R.</dc:creator>
<dc:creator>Yu, H.</dc:creator>
<dc:creator>Schimenti, J. C.</dc:creator>
<dc:date>2021-10-12</dc:date>
<dc:identifier>doi:10.1101/2021.10.12.464112</dc:identifier>
<dc:title><![CDATA[In vivo versus in silico assessment of potentially pathogenic missense variants in human reproductive genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.12.464114v1?rss=1">
<title>
<![CDATA[
A SARS-CoV-2 variant elicits an antibody response with a shifted immunodominance hierarchy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.12.464114v1?rss=1</link>
<description><![CDATA[
Many SARS-CoV-2 variants have mutations at key sites targeted by antibodies. However, it is unknown if antibodies elicited by infection with these variants target the same or different regions of the viral spike as antibodies elicited by earlier viral isolates. Here we compare the specificities of polyclonal antibodies produced by humans infected with early 2020 isolates versus the B.1.351 variant of concern (also known as Beta or 20H/501Y.V2), which contains mutations in multiple key spike epitopes. The serum neutralizing activity of antibodies elicited by infection with both early 2020 viruses and B.1.351 is heavily focused on the spike receptor-binding domain (RBD). However, within the RBD, B.1.351-elicited antibodies are more focused on the "class 3" epitope spanning sites 443 to 452, and neutralization by these antibodies is notably less affected by mutations at residue 484. Our results show that SARS-CoV-2 variants can elicit polyclonal antibodies with different immunodominance hierarchies.
]]></description>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Eguia, R. T.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Khan, K.</dc:creator>
<dc:creator>Karim, F.</dc:creator>
<dc:creator>Cele, S.</dc:creator>
<dc:creator>Bowen, J. E.</dc:creator>
<dc:creator>Logue, J. K.</dc:creator>
<dc:creator>Corti, D.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Sigal, A.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-10-13</dc:date>
<dc:identifier>doi:10.1101/2021.10.12.464114</dc:identifier>
<dc:title><![CDATA[A SARS-CoV-2 variant elicits an antibody response with a shifted immunodominance hierarchy]]></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/10.1101/2021.10.15.464608v1?rss=1">
<title>
<![CDATA[
Kernel-based genetic association analysis for microbiome phenotypes identifies host genetic drivers of beta-diversity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.15.464608v1?rss=1</link>
<description><![CDATA[
Understanding human genetic influences on the gut microbiota helps elucidate the mechanisms by which genetics affects health outcomes. We propose a novel approach, the covariate-adjusted kernel RV (KRV) framework, to map genetic variants associated with microbiome beta-diversity, which focuses on overall shifts in the microbiota. The proposed KRV framework improves statistical power by capturing intrinsic structure within the genetic and microbiome data while reducing the multiple-testing burden. We apply the covariate-adjusted KRV test to the Hispanic Community Health Study/Study of Latinos in a genome-wide association analysis (first gene-level, then variant-level) for microbiome beta-diversity. We have identified an immunity-related gene, IL23R, reported in previous association studies and discovered 3 other novel genes, 2 of which are involved in immune functions or autoimmune disorders. Our findings highlight the value of the KRV as a powerful microbiome GWAS approach and support an important role of immunity-related genes in shaping the gut microbiome composition.
]]></description>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Ling, W.</dc:creator>
<dc:creator>Hua, X.</dc:creator>
<dc:creator>Moon, J.-Y.</dc:creator>
<dc:creator>Williams-Nguyen, J. S.</dc:creator>
<dc:creator>Zhan, X.</dc:creator>
<dc:creator>Plantinga, A. M.</dc:creator>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Zhang, A.</dc:creator>
<dc:creator>Knight, R.</dc:creator>
<dc:creator>Qi, Q.</dc:creator>
<dc:creator>Burk, R. D.</dc:creator>
<dc:creator>Kaplan, R. C.</dc:creator>
<dc:creator>Wu, M. C.</dc:creator>
<dc:date>2021-10-16</dc:date>
<dc:identifier>doi:10.1101/2021.10.15.464608</dc:identifier>
<dc:title><![CDATA[Kernel-based genetic association analysis for microbiome phenotypes identifies host genetic drivers of beta-diversity]]></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/10.1101/2021.10.19.464955v1?rss=1">
<title>
<![CDATA[
Global and context-specific transcriptional consequences of oncogenic Fbw7 mutations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.19.464955v1?rss=1</link>
<description><![CDATA[
Fbw7 is a ubiquitin ligase substrate receptor that targets proteins for proteasomal degradation. Most known Fbw7 substrates are transcription factors (TFs) and many are also oncoproteins (e.g., c-Myc, c-Jun, Notch). Fbw7 is an important tumor suppressor and FBXW7 mutations drive tumorigenesis through activation of oncogenic Fbw7 substrates. Defining the mechanisms of Fbw7-associated tumorigenesis is critical for developing targeted therapies. We thus determined the transcriptional consequences of oncogenic Fbw7 mutations by studying isogenic colorectal cancer cell lines with engineered FBXW7 null and heterozygous missense mutations. We used an integrated approach employing RNA-Seq and high-resolution mapping (CUT&RUN) of histone modifications and TF occupancy (c-Jun and c-Myc) to examine the combinatorial effects of mis-regulated Fbw7 substrates. Fbw7 mutations caused widespread transcriptional changes associated with active chromatin and altered TF occupancy at distal regulatory regions. Some regulatory changes were common to both FBXW7-mutant cell lines whereas others were FBXW7 mutation-specific. By comparing c-Jun and c-Myc binding sites, we also identified co-regulated elements, suggesting that Fbw7 substrates may have synergistic effects. One co-regulated gene was CIITA, a master regulator of MHC Class II gene expression, and Fbw7 loss increased CIITA and MHC Class II gene expression in colorectal cancer cells. Fbw7 mutations were also correlated with increased CIITA expression in TCGA colorectal tumors and cell lines, which may have immunologic implications for progression and treatment of Fbw7-associated cancers. This integrative analysis provides a framework for understanding normal and neoplastic context-specific Fbw7 functions.
]]></description>
<dc:creator>Thirmanne, H. N.</dc:creator>
<dc:creator>Wu, F.</dc:creator>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Swanger, J.</dc:creator>
<dc:creator>Feldman, H.</dc:creator>
<dc:creator>Amezquita, R.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Paddison, P. J.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Clurman, B. E.</dc:creator>
<dc:date>2021-10-21</dc:date>
<dc:identifier>doi:10.1101/2021.10.19.464955</dc:identifier>
<dc:title><![CDATA[Global and context-specific transcriptional consequences of oncogenic Fbw7 mutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.20.465157v1?rss=1">
<title>
<![CDATA[
The endophilin curvature-sensitive motif requires electrostatic guidance to recycle synaptic vesicles in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.20.465157v1?rss=1</link>
<description><![CDATA[
Curvature-sensing mechanisms assist proteins in executing particular actions on various membrane organelles. Here, we investigated the functional specificity of curvature-sensing amphipathic motifs through the study of endophilin, an endocytic protein for synaptic vesicle recycling. We generated chimeric endophilin proteins by replacing the endophilin amphipathic motif H0 with other curvature-sensing amphipathic motifs. We found that the role of amphipathic motifs cannot simply be extrapolated from the identity of their parental proteins. For example, the amphipathic motif of the nuclear pore complex protein NUP133 functionally replaced the synaptic role of endophilin H0. Interestingly, non-functional endophilin chimeras had similar defects - producing fewer synaptic vesicles but more endosomes - indicating that the curvature-sensing motifs in these chimeras have a common deficiency at reforming synaptic vesicles. Finally, we converted non-functional endophilin chimeras into functional proteins by changing the cationic property of amphipathic motifs, setting a precedent for reprogramming the functional specificity of curvature-sensing motifs in vivo.
]]></description>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Dong, Y.</dc:creator>
<dc:creator>Pant, A.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Masserman, L.</dc:creator>
<dc:creator>McLaughlin, R. N.</dc:creator>
<dc:creator>Bai, J.</dc:creator>
<dc:date>2021-10-21</dc:date>
<dc:identifier>doi:10.1101/2021.10.20.465157</dc:identifier>
<dc:title><![CDATA[The endophilin curvature-sensitive motif requires electrostatic guidance to recycle synaptic vesicles in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.22.465520v1?rss=1">
<title>
<![CDATA[
scGAD: single-cell gene associating domain scores for exploratory analysis of scHi-C data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.22.465520v1?rss=1</link>
<description><![CDATA[
SummaryQuantitative tools are needed to leverage the unprecedented resolution of single-cell high-throughput chromatin conformation (scHi-C) data and to integrate it with other single-cell data modalities. We present single-cell gene associating domain (scGAD) scores as a dimension reduction and exploratory analysis tool for scHi-C data. scGAD enables summarization at the gene level while accounting for inherent gene-level genomic biases. Low-dimensional projections with scGAD capture clustering of cells based on their 3D structures. scGAD enables identifying genes with significant chromatin interactions within and between cell types. We further show that scGAD facilitates the integration of scHi-C data with other single-cell data modalities by enabling its projection onto reference low-dimensional embeddings.

AvailabilityscGAD is part of the BandNorm R package at https://sshen82.github.io/BandNorm/articles/scGAD-tutorial.html.

Contactkeles@stat.wisc.edu, yzheng23@fredhutch.org

Supplementary informationSupplementary data are available at Bioinformatics online.
]]></description>
<dc:creator>Shen, S.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Keles, S.</dc:creator>
<dc:date>2021-10-24</dc:date>
<dc:identifier>doi:10.1101/2021.10.22.465520</dc:identifier>
<dc:title><![CDATA[scGAD: single-cell gene associating domain scores for exploratory analysis of scHi-C data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.25.465802v1?rss=1">
<title>
<![CDATA[
Subtype transcriptomic profiling of myeloid cells in Alzheimer Disease brain illustrates the diversity in active microglia phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.25.465802v1?rss=1</link>
<description><![CDATA[
Microglia contribute to Alzheimers Disease (AD) progression and are candidate therapeutic targets. Human microglia exhibit an array of transcriptional phenotypes implying that accurate manipulation of microglial function will require clarity of their molecular states and context dependent regulation. To increase the number of microglia analyzed per subject we employed fluorescence activated nuclei sorting prior to single-nucleus RNA-seq on human prefrontal cortices. We observed microglia phenotypes previously unrecognized in human brain gene expression studies and mapped their transcriptomic relationships by trajectory inference. Three clusters were enriched for endolysosomal pathways, one of which showed differential expression of AD GWAS genes in addition to genes implicated in nucleic acid detection and interferon signaling. Analysis of the "homeostatic" microglia cluster revealed a uniquely AD subcluster. Our study demonstrates the value of deeply profiling microglia to explore the biological implications of microglia transcriptomic diversity.
]]></description>
<dc:creator>Prater, K. E.</dc:creator>
<dc:creator>Green, K. J.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:creator>Chiou, K. L.</dc:creator>
<dc:creator>Heath, L. M.</dc:creator>
<dc:creator>Rose, S.</dc:creator>
<dc:creator>Shojaie, A.</dc:creator>
<dc:creator>Snyder-Mackler, N.</dc:creator>
<dc:creator>Keene, C. D.</dc:creator>
<dc:creator>Blue, E.</dc:creator>
<dc:creator>Young, J. E.</dc:creator>
<dc:creator>Logsdon, B.</dc:creator>
<dc:creator>Garden, G. A.</dc:creator>
<dc:creator>Jayadev, S.</dc:creator>
<dc:date>2021-10-26</dc:date>
<dc:identifier>doi:10.1101/2021.10.25.465802</dc:identifier>
<dc:title><![CDATA[Subtype transcriptomic profiling of myeloid cells in Alzheimer Disease brain illustrates the diversity in active microglia phenotypes]]></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/10.1101/2021.10.26.465098v1?rss=1">
<title>
<![CDATA[
Adenovirus-α-defensin complexes induce NLRP3-associated maturation of human phagocytes via TLR4 engagement 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.26.465098v1?rss=1</link>
<description><![CDATA[
Intramuscular delivery of human adenovirus (HAdV)-based vaccines leads to rapid recruitment of neutrophils, which then release antimicrobial peptides/proteins (AMPs). How these AMPs influence vaccine efficacy over the subsequent 24 hr is poorly understood. In this study, we asked if human neutrophil protein 1 (HNP-1), an -defensin that influences the direct and indirect innate immune responses to a range of pathogens, impacts the innate response of human phagocytes to three HAdV species/types (HAdV-C5, -D26, -B35). We show that HNP-1 binds to the capsids, redirects HAdV-C5, -D26, -B35 to Toll-like receptor 4 (TLR4), which leads to internalization, an NLRP3-mediated inflammasome response, and IL-1{beta} release. Surprisingly, IL-1{beta} release was not associated with notable disruption of plasma membrane integrity. These data further our understanding of HAdV innate immunogenicity and may provide pathways to extend HAdV-based vaccines efficacy.
]]></description>
<dc:creator>Eichholz, K.</dc:creator>
<dc:creator>Tran, H. T.</dc:creator>
<dc:creator>Cheneau, C.</dc:creator>
<dc:creator>Tran, T. T. P.</dc:creator>
<dc:creator>Paris, O.</dc:creator>
<dc:creator>Pugniere, M.</dc:creator>
<dc:creator>Kremer, E. J.</dc:creator>
<dc:date>2021-10-26</dc:date>
<dc:identifier>doi:10.1101/2021.10.26.465098</dc:identifier>
<dc:title><![CDATA[Adenovirus-α-defensin complexes induce NLRP3-associated maturation of human phagocytes via TLR4 engagement]]></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/10.1101/2021.11.01.466599v1?rss=1">
<title>
<![CDATA[
Multilineage plasticity in prostate cancer through expansion of stem-like luminal epithelial cells with elevated inflammatory signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.01.466599v1?rss=1</link>
<description><![CDATA[
Lineage plasticity is a well-established mechanism of resistance to targeted therapies in lung and prostate cancer, where tumors transition from adenocarcinoma to small-cell or neuroendocrine carcinoma. Through single-cell analysis of a cohort of heavily-treated castration-resistant human prostate cancers (CRPC), we report a greater degree of plasticity than previously appreciated, with multiple distinct neuroendocrine (NEPC), mesenchymal (EMT-like), and other subpopulations detected within single biopsies. To explore the steps leading to this plasticity, we turned to two genetically engineered mouse models of prostate cancer that recapitulate progression from adenocarcinoma to neuroendocrine disease. Time course studies reveal expansion of stem-like luminal epithelial cells (Sca1+, Psca+, called L2) that, based on trajectories, gave rise to at least 4 distinct subpopulations, NEPC (Ascl1+), POU2F3 (Pou2f3+), TFF3 (Tff3+) and EMT-like (Vim+, Ncam1+)--these populations are also seen in human prostate and small cell lung cancers. Transformed L2-like cells express stem-like and gastrointestinal endoderm-like transcriptional programs, indicative of reemerging developmental plasticity programs, as well as elevated Jak/Stat and interferon pathway signaling. In sum, while the magnitude of multilineage heterogeneity, both within and across patients, raises considerable treatment challenges, the identification of highly plastic luminal cells as the likely source of this heterogeneity provides a target for more focused therapeutic intervention.

One Sentence SummaryMultilineage plasticity results from expansion of stem-like luminal cells with JAK/STAT activation, serving as a therapeutic target.
]]></description>
<dc:creator>Zaidi, S.</dc:creator>
<dc:creator>Zhao, J. L.</dc:creator>
<dc:creator>Chan, J. M.</dc:creator>
<dc:creator>Roudier, M. P.</dc:creator>
<dc:creator>Wadosky, K. M.</dc:creator>
<dc:creator>Gopalan, A.</dc:creator>
<dc:creator>Karthaus, W. R.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Lawrence, K.</dc:creator>
<dc:creator>Chaudhary, O.</dc:creator>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Masilionis, I.</dc:creator>
<dc:creator>Mazutis, L.</dc:creator>
<dc:creator>Chaligne, R.</dc:creator>
<dc:creator>Linkov, I.</dc:creator>
<dc:creator>Barlas, A.</dc:creator>
<dc:creator>Jungbluth, A.</dc:creator>
<dc:creator>Rekhtman, N.</dc:creator>
<dc:creator>Silber, J.</dc:creator>
<dc:creator>Manova Todorova, K.</dc:creator>
<dc:creator>Watson, P. A.</dc:creator>
<dc:creator>True, L. D.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Scher, H. I.</dc:creator>
<dc:creator>Rathkopf, D. E.</dc:creator>
<dc:creator>Morris, M. J.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Goodrich, D. W.</dc:creator>
<dc:creator>Pe'er, D.</dc:creator>
<dc:creator>Sawyers, C. L.</dc:creator>
<dc:date>2021-11-03</dc:date>
<dc:identifier>doi:10.1101/2021.11.01.466599</dc:identifier>
<dc:title><![CDATA[Multilineage plasticity in prostate cancer through expansion of stem-like luminal epithelial cells with elevated inflammatory signaling]]></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/10.1101/2021.11.01.466615v1?rss=1">
<title>
<![CDATA[
Reversal of lineage plasticity in RB1/TP53-deleted prostate cancer through FGFR and Janus kinase inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.01.466615v1?rss=1</link>
<description><![CDATA[
The inherent plasticity of tumor cells provides a mechanism of resistance to many molecularly targeted therapies, exemplified by adeno-to-neuroendocrine lineage transitions seen in prostate and lung cancer. Here we investigate the root cause of this lineage plasticity in a primary murine prostate organoid model that mirrors the lineage transition seen in patients. These cells lose luminal identity within weeks following deletion of Trp53 and Rb1, ultimately acquiring an Ar-negative, Syp+ phenotype after orthotopic in vivo transplantation. Single-cell transcriptomic analysis revealed progressive mixing of luminal-basal lineage features after tumor suppressor gene deletion, accompanied by activation of Jak/Stat and Fgfr pathway signaling and interferon-a and -g gene expression programs prior to any morphologic changes. Genetic or pharmacologic inhibition of Jak1/2 in combination with FGFR blockade restored luminal differentiation and sensitivity to antiandrogen therapy in models with residual AR expression. Collectively, we show lineage plasticity initiates quickly as a largely cell-autonomous process and, through newly developed computational approaches, identify a pharmacological strategy that restores lineage identity using clinical grade inhibitors.
]]></description>
<dc:creator>Karthaus, W.</dc:creator>
<dc:creator>Chan, J.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:creator>Love, J.</dc:creator>
<dc:creator>Zaidi, S.</dc:creator>
<dc:creator>Choo, Z.-N.</dc:creator>
<dc:creator>Persad, S.</dc:creator>
<dc:creator>LaClair, J.</dc:creator>
<dc:creator>Lawrence, K.</dc:creator>
<dc:creator>Chaudhary, O.</dc:creator>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Masilionis, I.</dc:creator>
<dc:creator>Mazutis, L.</dc:creator>
<dc:creator>Chaligne, R.</dc:creator>
<dc:creator>Pe'er, D.</dc:creator>
<dc:creator>Sawyers, C.</dc:creator>
<dc:date>2021-11-01</dc:date>
<dc:identifier>doi:10.1101/2021.11.01.466615</dc:identifier>
<dc:title><![CDATA[Reversal of lineage plasticity in RB1/TP53-deleted prostate cancer through FGFR and Janus kinase inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.05.465336v1?rss=1">
<title>
<![CDATA[
The Immune Signatures Data Resource: A compendium of systems vaccinology datasets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.05.465336v1?rss=1</link>
<description><![CDATA[
Vaccines are among the most cost-effective public health interventions for preventing infection-induced morbidity and mortality, yet much remains to be learned regarding the mechanisms by which vaccines protect. Systems immunology combines traditional immunology with modern  omic profiling techniques and computational modeling to promote rapid and transformative advances in vaccinology and vaccine discovery. The NIH/NIAID Human Immunology Project Consortium (HIPC) has leveraged systems immunology approaches to identify molecular signatures associated with the immunogenicity of many vaccines, including those targeting seasonal influenza, yellow fever, and hepatitis B. These data are made available to the broader scientific community through the ImmuneSpace data portal and analysis engine leveraging the NIH/NIAID ImmPort repository1,2. However, a barrier to progress in this area is that comparative analyses have been limited by the distributed nature of some data, potential batch effects across studies, and the absence of multiple relevant studies from non-HIPC groups in ImmPort. To support comparative analyses across different vaccines, we have created the Immune Signatures Data Resource, a compendium of standardized systems vaccinology datasets. This data resource is available through ImmuneSpace, along with code to reproduce the processing and batch normalization starting from the underlying study data in ImmPort and the Gene Expression Omnibus (GEO). The current release comprises 1405 participants from 53 cohorts profiling the response to 24 different vaccines and includes transcriptional profiles and antibody response measurements. This novel systems vaccinology data release represents a valuable resource for comparative and meta-analyses that will accelerate our understanding of mechanisms underlying vaccine responses.


[Table 1]
]]></description>
<dc:creator>Diray-Arce, J.</dc:creator>
<dc:creator>Miller, H. E. R.</dc:creator>
<dc:creator>Henrich, E.</dc:creator>
<dc:creator>Gerritsen, B.</dc:creator>
<dc:creator>Mule, M. P.</dc:creator>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Gygi, J.</dc:creator>
<dc:creator>Hagan, T.</dc:creator>
<dc:creator>Tomalin, L.</dc:creator>
<dc:creator>Rychov, D.</dc:creator>
<dc:creator>Kazmin, D.</dc:creator>
<dc:creator>Chawla, D. G.</dc:creator>
<dc:creator>Meng, H.</dc:creator>
<dc:creator>Dunn, P.</dc:creator>
<dc:creator>Campbell, J.</dc:creator>
<dc:creator>The Human Immunology Project Consortium (HIPC),</dc:creator>
<dc:creator>Sarwal, M.</dc:creator>
<dc:creator>Tsang, J.</dc:creator>
<dc:creator>Levy, O.</dc:creator>
<dc:creator>Pulendran, B.</dc:creator>
<dc:creator>Sekaly, R.-P.</dc:creator>
<dc:creator>Floratos, A.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Kleinstein, S.</dc:creator>
<dc:creator>Suarez-Farinas, M.</dc:creator>
<dc:date>2021-11-08</dc:date>
<dc:identifier>doi:10.1101/2021.11.05.465336</dc:identifier>
<dc:title><![CDATA[The Immune Signatures Data Resource: A compendium of systems vaccinology datasets]]></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/10.1101/2021.11.05.467531v1?rss=1">
<title>
<![CDATA[
A framework for detecting noncoding rare variant associations of large-scale whole-genome sequencing studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.05.467531v1?rss=1</link>
<description><![CDATA[
Large-scale whole-genome sequencing studies have enabled analysis of noncoding rare variants (RVs) associations with complex human traits. Variant set analysis is a powerful approach to study RV association, and a key component of it is constructing RV sets for analysis. However, existing methods have limited ability to define analysis units in the noncoding genome. Furthermore, there is a lack of robust pipelines for comprehensive and scalable noncoding RV association analysis. Here we propose a computationally-efficient noncoding RV association-detection framework that uses STAAR (variant-set test for association using annotation information) to group noncoding variants in gene-centric analysis based on functional categories. We also propose SCANG (scan the genome)-STAAR, which uses dynamic window sizes and incorporates multiple functional annotations, in a non-gene-centric analysis. We furthermore develop STAARpipeline to perform flexible noncoding RV association analysis, including gene-centric analysis as well as fixed-window-based and dynamic-window-based non-gene-centric analysis. We apply STAARpipeline to identify noncoding RV sets associated with four quantitative lipid traits in 21,015 discovery samples from the Trans-Omics for Precision Medicine (TOPMed) program and replicate several noncoding RV associations in an additional 9,123 TOPMed samples.
]]></description>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Zhou, H.</dc:creator>
<dc:creator>Gaynor, S. M.</dc:creator>
<dc:creator>Selvaraj, M. S.</dc:creator>
<dc:creator>Arapoglou, T.</dc:creator>
<dc:creator>Quick, C.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Sun, R.</dc:creator>
<dc:creator>Dey, R.</dc:creator>
<dc:creator>Arnett, D. K.</dc:creator>
<dc:creator>Bielak, L. F.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Blackwell, T.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Boerwinkle, E.</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>Conomos, M.</dc:creator>
<dc:creator>Correa, A.</dc:creator>
<dc:creator>Cupples, L. A.</dc:creator>
<dc:creator>Curran, J. E.</dc:creator>
<dc:creator>de Vries, P. S.</dc:creator>
<dc:creator>Duggirala, R.</dc:creator>
<dc:creator>Freedman, B. I.</dc:creator>
<dc:creator>Goring, H. H. H.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Kalyani, R. R.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Kral, B. G.</dc:creator>
<dc:creator>Lange, L. A.</dc:creator>
<dc:creator>Manichaikul, A. W.</dc:creator>
<dc:creator>Martin, L. W.</dc:creator>
<dc:creator>Mitchell, B. D.</dc:creator>
<dc:creator>Montasser, M.</dc:creator>
<dc:creator>Morrison, A.</dc:creator>
<dc:creator>Naseri, T.</dc:creator>
<dc:creator>O'Connell, J. R.</dc:creator>
<dc:creator>Palmer, N. D.</dc:creator>
<dc:creator>Peyser, P. A.</dc:creator>
<dc:creator>Psaty, B. M.</dc:creator>
<dc:creator>Raff</dc:creator>
<dc:date>2021-11-08</dc:date>
<dc:identifier>doi:10.1101/2021.11.05.467531</dc:identifier>
<dc:title><![CDATA[A framework for detecting noncoding rare variant associations of large-scale whole-genome sequencing studies]]></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/10.1101/2021.11.11.468255v1?rss=1">
<title>
<![CDATA[
Computational design of nanoscale rotational mechanics in de novo protein assemblies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.11.468255v1?rss=1</link>
<description><![CDATA[
Natural nanomachines like the F1/F0-ATPase contain protein components that undergo rotation relative to each other. Designing such mechanically constrained nanoscale protein architectures with internal degrees of freedom is an outstanding challenge for computational protein design. Here we explore the de novo construction of protein rotary machinery from designed axle and ring components. Using cryoelectron microscopy, we find that axle-ring systems assemble as designed and populate diverse rotational states depending on symmetry match or mismatch and the designed interface energy landscape. These mechanical systems with internal rotational degrees of freedom are a step towards the systematic design of genetically encodable nanomachines.

One-Sentence SummaryComputationally designed self-assembling protein rotary machines sample internal degrees of freedom sculpted within the energy landscape.
]]></description>
<dc:creator>Courbet, A.</dc:creator>
<dc:creator>Hansen, J. P.</dc:creator>
<dc:creator>Hsia, Y.</dc:creator>
<dc:creator>Bethel, N.</dc:creator>
<dc:creator>Park, Y.-J.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Moyer, A.</dc:creator>
<dc:creator>Boyken, S. E.</dc:creator>
<dc:creator>Ueda, G.</dc:creator>
<dc:creator>Nattermann, U.</dc:creator>
<dc:creator>Nagarajan, D.</dc:creator>
<dc:creator>Silva, D.</dc:creator>
<dc:creator>Sheffler, W.</dc:creator>
<dc:creator>Quispe, J.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Kollman, J. M.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2021-11-12</dc:date>
<dc:identifier>doi:10.1101/2021.11.11.468255</dc:identifier>
<dc:title><![CDATA[Computational design of nanoscale rotational mechanics in de novo protein assemblies]]></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/10.1101/2021.11.18.469171v1?rss=1">
<title>
<![CDATA[
Cell Painting predicts impact of lung cancer variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.18.469171v1?rss=1</link>
<description><![CDATA[
Most variants in most genes across most organisms have an unknown impact on the function of the corresponding gene. This gap in knowledge is especially acute in cancer, where clinical sequencing of tumors now routinely reveals patient-specific variants whose functional impact on the corresponding gene is unknown, impeding clinical utility. Transcriptional profiling was able to systematically distinguish these variants of unknown significance (VUS) as impactful vs. neutral in an approach called expression-based variant-impact phenotyping (eVIP). We profiled a set of lung adenocarcinoma-associated somatic variants using Cell Painting, a morphological profiling assay that captures features of cells based on microscopy using six stains of cell and organelle components. Using deep-learning-extracted features from each cells image, we found that cell morphological profiling (cmVIP) can predict variants functional impact and, particularly at the single-cell level, reveals biological insights into variants which can be explored in our public online portal. Given its low cost, convenient implementation, and single-cell resolution, cmVIP profiling therefore seems promising as an avenue for using non-gene-specific assays to systematically assess the impact of variants, including disease-associated alleles, on gene function.
]]></description>
<dc:creator>Caicedo, J. C.</dc:creator>
<dc:creator>Arevalo, J.</dc:creator>
<dc:creator>Piccioni, F.</dc:creator>
<dc:creator>Bray, M.-A.</dc:creator>
<dc:creator>Hartland, C. L.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Brooks, A. N.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:creator>Boehm, J. S.</dc:creator>
<dc:creator>Carpenter, A. E.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:date>2021-11-20</dc:date>
<dc:identifier>doi:10.1101/2021.11.18.469171</dc:identifier>
<dc:title><![CDATA[Cell Painting predicts impact of lung cancer variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.22.469479v1?rss=1">
<title>
<![CDATA[
Early age-related atrophy of cutaneous lymph nodes precipitates an early functional decline in skin immunity in mice with aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.22.469479v1?rss=1</link>
<description><![CDATA[
Secondary lymphoid organs (SLO; including the spleen and lymph nodes) are critical both for the maintenance of naive T (TN) lymphocytes and for the initiation and coordination of immune responses. How they age, including the exact timing, extent, physiological relevance, and the nature of age-related changes, remains incompletely understood. We used time-stamping to indelibly mark cohorts of newly generated naive T cells (a.k.a. recent thymic emigrants - RTE) in mice, and followed their presence, phenotype and retention in SLO. We found that SLO involute asynchronously. Skin-draining lymph nodes (LN) atrophied early (6-9 months) in life and deeper tissue-draining LN and the spleen late (18-20 months), as measured by the loss of both TN numbers and the fibroblastic reticular cell (FRC) network. Time-stamped RTE cohorts of all ages entered SLO and successfully completed post-thymic differentiation. However, in older mice, these cells were poorly retained, and those found in SLO exhibited an emigration phenotype (CCR7loS1P1hi). Transfers of adult RTE into recipients of different ages formally demonstrated that the defect segregates with the age of the SLO microenvironment and not with the age of T cells. Finally, upon intradermal immunization, RTE generated in mice as early as 6-7 months of age barely participated in de novo immune responses and failed to produce well-armed effector cells. These results highlight changes in structure and function of superficial secondary lymphoid organs in laboratory mice that are earlier than expected and are consistent with the long-appreciated and pronounced reduction of cutaneous immunity with aging.
]]></description>
<dc:creator>Sonar, S. A.</dc:creator>
<dc:creator>Uhrlaub, J. L.</dc:creator>
<dc:creator>Coplen, C. P.</dc:creator>
<dc:creator>Sempowski, G. D.</dc:creator>
<dc:creator>Dudakov, J. A.</dc:creator>
<dc:creator>van den Brink, M. R. M.</dc:creator>
<dc:creator>LaFleur, B. J.</dc:creator>
<dc:creator>Jergovic, M.</dc:creator>
<dc:creator>Nikolich-Zugich, J.</dc:creator>
<dc:date>2021-11-23</dc:date>
<dc:identifier>doi:10.1101/2021.11.22.469479</dc:identifier>
<dc:title><![CDATA[Early age-related atrophy of cutaneous lymph nodes precipitates an early functional decline in skin immunity in mice with aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.01.470697v1?rss=1">
<title>
<![CDATA[
Macaque-human differences in SARS-CoV-2 Spike antibody response elicited by vaccination or infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.01.470697v1?rss=1</link>
<description><![CDATA[
Macaques are a commonly used model for studying immunity to human viruses, including for studies of SARS-CoV-2 infection and vaccination. However, it is unknown whether macaque antibody responses recapitulate, and thus appropriately model, the response in humans. To answer this question, we employed a phage-based deep mutational scanning approach (Phage- DMS) to compare which linear epitopes are targeted on the SARS-CoV-2 Spike protein in humans and macaques following either vaccination or infection. We also used Phage-DMS to determine antibody escape pathways within each epitope, enabling a granular comparison of antibody binding specificities at the locus level. Overall, we identified some common epitope targets in both macaques and humans, including in the fusion peptide (FP) and stem helix- heptad repeat 2 (SH-H) regions. Differences between groups included a response to epitopes in the N-terminal domain (NTD) and C-terminal domain (CTD) in vaccinated humans but not vaccinated macaques, as well as recognition of a CTD epitope and epitopes flanking the FP in convalescent macaques but not convalescent humans. There was also considerable variability in the escape pathways among individuals within each group. Sera from convalescent macaques showed the least variability in escape overall and converged on a common response with vaccinated humans in the SH-H epitope region, suggesting highly similar antibodies were elicited. Collectively, these findings suggest that the antibody response to SARS-CoV-2 in macaques shares many features with humans, but with substantial differences in the recognition of certain epitopes and considerable individual variability in antibody escape profiles, suggesting a diverse repertoire of antibodies that can respond to major epitopes in both humans and macaques.

Author summaryNon-human primates, including macaques, are considered the best animal model for studying infectious diseases that infect humans. Vaccine candidates for SARS-CoV-2 are first tested in macaques to assess immune responses prior to advancing to human trials, and macaques are also used to model the human immune response to SARS-CoV-2 infection. However, there may be differences in how macaque and human antibodies recognize the SARS-CoV-2 entry protein, Spike. Here we characterized the locations on Spike that are recognized by antibodies from vaccinated or infected macaques and humans. We also made mutations to the viral sequence and assessed how these affected antibody binding, enabling a comparison of antibody binding requirements between macaques and humans at a very precise level. We found that macaques and humans share some responses, but also recognize distinct regions of Spike. We also found that in general, antibodies from different individuals had unique responses to viral mutations, regardless of species. These results will yield a better understanding of how macaque data can be used to inform human immunity to SARS-CoV-2.
]]></description>
<dc:creator>Willcox, A.</dc:creator>
<dc:creator>Sung, K.</dc:creator>
<dc:creator>Garrett, M. E.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>OConnor, M. A.</dc:creator>
<dc:creator>Erasmus, J. H.</dc:creator>
<dc:creator>Logue, J. K.</dc:creator>
<dc:creator>Hawman, D. W.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Hasenkrug, K. J.</dc:creator>
<dc:creator>Fuller, D. H.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2021-12-03</dc:date>
<dc:identifier>doi:10.1101/2021.12.01.470697</dc:identifier>
<dc:title><![CDATA[Macaque-human differences in SARS-CoV-2 Spike antibody response elicited by vaccination or infection]]></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/10.1101/2021.12.01.470820v1?rss=1">
<title>
<![CDATA[
A Co-abundant Gene Analysis for Identification of Gut Microbial Strains Associated with Immune Responses to Oral Cholera Vaccine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.01.470820v1?rss=1</link>
<description><![CDATA[
The degree of protection conferred after receiving an oral cholera vaccine (OCV) varies based on age, prior exposure to Vibrio cholerae, and unknown factors. Recent evidence suggests that the microbiota may mediate some of the unexplained differences in oral vaccine responses. We used metagenomic sequencing of the microbiota at the time of vaccination, and then related microbial features to immune responses after OCV using a reference-independent gene-level analysis. We found that the presence of sphingolipid-producing bacteria was associated with the development of protective immune responses after OCV. We experimentally tested these results by stimulating human macrophages with Bacteroides xylanisolvens metabolites and found that sphingolipid-containing extracts increased innate immune responses to OCV antigens. Our findings demonstrate a new analytic method for translating metagenomic sequencing data into strain-specific results associated with a biological outcome, and in validating this tool, we identified that microbe-derived sphingolipids impact in vitro immune responses to OCV antigens.
]]></description>
<dc:creator>Heller, F. J.</dc:creator>
<dc:creator>Banna, H. A.</dc:creator>
<dc:creator>Kaisar, M. H.</dc:creator>
<dc:creator>Chac, D.</dc:creator>
<dc:creator>Chowdhury, F.</dc:creator>
<dc:creator>Bhuiyan, T. R.</dc:creator>
<dc:creator>Akter, A.</dc:creator>
<dc:creator>Khan, A. I.</dc:creator>
<dc:creator>Karmakar, P. C.</dc:creator>
<dc:creator>Dash, P.</dc:creator>
<dc:creator>LaRocque, R. C.</dc:creator>
<dc:creator>Ryan, E. T.</dc:creator>
<dc:creator>Minot, S. S.</dc:creator>
<dc:creator>Harris, J. B.</dc:creator>
<dc:creator>Qadri, F.</dc:creator>
<dc:creator>Weil, A. A.</dc:creator>
<dc:date>2021-12-01</dc:date>
<dc:identifier>doi:10.1101/2021.12.01.470820</dc:identifier>
<dc:title><![CDATA[A Co-abundant Gene Analysis for Identification of Gut Microbial Strains Associated with Immune Responses to Oral Cholera Vaccine]]></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/10.1101/2021.12.04.471236v1?rss=1">
<title>
<![CDATA[
An antibody-escape calculator for mutations to the SARS-CoV-2 receptor-binding domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.04.471236v1?rss=1</link>
<description><![CDATA[
A key goal of SARS-CoV-2 surveillance is to rapidly identify viral variants with mutations that reduce neutralization by polyclonal antibodies elicited by vaccination or infection. Unfortunately, direct experimental characterization of new viral variants lags their sequence-based identification. Here we help address this challenge by aggregating deep mutational scanning data into an "escape calculator" that estimates the antigenic effects of arbitrary combinations of mutations to the viruss spike receptor-binding domain (RBD). The calculator can be used to intuitively visualize how mutations impact polyclonal antibody recognition, and score the expected antigenic effect of combinations of mutations. These scores correlate with neutralization assays performed on SARS-CoV-2 variants, and emphasize the ominous antigenic properties of the recently described Omicron variant. An interactive version of the calculator is at https://jbloomlab.github.io/SARS2_RBD_Ab_escape_maps/escape-calc/, and we provide a Python module for batch processing.
]]></description>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2021-12-07</dc:date>
<dc:identifier>doi:10.1101/2021.12.04.471236</dc:identifier>
<dc:title><![CDATA[An antibody-escape calculator for mutations to the SARS-CoV-2 receptor-binding domain]]></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/10.1101/2021.12.07.470215v1?rss=1">
<title>
<![CDATA[
A multi-layer functional genomic analysis to understand noncoding genetic variation in lipids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.07.470215v1?rss=1</link>
<description><![CDATA[
A major challenge of genome-wide association studies (GWAS) is to translate phenotypic associations into biological insights. Here, we integrate a large GWAS on blood lipids involving 1.6 million individuals from five ancestries with a wide array of functional genomic datasets to discover regulatory mechanisms underlying lipid associations. We first prioritize lipid-associated genes with expression quantitative trait locus (eQTL) colocalizations, and then add chromatin interaction data to narrow the search for functional genes. Polygenic enrichment analysis across 697 annotations from a host of tissues and cell types confirms the central role of the liver in lipid levels, and highlights the selective enrichment of adipose-specific chromatin marks in high-density lipoprotein cholesterol and triglycerides. Overlapping transcription factor (TF) binding sites with lipid-associated loci identifies TFs relevant in lipid biology. In addition, we present an integrative framework to prioritize causal variants at GWAS loci, producing a comprehensive list of candidate causal genes and variants with multiple layers of functional evidence. Two prioritized genes, CREBRF and RRBP1, show convergent evidence across functional datasets supporting their roles in lipid biology.
]]></description>
<dc:creator>Ramdas, S.</dc:creator>
<dc:creator>Judd, J.</dc:creator>
<dc:creator>Graham, S. E.</dc:creator>
<dc:creator>Kanoni, S.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Surakka, I.</dc:creator>
<dc:creator>Wenz, B.</dc:creator>
<dc:creator>Clarke, S. L.</dc:creator>
<dc:creator>Chesi, A.</dc:creator>
<dc:creator>Wells, A.</dc:creator>
<dc:creator>Bhatti, K. F.</dc:creator>
<dc:creator>Vedantam, S.</dc:creator>
<dc:creator>Winkler, T. W.</dc:creator>
<dc:creator>Locke, A. E.</dc:creator>
<dc:creator>Marouli, E.</dc:creator>
<dc:creator>Zajac, G. J.</dc:creator>
<dc:creator>Wu, K.-H. H.</dc:creator>
<dc:creator>Ntalla, I.</dc:creator>
<dc:creator>Hui, Q.</dc:creator>
<dc:creator>Klarin, D.</dc:creator>
<dc:creator>Hilliard, A. T.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Xue, C.</dc:creator>
<dc:creator>Thorleifsson, G.</dc:creator>
<dc:creator>Helgadottir, A.</dc:creator>
<dc:creator>Gudbjartsson, D. F.</dc:creator>
<dc:creator>Holm, H.</dc:creator>
<dc:creator>Olafsson, I.</dc:creator>
<dc:creator>Hwang, M. Y.</dc:creator>
<dc:creator>Han, S.</dc:creator>
<dc:creator>Akiyama, M.</dc:creator>
<dc:creator>Sakaue, S.</dc:creator>
<dc:creator>Terao, C.</dc:creator>
<dc:creator>Kanai, M.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Brumpton, B. M.</dc:creator>
<dc:creator>Rasheed, H.</dc:creator>
<dc:creator>Havulinna, A. S.</dc:creator>
<dc:creator>Veturi, Y.</dc:creator>
<dc:creator>Pacheco, J. A.</dc:creator>
<dc:creator>Rosenthal, E. A.</dc:creator>
<dc:creator>Lingren, T.</dc:creator>
<dc:creator>Feng, Q.</dc:creator>
<dc:creator>Kullo, I. J.</dc:creator>
<dc:creator>Narita, A.</dc:creator>
<dc:creator>Takayama,</dc:creator>
<dc:date>2021-12-08</dc:date>
<dc:identifier>doi:10.1101/2021.12.07.470215</dc:identifier>
<dc:title><![CDATA[A multi-layer functional genomic analysis to understand noncoding genetic variation in lipids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.08.471817v1?rss=1">
<title>
<![CDATA[
The structural basis of mRNA recognition and binding by eukaryotic pseudouridine synthase PUS1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.08.471817v1?rss=1</link>
<description><![CDATA[
The chemical modification of RNA bases represents a ubiquitous activity that spans all domains of life. Pseudouridylation is the most common RNA modification and is observed within tRNA, rRNA, ncRNA and mRNAs. Pseudouridine synthase or  PUS enzymes include those that rely on guide RNA molecules and others that function as  stand-alone enzymes. Among the latter, several have been shown to modify mRNA transcripts. Although recent studies have defined the structural requirements for RNA to act as a PUS target, the mechanisms by which PUS1 recognizes these target sequences in mRNA are not well understood. Here we describe the crystal structure of yeast PUS1 bound to an RNA target that we identified as being a hot spot for PUS1-interaction within a model mRNA at 2.4 [A] resolution. The enzyme recognizes and binds both strands in a helical base-paired RNA duplex, and thus guides the RNA containing the target uridine to the active site for subsequent modification of the transcript. The study also allows us to show the divergence of related PUS1 enzymes and their corresponding RNA target specificities, and to speculate on the basis by which PUS1 binds and modifies mRNA or tRNA substrates.
]]></description>
<dc:creator>Gruenberg, S.</dc:creator>
<dc:creator>Doyle, L. A.</dc:creator>
<dc:creator>Dai, N.</dc:creator>
<dc:creator>Correa, I. R.</dc:creator>
<dc:creator>Yigit, E.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:date>2021-12-08</dc:date>
<dc:identifier>doi:10.1101/2021.12.08.471817</dc:identifier>
<dc:title><![CDATA[The structural basis of mRNA recognition and binding by eukaryotic pseudouridine synthase PUS1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.10.471810v1?rss=1">
<title>
<![CDATA[
Clonal hematopoiesis is driven by aberrant activation of TCL1A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.10.471810v1?rss=1</link>
<description><![CDATA[
A diverse set of driver genes, such as regulators of DNA methylation, RNA splicing, and chromatin remodeling, have been associated with pre-malignant clonal expansion of hematopoietic stem cells (HSCs). The factors mediating expansion of these mutant clones remain largely unknown, partially due to a paucity of large cohorts with longitudinal blood sampling. To circumvent this limitation, we developed and validated a method to infer clonal expansion rate from single timepoint data called PACER (passenger-approximated clonal expansion rate). Applying PACER to 5,071 persons with clonal hematopoiesis accurately recapitulated the known fitness effects due to different driver mutations. A genome-wide association study of PACER revealed that a common inherited polymorphism in the TCL1A promoter was associated with slower clonal expansion. Those carrying two copies of this protective allele had up to 80% reduced odds of having driver mutations in TET2, ASXL1, SF3B1, SRSF2, and JAK2, but not DNMT3A. TCL1A was not expressed in normal or DNMT3A-mutated HSCs, but the introduction of mutations in TET2 or ASXL1 by CRISPR editing led to aberrant expression of TCL1A and expansion of HSCs in vitro. These effects were abrogated in HSCs from donors carrying the protective TCL1A allele. Our results indicate that the fitness advantage of multiple common driver genes in clonal hematopoiesis is mediated through TCL1A activation. PACER is an approach that can be widely applied to uncover genetic and environmental determinants of pre-malignant clonal expansion in blood and other tissues.
]]></description>
<dc:creator>Weinstock, J. S.</dc:creator>
<dc:creator>Gopakumar, J.</dc:creator>
<dc:creator>Burugula, B. B.</dc:creator>
<dc:creator>Uddin, M. M.</dc:creator>
<dc:creator>Jahn, N.</dc:creator>
<dc:creator>Belk, J. A.</dc:creator>
<dc:creator>Daniel, B.</dc:creator>
<dc:creator>Ly, N.</dc:creator>
<dc:creator>Mack, T. M.</dc:creator>
<dc:creator>Laurie, C. A.</dc:creator>
<dc:creator>Broome, J. G.</dc:creator>
<dc:creator>Taylor, K. D.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Sinner, M. F.</dc:creator>
<dc:creator>von Falkenhausen, A. S.</dc:creator>
<dc:creator>Kaab, S.</dc:creator>
<dc:creator>Shuldiner, A. R.</dc:creator>
<dc:creator>O'Connell, J. R.</dc:creator>
<dc:creator>Lewis, J. P.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Barnes, K. C.</dc:creator>
<dc:creator>Chami, N.</dc:creator>
<dc:creator>Kenny, E. E.</dc:creator>
<dc:creator>Loos, R. J. F.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Hou, L.</dc:creator>
<dc:creator>Lloyd-Jones, D. M.</dc:creator>
<dc:creator>Redline, S.</dc:creator>
<dc:creator>Cade, B. E.</dc:creator>
<dc:creator>Psaty, B. M.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Brody, J. A.</dc:creator>
<dc:creator>Silverman, E. K.</dc:creator>
<dc:creator>Yun, J. H.</dc:creator>
<dc:creator>Qiao, D.</dc:creator>
<dc:creator>Palmer, N. D.</dc:creator>
<dc:creator>Freedman, B. I.</dc:creator>
<dc:creator>Bowden, D. W.</dc:creator>
<dc:creator>Cho, M. H.</dc:creator>
<dc:creator>DeMeo, D. L.</dc:creator>
<dc:creator>Vasan, R. S.</dc:creator>
<dc:creator>Johnson, A. D.</dc:creator>
<dc:creator>Mat</dc:creator>
<dc:date>2021-12-13</dc:date>
<dc:identifier>doi:10.1101/2021.12.10.471810</dc:identifier>
<dc:title><![CDATA[Clonal hematopoiesis is driven by aberrant activation of TCL1A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.16.472854v1?rss=1">
<title>
<![CDATA[
Fat Body Phospholipid State Dictates Hunger Driven Feeding Behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.16.472854v1?rss=1</link>
<description><![CDATA[
Diet-induced obesity (DIO) leads to dysfunctional feeding behavior. However, the precise molecular nodes underlying diet-induced dysregulation of satiety sensing and feeding motivation are poorly understood. The fruit fly is a simple genetic model system yet displays significant evolutionary conservation to mammalian nutrient sensing and energy balance. Using a longitudinal high sugar regime in Drosophila, we sought to address how lipid alteration in fat cells alters feeding behavior. We find that prolonged exposure to HSD degrades the hunger-driven feeding (HDF) response. Lipidomics analysis reveals that longitudinal exposure to HSD significantly alters whole body phospholipid profiles. By performing a systematic screen for phospholipid enzymes, we identify Pect as a critical regulator of hunger-driven feeding. Pect is a rate-limiting enzyme in the phosphatidylethanolamine (PE) biosynthesis pathway and the fly ortholog of human PCYT2. We show that disrupting Pect only in the fat body causes insulin-resistant phenotypes and a loss of hunger-driven feeding. Excitingly, we find that overexpression of Pect restores HSD-induced loss of hunger-driven feeding response. Strikingly human studies have noted a correlation between PCYT2/Pect levels and clinical obesity. Now, our unbiased studies in Drosophila provide specific genetic evidence for Pect in maintaining nutrient sensing during DIO. Our study provides novel insights into the role of phospholipids in interorgan communication of nutrient status.

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]]></description>
<dc:creator>Kelly, K. P.</dc:creator>
<dc:creator>Alassaf, M.</dc:creator>
<dc:creator>Sullivan, C. E.</dc:creator>
<dc:creator>Brent, A. E.</dc:creator>
<dc:creator>Goldberg, Z. H.</dc:creator>
<dc:creator>Poling, M. E.</dc:creator>
<dc:creator>Dubrulle, J.</dc:creator>
<dc:creator>Rajan, A.</dc:creator>
<dc:date>2021-12-17</dc:date>
<dc:identifier>doi:10.1101/2021.12.16.472854</dc:identifier>
<dc:title><![CDATA[Fat Body Phospholipid State Dictates Hunger Driven Feeding Behavior]]></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/10.1101/2021.12.19.473356v1?rss=1">
<title>
<![CDATA[
Identification and characterization of BrxR as a regulatory gene in the BREX phage restriction system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.19.473356v1?rss=1</link>
<description><![CDATA[
Bacteriophage exclusion ( BREX) phage restriction systems are found in a wide range of bacteria. Various BREX systems encode unique combinations of proteins that usually include a site-specific methyltransferase; none appear to contain a nuclease. Here we describe the identification and characterization of a Type I BREX system from Acinetobacter and the effect of deleting each BREX ORF on growth, methylation, and restriction. We identified a previously uncharacterized gene in the BREX operon that is dispensable for methylation but involved in restriction. Biochemical and crystallographic analyses of this factor, which we term BrxR ( BREX Regulator), demonstrate that it forms a homodimer and specifically binds a DNA target site upstream of its transcription start site. Deletion of the BrxR gene causes cell toxicity, reduces restriction, and significantly increases the expression of BrxC. In contrast, the introduction of a premature stop codon into the BrxR gene, or a point mutation blocking its DNA binding ability, has little effect on restriction, implying that the BrxR coding sequence and BrxR protein play independent functional roles. We speculate that the BrxR coding sequence is involved in cis regulation of anti-phage activity, while the BrxR protein plays an additional regulatory role, perhaps during horizontal transfer.
]]></description>
<dc:creator>Luyten, Y.</dc:creator>
<dc:creator>Hausman, D.</dc:creator>
<dc:creator>Young, J. C.</dc:creator>
<dc:creator>Doyle, L. A.</dc:creator>
<dc:creator>Ubilla-Rodriguez, N. C.</dc:creator>
<dc:creator>Lambert, A. R.</dc:creator>
<dc:creator>Arroyo, C.</dc:creator>
<dc:creator>Forsberg, K. J.</dc:creator>
<dc:creator>Morgan, R. D.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>Kaiser, B. K.</dc:creator>
<dc:date>2021-12-19</dc:date>
<dc:identifier>doi:10.1101/2021.12.19.473356</dc:identifier>
<dc:title><![CDATA[Identification and characterization of BrxR as a regulatory gene in the BREX phage restriction system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.19.473377v1?rss=1">
<title>
<![CDATA[
Simultaneous CUT&Tag profiling of the accessible and silenced regulome in single cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.19.473377v1?rss=1</link>
<description><![CDATA[
Cleavage Under Targets & Tagmentation (CUT&Tag) is an antibody-directed transposase tethering strategy for in situ chromatin profiling in small samples and single cells. We describe a modified CUT&Tag protocol using a mixture of an antibody to the initiation form of RNA Polymerase II (Pol2 Serine-5 phosphate) and an antibody to repressive Polycomb domains (H3K27me3) followed by computational signal deconvolution to produce high-resolution maps of both the active and repressive regulomes in single cells. The ability to seamlessly map active promoters, enhancers and repressive regulatory elements using a single workflow provides a complete regulome profiling strategy suitable for high-throughput single-cell platforms.
]]></description>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Otto, D. J.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2021-12-21</dc:date>
<dc:identifier>doi:10.1101/2021.12.19.473377</dc:identifier>
<dc:title><![CDATA[Simultaneous CUT&Tag profiling of the accessible and silenced regulome in single cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.21.473728v1?rss=1">
<title>
<![CDATA[
Mediator is broadly recruited to gene promoters via a Tail-independent mechanism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.21.473728v1?rss=1</link>
<description><![CDATA[
Mediator (MED) is a conserved factor with important roles in both basal and activated transcription. It is believed that MED plays a direct role in transcriptional regulation at most genes by functionally bridging enhancers and promoters. Here, we investigate the genome-wide roles of yeast MED by rapid depletion of its activator-binding domain (Tail) and monitoring changes in nascent transcription. We find that MED Tail and activator-mediated MED recruitment regulate only a small subset of genes. At most genes, MED bypasses the UAS and is directly recruited to promoters to facilitate transcription initiation. Our results define three classes of genes that differ in PIC assembly pathways and the requirements for MED Tail, SAGA, TFIID and BET factors Bdf1/2. We also find that the depletion of the MED middle module subunit Med7 mimics inactivation of Tail, suggesting a functional link. Our combined results have broad implications for the roles of MED, other coactivators, and mechanisms of transcriptional regulation at different gene classes.
]]></description>
<dc:creator>Warfield, L.</dc:creator>
<dc:creator>Donczew, R.</dc:creator>
<dc:creator>Mahendrawada, L.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:date>2021-12-21</dc:date>
<dc:identifier>doi:10.1101/2021.12.21.473728</dc:identifier>
<dc:title><![CDATA[Mediator is broadly recruited to gene promoters via a Tail-independent mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.22.473781v1?rss=1">
<title>
<![CDATA[
Epistatic selection on a selfish Segregation Distorter supergene: drive, recombination, and genetic load 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.22.473781v1?rss=1</link>
<description><![CDATA[
Meiotic drive supergenes are complexes of alleles at linked loci that together subvert Mendelian segregation to gain preferential transmission. In males, the most common mechanism of drive involves the disruption of sperm bearing alternative alleles. While at least two loci are important for male drive-- the driver and the target-- linked modifiers can enhance drive, creating selection pressure to suppress recombination. In this work, we investigate the evolution and genomic consequences of an autosomal multilocus, male meiotic drive system, Segregation Distorter (SD) in the fruit fly, Drosophila melanogaster. In African populations, the predominant SD chromosome variant, SD-Mal, is characterized by two overlapping, paracentric inversion on chromosome arm 2R and nearly perfect (~100%) transmission. We study the SD-Mal system in detail, exploring its components, chromosomal structure, and evolutionary history. Our findings reveal a recent chromosome-scale selective sweep mediated by strong epistatic selection for haplotypes carrying Sd, the main driving allele, and one or more factors within the double inversion. While most SD-Mal chromosomes are homozygous lethal, SD-Mal haplotypes can recombine with other, complementing haplotypes via crossing over and with wildtype chromosomes only via gene conversion. SD-Mal chromosomes have nevertheless accumulated lethal mutations, excess non-synonymous mutations, and excess transposable element insertions. Therefore, SD-Mal haplotypes evolve as a small, semi-isolated subpopulation with a history of strong selection. These results may explain the evolutionary turnover of SD haplotypes in different populations around the world and have implications for supergene evolution broadly.
]]></description>
<dc:creator>Navarro-Dominguez, B.</dc:creator>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Brand, C.</dc:creator>
<dc:creator>Muirhead, C.</dc:creator>
<dc:creator>Presgraves, D.</dc:creator>
<dc:creator>Larracuente, A. M.</dc:creator>
<dc:date>2021-12-24</dc:date>
<dc:identifier>doi:10.1101/2021.12.22.473781</dc:identifier>
<dc:title><![CDATA[Epistatic selection on a selfish Segregation Distorter supergene: drive, recombination, and genetic load]]></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/10.1101/2021.12.23.474064v1?rss=1">
<title>
<![CDATA[
ROKET: Associating Somatic Mutation with Clinical Outcomes through Kernel Regression and Optimal Transport 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.23.474064v1?rss=1</link>
<description><![CDATA[
Somatic mutations in cancer patients are inherently sparse and potentially high dimensional. Cancer patients may share the same set of deregulated biological processes perturbed by different sets of somatically mutated genes. Therefore, when assessing the associations between somatic mutations and clinical outcomes, gene-by-gene analyses is often under-powered because it does not capture the complex disease mechanisms shared across cancer patients. Rather than testing genes one by one, an intuitive approach is to aggregate somatic mutation data of multiple genes to assess the joint association. The challenge is how to aggregate such information. Building on the optimal transport method, we propose a principled approach to estimate the similarity of somatic mutation profiles of multiple genes between tumor samples, while accounting for gene-gene similarity defined by gene annotations or empirical mutational patterns. Using such similarities, we can assess the associations between somatic mutations and clinical outcomes by kernel regression. We have applied our method to analyze somatic mutation data of 17 cancer types and identified at least three cancer types harboring associations between somatic mutations and overall survival, progression-free interval or cytolytic activity.
]]></description>
<dc:creator>Little, P.</dc:creator>
<dc:creator>Hsu, L.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2021-12-24</dc:date>
<dc:identifier>doi:10.1101/2021.12.23.474064</dc:identifier>
<dc:title><![CDATA[ROKET: Associating Somatic Mutation with Clinical Outcomes through Kernel Regression and Optimal Transport]]></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/10.1101/2022.01.06.475296v1?rss=1">
<title>
<![CDATA[
Translational buffering by ribosome stalling in upstream open reading frames 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.06.475296v1?rss=1</link>
<description><![CDATA[
Upstream open reading frames (uORFs) are present in over half of all human mRNAs. uORFs can potently regulate the translation of downstream open reading frames by several mechanisms: siphoning away scanning ribo-somes, regulating re-initiation, and allowing interactions between scanning and elongating ribosomes. However, the consequences of these different mechanisms for the regulation of protein expression remain incompletely understood. Here, we performed systematic measurements on the uORF-containing 5 UTR of the cytomegaloviral UL4 mRNA to test alternative models of uORF-mediated regulation in human cells. We find that a terminal diproline-dependent elongating ribosome stall in the UL4 uORF prevents decreases in main ORF translation when ribosome loading onto the mRNA is reduced. This uORF-mediated buffering is insensitive to the location of the ribosome stall along the uORF. Computational kinetic modeling based on our measurements suggests that scanning ribosomes dissociate rather than queue when they collide with stalled elongating ribosomes within the UL4 uORF. We identify several human uORFs that repress main ORF translation via a similar terminal diproline motif. We propose that ribosome stalls in uORFs provide a general mechanism for buffering against reductions in main ORF translation during stress and developmental transitions.
]]></description>
<dc:creator>Bottorff, T. A.</dc:creator>
<dc:creator>Geballe, A. P.</dc:creator>
<dc:creator>Subramaniam, A. R.</dc:creator>
<dc:date>2022-01-07</dc:date>
<dc:identifier>doi:10.1101/2022.01.06.475296</dc:identifier>
<dc:title><![CDATA[Translational buffering by ribosome stalling in upstream open reading frames]]></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/10.1101/2022.01.07.475392v1?rss=1">
<title>
<![CDATA[
Oncogenic role of a developmentally regulated NTRK2 splice variant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.07.475392v1?rss=1</link>
<description><![CDATA[
Temporally-regulated alternative splicing choices are vital for proper development yet the wrong splice choice may be detrimental. Here we highlight a novel role for the neurotrophin receptor splice variant TrkB.T1 in neurodevelopment, embryogenesis, transformation, and oncogenesis across multiple tumor types in both humans and mice. TrkB.T1 is the predominant NTRK2 isoform across embryonic organogenesis and forced over-expression of this embryonic pattern causes multiple solid and nonsolid tumors in mice in the context of tumor suppressor loss. TrkB.T1 also emerges the predominant NTRK isoform expressed in a wide range of adult and pediatric tumors, including those harboring TRK fusions. Affinity purification-mass spectrometry (AP-MS) proteomic analysis reveals TrkB.T1 has distinct interactors with known developmental and oncogenic signaling pathways such as Wnt, TGF-{beta}, Hedgehog, and Ras. From alterations in splicing factors to changes in gene expression, the discovery of isoform specific oncogenes with embryonic ancestry has the potential to shape the way we think about developmental systems and oncology.
]]></description>
<dc:creator>Pattwell, S. S.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Nuechterlein, N.</dc:creator>
<dc:creator>Zager, M.</dc:creator>
<dc:creator>Loeb, K. R.</dc:creator>
<dc:creator>Cimino, P. J.</dc:creator>
<dc:creator>Holland, N. C.</dc:creator>
<dc:creator>Reche-Ley, N.</dc:creator>
<dc:creator>Bolouri, H.</dc:creator>
<dc:creator>Bonin, D. A. A.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:creator>Phadnis, V. V.</dc:creator>
<dc:creator>Ozawa, T.</dc:creator>
<dc:creator>Wagner, M. J.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:date>2022-01-09</dc:date>
<dc:identifier>doi:10.1101/2022.01.07.475392</dc:identifier>
<dc:title><![CDATA[Oncogenic role of a developmentally regulated NTRK2 splice variant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.09.475586v1?rss=1">
<title>
<![CDATA[
Patterns of Structural Variation Define Prostate Cancer Across Disease States 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.09.475586v1?rss=1</link>
<description><![CDATA[
The complex genomic landscape of prostate cancer evolves across disease states under therapeutic pressure directed toward inhibiting androgen receptor (AR) signaling. While significantly altered genes in prostate cancer have been extensively defined, there have been fewer systematic analyses of how structural variation shapes the genomic landscape of this disease across disease states. We uniformly characterized structural alterations across 278 localized and 143 metastatic prostate cancers profiled by whole genome and transcriptome sequencing. We observed distinct significantly recurrent breakpoints in localized and metastatic castration-resistant prostate cancers (mCRPC), with pervasive alterations in noncoding regions flanking the AR, MYC, FOXA1, and LSAMP genes enriched in mCRPC and TMPRSS2-ERG rearrangements enriched in localized prostate cancer. We defined nine subclasses of mCRPC based on signatures of structural variation, each associated with distinct genetic features and clinical outcomes. Our results comprehensively define patterns of structural variation in prostate cancer and identify clinically actionable subgroups based on whole genome profiling.
]]></description>
<dc:creator>Zhou, M.</dc:creator>
<dc:creator>Ko, M.</dc:creator>
<dc:creator>Hoge, A. C.</dc:creator>
<dc:creator>Luu, K.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Russell, M. L.</dc:creator>
<dc:creator>Hannon, W. W.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Carrot-Zhang, J.</dc:creator>
<dc:creator>Beroukhim, R.</dc:creator>
<dc:creator>Van Allen, E.</dc:creator>
<dc:creator>Choudhury, A. D.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Freedman, M. L.</dc:creator>
<dc:creator>Taplin, M.-E.</dc:creator>
<dc:creator>Meyerson, M.</dc:creator>
<dc:creator>Viswanathan, S. R.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:date>2022-01-11</dc:date>
<dc:identifier>doi:10.1101/2022.01.09.475586</dc:identifier>
<dc:title><![CDATA[Patterns of Structural Variation Define Prostate Cancer Across Disease States]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.12.476097v1?rss=1">
<title>
<![CDATA[
Myeloid-biased HSC require Semaphorin 4A from the bone marrow niche for self-renewal under stress and life-long persistence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.12.476097v1?rss=1</link>
<description><![CDATA[
Tissue stem cells are hierarchically organized. Those that are most primitive serve as key drivers of regenerative response but the signals that selectively preserve their functional integrity are largely unknown. Here, we identify a secreted factor, Semaphorin 4A (Sema4A), as a specific regulator of myeloid-biased hematopoietic stem cells (myHSC), which are positioned at the top of the HSC hierarchy. Lack of Sema4A leads to exaggerated myHSC (but not downstream "balanced" HSC) proliferation after acute inflammatory stress, indicating that Sema4A enforces myHSC quiescence. Strikingly, aged Sema4A knock-out myHSC expand but almost completely lose reconstitution capacity. The effect of Sema4A is non cell-autonomous, since upon transplantation into Sema4A-deficient environment, wild-type myHSC excessively proliferate but fail to engraft long-term. Sema4A constrains inflammatory signaling in myHSC and acts via a surface receptor Plexin-D1. Our data support a model whereby the most primitive tissue stem cells critically rely on a dedicated signal from the niche for self-renewal and life-long persistence.
]]></description>
<dc:creator>Toghani, D.</dc:creator>
<dc:creator>Zeng, S.</dc:creator>
<dc:creator>Mahammadov, E.</dc:creator>
<dc:creator>Crosse, E. I.</dc:creator>
<dc:creator>Seyedhassantehrani, N.</dc:creator>
<dc:creator>Burns, C.</dc:creator>
<dc:creator>Gravano, D.</dc:creator>
<dc:creator>Radtke, S.</dc:creator>
<dc:creator>Kiem, H.-P.</dc:creator>
<dc:creator>Rodriguez, S.</dc:creator>
<dc:creator>Carlesso, N.</dc:creator>
<dc:creator>Pradeep, A.</dc:creator>
<dc:creator>Wilson, N. K.</dc:creator>
<dc:creator>Kinston, S. J.</dc:creator>
<dc:creator>Gottgens, B.</dc:creator>
<dc:creator>Nerlov, C.</dc:creator>
<dc:creator>Pietras, E.</dc:creator>
<dc:creator>Mesnieres, M.</dc:creator>
<dc:creator>Maes, C.</dc:creator>
<dc:creator>Kumanogoh, A.</dc:creator>
<dc:creator>Worzfeld, T.</dc:creator>
<dc:creator>Kharchenko, P.</dc:creator>
<dc:creator>Scialdone, A.</dc:creator>
<dc:creator>Spencer, J. A.</dc:creator>
<dc:creator>Silberstein, L.</dc:creator>
<dc:date>2022-01-12</dc:date>
<dc:identifier>doi:10.1101/2022.01.12.476097</dc:identifier>
<dc:title><![CDATA[Myeloid-biased HSC require Semaphorin 4A from the bone marrow niche for self-renewal under stress and life-long persistence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.15.476465v1?rss=1">
<title>
<![CDATA[
A giant virus genome is densely packaged by stable nucleosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.15.476465v1?rss=1</link>
<description><![CDATA[
The two doublet histones of Marseillevirus are distantly related to the four eukaryotic core histones and wrap 121 basepairs of DNA to form remarkably similar nucleosomes. By permeabilizing Marseillevirus virions and performing genome-wide nuclease digestion, chemical cleavage and mass spectrometry assays, we find that the higher-order organization of Marseillevirus chromatin fundamentally differs from that of eukaryotes. Marseillevirus nucleosomes fully protect DNA within virions as closely abutted 121-bp DNA wrapped cores without linker DNA or phasing along genes. Likewise, we observed that a large fraction of the nucleosomes reconstituted onto multi-copy tandem repeats of a nucleosome positioning sequence are tightly packed. Dense promiscuous packing of fully wrapped nucleosomes rather than "beads-on-a-string" with genic punctuation represents a new mode of DNA packaging by histones. We suggest that doublet histones have evolved for viral genome protection and may resemble an early stage of histone differentiation leading to the eukaryotic octameric nucleosome.
]]></description>
<dc:creator>Bryson, T. D.</dc:creator>
<dc:creator>De Ioannes, P.</dc:creator>
<dc:creator>Valencia-Sanchez, M. I.</dc:creator>
<dc:creator>Henikoff, J. G.</dc:creator>
<dc:creator>Talbert, P. B.</dc:creator>
<dc:creator>La Scola, B.</dc:creator>
<dc:creator>Armache, K.-J.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2022-01-16</dc:date>
<dc:identifier>doi:10.1101/2022.01.15.476465</dc:identifier>
<dc:title><![CDATA[A giant virus genome is densely packaged by stable nucleosomes]]></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/10.1101/2022.01.19.475968v1?rss=1">
<title>
<![CDATA[
Single-Cell Transcriptomic Analyses of Tumor Ecosystems and Spatial Architectures in Human Small Cell Lung Cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.19.475968v1?rss=1</link>
<description><![CDATA[
Small cell lung cancer (SCLC) is a highly heterogenous disease characterized by aggressive phenotypes and poor prognosis. In order to dissect the cellular components and their spatial distribution in SCLC ecosystems, here we performed single-cell RNA sequencing and spatial transcriptomics analyses of 12 resected human primary SCLCs. Our analysis revealed extensive heterogeneity of both tumor and non-tumor cells, and a predominant immunosuppressive microenvironment. Importantly, multiple hybrid tumor cell states were unmasked, including hybrid SCLC and lung adenocarcinoma cells as well as hybrid tumor and immune cells, indicating high phenotypic plasticity of tumor cells. Moreover, we revealed the cellular origin and evolution of these hybrid tumor cells, and critical roles of tumor-stromal cellular crosstalk in driving the development of hybrid cells. In conclusion, this study deepens our understanding of human SCLC ecosystems and provides rationales to target the hybrid tumor cells and immunosuppressive microenvironment for SCLC therapies.
]]></description>
<dc:creator>Ouyang, H.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>He, C.</dc:creator>
<dc:creator>Sheng, L.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Pan, D.</dc:creator>
<dc:creator>Augert, A.</dc:creator>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Jia, D.</dc:creator>
<dc:date>2022-01-21</dc:date>
<dc:identifier>doi:10.1101/2022.01.19.475968</dc:identifier>
<dc:title><![CDATA[Single-Cell Transcriptomic Analyses of Tumor Ecosystems and Spatial Architectures in Human Small Cell Lung Cancers]]></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/10.1101/2022.01.30.478369v1?rss=1">
<title>
<![CDATA[
mTOR-regulated Mitochondrial Metabolism Limits Mycobacterium-induced Cytotoxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.30.478369v1?rss=1</link>
<description><![CDATA[
Necrosis of macrophages in the tuberculous granuloma represents a major pathogenic event in tuberculosis. Through a zebrafish forward genetic screen, we identified the mTOR kinase, a master regulator of metabolism, as an early host resistance factor in tuberculosis. We found that mTOR complex 1 protects macrophages from mycobacterium-induced death by enabling infection-induced increases in mitochondrial energy metabolism fueled by glycolysis. These metabolic adaptations are required to prevent mitochondrial damage and death caused by the secreted mycobacterial virulence determinant ESAT-6. Thus, the host can effectively counter this early critical mycobacterial virulence mechanism simply by regulating energy metabolism, thereby allowing pathogen-specific immune mechanisms time to develop. Our findings may explain why Mycobacterium tuberculosis, albeit humanitys most lethal pathogen, is successful in only a minority of infected individuals.
]]></description>
<dc:creator>Pagan, A. J.</dc:creator>
<dc:creator>Lee, L. J.</dc:creator>
<dc:creator>Edwards-Hicks, J.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:creator>Tobin, D. M.</dc:creator>
<dc:creator>Busch-Nentwich, E. M.</dc:creator>
<dc:creator>Pearce, E. L.</dc:creator>
<dc:creator>Ramakrishnan, L.</dc:creator>
<dc:date>2022-01-30</dc:date>
<dc:identifier>doi:10.1101/2022.01.30.478369</dc:identifier>
<dc:title><![CDATA[mTOR-regulated Mitochondrial Metabolism Limits Mycobacterium-induced Cytotoxicity]]></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/10.1101/2022.01.31.478548v1?rss=1">
<title>
<![CDATA[
HOXD13 is a direct EWS-FLI1 target and moderates fusion-dependent transcriptional states 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.31.478548v1?rss=1</link>
<description><![CDATA[
Oncogenic fusion proteins display exquisite tissue specificity, revealing that malignant transformation requires cooperation with cell-autonomous factors. Recent studies have also demonstrated that tumorigenicity of Ewing sarcoma requires precise regulation of the transcriptional activity of the EWS-FLI1 oncogenic driver. Here we show that the developmentally and anatomically restricted transcription factor HOXD13 is a direct target of EWS-FLI1. Transcriptomic and CUT&RUN studies revealed that HOXD13 binds active, fusion-bound enhancers, resulting in altered expression of EWS-FLI1-induced targets. More strikingly, HOXD13 was found to bind and activate cis-regulatory regions of genes that are normally repressed by EWS-FLI1. Single-cell sequencing demonstrated marked intra-tumoral heterogeneity of HOXD13 transcriptional activity and revealed that antagonism between HOXD13-mediated gene activation and EWS-FLI1-dependent gene repression confers a spectrum of transcriptional cell states along a mesenchymal axis. Thus, HOXD13 serves as an internal rheostat for EWS-FLI1 activity, providing a paradigm for tissue-specific transcription factors as critical partners in fusion-driven cancers.
]]></description>
<dc:creator>Apfelbaum, A. A.</dc:creator>
<dc:creator>Wu, F.</dc:creator>
<dc:creator>Hawkins, A.</dc:creator>
<dc:creator>Jimenez, J. A.</dc:creator>
<dc:creator>Taylor, S. D.</dc:creator>
<dc:creator>Pfaltzgraff, E. R.</dc:creator>
<dc:creator>Song, J. Y.</dc:creator>
<dc:creator>Hall, C.</dc:creator>
<dc:creator>Wellik, D. M.</dc:creator>
<dc:creator>Ljungman, M.</dc:creator>
<dc:creator>Furlan, S. N.</dc:creator>
<dc:creator>Ryan, R. J. H.</dc:creator>
<dc:creator>Sarthy, J. F.</dc:creator>
<dc:creator>Lawlor, E. R.</dc:creator>
<dc:date>2022-02-01</dc:date>
<dc:identifier>doi:10.1101/2022.01.31.478548</dc:identifier>
<dc:title><![CDATA[HOXD13 is a direct EWS-FLI1 target and moderates fusion-dependent transcriptional states]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.03.478946v1?rss=1">
<title>
<![CDATA[
Antibody affinity and concentration of convalescent sera provide context for reduced SARS-CoV-2 Omicron spike affinity of therapeutic antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.03.478946v1?rss=1</link>
<description><![CDATA[
We assessed the affinities of the therapeutic monoclonal antibodies (mAbs) cilgavimab, tixagevimab, sotrovimab, casirivimab, and imdevimab to the receptor binding domain (RBD) of wild type, Delta, and Omicron spike. The Omicron RBD affinities of cilgavimab, tixagevimab, casirivimab, and imdevimab decreased by at least two orders of magnitude relative to their wild type equivalents, whereas sotrovimab binding was less severely impacted. These affinity reductions correlate with reduced antiviral activities of these antibodies, suggesting that simple affinity measurements can serve as an indicator for activity before challenging and time-consuming virus neutralization assays are performed. We also compared the properties of these antibodies to serological fingerprints (affinities and concentrations) of wild type RBD specific antibodies in 74 convalescent sera. The affinities of the therapeutic mAbs to wild type and Delta RBD were in the same range as the polyclonal response in the convalescent sera indicative of their high antiviral activities against these variants. However, for Omicron RBD, only sotrovimab retained affinities that were within the range of the polyclonal response, in agreement with its high activity against Omicron. Serological fingerprints thus provide important context to affinities and antiviral activity of mAb drugs and could guide the development of new therapeutics.
]]></description>
<dc:creator>Fiedler, S.</dc:creator>
<dc:creator>Devenish, S. R. A.</dc:creator>
<dc:creator>Morgunov, A. S.</dc:creator>
<dc:creator>Ilsley, A.</dc:creator>
<dc:creator>Ricci, F.</dc:creator>
<dc:creator>Emmenegger, M.</dc:creator>
<dc:creator>Kosmoliaptsis, V.</dc:creator>
<dc:creator>Theel, E. S.</dc:creator>
<dc:creator>Mills, J. R.</dc:creator>
<dc:creator>Sholukh, A. M.</dc:creator>
<dc:creator>Aguzzi, A. A. A.</dc:creator>
<dc:creator>Iwasaki, A.</dc:creator>
<dc:creator>Lynn, A. K.</dc:creator>
<dc:creator>Knowles, T. P. J.</dc:creator>
<dc:date>2022-02-04</dc:date>
<dc:identifier>doi:10.1101/2022.02.03.478946</dc:identifier>
<dc:title><![CDATA[Antibody affinity and concentration of convalescent sera provide context for reduced SARS-CoV-2 Omicron spike affinity of therapeutic antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.07.479443v1?rss=1">
<title>
<![CDATA[
Olverembatinib inhibits SARS-CoV-2-Omicron variant-mediated cytokine release 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.07.479443v1?rss=1</link>
<description><![CDATA[
The Omicron variant has become dominant in the U.S. and around the world. This variant is found to be 2-fold more infectious than the Delta variant, posing a significant threat of severe cases and death. We and others have recently reported that the N-terminus domain (NTD) of the SARS-CoV-2 of various variants is responsible for inducing cytokine release in human PBMCs. Here, we demonstrate that the NTD of the Omicron variant remains highly effective at inducing cytokine release in human PBMCs. Furthermore, we show that Ponatinib and a novel compound, Olverembatinib, are potent Omicron NTD-mediated cytokine release inhibitors. Target profiling revealed that Olverembatinib blocks most of the previously identified kinases responsible for cytokine release. Together, we propose that Ponatinib and Olverembatinib may represent an attractive therapeutic option for treating moderate to severe COVID-19 cases.

HIGHLIGHTSO_LIThe N-terminus domain (NTD) of the SARS-CoV-2 Omicron variant strongly induces multiple inflammatory molecules in PBMCs, unaffected by the mutations observed in the NTD.
C_LIO_LIThe cytokine release mediated by the Omicron variant is comparable to the Delta variant.
C_LIO_LIOlverembatinib, a clinical-stage multi-kinase inhibitor, potently inhibits Omicron NTD-mediated cytokine release.
C_LIO_LIOlverembatinib could relieve severe symptoms associated with COVID-19 Omicron and Delta variants.
C_LI
]]></description>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Holland, E.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:date>2022-02-09</dc:date>
<dc:identifier>doi:10.1101/2022.02.07.479443</dc:identifier>
<dc:title><![CDATA[Olverembatinib inhibits SARS-CoV-2-Omicron variant-mediated cytokine release]]></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/10.1101/2022.02.09.479546v1?rss=1">
<title>
<![CDATA[
Narrow transmission bottlenecks and limited within-host viral diversity during a SARS-CoV-2 outbreak on a fishing boat 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.09.479546v1?rss=1</link>
<description><![CDATA[
The long-term evolution of viruses is ultimately due to viral mutants that arise within infected individuals and transmit to other individuals. Here we use deep sequencing to investigate the transmission of viral genetic variation among individuals during a SARS-CoV-2 outbreak that infected the vast majority of crew members on a fishing boat. We deep-sequenced nasal swabs to characterize the within-host viral population of infected crew members, using experimental duplicates and strict computational filters to ensure accurate variant calling. We find that within-host viral diversity is low in infected crew members. The mutations that did fix in some crew members during the outbreak are not observed at detectable frequencies in any of the sampled crew members in which they are not fixed, suggesting viral evolution involves occasional fixation of low-frequency mutations during transmission rather than persistent maintenance of within-host viral diversity. Overall, our results show that strong transmission bottlenecks dominate viral evolution even during a superspreading event with a very high attack rate.
]]></description>
<dc:creator>Hannon, W. H.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Shrestha, L.</dc:creator>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2022-02-09</dc:date>
<dc:identifier>doi:10.1101/2022.02.09.479546</dc:identifier>
<dc:title><![CDATA[Narrow transmission bottlenecks and limited within-host viral diversity during a SARS-CoV-2 outbreak on a fishing boat]]></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/10.1101/2022.02.17.480914v1?rss=1">
<title>
<![CDATA[
Immunization with a self-assembling nanoparticle vaccine displaying EBV gH/gL protects humanized mice against lethal viral challenge 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.17.480914v1?rss=1</link>
<description><![CDATA[
Summary/AbstractEpstein-Barr virus (EBV) is a cancer-associated pathogen responsible for 140,000 deaths per year. EBV is also the etiological agent of infectious mononucleosis and is associated with multiple sclerosis and rheumatoid arthritis. Thus, an EBV vaccine could alleviate significant morbidity and mortality. EBV is orally transmitted and has tropism for both epithelial cells and B cells which are present in the oral cavity. Therefore, a prophylactic vaccine would need to prevent infection of both cell types. Passive transfer neutralizing monoclonal antibodies targeting the viral gH/gL glycoprotein complex prevent experimental EBV infection in humanized mice and rhesus macaques, suggesting that gH/gL is an attractive vaccine candidate. Here, we produced and evaluated the immunogenicity of several nanoparticle immunogens displaying gH/gL with distinct valencies and geometries. After one or two immunizations, all nanoparticles elicited superior binding and neutralizing titers relative to monomeric gH/gL. Antibodies elicited by a computationally designed self-assembling nanoparticle that displays 60 copies of the gH/gL protein conferred protection against a lethal dose of EBV in a humanized mouse challenge model, whereas antibodies elicited by monomeric gH/gL did not. Taken together, these data motivate further development of gH/gL nanoparticle vaccine candidates for EBV.
]]></description>
<dc:creator>Malhi, H.</dc:creator>
<dc:creator>Homad, L. J.</dc:creator>
<dc:creator>Wan, Y.-H.</dc:creator>
<dc:creator>Poudel, B.</dc:creator>
<dc:creator>Fiala, B.</dc:creator>
<dc:creator>Borst, A. J.</dc:creator>
<dc:creator>Wang, J. Y.</dc:creator>
<dc:creator>Walkey, C.</dc:creator>
<dc:creator>Price, J.</dc:creator>
<dc:creator>Wall, A.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Moodie, Z.</dc:creator>
<dc:creator>Handa, S.</dc:creator>
<dc:creator>Correnti, C.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>Olson, J.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:creator>McGuire, A.</dc:creator>
<dc:date>2022-02-19</dc:date>
<dc:identifier>doi:10.1101/2022.02.17.480914</dc:identifier>
<dc:title><![CDATA[Immunization with a self-assembling nanoparticle vaccine displaying EBV gH/gL protects humanized mice against lethal viral challenge]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.24.481899v1?rss=1">
<title>
<![CDATA[
Shifting mutational constraints in the SARS-CoV-2 receptor-binding domain during viral evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.24.481899v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 has evolved variants with substitutions in the spike receptor-binding domain (RBD) that impact its affinity for ACE2 receptor and recognition by antibodies. These substitutions could also shape future evolution by modulating the effects of mutations at other sites--a phenomenon called epistasis. To investigate this possibility, we performed deep mutational scans to measure the effects on ACE2 binding of all single amino-acid mutations in the Wuhan-Hu-1, Alpha, Beta, Delta, and Eta variant RBDs. Some substitutions, most prominently N501Y, cause epistatic shifts in the effects of mutations at other sites, thereby shaping subsequent evolutionary change. These epistatic shifts occur despite high conservation of the overall RBD structure. Our data shed light on RBD sequence-function relationships and facilitate interpretation of ongoing SARS-CoV-2 evolution.
]]></description>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Hannon, W. W.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Hauser, K.</dc:creator>
<dc:creator>Dillen, J. R.</dc:creator>
<dc:creator>Ferri, E.</dc:creator>
<dc:creator>Ghez Farrell, A.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>McCallum, M.</dc:creator>
<dc:creator>Matreyek, K. A.</dc:creator>
<dc:creator>Corti, D.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Snell, G.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2022-02-26</dc:date>
<dc:identifier>doi:10.1101/2022.02.24.481899</dc:identifier>
<dc:title><![CDATA[Shifting mutational constraints in the SARS-CoV-2 receptor-binding domain during viral evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.02.482683v1?rss=1">
<title>
<![CDATA[
Distinct gene expression by expanded clones of quiescent memory CD4+ T cells harboring intact latent HIV-1 proviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.02.482683v1?rss=1</link>
<description><![CDATA[
Antiretroviral therapy controls but does not cure HIV-1 infection due to a reservoir of rare CD4+ T cells harboring latent proviruses. Little is known about the transcriptional program of latent cells. Here we report a novel strategy to enrich clones of latent cells carrying intact, replication- competent HIV-1 proviruses from blood based on their expression of unique T cell receptors. Latent cell enrichment enabled single cell transcriptomic analysis of 1,050 CD4+ T cells belonging to expanded clones harboring intact HIV-1 proviruses from 6 different individuals. The analysis revealed that most of these cells are T effector memory cells that are enriched for expression of HLA-DR, HLA-DP, CD74, CCL5, Granzymes A and K, cystatin F, LYAR and DUSP2. We conclude that expanded clones of latent cells carrying intact HIV-1 proviruses persist preferentially in a distinct CD4+ T cell population opening new possibilities for eradication.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=146 SRC="FIGDIR/small/482683v2_ufig1.gif" ALT="Figure 1">
View larger version (35K):
org.highwire.dtl.DTLVardef@19e89d2org.highwire.dtl.DTLVardef@16a5685org.highwire.dtl.DTLVardef@563a97org.highwire.dtl.DTLVardef@120a224_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Weymar, G. H.</dc:creator>
<dc:creator>Bar-On, Y.</dc:creator>
<dc:creator>Oliveira, T. Y.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Breton, G.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Cohn, L. B.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2022-03-02</dc:date>
<dc:identifier>doi:10.1101/2022.03.02.482683</dc:identifier>
<dc:title><![CDATA[Distinct gene expression by expanded clones of quiescent memory CD4+ T cells harboring intact latent HIV-1 proviruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.02.482711v1?rss=1">
<title>
<![CDATA[
Defining cellular population dynamics at single cell resolution during prostate cancer progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.02.482711v1?rss=1</link>
<description><![CDATA[
Advanced prostate malignancies are a leading cause of cancer-related deaths in men, in large part due to our incomplete understanding of cellular drivers of disease progression. We investigated prostate cancer cell dynamics at single-cell resolution from disease onset to the development of androgen independence in vivo. We observe a dramatic expansion of a castration-resistant intermediate luminal cell type that correlates with treatment resistance and poor prognosis in human patients. Moreover, transformed epithelial cells and associated fibroblasts create a microenvironment conducive to pro-tumorigenic immune infiltration, which is in part androgen responsive. Androgen independent prostate cancer leads to significant diversification of intermediate luminal cell populations characterized by a range of androgen signaling activity inversely correlated with proliferation and mRNA translation. Accordingly, distinct epithelial populations are exquisitely sensitive to translation inhibition which leads to epithelial cell death, loss of pro-tumorigenic signaling, and decreased tumor heterogeneity. Our findings reveal a complex tumor environment largely dominated by castration-resistant luminal cells and immunosuppressive infiltrates.
]]></description>
<dc:creator>Germanos, A. A.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Goddard, E. T.</dc:creator>
<dc:creator>Coleman, I. M.</dc:creator>
<dc:creator>Ku, A. T.</dc:creator>
<dc:creator>Wilkinson, S.</dc:creator>
<dc:creator>Amezquita, R. A.</dc:creator>
<dc:creator>Zager, M.</dc:creator>
<dc:creator>Long, A.</dc:creator>
<dc:creator>Yang, Y. C.</dc:creator>
<dc:creator>Bielas, J. H.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Ghajar, C. M.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Sowalsky, A. G.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:creator>Hsieh, A. C.</dc:creator>
<dc:date>2022-03-04</dc:date>
<dc:identifier>doi:10.1101/2022.03.02.482711</dc:identifier>
<dc:title><![CDATA[Defining cellular population dynamics at single cell resolution during prostate cancer progression]]></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/10.1101/2022.03.09.483595v1?rss=1">
<title>
<![CDATA[
Activation of targetable inflammatory immune signaling is seen in Myelodysplastic Syndromes with SF3B1 mutations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.09.483595v1?rss=1</link>
<description><![CDATA[
BackgroundMutations in the SF3B1 splicing factor are commonly seen in Myelodysplastic syndromes (MDS) and Acute Myeloid Leukemia (AML), yet the specific oncogenic pathways activated by missplicing have not been fully elucidated. Inflammatory immune pathways have been shown to play roles in pathogenesis of MDS, though the exact mechanisms of their activation in splicing mutant cases are not well understood.

MethodsRNA-seq data from SF3B1 mutant samples was analyzed and functional roles of IRAK4 isoforms were determined. Efficacy of IRAK4 inhibition was evaluated in pre-clinical models of MDS/AML

ResultsRNA-seq splicing analysis of innate immune mediators in SF3B1 mutant MDS samples revealed retention of full-length exon 6 of interleukin-1 receptor-associated kinase 4 (IRAK4), a critical downstream mediator that links the Myddosome to inflammatory NF-kB activation. Exon 6 retention leads to a longer isoform, encoding a protein (IRAK4-Long) that contains the entire death domain and kinase domain, leading to maximal activation of NF-kB. Cells with wild-type SF3B1 contain smaller IRAK4 isoforms that are targeted for proteosomal degradation. Expression of IRAK4-Long in SF3B1 mutant cells induces TRAF6 activation leading to K63-linked ubiquitination of CDK2, associated with a block in hematopoietic differentiation. Inhibition of IRAK4 with CA-4948, leads to reduction in NF-kB activation, inflammatory cytokine production, enhanced myeloid differentiation in vitro and reduced leukemic growth in xenograft models.

ConclusionsSF3B1 mutation leads to expression of a therapeutically targetable, longer, oncogenic IRAK4 isoform in AML/MDS models.
]]></description>
<dc:creator>Choudhary, G.</dc:creator>
<dc:creator>Pellagatti, A.</dc:creator>
<dc:creator>Agianian, B.</dc:creator>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Bhagat, T. D.</dc:creator>
<dc:creator>Gordon-Mitchell, S.</dc:creator>
<dc:creator>Pandey, S.</dc:creator>
<dc:creator>Shah, N.</dc:creator>
<dc:creator>Aluri, S.</dc:creator>
<dc:creator>Schwartz, L.</dc:creator>
<dc:creator>Steeples, V.</dc:creator>
<dc:creator>Booher, R.</dc:creator>
<dc:creator>Ramachandra, M.</dc:creator>
<dc:creator>Samson, M.</dc:creator>
<dc:creator>Carbajal, M.</dc:creator>
<dc:creator>Pradhan, K.</dc:creator>
<dc:creator>Bowman, T.</dc:creator>
<dc:creator>Pillai, M. M.</dc:creator>
<dc:creator>Will, B.</dc:creator>
<dc:creator>Wickrema, A.</dc:creator>
<dc:creator>Shastri, A.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:creator>Martell, R. E.</dc:creator>
<dc:creator>Steidl, U.</dc:creator>
<dc:creator>Gavathiotis, E.</dc:creator>
<dc:creator>Boultwood, J.</dc:creator>
<dc:creator>Starczynowski, D. T.</dc:creator>
<dc:creator>Verma, A.</dc:creator>
<dc:date>2022-03-09</dc:date>
<dc:identifier>doi:10.1101/2022.03.09.483595</dc:identifier>
<dc:title><![CDATA[Activation of targetable inflammatory immune signaling is seen in Myelodysplastic Syndromes with SF3B1 mutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.12.484088v1?rss=1">
<title>
<![CDATA[
The SARS-CoV-2 Delta variant induces an antibody response largely focused on class 1 and 2 antibody epitopes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.12.484088v1?rss=1</link>
<description><![CDATA[
Exposure histories to SARS-CoV-2 variants and vaccinations will shape the specificity of antibody responses. To understand the specificity of Delta-elicited antibody immunity, we characterize the polyclonal antibody response elicited by primary or mRNA vaccine-breakthrough Delta infections. Both types of infection elicit a neutralizing antibody response focused heavily on the receptor-binding domain (RBD). We use deep mutational scanning to show that mutations to the RBDs class 1 and class 2 epitopes, including sites 417, 478, and 484-486 often reduce binding of these Delta-elicited antibodies. The anti-Delta antibody response is more similar to that elicited by early 2020 viruses than the Beta variant, with mutations to the class 1 and 2, but not class 3 epitopes, having the largest effects on polyclonal antibody binding. In addition, mutations to the class 1 epitope (e.g., K417N) tend to have larger effects on antibody binding and neutralization in the Delta spike than in the D614G spike, both for vaccine- and Delta-infection-elicited antibodies. These results help elucidate how the antigenic impacts of SARS-CoV-2 mutations depend on exposure history.
]]></description>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Eguia, R. T.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Khan, K.</dc:creator>
<dc:creator>Franko, N.</dc:creator>
<dc:creator>Logue, J. K.</dc:creator>
<dc:creator>Lord, S. M.</dc:creator>
<dc:creator>Speake, C.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Sigal, A.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2022-03-14</dc:date>
<dc:identifier>doi:10.1101/2022.03.12.484088</dc:identifier>
<dc:title><![CDATA[The SARS-CoV-2 Delta variant induces an antibody response largely focused on class 1 and 2 antibody epitopes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.12.484096v1?rss=1">
<title>
<![CDATA[
The cancer chemotherapeutic 5-fluorouracil is a potent inhibitor of Fusobacterium nucleatum and its activity is modified by the intratumoral microbiota 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.12.484096v1?rss=1</link>
<description><![CDATA[
Fusobacterium nucleatum is among the most prevalent and dominant bacterial species in colorectal cancer (CRC) tumor tissue, and growing evidence supports its role in cancer progression and poorer patient prognosis. Here we perform a small molecule inhibitor screen of 1,846 bioactive compounds against a CRC isolate of F. nucleatum and find that 15% of inhibitors are antineoplastic agents including fluoropyrimidines. Validation of these findings reveal that 5-fluorouracil (5-FU), the first-line chemotherapeutic used to treat CRC worldwide, is a potent inhibitor of F. nucleatum CRC isolates at concentrations found in serum of CRC patients treated with 5-FU. We also identify members of the intratumoral microbiota that are resistant to 5-FU, including Escherichia coli. Further, CRC E. coli isolates can modify 5-FU and relieve 5-FU toxicity towards otherwise-sensitive F. nucleatum and human CRC epithelial cells. Lastly, we demonstrate that ex-vivo CRC tumor microbiota from patients undergo different levels of community disruption after 5-FU exposure and have the potential to deplete 5-FU levels, thereby reducing local drug efficacy. Together, these observations argue for further investigation into the role that the CRC intratumoral microbiota plays in patient response to chemotherapy.
]]></description>
<dc:creator>LaCourse, K. D.</dc:creator>
<dc:creator>Baryiames, A.</dc:creator>
<dc:creator>Kempchinsky, A. G.</dc:creator>
<dc:creator>Johnston, C. D.</dc:creator>
<dc:creator>Bullman, S.</dc:creator>
<dc:date>2022-03-13</dc:date>
<dc:identifier>doi:10.1101/2022.03.12.484096</dc:identifier>
<dc:title><![CDATA[The cancer chemotherapeutic 5-fluorouracil is a potent inhibitor of Fusobacterium nucleatum and its activity is modified by the intratumoral microbiota]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.14.484352v1?rss=1">
<title>
<![CDATA[
Mitochondrial Redox Adaptations Enable Aspartate Synthesis in SDH-deficient Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.14.484352v1?rss=1</link>
<description><![CDATA[
The oxidative tricarboxylic acid (TCA) cycle is a central mitochondrial pathway integrating catabolic conversions of NAD+ to NADH and anabolic production of aspartate, a key amino acid for cell proliferation. Several TCA cycle components are implicated in tumorigenesis, including loss of function mutations in subunits of succinate dehydrogenase (SDH), also known as complex II of the electron transport chain (ETC). Mechanistic understanding of how proliferating cells tolerate the metabolic defects of SDH loss is still lacking. Here, we identify that SDH supports cell proliferation through aspartate synthesis but, unlike other ETC impairments, is not restored by electron acceptor supplementation. Interestingly, we find aspartate production and cell proliferation are restored to SDH impaired cells by concomitant inhibition of ETC complex I (CI). We determine that the benefits of CI inhibition in this context are dependent on decreasing mitochondrial NAD+/NADH, which drives SDH-independent aspartate production. We also find that genetic loss or restoration of SDH selects for cells with concordant CI activity, establishing distinct modalities of mitochondrial metabolism for maintaining aspartate synthesis. Collectively, these data identify a metabolically beneficial mechanism for CI loss in proliferating cells and reveal that compartmentalized redox changes can impact cellular fitness.
]]></description>
<dc:creator>Hart, M. L.</dc:creator>
<dc:creator>Quon, E.</dc:creator>
<dc:creator>Vigil, A.-L. B. G.</dc:creator>
<dc:creator>Engstrom, I. A.</dc:creator>
<dc:creator>Newsom, O. J.</dc:creator>
<dc:creator>Davidsen, K.</dc:creator>
<dc:creator>Hoellerbauer, P.</dc:creator>
<dc:creator>Carlisle, S. M.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:date>2022-03-14</dc:date>
<dc:identifier>doi:10.1101/2022.03.14.484352</dc:identifier>
<dc:title><![CDATA[Mitochondrial Redox Adaptations Enable Aspartate Synthesis in SDH-deficient Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.25.485875v1?rss=1">
<title>
<![CDATA[
Mosaic RBD nanoparticles protect against multiple sarbecovirus challenges in animal models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.25.485875v1?rss=1</link>
<description><![CDATA[
To combat future SARS-CoV-2 variants and spillovers of SARS-like betacoronaviruses (sarbecoviruses) threatening global health, we designed mosaic nanoparticles presenting randomly-arranged sarbecovirus spike receptor-binding domains (RBDs) to elicit antibodies against conserved/relatively-occluded, rather than variable/immunodominant/exposed, epitopes. We compared immune responses elicited by mosaic-8 (SARS-CoV-2 and seven animal sarbecoviruses) and homotypic (only SARS-CoV-2) RBD-nanoparticles in mice and macaques, observing stronger responses elicited by mosaic-8 to mismatched (not on nanoparticles) strains including SARS-CoV and animal sarbecoviruses. Mosaic-8 immunization showed equivalent neutralization of SARS-CoV-2 variants including Omicron and protected from SARS-CoV-2 and SARS-CoV challenges, whereas homotypic SARS-CoV-2 immunization protected only from SARS-CoV-2 challenge. Epitope mapping demonstrated increased targeting of conserved epitopes after mosaic-8 immunization. Together, these results suggest mosaic-8 RBD-nanoparticles could protect against SARS-CoV-2 variants and future sarbecovirus spillovers.
]]></description>
<dc:creator>Cohen, A. A.</dc:creator>
<dc:creator>van Doremalen, N. A.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Andersen, H.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Fan, C.</dc:creator>
<dc:creator>Schulz, J. E.</dc:creator>
<dc:creator>Gnanapragasam, P. N. P.</dc:creator>
<dc:creator>Kakutani, L. M.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Saturday, G.</dc:creator>
<dc:creator>Lee, Y. E.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Jette, C. A.</dc:creator>
<dc:creator>Lewis, M. G.</dc:creator>
<dc:creator>Tan, T. K.</dc:creator>
<dc:creator>Townsend, A. R.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Munster, V. J.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2022-03-28</dc:date>
<dc:identifier>doi:10.1101/2022.03.25.485875</dc:identifier>
<dc:title><![CDATA[Mosaic RBD nanoparticles protect against multiple sarbecovirus challenges in animal models]]></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/10.1101/2022.03.31.486605v1?rss=1">
<title>
<![CDATA[
Cell type-specific Expression Quantitative Trait Loci 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.31.486605v1?rss=1</link>
<description><![CDATA[
Mapping cell type-specific gene expression quantitative trait loci (ct-eQTLs) is a powerful way to investigate the genetic basis of complex traits. A popular method for ct-eQTL mapping is to assess the interaction between the genotype of a genetic locus and the abundance of a specific cell type using a linear model. However, this approach requires transforming RNA-seq count data, which distorts the relation between gene expression and cell type proportions and results in reduced power and/or inflated type I error. To address this issue, we have developed a statistical method called CSeQTL that allows for ct-eQTL mapping using bulk RNA-seq count data while taking advantage of allele-specific expression. We validated the results of CSeQTL through simulations and real data analysis, comparing CSeQTL results to those obtained from purified bulk RNA-seq data or single cell RNA-seq data. Using our ct-eQTL findings, we were able to identify cell types relevant to 21 categories of human traits.
]]></description>
<dc:creator>Little, P.</dc:creator>
<dc:creator>Zhabotynsky, V.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Lin, D.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2022-04-01</dc:date>
<dc:identifier>doi:10.1101/2022.03.31.486605</dc:identifier>
<dc:title><![CDATA[Cell type-specific Expression Quantitative Trait Loci]]></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/10.1101/2022.04.02.486748v1?rss=1">
<title>
<![CDATA[
SEACells: Inference of transcriptional and epigenomic cellular states from single-cell genomics data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.02.486748v1?rss=1</link>
<description><![CDATA[
Metacells are cell groupings derived from single-cell sequencing data that represent highly granular, distinct cell states. Here, we present single-cell aggregation of cell-states (SEACells), an algorithm for identifying metacells; overcoming the sparsity of single-cell data, while retaining heterogeneity obscured by traditional cell clustering. SEACells outperforms existing algorithms in identifying accurate, compact, and well-separated metacells in both RNA and ATAC modalities across datasets with discrete cell types and continuous trajectories. We demonstrate the use of SEACells to improve gene-peak associations, compute ATAC gene scores and measure gene accessibility in each metacell. Metacell-level analysis scales to large datasets and are particularly well suited for patient cohorts, including facilitation of data integration. We use our metacells to reveal expression dynamics and gradual reconfiguration of the chromatin landscape during hematopoietic differentiation, and to uniquely identify CD4 T cell differentiation and activation states associated with disease onset and severity in a COVID-19 patient cohort.
]]></description>
<dc:creator>Persad, S.</dc:creator>
<dc:creator>Choo, Z.-N.</dc:creator>
<dc:creator>Dien, C.</dc:creator>
<dc:creator>Masilionis, I.</dc:creator>
<dc:creator>Chaligne, R.</dc:creator>
<dc:creator>Nawy, T.</dc:creator>
<dc:creator>Brown, C. C.</dc:creator>
<dc:creator>Pe'er, I.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:creator>Pe'er, D.</dc:creator>
<dc:date>2022-04-03</dc:date>
<dc:identifier>doi:10.1101/2022.04.02.486748</dc:identifier>
<dc:title><![CDATA[SEACells: Inference of transcriptional and epigenomic cellular states from single-cell genomics data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.02.486791v1?rss=1">
<title>
<![CDATA[
Integrative analysis of metabolite GWAS illuminates the molecular basis of pleiotropy and genetic correlation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.02.486791v1?rss=1</link>
<description><![CDATA[
Pleiotropy and genetic correlation are widespread features in GWAS, but they are often difficult to interpret at the molecular level. Here, we perform GWAS of 16 metabolites clustered at the intersection of amino acid catabolism, glycolysis, and ketone body metabolism in a subset of UK Biobank. We utilize the well-documented biochemistry jointly impacting these metabolites to analyze pleiotropic effects in the context of their pathways. Among the 213 lead GWAS hits, we find a strong enrichment for genes encoding pathway-relevant enzymes and transporters. We demonstrate that the effect directions of variants acting on biology between metabolite pairs often contrast with those of upstream or downstream variants as well as the polygenic background. Thus, we find that these outlier variants often reflect biology local to the traits. Finally, we explore the implications for interpreting disease GWAS, underscoring the potential of unifying biochemistry with dense metabolomics data to understand the molecular basis of pleiotropy in complex traits and diseases.
]]></description>
<dc:creator>Smith, C. J.</dc:creator>
<dc:creator>Sinnott-Armstrong, N.</dc:creator>
<dc:creator>Cichonska, A.</dc:creator>
<dc:creator>Julkunen, H.</dc:creator>
<dc:creator>Fauman, E.</dc:creator>
<dc:creator>Wurtz, P.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:date>2022-04-04</dc:date>
<dc:identifier>doi:10.1101/2022.04.02.486791</dc:identifier>
<dc:title><![CDATA[Integrative analysis of metabolite GWAS illuminates the molecular basis of pleiotropy and genetic correlation]]></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/10.1101/2022.04.05.487100v1?rss=1">
<title>
<![CDATA[
The E2 glycoprotein holds key residues for Mayaro virus adaptation to the urban Aedes aegypti mosquito 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.05.487100v1?rss=1</link>
<description><![CDATA[
Adaptation to mosquito vectors suited for transmission in urban settings is a major driver in the emergence of arboviruses. To better anticipate future emergence events, it is crucial to assess their potential to adapt to new vector hosts. In this work, we used two different experimental evolution approaches to study the adaptation process of an emerging alphavirus, Mayaro virus (MAYV), to Aedes aegypti, an urban mosquito vector of many arboviruses. We identified E2-T179N as a key mutation increasing MAYV replication in insect cells and enhancing transmission by live Aedes aegypti. In contrast, this mutation decreased viral replication and binding in human fibroblasts, a primary cellular target of MAYV in humans. We also showed that MAYV E2-T179N was attenuated in vivo in a mouse model. We then used structural and experimental analyses to show that MAYV E2-T179N bound less efficiently to human cells, though the decrease in replication or binding was not mediated through interaction with one of the host receptors, Mxra8. When this mutation was introduced in the closely related chikungunya virus, which has caused major outbreaks globally in the past two decades, we observed increased replication in both human and insect cells, suggesting E2 position 179 is an important determinant of alphavirus host-adaptation, although in a virus-specific manner. Collectively, these results indicate that adaptation at the T179 residue in MAYV E2 may result in increased vector competence - but coming at the cost of optimal replication in humans - and may represent a first step towards a future emergence event.

Author SummaryMosquito-borne viruses must replicate in both mosquito and vertebrate hosts to be maintained in nature successfully. When viruses that are typically transmitted by forest dwelling mosquitoes enter urban environments due to deforestation or travel, they must adapt to urban mosquito vectors to transmit effectively. For mosquito-borne viruses, the need to also replicate in a vertebrate host like humans constrains this adaptation process. Towards understanding how the emerging alphavirus, Mayaro virus, might adapt to transmission by the urban mosquito vector, Aedes aegypti, we used natural evolution approaches to identify several viral mutations that impacted replication in both mosquito and vertebrate hosts. We show that a single mutation in the receptor binding protein increased transmission by Aedes aegypti but simultaneously reduced replication and disease in a mouse model, suggesting that this mutation alone is unlikely to be maintained in a natural transmission cycle between mosquitoes and humans. Understanding the adaptive potential of emerging viruses is critical to preventing future pandemics.
]]></description>
<dc:creator>Roesch, F.</dc:creator>
<dc:creator>Cereghino, C.</dc:creator>
<dc:creator>Carrau, L.</dc:creator>
<dc:creator>Hardy, A.</dc:creator>
<dc:creator>Ribeiro-Filho, H.</dc:creator>
<dc:creator>Henrion Lacritick, A.</dc:creator>
<dc:creator>Koh, C.</dc:creator>
<dc:creator>Marano, J.</dc:creator>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Rai, P.</dc:creator>
<dc:creator>Chuong, C.</dc:creator>
<dc:creator>Akter, S.</dc:creator>
<dc:creator>Vallet, T.</dc:creator>
<dc:creator>Blanc, H.</dc:creator>
<dc:creator>Elliot, T.</dc:creator>
<dc:creator>Brown, A. M.</dc:creator>
<dc:creator>Michalak, P.</dc:creator>
<dc:creator>LeRoith, T.</dc:creator>
<dc:creator>Bloom, J.</dc:creator>
<dc:creator>Marques, R. E.</dc:creator>
<dc:creator>Saleh, M.-C.</dc:creator>
<dc:creator>Vignuzzi, M.</dc:creator>
<dc:creator>Weger-Lucarelli, J.</dc:creator>
<dc:date>2022-04-05</dc:date>
<dc:identifier>doi:10.1101/2022.04.05.487100</dc:identifier>
<dc:title><![CDATA[The E2 glycoprotein holds key residues for Mayaro virus adaptation to the urban Aedes aegypti mosquito]]></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/10.1101/2022.04.05.487110v1?rss=1">
<title>
<![CDATA[
Distinct regions of H. pylori's bactofilin regulate protein interactions to control cell shape 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.05.487110v1?rss=1</link>
<description><![CDATA[
The helical shape of H. pylori cells promotes robust stomach colonization, however, how the helical shape of H. pylori cells is determined is unresolved. Previous work identified helical-cell-shape-promoting protein complexes containing a peptidoglycan-hydrolase (Csd1), a peptidoglycan precursor synthesis enzyme (MurF), a non-enzymatic homologue of Csd1 (Csd2), non-enzymatic transmembrane proteins (Csd5 and Csd7), and a bactofilin (CcmA). Bactofilins are highly conserved, spontaneously polymerizing cytoskeletal bacterial proteins. We sought to understand CcmAs function in generating the helical shape of H. pylori cells. Using CcmA deletion analysis, in vitro polymerization, and in vivo co-immunoprecipitation experiments we identified that the bactofilin domain and N-terminal region of CcmA are required for helical cell shape and the bactofilin domain of CcmA is sufficient for polymerization and interactions with Csd5 and Csd7. We also found that CcmAs N-terminal region inhibits interaction with Csd7. Deleting the N-terminal region of CcmA increases CcmA-Csd7 interactions and destabilizes the peptidoglycan-hydrolase Csd1. Using super-resolution microscopy, we found that Csd5 recruits CcmA to the cell envelope and promotes CcmA enrichment at the major helical axis. Thus, CcmA helps organize cell-shape-determining proteins and peptidoglycan synthesis machinery to coordinate cell wall modification and synthesis, promoting the curvature required to build a helical cell.
]]></description>
<dc:creator>Sichel, S. R.</dc:creator>
<dc:creator>Bratton, B. P.</dc:creator>
<dc:creator>Salama, N.</dc:creator>
<dc:date>2022-04-05</dc:date>
<dc:identifier>doi:10.1101/2022.04.05.487110</dc:identifier>
<dc:title><![CDATA[Distinct regions of H. pylori's bactofilin regulate protein interactions to control cell shape]]></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/10.1101/2022.04.11.487879v1?rss=1">
<title>
<![CDATA[
Broadly neutralizing antibodies target the coronavirus fusion peptide 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.11.487879v1?rss=1</link>
<description><![CDATA[
The potential for future coronavirus outbreaks highlights the need to develop strategies and tools to broadly target this group of pathogens. Here, using an epitope-agnostic approach, we identified six monoclonal antibodies that bound to spike proteins from all seven human-infecting coronaviruses. Epitope mapping revealed that all six antibodies target the conserved fusion peptide region adjacent to the S2 cleavage site. Two antibodies, COV44-62 and COV44-79, broadly neutralize a range of alpha and beta coronaviruses, including SARS-CoV-2 Omicron subvariants BA.1 and BA.2, albeit with lower potency than RBD-specific antibodies. In crystal structures of Fabs COV44-62 and COV44-79 with the SARS-CoV-2 fusion peptide, the fusion peptide epitope adopts a helical structure and includes the arginine at the S2 cleavage site. Importantly, COV44-79 limited disease caused by SARS-CoV-2 in a Syrian hamster model. These findings identify the fusion peptide as the target of the broadest neutralizing antibodies in an epitope-agnostic screen, highlighting this site as a candidate for next-generation coronavirus vaccine development.

One-Sentence SummaryRare monoclonal antibodies from COVID-19 convalescent individuals broadly neutralize coronaviruses by targeting the fusion peptide.
]]></description>
<dc:creator>Dacon, C.</dc:creator>
<dc:creator>Tucker, C.</dc:creator>
<dc:creator>Peng, L.</dc:creator>
<dc:creator>Lee, C.-C. D.</dc:creator>
<dc:creator>Lin, T.-H.</dc:creator>
<dc:creator>Yuan, M.</dc:creator>
<dc:creator>Cong, Y.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Purser, L.</dc:creator>
<dc:creator>Williams, J. K.</dc:creator>
<dc:creator>Pyo, C.-W.</dc:creator>
<dc:creator>Kosik, I.</dc:creator>
<dc:creator>Hu, Z.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Mohan, D.</dc:creator>
<dc:creator>Cooper, A.</dc:creator>
<dc:creator>Peterson, M.</dc:creator>
<dc:creator>Skinner, J.</dc:creator>
<dc:creator>Dixit, S.</dc:creator>
<dc:creator>Kollins, E.</dc:creator>
<dc:creator>Huzella, L.</dc:creator>
<dc:creator>Perry, D.</dc:creator>
<dc:creator>Byrum, R.</dc:creator>
<dc:creator>Lembirik, S.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Yang, E. S.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Leung, K.</dc:creator>
<dc:creator>Weinberg, R. S.</dc:creator>
<dc:creator>Pegu, A.</dc:creator>
<dc:creator>Geraghty, D.</dc:creator>
<dc:creator>Davidson, E.</dc:creator>
<dc:creator>Douagi, I.</dc:creator>
<dc:creator>Moir, S.</dc:creator>
<dc:creator>Yewdell, J. W.</dc:creator>
<dc:creator>Schmaljohn, C.</dc:creator>
<dc:creator>Crompton, P. D.</dc:creator>
<dc:creator>Holbrook, M. R.</dc:creator>
<dc:creator>Nemazee, D.</dc:creator>
<dc:creator>Mascola, J. R.</dc:creator>
<dc:creator>Wilson, I. A.</dc:creator>
<dc:creator>Tan, J.</dc:creator>
<dc:date>2022-04-12</dc:date>
<dc:identifier>doi:10.1101/2022.04.11.487879</dc:identifier>
<dc:title><![CDATA[Broadly neutralizing antibodies target the coronavirus fusion peptide]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.14.488385v1?rss=1">
<title>
<![CDATA[
Adaptive immunity is required for durable responses to alectinib in murine models of EML4-ALK lung cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.14.488385v1?rss=1</link>
<description><![CDATA[
PurposeLung cancers bearing oncogenic EML4-ALK fusions respond to targeted tyrosine kinase inhibitors (TKIs; e.g. alectinib), with variation in the degree of shrinkage and duration of treatment (DOT). We previously demonstrated a positive association of a TKI-induced interferon gamma (IFN{gamma}) transcriptional response with DOT in EGFR-mutant lung cancers. Herein, we used murine models of EML4-ALK lung cancer to test a role for host immunity in the therapeutic response to alectinib.

Experimental DesignThree murine EML4-ALK cell lines (EA1, EA2, EA3) were implanted orthotopically into the lungs of immunocompetent and immunodeficient mice and treated with alectinib. Tumor volumes were serially measured by CT. Immune cell content was measured by flow cytometry, multispectral immunofluorescence and CyTOF. Transcriptional responses to alectinib were assessed by RNAseq and secreted chemokines were measured by ELISA.

ResultsAll cell lines were sensitive to alectinib in vitro. EA1 and EA3 tumors retained residual disease that rapidly progressed upon termination of treatment while EA2 tumors were eliminated by TKI treatment. Alectinib induced inflammatory transcriptional programs and multiple chemokines in all cell lines while untreated tumors exhibited distinct baseline chemokine expression patterns and content of CD8+ T cells and myeloid subsets. When propagated in immune-deficient mice, all three cell line-derived lung tumor models exhibited significant shrinkage followed by prompt progression despite continuous alectinib treatment.

ConclusionsThe findings support an hypothesis that host and TKI-stimulated production of chemokines by tumor cells promotes functional engagement of adaptive immune cells within the tumor microenvironment that enhances the durability and depth of TKI response.

Statement of Translational RelevancePatients with metastatic lung cancer harboring ALK fusions are treated with targeted tyrosine kinase inhibitors (TKI) in the first line setting. Despite bearing the same driver oncogene, patients experience a range of tumor burden reduction and variable amounts of residual disease. Residual disease burden associates with patient survival and contributes to the emergence of drug resistance yielding treatment failure. The factors mediating this differential response to TKI and residual disease are incompletely understood. Our group has developed a panel of murine ALK driven lung cancer cell lines that reproducibly show differences in the depth and duration of response when implanted into immunocompetent mice. Data using this model indicate that the presence of CD8+ T cells is a major contributor to the depth and duration of response. These models will be critical in developing rational combination therapies to augment the immune microenvironment engagement along with TKIs to improve outcomes for these patients.
]]></description>
<dc:creator>Kleczko, E. K.</dc:creator>
<dc:creator>Hinz, T. K.</dc:creator>
<dc:creator>Nguyen, T. T.</dc:creator>
<dc:creator>Gurule, N. J.</dc:creator>
<dc:creator>Navarro, A.</dc:creator>
<dc:creator>Le, A. T.</dc:creator>
<dc:creator>Johnson, A. M.</dc:creator>
<dc:creator>Kwak, J.</dc:creator>
<dc:creator>Polhac, D. I.</dc:creator>
<dc:creator>Clambey, E. T.</dc:creator>
<dc:creator>Weiser-Evans, M. C.</dc:creator>
<dc:creator>Patil, T.</dc:creator>
<dc:creator>Schenk, E. L.</dc:creator>
<dc:creator>Heasley, L. E.</dc:creator>
<dc:creator>Nemenoff, R. A.</dc:creator>
<dc:date>2022-04-14</dc:date>
<dc:identifier>doi:10.1101/2022.04.14.488385</dc:identifier>
<dc:title><![CDATA[Adaptive immunity is required for durable responses to alectinib in murine models of EML4-ALK lung cancer]]></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/10.1101/2022.04.20.488892v1?rss=1">
<title>
<![CDATA[
The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.20.488892v1?rss=1</link>
<description><![CDATA[
Advances in genomic technology led to a more focused pattern for the distribution of chromosomal proteins and a better understanding of their functions. The recent development of the CUT&RUN technique marks one of the important such advances. Here we develop a modified CUT&RUN technique that we termed nanoCUT&RUN, in which a high affinity nanobody to GFP is used to bring micrococcal nuclease to the binding sites of GFP-tagged chromatin proteins. Subsequent activation of the nuclease cleaves the chromatin, and sequencing of released DNA identifies binding sites. We show that nanoCUT&RUN efficiently produces high quality data for the TRL transcription factor in Drosophila embryos, and distinguishes binding sites specific between two TRL isoforms. We further show that nanoCUT&RUN dissects the distributions of the HipHop and HOAP telomere capping proteins, and uncovers unexpected binding of telomeric proteins at centromeres. nanoCUT&RUN can be readily applied to any system in which a chromatin protein of interest, or its isoforms, carries the GFP tag.
]]></description>
<dc:creator>chen, t.</dc:creator>
<dc:creator>Wei, X.</dc:creator>
<dc:creator>Courret, C.</dc:creator>
<dc:creator>cui, m.</dc:creator>
<dc:creator>Cheng, L.</dc:creator>
<dc:creator>wu, j.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Larracuente, A. M.</dc:creator>
<dc:creator>rong, y.</dc:creator>
<dc:date>2022-04-22</dc:date>
<dc:identifier>doi:10.1101/2022.04.20.488892</dc:identifier>
<dc:title><![CDATA[The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.20.488939v1?rss=1">
<title>
<![CDATA[
Transcriptional atlas of the human immune response to 13 vaccines reveals a common predictor of vaccine-induced antibody responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.20.488939v1?rss=1</link>
<description><![CDATA[
Systems biology approaches have been used to define molecular signatures and mechanisms of immunity to vaccination. However, most such studies have been done with single vaccines, and comparative analysis of the response to different vaccines is lacking. We integrated temporal transcriptional data of over 3,000 samples, obtained from 820 healthy adults across 28 studies of 13 different vaccines and analyzed vaccination-induced signatures associated with the antibody response. Most vaccines induced similar kinetics of shared transcriptional signatures, including signatures of innate immunity occurring 1-3 days post-vaccination, as well as the canonical plasmablast and cell cycle signatures appearing 7 days post-vaccination. However, the yellow fever vaccine YF-17D uniquely induced an early transient signature of T and B cell activation at Day 1, followed by delayed antiviral/interferon and plasmablast signatures that peaked at Days 7 and 14-21, respectively. Thus, despite the shared transcriptional response to most vaccines, at any given time point there was no evidence for a "universal signature" that could be used to predict the antibody response to all vaccines. However, accounting for the asynchronous nature of responses led to the identification of a time-adjusted signature that improved prediction antibody of responses across vaccines. These results provide a transcriptional atlas of the human immune response to vaccination and define a common, time-adjusted signature of antibody responses to vaccination.
]]></description>
<dc:creator>Hagan, T.</dc:creator>
<dc:creator>Gerristen, B.</dc:creator>
<dc:creator>Tomalin, L. E.</dc:creator>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Mule, M. P.</dc:creator>
<dc:creator>Chawla, D. G.</dc:creator>
<dc:creator>Rychkov, D.</dc:creator>
<dc:creator>Henrich, E.</dc:creator>
<dc:creator>Miller, H. E.</dc:creator>
<dc:creator>Diray-Arce, J.</dc:creator>
<dc:creator>Dunn, P.</dc:creator>
<dc:creator>Lee, A.</dc:creator>
<dc:creator>The Human Immunology Project Consortium (HIPC),</dc:creator>
<dc:creator>Levy, O.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Sarwal, M. M.</dc:creator>
<dc:creator>Tsang, J.</dc:creator>
<dc:creator>Suarez-Farinas, M.</dc:creator>
<dc:creator>Sekaly, R.-P.</dc:creator>
<dc:creator>Kleinstein, S.</dc:creator>
<dc:creator>Pulendran, B.</dc:creator>
<dc:date>2022-04-26</dc:date>
<dc:identifier>doi:10.1101/2022.04.20.488939</dc:identifier>
<dc:title><![CDATA[Transcriptional atlas of the human immune response to 13 vaccines reveals a common predictor of vaccine-induced antibody responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.23.488844v1?rss=1">
<title>
<![CDATA[
Evolutionarily conserved effects of Notch signaling drive intestinal graft-versus-host disease in mice and non-human primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.23.488844v1?rss=1</link>
<description><![CDATA[
Notch signaling promotes T-cell pathogenicity and graft-versus-host disease (GVHD) after allogeneic hematopoietic cell transplantation (allo-HCT) in mice, with a dominant role for the Delta-like ligand DLL4. To assess if Notchs effects are evolutionarily conserved and identify key mechanisms, we studied antibody-mediated DLL4 blockade in a non-human primate model similar to human allo-HCT. Short-term DLL4 blockade improved post-transplant survival with striking, durable protection from gastrointestinal GVHD, out of proportion to other disease sites. Unlike prior immunosuppressive strategies, anti-DLL4 interfered with a T-cell transcriptional program associated with intestinal infiltration. In cross-species investigations, Notch inhibition decreased surface abundance of the gut-homing integrin a4b7 in conventional T-cells via b1 competition for a4 binding, while preserving a4b7 in regulatory T-cells. Thereby, DLL4/Notch blockade decreased effector T-cell infiltration into the gut, with increased regulatory to conventional T-cell ratios early after allo-HCT. Our results identify a conserved, biologically unique and targetable role of DLL4/Notch signaling in GVHD.

One Sentence SummaryNotch signaling promotes pathogenic effector T cell infiltration of the intestine during acute graft-versus-host disease.
]]></description>
<dc:creator>Tkachev, V.</dc:creator>
<dc:creator>Vanderbeck, A.</dc:creator>
<dc:creator>Perkey, E.</dc:creator>
<dc:creator>Furlan, S. N.</dc:creator>
<dc:creator>McGuckin, C.</dc:creator>
<dc:creator>Gomez Atria, D.</dc:creator>
<dc:creator>Gerdemann, U.</dc:creator>
<dc:creator>Rui, X.</dc:creator>
<dc:creator>Lane, J.</dc:creator>
<dc:creator>Hunt, D. J.</dc:creator>
<dc:creator>Zheng, H.</dc:creator>
<dc:creator>Colonna, L.</dc:creator>
<dc:creator>Hoffman, M.</dc:creator>
<dc:creator>Yu, A.</dc:creator>
<dc:creator>Kelly, S.</dc:creator>
<dc:creator>Allman, A.</dc:creator>
<dc:creator>Burbach, B.</dc:creator>
<dc:creator>Shimizu, Y.</dc:creator>
<dc:creator>Panoskaltsis-Mortari, A.</dc:creator>
<dc:creator>Chen, G.</dc:creator>
<dc:creator>Carpenter, S.</dc:creator>
<dc:creator>Harari, O.</dc:creator>
<dc:creator>Kuhnert, F.</dc:creator>
<dc:creator>Thurston, G.</dc:creator>
<dc:creator>Blazar, B. R.</dc:creator>
<dc:creator>Kean, L. S.</dc:creator>
<dc:creator>Maillard, I.</dc:creator>
<dc:date>2022-04-27</dc:date>
<dc:identifier>doi:10.1101/2022.04.23.488844</dc:identifier>
<dc:title><![CDATA[Evolutionarily conserved effects of Notch signaling drive intestinal graft-versus-host disease in mice and non-human primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.04.490636v1?rss=1">
<title>
<![CDATA[
Evaluation of Cognitive Function in the Dog Aging Project: Associations with Baseline Canine Characteristics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.04.490636v1?rss=1</link>
<description><![CDATA[
Canine Cognitive Dysfunction (CCD) is a neurodegenerative disease in aging dogs. It has been described previously in relatively small cohorts of dogs using multiple different rating scales. This study aimed to use a minimally modified CCD rating scale developed by previous researchers to describe the prevalence of CCD more thoroughly in a large, nationwide cohort of companion dogs participating in the Dog Aging Project (DAP). Associations between various canine characteristics, predicted lifespan quartiles, and CCD were examined using univariable and multivariable logistic regression models and Receiver Operating Curve (ROC) analysis.

When controlling for all other characteristics, the odds of CCD increased 52% with each additional year of age. Among dogs of the same age, health status, breed type, and sterilization status, odds of CCD were 6.47 times higher in dogs who were not active compared to those who were very active. When controlling for age, breed type, activity level, and other comorbidities, dogs with a history of neurological, eye, or ear disorders had higher odds of CCD. Lifespan quartile analysis showed excellent discriminating ability between CCD positive and negative dogs. Weight-based lifespan quartile estimation could therefore serve as a tool to inform CCD screening by veterinarians.
]]></description>
<dc:creator>Yarborough, S.</dc:creator>
<dc:creator>Fitzpatrick, A.</dc:creator>
<dc:creator>Schwartz, S.</dc:creator>
<dc:creator>Dog Aging Project Consortium,</dc:creator>
<dc:date>2022-05-05</dc:date>
<dc:identifier>doi:10.1101/2022.05.04.490636</dc:identifier>
<dc:title><![CDATA[Evaluation of Cognitive Function in the Dog Aging Project: Associations with Baseline Canine Characteristics]]></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/10.1101/2022.05.09.491196v1?rss=1">
<title>
<![CDATA[
Persistent serum protein signatures define an inflammatory subset of long COVID 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.09.491196v1?rss=1</link>
<description><![CDATA[
Long COVID or post-acute sequelae of SARS-CoV-2 (PASC) is a clinical syndrome featuring diverse symptoms that can persist for months after acute SARS-CoV-2 infection. The etiologies are unknown but may include persistent inflammation, unresolved tissue damage, or delayed clearance of viral protein or RNA. Attempts to classify subsets of PASC by symptoms alone have been unsuccessful. To molecularly define PASC, we evaluated the serum proteome in longitudinal samples from 55 PASC individuals with symptoms lasting [&ge;]60 days after onset of acute infection and compared this to symptomatically recovered SARS-CoV-2 infected and uninfected individuals. We identified subsets of PASC with distinct signatures of persistent inflammation. Type II interferon signaling and canonical NF-{kappa}B signaling (particularly associated with TNF), were the most differentially enriched pathways. These findings help to resolve the heterogeneity of PASC, identify patients with molecular evidence of persistent inflammation, and highlight dominant pathways that may have diagnostic or therapeutic relevance.

One Sentence SummarySerum proteome profiling identifies subsets of long COVID patients with evidence of persistent inflammation including key immune signaling pathways that may be amenable to therapeutic intervention.
]]></description>
<dc:creator>Talla, A.</dc:creator>
<dc:creator>Vasaikar, S. V.</dc:creator>
<dc:creator>Szeto, G.</dc:creator>
<dc:creator>Lemos, M. P.</dc:creator>
<dc:creator>Czartoski, J. L.</dc:creator>
<dc:creator>MacMillan, H.</dc:creator>
<dc:creator>Moodie, Z.</dc:creator>
<dc:creator>Cohen, K. W.</dc:creator>
<dc:creator>Fleming, L. B.</dc:creator>
<dc:creator>Thomson, Z.</dc:creator>
<dc:creator>Okada, L.</dc:creator>
<dc:creator>Becker, L. A.</dc:creator>
<dc:creator>Coffey, E. M.</dc:creator>
<dc:creator>DeRosa, S. C.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Skene, P. J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Bumol, T. F.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Torgerson, T. R.</dc:creator>
<dc:date>2022-05-10</dc:date>
<dc:identifier>doi:10.1101/2022.05.09.491196</dc:identifier>
<dc:title><![CDATA[Persistent serum protein signatures define an inflammatory subset of long COVID]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.24.493335v1?rss=1">
<title>
<![CDATA[
Coenzyme A binding sites induce proximal acylation across protein families 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.24.493335v1?rss=1</link>
<description><![CDATA[
Lysine N{varepsilon}-acylations, such as acetylation or succinylation, are post-translational modifications that regulate protein function. In mitochondria, lysine acylation is predominantly non-enzymatic, and only a specific subset of the proteome is acylated. Coenzyme A (CoA) can act as an acyl group carrier via a thioester bond, but what controls the acylation of mitochondrial lysines remains poorly understood. Using published datasets, here we found that proteins with a CoA-binding site are more likely to be acetylated, succinylated, and glutarylated. Using computational modeling, we show that lysine residues near the CoA-binding pocket are highly acylated compared to those farther away. We hypothesized that acyl-CoA binding enhances acylation of nearby lysine residues. To test this hypothesis, we co-incubated enoyl-CoA hydratase short chain 1 (ECHS1), a CoA-binding mitochondrial protein, with succinyl-CoA and CoA. Using mass spectrometry, we found that succinyl-CoA induced widespread lysine succinylation and that CoA competitively inhibited ECHS1 succinylation. CoA-induced inhibition at a particular lysine site correlated inversely with the distance between that lysine and the CoA-binding pocket. Our study indicated that CoA acts as a competitive inhibitor of ECHS1 succinylation by binding to the CoA-binding pocket. Together, this suggests that proximal acylation at CoA-binding sites is a primary mechanism for lysine acylation in the mitochondria.
]]></description>
<dc:creator>Carrico, C.</dc:creator>
<dc:creator>Cruz, A.</dc:creator>
<dc:creator>Walter, M.</dc:creator>
<dc:creator>Meyer, J. G.</dc:creator>
<dc:creator>Wehrfritz, C.</dc:creator>
<dc:creator>Shah, S.</dc:creator>
<dc:creator>Wei, L.</dc:creator>
<dc:creator>Schilling, B.</dc:creator>
<dc:creator>Verdin, E.</dc:creator>
<dc:date>2022-05-25</dc:date>
<dc:identifier>doi:10.1101/2022.05.24.493335</dc:identifier>
<dc:title><![CDATA[Coenzyme A binding sites induce proximal acylation across protein families]]></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/10.1101/2022.05.31.493913v1?rss=1">
<title>
<![CDATA[
An in vivo biomarker to characterize ototoxic compounds and novel protective therapeutics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.31.493913v1?rss=1</link>
<description><![CDATA[
There are no approved therapeutics for the prevention of hearing loss and vestibular dysfunction from drugs like aminoglycoside antibiotics. While the mechanisms underlying aminoglycoside ototoxicity remain unresolved, there is considerable evidence that aminoglycosides enter inner ear mechanosensory hair cells through the mechanoelectrical transduction (MET) channel. Inhibition of MET-dependent uptake with small molecules or modified aminoglycosides is a promising otoprotective strategy. To better characterize mammalian ototoxicity and aid in the translation of emerging therapeutics, a biomarker is needed. In the present study we propose that neonatal mice systemically injected with the aminoglycosides G418 conjugated to Texas Red (G418-TR) can be used as a histologic biomarker to characterize in vivo aminoglycoside toxicity. We demonstrate that postnatal day 5 mice, like older mice with functional hearing, show uptake and retention of G418-TR in cochlear hair cells following systemic injection. When we compare G418-TR uptake in other tissues, we find that kidney proximal tubule cells show similar retention. Using ORC-13661, an investigational hearing protection drug, we demonstrate in vivo inhibition of aminoglycoside uptake in mammalian hair cells. This work establishes how systemically administered fluorescently labeled ototoxins in the neonatal mouse can reveal important details about ototoxic drugs and protective therapeutics.
]]></description>
<dc:creator>Bellairs, J. A.</dc:creator>
<dc:creator>Redila, V.</dc:creator>
<dc:creator>Wu, P.</dc:creator>
<dc:creator>Tong, L.</dc:creator>
<dc:creator>Webster, A.</dc:creator>
<dc:creator>Simon, J. A.</dc:creator>
<dc:creator>Rubel, E. W.</dc:creator>
<dc:creator>Raible, D. W.</dc:creator>
<dc:date>2022-06-01</dc:date>
<dc:identifier>doi:10.1101/2022.05.31.493913</dc:identifier>
<dc:title><![CDATA[An in vivo biomarker to characterize ototoxic compounds and novel protective therapeutics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.31.494200v1?rss=1">
<title>
<![CDATA[
Androgen receptor inhibition induces metabolic reprogramming and increased reliance on oxidative mitochondrial metabolism in prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.31.494200v1?rss=1</link>
<description><![CDATA[
Prostate cancer cells that survive clinical androgen receptor (AR) blockade mediate disease progression and lethality. Reprogrammed metabolic signaling is one mechanism by which tumor cells can survive treatment. However, how AR inhibition reprograms metabolism, and whether altered metabolism can be exploited to eradicate cells that survive AR blockade, remains unclear. Here, we comprehensively characterized the effect of AR blockade on prostate cancer metabolism using transcriptomics, metabolomics, and bioenergetics approaches. AR inhibition maintains oxidative mitochondrial metabolism and reduces glycolytic signaling, through hexokinase II downregulation and decreased MYC activity. Robust elongation of mitochondria via reduced DRP1 activity supports cell fitness after AR blockade. In addition, AR inhibition enhances sensitivity to complex I inhibitors in several models, suggesting that AR blockade increases reliance on oxidative mitochondrial metabolism. Our study provides an enhanced understanding of how AR inhibition alters metabolic signaling and highlights the potential of therapies that target metabolic vulnerabilities in AR-inhibited cells.
]]></description>
<dc:creator>Crowell, P.</dc:creator>
<dc:creator>Giafaglione, J.</dc:creator>
<dc:creator>Jones, A.</dc:creator>
<dc:creator>Nunley, N.</dc:creator>
<dc:creator>Hashimoto, T.</dc:creator>
<dc:creator>Delcourt, A.</dc:creator>
<dc:creator>Petcherski, A.</dc:creator>
<dc:creator>Bernard, M.</dc:creator>
<dc:creator>Huang, R. R.</dc:creator>
<dc:creator>Low, J.-Y.</dc:creator>
<dc:creator>Matulionis, N.</dc:creator>
<dc:creator>Guan, X.</dc:creator>
<dc:creator>Navone, N.</dc:creator>
<dc:creator>Alumkal, J.</dc:creator>
<dc:creator>Haffner, M.</dc:creator>
<dc:creator>Ye, H.</dc:creator>
<dc:creator>Zoubeidi, A.</dc:creator>
<dc:creator>Christofk, H.</dc:creator>
<dc:creator>Shirihai, O. S.</dc:creator>
<dc:creator>Divakaruni, A. S.</dc:creator>
<dc:creator>Goldstein, A. S.</dc:creator>
<dc:date>2022-06-01</dc:date>
<dc:identifier>doi:10.1101/2022.05.31.494200</dc:identifier>
<dc:title><![CDATA[Androgen receptor inhibition induces metabolic reprogramming and increased reliance on oxidative mitochondrial metabolism in prostate cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.31.494218v1?rss=1">
<title>
<![CDATA[
HSV-1 exploits host heterochromatin for egress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.31.494218v1?rss=1</link>
<description><![CDATA[
Herpes simplex virus (HSV-1) progeny form in the nucleus and must exit to successfully infect other cells. These newly formed viral capsids navigate the complex chromatin architecture of the nucleus to reach the inner nuclear membrane and egress. Here, we demonstrate by transmission electron microscopy (TEM) that HSV-1 capsids traverse dense heterochromatin in the nuclear periphery to reach the inner nuclear membrane. We found that this heterochromatin is dependent on the specific chromatin marks of trimethylation on histone H3 lysine 27 (H3K27me3) and the histone variant macroH2A1. Through chromatin profiling over the course of infection, we revealed massive host genomic regions bound by macroH2A1 and H3K27me3 that correlate with decreased host transcription in active compartments. This indicates the formation of new heterochromatin during infection. We found that loss of these markers resulted in significantly lower viral titers but did not impact viral genome or protein accumulation. Strikingly, we discovered by TEM that loss of macroH2A1 or H3K27me3 resulted in nuclear trapping of viral capsids. Thus, our work demonstrates that HSV-1 takes advantage of the dynamic nature of host heterochromatin formation during infection for efficient viral egress.
]]></description>
<dc:creator>Lewis, H. C.</dc:creator>
<dc:creator>Kelnhofer-Millevolte, L. E.</dc:creator>
<dc:creator>Brinkley, M. R.</dc:creator>
<dc:creator>Arbach, H. E.</dc:creator>
<dc:creator>Arnold, E. A.</dc:creator>
<dc:creator>Ramachandran, S.</dc:creator>
<dc:creator>Avgousti, D. C.</dc:creator>
<dc:date>2022-05-31</dc:date>
<dc:identifier>doi:10.1101/2022.05.31.494218</dc:identifier>
<dc:title><![CDATA[HSV-1 exploits host heterochromatin for egress]]></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/10.1101/2022.06.02.493651v1?rss=1">
<title>
<![CDATA[
Distinct antibody responses to endemic coronaviruses pre- and post-SARS-CoV-2 infection in Kenyan infants and mothers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.02.493651v1?rss=1</link>
<description><![CDATA[
Pre-existing antibodies that bind endemic human coronaviruses (eHCoVs) can cross-react with SARS-CoV-2, the betacoronavirus that causes COVID-19, but whether these responses influence SARS-CoV-2 infection is still under investigation and is particularly understudied in infants. In this study, we measured eHCoV and SARS-CoV-1 IgG antibody titers before and after SARS-CoV-2 seroconversion in a cohort of Kenyan women and their infants. Pre-existing eHCoV antibody binding titers were not consistently associated with SARS-CoV-2 seroconversion in infants or mothers, though we observed a very modest association between pre-existing HCoV-229E antibody levels and lack of SARS-CoV-2 seroconversion in infants. After seroconversion to SARS-CoV-2, antibody binding titers to endemic betacoronaviruses HCoV-OC43 and HCoV-HKU1, and the highly pathogenic betacoronavirus SARS-CoV-1, but not endemic alphacoronaviruses HCoV-229E and HCoV-NL63, increased in mothers. However, eHCoV antibody levels did not increase following SARS-CoV-2 seroconversion in infants, suggesting the increase seen in mothers was not simply due to cross-reactivity to naively generated SARS-CoV-2 antibodies. In contrast, the levels of antibodies that could bind SARS-CoV-1 increased after SARS-CoV-2 seroconversion in both mothers and infants, both of whom are unlikely to have had a prior SARS-CoV-1 infection, supporting prior findings that SARS-CoV-2 responses cross-react with SARS-CoV-1. In summary, we find evidence for increased eHCoV antibody levels following SARS-CoV-2 seroconversion in mothers but not infants, suggesting eHCoV responses can be boosted by SARS-CoV-2 infection when a prior memory response has been established, and that pre-existing cross-reactive antibodies are not strongly associated with SARS-CoV-2 infection risk in mothers or infants.
]]></description>
<dc:creator>Stoddard, C. I.</dc:creator>
<dc:creator>Sung, K.</dc:creator>
<dc:creator>Ojee, E.</dc:creator>
<dc:creator>Adhiambo, J.</dc:creator>
<dc:creator>Begnel, E. R.</dc:creator>
<dc:creator>Slyker, J.</dc:creator>
<dc:creator>Gantt, S.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Kinuthia, J.</dc:creator>
<dc:creator>Wamalwa, D.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Lehman, D. A.</dc:creator>
<dc:date>2022-06-03</dc:date>
<dc:identifier>doi:10.1101/2022.06.02.493651</dc:identifier>
<dc:title><![CDATA[Distinct antibody responses to endemic coronaviruses pre- and post-SARS-CoV-2 infection in Kenyan infants and mothers]]></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/10.1101/2022.06.07.495180v1?rss=1">
<title>
<![CDATA[
State-dependent protein-protein interactions mediating 4-1BB CAR Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.07.495180v1?rss=1</link>
<description><![CDATA[
Cells rely on activity-dependent protein-protein interactions to convey biological signals, but the state-dependent interactome is notoriously cell-specific and undercharacterized1. In the case of chimeric antigen receptor (CAR) T cells containing a 4-1BB costimulatory domain, receptor engagement is thought to trigger the formation of protein complexes similar to those triggered by T cell receptor (TCR)-mediated signaling, but the number and type of protein-interaction-mediating binding domains differ between CARs and TCRs. Here, we performed co-immunoprecipitation mass spectrometry of a 2nd generation CD19-directed 4-1BB:zeta CAR (referred to as bb{zeta}CAR) and identified 67 proteins that increased their co-association after target engagement. We compared activity-induced TCR and CAR signalosomes using quantitative multiplex co-immunoprecipitation and showed that bb{zeta}CAR engagement leads to activation of two modules of protein interactions, one similar to TCR signaling that is more weakly engaged in bb{zeta}CAR vs. TCR, and one composed of TRAF signaling complexes that is not engaged by the TCR. Batch-to-batch and inter-individual variations in IL2 production correlated with differences in the magnitude of protein network activation. Future CAR T cell manufacturing protocols could measure, and eventually control, biological variation by monitoring these signalosome activation markers.

One Sentence SummaryWe define a network of protein interactions engaged by chimeric antigen receptors following target binding, and show that the magnitude of network activation correlates with IL-2 secretion, a proxy measure for CAR T cell function.
]]></description>
<dc:creator>Ritmeester-Loy, S. A.</dc:creator>
<dc:creator>Lautz, J. D.</dc:creator>
<dc:creator>Zhang-Wong, Y.</dc:creator>
<dc:creator>Gustafson, J. A.</dc:creator>
<dc:creator>Wilson, A. L.</dc:creator>
<dc:creator>Lin, C.</dc:creator>
<dc:creator>Gafken, P. R.</dc:creator>
<dc:creator>Jensen, M. C.</dc:creator>
<dc:creator>Orentas, R.</dc:creator>
<dc:creator>Smith, S. E.</dc:creator>
<dc:date>2022-06-09</dc:date>
<dc:identifier>doi:10.1101/2022.06.07.495180</dc:identifier>
<dc:title><![CDATA[State-dependent protein-protein interactions mediating 4-1BB CAR Signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.08.495379v1?rss=1">
<title>
<![CDATA[
Genetic conflicts between sex chromosomes drive expansion and loss of sperm nuclear basic protein genes in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.08.495379v1?rss=1</link>
<description><![CDATA[
Many animal species employ short, positively charged proteins, called sperm nuclear basic proteins (SNBPs) or protamines, for tighter packaging of genomes in sperm. SNBP repertoires differ dramatically across animal lineages and signatures of rapid evolution have been reported in mammals. Both sperm competition and meiotic drive between sex chromosomes have been proposed as causes of SNBP innovation. We used a phylogenomic approach to investigate SNBP diversification and its underlying causes in Drosophila species. We found unambiguous signatures of positive selection in most SNBP genes except for genes essential for male fertility in D. melanogaster. Unexpectedly, evolutionarily young SNBP genes are more likely to encode essential functions for fertility than ancient, conserved SNBP genes like CG30056, which we found is dispensable for male fertility despite universal retention in Drosophila species. We found 19 independent amplification events involving eight SNBP genes that occurred preferentially on sex chromosomes in 78 Drosophila species. Conversely, we found that otherwise-conserved SNBP genes were lost in the montium group of Drosophila species, coincident with an X-Y chromosomal fusion. Furthermore, SNBP genes that became linked to sex chromosomes via chromosomal fusions are prone to degenerate or relocate back to autosomes. We hypothesize that SNBP genes ancestrally encoded by autosomes suppress meiotic drive, whereas sex-chromosomal SNBP expansions directly participate in meiotic drive. X-Y fusions in the montium group render autosomal SNBPs dispensable by making X-versus-Y meiotic drive obsolete or costly. We conclude that SNBP rapid evolution is driven by genetic conflicts between sex chromosomes during spermatogenesis in Drosophila species.
]]></description>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2022-06-09</dc:date>
<dc:identifier>doi:10.1101/2022.06.08.495379</dc:identifier>
<dc:title><![CDATA[Genetic conflicts between sex chromosomes drive expansion and loss of sperm nuclear basic protein genes in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.10.495681v1?rss=1">
<title>
<![CDATA[
Adjuvants influence the maturation of VRC01-like antibodies during immunization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.10.495681v1?rss=1</link>
<description><![CDATA[
Once naive B cells expressing germline VRC01-class B cell receptors become activated by germline-targeting immunogens, they enter germinal centers and undergo affinity maturation. Booster immunizations with heterologous Envs are required for the full maturation of VRC01-class antibodies. Here, we examined whether and how three adjuvants, Poly(I:C), GLA-LSQ, or Rehydragel, that activate different pathways of the innate immune system, influence the rate and type of somatic mutations accumulated by VRC01-class BCRs that become activated by the germline-targeting 426c.Mod.Core immunogen and the heterologous HxB2.WT.Core booster immunogen. We report that although the adjuvant used had no influence on the durability of plasma antibody responses after the prime, it influenced the plasma VRC01 antibody titers after the boost and the accumulation of somatic mutations on the elicited VRC01 antibodies.

ONE SENTENCE SUMMARYVRC01-class BCRs with different somatic mutations are being selected depending on the adjuvant used during immunization
]]></description>
<dc:creator>Kudsen, M. L.</dc:creator>
<dc:creator>Agrawal, P.</dc:creator>
<dc:creator>MacCamy, A.</dc:creator>
<dc:creator>Parks, R. K.</dc:creator>
<dc:creator>Gray, M. D.</dc:creator>
<dc:creator>Takushi, B.</dc:creator>
<dc:creator>Khechaduri, A.</dc:creator>
<dc:creator>Coler, R.</dc:creator>
<dc:creator>LaBranche, C.</dc:creator>
<dc:creator>Montefiori, D.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:date>2022-06-13</dc:date>
<dc:identifier>doi:10.1101/2022.06.10.495681</dc:identifier>
<dc:title><![CDATA[Adjuvants influence the maturation of VRC01-like antibodies during immunization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.15.496302v1?rss=1">
<title>
<![CDATA[
An anti-steatosis response regulated by oleic acid through lipid droplet-mediated ERAD enhancement 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.15.496302v1?rss=1</link>
<description><![CDATA[
Although excessive lipid accumulation is a hallmark of obesity-related pathologies, some lipids are beneficial. Oleic acid (OA), the most abundant monounsaturated fatty acid (FA), promotes health and longevity. Here we show that OA benefits C. elegans by activating the endoplasmic reticulum (ER)-resident transcription factor SKN-1A (Nrf1/NFE2L1) in a lipid homeostasis response. SKN-1A/Nrf1 is cleared from the ER by the ER-associated degradation (ERAD) machinery and stabilized when proteasome activity is low, and canonically maintains proteasome homeostasis. Unexpectedly, OA increases nuclear SKN-1A levels independently of proteasome activity, through lipid droplet (LD)-mediated enhancement of ERAD. In turn, SKN-1A reduces steatosis by reshaping the lipid metabolism transcriptome, and mediates longevity from OA provided through endogenous accumulation, reduced H3K4 trimethylation, or dietary supplementation. Our findings reveal a surprising mechanism of FA signal transduction, and a lipid homeostasis pathway that provides strategies for opposing steatosis and aging, and may mediate benefits of the OA-rich Mediterranean diet.
]]></description>
<dc:creator>Castillo-Quan, J. I.</dc:creator>
<dc:creator>Steinbaugh, M. J.</dc:creator>
<dc:creator>Fernandez-Cardenas, L. P.</dc:creator>
<dc:creator>Pohl, N. K.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Zhu, F.</dc:creator>
<dc:creator>Moroz, N.</dc:creator>
<dc:creator>Teixeira, V.</dc:creator>
<dc:creator>Bland, M. S.</dc:creator>
<dc:creator>Lehrbach, N. J.</dc:creator>
<dc:creator>Mazzeo, L. E. M.</dc:creator>
<dc:creator>Teufl, M.</dc:creator>
<dc:creator>Blackwell, T. K.</dc:creator>
<dc:date>2022-06-16</dc:date>
<dc:identifier>doi:10.1101/2022.06.15.496302</dc:identifier>
<dc:title><![CDATA[An anti-steatosis response regulated by oleic acid through lipid droplet-mediated ERAD enhancement]]></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/10.1101/2022.06.17.496580v1?rss=1">
<title>
<![CDATA[
Co-immunization with pre-erythrocytic antigens alongside circumsporozoite protein can enhance sterile protection against Plasmodium sporozoite infection. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.17.496580v1?rss=1</link>
<description><![CDATA[
Malaria-causing Plasmodium parasites have a complex life cycle and present numerous antigen targets that may contribute to protective immune responses. The currently recommended vaccine--RTS,S--functions by targeting the P. falciparum circumsporozoite protein (CSP), which is the most abundant surface protein of the sporozoite form responsible for initiating infection of the human host. Despite showing only moderate efficacy, RTS,S has established a strong foundation for the development of next-generation subunit vaccines. Our previous work characterizing the sporozoite surface proteome identified additional non-CSP antigens that may be useful as immunogens individually or in combination with CSP. In this study, we examined eight such antigens using the rodent malaria parasite P. yoelii as a model system. We demonstrate that despite conferring weak protection individually, co-immunizing each of several of these antigens alongside CSP, could significantly enhance the sterile protection achieved by CSP immunization alone. Thus, our work provides compelling evidence that a multi-antigen pre-erythrocytic vaccine approach may enhance protection compared to CSP-only vaccines. This lays the groundwork for further studies aimed at testing the identified antigen combinations in human vaccination trials that assess efficacy with controlled human malaria infection.

ImportanceThe currently approved malaria vaccine targets a single parasite protein (CSP) and only results in partial protection. We tested several additional vaccine targets in combination with CSP to identify those that could enhance protection from infection upon challenge in the mouse malaria model. In identifying several such enhancing vaccine targets, our work indicates that a multi-protein immunization approach may be a promising avenue to achieving higher levels of protection from infection. Our work identified several candidate leads for follow-up in the models relevant for human malaria, and provides an experimental framework for efficiently carrying out such screens for other combinations of vaccine targets.
]]></description>
<dc:creator>Vigdorovich, V.</dc:creator>
<dc:creator>Patel, H.</dc:creator>
<dc:creator>Watson, A.</dc:creator>
<dc:creator>Raappana, A.</dc:creator>
<dc:creator>Reynolds, L.</dc:creator>
<dc:creator>Selman, W.</dc:creator>
<dc:creator>Beeman, S.</dc:creator>
<dc:creator>Edlefsen, P. T.</dc:creator>
<dc:creator>Kappe, S. H. I.</dc:creator>
<dc:creator>Sather, D. N.</dc:creator>
<dc:date>2022-06-17</dc:date>
<dc:identifier>doi:10.1101/2022.06.17.496580</dc:identifier>
<dc:title><![CDATA[Co-immunization with pre-erythrocytic antigens alongside circumsporozoite protein can enhance sterile protection against Plasmodium sporozoite infection.]]></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/10.1101/2022.06.21.495546v1?rss=1">
<title>
<![CDATA[
Novel engineered chimeric engulfment receptors trigger T-cell effector functions against SIV infected CD4+ T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.21.495546v1?rss=1</link>
<description><![CDATA[
Adoptive therapy with genetically engineered T cells offers potential for infectious disease treatment in immunocompromised persons. HIV/simian immunodeficiency virus (SIV) infected cells express phosphatidylserine (PS) early post-infection. We tested whether chimeric engulfment receptor (CER) T cells designed to recognize PS-expressing cells could eliminate SIV infected cells. Lentiviral CER constructs comprised of the extracellular domain of T-cell immunoglobulin and mucin domain containing 4 (TIM-4), the PS receptor, and engulfment signaling domains were transduced into primary rhesus macaque (RM) T cells. We measured PS binding and T-cell engulfment of RM CD4+ T cells infected with SIV expressing GFP. As chimeric antigen receptor (CAR) T cells induce PS and subsequent TIM-4 binding, we evaluated in vitro killing of CAR and CER T-cell combinations. We found that recombinant TIM-4 bound to SIV infected cells. In vitro, CER CD4+ T cells effectively killed SIV infected cells, which was dependent on TIM-4 binding to PS. Enhanced killing of SIV infected CD4+ T cells by CER and CAR T-cell combinations was observed. This installation of innate immune functions into T cells presents an opportunity to enhance elimination of SIV infected cells and offers potential to augment functional cure of SIV/HIV infection.
]]></description>
<dc:creator>Corey, D.</dc:creator>
<dc:creator>Haeseleer, F.</dc:creator>
<dc:creator>Corey, L.</dc:creator>
<dc:date>2022-06-25</dc:date>
<dc:identifier>doi:10.1101/2022.06.21.495546</dc:identifier>
<dc:title><![CDATA[Novel engineered chimeric engulfment receptors trigger T-cell effector functions against SIV infected CD4+ T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.23.497404v1?rss=1">
<title>
<![CDATA[
The Functional Landscape of SARS-CoV-2 3CL Protease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.23.497404v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) as the etiologic agent of COVID-19 (coronavirus disease 2019) has drastically altered life globally. Numerous efforts have been placed on the development of therapeutics to treat SARS-CoV-2 infection. One particular target is the 3CL protease (3CLpro), which holds promise as it is essential to the virus and highly conserved among coronaviruses, suggesting that it may be possible to find broad inhibitors that treat not just SARS-CoV-2 but other coronavirus infections as well. While the 3CL protease has been studied by many groups for SARS-CoV-2 and other coronaviruses, our understanding of its tolerance to mutations is limited, knowledge which is particularly important as 3CL protease inhibitors become utilized clinically. Here, we develop a yeast-based deep mutational scanning approach to systematically profile the activity of all possible single mutants of the SARS-CoV-2 3CLpro, and validate our results both in yeast and in authentic viruses. We reveal that the 3CLpro is highly malleable and is capable of tolerating mutations throughout the protein, including within the substrate binding pocket. Yet, we also identify specific residues that appear immutable for function of the protease, suggesting that these interactions may be novel targets for the design of future 3CLpro inhibitors. Finally, we utilize our screening results as a basis to identify E166V as a resistance-conferring mutation against the therapeutic 3CLpro inhibitor, nirmatrelvir, in clinical use. Collectively, the functional map presented herein may serve as a guide for further understanding of the biological properties of the 3CL protease and for drug development for current and future coronavirus pandemics.
]]></description>
<dc:creator>Iketani, S.</dc:creator>
<dc:creator>Hong, S. J.</dc:creator>
<dc:creator>Sheng, J.</dc:creator>
<dc:creator>Bahari, F.</dc:creator>
<dc:creator>Culbertson, B.</dc:creator>
<dc:creator>Atanaki, F. F.</dc:creator>
<dc:creator>Aditham, A. K.</dc:creator>
<dc:creator>Kratz, A. F.</dc:creator>
<dc:creator>Luck, M. I.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Goff, S. P.</dc:creator>
<dc:creator>Montazeri, H.</dc:creator>
<dc:creator>Sabo, Y.</dc:creator>
<dc:creator>Ho, D. D.</dc:creator>
<dc:creator>Chavez, A.</dc:creator>
<dc:date>2022-06-24</dc:date>
<dc:identifier>doi:10.1101/2022.06.23.497404</dc:identifier>
<dc:title><![CDATA[The Functional Landscape of SARS-CoV-2 3CL Protease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.07.499178v1?rss=1">
<title>
<![CDATA[
A 96-WELL VALVED MICROFLUIDIC DEVICE FOR TESTING OF LIVE INTACT TUMOR CUBOIDS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.07.499178v1?rss=1</link>
<description><![CDATA[
There is a pressing need for functional testing platforms that use human, live tumor tissue to better predict traditional and immunotherapy responses. Such platforms should also retain as much of the native tumor microenvironment (TME) as possible, as many cancer drug actions rely on TME-dependent mechanisms. Present high-throughput testing platforms that have some of these features, e.g. based on patient-derived tumor organoids, require a growth step that alters the TME. On the other hand, micro-dissected tumor tissue "spheroids" that retain an intact TME have shown promising responses to immunomodulators acting on native immune cells. Here we demonstrate a microfluidic 96-well platform designed for drug treatment of hundreds of similarly-sized, cuboidal micro-tissues ("cuboids") produced from a single tumor sample. Four cuboids per well are automatically arrayed into the platform using hydrodynamic trapping. The microfluidic device, entirely fabricated in thermoplastics, features microvalves that fluidically isolate each well after the cuboid loading step. Since the platform effectively makes the most of scarce tumor tissue, we believe it could ultimately be applied to human biopsies for drug discovery and personalized oncology, altogether bypassing animal testing.
]]></description>
<dc:creator>Lockhart, E.</dc:creator>
<dc:creator>Horowitz, L. F.</dc:creator>
<dc:creator>Lim, C.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Mehrabi, M.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:creator>Folch, A.</dc:creator>
<dc:date>2022-07-08</dc:date>
<dc:identifier>doi:10.1101/2022.07.07.499178</dc:identifier>
<dc:title><![CDATA[A 96-WELL VALVED MICROFLUIDIC DEVICE FOR TESTING OF LIVE INTACT TUMOR CUBOIDS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.12.499365v1?rss=1">
<title>
<![CDATA[
Peptide binding specificity prediction using fine-tuned protein structure prediction networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.12.499365v1?rss=1</link>
<description><![CDATA[
Peptide binding proteins play key roles in biology, and predicting their binding specificity is a long-standing challenge. While considerable protein structural information is available, the most successful current methods use sequence information alone, in part because it has been a challenge to model the subtle structural changes accompanying sequence substitutions. Protein structure prediction networks such as AlphaFold model sequence-structure relationships very accurately, and we reasoned that if it were possible to specifically train such networks on binding data, more generalizable models could be created. We show that placing a classifier on top of the AlphaFold network and fine-tuning the combined network parameters for both classification and structure prediction accuracy leads to a model with strong generalizable performance on a wide range of Class I and Class II peptide-MHC interactions that approaches the overall performance of the state-of-the-art NetMHCpan sequence-based method. The peptide-MHC optimized model shows excellent performance in distinguishing binding and non-binding peptides to SH3 and PDZ domains. This ability to generalize well beyond the training set far exceeds that of sequence only models, and should be particularly powerful for systems where less experimental data is available.

Significance statementPeptide binding proteins carry out a variety of biological functions in cells and predicting their binding specificity could significantly improve our understanding of molecular pathways. Deep neural networks have achieved high structure prediction accuracy, but are not trained to predict binding specificity. Here we describe an approach to extending such networks to jointly predict protein structure and binding specificity. We incorporate AlphaFold into this approach, and fine-tune its parameters on peptide-MHC Class I and II structural and binding data. The fine-tuned model approaches state-of-the-art classification accuracy on peptide-MHC specificity prediction and generalizes to other peptide-binding systems such as the PDZ and SH3 domains.
]]></description>
<dc:creator>Motmaen, A.</dc:creator>
<dc:creator>Dauparas, J.</dc:creator>
<dc:creator>Baek, M.</dc:creator>
<dc:creator>Abedi, M.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:date>2022-07-13</dc:date>
<dc:identifier>doi:10.1101/2022.07.12.499365</dc:identifier>
<dc:title><![CDATA[Peptide binding specificity prediction using fine-tuned protein structure prediction networks]]></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/10.1101/2022.07.26.501561v1?rss=1">
<title>
<![CDATA[
Robust Harmonization of Microbiome Studies by Phylogenetic Scaffolding with MaLiAmPi 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.26.501561v1?rss=1</link>
<description><![CDATA[
Microbiome science is difficult to translate back to patients due to an inability to harmonize 16S rRNA gene-based microbiome data, as differences in the technique will result in different amplicon sequence variants (ASV) from the same microbe. Here we demonstrate that placement of ASV onto a common phylogenetic tree of full-length 16S rRNA alleles can harmonize microbiome studies. Using in silico data approximating 100 healthy human stool microbiomes we demonstrated that phylogenetic placement of ASV can recapitulate the true relationships between communities as compared closed-OTU based approaches (Spearman R 0.8 vs 0.2). Using real data from thousands of human gut and vaginal microbiota, we demonstrate phylogenetic placement, but not closed OTUs, were able to group communities by origin (stool vs vaginal) without being confounded by technique and integrate new data into existing ordination/clustering models for precision medicine. This enables meta-analysis of microbiome studies and the microbiome as a biomarker.
]]></description>
<dc:creator>Minot, S. S.</dc:creator>
<dc:creator>Garb, B.</dc:creator>
<dc:creator>Roldan, A.</dc:creator>
<dc:creator>Tang, A.</dc:creator>
<dc:creator>Oskotsky, T.</dc:creator>
<dc:creator>Rosenthal, C.</dc:creator>
<dc:creator>Hoffman, N. G.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:creator>Golob, J. L.</dc:creator>
<dc:date>2022-07-27</dc:date>
<dc:identifier>doi:10.1101/2022.07.26.501561</dc:identifier>
<dc:title><![CDATA[Robust Harmonization of Microbiome Studies by Phylogenetic Scaffolding with MaLiAmPi]]></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/10.1101/2022.07.29.501983v1?rss=1">
<title>
<![CDATA[
Beta human papillomavirus 8E6 promotes alternative end-joining 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.29.501983v1?rss=1</link>
<description><![CDATA[
Double strand breaks (DSBs) are one of the most lethal DNA lesions in cells. Previous studies show that the E6 protein of beta-human papillomavirus (HPV8 E6) impairs two major DSB repair pathways homologous recombination (HR) and non-homologous end-joining (NHEJ). However, HPV8 E6 delays but does not eliminate DSB repair capability of cells. How DSBs are repaired in cells with HPV8 E6 remains to be studied. We hypothesis that HPV8 E6 promotes a backup DSB repair pathway, alternative end-joining (Alt-EJ). Using CAS9 based Alt-EJ reporters, we show that HPV8 E6 promotes Alt-EJ. Further, using small molecule inhibitors, CRISPR/CAS9 gene knockout, and HPV8 E6 mutant, we find that HPV8 E6 promotes Alt-EJ by binding p300, an acetyltransferase that facilitates DSB repair by HR and NHEJ. Finally, we analyzed whole genome sequencing data from genomes of human foreskin keratinocytes expressing HPV8 E6 and found they displayed an increased frequency of deletions bearing the microhomology signatures of Alt-EJ. This study fills the knowledge gap how DSB is repaired in cells with HPV8 E6 and the mutagenic consequences of HPV8 E6 mediated p300 destabilization. Broadly, this study supports the hypothesis that beta-HPV promotes cancer formation by increasing genomic instability.
]]></description>
<dc:creator>Hu, C.</dc:creator>
<dc:creator>Bugbee, T.</dc:creator>
<dc:creator>Palinski, R.</dc:creator>
<dc:creator>Akinyemi, I.</dc:creator>
<dc:creator>McIntosh, M. T.</dc:creator>
<dc:creator>MacCarthy, T.</dc:creator>
<dc:creator>Bhaduri-McIntosh, S.</dc:creator>
<dc:creator>Wallace, N.</dc:creator>
<dc:date>2022-07-30</dc:date>
<dc:identifier>doi:10.1101/2022.07.29.501983</dc:identifier>
<dc:title><![CDATA[Beta human papillomavirus 8E6 promotes alternative end-joining]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.05.503004v1?rss=1">
<title>
<![CDATA[
Structure-based prediction of T cell receptor:peptide-MHC interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.05.503004v1?rss=1</link>
<description><![CDATA[
The regulatory and effector functions of T cells are initiated by the binding of their cell-surface T cell receptor (TCR) to peptides presented by major histocompatibility complex (MHC) proteins on other cells. The specificity of TCR:peptide-MHC interactions thus underlies nearly all adaptive immune responses. Despite intense interest, generalizable predictive models of TCR:peptide-MHC specificity remain out of reach; two key barriers are the diversity of TCR recognition modes and the paucity of training data. Inspired by recent breakthroughs in protein structure prediction achieved by deep neural networks, we evaluated structural modeling as a potential avenue for prediction of TCR epitope specificity. We show that a specialized version of the neural network predictor AlphaFold can generate models of TCR:peptide-MHC interactions that can be used to discriminate correct from incorrect peptide epitopes with substantial accuracy. Although much work remains to be done for these predictions to have widespread practical utility, we are optimistic that deep learning-based structural modeling represents a path to generalizable prediction of TCR:peptide-MHC interaction specificity.
]]></description>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:date>2022-08-06</dc:date>
<dc:identifier>doi:10.1101/2022.08.05.503004</dc:identifier>
<dc:title><![CDATA[Structure-based prediction of T cell receptor:peptide-MHC interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.09.503314v1?rss=1">
<title>
<![CDATA[
Human DUX4 and mouse Dux interact with STAT1 and broadly inhibit interferon-stimulated gene induction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.09.503314v1?rss=1</link>
<description><![CDATA[
DUX4 activates the first wave of zygotic gene expression in the early embryo. Mis-expression of DUX4 in skeletal muscle causes facioscapulohumeral dystrophy (FSHD), whereas expression in cancers suppresses IFN{gamma}-induction of MHC Class I and contributes to immune evasion. We show that the DUX4 protein interacts with STAT1 and broadly suppresses expression of IFN{gamma} stimulated genes by decreasing bound STAT1 and Pol-II recruitment. Transcriptional suppression of ISGs requires conserved (L)LxxL(L) motifs in the carboxyterminal region of DUX4 and phosphorylation of STAT1 Y701 enhances interaction with DUX4. Consistent with these findings, expression of endogenous DUX4 in FSHD muscle cells and the CIC-DUX4 fusion containing the DUX4 CTD in a sarcoma cell line inhibit IFN{gamma}-induction of ISGs. Mouse Dux similarly interacted with STAT1 and suppressed IFN{gamma} induction of ISGs. These findings identify an evolved role of the DUXC family in modulating immune signaling pathways with implications for development, cancers, and FSHD.
]]></description>
<dc:creator>Spens, A. E.</dc:creator>
<dc:creator>Sutliff, N. A.</dc:creator>
<dc:creator>Bennett, S. R.</dc:creator>
<dc:creator>Campbell, A. E.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:date>2022-08-11</dc:date>
<dc:identifier>doi:10.1101/2022.08.09.503314</dc:identifier>
<dc:title><![CDATA[Human DUX4 and mouse Dux interact with STAT1 and broadly inhibit interferon-stimulated gene induction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.20.504629v1?rss=1">
<title>
<![CDATA[
The Drosophila AWP1 ortholog Doctor No regulates JAK/STAT signaling for left-right asymmetry in the gut by promoting receptor endocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.20.504629v1?rss=1</link>
<description><![CDATA[
Many internal Drosophila organs show stereotypical left-right (LR) asymmetry, for which the underlying mechanisms remain elusive. Here, we identified an evolutionarily conserved ubiquitin-binding protein, AWP1/Doctor no (Drn), as a novel factor required for the LR asymmetry of the embryonic anterior gut in Drosophila. We showed that drn is essential in the circular visceral muscle cells of the midgut for JAK/STAT signaling, which contributes to the first known cue for anterior gut lateralization via LR-asymmetric nuclear rearrangement. Embryos homozygous for drn and lacking its maternal contribution showed phenotypes similar to that of depleted JAK/STAT signaling, suggesting that Drn is a general component of JAK/STAT signaling. The absence of Drn resulted in the specific accumulation of Domeless (Dome), the receptor of JAK/STAT signaling, in intracellular compartments. Thus, Drn is required for the endocytic trafficking of Dome, which is subsequently degraded in lysosomes. Our results suggest that the endocytosis of Dome is a critical step in activating JAK/STAT signaling. The roles of AWP1/Drn in activating JAK/STAT signaling and in LR-asymmetric development may be conserved in various organisms.

Summary StatementDr. No, a Drosophila ortholog of AWP1, activates JAK/STAT signaling via Dome receptor endocytosis in a crucial step for left-right asymmetry in the developing gut.
]]></description>
<dc:creator>Lai, Y.-T.</dc:creator>
<dc:creator>Sasamura, T.</dc:creator>
<dc:creator>Kuroda, J.</dc:creator>
<dc:creator>Maeda, R.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Hatori, R.</dc:creator>
<dc:creator>Ishibashi, T.</dc:creator>
<dc:creator>Taniguchi, K.</dc:creator>
<dc:creator>Ooike, M.</dc:creator>
<dc:creator>Taguchi, T.</dc:creator>
<dc:creator>Nakazawa, N.</dc:creator>
<dc:creator>Hozumi, S.</dc:creator>
<dc:creator>Okumura, T.</dc:creator>
<dc:creator>Aigaki, T.</dc:creator>
<dc:creator>Inaki, M.</dc:creator>
<dc:creator>Matsuno, K.</dc:creator>
<dc:date>2022-08-20</dc:date>
<dc:identifier>doi:10.1101/2022.08.20.504629</dc:identifier>
<dc:title><![CDATA[The Drosophila AWP1 ortholog Doctor No regulates JAK/STAT signaling for left-right asymmetry in the gut by promoting receptor endocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.29.505743v1?rss=1">
<title>
<![CDATA[
Molecular fate-mapping of serum antibodies reveals the effects of antigenic imprinting on repeated immunization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.29.505743v1?rss=1</link>
<description><![CDATA[
The ability of serum antibody to protect against pathogens arises from the interplay of antigen-specific B cell clones of different affinities and fine specificities. These cellular dynamics are ultimately responsible for serum-level phenomena such as antibody imprinting or "Original Antigenic Sin" (OAS), a proposed propensity of the immune system to rely repeatedly on the first cohort of B cells that responded to a stimulus upon exposure to related antigens. Imprinting/OAS is thought to pose a barrier to vaccination against rapidly evolving viruses such as influenza and SARS-CoV-2. Precise measurement of the extent to which imprinting/OAS inhibits the recruitment of new B cell clones by boosting is challenging because cellular and temporal origins cannot readily be assigned to antibodies in circulation. Thus, the extent to which imprinting/OAS impacts the induction of new responses in various settings remains unclear. To address this, we developed a "molecular fate-mapping" approach in which serum antibodies derived from specific cohorts of B cells can be differentially detected. We show that, upon sequential homologous boosting, the serum antibody response strongly favors reuse of the first cohort of B cell clones over the recruitment of new, naIve-derived B cells. This "primary addiction" decreases as a function of antigenic distance, allowing secondary immunization with divergent influenza virus or SARS-CoV-2 glycoproteins to overcome imprinting/OAS by targeting novel epitopes absent from the priming variant. Our findings have implications for the understanding of imprinting/OAS, and for the design and testing of vaccines aimed at eliciting antibodies to evolving antigens.
]]></description>
<dc:creator>Schiepers, A.</dc:creator>
<dc:creator>van 't Wout, M. F.</dc:creator>
<dc:creator>Greaney, A. J.</dc:creator>
<dc:creator>Zang, T.</dc:creator>
<dc:creator>Muramatsu, H.</dc:creator>
<dc:creator>Lin, P. J.</dc:creator>
<dc:creator>Tam, Y. K.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Pardi, N.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2022-08-30</dc:date>
<dc:identifier>doi:10.1101/2022.08.29.505743</dc:identifier>
<dc:title><![CDATA[Molecular fate-mapping of serum antibodies reveals the effects of antigenic imprinting on repeated immunization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.06.506695v1?rss=1">
<title>
<![CDATA[
High-risk microbial signatures are associated with severe parasitemia in controlled Plasmodium infections of both humans and rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.06.506695v1?rss=1</link>
<description><![CDATA[
While functions of the gastrointestinal (GI) microbiome include maintenance of immune homeostasis and protection against infectious disease, its role in determining disease severity during Plasmodium infection has been limited to mouse models and observational human cohorts. Here, we performed controlled Plasmodium infection in both humans and rhesus macaques (RMs) to experimentally determine the impact of GI microbiome composition on disease progression. Through analysis of serially collected microbiome samples, we identified a high-risk microbial signature that strongly associated with increased risk of developing severe parasitemia in human participants. Importantly, we identified a parallel phenomenon in RMs. The combined weight of this evidence demonstrates that pre-infection GI microbiome composition is highly indicative of P. falciparum disease risk. Moreover, our observation that P. fragile-microbiome dynamics in RMs closely mirrors P. falciparum-microbiome interactions in humans strongly supports the use of this model in pre-clinical investigations of novel microbiome-targeting approaches to reduce malaria burden.
]]></description>
<dc:creator>Gustin, A. T.</dc:creator>
<dc:creator>Broedlow, C. A.</dc:creator>
<dc:creator>Coronado, E.</dc:creator>
<dc:creator>Wangari, S.</dc:creator>
<dc:creator>Iwayama, N.</dc:creator>
<dc:creator>Ahrens, C. Y.</dc:creator>
<dc:creator>Garrison, W. D.</dc:creator>
<dc:creator>Guerriero, K. A.</dc:creator>
<dc:creator>De Paris, K.</dc:creator>
<dc:creator>Gale, M.</dc:creator>
<dc:creator>Klatt, N. R.</dc:creator>
<dc:creator>Kublin, J. G.</dc:creator>
<dc:creator>Manuzak, J. A.</dc:creator>
<dc:date>2022-09-07</dc:date>
<dc:identifier>doi:10.1101/2022.09.06.506695</dc:identifier>
<dc:title><![CDATA[High-risk microbial signatures are associated with severe parasitemia in controlled Plasmodium infections of both humans and rhesus macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.08.507086v1?rss=1">
<title>
<![CDATA[
CD4-binding site immunogens elicit heterologous anti-HIV-1 neutralizing antibodies in transgenic and wildtype animals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.08.507086v1?rss=1</link>
<description><![CDATA[
Passive transfer of broadly neutralizing anti-HIV-1 antibodies (bNAbs) protects against infection, and therefore eliciting bNAbs by vaccination is a major goal of HIV-1 vaccine efforts. bNAbs that target the CD4-binding site (CD4bs) on HIV-1 Env are among the most broadly active, but to date, responses elicited against this epitope in vaccinated animals have lacked potency and breadth. We hypothesized that CD4bs bNAbs resembling the antibody IOMA might be easier to elicit than other CD4bs antibodies that exhibit higher somatic mutation rates, a difficult-to-achieve mechanism to accommodate Envs N276gp120 N-glycan, and rare 5-residue light chain complementarity determining region 3s (CDRL3s). As an initial test of this idea, we developed IOMA germline-targeting Env immunogens and evaluated a sequential immunization regimen in transgenic mice expressing germline-reverted IOMA. These mice developed CD4bs epitope-specific responses with heterologous neutralization, and cloned antibodies overcame neutralization roadblocks including accommodating the N276gp120 glycan, with some neutralizing selected HIV-1 strains more potently than IOMA. The immunization regimen also elicited CD4bs-specific responses in animals containing polyclonal antibody repertoires. Thus, germline-targeting of IOMA-class antibody precursors represents a potential vaccine strategy to induce CD4bs bNAbs.
]]></description>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Loewe, M.</dc:creator>
<dc:creator>van Schooten, J.</dc:creator>
<dc:creator>Ramos, V.</dc:creator>
<dc:creator>Oliviera, T. Y.</dc:creator>
<dc:creator>Nishimura, Y.</dc:creator>
<dc:creator>Koranda, N. S.</dc:creator>
<dc:creator>Wall, A.</dc:creator>
<dc:creator>Yao, K.-H.</dc:creator>
<dc:creator>Poston, D.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Wiatr, M.</dc:creator>
<dc:creator>Horning, M.</dc:creator>
<dc:creator>Keeffe, J. R.</dc:creator>
<dc:creator>Hoffmann, M. A.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Abernathy, M. E.</dc:creator>
<dc:creator>Dam, K.-M. A.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Gnanapragasam, P. N.</dc:creator>
<dc:creator>Kakutani, L. M.</dc:creator>
<dc:creator>Pavlovitch-Bedzyk, A. J.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Howarth, M.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Martin, M. A.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:date>2022-09-12</dc:date>
<dc:identifier>doi:10.1101/2022.09.08.507086</dc:identifier>
<dc:title><![CDATA[CD4-binding site immunogens elicit heterologous anti-HIV-1 neutralizing antibodies in transgenic and wildtype animals]]></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/10.1101/2022.09.10.507413v1?rss=1">
<title>
<![CDATA[
The glutathione S-transferase Gstt1 is a robust driver of survival and dissemination in metastases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.10.507413v1?rss=1</link>
<description><![CDATA[
Identifying adaptive mechanisms of metastatic cancer cells remains an elusive question in the treatment of metastatic disease, particularly in pancreatic cancer (PDA), where the majority of patients present with metastatic lesions at the time of diagnosis. A loss-of-function shRNA targeted screen in metastatic-derived cells identified Gstt1, a member of the glutathione S-transferase superfamily, as uniquely required for metastasis and dissemination however dispensable for primary tumor growth. Gstt1 is expressed in early disseminated tumor cells (DTCs), is retained within a subpopulation of slow-cycling cells within established metastases and its inhibition led to a regression of macrometastatic lesions. This distinct Gstt1high population is highly metastatic and retains slow-cycling phenotypes, EMT features, and DTC characteristics compared to the Gstt1low population. Mechanistic studies indicate that in this subset of cells, Gstt1 maintains metastases by binding to and modifying intracellular fibronectin, regulating Fibronectin secretion from cancer cells and deposition into the metastatic microenvironment. We identified Gstt1 as a novel mediator of metastasis, highlighting the importance of metastatic heterogeneity and its influence on the metastatic tumor microenvironment.
]]></description>
<dc:creator>Ferrer, C. M.</dc:creator>
<dc:creator>Boon, R.</dc:creator>
<dc:creator>Cho, H.</dc:creator>
<dc:creator>Bernasocchi, T.</dc:creator>
<dc:creator>Wong, L. P.</dc:creator>
<dc:creator>Cetinbas, M.</dc:creator>
<dc:creator>Haggerty, E. R.</dc:creator>
<dc:creator>Mitsiades, I.</dc:creator>
<dc:creator>Wojtkiewicz, G. R.</dc:creator>
<dc:creator>McLoughlin, D. E.</dc:creator>
<dc:creator>Rheinbay, E.</dc:creator>
<dc:creator>Sadreyev, R.</dc:creator>
<dc:creator>Juric, D.</dc:creator>
<dc:creator>Mostoslavsky, R.</dc:creator>
<dc:date>2022-09-13</dc:date>
<dc:identifier>doi:10.1101/2022.09.10.507413</dc:identifier>
<dc:title><![CDATA[The glutathione S-transferase Gstt1 is a robust driver of survival and dissemination in metastases]]></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/10.1101/2022.09.13.507781v1?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/10.1101/2022.09.21.508922v1?rss=1">
<title>
<![CDATA[
Evolution of antibody immunity following Omicron BA.1 breakthrough infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.21.508922v1?rss=1</link>
<description><![CDATA[
Understanding the evolution of antibody immunity following heterologous SAR-CoV-2 breakthrough infection will inform the development of next-generation vaccines. Here, we tracked SARS-CoV-2 receptor binding domain (RBD)-specific antibody responses up to six months following Omicron BA.1 breakthrough infection in mRNA-vaccinated individuals. Cross-reactive serum neutralizing antibody and memory B cell (MBC) responses declined by two- to four-fold through the study period. Breakthrough infection elicited minimal de novo Omicron-specific B cell responses but drove affinity maturation of pre-existing cross-reactive MBCs toward BA.1. Public clones dominated the neutralizing antibody response at both early and late time points, and their escape mutation profiles predicted newly emergent Omicron sublineages. The results demonstrate that heterologous SARS-CoV-2 variant exposure drives the evolution of B cell memory and suggest that convergent neutralizing antibody responses continue to shape viral evolution.
]]></description>
<dc:creator>Kaku, C. I.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Dugan, H. L.</dc:creator>
<dc:creator>Khalife, P.</dc:creator>
<dc:creator>Song, G.</dc:creator>
<dc:creator>Champney, E. R.</dc:creator>
<dc:creator>Mielcarz, D. W.</dc:creator>
<dc:creator>Geoghegan, J. C.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Andrabi, R.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Walker, L. M.</dc:creator>
<dc:date>2022-09-22</dc:date>
<dc:identifier>doi:10.1101/2022.09.21.508922</dc:identifier>
<dc:title><![CDATA[Evolution of antibody immunity following Omicron BA.1 breakthrough infection]]></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/10.1101/2022.09.23.509057v1?rss=1">
<title>
<![CDATA[
AAV-delivered gene editing for latent genital or orofacial herpes simplex virus infection reduces ganglionic viral load and minimizes subsequent viral shedding in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.23.509057v1?rss=1</link>
<description><![CDATA[
Herpes simplex virus (HSV) establishes latency in ganglionic neurons of the peripheral nervous system, from which it can reactivate, causing recurrent disease and possible transmission to a new host. Current anti-HSV therapy does not eliminate latent HSV, and thus is only suppressive rather than curative. We developed a potentially curative approach to latent HSV infection and pathogenesis, based on gene editing using HSV-specific meganucleases delivered by adeno-associated virus (AAV) vectors. Our results demonstrated that a dual meganuclease therapy, composed of two anti-HSV-1 meganucleases delivered by a triple AAV serotype combination (AAV9, AAV-Dj/8, AAV-Rh10), can eliminate up to 97% of latent HSV DNA from ganglia in both ocular and vaginal mouse models of latent HSV infection. Using a novel pharmacological approach to reactivate latent HSV-1 in mice with the bromodomain inhibitor JQ-1, we demonstrated that this reduction in ganglionic viral load leads to a significant reduction of viral shedding from treated vs. control mice, with many treated mice showing no detectable virus shedding. In general, therapy was well tolerated, although dose-ranging studies showed hepatotoxicity at high AAV doses, consistent with previous observations in animals and humans. Also in agreement with previous literature, we observed subtle histological evidence of neuronal injury in some experimental mice, although none of the mice demonstrated observable neurological signs or deficits. These results reinforce the curative potential of gene editing for latent orofacial and genital HSV disease, and provide a framework for additional safety studies before human trials can begin.
]]></description>
<dc:creator>Aubert, M.</dc:creator>
<dc:creator>Haick, A. K.</dc:creator>
<dc:creator>Strongin, D. E.</dc:creator>
<dc:creator>Klouser, L. M.</dc:creator>
<dc:creator>Loprieno, M. A.</dc:creator>
<dc:creator>Stensland, L.</dc:creator>
<dc:creator>Santo, T. K.</dc:creator>
<dc:creator>Huang, M.-l.</dc:creator>
<dc:creator>Hyrien, O.</dc:creator>
<dc:creator>Stone, D.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:date>2022-09-26</dc:date>
<dc:identifier>doi:10.1101/2022.09.23.509057</dc:identifier>
<dc:title><![CDATA[AAV-delivered gene editing for latent genital or orofacial herpes simplex virus infection reduces ganglionic viral load and minimizes subsequent viral shedding in mice]]></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/10.1101/2022.10.06.511219v1?rss=1">
<title>
<![CDATA[
Improving polygenic risk prediction in admixed populations by explicitly modeling ancestral-specific effects via GAUDI 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.06.511219v1?rss=1</link>
<description><![CDATA[
Polygenic risk scores (PRS) have shown successes in clinics, but most PRS methods have focused only on individuals with one primary continental ancestry, thus poorly accommodating recently-admixed individuals. Here, we develop GAUDI, a novel penalized-regression-based method specifically designed for admixed individuals by explicitly modeling ancestry-specific effects and jointly estimating ancestry-shared effects. We demonstrate marked advantages of GAUDI over other methods through comprehensive simulation and real data analyses.
]]></description>
<dc:creator>Sun, Q.</dc:creator>
<dc:creator>Rowland, B. T.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Mikhaylova, A. V.</dc:creator>
<dc:creator>Avery, C.</dc:creator>
<dc:creator>Peters, U.</dc:creator>
<dc:creator>Lundin, J.</dc:creator>
<dc:creator>Matise, T.</dc:creator>
<dc:creator>Buyske, S.</dc:creator>
<dc:creator>Tao, R.</dc:creator>
<dc:creator>Mathias, R. A.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Auer, P. L.</dc:creator>
<dc:creator>Cox, N. J.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Thornton, T.</dc:creator>
<dc:creator>Raffield, L. M.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:date>2022-10-08</dc:date>
<dc:identifier>doi:10.1101/2022.10.06.511219</dc:identifier>
<dc:title><![CDATA[Improving polygenic risk prediction in admixed populations by explicitly modeling ancestral-specific effects via GAUDI]]></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/10.1101/2022.10.07.511358v1?rss=1">
<title>
<![CDATA[
Structural basis of epitope selectivity and potent protection from malaria by PfCSP antibody L9 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.07.511358v1?rss=1</link>
<description><![CDATA[
A primary objective in malaria vaccine design is the generation of high-quality antibody responses against the circumsporozoite protein of the malaria parasite, Plasmodium falciparum (PfCSP). To enable rational antigen design, we solved a cryo-EM structure of the highly potent anti-PfCSP antibody L9 in complex with recombinant PfCSP. We found that L9 Fab binds multivalently to the CSP minor (NPNV) repeats, which is stabilized by a novel set of affinity-matured homotypic, antibody-antibody contacts. Molecular dynamics simulations revealed a critical role of the L9 light chain in integrity of the homotypic interface, which likely impacts CSP affinity and protective efficacy. These findings reveal the molecular mechanism of the unique NPNV selectivity of L9 and emphasize the importance of anti-homotypic affinity maturation in protective immunity against P. falciparum.

One sentence summaryThe L9 light chain is crucial for potency by conferring multivalent, high affinity binding to the NPNV minor repeats of PfCSP.
]]></description>
<dc:creator>Martin, G.</dc:creator>
<dc:creator>Fernandez Quintero, M. L.</dc:creator>
<dc:creator>Lee, W.-H.</dc:creator>
<dc:creator>Pholcharee, T.</dc:creator>
<dc:creator>Eshun-Wilson, L.</dc:creator>
<dc:creator>Liedl, K. R.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>Seder, R. A.</dc:creator>
<dc:creator>Wilson, I. A.</dc:creator>
<dc:creator>Ward, A. B.</dc:creator>
<dc:date>2022-10-07</dc:date>
<dc:identifier>doi:10.1101/2022.10.07.511358</dc:identifier>
<dc:title><![CDATA[Structural basis of epitope selectivity and potent protection from malaria by PfCSP antibody L9]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.13.511039v1?rss=1">
<title>
<![CDATA[
ACSM1 and ACSM3 regulate prostate cancer fatty acid metabolism to promote tumour growth and constrain ferroptosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.13.511039v1?rss=1</link>
<description><![CDATA[
Prostate tumours are highly reliant on lipids for energy, growth and survival. Activity of the androgen receptor (AR) is associated with reprogramming of lipid metabolic processes in prostate cancer, although the molecular underpinnings of this relationship remain to be fully elucidated. Here, we identified Acyl-CoA Synthetase Medium Chain Family Members 1 and 3 (ACSM1 and ACSM3) as AR-regulated mediators of prostate cancer metabolism and growth. ACSM1 and ACSM3 are upregulated in prostate tumours compared to non-malignant tissues and other cancer types. Both enzymes enhanced proliferation and protected PCa cells from death in vitro, while silencing ACSM3 led to reduced tumour growth in an orthotopic xenograft model. We show that ACSM1 and ACSM3 are major regulators of the PCa lipidome and enhance energy production via fatty acid oxidation. Metabolic dysregulation caused by loss of ACSM1/3 led to mitochondrial oxidative stress, lipid peroxidation and cell death by ferroptosis. Conversely, over-expression of ACSM1/3 enabled PCa cells to survive toxic doses of medium chain fatty acids and promoted resistance to ferroptosis-inducing drugs and AR antagonists. Collectively, these studies uncover a new link between AR and lipid metabolism and position ACSM1 and ACSM3 as key players in prostate cancer progression and therapy resistance.
]]></description>
<dc:creator>Shrestha, R.</dc:creator>
<dc:creator>Nassar, Z. D.</dc:creator>
<dc:creator>Hanson, A. R.</dc:creator>
<dc:creator>Iggo, R.</dc:creator>
<dc:creator>Townley, S. L.</dc:creator>
<dc:creator>Dehairs, J.</dc:creator>
<dc:creator>Mah, C. Y.</dc:creator>
<dc:creator>Helm, M.</dc:creator>
<dc:creator>Alizadeh-Ghodsi, M.</dc:creator>
<dc:creator>Pickering, M.</dc:creator>
<dc:creator>Watt, M. J.</dc:creator>
<dc:creator>Quek, L.-E.</dc:creator>
<dc:creator>Hoy, A. J.</dc:creator>
<dc:creator>Tilley, W. D.</dc:creator>
<dc:creator>Swinnen, J. V.</dc:creator>
<dc:creator>Butler, L. M.</dc:creator>
<dc:creator>Selth, L. A.</dc:creator>
<dc:date>2022-10-14</dc:date>
<dc:identifier>doi:10.1101/2022.10.13.511039</dc:identifier>
<dc:title><![CDATA[ACSM1 and ACSM3 regulate prostate cancer fatty acid metabolism to promote tumour growth and constrain ferroptosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.13.512140v1?rss=1">
<title>
<![CDATA[
Intermittent hypoxia therapy engages multiple longevity pathways to double lifespan in C.elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.13.512140v1?rss=1</link>
<description><![CDATA[
Genetic activation of the hypoxia response robustly extends lifespan in C. elegans, while environmental hypoxia shows more limited benefit. Here we describe an intermittent hypoxia therapy (IHT) able to double the lifespan of wildtype worms. The lifespan extension observed in IHT does not require HIF-1 but is partially blocked by loss of DAF-16/FOXO. RNAseq analysis shows that IHT triggers a transcriptional state distinct from continuous hypoxia and affects down-stream genes of multiple longevity pathways. We performed a temperature sensitive forward genetic screen to isolate mutants with delayed nuclear localization of DAF-16 in response to IHT and suppression of IHT longevity. One of these mutations mapped to the enzyme Inositol Polyphosphate MultiKinase (IPMK-1). ipmk-1 mutants, like daf-16 mutants, partially suppress the benefits of IHT, while other effectors of phosphatidyl inositol signaling pathways (PLC{beta}4, IPPK, Go/i) more robustly suppress IHT longevity.

One-Sentence SummaryIntermittent hypoxia therapy is frequency dependent, HIF independent, and requires FOXO, PLC{beta}, Go/i, IPMK, and IPPK.
]]></description>
<dc:creator>Pitt, J. N.</dc:creator>
<dc:creator>Chavez, E.</dc:creator>
<dc:creator>Provencher, K. M.</dc:creator>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>Tran, C.</dc:creator>
<dc:creator>Tran, J.</dc:creator>
<dc:creator>Huang, K.</dc:creator>
<dc:creator>Vaid, A.</dc:creator>
<dc:creator>Abadir, M. L.</dc:creator>
<dc:creator>Arang, N.</dc:creator>
<dc:creator>Leiser, S. F.</dc:creator>
<dc:creator>Roth, M. B.</dc:creator>
<dc:creator>Kaeberlein, M.</dc:creator>
<dc:date>2022-10-17</dc:date>
<dc:identifier>doi:10.1101/2022.10.13.512140</dc:identifier>
<dc:title><![CDATA[Intermittent hypoxia therapy engages multiple longevity pathways to double lifespan in C.elegans]]></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/10.1101/2022.11.07.515222v1?rss=1">
<title>
<![CDATA[
Mosaic chromosomal alterations in blood across ancestries via whole-genome sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.07.515222v1?rss=1</link>
<description><![CDATA[
Mosaic mutations in blood are common with increasing age and are prognostic markers for cancer, cardiovascular dysfunction and other diseases. This group of acquired mutations include megabase-scale mosaic chromosomal alterations (mCAs). These large mutations have mainly been surveyed using SNP array data from individuals of European (EA) or Japanese genetic ancestry. To gain a better understanding of mCA rates and associated risk factors in genetically diverse populations, we surveyed whole genome sequencing data from 67,390 individuals, including 20,132 individuals of African ancestry (AA), and 7,608 of Hispanic ancestry (HA) with deep (30X) whole genome sequencing data from the NHLBI Trans Omics for Precision Medicine (TOPMed) program. We adapted an existing mCA calling algorithm for application to WGS data, and observed higher sensitivity with WGS data, compared with array-based data, in uncovering mCAs at low mutant cell fractions. As in previous reports, we observed a strong association with age and a non-uniform distribution of mCAs across the genome. The presence of autosomal (but not chromosome X) mCAs was associated with an increased risk of both lymphoid and myeloid malignancies. After adjusting for age, we found that individuals of European ancestry have the highest rates of autosomal mCAs, mirroring the higher rate of leukemia in this group. Our analysis also uncovered higher rates of chromosome X mCAs in AA and HA compared to EA, again after adjusting for age. Germline variants in ATM and MPL showed strong associations with mCAs in cis, including ancestry specific variants. And rare variant gene-burden analysis confirmed the association of putatively protein altering variants in ATM and MPL with mCAs in cis. Individual rare variants in DCPS, ADM17, PPP1R16B, and TET2 were all associated with autosomal mCAs and rare variants in OR4C16 were associated with chromosome X mCAs in females. There was significant enrichment of co-occurrence of CHIP mutations and mCAs both altering cancer associated genes TET2, DNMT3A, JAK2, CUX1, and TP53. Overall, our study demonstrates that rates of mCAs differ across populations and that rare inherited germline variants are strongly associated with mCAs across genetically diverse populations. These results strongly motivate further studies of mCAs in under-represented populations to better understand the causes and consequences of this class of somatic variation.
]]></description>
<dc:creator>Auer, P. L.</dc:creator>
<dc:creator>Jakubek, Y. A.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Stilp, A. M.</dc:creator>
<dc:creator>Bacon, J.</dc:creator>
<dc:creator>Wong, J.</dc:creator>
<dc:creator>Mitchell, B.</dc:creator>
<dc:creator>Lewis, J.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Loos, R. J. F.</dc:creator>
<dc:creator>Preuss, M.</dc:creator>
<dc:creator>Barnes, K.</dc:creator>
<dc:creator>Pankratz, N.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Redline, S.</dc:creator>
<dc:creator>Psaty, B. M.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Silverman, E.</dc:creator>
<dc:creator>Cho, M.</dc:creator>
<dc:creator>Yun, J.</dc:creator>
<dc:creator>Ramachandran, V.</dc:creator>
<dc:creator>Mathias, R. A.</dc:creator>
<dc:creator>Taub, M.</dc:creator>
<dc:creator>Peyser, P. A.</dc:creator>
<dc:creator>Smith, J. A.</dc:creator>
<dc:creator>Kardia, S.</dc:creator>
<dc:creator>Arnett, D.</dc:creator>
<dc:creator>North, K. E.</dc:creator>
<dc:creator>Raffield, L. M.</dc:creator>
<dc:creator>Carson, A.</dc:creator>
<dc:creator>Doyle, M.</dc:creator>
<dc:creator>Rich, S.</dc:creator>
<dc:creator>Rotter, J.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Cox, N.</dc:creator>
<dc:creator>Mack, T.</dc:creator>
<dc:creator>Chasman, D.</dc:creator>
<dc:creator>Machiela, M.</dc:creator>
<dc:creator>Ozcan, Z.</dc:creator>
<dc:creator>Conomos, M.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Desai, P.</dc:creator>
<dc:creator>Bick, A.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Scheet, P.</dc:creator>
<dc:date>2022-11-08</dc:date>
<dc:identifier>doi:10.1101/2022.11.07.515222</dc:identifier>
<dc:title><![CDATA[Mosaic chromosomal alterations in blood across ancestries via whole-genome sequencing]]></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/10.1101/2022.11.10.516015v1?rss=1">
<title>
<![CDATA[
Transcriptional reprogramming of natural killer cells by vaccinia virus shows both distinct and conserved features with mCMV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.10.516015v1?rss=1</link>
<description><![CDATA[
Natural killer (NK) cells have an established role in controlling poxvirus infection and there is a growing interest to exploit their capabilities in the context of poxvirus-based oncolytic therapy and vaccination. How NK cells recognise poxvirus-infected cells to become activated remains unclear. To address this knowledge gap, we studied the NK cell response to vaccinia virus (VACV) in vivo, using a systemic infection murine model. We found broad alterations in NK cells transcriptional activity in VACV-infected mice, consistent with both direct target cell recognition and cytokine exposure. There were also alterations in the expression levels of specific NK surface receptors (NKRs), including the Ly49 family and SLAM receptors, as well as upregulation of memory-associated NK markers. Despite the latter observation, adoptive transfer of NK memory populations did not confer protection from re-infection. Comparison with the NK cell response to murine cytomegalovirus (MCMV) infection highlighted common features, but also distinct NK transcriptional programmes initiated by VACV. Finally, there was a clear overlap between the NK transcriptional response in humans vaccinated with an attenuated VACV, modified vaccinia Ankara (MVA), demonstrating conservation between the NK response in these different host species. Overall, this study provides new data about NK cell activation, function, and homeostasis during VACV infection, and may have implication for the design of VACV-based therapeutics.
]]></description>
<dc:creator>Depierreux, D. M.</dc:creator>
<dc:creator>Smith, G. L.</dc:creator>
<dc:creator>Ferguson, B. J.</dc:creator>
<dc:date>2022-11-10</dc:date>
<dc:identifier>doi:10.1101/2022.11.10.516015</dc:identifier>
<dc:title><![CDATA[Transcriptional reprogramming of natural killer cells by vaccinia virus shows both distinct and conserved features with mCMV]]></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/10.1101/2022.11.19.517207v1?rss=1">
<title>
<![CDATA[
Evolution of the SARS-CoV-2 mutational spectrum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.19.517207v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 evolves rapidly in part because of its high mutation rate. Here we examine whether this mutational process itself has changed during viral evolution. To do this, we quantify the relative rates of different types of single nucleotide mutations at four-fold degenerate sites in the viral genome across millions of human SARS-CoV-2 sequences. We find clear shifts in the relative rates of several types of mutations during SARS-CoV-2 evolution. The most striking trend is a roughly two-fold decrease in the relative rate of G[-&gt;]T mutations in Omicron versus early clades, as was recently noted by Ruis et al (2022). There is also a decrease in the relative rate of C[-&gt;]T mutations in Delta, and other subtle changes in the mutation spectrum along the phylogeny. We speculate that these changes in the mutation spectrum could arise from viral mutations that affect genome replication, packaging, and antagonization of host innate-immune factors--although environmental factors could also play a role. Interestingly, the mutation spectrum of Omicron is more similar than that of earlier SARS-CoV-2 clades to the spectrum that shaped the long-term evolution of sarbecoviruses. Overall, our work shows that the mutation process is itself a dynamic variable during SARS-CoV-2 evolution, and suggests that human SARS-CoV-2 may be trending towards a mutation spectrum more similar to that of other animal sarbecoviruses.
]]></description>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Beichman, A. C.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:creator>Harris, K.</dc:creator>
<dc:date>2022-11-21</dc:date>
<dc:identifier>doi:10.1101/2022.11.19.517207</dc:identifier>
<dc:title><![CDATA[Evolution of the SARS-CoV-2 mutational spectrum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.05.519107v1?rss=1">
<title>
<![CDATA[
Autorepression-based conditional gene expression system in yeast for variation-suppressed control of protein dosage 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.05.519107v1?rss=1</link>
<description><![CDATA[
Conditional control of gene expression allows an experimenter to investigate many aspects of a genes function. In the model organism Saccharomyces cerevisiae, a number of methods to control gene expression are widely practiced, including induction by metabolites, small molecules, and even light. However, all current methods suffer from at least one of a set of drawbacks, including need for specialized growth conditions, leaky expression, or the requirement of specialized equipment. Here we describe protocols using two transformations to construct strains that carry a new controller, in which all these drawbacks are overcome. In these strains, the expression of a controlled gene (gene of interest, or GOI) is repressed by the bacterial repressor TetR, and induced by anhydrotetracycline. TetR also regulates its own expression, creating an autorepression loop. This autorepression allows tight control of gene expression/ protein dosage with low cell to cell variation in expression. A second repressor, TetR-Tup1, prevents any leaky expression. We also present a protocol showing a particular workhorse application of such strains, to generate synchronized cell populations. We turn off the expression of the cell cycle regulator CDC20 completely, arresting the cell population, and then back on so that the synchronized cells resume cell cycle progression. This control system can be applied to any endogenous or exogenous gene for precise expression.

Basic Protocol 1Generating a parent WTC846 strain.

Basic Protocol 2Generating a WTC846 strain with controlled expression of the targeted gene

Alternate Protocol 1CRISPR-mediated promoter replacement

Basic Protocol 3Cell cycle synchronization/Arrest and Release using the WTC846-K3::CDC20 strain
]]></description>
<dc:creator>Azizoglu, A.</dc:creator>
<dc:creator>Loureiro, C.</dc:creator>
<dc:creator>Venetz, J.</dc:creator>
<dc:creator>Brent, R.</dc:creator>
<dc:date>2022-12-08</dc:date>
<dc:identifier>doi:10.1101/2022.12.05.519107</dc:identifier>
<dc:title><![CDATA[Autorepression-based conditional gene expression system in yeast for variation-suppressed control of protein dosage]]></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/10.1101/2022.12.09.519706v1?rss=1">
<title>
<![CDATA[
Distinct immune responses associated with vaccination status and protection outcomes after malaria challenge 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.09.519706v1?rss=1</link>
<description><![CDATA[
Understanding immune mechanisms that mediate malaria protection is critical for improving vaccine development. Vaccination with radiation-attenuated Plasmodium falciparum sporozoites (PfRAS) induces high level of sterilizing immunity against malaria and serves as a valuable tool for the study of protective mechanisms. To identify vaccine-induced and protection-associated responses during malaria infection, we performed transcriptome profiling of whole blood and in-depth cellular profiling of PBMCs from volunteers who received either PfRAS or noninfectious mosquito bites, followed by controlled human malaria infection (CHMI) challenge. In-depth single-cell profiling of cell subsets that respond to CHMI in mock-vaccinated individuals showed a predominantly inflammatory transcriptome response. Whole blood transcriptome analysis revealed that gene sets associated with interferon responses and T and B cell signatures were increased and decreased, respectively, in protected vaccinees as early as one day following CHMI. In contrast, non-protected vaccinees and mock-vaccinated individuals exhibited shared transcriptome changes after CHMI characterized by decreased innate cell signatures and inflammatory responses. Additionally, immunophenotyping data showed different induction profiles of v{delta}2+{gamma}{delta} T cells, CD56+ CD8+ T effector memory (Tem) cells, and non-classical monocytes between protected vaccinees and individuals developing blood-stage parasitemia, following treatment and resolution of infection. Our data provide key insights in understanding immune mechanistic pathways of PfRAS-induced protection and infective CHMI. We demonstrate that protective immunity by PfRAS is associated with early changes in interferon and adaptive immune responses.

Author summaryMalaria poses a significant global health threat, causing over half a million deaths annually. Effective vaccines are critically needed to prevent malaria disease. Our incomplete understanding of immune mechanisms that mediate malaria protection is hampering the development of effective vaccines. Irradiated sporozoite vaccines can induce highly sterilizing protection against malaria and are a valuable tool for the analysis of immune protection. Here, we aimed to characterize correlates of immune protection in individuals vaccinated with a suboptimal dose of irradiated sporozoite and subsequently challenged with live malaria parasite. Blood samples were taken before and after malaria challenge, and gene expression and cell type profiles were measured. We observed that the trajectories of immune response after malaria challenge is highly distinct between protected and non-protected vaccinees. We observed early perturbations in interferon response and adaptive immune cells in protected vaccinees whereas inflammatory and innate cell response were unique to non-protected vaccinees. We also observed that the immune profile after malaria challenge was distinctly similar between non-protected vaccinees and mock-vaccinated individuals. Our study sheds light on the dynamics of vaccine-induced immune responses that are associated with protection from malaria after CHMI.
]]></description>
<dc:creator>Oyong, D. A.</dc:creator>
<dc:creator>Duffy, F. J.</dc:creator>
<dc:creator>Neal, M. L.</dc:creator>
<dc:creator>Du, Y.</dc:creator>
<dc:creator>Carnes, J.</dc:creator>
<dc:creator>Schwedhelm, K. V.</dc:creator>
<dc:creator>Hertoghs, N.</dc:creator>
<dc:creator>Jun, S.-H.</dc:creator>
<dc:creator>Miller, H.</dc:creator>
<dc:creator>Aitchison, J. D.</dc:creator>
<dc:creator>De Rosa, S. C.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>McDermott, S. M.</dc:creator>
<dc:creator>Stuart, K. D.</dc:creator>
<dc:date>2022-12-10</dc:date>
<dc:identifier>doi:10.1101/2022.12.09.519706</dc:identifier>
<dc:title><![CDATA[Distinct immune responses associated with vaccination status and protection outcomes after malaria challenge]]></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/10.1101/2022.12.21.521494v1?rss=1">
<title>
<![CDATA[
Sex-biased Yap1 oncogene function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.21.521494v1?rss=1</link>
<description><![CDATA[
The incidence of many human cancers differs according to sex, but little is known about the interplay between oncogenic events and sex as a variable in tumorigenesis. Here we report that the oncogene Yap1 is sexually dimorphic in medulloblastoma progression and immune suppression. We show that Yap1 promotes stemness and blocks differentiation in sonic hedgehog (SHH)-subtype medulloblastoma by at least two distinct but complementary molecular mechanisms to regulate the RNA expression and protein functions of Sox2, Atoh1, NeuroD1, and Zic1/2. Yap1 also promotes an immune suppressive tumor microenvironment by directly regulating Csf1, Igf1, and Igfbp3 transcription and modulating IL6-JAK-STAT3, TNFR1, TGF-{beta}, and CCL5 immune pathways. Notably, Yap1 function is more critical in males and this is evolutionarily conserved: genes downstream of YAP1 identified in mouse models stratify male but not female medulloblastoma patient survival. In summary, we demonstrate a sex-based function for an oncogene, underscoring the critical need to incorporate sex as a variable in cancer mechanism and clinical response studies, particularly those involving YAP1.
]]></description>
<dc:creator>Abdelfattah, N.</dc:creator>
<dc:creator>Natarajan, S.</dc:creator>
<dc:creator>Maldonado, J.</dc:creator>
<dc:creator>Tran, H. N.</dc:creator>
<dc:creator>McMinimy, R.</dc:creator>
<dc:creator>Borland, H.</dc:creator>
<dc:creator>Chen, S.-h.</dc:creator>
<dc:creator>Camargo, F.</dc:creator>
<dc:creator>Olson, J.</dc:creator>
<dc:creator>George, J.</dc:creator>
<dc:creator>Yun, K.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.21.521494</dc:identifier>
<dc:title><![CDATA[Sex-biased Yap1 oncogene function]]></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/10.1101/2023.01.06.522784v1?rss=1">
<title>
<![CDATA[
Antigen-dependent IL-12 signaling in CAR T cells promotes regional to systemic disease targeting 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.06.522784v1?rss=1</link>
<description><![CDATA[
Chimeric antigen receptor (CAR) T cell therapeutic responses are hampered by limited T cell trafficking, persistence, and durable anti-tumor activity in solid tumor microenvironments. However, these challenges can be largely overcome by relatively unconstrained synthetic engineering strategies, which are being harnessed to improve solid tumor CAR T cell therapies. Here, we describe fully optimized CAR T cells targeting tumor-associated glycoprotein-72 (TAG72) for the treatment of solid tumors, identifying the CD28 transmembrane domain upstream of the 4-1BB co-stimulatory domain as a driver of potent anti-tumor activity and IFN{gamma} secretion. These findings have culminated into a phase 1 trial evaluating safety, feasibility, and bioactivity of TAG72-CAR T cells for the treatment of patients with advanced ovarian cancer (NCT05225363). Preclinically, we found that CAR T cell-mediated IFN{gamma} production facilitated by IL-12 signaling was required for tumor cell killing, which was recapitulated by expressing an optimized membrane-bound IL-12 (mbIL12) molecule on CAR T cells. Critically, mbIL12 cell surface expression and downstream signaling was induced and sustained only following CAR T cell activation. CAR T cells with mbIL12 demonstrated improved antigen-dependent T cell proliferation and potent cytotoxicity in recursive tumor cell killing assays in vitro and showed robust in vivo anti-tumor efficacy in human xenograft models of ovarian cancer peritoneal metastasis. Further, locoregional administration of TAG72-CAR T cells with antigen-dependent IL-12 signaling promoted durable anti-tumor responses against both regional and systemic disease in mice and was associated with improved systemic T cell persistence. Our study features a clinically-applicable strategy to improve the overall efficacy of locoregionally-delivered CAR T cells engineered with antigen-dependent immune-modulating cytokines in targeting both regional and systemic disease.
]]></description>
<dc:creator>Lee, E.</dc:creator>
<dc:creator>Cullen, C.</dc:creator>
<dc:creator>Murad, J. P.</dc:creator>
<dc:creator>Gumber, D.</dc:creator>
<dc:creator>Park, A.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Stern, L.</dc:creator>
<dc:creator>Adkins, L.</dc:creator>
<dc:creator>Dhapola, G.</dc:creator>
<dc:creator>Gittins, B.</dc:creator>
<dc:creator>Chang, W.-C.</dc:creator>
<dc:creator>Martinez, C.</dc:creator>
<dc:creator>Woo, Y.</dc:creator>
<dc:creator>Cristea, M.</dc:creator>
<dc:creator>Rodriguez, L.</dc:creator>
<dc:creator>Ishihara, J.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:creator>Forman, S. J.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Priceman, S. J.</dc:creator>
<dc:date>2023-01-07</dc:date>
<dc:identifier>doi:10.1101/2023.01.06.522784</dc:identifier>
<dc:title><![CDATA[Antigen-dependent IL-12 signaling in CAR T cells promotes regional to systemic disease targeting]]></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/10.1101/2023.01.17.524235v1?rss=1">
<title>
<![CDATA[
RepairSwitch: simultaneous functional assessment of homologous recombination and end joining DNA repair pathways in living cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.17.524235v1?rss=1</link>
<description><![CDATA[
DNA repair pathways are frequently defective in human cancers. DNA double strand breaks (DSBs) are most often repaired by either homologous recombination (HR) or non-homologous end joining (NHEJ). Alterations in repair pathways can indicate sensitivity to therapeutic agents such as PARP inhibitors, cisplatin, and immunotherapy. Thus, functional assays to measure rates of HR and NHEJ are of significant interest. Several methods have been developed to measure rates of HR or NHEJ; however, there is a need for functional cell-based assays that can measure rates by both major DNA DSB pathways simultaneously. Here, we describe the RepairSwitch assay, a flow cytometry assay to assess rates of HR and NHEJ mediated repair of Cas9 programmed DSB simultaneously using a novel fluorescence switching reporter system. The assay exhibits low background signal and is capable of detecting rare repair events in the 1 in 10,000 range. We demonstrate the utility of RepairSwitch by measuring the potency of inhibitors of ATM (KU-60019, KU-55933), DNA-PK (NU7441), and PARP (Olaparib) on modulating DSB repair rates in HEK293FT cells. The selective ATM inhibitor KU-60019 inhibited HR rates with IC50 of 915 nM. Interestingly, KU-60019 exposure led to a dose responsive increase in rates of NHEJ. In contrast, the less selective ATM inhibitor KU-55933, which also has activity on DNA-PK, showed inhibition of both HR and NHEJ. The selective DNA-PK inhibitor NU7441 inhibited NHEJ efficiency with an IC50 of 299 nM, and showed a dose responsive increase in HR. The PARP inhibitor Olaparib showed lower potency in modulating HR and NHEJ. We next used the RepairSwitch assay to assess how pharmacological and genetic inhibition of DNA methyltransferases (DNMT) impacted rates of HR and NHEJ. The DNMT inhibitor decitabine reduced HR, but increased rates of NHEJ, both in a dose responsive manner, in both HEK293FT and HCT116 cells (IC50 for HR of 187 nM and 1.4 uM respectively). Knockout of DNMT1 and DNMT3B increased NHEJ, while knockout of DNMT3B, but not DNMT1, reduced HR. These results illustrate the utility of RepairSwitch as a functional assay for measuring changes in rates of DSB repair induced by pharmacological or genetic perturbation. Furthermore, the findings illustrate the potential for one DNA repair mechanism to compensate in part for loss of another. Finally, we showed that inhibition of DNMT can lead to reduction of HR and increase in NHEJ, providing some additional insight into recently observed synergy of DNMT inhibitors with PARP inhibitors for cancer treatment.
]]></description>
<dc:creator>Steinberg, R. C.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Vaghasia, A. M.</dc:creator>
<dc:creator>Giovinazzo, H.</dc:creator>
<dc:creator>Pham, M.-T. N.</dc:creator>
<dc:creator>Tselenchuk, D.</dc:creator>
<dc:creator>Chikarmane, R.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Nelson, W. G.</dc:creator>
<dc:creator>Yegnasubramanian, S.</dc:creator>
<dc:date>2023-01-17</dc:date>
<dc:identifier>doi:10.1101/2023.01.17.524235</dc:identifier>
<dc:title><![CDATA[RepairSwitch: simultaneous functional assessment of homologous recombination and end joining DNA repair pathways in living cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-17</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">
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]]></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/10.1101/2023.01.25.525495v1?rss=1">
<title>
<![CDATA[
A Distinct Phenotype of Polarized Memory B cell holds IgE Memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.25.525495v1?rss=1</link>
<description><![CDATA[
Allergen-specific IgE antibodies mediate allergic pathology in diseases such as allergic rhinitis and food allergy. Memory B cells (MBCs) contribute to circulating IgE by regenerating IgE-producing plasma cells upon allergen encounter. We report a population of type 2 polarized MBCs defined as CD23hi, IL-4Rhi, CD32low at the transcriptional and surface protein levels. These "MBC2s" are enriched in IgG1 and IgG4-expressing cells, while constitutively expressing germline transcripts for IgE. Allergen-specific B cells from patients with allergic rhinitis and food allergy were enriched in MBC2s. MBC2s generated allergen specific-IgE during sublingual immunotherapy, thereby identifying these cells as the primary reservoir of IgE. The identification of MBC2s provides insights into the maintenance of IgE memory, which is detrimental in allergic diseases, but which could be beneficial in protection against venoms and helminths.

One-Sentence SummaryIdentification of a novel memory B cell subset which holds allergen specific IgE memory.
]]></description>
<dc:creator>Koenig, J. F.</dc:creator>
<dc:creator>Knudsen, N. P. H.</dc:creator>
<dc:creator>Phelps, A.</dc:creator>
<dc:creator>Bruton, K.</dc:creator>
<dc:creator>Hoof, I.</dc:creator>
<dc:creator>Lund, G.</dc:creator>
<dc:creator>Libera, D. D.</dc:creator>
<dc:creator>Lund, A.</dc:creator>
<dc:creator>Christensen, L.</dc:creator>
<dc:creator>Glass, D. R.</dc:creator>
<dc:creator>Walker, T. D.</dc:creator>
<dc:creator>Fang, A.</dc:creator>
<dc:creator>Waserman, S.</dc:creator>
<dc:creator>Jordana, M.</dc:creator>
<dc:creator>Andersen, P. S.</dc:creator>
<dc:date>2023-01-25</dc:date>
<dc:identifier>doi:10.1101/2023.01.25.525495</dc:identifier>
<dc:title><![CDATA[A Distinct Phenotype of Polarized Memory B cell holds IgE Memory]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.28.526014v1?rss=1">
<title>
<![CDATA[
Myeloperoxidase promotes a tumorigenic microenvironment in non-small cell lungcancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.28.526014v1?rss=1</link>
<description><![CDATA[
Myeloperoxidase (MPO) is a heme peroxidase that is mainly expressed and secreted by neutrophils. MPOs role in inflammatory diseases has been highlighted in recent years, but its role in tumor development remains unclear. Therefore, we investigated the role of MPO in non-small cell lung cancer (NSCLC). In silico analysis revealed a survival benefit in patients with NSCLC and low MPO expression. Furthermore, a syngeneic tumor model using MPO knockout (KO) mice revealed that mice lacking MPO had lower tumor growth than controls. The reduction in tumor size was accompanied by an increase in lymphoid populations, including natural killer cells and CD8+ T cells, suggesting a shift to a more anti-tumorigenic immune environment in MPO-KO mouse tumors. The T cell induced interferon-gamma (IFN-{gamma}) expression was increased in MPO-KO tumors, indicating increased tumoricidal activity. CD8 depletion abolished the previously observed reduction in tumor size in MPO-KO mice, indicating that CD8+ T cells play an important role. In vitro, T cells treated with MPO showed reduced proliferation and IFN-{gamma} expression. Furthermore, MPO could be internalized into T cells. Heparin pretreatment of T cells blocked MPO binding and internalization into T cells and reversed MPO-induced proliferation reduction. Interestingly, MPO+ lymphocytes were found in tumor samples from patients with NSCLC. Our findings suggest that MPO plays an immunosuppressive role in NSCLC.

One Sentence SummaryHigh myeloperoxidase (MPO) expression in non-small cell lung cancer patients is a predictor for adverse outcome and mice lacking MPO showed enhanced anti-tumorigenic leukocyte infiltration, suggesting a pro-tumorigenic role of MPO.
]]></description>
<dc:creator>Valadez-Cosmes, P.</dc:creator>
<dc:creator>Kathrin, M.</dc:creator>
<dc:creator>Kindler, O.</dc:creator>
<dc:creator>Cosic-Mujkanovic, N.</dc:creator>
<dc:creator>Lueger, A.</dc:creator>
<dc:creator>Raftopoulou, S.</dc:creator>
<dc:creator>Kienzl, M.</dc:creator>
<dc:creator>Mihalic, Z. N.</dc:creator>
<dc:creator>Santiso, A.</dc:creator>
<dc:creator>Sarsembayeva, A.</dc:creator>
<dc:creator>Brcic, L.</dc:creator>
<dc:creator>Lindenmann, J.</dc:creator>
<dc:creator>Sattler, W.</dc:creator>
<dc:creator>Heinemann, A.</dc:creator>
<dc:creator>Schicho, R.</dc:creator>
<dc:creator>Marsche, G.</dc:creator>
<dc:creator>Houghton, A. M.</dc:creator>
<dc:creator>Kargl, J.</dc:creator>
<dc:date>2023-01-29</dc:date>
<dc:identifier>doi:10.1101/2023.01.28.526014</dc:identifier>
<dc:title><![CDATA[Myeloperoxidase promotes a tumorigenic microenvironment in non-small cell lungcancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.30.526314v1?rss=1">
<title>
<![CDATA[
Fitness effects of mutations to SARS-CoV-2 proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.30.526314v1?rss=1</link>
<description><![CDATA[
Knowledge of the fitness effects of mutations to SARS-CoV-2 can inform assessment of new variants, design of therapeutics resistant to escape, and understanding of the functions of viral proteins. However, experimentally measuring effects of mutations is challenging: we lack tractable lab assays for many SARS-CoV-2 proteins, and comprehensive deep mutational scanning has been applied to only two SARS-CoV-2 proteins. Here we develop an approach that leverages millions of publicly available SARS-CoV-2 sequences to estimate effects of mutations. We first calculate how many independent occurrences of each mutation are expected to be observed along the SARS-CoV-2 phylogeny in the absence of selection. We then compare these expected observations to the actual observations to estimate the effect of each mutation. These estimates correlate well with deep mutational scanning measurements. For most genes, synonymous mutations are nearly neutral, stop-codon mutations are deleterious, and amino-acid mutations have a range of effects. However, some viral accessory proteins are under little to no selection. We provide interactive visualizations of effects of mutations to all SARS-CoV-2 proteins (https://jbloomlab.github.io/SARS2-mut-fitness/). The framework we describe is applicable to any virus for which the number of available sequences is sufficiently large that many independent occurrences of each neutral mutation are observed.
]]></description>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:date>2023-01-31</dc:date>
<dc:identifier>doi:10.1101/2023.01.30.526314</dc:identifier>
<dc:title><![CDATA[Fitness effects of mutations to SARS-CoV-2 proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.30.526360v1?rss=1">
<title>
<![CDATA[
A comprehensive rRNA variation atlas in health and disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.30.526360v1?rss=1</link>
<description><![CDATA[
Ribosomal DNA and RNA (rDNA and rRNA) sequences are usually discarded from sequencing analyses. But with hundreds of copies of rDNA genes it is unknown whether they possess sequence variations that form different types of ribosomes that affect human physiology and disease. Here, we developed an algorithm for variant-calling between paralog genes (termed RGA) and compared rDNA variations found in short- and long-read sequencing data from the 1,000 Genomes Project (1KGP) and Genome In A Bottle (GIAB). We additionally developed a novel protocol for long-read sequencing full-length rRNA (RIBO-RT) from actively translating ribosomes. Our analyses identified hundreds of rDNA variants, most of which, surprisingly, are short insertion-deletions (indels) and dozens of highly abundant rRNA variants that are incorporated into translationally active ribosomes. To visualize variant ribosomes at the single cell level, we developed an in-situ rRNA sequencing method (SWITCH-seq) which revealed that variants are co-expressed within individual cells. Strikingly, by analyzing rDNA, we found that variants assemble into distinct ribosome subtypes. We discovered that these subtypes acquire different rRNA structures by successfully employing dimethyl sulfate (DMS) probing of full length rRNA. With this atlas we investigated rRNA variation changes across human tissues and cancer types. This revealed tissue-specific rRNA subtype expression in endoderm/ectoderm-derived tissues. In cancer, low abundant rRNA variants can become highly expressed, which suggests the presence of cancer-specific ribosomes. Together, this study identifies and comprehensively characterizes the diversity of ribosomes at the level of rRNA variants which is dominated by indel variants, their chromosomal location and unique structure as well as the association of ribosome variation with tissue-specific biology and cancer.
]]></description>
<dc:creator>Rothschild, D.</dc:creator>
<dc:creator>Susanto, T. T.</dc:creator>
<dc:creator>Spence, J. P.</dc:creator>
<dc:creator>Genuth, N. R.</dc:creator>
<dc:creator>Sinnott-Armstrong, N.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:creator>Barna, M.</dc:creator>
<dc:date>2023-02-02</dc:date>
<dc:identifier>doi:10.1101/2023.01.30.526360</dc:identifier>
<dc:title><![CDATA[A comprehensive rRNA variation atlas in health and disease]]></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/10.1101/2023.01.31.526505v1?rss=1">
<title>
<![CDATA[
Genetic control of mRNA splicing as a potential mechanism for incomplete penetrance of rare coding variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.31.526505v1?rss=1</link>
<description><![CDATA[
Exonic variants present some of the strongest links between genotype and phenotype. However, these variants can have significant inter-individual pathogenicity differences, known as variable penetrance. In this study, we propose a model where genetically controlled mRNA splicing modulates the pathogenicity of exonic variants. By first cataloging exonic inclusion from RNA-seq data in GTEx v8, we find that pathogenic alleles are depleted on highly included exons. Using a large-scale phased WGS data from the TOPMed consortium, we observe that this effect may be driven by common splice-regulatory genetic variants, and that natural selection acts on haplotype configurations that reduce the transcript inclusion of putatively pathogenic variants, especially when limiting to haploinsufficient genes. Finally, we test if this effect may be relevant for autism risk using families from the Simons Simplex Collection, but find that splicing of pathogenic alleles has a penetrance reducing effect here as well. Overall, our results indicate that common splice-regulatory variants may play a role in reducing the damaging effects of rare exonic variants.
]]></description>
<dc:creator>Einson, J.</dc:creator>
<dc:creator>Glinos, D.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Castaldi, P.</dc:creator>
<dc:creator>Darbar, D.</dc:creator>
<dc:creator>de Andrade, M.</dc:creator>
<dc:creator>Ellinor, P.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Gabriel, S.</dc:creator>
<dc:creator>Germer, S.</dc:creator>
<dc:creator>Gibbs, R.</dc:creator>
<dc:creator>Hersh, C.</dc:creator>
<dc:creator>Johnsen, J.</dc:creator>
<dc:creator>Kaplan, R.</dc:creator>
<dc:creator>Konkle, B.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Nassir, R.</dc:creator>
<dc:creator>Loos, R. J. F.</dc:creator>
<dc:creator>Meyers, D. A.</dc:creator>
<dc:creator>Mitchell, B. D.</dc:creator>
<dc:creator>Psaty, B.</dc:creator>
<dc:creator>Vasan, R. S.</dc:creator>
<dc:creator>Rich, S. S.</dc:creator>
<dc:creator>Rienstra, M.</dc:creator>
<dc:creator>Rotter, J. I.</dc:creator>
<dc:creator>Saferali, A.</dc:creator>
<dc:creator>Shoemaker, M. B.</dc:creator>
<dc:creator>Silverman, E.</dc:creator>
<dc:creator>Smith, A. V.</dc:creator>
<dc:creator>Mohammadi, P.</dc:creator>
<dc:creator>Castel, S. E.</dc:creator>
<dc:creator>Iossifov, I.</dc:creator>
<dc:creator>Lappalainen, T.</dc:creator>
<dc:date>2023-01-31</dc:date>
<dc:identifier>doi:10.1101/2023.01.31.526505</dc:identifier>
<dc:title><![CDATA[Genetic control of mRNA splicing as a potential mechanism for incomplete penetrance of rare coding variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.03.527038v1?rss=1">
<title>
<![CDATA[
BAF chromatin complexes do not mediate GLI transcriptional repression of Hedgehog target genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.03.527038v1?rss=1</link>
<description><![CDATA[
Transcriptional responses to the Hedgehog (HH) signaling pathway are primarily modulated by GLI repression in the mouse limb. Previous studies suggested a role for the BAF chromatin remodeling complex in mediating GLI repression. Consistent with this possibility, the core BAF complex protein SMARCC1 is present at most active limb enhancers including the majority of GLI enhancers. However, in contrast to GLI repression which reduces chromatin accessibility, SMARCC1 maintains chromatin accessibility at most enhancers, including those bound by GLI. Moreover, SMARCC1 binding at GLI-regulated enhancers occurs independently of GLI3. Consistent with previous studies, some individual GLI target genes are mis-regulated in Smarcc1 conditional knockouts, though most GLI target genes are unaffected. Moreover, SMARCC1 is not necessary for mediating constitutive GLI repression in HH mutant limb buds. We conclude that SMARCC1 does not mediate GLI3 repression, which we propose utilizes alternative chromatin remodeling complexes.
]]></description>
<dc:creator>Ramachandran, J.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Lex, R. K.</dc:creator>
<dc:creator>Windsor, K.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Ji, H.</dc:creator>
<dc:creator>Vokes, S. A.</dc:creator>
<dc:date>2023-02-08</dc:date>
<dc:identifier>doi:10.1101/2023.02.03.527038</dc:identifier>
<dc:title><![CDATA[BAF chromatin complexes do not mediate GLI transcriptional repression of Hedgehog target genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.02.530850v1?rss=1">
<title>
<![CDATA[
Type I interferon dynamics determines memory B cell epigenetic identity in chronic viral infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.02.530850v1?rss=1</link>
<description><![CDATA[
Memory B cells are key providers of long-lived immunity against infectious disease, yet in chronic viral infection they do not produce effective protection. How chronic viral infection disrupts memory B cell development, and whether such changes are reversible, remains unknown. Here, we uncover type-I interferon (IFN-I) dynamics as a key determinant in shaping chronic memory B cell development. Through single-cell (sc)ATAC-sequencing and scRNA-sequencing, we identified a unique memory subset enriched for IFN-stimulated genes (ISGs) during chronic lymphocytic choriomeningitis virus infection. Blockade of IFNAR-1 early in infection transformed the chromatin landscape of chronic memory B cells, decreasing accessibility at ISG-inducing transcription factor binding motifs and inducing a phenotypic change in the dominating memory B cell subset. However, timing was critical, with memory B cells resistant to intervention after 4 weeks post-infection. Together, our research identifies a key mechanism to instruct memory B cell identity during viral infection.

One Sentence Summary: IFN dynamics in chronic versus acute viral infection determines memory B cell development.
]]></description>
<dc:creator>Cooper, L.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Polmear, J.</dc:creator>
<dc:creator>Szeto, C.</dc:creator>
<dc:creator>Pang, E. S.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:creator>Jackson, K. J. L.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>La Gruta, N.</dc:creator>
<dc:creator>Martelotto, L. G.</dc:creator>
<dc:creator>Parish, I. A.</dc:creator>
<dc:creator>O'Keeffe, M.</dc:creator>
<dc:creator>Scharer, C. D.</dc:creator>
<dc:creator>Gras, S.</dc:creator>
<dc:creator>Good-Jacobson, K. L.</dc:creator>
<dc:date>2023-03-02</dc:date>
<dc:identifier>doi:10.1101/2023.03.02.530850</dc:identifier>
<dc:title><![CDATA[Type I interferon dynamics determines memory B cell epigenetic identity in chronic viral infection]]></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/10.1101/2023.03.05.531201v1?rss=1">
<title>
<![CDATA[
High-Capacity Sample Multiplexing for Single Cell Chromatin Accessibility Profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.05.531201v1?rss=1</link>
<description><![CDATA[
Single-cell chromatin accessibility has emerged as a powerful means of understanding the epigenetic landscape of diverse tissues and cell types, but profiling cells from many independent specimens is challenging and costly. Here we describe a novel approach, sciPlex-ATAC-seq, which uses unmodified DNA oligos as sample-specific nuclear labels, enabling the concurrent profiling of chromatin accessibility within single nuclei from virtually unlimited specimens or experimental conditions. We first demonstrate our method with a chemical epigenomics screen, in which we identify drug-altered distal regulatory sites predictive of compound- and dose-dependent effects on transcription. We then analyze cell type-specific chromatin changes in PBMCs from multiple donors responding to synthetic and allogeneic immune stimulation. We quantify stimulation-altered immune cell compositions and isolate the unique effects of allogeneic stimulation on chromatin accessibility specific to T-lymphocytes. Finally, we observe that impaired global chromatin decondensation often coincides with chemical inhibition of allogeneic T-cell activation.
]]></description>
<dc:creator>Booth, G. T.</dc:creator>
<dc:creator>Daza, R. M.</dc:creator>
<dc:creator>Srivatsan, S. R.</dc:creator>
<dc:creator>McFaline-Figueroa, J. L.</dc:creator>
<dc:creator>Green Gladden, R.</dc:creator>
<dc:creator>Furlan, S. N.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:date>2023-03-06</dc:date>
<dc:identifier>doi:10.1101/2023.03.05.531201</dc:identifier>
<dc:title><![CDATA[High-Capacity Sample Multiplexing for Single Cell Chromatin Accessibility Profiling]]></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/10.1101/2023.03.13.532438v1?rss=1">
<title>
<![CDATA[
Leukemia core transcriptional circuitry is a sparsely interconnected hierarchy stabilized by incoherent feed-forward loops 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.13.532438v1?rss=1</link>
<description><![CDATA[
Lineage-defining transcription factors form densely interconnected circuits in chromatin occupancy assays, but the functional significance of these networks remains underexplored. We reconstructed the functional topology of a leukemia cell transcription network from the direct gene-regulatory programs of eight core transcriptional regulators established in pre-steady state assays coupling targeted protein degradation with nascent transcriptomics. The core regulators displayed narrow, largely non-overlapping direct transcriptional programs, forming a sparsely interconnected functional hierarchy stabilized by incoherent feed-forward loops. BET bromodomain and CDK7 inhibitors disrupted the core regulators direct programs, acting as mixed agonists/antagonists. The network is predictive of dynamic gene expression behaviors in time-resolved assays and clinically relevant pathway activity in patient populations.
]]></description>
<dc:creator>Harada, T.</dc:creator>
<dc:creator>Kalfon, J.</dc:creator>
<dc:creator>Perez, M. W.</dc:creator>
<dc:creator>Eagle, K.</dc:creator>
<dc:creator>Braes, F. D.</dc:creator>
<dc:creator>Batley, R.</dc:creator>
<dc:creator>Heshmati, Y.</dc:creator>
<dc:creator>Ferrucio, J. X.</dc:creator>
<dc:creator>Ewers, J.</dc:creator>
<dc:creator>Mehta, S.</dc:creator>
<dc:creator>Kossenkov, A.</dc:creator>
<dc:creator>Ellegast, J. M.</dc:creator>
<dc:creator>Bowker, A.</dc:creator>
<dc:creator>Wickramasinghe, J.</dc:creator>
<dc:creator>Nabet, B.</dc:creator>
<dc:creator>Paralkar, V. R.</dc:creator>
<dc:creator>Dharia, N. V.</dc:creator>
<dc:creator>Stegmaier, K.</dc:creator>
<dc:creator>Orkin, S. H.</dc:creator>
<dc:creator>Pimkin, M.</dc:creator>
<dc:date>2023-03-15</dc:date>
<dc:identifier>doi:10.1101/2023.03.13.532438</dc:identifier>
<dc:title><![CDATA[Leukemia core transcriptional circuitry is a sparsely interconnected hierarchy stabilized by incoherent feed-forward loops]]></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/10.1101/2023.03.19.533358v1?rss=1">
<title>
<![CDATA[
Pro-inflammatory feedback loops define immune responses to pathogenic lentivirus infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.19.533358v1?rss=1</link>
<description><![CDATA[
HIV causes chronic inflammation and AIDS in humans, though the rate of disease progression varies between individuals. Similarly, simian lentiviruses vary in their pathogenicity based on characteristics of both the host (simian species) and virus strain. Here, we profile immune responses in pig-tailed macaques infected with variants of SIV that differ in virulence to understand the immune mechanisms underlying lentiviral pathogenicity. Compared to a minimally pathogenic lentiviral variant, infection with a highly pathogenic variant results in a more delayed, broad, and sustained activation of inflammatory pathways, including an extensive global interferon signature. Conversely, individual cells infected with highly pathogenic lentivirus upregulated fewer interferon-stimulated genes at a lower magnitude, indicating that highly pathogenic lentivirus has evolved to partially escape from interferon responses. Further, we identified distinct gene co-expression patterns and cell-cell communication pathways that implicate CXCL10 and CXCL16 as important molecular drivers of inflammatory pathways specifically in response to highly pathogenic lentivirus infection. Immune responses to highly pathogenic lentivirus infection are characterized by amplifying regulatory circuits of pro-inflammatory cytokines with dense longitudinal connectivity. Our work presents a model of lentiviral pathogenicity where failures in early viral control mechanisms lead to delayed, sustained, and amplifying pro-inflammatory circuits, which has implications for other viral infections with highly variable disease courses.
]]></description>
<dc:creator>Wilk, A. J.</dc:creator>
<dc:creator>Marceau, J. O.</dc:creator>
<dc:creator>Kazer, S. W.</dc:creator>
<dc:creator>Fleming, I.</dc:creator>
<dc:creator>Miao, V. N.</dc:creator>
<dc:creator>Galvez-Reyes, J.</dc:creator>
<dc:creator>Shalek, A. K.</dc:creator>
<dc:creator>Holmes, S.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Blish, C. A.</dc:creator>
<dc:date>2023-03-19</dc:date>
<dc:identifier>doi:10.1101/2023.03.19.533358</dc:identifier>
<dc:title><![CDATA[Pro-inflammatory feedback loops define immune responses to pathogenic lentivirus infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.21.533528v1?rss=1">
<title>
<![CDATA[
HIV-1 Vpr combats the PU.1-driven antiviral response in primary human macrophages. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.21.533528v1?rss=1</link>
<description><![CDATA[
HIV-1 Vpr promotes efficient spread of HIV-1 from macrophages to T cells by transcriptionally downmodulating restriction factors that target HIV-1 Envelope protein (Env). Here we find that Vpr induces broad transcriptomic changes by targeting PU.1, a transcription factor necessary for expression of host innate immune response genes, including those that target Env. Consistent with this, we find silencing PU.1 in infected macrophages lacking Vpr rescues Env. Vpr downmodulates PU.1 through a proteasomal degradation pathway that depends on physical interactions with PU.1 and DCAF1, a component of the Cul4A E3 ubiquitin ligase. The capacity for Vpr to target PU.1 is highly conserved across primate lentiviruses. In addition to impacting infected cells, we find that Vpr suppresses expression of innate immune response genes in uninfected bystander cells, and that virion-associated Vpr can degrade PU.1. Together, we demonstrate Vpr counteracts PU.1 in macrophages to blunt antiviral immune responses and promote viral spread.
]]></description>
<dc:creator>Virgilio, M. C.</dc:creator>
<dc:creator>Disbennett, W. M.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Lubow, J.</dc:creator>
<dc:creator>Welch, J. D.</dc:creator>
<dc:creator>Collins, K. L.</dc:creator>
<dc:date>2023-03-21</dc:date>
<dc:identifier>doi:10.1101/2023.03.21.533528</dc:identifier>
<dc:title><![CDATA[HIV-1 Vpr combats the PU.1-driven antiviral response in primary human macrophages.]]></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/10.1101/2023.03.22.533820v1?rss=1">
<title>
<![CDATA[
Assessing vaccine-mediated protection in an ultra-low dose Mycobacterium tuberculosis murine model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.22.533820v1?rss=1</link>
<description><![CDATA[
Despite widespread immunization with Bacille-Calmette-Guerin (BCG), the only currently licensed tuberculosis (TB) vaccine, TB remains a leading cause of mortality globally. There are many TB vaccine candidates in the developmental pipeline, but the lack of a robust animal model to assess vaccine efficacy has hindered our ability to prioritize candidates for human clinical trials. Here we use a murine ultra-low dose (ULD) Mycobacterium tuberculosis (Mtb) challenge model to assess protection conferred by BCG vaccination. We show that BCG confers a reduction in lung bacterial burdens that is more durable than that observed after conventional dose challenge, curbs Mtb dissemination to the contralateral lung, and, in a small percentage of mice, prevents detectable infection. These findings are consistent with the ability of human BCG vaccination to mediate protection, particularly against disseminated disease, in specific human populations and clinical settings. Overall, our findings demonstrate that the ultra-low dose Mtb infection model can measure distinct parameters of immune protection that cannot be assessed in conventional dose murine infection models and could provide an improved platform for TB vaccine testing.
]]></description>
<dc:creator>Plumlee, C. R.</dc:creator>
<dc:creator>Barrett, H. W.</dc:creator>
<dc:creator>Shao, D. E.</dc:creator>
<dc:creator>Lien, K. A.</dc:creator>
<dc:creator>Cross, L. M.</dc:creator>
<dc:creator>Cohen, S. B.</dc:creator>
<dc:creator>Edlefsen, P. T.</dc:creator>
<dc:creator>Urdahl, K. B.</dc:creator>
<dc:date>2023-03-22</dc:date>
<dc:identifier>doi:10.1101/2023.03.22.533820</dc:identifier>
<dc:title><![CDATA[Assessing vaccine-mediated protection in an ultra-low dose Mycobacterium tuberculosis murine model]]></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/10.1101/2023.03.23.533974v1?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/10.1101/2023.03.24.534139v1?rss=1">
<title>
<![CDATA[
Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.24.534139v1?rss=1</link>
<description><![CDATA[
Human immunodeficiency virus (HIV) and other lentiviruses adapt to new hosts by evolving to evade host-specific innate immune proteins that differ in sequence and often viral recognition between host species. Understanding how these host antiviral proteins, called restriction factors, constrain lentivirus replication and transmission is key to understanding the emergence of pandemic viruses like HIV-1. Human TRIM34, a paralogue of the well-characterized lentiviral restriction factor TRIM5, was previously identified by our lab via CRISPR-Cas9 screening as a restriction factor of certain HIV and SIV capsids. Here, we show that diverse primate TRIM34 orthologues from non-human primates can restrict a range of Simian Immunodeficiency Virus (SIV) capsids including SIVAGM-SAB, SIVAGM-TAN and SIVMAC capsids, which infect sabaeus monkeys, tantalus monkeys, and rhesus macaques, respectively. All primate TRIM34 orthologues tested, regardless of species of origin, were able to restrict this same subset of viral capsids. However, in all cases, this restriction also required the presence of TRIM5. We demonstrate that TRIM5 is necessary, but not sufficient, for restriction of these capsids, and that human TRIM5 functionally interacts with TRIM34 from different species. Finally, we find that both the TRIM5 SPRY v1 loop and the TRIM34 SPRY domain are essential for TRIM34-mediated restriction. These data support a model in which TRIM34 is a broadly-conserved primate lentiviral restriction factor that acts in tandem with TRIM5, such that together, these proteins can restrict capsids that neither can restrict alone.
]]></description>
<dc:creator>Twentyman, J.</dc:creator>
<dc:creator>Khalifeh, A.</dc:creator>
<dc:creator>Felton, A.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Ohainle, M.</dc:creator>
<dc:date>2023-03-25</dc:date>
<dc:identifier>doi:10.1101/2023.03.24.534139</dc:identifier>
<dc:title><![CDATA[Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor]]></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/10.1101/2023.03.27.534417v1?rss=1">
<title>
<![CDATA[
Transcriptional Activation of Regenerative Hematopoiesis via Vascular Niche Sensing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.27.534417v1?rss=1</link>
<description><![CDATA[
Transition between activation and quiescence programs in hematopoietic stem and progenitor cells (HSC/HSPCs) is perceived to be governed intrinsically and by microenvironmental co-adaptation. However, HSC programs dictating both transition and adaptability, remain poorly defined. Single cell multiome analysis divulging differential transcriptional activity between distinct HSPC states, indicated for the exclusive absence of Fli-1 motif from quiescent HSCs. We reveal that Fli-1 activity is essential for HSCs during regenerative hematopoiesis. Fli-1 directs activation programs while manipulating cellular sensory and output machineries, enabling HSPCs co-adoptability with a stimulated vascular niche. During regenerative conditions, Fli-1 presets and enables propagation of niche-derived Notch1 signaling. Constitutively induced Notch1 signaling is sufficient to recuperate functional HSC impairments in the absence of Fli-1. Applying FLI-1 modified-mRNA transduction into lethargic adult human mobilized HSPCs, enables their vigorous niche-mediated expansion along with superior engraftment capacities. Thus, decryption of stem cell activation programs offers valuable insights for immune regenerative medicine.
]]></description>
<dc:creator>Itkin, T.</dc:creator>
<dc:creator>Houghton, S.</dc:creator>
<dc:creator>Schreiner, R.</dc:creator>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Badwe, C.</dc:creator>
<dc:creator>Voisin, V.</dc:creator>
<dc:creator>Murison, A.</dc:creator>
<dc:creator>Seyedhassantehrani, N.</dc:creator>
<dc:creator>Kaufmann, K. B.</dc:creator>
<dc:creator>Garcia-Prat, L.</dc:creator>
<dc:creator>Booth, G. T.</dc:creator>
<dc:creator>Geng, F.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Gomez-Salinero, J. M.</dc:creator>
<dc:creator>Shieh, J.-H.</dc:creator>
<dc:creator>Redmond, D.</dc:creator>
<dc:creator>Xiang, J. Z.</dc:creator>
<dc:creator>Josefowicz, S. Z.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Spencer, J. A.</dc:creator>
<dc:creator>Zangi, L.</dc:creator>
<dc:creator>Hadland, B.</dc:creator>
<dc:creator>Dick, J. E.</dc:creator>
<dc:creator>Xie, S. Z.</dc:creator>
<dc:creator>Rafii, S.</dc:creator>
<dc:date>2023-03-29</dc:date>
<dc:identifier>doi:10.1101/2023.03.27.534417</dc:identifier>
<dc:title><![CDATA[Transcriptional Activation of Regenerative Hematopoiesis via Vascular Niche Sensing]]></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/10.1101/2023.03.28.534648v1?rss=1">
<title>
<![CDATA[
Single-cell analysis of bidirectional reprogramming between early embryonic states reveals mechanisms of differential lineage plasticities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.28.534648v1?rss=1</link>
<description><![CDATA[
Two distinct fates, pluripotent epiblast (EPI) and primitive (extra-embryonic) endoderm (PrE), arise from common progenitor cells, the inner cell mass (ICM), in mammalian embryos. To study how these sister identities are forged, we leveraged embryonic (ES) and eXtraembryonic ENdoderm (XEN) stem cells - in vitro counterparts of the EPI and PrE. Bidirectional reprogramming between ES and XEN coupled with single-cell RNA and ATAC-seq analyses uncovered distinct rates, efficiencies and trajectories of state conversions, identifying drivers and roadblocks of reciprocal conversions. While GATA4-mediated ES-to-iXEN conversion was rapid and nearly deterministic, OCT4, KLF4 and SOX2-induced XEN-to-iPS reprogramming progressed with diminished efficiency and kinetics. The dominant PrE transcriptional program, safeguarded by Gata4, and globally elevated chromatin accessibility of EPI underscored the differential plasticities of the two states. Mapping in vitro trajectories to embryos revealed reprogramming in either direction tracked along, and toggled between, EPI and PrE in vivo states without transitioning through the ICM.
]]></description>
<dc:creator>Garg, V.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Nowotschin, S.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:creator>Kuo, Y.-Y.</dc:creator>
<dc:creator>Sharma, R.</dc:creator>
<dc:creator>Polyzos, A.</dc:creator>
<dc:creator>Salataj, E.</dc:creator>
<dc:creator>Murphy, D.</dc:creator>
<dc:creator>Jang, A.</dc:creator>
<dc:creator>Pe'er, D.</dc:creator>
<dc:creator>Apostolou, E.</dc:creator>
<dc:creator>Hadjantonakis, A.-K.</dc:creator>
<dc:date>2023-03-29</dc:date>
<dc:identifier>doi:10.1101/2023.03.28.534648</dc:identifier>
<dc:title><![CDATA[Single-cell analysis of bidirectional reprogramming between early embryonic states reveals mechanisms of differential lineage plasticities]]></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/10.1101/2023.04.07.535760v1?rss=1">
<title>
<![CDATA[
Stepwise design of pseudosymmetric protein hetero-oligomers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.07.535760v1?rss=1</link>
<description><![CDATA[
Pseudosymmetric hetero-oligomers with three or more unique subunits with overall structural (but not sequence) symmetry play key roles in biology, and systematic approaches for generating such proteins de novo would provide new routes to controlling cell signaling and designing complex protein materials. However, the de novo design of protein hetero-oligomers with three or more distinct chains with nearly identical structures is a challenging problem because it requires the accurate design of multiple protein-protein interfaces simultaneously. Here, we describe a divide-and-conquer approach that breaks the multiple-interface design challenge into a set of more tractable symmetric single-interface redesign problems, followed by structural recombination of the validated homo-oligomers into pseudosymmetric hetero-oligomers. Starting from de novo designed circular homo-oligomers composed of 9 or 24 tandemly repeated units, we redesigned the inter-subunit interfaces to generate 15 new homo-oligomers and recombined them to make 17 new hetero-oligomers, including ABC heterotrimers, A2B2 heterotetramers, and A3B3 and A2B2C2 heterohexamers which assemble with high structural specificity. The symmetric homo-oligomers and pseudosymmetric hetero-oligomers generated for each system share a common backbone, and hence are ideal building blocks for generating and functionalizing larger symmetric assemblies.

Significance StatementProtein oligomers composed of multiple unique subunits are versatile building blocks for creating functional materials and controlling biological processes. However, designing robust hetero-oligomers with distinct subunits and precise structural symmetry remains a major challenge. Here, we present a general strategy for designing such complexes by breaking down the problem into simpler steps by first symmetrically re-designing the interfaces of homo-oligomeric proteins, and then recombining validated variants to form pseudosymmetric hetero-oligomers. Using this method, we generated 17 hetero-oligomers with up to three unique subunits that assemble with high specificity. Our approach can be extended to create a wide range of pseudosymmetric assemblies for manipulating cellular signaling and as building blocks for advanced protein materials. These pseudosymmeteric heterotrimers have already enabled the construction of a set of massive nanocages, including a T=4 icosahedral nanocage with a 70 nm diameter and 240 subunits.1
]]></description>
<dc:creator>Kibler, R. D.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Kennedy, M. A.</dc:creator>
<dc:creator>Wicky, B. I. M.</dc:creator>
<dc:creator>Lai, S. M.</dc:creator>
<dc:creator>Kostelic, M. M.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Chow, C. M.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Wysocki, V. H.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2023-04-07</dc:date>
<dc:identifier>doi:10.1101/2023.04.07.535760</dc:identifier>
<dc:title><![CDATA[Stepwise design of pseudosymmetric protein hetero-oligomers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.07.536032v1?rss=1">
<title>
<![CDATA[
Rapid-kinetics degron benchmarking reveals off-target activities and mixed agonism-antagonism of MYB inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.07.536032v1?rss=1</link>
<description><![CDATA[
Attenuating aberrant transcriptional circuits holds great promise for the treatment of numerous diseases, including cancer. However, development of transcriptional inhibitors is hampered by the lack of a generally accepted functional cellular readout to characterize their target specificity and on-target activity. We benchmarked the direct gene-regulatory signatures of six agents reported as inhibitors of the oncogenic transcription factor MYB against targeted MYB degradation in a nascent transcriptomics assay. The inhibitors demonstrated partial specificity for MYB target genes but displayed significant off-target activity. Unexpectedly, the inhibitors displayed bimodal on-target effects, acting as mixed agonists-antagonists. Our data uncover unforeseen agonist effects of small molecules originally developed as TF inhibitors and argue that rapid-kinetics benchmarking against degron models should be used for functional characterization of transcriptional modulators.
]]></description>
<dc:creator>Harada, T.</dc:creator>
<dc:creator>Perez, M. W.</dc:creator>
<dc:creator>Kalfon, J.</dc:creator>
<dc:creator>Dievenich Braes, F.</dc:creator>
<dc:creator>Batley, R.</dc:creator>
<dc:creator>Eagle, K.</dc:creator>
<dc:creator>Nabet, B.</dc:creator>
<dc:creator>Leifer, B.</dc:creator>
<dc:creator>Kruell, J.</dc:creator>
<dc:creator>Paralkar, V. R.</dc:creator>
<dc:creator>Stegmaier, K.</dc:creator>
<dc:creator>Koehler, A. N.</dc:creator>
<dc:creator>Orkin, S. H.</dc:creator>
<dc:creator>Pimkin, M.</dc:creator>
<dc:date>2023-04-07</dc:date>
<dc:identifier>doi:10.1101/2023.04.07.536032</dc:identifier>
<dc:title><![CDATA[Rapid-kinetics degron benchmarking reveals off-target activities and mixed agonism-antagonism of MYB inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.11.536402v1?rss=1">
<title>
<![CDATA[
Identification of 1600 replication origins in S. cerevisiae 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.11.536402v1?rss=1</link>
<description><![CDATA[
There are approximately 500 known origins of replication in the yeast genome, and the process by which DNA replication initiates at these locations is well understood. In particular, these sites are made competent to initiate replication by loading of the Mcm replicative helicase prior to the start of S phase; thus, "a site to which MCM is bound in G1" might be considered to provide an operational definition of a replication origin. By fusing a subunit of Mcm to micrococcal nuclease, a technique referred to as "Chromatin Endogenous Cleavage", we previously showed that known origins are typically bound by a single Mcm double hexamer, loaded adjacent to the ARS consensus sequence (ACS). Here we extend this analysis from known origins to the entire genome, identifying candidate Mcm binding sites whose signal intensity varies over at least 3 orders of magnitude. Published data quantifying the production of ssDNA during S phase showed clear evidence of replication initiation among the most abundant 1600 of these sites, with replication activity decreasing in concert with Mcm abundance and disappearing at the limit of detection of ssDNA. Three other hallmarks of replication origins were apparent among the most abundant 5,500 sites. Specifically, these sites (1) appeared in intergenic nucleosome-free regions that were flanked on one or both sides by well-positioned nucleosomes; (2) were flanked by ACSs; and (3) exhibited a pattern of GC skew characteristic of replication initiation. Furthermore, the high resolution of this technique allowed us to demonstrate a strong bias for detecting Mcm double-hexamers downstream rather than upstream of the ACS, which is consistent with the directionality of Mcm loading by Orc that has been observed in vitro. We conclude that, if sites at which Mcm double-hexamers are loaded can function as replication origins, then DNA replication origins are at least 3-fold more abundant than previously assumed, and we suggest that replication may occasionally initiate in essentially every intergenic region. These results shed light on recent reports that as many as 15% of replication events initiate outside of known origins, and this broader distribution of replication origins suggest that S phase in yeast may be less distinct from that in humans than is widely assumed.
]]></description>
<dc:creator>Foss, E. J.</dc:creator>
<dc:creator>Lichauco, C.</dc:creator>
<dc:creator>Gatbonton-Schwager, T.</dc:creator>
<dc:creator>Lofts, B.</dc:creator>
<dc:creator>Lao, U.</dc:creator>
<dc:creator>Bedalov, A.</dc:creator>
<dc:date>2023-04-12</dc:date>
<dc:identifier>doi:10.1101/2023.04.11.536402</dc:identifier>
<dc:title><![CDATA[Identification of 1600 replication origins in S. cerevisiae]]></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/10.1101/2023.04.25.538357v1?rss=1">
<title>
<![CDATA[
Imetelstat-Mediated Alterations in Fatty Acid Metabolism To Induce Ferroptosis As Therapeutic Strategy for Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.25.538357v1?rss=1</link>
<description><![CDATA[
Telomerase enables replicative immortality in most cancers including acute myeloid leukemia (AML). Imetelstat is a first-in-class telomerase inhibitor with clinical efficacy in myelofibrosis and myelodysplastic syndromes. Here, we develop an AML patient-derived xenograft (PDX) resource, and perform integrated genomics, transcriptomics, and lipidomics analyses combined with functional genetics to identify key mediators of imetelstat efficacy. In a randomized Phase II-like preclinical trial in PDX, imetelstat effectively diminishes AML burden, and preferentially targets subgroups containing mutant NRAS and oxidative stress-associated gene expression signatures. Unbiased, genome-wide CRISPR/Cas9 editing identifies ferroptosis regulators as key mediators of imetelstat efficacy. Imetelstat promotes the formation of polyunsaturated fatty acid-containing phospholipids, causing excessive levels of lipid peroxidation and oxidative stress. Pharmacological inhibition of ferroptosis diminishes imetelstat efficacy. We leverage these mechanistic insights to develop an optimized therapeutic strategy using oxidative stress-inducing chemotherapy to sensitize patient samples to imetelstat causing significant disease control in AML.
]]></description>
<dc:creator>Bruedigam, C.</dc:creator>
<dc:creator>Porter, A. H.</dc:creator>
<dc:creator>Song, A.</dc:creator>
<dc:creator>Vroeg in de Wei, G.</dc:creator>
<dc:creator>Stoll, T.</dc:creator>
<dc:creator>Straube, J.</dc:creator>
<dc:creator>Cooper, L.</dc:creator>
<dc:creator>Cheng, G.</dc:creator>
<dc:creator>Kahl, V. F. S.</dc:creator>
<dc:creator>Sobinoff, A. P.</dc:creator>
<dc:creator>Ling, V. Y.</dc:creator>
<dc:creator>Jebaraj, B. M. C.</dc:creator>
<dc:creator>Janardhanan, Y.</dc:creator>
<dc:creator>Haldar, R.</dc:creator>
<dc:creator>Bray, L. J.</dc:creator>
<dc:creator>Bullinger, L.</dc:creator>
<dc:creator>Heidel, F. H.</dc:creator>
<dc:creator>Kennedy, G. A.</dc:creator>
<dc:creator>Hill, M. M.</dc:creator>
<dc:creator>Pickett, H. A.</dc:creator>
<dc:creator>Abdel-Wahab, O.</dc:creator>
<dc:creator>Hartel, G.</dc:creator>
<dc:creator>Lane, S. W.</dc:creator>
<dc:date>2023-04-28</dc:date>
<dc:identifier>doi:10.1101/2023.04.25.538357</dc:identifier>
<dc:title><![CDATA[Imetelstat-Mediated Alterations in Fatty Acid Metabolism To Induce Ferroptosis As Therapeutic Strategy for Acute Myeloid Leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.03.539298v1?rss=1">
<title>
<![CDATA[
Common Analysis of Direct RNA SequencinG CUrrently Leads to Misidentification of 5-Methylcytosine Modifications at GCU Motifs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.03.539298v1?rss=1</link>
<description><![CDATA[
RNA modifications, such as methylation, can be detected with Oxford Nanopore Technologies direct RNA sequencing. One commonly used tool for detecting 5-methylcytosine (m5C) modifications is Tombo, which uses an "Alternative Model" to detect putative modifications from a single sample. We examined direct RNA sequencing data from diverse taxa including virus, bacteria, fungi, and animals. The algorithm consistently identified a 5-methylcytosine at the central position of a GCU motif. However, it also identified a 5-methylcytosine in the same motif in fully unmodified in vitro transcribed RNA, suggesting that this a frequent false prediction. In the absence of further validation, several published predictions of 5-methylcytosine in human coronavirus and human cerebral organoid RNA in a GCU context should be reconsidered.

IMPORTANCEThe detection of chemical modifications to RNA is a rapidly expanding field within epigenetics. Nanopore sequencing technology provides an attractive means of detecting these modifications directly on the RNA, but accurate modification predictions are dependent upon the software developed to interpret the sequencing results. One of these tools, Tombo, allows users to detect modifications using sequencing results from a single RNA sample. However, we find that this method falsely predicts modifications in a specific sequence context across a variety of RNA samples, including RNA that lacks modifications. Results from previous publications include predictions in human coronaviruses with this sequence context and should be reconsidered. Our results highlight the importance of using RNA modification detection tools with caution in the absence of a control RNA sample for comparison.
]]></description>
<dc:creator>Watson, K.</dc:creator>
<dc:creator>Bromley, R.</dc:creator>
<dc:creator>Sparklin, B. C.</dc:creator>
<dc:creator>Gasser, M.</dc:creator>
<dc:creator>Bhattacharya, T.</dc:creator>
<dc:creator>Lebov, J. F.</dc:creator>
<dc:creator>Tyson, T.</dc:creator>
<dc:creator>Teigen, L.</dc:creator>
<dc:creator>Graf, K. T.</dc:creator>
<dc:creator>Michalski, M.</dc:creator>
<dc:creator>Bruno, V. M.</dc:creator>
<dc:creator>Lindsey, A. R. I.</dc:creator>
<dc:creator>Hardy, R. W.</dc:creator>
<dc:creator>Newton, I. L. G.</dc:creator>
<dc:creator>Dunning Hotopp, J. C.</dc:creator>
<dc:date>2023-05-03</dc:date>
<dc:identifier>doi:10.1101/2023.05.03.539298</dc:identifier>
<dc:title><![CDATA[Common Analysis of Direct RNA SequencinG CUrrently Leads to Misidentification of 5-Methylcytosine Modifications at GCU Motifs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.04.539469v1?rss=1">
<title>
<![CDATA[
CTCF-mediated insulation and chromatin environment modulate Car5b escape from X inactivation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.04.539469v1?rss=1</link>
<description><![CDATA[
BackgroundThe number and escape levels of genes that escape X chromosome inactivation (XCI) in female somatic cells vary among tissues and cell types, potentially contributing to specific sex differences. Here we investigate the role of CTCF, a master chromatin conformation regulator, in regulating escape from XCI. CTCF binding profiles and epigenetic features were systematically examined at constitutive and facultative escape genes using mouse allelic systems to distinguish the inactive X (Xi) and active X (Xa) chromosomes.

ResultsWe found that escape genes are located inside domains flanked by convergent arrays of CTCF binding sites, consistent with the formation of loops. In addition, strong and divergent CTCF binding sites often located at the boundaries between escape genes and adjacent neighbors subject to XCI would help insulate domains. Facultative escapees show clear differences in CTCF binding dependent on their XCI status in specific cell types/tissues. Concordantly, deletion but not inversion of a CTCF binding site at the boundary between the facultative escape gene Car5b and its silent neighbor Siah1b resulted in loss of Car5b escape. Reduced CTCF binding and enrichment of a repressive mark over Car5b in cells with a boundary deletion indicated loss of looping and insulation. In mutant lines in which either the Xi-specific compact structure or its H3K27me3 enrichment was disrupted, escape genes showed an increase in gene expression and associated active marks, supporting the roles of the 3D Xi structure and heterochromatic marks in constraining levels of escape.

ConclusionOur findings indicate that escape from XCI is modulated both by looping and insulation of chromatin via convergent arrays of CTCF binding sites and by compaction and epigenetic features of the surrounding heterochromatin.
]]></description>
<dc:creator>Fang, H.</dc:creator>
<dc:creator>Tronco, A. R.</dc:creator>
<dc:creator>Bonora, G.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Thakur, J.</dc:creator>
<dc:creator>Berletch, J. B.</dc:creator>
<dc:creator>Filippova, G. N.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:date>2023-05-04</dc:date>
<dc:identifier>doi:10.1101/2023.05.04.539469</dc:identifier>
<dc:title><![CDATA[CTCF-mediated insulation and chromatin environment modulate Car5b escape from X inactivation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.19.541367v1?rss=1">
<title>
<![CDATA[
An Atlas of Adaptive Evolution in Endemic Human Viruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.19.541367v1?rss=1</link>
<description><![CDATA[
Through antigenic evolution, viruses like seasonal influenza evade recognition by neutralizing antibodies elicited by previous infection or vaccination. This means that a person with antibodies well-tuned to an initial infection will not be protected against the same virus years later and that vaccine-mediated protection will decay. It is not fully understood which of the many endemic human viruses evolve in this fashion. To expand that knowledge, we assess adaptive evolution across the viral genome in 28 endemic viruses, spanning a wide range of viral families and transmission modes. We find that surface proteins consistently show the highest rates of adaptation, and estimate that ten viruses in this panel undergo antigenic evolution to selectively fix mutations that enable the virus to escape recognition by prior immunity. We compare overall rates of amino acid substitution between these antigenically-evolving viruses and SARS-CoV-2, showing that SARS-CoV-2 viruses are accumulating protein-coding changes at substantially faster rates than these endemic viruses.
]]></description>
<dc:creator>Kistler, K.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.19.541367</dc:identifier>
<dc:title><![CDATA[An Atlas of Adaptive Evolution in Endemic Human Viruses]]></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/10.1101/210468v1?rss=1">
<title>
<![CDATA[
Quantifying the ease of viral escape from broad and narrow antibodies to influenza hemagglutinin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/210468v1?rss=1</link>
<description><![CDATA[
Influenza virus can completely escape most antibodies with single mutations. However, rare antibodies broadly neutralize many viral strains. It is unclear how easily influenza virus might escape such antibodies if it was under strong pressure to do so. Here we map all single amino-acid mutations that increase resistance to broad antibodies targeting an H1 hemagglutinin. Crucially, our approach not only identifies antigenic mutations but also quantifies their effect sizes. All antibodies select mutations, but the effect sizes vary widely. The virus can escape a broad antibody that targets residues in hemagglutinins receptor-binding site the same way it escapes narrow strain-specific antibodies: via single mutations with huge effects. In contrast, broad antibodies targeting hemagglutinins stalk only select mutations with small effects. Therefore, among the antibodies we have examined, breadth is an imperfect indicator of the potential for viral escape via single mutations. Broadly neutralizing antibodies targeting the H1 hemagglutinin stalk are quantifiably harder to escape than the other antibodies tested here.
]]></description>
<dc:creator>Doud, M. B.</dc:creator>
<dc:creator>Lee, J. M.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2017-10-28</dc:date>
<dc:identifier>doi:10.1101/210468</dc:identifier>
<dc:title><![CDATA[Quantifying the ease of viral escape from broad and narrow antibodies to influenza hemagglutinin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/220285v1?rss=1">
<title>
<![CDATA[
Per-sample immunoglobulin germline inference from B cell receptor deep sequencing data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/220285v1?rss=1</link>
<description><![CDATA[
The collection of immunoglobulin genes in an individuals germline, which gives rise to B cell receptors via recombination, is known to vary significantly across individuals. In humans, for example, each individual has only a fraction of the several hundred known V alleles. Furthermore, the currently-accepted set of known V alleles is both incomplete (particularly for non-European samples), and contains a significant number of spurious alleles. The resulting uncertainty as to which immunoglobulin alleles are present in any given sample results in inaccurate B cell receptor sequence annotations, and in particular inaccurate inferred naive ancestors. In this paper we first show that the currently widespread practice of aligning each sequence to its closest match in the full set of IMGT alleles results in a very large number of spurious alleles that are not in the samples true set of germline V alleles. We then describe a new method for inferring each individuals germline gene set from deep sequencing data, and show that it improves upon existing methods by making a detailed comparison on a variety of simulated and real data samples. This new method has been integrated into the partis annotation and clonal family inference package, available at https://github.com/psathyrella/partis, and is run by default without affecting overall run time.nnAuthor SummaryAntibodies are an important component of the adaptive immune system, which itself determines our response to both pathogens and vaccines. They are produced by B cells through somatic recombination of germline DNA, which results in a vast diversity of antigen binding affinities across the B cell repertoire. We typically learn about the development of this repertoire, and its history of interaction with antigens, by sequencing large numbers of the DNA sequences from which antibodies are derived. In order to understand such data, it is necessary to determine the combination of germline V, D, and J genes that was rearranged to form each such B cell receptor sequence. This is difficult, however, because the immunoglobulin locus exhibits an extraordinary level of diversity across individuals - encompassing both allelic variation and gene duplication, deletion, and conversion - and because the locuss large size and repetitive structure make germline sequencing very difficult. In this paper we describe a new computational method that avoids this difficulty by inferring each individuals set of immunoglobulin germline genes directly from expressed B cell receptor sequence data.
]]></description>
<dc:creator>Ralph, D. K.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2017-11-16</dc:date>
<dc:identifier>doi:10.1101/220285</dc:identifier>
<dc:title><![CDATA[Per-sample immunoglobulin germline inference from B cell receptor deep sequencing data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/221747v1?rss=1">
<title>
<![CDATA[
Transcription activator-coactivator specificity is mediated by a large and dynamic fuzzy protein-protein complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/221747v1?rss=1</link>
<description><![CDATA[
Transcription activation domains (ADs) are inherently disordered proteins that often target multiple coactivator complexes, but the specificity of these interactions is not understood. Efficient activation by yeast Gcn4 requires tandem Gcn4 ADs and four activator-binding domains (ABDs) on its target, the Mediator subunit Med15. Multiple ABDs are a common feature of coactivator complexes. We find that the large Gcn4-Med15 complex is heterogeneous, containing nearly all possible AD-ABD interactions. This complex forms using a dynamic fuzzy protein-protein interface where ADs use hydrophobic residues to bind hydrophobic surfaces of the ABDs in multiple orientations. This combinatorial mechanism allows individual interactions of low affinity and specificity to generate a biologically functional, specific, and higher affinity complex despite lacking a defined protein-protein interface. This binding strategy is likely representative of many activators that target multiple coactivators and allows great flexibility in combinations of activators that synergize to regulate genes with variable coactivator requirements.
]]></description>
<dc:creator>Tuttle, L. M.</dc:creator>
<dc:creator>Pacheco, D.</dc:creator>
<dc:creator>Warfield, L.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Ranish, J.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:creator>Klevit, R. E.</dc:creator>
<dc:date>2017-11-18</dc:date>
<dc:identifier>doi:10.1101/221747</dc:identifier>
<dc:title><![CDATA[Transcription activator-coactivator specificity is mediated by a large and dynamic fuzzy protein-protein complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/223594v1?rss=1">
<title>
<![CDATA[
Developing a low-cost milliliter-scale chemostat array for precise control of cellular growth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/223594v1?rss=1</link>
<description><![CDATA[
Multiplexed milliliter-scale chemostats are useful for measuring cell physiology under various degrees of nutrient limitation and for experimental evolution. In each chemostat, fresh medium containing a growth rate-limiting metabolite is pumped into the culturing chamber at a constant rate, while culture effluent exits at an equal rate. Although such devices have been developed by various labs, key parameters - the accuracy and precision of flow rate and the operational range - are not explicitly characterized. Here we report the development of multiplexed milliliter-scale chemostats where flow rates for eight chambers can be independently controlled to vary within a wide range, corresponding to population doubling times of 3~ 13 hours. Importantly, flow rates are precise and accurate without the use of expensive feedback systems. Among the eight chambers, the maximal coefficient of variation in flow rate is less than 3%, and average flow rates are only slightly below targets, i.e., 3-6% for 13-hour and 0.6-1.0% for 3-hour doubling times. This deficit is largely due to evaporation and should be correctable. We experimentally demonstrate that our device allows accurate and precise quantification of population phenotypes.
]]></description>
<dc:creator>Skelding, D. H.</dc:creator>
<dc:creator>Hart, S. F. M.</dc:creator>
<dc:creator>Vidyasagar, T.</dc:creator>
<dc:creator>Pozhitkov, A. E.</dc:creator>
<dc:creator>Shou, W.</dc:creator>
<dc:date>2017-11-22</dc:date>
<dc:identifier>doi:10.1101/223594</dc:identifier>
<dc:title><![CDATA[Developing a low-cost milliliter-scale chemostat array for precise control of cellular growth]]></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/10.1101/223677v1?rss=1">
<title>
<![CDATA[
Cooperating H3N2 influenza virus variants are not detectable in primary clinical samples 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/223677v1?rss=1</link>
<description><![CDATA[
The high mutation rates of RNA viruses lead to rapid genetic diversification, which can enable cooperative interactions between variants in a viral population. We previously described two distinct variants of H3N2 influenza virus that cooperate in cell culture. These variants differ by a single mutation, D151G, in the neuraminidase protein. The D151G mutation reaches a stable frequency of about 50% when virus is passaged in cell culture. However, it is unclear whether selection for the cooperative benefits of D151G is a cell-culture phenomenon, or whether the mutation is also sometimes present at appreciable frequency in virus populations sampled directly from infected humans. Prior work has not detected D151G in unpassaged clinical samples, but these studies have used methods like Sanger sequencing and pyrosequencing that are relatively insensitive to low-frequency variation. We identified nine samples of human H3N2 influenza collected between 2013 to 2015 in which Sanger sequencing had detected a high frequency of the D151G mutation following one to three passages in cell culture. We deep-sequenced the unpassaged clinical samples to identify low-frequency viral variants. The frequency of D151G did not exceed the frequency of library preparation and sequencing errors in any of the sequenced samples. We conclude that passage in cell culture is primarily responsible for the frequent observations of D151G in recent H3N2 influenza strains.nnIMPORTANCEViruses mutate rapidly, and recent studies of RNA viruses have shown that related viral variants can sometimes cooperate to improve each others growth. We previously described two variants of H3N2 influenza virus that cooperate in cell culture. The mutation responsible for cooperation is often observed when human samples of influenza virus are grown in the lab before sequencing, but it is unclear whether the mutation also exists in human infections or is exclusively the result of lab passage. We identified nine human isolates of influenza that had developed the cooperating mutation after being grown in the lab, and performed highly sensitive deep-sequencing of the unpassaged clinical samples to determine whether the mutation existed in the original human infections. We found no evidence of the cooperating mutation in the unpassaged samples, suggesting that the cooperation primarily arises in laboratory conditions.
]]></description>
<dc:creator>Xue, K. S.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>Perez-Osorio, A.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2017-11-23</dc:date>
<dc:identifier>doi:10.1101/223677</dc:identifier>
<dc:title><![CDATA[Cooperating H3N2 influenza virus variants are not detectable in primary clinical samples]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/224048v1?rss=1">
<title>
<![CDATA[
Nextstrain: real-time tracking of pathogen evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/224048v1?rss=1</link>
<description><![CDATA[
SummaryUnderstanding the spread and evolution of pathogens is important for effective public health measures and surveillance. Nextstrain consists of a database of viral genomes, a bioinformatics pipeline for phylodynamics analysis, and an interactive visualisation platform. Together these present a real-time view into the evolution and spread of a range of viral pathogens of high public health importance. The visualization integrates sequence data with other data types such as geographic information, serology, or host species. Nextstrain compiles our current understanding into a single accessible location, publicly available for use by health professionals, epidemiologists, virologists and the public alike.nnAvailability and implementationAll code (predominantly JavaScript and Python) is freely available from github.com/nextstrain and the web-application is available at nextstrain.org.
]]></description>
<dc:creator>Hadfield, J.</dc:creator>
<dc:creator>Megill, C.</dc:creator>
<dc:creator>Bell, S. M.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Potter, B.</dc:creator>
<dc:creator>Callender, C.</dc:creator>
<dc:creator>Sagulenko, P.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:date>2017-11-22</dc:date>
<dc:identifier>doi:10.1101/224048</dc:identifier>
<dc:title><![CDATA[Nextstrain: real-time tracking of pathogen evolution]]></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/10.1101/228536v1?rss=1">
<title>
<![CDATA[
Transcription activation domains of the yeast factors Met4 and Ino2: tandem activation domains with properties similar to the yeast Gcn4 activator 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/228536v1?rss=1</link>
<description><![CDATA[
Eukaryotic transcription activation domains (ADs) are intrinsically disordered polypeptides that typically interact with coactivator complexes, leading to stimulation of transcription initiation, elongation and chromatin modifications. Here we examine the properties of two strong and conserved yeast ADs: Met4 and Ino2. Both factors have tandem ADs that were identified by conserved sequence and functional studies. While AD function from both factors depends on hydrophobic residues, Ino2 further requires key conserved acidic and polar residues for optimal function. Binding studies show that the ADs bind multiple Med15 activator binding domains (ABDs) with a similar order of micromolar affinity, and similar but distinct thermodynamic properties. Protein crosslinking shows that no unique complex is formed upon Met4-Med15 binding. Rather, we observed heterogeneous AD-ABD contacts with nearly every possible AD-ABD combination. Many of these properties are similar to those observed with the yeast activator Gcn4, which forms a large heterogeneous, dynamic, and fuzzy complex with Med15. We suggest that this molecular behavior is common among eukaryotic activators.
]]></description>
<dc:creator>Pacheco, D.</dc:creator>
<dc:creator>warfield, l.</dc:creator>
<dc:creator>brajcich, m.</dc:creator>
<dc:creator>robbins, h.</dc:creator>
<dc:creator>luo, j.</dc:creator>
<dc:creator>ranish, j.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:date>2017-12-04</dc:date>
<dc:identifier>doi:10.1101/228536</dc:identifier>
<dc:title><![CDATA[Transcription activation domains of the yeast factors Met4 and Ino2: tandem activation domains with properties similar to the yeast Gcn4 activator]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/235630v1?rss=1">
<title>
<![CDATA[
Mapping mutational effects along the evolutionary landscape of HIV envelope 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/235630v1?rss=1</link>
<description><![CDATA[
The immediate evolutionary space accessible to HIV is largely determined by how single amino-acid mutations affect fitness. These mutational effects can shift as the virus evolves. However, the prevalence of such shifts in mutational effects remains unclear. Here we quantify the effects on viral growth of all amino-acid mutations to two HIV envelope (Env) proteins that differ at >100 residues. Most mutations similarly affect both Envs, but the amino-acid preferences of a minority of sites have clearly shifted. These shifted sites usually prefer a specific amino acid in one Env, but tolerate many amino acids in the other. Surprisingly, shifts are only slightly enriched at sites that have substituted between the Envs -- and many occur at residues that do not even contact substitutions. Therefore, long-range epistasis can unpredictably shift Env's mutational tolerance during HIV evolution, although the amino-acid preferences of most sites are conserved between moderately diverged viral strains.
]]></description>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Hilton, S. K.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2017-12-16</dc:date>
<dc:identifier>doi:10.1101/235630</dc:identifier>
<dc:title><![CDATA[Mapping mutational effects along the evolutionary landscape of HIV envelope]]></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/10.1101/255133v1?rss=1">
<title>
<![CDATA[
Viral diversity is an obligate consideration in CRISPR/Cas9 designs for HIV cure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/255133v1?rss=1</link>
<description><![CDATA[
RNA-guided CRISPR/Cas9 systems can be designed to mutate or excise the integrated HIV genome from latently infected cells and have therefore been proposed as a curative approach for HIV. However, most studies to date have focused on molecular clones with ideal target site recognition and do not account for target site variability observed within and between patients. For clinical success and broad applicability, guide RNA (gRNA) selection must account for circulating strain diversity and incorporate the within-host diversity of HIV. To address this, we identified a set of gRNAs targeting HIV LTR, gag and pol using publicly available sequences for these genes. We ranked gRNAs according to global conservation across HIV-1 group M and within subtypes A-C. By considering paired and triplet combinations of gRNAs, we found triplet sets of target sites such that at least one of the gRNAs in the set was present in over 98% of all globally-available sequences. We then selected 59 gRNAs from our list of highly-conserved LTR target sites and evaluated in vitro activity using a loss-of-function LTR-GFP fusion reporter. We achieved efficient GFP knockdown with multiple gRNAs and found clustering of highly active gRNA target sites near the middle of the LTR. Using published deep-sequence data from HIV-infected patients, we found that globally conserved sites also had greater within-host target conservation. Lastly, we developed a mathematical model based on varying distributions of within-host HIV sequence diversity and enzyme efficacy. We used the model to estimate the number of doses required to deplete the latent reservoir and achieve functional cure thresholds. Our modeling results highlight the importance of within-host target site conservation. While increased doses may overcome low target cleavage efficiency, inadequate targeting of rare strains is predicted to lead to rebound upon ART cessation even with many doses.nnAuthor summaryThe field of genome engineering has exploded over the last decade with the discovery of targeted endonucleases such as CRISPR/Cas9. Endonucleases are now being used to develop a wide range of therapeutics and their use has expanded into antiviral therapy against latent viral infections like HIV. The idea is to induce mutations in latent viral genomes that will render them replication-incompetent, thereby producing a functional cure. Although a great deal of progress has been made, most studies to date have relied on molecular clones that represent "ideal" targets. For clinical success and broad applicability, these therapies need to account for viral genetic diversity within and between individuals. Our paper examines the impact of HIV diversity on CRISPR-based cure strategies to determine the predictors of future clinical success. We performed an exhaustive and detailed computational analysis to identify optimal CRISPR target sites, taking into consideration both within-host and global viral diversity. We coupled this with laboratory testing of highly-conserved guides and compared measured activity to predicted results. Finally, we developed a mathematical model to predict the impact of enzyme activity and viral diversity on the number of doses of a CRISPR-based therapy needed to achieve a functional cure of HIV.
]]></description>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>De Silva Feelixge, H.</dc:creator>
<dc:creator>Reeves, D. B.</dc:creator>
<dc:creator>Mayer, B. T.</dc:creator>
<dc:creator>Stone, D.</dc:creator>
<dc:creator>Schiffer, J. T.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:date>2018-01-27</dc:date>
<dc:identifier>doi:10.1101/255133</dc:identifier>
<dc:title><![CDATA[Viral diversity is an obligate consideration in CRISPR/Cas9 designs for HIV cure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/262519v1?rss=1">
<title>
<![CDATA[
Cryopreservation of human mucosal tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/262519v1?rss=1</link>
<description><![CDATA[
BACKGROUNDCryopreservation of leukocytes isolated from the cervicovaginal and colorectal mucosa is useful for the study of cellular immunity (see Hughes SM et al. PLOS ONE 2016). However, some questions about mucosal biology and sexually transmitted infections are better addressed with intact mucosal tissue, for which there is no standard cryopreservation protocol.nnMETHODS AND FINDINGSTo find an optimal preservation protocol for mucosal tissues, we tested slow cooling (1%C/min) with 10% dimethylsulfoxide (designated "cryopreservation") and fast cooling (plunge in liquid nitrogen) with 20% dimethylsulfoxide and 20% ethylene glycol ("vitrification"). We compared fresh and preserved human cervicovaginal and colorectal tissues in a range of assays, including metabolic activity, human immunodeficiency virus infection, cell phenotype, tissue structure by hematoxylin-and-eosin staining, cell number and viability, production of cytokines, and microbicide drug concentrations. Metabolic activity, HIV infectability, and tissue structure were similar in cryopreserved and vitrified vaginal tissues. However, vitrification led to poor cell recovery from the colorectal mucosa, with 90% fewer cells recovered after isolation from vitrified colorectal tissues than from cryopreserved. HIV infection rates were similar for fresh and cryopreserved ectocervical tissues, whereas cryopreserved colorectal tissues were less easily infected than fresh tissues (hazard ratio 0.7 [95% confidence interval 0.4, 1.2]). Finally, we compared isolation of cells before and after cryopreservation. Cell recoveries were higher when cells were isolated after freezing and thawing (71% [59-84%]) than before (50% [38-62%]). Cellular function was similar to fresh tissue in both cases. Microbicide drug concentrations were lower in cryopreserved explants compared to fresh ones.nnCONCLUSIONSCryopreservation of intact cervicovaginal and colorectal tissues with dimethylsulfoxide works well in a range of assays, while the utility of vitrification is more limited. Cell yields are higher from cryopreserved intact tissue pieces than from thawed cryopreserved single cell suspensions isolated before freezing, but T cell functions are similar.
]]></description>
<dc:creator>Hughes, S. M.</dc:creator>
<dc:creator>Ferre, A. L.</dc:creator>
<dc:creator>Yandura, S. E.</dc:creator>
<dc:creator>Shetler, C.</dc:creator>
<dc:creator>Baker, C. A. R.</dc:creator>
<dc:creator>Calienes, F.</dc:creator>
<dc:creator>Levy, C. N.</dc:creator>
<dc:creator>Astronomo, R. D.</dc:creator>
<dc:creator>Shu, Z.</dc:creator>
<dc:creator>Lentz, G. M.</dc:creator>
<dc:creator>Fialkow, M.</dc:creator>
<dc:creator>Kirby, A. C.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Sinclair, E.</dc:creator>
<dc:creator>Rohan, L. C.</dc:creator>
<dc:creator>Anderson, P. L.</dc:creator>
<dc:creator>Shacklett, B. L.</dc:creator>
<dc:creator>Dezzutti, C. S.</dc:creator>
<dc:creator>Gao, D.</dc:creator>
<dc:creator>Hladik, F.</dc:creator>
<dc:date>2018-03-14</dc:date>
<dc:identifier>doi:10.1101/262519</dc:identifier>
<dc:title><![CDATA[Cryopreservation of human mucosal tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/271114v1?rss=1">
<title>
<![CDATA[
Seasonal influenza circulation patterns and projections for Feb 2018 to Feb 2019 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/271114v1?rss=1</link>
<description><![CDATA[
This report details current seasonal influenza circulation patterns as of Feb 2018 and makes projections up to Feb 2019 to coincide with selection of the 2018-2019 Northern Hemisphere vaccine strain. This is not meant as a comprehensive report, but is instead intended as particular observations that weve made that may be of relevance. Please also note that observed patterns reflect the GISAID database and may not be entirely representative of underlying dynamics. All analyses are based on the nextflu pipeline [1] with continual updates posted to nextflu.org.nnA/H3N2: H3N2 diversity has largely been replaced by subclades A1b, A2 and A3 within 3c2.A. Subclades A1b and A2 predominate in the population and each shows increases in frequency, mutations at epitope sites and evidence for minor changes to antigenic phenotype. Clade A1b may be marginally fitter than clade A2, but we expect both clades to persist into the future without a clear immediate winner.nnA/H1N1pdm: A clade comprising mutations S74R, S164T and I295V has recently swept to fixation. The rapidity of this sweep suggests a selective origin. However, there is no evidence of antigenic change.nnB/Vic: Very little B/Vic activity has been observed in recent months. A clade with a two codon deletion at sites HA1:162/163 has gradually risen in frequency. HI measurements suggest an 8 to 16-fold titer drop relative to the vaccine strain, but this antigenic change has not yet resulted in a rapid rise of this variant.nnB/Yam: Europe experienced a strong and early B/Yam season in absence of amino acid variation in HA or antigenic diversity. However, several mutations in NA have rapidly swept or risen to intermediate frequencies.
]]></description>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Neher, R.</dc:creator>
<dc:date>2018-02-25</dc:date>
<dc:identifier>doi:10.1101/271114</dc:identifier>
<dc:title><![CDATA[Seasonal influenza circulation patterns and projections for Feb 2018 to Feb 2019]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/291971v1?rss=1">
<title>
<![CDATA[
Isw2 and Ino80 chromatin remodeling factors regulate chromatin, replication, and copy number at the yeast ribosomal DNA locus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/291971v1?rss=1</link>
<description><![CDATA[
In the budding yeast Saccharomyces cerevisiae, ribosomal RNA genes are encoded in a highly repetitive tandem array referred to as the ribosomal DNA (rDNA) locus. The yeast rDNA is the site of a diverse set of DNA-dependent processes, including transcription of ribosomal RNAs by RNA Polymerases I and III, transcription of non-coding RNAs by RNA Polymerase II, DNA replication initiation, replication fork blocking, and recombination-mediated regulation of rDNA repeat copy number. All of this takes place in the context of chromatin, but relatively little is known about the roles played by ATP-dependent chromatin remodeling factors at the yeast rDNA. In this work, we report that the Isw2 and Ino80 chromatin remodeling factors are targeted to this highly repetitive locus. We characterize for the first time their function in modifying local chromatin structure, finding that loss of these factors affects the occupancy of nucleosomes in the 35S ribosomal RNA gene and the positioning of nucleosomes flanking the ribosomal origin of replication. In addition, we report that Isw2 and Ino80 promote efficient firing of the ribosomal origin of replication and facilitate the regulated increase of rDNA repeat copy number. This work significantly expands our understanding of the importance of ATP-dependent chromatin remodeling for rDNA biology.nnAuthor SummaryTo satisfy high cellular demand for ribosomes, genomes contain many copies of the genes encoding the RNA components of ribosomes. In the budding yeast Saccharomyces cerevisiae, these ribosomal RNA genes are located in the "ribosomal DNA locus", a highly repetitive array that contains approximately 150 copies of the same unit, in contrast to the single copies that suffice for most genes. This repetitive quality creates unique regulatory needs. Chromatin structure, the packaging and organization of DNA, is a critical determinant of DNA-dependent processes throughout the genome. ATP-dependent chromatin remodeling factors are important regulators of chromatin structure, and yet relatively little is known about how members of this class of protein affect DNA organization or behavior at the rDNA. In this work, we show that the Isw2 and Ino80 chromatin remodeling factors regulate two features of chromatin structure at the rDNA, the occupancy and the positioning of nucleosomes. In addition, we find that these factors regulate two critical processes that function uniquely at this locus: DNA replication originating from within the rDNA array, and the regulated increase of rDNA repeat copy number.
]]></description>
<dc:creator>Cutler, S.</dc:creator>
<dc:creator>Lee, L. J.</dc:creator>
<dc:creator>Tsukiyama, T.</dc:creator>
<dc:date>2018-03-30</dc:date>
<dc:identifier>doi:10.1101/291971</dc:identifier>
<dc:title><![CDATA[Isw2 and Ino80 chromatin remodeling factors regulate chromatin, replication, and copy number at the yeast ribosomal DNA locus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/293639v1?rss=1">
<title>
<![CDATA[
Estimating the frequency of multiplets in single-cell RNA sequencing from cell-mixing experiments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/293639v1?rss=1</link>
<description><![CDATA[
In single-cell RNA-sequencing, it is important to know the frequency at which the sequenced transcriptomes actually derive from multiple cells. A common method to estimate this multiplet frequency is to mix two different types of cells (e.g., human and mouse), and then determine how often the transcriptomes contain transcripts from both cell types. When the two cell types are mixed in equal proportion, the calculation of the multiplet frequency from the frequency of mixed transcriptomes is straightforward. But surprisingly, there are no published descriptions of how to calculate the multiplet frequency in the general case when the cell types are mixed unequally. Here I derive equations to analytically calculate the multiplet frequency from the numbers of observed pure and mixed transcriptomes when two cell types are mixed in arbitrary proportions, under the assumption that the loading of cells into droplets or wells is Poisson.
]]></description>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2018-04-02</dc:date>
<dc:identifier>doi:10.1101/293639</dc:identifier>
<dc:title><![CDATA[Estimating the frequency of multiplets in single-cell RNA sequencing from cell-mixing experiments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/295352v1?rss=1">
<title>
<![CDATA[
Functional Analysis of Metagenomes by Likelihood Inference (FAMLI) Successfully Compensates for Multi-Mapping Short Reads from Metagenomic Samples 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/295352v1?rss=1</link>
<description><![CDATA[
High-throughput sequencing can establish the functional capacity of a microbial community by cataloging the protein-coding sequences (CDS) present in the metagenome of the community. The relative performance of different computational methods for identifying CDS from whole-genome shotgun sequencing (WGS) is not fully established.nnHere we present an automated benchmarking workflow, using synthetic shotgun sequencing reads for which we know the true CDS content of the underlying communities, to determine the relative performance (sensitivity, positive predictive value or PPV, and computational efficiency) of different metagenome analysis tools for extracting the CDS content of a microbial community.nnAssembly-based methods are limited by coverage depth, with poor sensitivity for CDS at < 5X depth of sequencing, but have excellent PPV. Mapping-based techniques are more sensitive at low coverage depths, but can struggle with PPV. We additionally describe an expectation maximization based iterative algorithmic approach which we show to successfully improve the PPV of a mapping based technique while retaining improved sensitivity and computational efficiency.
]]></description>
<dc:creator>Golob, J. L.</dc:creator>
<dc:creator>Minot, S. S.</dc:creator>
<dc:date>2018-04-05</dc:date>
<dc:identifier>doi:10.1101/295352</dc:identifier>
<dc:title><![CDATA[Functional Analysis of Metagenomes by Likelihood Inference (FAMLI) Successfully Compensates for Multi-Mapping Short Reads from Metagenomic Samples]]></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/10.1101/298364v1?rss=1">
<title>
<![CDATA[
Deep mutational scanning of hemagglutinin helps predict evolutionary fates of human H3N2 influenza variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/298364v1?rss=1</link>
<description><![CDATA[
Human influenza virus rapidly accumulates mutations in its major surface protein hemagglutinin (HA). The evolutionary success of influenza virus lineages depends on how these mutations affect HAs functionality and antigenicity. Here we experimentally measure the effects on viral growth in cell culture of all single amino-acid mutations to the HA from a recent human H3N2 influenza virus strain. We show that mutations that are measured to be more favorable for viral growth are enriched in evolutionarily successful H3N2 viral lineages relative to mutations that are measured to be less favorable for viral growth. Therefore, despite the well-known caveats about cell-culture measurements of viral fitness, such measurements can still be informative for understanding evolution in nature. We also compare our measurements for H3 HA to similar data previously generated for a distantly related H1 HA, and find substantial differences in which amino acids are preferred at many sites. For instance, the H3 HA has less disparity in mutational tolerance between the head and stalk domains than the H1 HA. Overall, our work suggests that experimental measurements of mutational effects can be leveraged to help understand the evolutionary fates of viral lineages in nature -- but only when the measurements are made on a viral strain similar to the ones being studied in nature.nnSignificance StatementA key goal in the study of influenza virus evolution is to forecast which viral strains will persist and which ones will die out. Here we experimentally measure the effects of all amino-acid mutations to the hemagglutinin protein from a human H3N2 influenza strain on viral growth in cell culture. We show that these measurements have utility for distinguishing among viral strains that do and do not succeed in nature. Overall, our work suggests that new high-throughput experimental approaches may be useful for understanding virus evolution in nature.
]]></description>
<dc:creator>Lee, J. M.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Doud, M. B.</dc:creator>
<dc:creator>Hooper, K. A.</dc:creator>
<dc:creator>Wu, N. C.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2018-04-10</dc:date>
<dc:identifier>doi:10.1101/298364</dc:identifier>
<dc:title><![CDATA[Deep mutational scanning of hemagglutinin helps predict evolutionary fates of human H3N2 influenza variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/298737v1?rss=1">
<title>
<![CDATA[
EZH2 co-opts gain-of-function p53 mutants to promote cancer growth and metastasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/298737v1?rss=1</link>
<description><![CDATA[
With the unfolding of more and more cancer-driven gain-of-function (GOF) mutants of p53, it is important to define a common mechanism to systematically target different mutants rather than develop strategies tailored to inhibit each mutant individually. Here, using RNA immunoprecipitation sequencing (RIP-seq) we identified EZH2 as a p53 mRNA-binding protein. EZH2 bound to the internal ribosome entry site (IRES) in the 5 untranslated region (5UTR) of p53 mRNA and enhanced p53 protein translation in a methyltransferase-independent manner. EZH2 augmented p53 GOF mutant-mediated cancer growth and metastasis by increasing p53 GOF mutant protein level. EZH2 overexpression associated with the worse outcome only in patients with p53-mutated cancer. Depletion of EZH2 by antisense oligonucleotides inhibited p53 GOF mutant-mediated cancer growth. Our findings reveal a non-methyltransferase function of EZH2 that controls protein translation of p53 GOF mutants, inhibition of which causes synthetic lethality in cancer cells expressing p53 GOF mutants.
]]></description>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Ding, L.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Ye, Z.</dc:creator>
<dc:creator>Pan, Y.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Coleman, R.</dc:creator>
<dc:creator>Coleman, I.</dc:creator>
<dc:creator>Karnes, R. J.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Zhu, R.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:date>2018-04-10</dc:date>
<dc:identifier>doi:10.1101/298737</dc:identifier>
<dc:title><![CDATA[EZH2 co-opts gain-of-function p53 mutants to promote cancer growth and metastasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/302703v1?rss=1">
<title>
<![CDATA[
Modeling site-specific amino-acid preferences deepens phylogenetic estimates of viral divergence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/302703v1?rss=1</link>
<description><![CDATA[
Molecular phylogenetics is often used to estimate the time since the divergence of modern gene sequences. For highly diverged sequences, such phylogenetic techniques sometimes estimate surprisingly recent divergence times. In the case of viruses, independent evidence indicates that the estimates of deep divergence times from molecular phylogenetics are sometimes too recent. This discrepancy is caused in part by inadequate models of purifying selection leading to branch-length underestimation. Here we examine the effect on branch-length estimation of using models that incorporate experimental measurements of purifying selection. We find that models informed by experimentally measured site-specific amino-acid preferences estimate longer deep branches on phylogenies of influenza virus hemagglutinin. This lengthening of branches is due to more realistic stationary states of the models, and is mostly independent of the branch-length-extension from modeling site-to-site variation in amino-acid substitution rate. The branch-length extension from experimentally informed site-specific models is similar to that achieved by other approaches that allow the stationary state to vary across sites. However, the improvements from all of these site-specific but time-homogeneous and site-independent models are limited by the fact that a proteins amino-acid preferences gradually shift as it evolves. Overall, our work underscores the importance of modeling site-specific amino-acid preferences when estimating deep divergence times--but also shows the inherent limitations of approaches that fail to account for how these preferences shift over time.
]]></description>
<dc:creator>Hilton, S. K.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2018-04-17</dc:date>
<dc:identifier>doi:10.1101/302703</dc:identifier>
<dc:title><![CDATA[Modeling site-specific amino-acid preferences deepens phylogenetic estimates of viral divergence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/307587v1?rss=1">
<title>
<![CDATA[
Complete functional mapping of infection- and vaccine-elicited antibodies against the fusion peptide of HIV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/307587v1?rss=1</link>
<description><![CDATA[
Eliciting broadly neutralizing antibodies (bnAbs) targeting envelope (Env) is a major goal of HIV vaccine development, but cross-clade breadth from immunization has only sporadically been observed. Recently, Xu et al (2018) elicited cross-reactive neutralizing antibody responses in a variety of animal models using immunogens based on the epitope of bnAb VRC34.01. The VRC34.01 antibody, which was elicited by natural human infection, targets the N terminus of the Env fusion peptide, a critical component of the virus entry machinery. Here we precisely characterize the functional epitopes of VRC34.01 and two vaccine-elicited murine antibodies by mapping all single amino-acid mutations to the BG505 Env that affect viral neutralization. While escape from VRC34.01 occurred via mutations in both fusion peptide and distal interacting sites of the Env trimer, escape from the vaccine-elicited antibodies was mediated predominantly by mutations in the fusion peptide. Cryo-electron microscopy of four vaccine-elicited antibodies in complex with Env trimer revealed focused recognition of the fusion peptide and provided a structural basis for development of neutralization breadth. Together, these functional and structural data suggest that the breadth of vaccine-elicited antibodies targeting the fusion peptide can be enhanced by specific interactions with additional portions of Env. Thus, our complete maps of viral escape provide a template to improve the breadth or potency of future vaccine-induced antibodies against Envs fusion peptide.nnAuthor summaryA major goal of HIV-1 vaccine design is to elicit antibodies that neutralize diverse strains of HIV-1. Recently, some of us elicited such antibodies in animal models using immunogens based on the epitope of a broad antibody (VRC34.01) isolated from an infected individual. Further improving these vaccine-elicited antibody responses will require a detailed understanding of how the resulting antibodies target HIVs envelope protein (Env). Here, we used mutational antigenic profiling to precisely map the epitope of two vaccine-elicited antibodies and the template VRC34.01 antibody. We did this by quantifying the effect of all possible amino acid mutations to Env on antibody neutralization. Although all antibodies target a similar region of Env, we found clear differences in the functional interaction of Env with the vaccine- and infection-elicited antibodies. We combined these functional data with structural analyses to identify antibody-Env interactions that could improve the breadth of vaccine-elicited antibodies, and thereby help to refine vaccination schemes to achieve broader responses.
]]></description>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Acharya, P.</dc:creator>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Rawi, R.</dc:creator>
<dc:creator>Xu, K.</dc:creator>
<dc:creator>Chuang, G.-Y.</dc:creator>
<dc:creator>Wei, H.</dc:creator>
<dc:creator>Mascola, J. R.</dc:creator>
<dc:creator>Carragher, B.</dc:creator>
<dc:creator>Potter, C. S.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Kwong, P. D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2018-04-24</dc:date>
<dc:identifier>doi:10.1101/307587</dc:identifier>
<dc:title><![CDATA[Complete functional mapping of infection- and vaccine-elicited antibodies against the fusion peptide of HIV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/307736v1?rss=1">
<title>
<![CDATA[
Benchmarking tree and ancestral sequence inference for B cell receptor sequences 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/307736v1?rss=1</link>
<description><![CDATA[
B cell receptor sequences evolve during affinity maturation according to a Darwinian process of mutation and selection. Phylogenetic tools are used extensively to reconstruct ancestral sequences and phylogenetic trees from affinity-matured sequences. In addition to using general-purpose phylogenetic methods, researchers have developed new tools to accommodate the special features of B cell sequence evolution. However, the performance of classical phylogenetic techniques in the presence of B cell-specific features is not well understood, nor how much the newer generation of B cell specific tools represent an improvement over classical methods. In this paper we benchmark the performance of classical phylogenetic and new B cell-specific tools when applied to B cell receptor sequences simulated from a forward-time model of B cell receptor affinity maturation towards a mature receptor. We show that the currently used tools vary substantially in terms of tree structure and ancestral sequence inference accuracy. Furthermore, we show that there are still large performance gains to be achieved by modeling the special mutation process of B cell receptors. These conclusions are further strengthened with real data using the rules of isotype switching to count possible violations within each inferred phylogeny.
]]></description>
<dc:creator>Davidsen, K.</dc:creator>
<dc:creator>Matsen, F.</dc:creator>
<dc:date>2018-04-25</dc:date>
<dc:identifier>doi:10.1101/307736</dc:identifier>
<dc:title><![CDATA[Benchmarking tree and ancestral sequence inference for B cell receptor sequences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/313106v1?rss=1">
<title>
<![CDATA[
Human T cell receptor occurrence patterns encode immune history, genetic background, and receptor specificity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/313106v1?rss=1</link>
<description><![CDATA[
The T cell receptor (TCR) repertoire encodes immune exposure history through the dynamic formation of immunological memory. Statistical analysis of repertoire sequencing data has the potential to decode disease associations from large cohorts with measured phenotypes. However, the repertoire perturbation induced by a given immunological challenge is conditioned on genetic background via major histocompatibility complex (MHC) polymorphism. We explore associations between MHC alleles, immune exposures, and shared TCRs in a large human cohort. Using a previously published repertoire sequencing dataset augmented with high-resolution MHC genotyping, our analysis reveals rich structure: striking imprints of common pathogens, clusters of co-occurring TCRs that may represent markers of shared immune exposures, and substantial variations in TCR-MHC association strength across MHC loci. Guided by atomic contacts in solved TCR:peptide-MHC structures, we identify sequence covariation between TCR and MHC. These insights and our analysis framework lay the groundwork for further explorations into TCR diversity.
]]></description>
<dc:creator>DeWitt, W. S.</dc:creator>
<dc:creator>Smith, A.</dc:creator>
<dc:creator>Schoch, G.</dc:creator>
<dc:creator>Hansen, J. A.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Bradley, P. H.</dc:creator>
<dc:date>2018-05-02</dc:date>
<dc:identifier>doi:10.1101/313106</dc:identifier>
<dc:title><![CDATA[Human T cell receptor occurrence patterns encode immune history, genetic background, and receptor specificity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/320895v1?rss=1">
<title>
<![CDATA[
Condensin-dependent chromatin condensation represses transcription globally during quiescence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/320895v1?rss=1</link>
<description><![CDATA[
Quiescence is a stress-resistant state in which cells reversibly exit the mitotic cell cycle and suspend most cellular processes. Quiescence is essential for stem cell maintenance and its misregulation is implicated in tumor formation. One of the conserved hallmarks of quiescent cells, from Saccharomyces cerevisiae to humans, is highly condensed chromatin. Here, we use Micro-C XL to map chromatin contacts at single-nucleosome resolution genome-wide to elucidate mechanisms and functions of condensed chromatin in quiescent S. cerevisiae cells. We describe previously uncharacterized chromatin domains on the order of 10-60 kilobases that in quiescent cells are formed by condensin-mediated chromatin loops. Conditional depletion of condensin prevents chromatin condensation during quiescence entry and leads to widespread transcriptional de-repression. We further demonstrate that condensin-dependent chromatin compaction is conserved in quiescent human fibroblasts. We propose that condensin-dependent condensation of chromatin represses transcription throughout the quiescent cell genome.
]]></description>
<dc:creator>Swygert, S. G.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Fu, T.</dc:creator>
<dc:creator>Hsieh, T.-H.</dc:creator>
<dc:creator>Rando, O. J.</dc:creator>
<dc:creator>Eisenman, R. N.</dc:creator>
<dc:creator>Schendure, J.</dc:creator>
<dc:creator>McKnight, J. N.</dc:creator>
<dc:creator>Tsukiyama, T.</dc:creator>
<dc:date>2018-05-12</dc:date>
<dc:identifier>doi:10.1101/320895</dc:identifier>
<dc:title><![CDATA[Condensin-dependent chromatin condensation represses transcription globally during quiescence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/321448v1?rss=1">
<title>
<![CDATA[
Translational control through differential ribosome pausing during amino acid limitation in mammalian cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/321448v1?rss=1</link>
<description><![CDATA[
Limitation for amino acids is thought to regulate translation in mammalian cells primarily by signaling through the kinases mTORC1 and GCN2. We find that limitation for the amino acid arginine causes a selective loss of tRNA charging, which regulates translation through ribosome pausing at two of six arginine codons. Surprisingly, limitation for leucine, an essential and abundant amino acid in protein, results in little or no ribosome pausing. Chemical and genetic perturbation of mTORC1 and GCN2 signaling revealed that their robust response to leucine limitation prevents ribosome pausing, while an insufficient response to arginine limitation led to loss of arginine tRNA charging and ribosome pausing. Codon-specific ribosome pausing decreased protein production and triggered premature ribosome termination without significantly reducing mRNA levels. Together, our results suggest that amino acids which are not optimally sensed by the mTORC1 and GCN2 pathways still regulate translation through an evolutionarily conserved mechanism based on codon-specific ribosome pausing.
]]></description>
<dc:creator>Darnell, A. M.</dc:creator>
<dc:creator>Subramaniam, A. R.</dc:creator>
<dc:creator>O'Shea, E. K.</dc:creator>
<dc:date>2018-05-14</dc:date>
<dc:identifier>doi:10.1101/321448</dc:identifier>
<dc:title><![CDATA[Translational control through differential ribosome pausing during amino acid limitation in mammalian cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/322420v1?rss=1">
<title>
<![CDATA[
Connectivity-Informed Adaptive Regularization for Generalized Outcomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/322420v1?rss=1</link>
<description><![CDATA[
One of the challenging problems in the brain imaging research is a principled incorporation of information from different imaging modalities in association studies. Frequently, data from each modality is analyzed separately using, for instance, dimensionality reduction techniques, which result in a loss of mutual information. We propose a novel regularization method, griPEER (generalized ridgified Partially Empirical Eigenvectors for Regression) to estimate the association between the brain structure features and a scalar outcome within the generalized linear regression framework. griPEER provides a principled approach to use external information from the structural brain connectivity to improve the regression coefficient estimation. Our proposal incorporates a penalty term, derived from the structural connectivity Laplacian matrix, in the penalized generalized linear regression. We address both theoretical and computational issues and show that our method is robust to the incomplete information about the structural brain connectivity. We also provide a significance testing procedure for performing inference on the estimated coefficients in this model. griPEER is evaluated in extensive simulation studies and it is applied in classification of the HIV+ and HIV- individuals.
]]></description>
<dc:creator>Brzyski, D.</dc:creator>
<dc:creator>Karas, M.</dc:creator>
<dc:creator>Ances, B.</dc:creator>
<dc:creator>Dzemidzic, M.</dc:creator>
<dc:creator>Goni, J.</dc:creator>
<dc:creator>Randolph, T. W.</dc:creator>
<dc:creator>Harezlak, J.</dc:creator>
<dc:date>2018-05-15</dc:date>
<dc:identifier>doi:10.1101/322420</dc:identifier>
<dc:title><![CDATA[Connectivity-Informed Adaptive Regularization for Generalized Outcomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/324269v1?rss=1">
<title>
<![CDATA[
Microscopy quantification of microbial birth and death dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/324269v1?rss=1</link>
<description><![CDATA[
Microbes live in dynamic environments where nutrient concentrations fluctuate. Quantifying fitness (birth and death) in a wide range of environments is critical for understanding microbial evolution as well as ecological interactions where one species alters the fitness of another. Here, using high-throughput time-lapse microscopy, we have quantified how Saccharomyces cerevisiae mutants incapable of synthesizing an essential metabolite grow or die in various concentrations of the required metabolite. We establish that cells normally expressing fluorescent proteins lose fluorescence upon death and that the total fluorescence in an imaging frame is proportional to the number of live cells even when cells form multiple layers. We validate our microscopy approach of measuring birth and death rates using flow cytometry, cell counting, and chemostat culturing. For lysine-requiring cells, very low concentrations of lysine are not detectably consumed and do not support cell birth, but delay the onset of death phase and reduce the death rate. In contrast, in low hypoxanthine, hypoxanthine-requiring cells can produce new cells, yet also die faster than in the absence of hypoxanthine. For both strains, birth rates under various metabolite concentrations are better described by the sigmoidal-shaped Moser model than the well-known Monod model, while death rates depend on the metabolite concentration and can vary with time. Our work reveals how time-lapse microscopy can be used to discover non-intuitive microbial dynamics and to quantify growth rates in many environments.
]]></description>
<dc:creator>Hart, S. F. M.</dc:creator>
<dc:creator>Skelding, D.</dc:creator>
<dc:creator>Waite, A. J.</dc:creator>
<dc:creator>Burton, J.</dc:creator>
<dc:creator>Xie, L.</dc:creator>
<dc:creator>Shou, W.</dc:creator>
<dc:date>2018-05-17</dc:date>
<dc:identifier>doi:10.1101/324269</dc:identifier>
<dc:title><![CDATA[Microscopy quantification of microbial birth and death dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/324707v1?rss=1">
<title>
<![CDATA[
Sera from individuals with narrowly focused influenza virus antibodies rapidly select viral escape mutations in ovo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/324707v1?rss=1</link>
<description><![CDATA[
Influenza viruses use distinct antibody escape mechanisms depending on the overall complexity of the antibody response that is encountered. When grown in the presence of a hemagglutinin (HA) monoclonal antibody, influenza viruses typically acquire a single HA mutation that reduces the binding of that specific monoclonal antibody. In contrast, when confronted with mixtures of HA monoclonal antibodies or polyclonal sera that have antibodies that bind several HA epitopes, influenza viruses acquire mutations that increase HA binding to host cells. Recent data from our laboratory and others suggest that some humans possess antibodies that are narrowly focused on HA epitopes that were present in influenza virus strains that they were likely exposed to in childhood. Here, we completed a series of experiments to determine if humans with narrowly focused HA antibody responses are able to select for influenza virus antigenic escape variants in ovo. We identified three human donors that possessed HA antibody responses that were heavily focused on a single HA antigenic site. Remarkably, sera from all three of these donors selected single HA escape mutations during in ovo passage experiments, similar to what has been previously reported for single monoclonal antibodies. These single HA mutations directly reduced binding of serum antibodies used for selection. We propose that new antigenic variants of influenza viruses might originate in individuals that produce antibodies that are narrowly focused on HA epitopes that were present in viral strains that they encountered in childhood.nnImportanceInfluenza vaccine strains must be updated frequently since circulating viral strains continuously change in antigenically important epitopes. Our previous studies have demonstrated that some individuals possess antibody responses that are narrowly focused on epitopes that were present in viral strains that they encountered during childhood. Here, we show that influenza viruses rapidly escape this type of polyclonal antibody response when grown in ovo by acquiring single mutations that directly prevent antibody binding. These studies improve our understanding of how influenza viruses evolve when confronted with narrowly focused polyclonal human antibodies.
]]></description>
<dc:creator>Davis, A. K. F.</dc:creator>
<dc:creator>McCormick, K.</dc:creator>
<dc:creator>Gumina, M. E.</dc:creator>
<dc:creator>Petrie, J.</dc:creator>
<dc:creator>Martin, E. T.</dc:creator>
<dc:creator>Xue, K. S.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Monto, A.</dc:creator>
<dc:creator>Bushman, F.</dc:creator>
<dc:creator>Hensley, S. E.</dc:creator>
<dc:date>2018-05-17</dc:date>
<dc:identifier>doi:10.1101/324707</dc:identifier>
<dc:title><![CDATA[Sera from individuals with narrowly focused influenza virus antibodies rapidly select viral escape mutations in ovo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/326421v1?rss=1">
<title>
<![CDATA[
ICeD-T Provides Accurate Estimates of Immune Cell Abundance in Tumor Samples by Allowing For Aberrant Gene Expression Patterns 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/326421v1?rss=1</link>
<description><![CDATA[
Immunotherapies have achieved phenomenal success in the treatment of cancer and promise even more breakthroughs in the near future. The need to understand the underlying mechanisms of immunotherapies and to develop precision immunotherapy regimens has spurred great interest in characterizing immune cell composition within the tumor microenvironment. Several methods have been developed to estimate immune cell composition using gene expression data from bulk tumor samples. However, these methods are not flexible enough to handle aberrant patterns of gene expression data, e.g., inconsistent cell type-specific gene expression between purified reference samples and this cell type in tumor samples. In this paper, we present a novel statistical model for expression deconvolution called ICeD-T (Immune Cell Deconvolution in Tumor tissues), which models gene expression by a log-normal distribution that is appropriate for both microarray and RNA-seq data. ICeD-T automatically identifies aberrant genes whose expressions are inconsistent with the deconvolution model and down-weights their contributions to cell type abundance estimates. We evaluated the performance of ICeD-T versus existing methods in simulation studies and several real data analyses. ICeD-T displayed comparable or superior performance to these competing methods. Applying these methods to assess the relationship between immunotherapy response and immune cell composition, ICeD-T is able to identify significant associations that are missed by its competitors.
]]></description>
<dc:creator>Wilson, D. R.</dc:creator>
<dc:creator>Ibrahim, J. G.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2018-05-21</dc:date>
<dc:identifier>doi:10.1101/326421</dc:identifier>
<dc:title><![CDATA[ICeD-T Provides Accurate Estimates of Immune Cell Abundance in Tumor Samples by Allowing For Aberrant Gene Expression Patterns]]></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/10.1101/326967v1?rss=1">
<title>
<![CDATA[
Comprehensive profiling of translation initiation in influenza virus infected cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/326967v1?rss=1</link>
<description><![CDATA[
Translation can initiate at alternate, non-canonical start codons in response to stressful stimuli in mammalian cells. Recent studies suggest that viral infection and anti-viral responses alter sites of translation initiation, and in some cases, lead to production of novel immune epitopes. Here we systematically investigate the extent and impact of alternate translation initiation in cells infected with influenza virus. We perform evolutionary analyses that suggest selection against non-canonical initiation at CUG codons in influenza virus lineages that have adapted to mammalian hosts. We then use ribosome profiling with the initiation inhibitor lactidomycin to experimentally delineate translation initiation sites in a human lung epithelial cell line infected with influenza virus. We identify several candidate sites of alternate initiation in influenza mRNAs, all of which occur at AUG codons that are downstream of canonical initiation codons. One of these candidate downstream start sites truncates 14 amino acids from the N-terminus of the N1 neuraminidase protein, resulting in loss of its cytoplasmic tail and a portion of the transmembrane domain. This truncated neuraminidase protein is expressed on the cell surface during influenza virus infection, is enzymatically active, and is conserved in most N1 viral lineages. Host transcripts induced by the anti-viral response are enriched for translation initiation at non-canonical start sites and non-AUG start codons. Together, our results systematically map the landscape of translation initiation during influenza virus infection, and shed light on the evolutionary forces shaping this landscape.
]]></description>
<dc:creator>Machkovech, H. M.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Subramaniam, A. R.</dc:creator>
<dc:date>2018-05-21</dc:date>
<dc:identifier>doi:10.1101/326967</dc:identifier>
<dc:title><![CDATA[Comprehensive profiling of translation initiation in influenza virus infected cells]]></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/10.1101/326983v1?rss=1">
<title>
<![CDATA[
RNA components of the spliceosome regulate tissue- and cancer-specific alternative splicing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/326983v1?rss=1</link>
<description><![CDATA[
Alternative splicing of pre-mRNAs plays a pivotal role during the establishment and maintenance of human cell types. Characterizing the trans-acting regulatory proteins that control alternative splicing in both healthy and malignant cells has therefore been the focus of much research. Recent work has established that even core protein components of the spliceosome, which are required for splicing to proceed, can nonetheless contribute to splicing regulation by modulating splice site choice. We here demonstrate that the RNA components of the spliceosome likewise influence alternative splicing decisions and contribute to the establishment of global splicing programs. Although these small nuclear RNAs (snRNAs), termed U1, U2, U4, U5, and U6 snRNA, are present in equal stoichiometry within the spliceosome, we found that their relative levels vary by an order of magnitude during development, across tissues, and between normal and malignant cells. Physiologically relevant perturbation of individual snRNAs drove widespread gene-specific differences in alternative splicing, but not transcriptome-wide splicing failure. Genes that were particularly sensitive to variations in snRNA abundance in a breast cancer cell line model were likewise preferentially mis-spliced within a clinically diverse cohort of invasive breast ductal carcinomas. As aberrant mRNA splicing is prevalent in many solid and liquid tumors, we propose that a full understanding of dysregulated pre-mRNA processing in cancers requires study of the RNA as well as protein components of the splicing machinery.
]]></description>
<dc:creator>Dvinge, H.</dc:creator>
<dc:creator>Guenthoer, J.</dc:creator>
<dc:creator>Porter, P. L.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:date>2018-05-21</dc:date>
<dc:identifier>doi:10.1101/326983</dc:identifier>
<dc:title><![CDATA[RNA components of the spliceosome regulate tissue- and cancer-specific alternative splicing]]></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/10.1101/329938v1?rss=1">
<title>
<![CDATA[
CD32+ and PD-1+ Lymph Node CD4 T Cells Support Persistent HIV-1 Transcription in Treated Aviremic Individuals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/329938v1?rss=1</link>
<description><![CDATA[
A recent study conducted in blood has proposed CD32 as the marker identifying the  elusive HIV reservoir. We have investigated the distribution of CD32+ CD4 T cells in blood and lymph nodes(LNs) of healthy HIV-1 uninfected, viremic untreated and long-term treated HIV-1 infected individuals and their relationship with PD-1+ CD4 T cells. The frequency of CD32+ CD4 T cells was increased in viremic as compared to treated individuals in LNs and a large proportion(up to 50%) of CD32+ cells co-expressed PD-1 and were enriched within T follicular helper cells(Tfh) cells. We next investigated the role of LN CD32+ CD4 T cells in the HIV reservoir. Total HIV DNA was enriched in CD32+ and PD-1+ CD4 T cells as compared to CD32- and PD-1- cells in both viremic and treated individuals but there was no difference between CD32+ and PD-1+ cells. There was not enrichment of latently infected cells with inducible HIV-1 in CD32+ versus PD-1+ cells in ART treated individuals. HIV-1 transcription was then analyzed in LN memory CD4 T cell populations sorted on the basis of CD32 and PD-1 expression. CD32+PD-1+ CD4 T cells were significantly enriched in cell associated HIV RNA as compared to CD32-PD-1-(average 5.2 fold in treated and 86.6 fold in viremics), to CD32+PD-1-(2.2 fold in treated and 4.3 fold in viremics) and to CD32-PD-1+ cell populations(2.2 fold in ART treated and 4.6 fold in viremics). Similar levels of HIV-1 transcription were found in CD32+PD-1- and CD32-PD-1+ CD4 T cells. Interestingly, the proportion of CD32+ and PD-1+ CD4 T cells negatively correlated with CD4 T cell counts and length of therapy while positively correlated with viremia. Therefore, the expression of CD32 identifies, independently of PD-1, a CD4 T cell population with persistent HIV-1 transcription and CD32 and PD-1 co-expression the CD4 T cell population with the highest levels of HIV-1 transcription in both viremic and treated individuals.nnImportanceThe existence of long-lived latently infected resting memory CD4 T cells represents a major obstacle to the eradication of HIV infection. Identifying cell markers defining latently infected cells containing replication competent virus is important in order to determine the mechanisms of HIV persistence and to develop novel therapeutic strategies to cure HIV infection. We provide evidence that PD-1 and CD32 may have a complementary role in better defining CD4 T cell populations infected with HIV-1. Furthermore, CD4 T cells co-expressing CD32 and PD-1 identify a CD4 T cell population with high levels of persistent HIV-1 transcription.
]]></description>
<dc:creator>Noto, A.</dc:creator>
<dc:creator>Procopio, F.</dc:creator>
<dc:creator>Banga, R.</dc:creator>
<dc:creator>Suffiotti, M.</dc:creator>
<dc:creator>Corpataux, J.-M.</dc:creator>
<dc:creator>Cavassini, M.</dc:creator>
<dc:creator>Fenwick, C.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Perreau, M.</dc:creator>
<dc:creator>Pantaleo, G.</dc:creator>
<dc:date>2018-05-24</dc:date>
<dc:identifier>doi:10.1101/329938</dc:identifier>
<dc:title><![CDATA[CD32+ and PD-1+ Lymph Node CD4 T Cells Support Persistent HIV-1 Transcription in Treated Aviremic Individuals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/330860v1?rss=1">
<title>
<![CDATA[
Ultrasensitive capture of human herpes simplex virus genomes directly from clinical samples reveals extraordinarily limited evolution in cell culture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/330860v1?rss=1</link>
<description><![CDATA[
Herpes simplex viruses (HSV) are difficult to sequence due to their large DNA genome, high GC content, and the presence of repeats. To date, most HSV genomes have been recovered from culture isolates, raising concern that these genomes may not accurately represent circulating clinical strains. We report the development and validation of a DNA oligonucleotide hybridization panel to recover near complete HSV genomes at abundances up to 50,000-fold lower than previously reported. Using copy number information on herpesvirus and host DNA background via quantitative PCR, we developed a protocol for pooling for cost-effective recovery of more than 50 HSV-1 or HSV-2 genomes per MiSeq run. We demonstrate the ability to recover >99% of the HSV genome at >100X coverage in 72 hours at viral loads that allow whole genome recovery from latently-infected ganglia. We also report a new computational pipeline for rapid HSV genome assembly and annotation. Using the above tools and a series of 17 HSV-1-positive clinical swabs sent to our laboratory for viral isolation, we show limited evolution of HSV-1 during viral isolation in human fibroblast cells compared to the original clinical samples. Our data indicate that previous studies using low passage clinical isolates of herpes simplex viruses are reflective of the viral sequences present in the lesion and thus can be used in phylogenetic analyses. We also detect superinfection within a single sample with unrelated HSV-1 strains recovered from separate oral lesions in an immunosuppressed patient during a 2.5-week period, illustrating the power of direct-from-specimen sequencing of HSV.nnImportanceHerpes simplex viruses affect more than 4 billion people across the globe, constituting a large burden of disease. Understanding global diversity of herpes simplex viruses is important for diagnostics and therapeutics as well as cure research and tracking transmission among humans. To date, most HSV genomics has been performed on culture isolates and DNA swabs with high quantities of virus. We describe the development of wet-lab and computational tools that enable the accurate sequencing of near-complete genomes of HSV-1 and HSV-2 directly from clinical specimens at abundances >50,000-fold lower than previously sequenced and at significantly reduced cost. We use these tools to profile circulating HSV-1 strains in the community and illustrate limited changes to the viral genome during the viral isolation process. These techniques enable cost-effective, rapid sequencing of HSV-1 and HSV-2 genomes that will help enable improved detection, surveillance, and control of this human pathogen.
]]></description>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Casto, A.</dc:creator>
<dc:creator>Cent, A.</dc:creator>
<dc:creator>Pepper, G.</dc:creator>
<dc:creator>Koelle, D. M.</dc:creator>
<dc:creator>Huang, M.-L.</dc:creator>
<dc:creator>Wald, A.</dc:creator>
<dc:creator>Johnston, C.</dc:creator>
<dc:creator>Jerome, K.</dc:creator>
<dc:date>2018-05-25</dc:date>
<dc:identifier>doi:10.1101/330860</dc:identifier>
<dc:title><![CDATA[Ultrasensitive capture of human herpes simplex virus genomes directly from clinical samples reveals extraordinarily limited evolution in cell culture]]></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/10.1101/334201v1?rss=1">
<title>
<![CDATA[
Characterization of influenza B virus variants with reduced neuraminidase inhibitor susceptibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/334201v1?rss=1</link>
<description><![CDATA[
Treatment options for influenza B virus infections are limited to neuraminidase inhibitors (NAIs) which block the neuraminidase (NA) glycoprotein on the virion surface. The development of NAI resistance would therefore result in a loss of antiviral treatment options for influenza B infections. This study characterized two contemporary influenza B viruses with known resistance-conferring NA amino acid substitutions, D197N and H273Y, detected during routine surveillance. The D197N and H273Y variants were characterized in vitro by assessing NA enzyme activity and affinity, as well as replication in cell culture compared to NAI-sensitive wild-type viruses. In vivo studies were also performed in ferrets to assess the replication and transmissibility of each variant. Mathematical models were used to analyse within-host and between-host fitness of variants relative to wild-type viruses. The data revealed that the H273Y variant had similar NA enzyme function relative to its wild-type but had slightly reduced replication and transmission efficiency in vivo. The D197N variant had impaired NA enzyme function but there was no evidence of reduction in replication or transmission efficiency in ferrets. Our data suggest that the influenza B variant with H273Y NA substitution had a more notable reduction in fitness compared to wild-type viruses than the influenza B variant with the D197N NA substitution. Although a D197N variant is yet to become widespread, it is the most commonly detected NAI-resistant influenza B virus in surveillance studies. Our results highlight the need to carefully monitor circulating viruses for the spread of influenza B viruses with the D197N NA substitution.
]]></description>
<dc:creator>Farrukee, R.</dc:creator>
<dc:creator>Alex, Z. E.</dc:creator>
<dc:creator>James, M.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Reading, P.</dc:creator>
<dc:creator>Hurt, A. C.</dc:creator>
<dc:date>2018-05-30</dc:date>
<dc:identifier>doi:10.1101/334201</dc:identifier>
<dc:title><![CDATA[Characterization of influenza B virus variants with reduced neuraminidase inhibitor susceptibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/335554v1?rss=1">
<title>
<![CDATA[
cytometree: a binary tree algorithm for automatic gating in cytometry analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/335554v1?rss=1</link>
<description><![CDATA[
MotivationFlow cytometry is a powerful technology that allows the high-throughput quantification of dozens of surface and intracellular proteins at the single-cell level. It has become the most widely used technology for immunophenotyping of cells over the past three decades. Due to the increasing complexity of cytometry experiments (more cells and more markers), traditional manual flow cytometry data analysis has become untenable due to its subjectivity and time-consuming nature.nnResultsWe present a new unsupervised algorithm called "cytometree" to perform automated population discovery (aka gating) in flow cytometry. cytometree is based on the construction of a binary tree, the nodes of which are subpopulations of cells. At each node, the marker distributions are modeled by mixtures of normal distribution. Node splitting is done according to a normalized difference of Akaike information criteria (AIC) between the two models. Post-processing of the tree structure and derived populations allows us to complete the annotation of the derived populations. The algorithm is shown to perform better than the state-of-the-art unsupervised algorithms previously proposed on panels introduced by the Flow Cytometry: Critical Assessment of Population Identification Methods (FlowCAP I) project. The algorithm is also applied to a T-cell panel proposed by the Human Immunology Project Consortium (HIPC) program; it also outperforms the best unsupervised open-source available algorithm while requiring the shortest computation time.nnAvailabilityAn R package named "cytometree" is available on the CRAN repository.nnContactdaniel.commenges@u-bordeaux.fr; rodolphe.thiebaut@u-bordeaux.frnnSupplementary informationSupplementary data are available.
]]></description>
<dc:creator>Commenges, D.</dc:creator>
<dc:creator>Alkhassim, C.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Hejblum, B.</dc:creator>
<dc:creator>Thiebaut, R.</dc:creator>
<dc:date>2018-05-31</dc:date>
<dc:identifier>doi:10.1101/335554</dc:identifier>
<dc:title><![CDATA[cytometree: a binary tree algorithm for automatic gating in cytometry analysis]]></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/10.1101/340653v1?rss=1">
<title>
<![CDATA[
Bayesian Classification of Microbial Communities Based on 16S rRNA Metagenomic Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/340653v1?rss=1</link>
<description><![CDATA[
We propose a Bayesian method for the classification of 16S rRNA metagenomic profiles of bacterial abundance, by introducing a Poisson-Dirichlet-Multinomial hierarchical model for the sequencing data, constructing a prior distribution from sample data, calculating the posterior distribution in closed form; and deriving an Optimal Bayesian Classifier (OBC). The proposed algorithm is compared to state-of-the-art classification methods for 16S rRNA metagenomic data, including Random Forests and the phylogeny-based Metaphyl algorithm, for varying sample size, classification difficulty, and dimensionality (number of OTUs), using both synthetic and real metagenomic data sets. The results demonstrate that the proposed OBC method, with either noninformative or constructed priors, is competitive or superior to the other methods. In particular, in the case where the ratio of sample size to dimensionality is small, it was observed that the proposed method can vastly outperform the others.nnAuthor summaryRecent studies have highlighted the interplay between host genetics, gut microbes, and colorectal tumor initiation/progression. The characterization of microbial communities using metagenomic profiling has therefore received renewed interest. In this paper, we propose a method for classification, i.e., prediction of different outcomes, based on 16S rRNA metagenomic data. The proposed method employs a Bayesian approach, which is suitable for data sets with small ration of number of available instances to the dimensionality. Results using both synthetic and real metagenomic data show that the proposed method can outperform other state-of-the-art metagenomic classification algorithms.
]]></description>
<dc:creator>Bahadorinejad, A.</dc:creator>
<dc:creator>Ivanov, I.</dc:creator>
<dc:creator>Lampe, J.</dc:creator>
<dc:creator>Hullar, M.</dc:creator>
<dc:creator>Chapkin, R.</dc:creator>
<dc:creator>Braga-Neto, U.</dc:creator>
<dc:date>2018-06-06</dc:date>
<dc:identifier>doi:10.1101/340653</dc:identifier>
<dc:title><![CDATA[Bayesian Classification of Microbial Communities Based on 16S rRNA Metagenomic Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/342907v1?rss=1">
<title>
<![CDATA[
DataPackageR: Reproducible data preprocessing, standardization and sharing using R/Bioconductor for collaborative data analysis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/342907v1?rss=1</link>
<description><![CDATA[
A central tenet of reproducible research is that scientific results are published along with the underlying data and software code necessary to reproduce and verify the findings. A host of tools and software have been released that facilitate such work-flows and scientific journals have increasingly demanded that code and primary data be made available with publications. There has been little practical advice on implementing reproducible research work-flows for large  omics or systems biology data sets used by teams of analysts working in collaboration. In such instances it is important to ensure all analysts use the same version of a data set for their analyses. Yet, instantiating relational databases and standard operating procedures can be unwieldy, with high "startup" costs and poor adherence to procedures when they deviate substantially from an analysts usual work-flow. Ideally a reproducible research work-flow should fit naturally into an individuals existing work-flow, with minimal disruption. Here, we provide an overview of how we have leveraged popular open source tools, including Bioconductor, Rmarkdown, git version control, R, and specifically Rs package system combined with a new tool DataPackageR, to implement a lightweight reproducible research work-flow for preprocessing large data sets, suitable for sharing among small-to-medium sized teams of computational scientists. Our primary contribution is the DataPackageR tool, which decouples time-consuming data processing from data analysis while leaving a traceable record of how raw data is processed into analysis-ready data sets. The software ensures packaged data objects are properly documented and performs checksum verification of these along with basic package version management, and importantly, leaves a record of data processing code in the form of package vignettes. Our group has implemented this work-flow to manage, analyze and report on pre-clinical immunological trial data from multi-center, multi-assay studies for the past three years.
]]></description>
<dc:creator>Finak, G.</dc:creator>
<dc:creator>Mayer, B.</dc:creator>
<dc:creator>Fulp, W.</dc:creator>
<dc:creator>Obrecht, P.</dc:creator>
<dc:creator>Sato, A.</dc:creator>
<dc:creator>Chung, E.</dc:creator>
<dc:creator>Holman, D.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:date>2018-06-08</dc:date>
<dc:identifier>doi:10.1101/342907</dc:identifier>
<dc:title><![CDATA[DataPackageR: Reproducible data preprocessing, standardization and sharing using R/Bioconductor for collaborative data analysis.]]></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/10.1101/358481v1?rss=1">
<title>
<![CDATA[
Microbial coexistence through chemical-mediated interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/358481v1?rss=1</link>
<description><![CDATA[
Many microbial functions happen within communities of interacting species. Explaining how species with intrinsically disparate fitness can coexist is important for applications such as manipulating host-associated microbiota or engineering industrial communities. Previous coexistence studies have often neglected interaction mechanisms. Here, we formulate and experimentally constrain a model in which chemical mediators of microbial interactions (e.g. metabolites or waste-products) are explicitly incorporated. We construct many instances of coexistence by simulating community assembly through enrichment and ask how species interactions can explain coexistence. We show that growth-facilitating influences between members are favored in assembled communities. Among negative influences, self-restraint, such as production of self-inhibiting waste, contributes to coexistence, whereas inhibition of other species disrupts coexistence. Coexistence is also favored when interactions are mediated by depletable chemicals that get consumed or degraded, rather than by reusable chemicals that are unaffected by recipients. Our model creates null predictions for coexistence driven by chemical-mediated interactions.
]]></description>
<dc:creator>Niehaus, L.</dc:creator>
<dc:creator>Boland, I.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Fu, D.</dc:creator>
<dc:creator>Henckel, C.</dc:creator>
<dc:creator>Chaung, K.</dc:creator>
<dc:creator>Espinoza Miranda, S.</dc:creator>
<dc:creator>Dyckman, S.</dc:creator>
<dc:creator>Crum, M.</dc:creator>
<dc:creator>Dedrick, S.</dc:creator>
<dc:creator>Shou, W.</dc:creator>
<dc:creator>Momeni, B.</dc:creator>
<dc:date>2018-06-29</dc:date>
<dc:identifier>doi:10.1101/358481</dc:identifier>
<dc:title><![CDATA[Microbial coexistence through chemical-mediated interactions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/363432v1?rss=1">
<title>
<![CDATA[
A Virus-Packageable CRISPR Screen Identifies Host Factors Mediating Interferon Inhibition of HIV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/363432v1?rss=1</link>
<description><![CDATA[
Interferon (IFN) inhibits HIV replication by inducing an array of antiviral effectors. Here we describe a novel CRISPR knockout screening approach to identify the ensemble of these HIV restriction factors. We assembled a CRISPR sgRNA library specific for Interferon Stimulated Genes (ISGs) into a modified lentiviral vector that allows for packaging of sgRNA-encoding genomes in trans into budding HIV-1 particles. We observed that knockout of Zinc Antiviral Protein (ZAP) improved the performance of the screen due to ZAP-mediated inhibition of the vector. We identify a small panel of IFN-induced HIV restriction factors, including MxB, IFITM1, Tetherin/BST2 and TRIM5 which together explain the inhibitory effects of IFN on the HIV-1 LAI strain in THP-1 cells. Further, we identify novel HIV dependency factors, including SEC62 and TLR2. The ability of IFN-induced restriction factors to inhibit an HIV strain to replicate in human cells suggests that these human restriction factors are incompletely antagonized.
]]></description>
<dc:creator>Ohainle, M.</dc:creator>
<dc:creator>Pendergast, L.</dc:creator>
<dc:creator>Vermeire, J.</dc:creator>
<dc:creator>Roesch, F.</dc:creator>
<dc:creator>Humes, D.</dc:creator>
<dc:creator>Basom, R.</dc:creator>
<dc:creator>Delrow, J. J.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2018-07-20</dc:date>
<dc:identifier>doi:10.1101/363432</dc:identifier>
<dc:title><![CDATA[A Virus-Packageable CRISPR Screen Identifies Host Factors Mediating Interferon Inhibition of HIV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/364430v1?rss=1">
<title>
<![CDATA[
Reconciling disparate estimates of viral genetic diversity during human influenza infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/364430v1?rss=1</link>
<description><![CDATA[
Deep sequencing can measure viral genetic diversity within human influenza infections, but published studies disagree in their estimates of how much genetic diversity is typically present. One large-scale deep-sequencing study of human influenza reported high levels of shared viral genetic diversity among infected individuals in Hong Kong, but subsequent studies of other cohorts have reported little shared viral diversity. We re-analyze sequencing data from four studies of within-host genetic diversity encompassing more than 500 acute human influenza infections. We identify an anomaly in the Hong Kong data that provides a technical explanation for these discrepancies: read pairs from this study are often split between different biological samples, indicating that some reads are incorrectly assigned. These technical abnormalities explain the high levels of within-host variation and loose transmission bottlenecks reported by this study. Studies without these anomalies consistently report low levels of genetic diversity in acute human influenza infections.
]]></description>
<dc:creator>Xue, K. S.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2018-07-08</dc:date>
<dc:identifier>doi:10.1101/364430</dc:identifier>
<dc:title><![CDATA[Reconciling disparate estimates of viral genetic diversity during human influenza infections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/381897v1?rss=1">
<title>
<![CDATA[
An assay for de novo kinetochore assembly reveals a key role for the CENP-T pathway in budding yeast 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/381897v1?rss=1</link>
<description><![CDATA[
Chromosome segregation depends on the kinetochore, the machine that establishes force-bearing attachments between DNA and spindle microtubules. Kinetochores are formed every cell cycle via a highly regulated process that requires coordinated assembly of multiple subcomplexes on specialized chromatin. To elucidate the underlying mechanisms, we developed an assay to assemble kinetochores de novo using centromeric DNA and budding yeast extracts. Assembly is enhanced by mitotic phosphorylation of the Dsn1 kinetochore protein and generates kinetochores capable of binding microtubules. We used this assay to investigate why kinetochores recruit the microtubule-binding Ndc80 complex via two receptors: the Mis12 complex and CENP-T. Although the CENP-T pathway is non-essential in yeast, we demonstrate that it becomes essential for viability and Ndc80c recruitment when the Mis12 pathway is crippled by defects in Dsn1 phosphorylation. Assembling kinetochores de novo in yeast extracts provides a powerful and genetically tractable method to elucidate critical regulatory events in the future.
]]></description>
<dc:creator>Lang, J.</dc:creator>
<dc:creator>Barber, A.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2018-08-07</dc:date>
<dc:identifier>doi:10.1101/381897</dc:identifier>
<dc:title><![CDATA[An assay for de novo kinetochore assembly reveals a key role for the CENP-T pathway in budding yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/383018v1?rss=1">
<title>
<![CDATA[
L. pneumophila deploys a self-active inhibitor for inter-Legionella competition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/383018v1?rss=1</link>
<description><![CDATA[
To persist in the extracellular state, the bacterial pathogen Legionella pneumophila must withstand competition from neighboring bacteria. Here, we find that L. pneumophila can antagonize the growth of neighboring Legionella species using a secreted inhibitor: HGA (homogentisic acid), the unstable, redox-active precursor molecule to L. pneumophilas brown-black pigment. Unexpectedly, we find that L. pneumophila can itself be inhibited by HGA secreted from neighboring, isogenic strains. Our genetic approaches further identify lpg1681 as a gene that modulates L. pneumophila susceptibility to HGA. We find that L. pneumophila sensitivity to HGA is density-dependent and cell intrinsic. This resistance is not mediated by the stringent response nor the previously described Legionella quorum-sensing pathway. Instead, we find that L. pneumophila cells secrete HGA only when they are conditionally HGA-resistant, which allows these bacteria to produce a potentially self-toxic molecule while restricting the opportunity for self-harm. We speculate that established Legionella communities may deploy molecules such as HGA as an unusual public good that can protect against invasion by low-density competitors.
]]></description>
<dc:creator>Levin, T. C.</dc:creator>
<dc:creator>Goldspiel, B. P.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2018-08-02</dc:date>
<dc:identifier>doi:10.1101/383018</dc:identifier>
<dc:title><![CDATA[L. pneumophila deploys a self-active inhibitor for inter-Legionella competition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/397679v1?rss=1">
<title>
<![CDATA[
RNA sequencing of the in vivo human herpesvirus 6B transcriptome to identify targets for clinical assays distinguishing between latent and active infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/397679v1?rss=1</link>
<description><![CDATA[
Human herpesvirus 6B (HHV-6B) DNA is frequently detected in human samples, especially after hematopoietic cell transplantation (HCT). Diagnostic assays distinguishing HHV-6B reactivation from latency are limited, and this has contributed to confusion in research and made the design of clinical approaches to diagnose and treat HHV-6-associated diseases challenging. We used RNA sequencing to characterize and compare the HHV-6B transcriptome in multiple in vivo and in vitro sample types, including 1) whole blood from HCT recipients with and without HHV-6B plasma viremia; 2) tumor tissue samples from subjects with large B cell lymphoma infected with HHV-6B; 3) lymphoblastoid cell lines from subjects with inherited chromosomally integrated HHV-6B or latent infection with HHV-6B; and 4) HHV-6B Z29 infected SupT1 CD4+ T cells. We demonstrated substantial overlap in the HHV-6B transcriptome observed in in vivo and in vitro samples, although there was variability in the breadth and quantity of gene expression across samples. No HHV-6B transcripts were detected in whole blood samples from subjects without plasma HHV-6B viremia. The HHV-6B viral polymerase gene U38 was the only HHV-6B transcript detected in all RNA-seq data sets and was one of the most highly expressed genes. Using a novel reverse transcription PCR assay targeting HHV-6B U38, we identified U38 messenger RNA in all tested whole blood samples from patients with concurrent HHV-6B viremia, indicating its utility as a diagnostic assay for HHV-6B replication. This study demonstrates the feasibility of pathogen transcriptome analyses in HCT recipients to identify better targets for diagnostic, and potentially therapeutic, applications.nnIMPORTANCEInfection with human herpesvirus 6B (HHV-6B), a DNA virus, occurs early in life, results in chronic viral latency in diverse cell types, and affects the population at large. Additionally, HHV- 6B can integrate into germline chromosomes, resulting in individuals with viral DNA in every nucleated cell. Given that PCR to detect viral DNA is the mainstay for diagnosing HHV-6B infection, the characteristics of HHV-6B infection complicate efforts to distinguish between latent and active viral infection, particularly in immunocompromised patients who have frequent HHV- 6B reactivation. In this study, we used RNA sequencing to characterize the HHV-6B gene expression profile in multiple sample types, and our findings identified evidence-based targets for diagnostic tests that distinguish between latent and active viral infection.
]]></description>
<dc:creator>Hill, J. A.</dc:creator>
<dc:creator>Ikoma, M.</dc:creator>
<dc:creator>Zerr, D. M.</dc:creator>
<dc:creator>Basom, R. S.</dc:creator>
<dc:creator>Peddu, V.</dc:creator>
<dc:creator>Huang, M.-L.</dc:creator>
<dc:creator>Hall Sedlak, R.</dc:creator>
<dc:creator>Jerome, K.</dc:creator>
<dc:creator>Boeckh, M.</dc:creator>
<dc:creator>Barcy, S.</dc:creator>
<dc:date>2018-08-22</dc:date>
<dc:identifier>doi:10.1101/397679</dc:identifier>
<dc:title><![CDATA[RNA sequencing of the in vivo human herpesvirus 6B transcriptome to identify targets for clinical assays distinguishing between latent and active infections]]></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/10.1101/402743v1?rss=1">
<title>
<![CDATA[
A natural history model for planning prostate cancer testing: calibration and validation using Swedish registry data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/402743v1?rss=1</link>
<description><![CDATA[
Recent prostate cancer screening trials have given conflicting results and it is unclear how to reduce prostate cancer mortality while minimising overdiagnosis and overtreatment. Prostate cancer testing is a partially observable process, and planning for testing requires either extrapolation from randomised controlled trials or, more flexibly, modelling of the cancer natural history.nnAn existing US prostate cancer natural history model (Gulati et al, Biostatistics 2010;11:707-719) did not model for differences in survival between Gleason 6 and 7 cancers and predicted too few Gleason 7 cancers for contemporary Sweden. We re-implemented and re-calibrated the US model to Sweden. We extended the model to more finely describe the disease states, their time to biopsy-detectable cancer and prostate cancer survival. We first calibrated the model to the incidence rate ratio observed in the European Randomised Study of Screening for Prostate Cancer (ERSPC) together with age-specific cancer staging observed in the Stockholm PSA (prostate-specific antigen) and Biopsy Register; we then calibrated age-specific survival by disease states under contemporary testing and treatment using the Swedish National Prostate Cancer Register.nnAfter calibration, we were able to closely match observed prostate cancer incidence trends in Sweden. Assuming that patients detected at an earlier stage by screening receive a commensurate survival improvement, we find that the calibrated model replicates the observed mortality reduction in a simulation of ERSPC.nnUsing the resulting model, we predicted incidence and mortality following the introduction of regular testing. Compared with a model of the current testing pattern, organised 8 yearly testing for men aged 55-69 years was predicted to reduce prostate cancer incidence by 0.11% with no increase in the mortality rate. The model is open source and suitable for planning for effective prostate cancer screening into the future.nnAuthor summaryA naive perspective is that cancer screening is simple: people are screened, some cancers are detected early, and cancer mortality rates decline. However, the mathematics for screening becomes difficult quickly, it is hard to infer causation from observational data, and even large randomised screening studies provide limited evidence. Simulations are therefore important for planning cancer screening.nnWe found an older US prostate cancer natural history model to be poorly suited for contemporary Sweden. We therefore re-implemented and re-calibrated the US model using data from Swedish registries.nnOur revised model, the Stockholm "Prostata" model, provides predictions similar to those observed in the detailed Swedish registers on prostate cancer incidence and mortality. By modelling the mechanisms of the screening effect, we can predict the benefits and harms under a range of screening interventions.
]]></description>
<dc:creator>Karlsson, A.</dc:creator>
<dc:creator>Jauhiainen, A.</dc:creator>
<dc:creator>Gulati, R.</dc:creator>
<dc:creator>Eklund, M.</dc:creator>
<dc:creator>Grönberg, H.</dc:creator>
<dc:creator>Etzioni, R.</dc:creator>
<dc:creator>Clements, M.</dc:creator>
<dc:date>2018-08-28</dc:date>
<dc:identifier>doi:10.1101/402743</dc:identifier>
<dc:title><![CDATA[A natural history model for planning prostate cancer testing: calibration and validation using Swedish registry data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/406355v1?rss=1">
<title>
<![CDATA[
An antigenic atlas of HIV-1 escape from broadly neutralizing antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/406355v1?rss=1</link>
<description><![CDATA[
Anti-HIV broadly neutralizing antibodies (bnAbs) have revealed vaccine targets on the viruss Env protein and are themselves promising immunotherapeutics. The efficacy of bnAb-based therapies and vaccines depends in part on how readily the virus can escape neutralization. While structural studies can define contacts between bnAbs and Env, only functional studies can define mutations that confer escape. Here we map how all single amino-acid mutations to Env affect neutralization of HIV by nine bnAbs targeting five epitopes. For most bnAbs, mutations at only a small fraction of structurally defined contact sites mediated escape, and most escape occurred at sites that are near but do not directly contact the antibody. The mutations selected by two pooled bnAbs were similar to those expected from the combination of the bnAbs independent action. Overall, our mutation-level antigenic atlas provides a comprehensive dataset for understanding viral immune escape and refining therapies and vaccines.
]]></description>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Arenz, D.</dc:creator>
<dc:creator>Weight, H.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2018-08-31</dc:date>
<dc:identifier>doi:10.1101/406355</dc:identifier>
<dc:title><![CDATA[An antigenic atlas of HIV-1 escape from broadly neutralizing antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/414995v1?rss=1">
<title>
<![CDATA[
Limited marginal utility of deep sequencing for HIV drug resistance testing in the age of integrase inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/414995v1?rss=1</link>
<description><![CDATA[
HIV drug resistance genotyping is a critical tool in the clinical management of HIV infections. Although resistance genotyping has traditionally been conducted using Sanger sequencing, next-generation sequencing (NGS) is emerging as a powerful tool due to its ability to detect lower frequency alleles. However, the value added from NGS approaches to antiviral resistance testing remains to be demonstrated. We compared the variant detection capacity of NGS versus Sanger sequencing methods for resistance genotyping of 144 drug resistance tests (105 protease-reverse transcriptase tests and 39 integrase tests) submitted to our clinical virology laboratory over a four-month period in 2016 for Sanger-based HIV drug resistance testing. NGS detected all true high frequency drug resistance mutations (>20% frequency) found by Sanger sequencing, with greater accuracy in one instance of a Sanger-detected false positive. Freely available online NGS variant callers Hydra and PASeq were superior to Sanger methods for interpretations of allele linkage and automated variant calling. NGS additionally detected low frequency mutations (1-20% frequency) associated with higher levels of drug resistance in 30/105 (29%) of protease-reverse transcriptase tests and 4/39 (10%) of integrase tests. Clinical follow-up of 69 individuals for a median of 674 days found no difference in rates of virological failure between individuals with and without low frequency mutations, although rates of virological failure were higher for individuals with drug-relevant low frequency mutations. However, all 27 individuals who experienced virological failure reported poor adherence to their drug regimen during preceding follow-up time, and all 19 who subsequently improved their adherence achieved viral suppression at later time points consistent with a lack of clinical resistance. In conclusion, in a population with low antiviral resistance emergence, NGS methods detected numerous instances of minor alleles that did not result in subsequent bona fide virological failure due to antiviral resistance.nnImportanceGenotypic antiviral resistance testing for HIV is an essential component of the clinical microbiology and virology laboratory. Next-generation sequencing (NGS) has emerged as a powerful tool for the detection of low frequency sequence variants (allele frequencies <20%). Whether detecting these low frequency mutations in HIV contributes to improved patient health, however, remains unclear. We compared NGS to conventional Sanger sequencing for detecting resistance mutations for 144 HIV drug resistance tests submitted to our clinical virology laboratory and detected low frequency mutations in 24% of tests. Over approximately two years of follow-up for 69 patients for which we had access to electronic health records, no patients had virological failure due to antiviral resistance. Instead, virological failure was entirely explained by medication non-adherence. While larger studies are required, we suggest that detection of low frequency variants by NGS presents limited marginal clinical utility when compared to standard of care.
]]></description>
<dc:creator>Dalmat, R. R.</dc:creator>
<dc:creator>Makhsous, N.</dc:creator>
<dc:creator>Pepper, G.</dc:creator>
<dc:creator>Magaret, A.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Wald, A.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:date>2018-09-12</dc:date>
<dc:identifier>doi:10.1101/414995</dc:identifier>
<dc:title><![CDATA[Limited marginal utility of deep sequencing for HIV drug resistance testing in the age of integrase inhibitors]]></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/10.1101/416032v1?rss=1">
<title>
<![CDATA[
Rapid development of an infant-derived HIV-1 broadly neutralizing antibody lineage 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/416032v1?rss=1</link>
<description><![CDATA[
HIV-infected infants develop broadly neutralizing plasma responses with more rapid kinetics than adults, suggesting the ontogeny of infant responses could better inform a path to achievable vaccine targets. We developed computational methods to reconstruct the developmental lineage of BF520.1, the first example of a HIV-specific broadly neutralizing antibody (bnAb) from an infant. The BF520.1 inferred naive precursor binds HIV Env and a bnAb evolved within six months of infection and required only 3% mutation. Mutagenesis and structural analyses revealed that for this infant bnAb, substitutions in the kappa chain were critical for activity, particularly in CDRL1. Overall, the developmental pathway of this infant antibody includes features distinct from adult antibodies, including several that may be amenable to better vaccine responses.
]]></description>
<dc:creator>Simonich, C. A.</dc:creator>
<dc:creator>Doepker, L.</dc:creator>
<dc:creator>Ralph, D.</dc:creator>
<dc:creator>Williams, J. A.</dc:creator>
<dc:creator>Dhar, A.</dc:creator>
<dc:creator>Yaffe, Z.</dc:creator>
<dc:creator>Gentles, L.</dc:creator>
<dc:creator>Small, C. T.</dc:creator>
<dc:creator>Oliver, B.</dc:creator>
<dc:creator>Vigdorovich, V.</dc:creator>
<dc:creator>Mangala Prasad, V.</dc:creator>
<dc:creator>Nduati, R.</dc:creator>
<dc:creator>Sather, D. N.</dc:creator>
<dc:creator>Lee, K. K.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2018-09-13</dc:date>
<dc:identifier>doi:10.1101/416032</dc:identifier>
<dc:title><![CDATA[Rapid development of an infant-derived HIV-1 broadly neutralizing antibody lineage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/417790v1?rss=1">
<title>
<![CDATA[
Quantitative proteomics reveals key roles for post-transcriptional gene regulation in the molecular pathology of FSHD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/417790v1?rss=1</link>
<description><![CDATA[
DUX4 is a transcription factor whose misexpression in skeletal muscle causes facioscapulohumeral muscular dystrophy (FSHD). While DUX4s transcriptional activity has been extensively characterized, the DUX4-induced proteome remains undescribed. Here, we report concurrent measurement of RNA and protein levels in DUX4-expressing cells via RNA-seq and quantitative mass spectrometry. DUX4 transcriptional targets were robustly translated, confirming the likely clinical relevance of proposed FSHD biomarkers. However, a multitude of mRNAs and proteins exhibited discordant expression changes upon DUX4 expression. Our dataset revealed unexpected proteomic, but not transcriptomic, dysregulation of diverse molecular pathways, including Golgi apparatus fragmentation, as well as extensive post-transcriptional buffering of stress response genes. Key components of RNA degradation machineries, including UPF1, UPF3B, and XRN1, exhibited suppressed protein, but not mRNA, levels, explaining the build-up of aberrant RNAs that characterizes DUX4-expressing cells. Our results provide a resource for the FSHD community and illustrate the importance of post-transcriptional processes to DUX4-induced pathology.
]]></description>
<dc:creator>Jagannathan, S.</dc:creator>
<dc:creator>Ogata, Y.</dc:creator>
<dc:creator>Gafken, P.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:date>2018-09-14</dc:date>
<dc:identifier>doi:10.1101/417790</dc:identifier>
<dc:title><![CDATA[Quantitative proteomics reveals key roles for post-transcriptional gene regulation in the molecular pathology of FSHD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/420273v1?rss=1">
<title>
<![CDATA[
Trans-ethnic genome-wide association study provides insight into effector genes and molecular mechanisms for kidney function and highlights a causal effect on kidney-specific disease aetiologies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/420273v1?rss=1</link>
<description><![CDATA[
Chronic kidney disease (CKD) affects [~]10% of the global population, with considerable ethnic differences in prevalence and aetiology. We assembled genome-wide association studies (GWAS)1-3 of estimated glomerular filtration rate (eGFR), a measure of kidney function that defines CKD, in 312,468 individuals from four ancestry groups. We identified 93 loci (20 novel), which were delineated to 127 distinct association signals. These signals were homogenous across ancestries, and were enriched for protein-coding exons, kidney-specific histone modifications, and transcription factor binding sites for HDAC2 and EZH2. Fine-mapping revealed 40 high-confidence variants driving eGFR associations and highlighted potential causal genes with cell-type specific expression in glomerulus, and proximal and distal nephron. Mendelian randomisation (MR) supported causal effects of eGFR on overall and cause-specific CKD, kidney stone formation, diastolic blood pressure (DBP) and hypertension. These results define novel molecular mechanisms and effector genes for eGFR, offering insight into clinical outcomes and routes to CKD treatment development.
]]></description>
<dc:creator>Morris, A. P.</dc:creator>
<dc:creator>Le, T. H.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Akbarov, A.</dc:creator>
<dc:creator>van der Most, P. P.</dc:creator>
<dc:creator>Mahajan, A.</dc:creator>
<dc:creator>Hemani, G.</dc:creator>
<dc:creator>Gaulton, K. J.</dc:creator>
<dc:creator>Nadkarni, G.</dc:creator>
<dc:creator>Valladares-Salgado, A.</dc:creator>
<dc:creator>Wacher-Rodarte, N.</dc:creator>
<dc:creator>Mychaleckyj, J. C.</dc:creator>
<dc:creator>Dueker, N. D.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Hai, Y.</dc:creator>
<dc:creator>Haessler, J.</dc:creator>
<dc:creator>Kamatani, Y.</dc:creator>
<dc:creator>Stilp, A.</dc:creator>
<dc:creator>Zhu, G.</dc:creator>
<dc:creator>Cook, J. P.</dc:creator>
<dc:creator>Arnlov, J.</dc:creator>
<dc:creator>Blanton, S. H.</dc:creator>
<dc:creator>de Borst, M.</dc:creator>
<dc:creator>Bottinger, E. P.</dc:creator>
<dc:creator>Buchanan, T. A.</dc:creator>
<dc:creator>Charchar, F. J.</dc:creator>
<dc:creator>Damman, J.</dc:creator>
<dc:creator>Eales, J. M.</dc:creator>
<dc:creator>Gharavi, A. G.</dc:creator>
<dc:creator>Giedraitis, V.</dc:creator>
<dc:creator>Heath, A. C.</dc:creator>
<dc:creator>Ipp, E.</dc:creator>
<dc:creator>Kiryluk, K.</dc:creator>
<dc:creator>Kubo, M.</dc:creator>
<dc:creator>Larsson, A.</dc:creator>
<dc:creator>Lindgren, C.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Madden, P. A.</dc:creator>
<dc:creator>Kramer, H. J.</dc:creator>
<dc:creator>Montgomery, G. W.</dc:creator>
<dc:creator>Papanicolaou, G. J.</dc:creator>
<dc:creator>Raffel, L. J.</dc:creator>
<dc:creator>S</dc:creator>
<dc:date>2018-09-18</dc:date>
<dc:identifier>doi:10.1101/420273</dc:identifier>
<dc:title><![CDATA[Trans-ethnic genome-wide association study provides insight into effector genes and molecular mechanisms for kidney function and highlights a causal effect on kidney-specific disease aetiologies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/424275v1?rss=1">
<title>
<![CDATA[
Successes and failures of the live-attenuated influenza vaccine, can we do better? 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/424275v1?rss=1</link>
<description><![CDATA[
Live-attenuated vaccines are usually highly effective against many acute viral infections. However, the effectiveness of the live attenuated influenza vaccine (LAIV) can vary widely, ranging from 0% effectiveness in some studies done in the United States to 50% in studies done in Europe. The reasons for these discrepancies remain largely unclear. In this paper we use mathematical models to explore how the efficacy of LAIV is affected by the degree of mismatch with the currently circulating influenza strain and interference with pre-existing immunity. The model incorporates two key antigenic distances - the distance between pre-existing immunity and the currently circulating strain as well as the LAIV strain. Our models show that a LAIV that is matched with the currently circulating strain is likely to have only modest efficacy. Our results suggest that the efficacy of the vaccine would be increased (optimized) if, rather than being matched to the circulating strain, it is antigenically slightly further from pre-existing immunity compared with the circulating strain. The models also suggest two regimes in which LAIV that is matched to circulating strains may provide effective protection. The first is in children before they have built immunity from circulating strains. The second is in response to novel strains (such as antigenic shifts) which are at substantial antigenic distance from previously circulating strains. Our models provide an explanation for the variation in vaccine effectiveness, both between children and adults as well as between studies of vaccine effectiveness observed during the 2014-15 influenza season in different countries.nnSignificance StatementThe live-attenuated influenza vaccine, in principle provides an important intervention for the control of both seasonal and pandemic influenza. However vaccine effectiveness studies have found seemingly contradictory results with effectiveness ranging from 0 to 50%. Based on mathematical models we suggest that a major factor responsible for the variable efficacy of the vaccine is negative interference - where pre-existing immunity precludes the vaccine from working. Our models suggest that there are broad regimes for which LAIV will fail to be immunogenic, but also allow us to make suggestions for the choice of vaccine strain that will allow optimization of protective immunity in different scenarios.
]]></description>
<dc:creator>Matrajt, L.</dc:creator>
<dc:creator>Halloran, M. E.</dc:creator>
<dc:creator>Antia, R.</dc:creator>
<dc:date>2018-09-28</dc:date>
<dc:identifier>doi:10.1101/424275</dc:identifier>
<dc:title><![CDATA[Successes and failures of the live-attenuated influenza vaccine, can we do better?]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/426288v1?rss=1">
<title>
<![CDATA[
SWI/SNF coordinates transcriptional activation through Rpd3-mediated histone hypoacetylation during quiescence entry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/426288v1?rss=1</link>
<description><![CDATA[
Whether or not a cell chooses to divide is a tightly regulated and extremely important decision. Cells from yeast to human are able to reversibly exit the cell cycle in response to environmental changes such as nutritional changes or removal of growth cues to become quiescent. An inappropriate response to environmental cues can result in overproliferation which can lead to cancer, or a failure to proliferate which can result in developmental defects, premature aging and defects in wound healing. While many of the cell signaling pathways involved in regulating cellular quiescence have been identified, how these pathways translate their messages into transcriptional outputs is not well characterized. We previously showed that the histone deacetylase Rpd3 mediates global histone deacetylation and transcription repression upon quiescence entry. How the activation of quiescence-specific genes occurs in the midst of this transcriptionally repressive environment is not well understood. We show that the SWI/SNF chromatin remodeling complex activates quiescence specific genes to promote entry into quiescence. We additionally show that SWI/SNF binding early during quiescence entry is important for facilitating localization of the transcriptional activator Gis1, as well as histone H4 hypoacetylation in coding regions later on. The increase in H4 acetylation that we observe at Snf2-regulated genes upon Snf2 depletion corresponds to a decrease in promoter-bound Rpd3, suggesting that Snf2 remodels chromatin not only to facilitate activator binding, but also the binding of Rpd3. These observations provide mechanistic insight as to how quiescence-specific genes can be activated in the face of global deacetylation and transcription repression.
]]></description>
<dc:creator>Spain, M. M.</dc:creator>
<dc:creator>Braceros, K. C. A.</dc:creator>
<dc:creator>Tsukiyama, T.</dc:creator>
<dc:date>2018-09-26</dc:date>
<dc:identifier>doi:10.1101/426288</dc:identifier>
<dc:title><![CDATA[SWI/SNF coordinates transcriptional activation through Rpd3-mediated histone hypoacetylation during quiescence entry]]></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/10.1101/431643v1?rss=1">
<title>
<![CDATA[
Differences in protein dosage underlie nongenetic differences in traits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/431643v1?rss=1</link>
<description><![CDATA[
Phenotypic expression of many traits varies among isogenic individuals in homogeneous environments. Intrinsic variation in the protein chaperone system affects a wide variety of traits in diverse biological systems. In C. elegans, expression of hsp-16.2 chaperone biomarkers predicts the penetrance of mutations and lifespan after heat shock. But the physiological mechanisms by which cells express different amounts of the biomarker were unknown. Here, we used an in vivo microscopy approach to dissect the mechanisms of cell-to-cell variation in hsp-16.2 biomarker expression, focusing on the intestines, which generate most signal. We found both intrinsic noise and signaling noise are low. The major axis of cell-to-cell variation in gene expression is composed of general differences in protein dosage. Thus, hsp-16.2 biomarkers reveal states of high or low effective dosages for many genes. It is possible that natural variation in protein dosage or chaperone activity may account for missing heritability of some traits.
]]></description>
<dc:creator>Burnaevskiy, N.</dc:creator>
<dc:creator>Sands, B.</dc:creator>
<dc:creator>Yun, S.</dc:creator>
<dc:creator>Tedesco, P. M.</dc:creator>
<dc:creator>Johnson, T. E.</dc:creator>
<dc:creator>Kaeberlein, M.</dc:creator>
<dc:creator>Brent, R.</dc:creator>
<dc:creator>Mendenhall, A. R.</dc:creator>
<dc:date>2018-10-01</dc:date>
<dc:identifier>doi:10.1101/431643</dc:identifier>
<dc:title><![CDATA[Differences in protein dosage underlie nongenetic differences in traits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/434597v1?rss=1">
<title>
<![CDATA[
Distinct gene-selective roles for a network of core promoter factors in Drosophila neural stem cell identity. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/434597v1?rss=1</link>
<description><![CDATA[
The transcriptional mechanisms that allow neural stem cells (NSC) to balance self-renewal with differentiation are not well understood. Employing an in vivo RNAi screen we identify here NSC-TAFs, a subset of nine TATA-binding protein associated factors (TAFs), as NSC identity genes in Drosophila. We found that depletion of NSC-TAFs results in decreased NSC clone size, reduced proliferation, defective cell polarity and increased hypersensitivity to cell cycle perturbation, without affecting NSC survival. Integrated gene expression and genomic binding analyses revealed that NSC-TAFs function with both TBP and TRF2, and that NSC-TAF-TBP and NSC-TAF-TRF2 shared target genes encode different subsets of transcription factors and RNA-binding proteins with established or emerging roles in NSC identity and brain development. Taken together, our results demonstrate that core promoter factors are selectively required for NSC identity in vivo by promoting cell cycle progression and NSC cell polarity as well as by restraining premature differentiation. Because pathogenic variants in a subset of TAFs have all been linked to human neurological disorders, this work may stimulate and inform future animal models of TAF-linked neurological disorders.nnAuthor summaryThe brains of many animal species are built with brain stem cells. Having too many brain stem cells can lead to brain tumors whereas too few can lead to birth defects such as microcephaly. A number of next generation sequencing studies have implicated proteins referred to as TATA-box-binding protein associated factors (TAFs) in human neurological disorders including microcephaly, but prior to this study, their function in brain development was unknown. Here we use brain stem cells, known as neural stem cells (NSCs), from the fruit fly Drosophila melanogaster as a model system to decipher how TAFs control brain stem cell identity. By combining genetics and low-input genomics, we show that TAFs directly control NSC cell division and cell polarity but do not appear to be required for NSC survival. We further show that TAFs accomplish these functions by associating either with their canonical partner TBP (TATA-binding protein) or the related protein TRF2. In summary, our study reveals unexpected and gene-selective functions of a unique subset of TAFs and their binding partners, which could inform future studies that seek to model human neurological disorders associated with TAFs.
]]></description>
<dc:creator>Neves, A.</dc:creator>
<dc:creator>Eisenman, R. N.</dc:creator>
<dc:date>2018-10-03</dc:date>
<dc:identifier>doi:10.1101/434597</dc:identifier>
<dc:title><![CDATA[Distinct gene-selective roles for a network of core promoter factors in Drosophila neural stem cell identity.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/437277v1?rss=1">
<title>
<![CDATA[
Single-cell virus sequencing of influenza infections that trigger innate immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/437277v1?rss=1</link>
<description><![CDATA[
The outcome of viral infection is extremely heterogeneous, with infected cells only sometimes activating innate immunity. Here we develop a new approach to assess how the genetic variation inherent in viral populations contributes to this heterogeneity. We do this by determining both the transcriptome and full-length sequences of all viral genes in single influenza-infected cells. Most cells are infected by virions with defects such as amino-acid mutations, internal deletions, or failure to express a gene. We identify instances of each type of defect that increase the likelihood that a cell activates an innate-immune response. However, immune activation remains stochastic in cells infected by virions with these defects, and sometimes occurs even when a cell is infected by a virion that expresses unmutated copies of all genes. Our work shows that viral genetic variation substantially contributes to but does not fully explain the heterogeneity in single influenza-infected cells.
]]></description>
<dc:creator>Russell, A. B.</dc:creator>
<dc:creator>Kowalsky, J. R.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2018-10-07</dc:date>
<dc:identifier>doi:10.1101/437277</dc:identifier>
<dc:title><![CDATA[Single-cell virus sequencing of influenza infections that trigger innate immunity]]></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/10.1101/445601v1?rss=1">
<title>
<![CDATA[
Errors in RNA-seq transcript abundance estimates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/445601v1?rss=1</link>
<description><![CDATA[
RNA-sequencing data is widely used to identify disease biomarkers and therapeutic targets. Here, using data from five RNA-seq processing pipelines applied to 6,690 human tumor and normal tissues, we show that for >12% of protein-coding genes, in at least 1% of samples, current best-in-class RNA-seq processing pipelines differ in their abundance estimates by more than four-fold using the same samples and the same set of RNA-seq reads, raising clinical concern.
]]></description>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Pattwell, S. S.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:creator>Bolouri, H.</dc:creator>
<dc:date>2018-10-17</dc:date>
<dc:identifier>doi:10.1101/445601</dc:identifier>
<dc:title><![CDATA[Errors in RNA-seq transcript abundance estimates]]></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/10.1101/446344v1?rss=1">
<title>
<![CDATA[
CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/446344v1?rss=1</link>
<description><![CDATA[
Single cell RNA-seq has emerged as a powerful tool for resolving cellular states associated with normal and maligned developmental processes. Here, we used scRNA-seq to examine the cell cycle states of expanding human neural stem cells (hNSCs). From this data, we created a cell cycle classifier, which, in addition to traditional cell cycle phases, also identifies a putative quiescent-like state in neuroepithelial-derived cell types during mammalian neurogenesis and in gliomas. This state, Neural G0, is enriched for expression of quiescent NSC genes and other neurodevelopmental markers found in non-dividing neural progenitors. For gliomas, Neural G0 cell populations and gene expression is significantly associated with less aggressive tumors and extended patient survival. Genetic screens to identify modulators of Neural G0 revealed that knockout of genes associated with the Hippo/Yap and p53 pathways diminished Neural G0 in vitro, resulting in faster G1 transit, down regulation of quiescence-associated markers, and loss of Neural G0 gene expression. Thus, Neural G0 represents a dynamic quiescent-like state found in neuro-epithelial derived cells and gliomas.
]]></description>
<dc:creator>Feldman, H.</dc:creator>
<dc:creator>Toledo, C.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Hoellerbauer, P.</dc:creator>
<dc:creator>Corrin, P.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Kufeld, M.</dc:creator>
<dc:creator>Bolouri, H.</dc:creator>
<dc:creator>Basom, R.</dc:creator>
<dc:creator>Delrow, J.</dc:creator>
<dc:creator>Meier, J.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>McFaline-Figueroa, J.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Pollard, S.</dc:creator>
<dc:creator>Plaisier, C.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:date>2018-10-17</dc:date>
<dc:identifier>doi:10.1101/446344</dc:identifier>
<dc:title><![CDATA[CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors]]></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/10.1101/453811v1?rss=1">
<title>
<![CDATA[
A Stu2-mediated intrinsic tension-sensing pathway promotes chromosome biorientation in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/453811v1?rss=1</link>
<description><![CDATA[
Accurate segregation of chromosomes to daughter cells is a critical aspect of cell division. It requires the kinetochores on duplicated chromosomes to biorient, attaching to microtubules from opposite poles of the cell. Bioriented attachments come under tension, while incorrect attachments lack tension and must be destabilized. A well-studied error correction pathway is mediated by the Aurora B kinase, which destabilizes low tension-bearing attachments. We recently discovered that in vitro, kinetochores display an additional intrinsic tension-sensing pathway that utilizes Stu2. This pathways contribution to error correction in cells, however, was unknown. Here, we identify a Stu2 mutant that abolishes its kinetochore function and show that it causes error correction defects in vivo. We also show that this intrinsic tension-sensing pathway functions in concert with the Aurora B-mediated pathway. Together, our work indicates that cells employ at least two pathways to ensure biorientation and the accuracy of chromosome segregation.
]]></description>
<dc:creator>Miller, M. P.</dc:creator>
<dc:creator>Evans, R.</dc:creator>
<dc:creator>Zelter, A.</dc:creator>
<dc:creator>Geyer, E.</dc:creator>
<dc:creator>MacCoss, M.</dc:creator>
<dc:creator>Rice, L.</dc:creator>
<dc:creator>Davis, T.</dc:creator>
<dc:creator>Asbury, C.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2018-10-25</dc:date>
<dc:identifier>doi:10.1101/453811</dc:identifier>
<dc:title><![CDATA[A Stu2-mediated intrinsic tension-sensing pathway promotes chromosome biorientation in vivo]]></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/10.1101/454835v1?rss=1">
<title>
<![CDATA[
Connect-seq to superimpose molecular on anatomical neural circuit maps 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/454835v1?rss=1</link>
<description><![CDATA[
The mouse brain contains ~100 million neurons interconnected in a vast array of neural circuits. The identities and functions of individual neuronal components of most circuits are undefined. Here we describe a method, termed  Connect-seq, which combines retrograde viral tracing and single cell transcriptomics to uncover the molecular identities of upstream neurons in a specific circuit and the signaling molecules they use to communicate. Connect-seq can generate a molecular map that can be superimposed on a neuroanatomical map to permit molecular and genetic interrogation of how the neuronal components of a circuit control its function. Application of this method to hypothalamic neurons controlling physiological responses to fear and stress reveal subsets of upstream neurons that express diverse constellations of signaling molecules and can be distinguished by their anatomical locations.
]]></description>
<dc:creator>Hanchate, N. K.</dc:creator>
<dc:creator>Lee, E. J.</dc:creator>
<dc:creator>Ellis, A.</dc:creator>
<dc:creator>Kondoh, K.</dc:creator>
<dc:creator>Kuang, D.</dc:creator>
<dc:creator>Basom, R.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Buck, L. B.</dc:creator>
<dc:date>2018-10-27</dc:date>
<dc:identifier>doi:10.1101/454835</dc:identifier>
<dc:title><![CDATA[Connect-seq to superimpose molecular on anatomical neural circuit maps]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/455972v1?rss=1">
<title>
<![CDATA[
Lineage-dependent differences and the role of IFITM3 in the type-I interferon-induced restriction of Zika virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/455972v1?rss=1</link>
<description><![CDATA[
Type-I interferon (IFN-I) is an important aspect of host innate antiviral response. Recent studies have shown that IFN-I can inhibit Zika virus (ZIKV) replication and that this is mediated in part by Interferon-induced transmembrane protein 3 (IFITM3). ZIKV infections in South America have led to severe congenital syndrome in a subset of infected infants. ZIKV was first identified in Africa, where there is limited evidence for the pathogenic effects associated with the American outbreak, which is fueled by infection with Asian-lineage strains, raising the possibility that the African and Asian ZIKV lineages have distinct pathogenic properties. Given the observation that IFN-I can inhibit ZIKV replication in cell culture, we asked whether ZIKV strains differed in their susceptibility to IFN-I. There was a range of susceptibilities to IFN-I inhibition across virus strains. Virus production in A549 cells was reduced from 3-42-fold for IFN and 63-807-fold for IFN{beta} across a panel of nine viruses, five from the African-lineage and four from the Asian-lineage. African-lineage ZIKV strains were more resistant to IFN-I than Asian-lineage strains, but this difference was only significant for IFN-mediated restriction (p = 0.049). Notably, over-expression of IFITM3 at similar levels induced by IFN-I did not significantly restrict either a prototype African lineage (MR 766) or Asian lineage (PRVABC59) isolate. Moreover, knocking out IFITM3 expression did not result in a significant increase in viral replication or a diminishment of the inhibition by IFN-I. Overall, our findings show that while diverse ZIKV strains are susceptible to the antiviral effects of IFN-I, African-lineage strains are more resistant to IFN. In addition, the majority of the IFN-I-induced inhibition of ZIKV strains cannot be explained by IFITM3, suggesting that other unknown ISGs may be the driving force of the type I IFN response against ZIKV.nnAuthor summaryThe innate immune system, and specifically the type-I interferon response, is a critical component of the host response against viral infections. The recent unprecedented spread and severe pathogenic features of Zika virus in the Americas have led to significant interest in characterizing features of Zika virus strains that have fueled the American outbreak. Zika virus was first identified in Africa, where there is limited evidence for the pathogenic effects associated with the American outbreak. Here, we demonstrate that African-lineage Zika virus strains are significantly more resistant to the effects of type-I interferon, and that type-I interferon-mediated restriction of Zika virus strains is not explained by the host factor Interferon-induced transmembrane protein 3. This improved understanding of Zika virus-host interactions may explain certain pathogenic features of Asian-lineage Zika virus strains that have fueled the American Zika virus epidemic, and supports the search for as-yet-unidentified actors in the interferon response against Zika virus.
]]></description>
<dc:creator>Gobillot, T.</dc:creator>
<dc:creator>Humes, D.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2018-10-29</dc:date>
<dc:identifier>doi:10.1101/455972</dc:identifier>
<dc:title><![CDATA[Lineage-dependent differences and the role of IFITM3 in the type-I interferon-induced restriction of Zika virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/457176v1?rss=1">
<title>
<![CDATA[
High-Risk Human Papillomavirus Oncogenes Disrupt the Fanconi Anemia DNA repair Pathway by Impairing Localization and De-ubiquitination of FancD2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/457176v1?rss=1</link>
<description><![CDATA[
Persistent expression of high-risk HPV oncogenes is necessary for the development of anogenital and oropharyngeal cancers. Here, we show that E6/E7 expressing cells are hypersensitive to DNA crosslinking agent cisplatin and have defects in repairing DNA interstrand crosslinks (ICL). Importantly, we elucidate how E6/E7 attenuate the Fanconi anemia (FA) DNA crosslink repair pathway. Though E6/E7 activated the pathway by increasing FancD2 monoubiquitination and foci formation, they inhibited the completion of the repair by multiple mechanisms. E6/E7 impaired FancD2 colocalization with double-strand breaks (DSB), which subsequently hindered the recruitment of downstream protein Rad51 to DSB in E6 cells. Further, E6 expression caused delayed FancD2 de-ubiquitination, an important process for effective ICL repair. Delayed FancD2 de-ubiquitination was attributed to the increased chromatin retention of FancD2 hindering USP1 de-ubiquitinating activity, and persistently activated ATR/CHK-1/pS565 FancI signaling. E6 mediated p53 degradation did not hamper the cell cycle specific process of FancD2 modifications but abrogated repair by disrupting FancD2 de-ubiquitination. Further, E6 reduced the expression and foci formation of Palb2, which is a repair protein downstream of FancD2. These findings uncover unique mechanisms by which HPV oncogenes contribute to genomic instability and the response to cisplatin therapies.nnAUTHOR SUMMARYHigh-risk human papillomavirus (HPV) causes nearly all cervical and many other anogenital cancers, and oropharyngeal cancers. As cisplatin is the most commonly used drug for cervical and HPV-associated oropharyngeal cancers, it is important to understand how HPV oncogenes disrupt the Fanconi anemia (FA) pathway involved primarily in the repair of cisplatin-induced DNA crosslinks. However, the mechanism by which HPV E6 and E7 attenuate the FA pathway is poorly understood. We demonstrate that E6/E7 expression disrupts crosslink repair and increase cisplatin sensitivity, and attenuate the FA pathway through multiple unique mechanisms. First, E6/E7 causes accumulation of FancD2, a central component of the FA pathway, at the sites away from DNA damage. This results in reduced recruitment of Rad51, another repair protein involved in the pathway. Second, E6 causes delayed FancD2 de-ubiquitination, an important process for effective repair. Third, E6 expressing cells decreases the expression and foci formation of Palb2 repair protein. Together, this work elucidates the mechanisms by which HPV attenuates the repair of DNA crosslinks increasing cisplatin cytotoxicity and efficacy in treating HPV-positive cancers.
]]></description>
<dc:creator>Khanal, S.</dc:creator>
<dc:creator>Galloway, D. A.</dc:creator>
<dc:date>2018-10-30</dc:date>
<dc:identifier>doi:10.1101/457176</dc:identifier>
<dc:title><![CDATA[High-Risk Human Papillomavirus Oncogenes Disrupt the Fanconi Anemia DNA repair Pathway by Impairing Localization and De-ubiquitination of FancD2]]></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/10.1101/457648v1?rss=1">
<title>
<![CDATA[
N6-methyladenosine mRNA marking promotes selective translation of regulons required for human erythropoiesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/457648v1?rss=1</link>
<description><![CDATA[
Many of the regulatory features governing erythrocyte specification, maturation, and associated disorders remain enigmatic. To identify new regulators of erythropoiesis, we performed a functional genomic screen for genes affecting expression of the erythroid marker CD235a/GYPA. Among validating hits were genes coding for the N6-methyladenosine (m6A) mRNA methyltransferase (MTase) complex, including, METTL14, METTL3, and WTAP. We found that m6A MTase activity promotes erythroid gene expression programs and lineage specification through selective translation of >200 m6A marked mRNAs, including those coding for SETD methyltransferase, ribosome, and polyA RNA binding proteins. Remarkably, loss of m6A marks resulted in dramatic loss of H3K4me3 across key erythroid-specific KLF1 transcriptional targets (e.g., Heme biosynthesis genes). Further, each m6A MTase subunit and a subset of their mRNAs targets, including BRD7, CXXC1, PABPC1, PABPC4, STK40, and TADA2B, were required for erythroid specification. Thus, m6A mRNA marks promote the translation of a network of genes required for human erythropoiesis.
]]></description>
<dc:creator>Kuppers, D.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Lim, Y.</dc:creator>
<dc:creator>Lim, A.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Corrin, P.</dc:creator>
<dc:creator>Plasier, C.</dc:creator>
<dc:creator>Basom, R.</dc:creator>
<dc:creator>Delrow, J.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>He, H.</dc:creator>
<dc:creator>Torok-Storb, B.</dc:creator>
<dc:creator>Hsieh, A.</dc:creator>
<dc:creator>Paddison, P.</dc:creator>
<dc:date>2018-10-31</dc:date>
<dc:identifier>doi:10.1101/457648</dc:identifier>
<dc:title><![CDATA[N6-methyladenosine mRNA marking promotes selective translation of regulons required for human erythropoiesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/461004v1?rss=1">
<title>
<![CDATA[
Killer meiotic drive and dynamic evolution of the wtf gene family 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/461004v1?rss=1</link>
<description><![CDATA[
Natural selection works best when the two alleles in a diploid organism are transmitted to offspring at equal frequencies. Despite this, selfish loci known as meiotic drivers that bias their own transmission into gametes are found throughout eukaryotes. Drive is thought to be a powerful evolutionary force, but empirical evolutionary analyses of drive systems are limited by low numbers of identified meiotic drive genes. Here, we analyze the evolution of the wtf gene family of Schizosaccharomyces pombe that contains both killer meiotic drive genes and suppressors of drive. We completed assemblies of all wtf genes for two S. pombe strains, as well as a subset of wtf genes from over 50 strains. We find that wtf copy number can vary greatly between strains, and that amino acid substitutions, expansions and contractions of DNA sequence repeats, and nonallelic gene conversion between family members all contribute to dynamic wtf gene evolution. This work demonstrates the power of meiotic drive to foster rapid evolution and identifies a recombination mechanism through which transposons can indirectly mobilize meiotic drivers.
]]></description>
<dc:creator>Eickbush, M. T.</dc:creator>
<dc:creator>Young, J. M.</dc:creator>
<dc:creator>Zanders, S. E.</dc:creator>
<dc:date>2018-11-03</dc:date>
<dc:identifier>doi:10.1101/461004</dc:identifier>
<dc:title><![CDATA[Killer meiotic drive and dynamic evolution of the wtf gene family]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/471094v1?rss=1">
<title>
<![CDATA[
Transgenic mice expressing tunable levels of DUX4 develop characteristic facioscapulohumeral muscular dystrophy-like pathophysiology ranging in severity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/471094v1?rss=1</link>
<description><![CDATA[
BackgroundAll types of facioscapulohumeral muscular dystrophy (FSHD) are caused by the aberrant myogenic activation of the somatically silent DUX4 gene, which initiates a cascade of cellular events ultimately leading to FSHD pathophysiology. Therefore, FSHD is a dominant gain-of-function disease that is amenable to modeling by DUX4 overexpression. However, there is large variability in the patient population. Typically, progressive skeletal muscle weakness becomes noticeable in the second or third decade of life, yet there are many genetically FSHD individuals who develop symptoms much later in life or remain relatively asymptomatic throughout their lives. Conversely, in rare cases, FSHD may present clinically prior to 5-10 yrs of age, ultimately manifesting as a very severe early onset form of the disease. Thus, there is a need to control the timing and severity of pathology in FSHD-like models.nnMethodsWe have recently described a line of conditional DUX4 transgenic mice, FLExDUX4, that develop a myopathy upon induction of human DUX4-fl expression in skeletal muscle. Here, we use the FLExDUX4 mouse crossed with the skeletal muscle-specific and tamoxifen inducible line ACTAl-MerCreMer to generate a highly versatile bi-transgenic mouse model with chronic, low-level DUX4-fl expression and mild pathology, that can be induced to develop more severe FSHD-like pathology in a dose-dependent response to tamoxifen. We identified conditions to reproducibly generate models exhibiting mild, moderate, or severe DUX4-dependent pathophysiology, and characterized their progression.nnResultsWe assayed DUX4-fl mRNA and protein levels, fitness, strength, global gene expression, histopathology, and immune response, all of which are consistent with an FSHD-like myopathic phenotype. Importantly, we identified sex-specific and muscle-specific differences that should be considered when using these models for preclinical studies.nnConclusionsThe ACTA1-MCM;FLExDUX4 bi-transgenic mouse model expresses a chronic low level of DUX4-fl and has mild pathology and detectable muscle weakness. The onset and progression of moderate to severe pathology can be controlled via tamoxifen injection to provide consistent and readily screenable phenotypes for assessing therapies targeting DUX4-fl mRNA and protein. Thus, these FSHD-like mouse models can be used to study a range of DUX4-fl expression and pathology dependent upon investigator need, through controlled mosaic expression of DUX4.
]]></description>
<dc:creator>Jones, T. I.</dc:creator>
<dc:creator>Chew, G.-L.</dc:creator>
<dc:creator>Barraza-Flores, P.</dc:creator>
<dc:creator>Schreier, S.</dc:creator>
<dc:creator>Ramirez, M.</dc:creator>
<dc:creator>Wuebbles, R. D.</dc:creator>
<dc:creator>Burkin, D. J.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:creator>Jones, P. L.</dc:creator>
<dc:date>2018-11-15</dc:date>
<dc:identifier>doi:10.1101/471094</dc:identifier>
<dc:title><![CDATA[Transgenic mice expressing tunable levels of DUX4 develop characteristic facioscapulohumeral muscular dystrophy-like pathophysiology ranging in severity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/472746v1?rss=1">
<title>
<![CDATA[
Massively parallel profiling of HIV-1 resistance to the fusion inhibitor enfuvirtide 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/472746v1?rss=1</link>
<description><![CDATA[
Resistance to enfuvirtide, the only clinically approved HIV-1 entry inhibitor, has primarily been mapped to the binding site in the N-terminal heptad repeat (NHR) of the Env transmembrane domain and a limited number of allosteric sites. To better delineate the genotypic determinants of resistance, we used deep mutational scanning to quantify how all mutations to HIV-1 Env affect enfuvirtide sensitivity. We identified numerous additional resistance mutations in the NHR and other regions of Env, including the co-receptor binding site. This complete map of resistance sheds light on the diverse mechanisms of enfuvirtide resistance and can inform clinical monitoring of patients.
]]></description>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Arenz, D.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2018-11-19</dc:date>
<dc:identifier>doi:10.1101/472746</dc:identifier>
<dc:title><![CDATA[Massively parallel profiling of HIV-1 resistance to the fusion inhibitor enfuvirtide]]></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/10.1101/488478v1?rss=1">
<title>
<![CDATA[
Separate Polycomb Response Elements control chromatin state and activation of the vestigial gene 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/488478v1?rss=1</link>
<description><![CDATA[
Patterned expression of many developmental genes is specified by transcription factor gene expression, but is thought to be refined by chromatin-mediated repression. Regulatory DNA sequences called Polycomb Response Elements (PREs) are required to repress some developmental target genes, and are widespread in genomes, suggesting that they broadly affect developmental programs. While PREs in transgenes can nucleate trimethylation on lysine 27 of the histone H3 tail (H3K27me3), none have been demonstrated to be necessary at endogenous chromatin domains. This failure is thought to be due to the fact that most endogenous H3K27me3 domains contain many PREs, and individual PREs may be redundant. In contrast to these ideas, we show here that PREs near the wing selector gene vestigial have distinctive roles at their endogenous locus, even though both PREs are repressors in transgenes. First, a PRE near the promoter is required for vestigial activation and not for repression. Second, only the distal PRE contributes to H3K27me3, but even removal of both PREs does not eliminate H3K27me3 across the vestigial domain. Thus, endogenous chromatin domains appear to be intrinsically marked by H3K27me3, and PREs appear required to enhance this chromatin modification to high levels at inactive genes.nnAuthor summaryEukaryotic genes are packaged in chromatin, and their transcription relies on activators that recruit RNA polymerases and on repressive factors. Some of these factors factors that modulate chromatin structure to promote or inhibit gene transcription. In multicellular organisms, cell types have distinct patterns of gene expression, and these patterns are controlled by by the expression of cell-type-specific transcription factors and by modulating chromatin structure. The Polycomb system is one major system for the chromatin-mediated silencing of developmental gene expression, where a histone methyltransferase marks extended chromatin domains with trimethylation of lysine-27 of the histone H3 tail (H3K27me3) and forms repressed chromatin. In Drosophila, repressive regulatory elements called Polycomb Response Elements (PREs) are thought to nucleate histone methyltransferase binding which then spreads across these domains. In this study, we demonstrate that two PREs near the developmental vestigial gene have distinct and separable effects on gene activation and chromatin structure. Both PREs are functional repressors in transgenes, but the PRE located near the vestigial promoter is required for gene transcription. This PRE has no effect on histone methylation of the domain. The second PRE, located in the middle of the chromatin domain is required for high-level H3K27me3 of the domain, and this methylation is not required to refine vestigial gene expression. Strikingly, a significant chromatin methylation remains when both PREs are deleted. Our findings imply that PREs near promoters may often play activating roles in gene expression in the Drosophila genome. We suggest that some domains of H3K27me3 occur in regions that lack histone acetylation, and has little consequence for correctly patterning gene expression.
]]></description>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Spens, A. E.</dc:creator>
<dc:date>2018-12-05</dc:date>
<dc:identifier>doi:10.1101/488478</dc:identifier>
<dc:title><![CDATA[Separate Polycomb Response Elements control chromatin state and activation of the vestigial gene]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/512525v1?rss=1">
<title>
<![CDATA[
Comprehensive mapping of avian influenza polymerase adaptation to the human host 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/512525v1?rss=1</link>
<description><![CDATA[
Viruses like influenza are infamous for their ability to adapt to new hosts. Retrospective studies of natural zoonoses and passaging in the lab have identified a modest number of host-adaptive mutations. However, it is unclear if these mutations represent all ways that influenza can adapt to a new host. Here we take a prospective approach to this question by completely mapping amino-acid mutations to the avian influenza virus polymerase protein PB2 that enhance growth in human cells. We identify numerous previously uncharacterized human-adaptive mutations. These mutations cluster on PB2s surface, highlighting potential interfaces with host factors. Some previously uncharacterized adaptive mutations occur in avian-to-human transmission of H7N9 influenza, showing their importance for natural virus evolution. But other adaptive mutations do not occur in nature because they are inaccessible via single-nucleotide mutations. Overall, our work shows how selection at key molecular surfaces combines with evolutionary accessibility to shape viral host adaptation.
]]></description>
<dc:creator>Soh, Y. Q. S.</dc:creator>
<dc:creator>Moncla, L. H.</dc:creator>
<dc:creator>Eguia, R.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2019-01-04</dc:date>
<dc:identifier>doi:10.1101/512525</dc:identifier>
<dc:title><![CDATA[Comprehensive mapping of avian influenza polymerase adaptation to the human host]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/512749v1?rss=1">
<title>
<![CDATA[
RNAIndel: a machine-learning framework for discovery of somatic coding indels using tumor RNA-Seq data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/512749v1?rss=1</link>
<description><![CDATA[
Reliable identification of expressed somatic insertion/deletion (indels) is an unmet demand due to artifacts generated in PCR-based RNA-Seq library preparation and the lack of normal RNA-Seq data, presenting analytical challenges for discovery of somatic indels in tumor trasncriptome.nnBy implementing features characterized by PCR-free whole-genome and whole-exome sequencing into a machine-learning framework, we present RNAIndel, a tool for predicting somatic, germline and artifact indels from tumor RNA-Seq data alone. RNAIndel robustly predicts 87{square}93% of somatic indels from 235 samples with heterogeneous conditions, even recovering subclonal (VAF range 0.01-0.15) driver indels missed by targeted deep-sequencing, outperforming the current best-practice for RNA-Seq variant calling which had 57% sensitivity but with 12 times more false positives.nnRNAIndel is freely available at https://github.com/stjude/RNAIndelnnContactjinghui.zhang@stjude.org
]]></description>
<dc:creator>Hagiwara, K.</dc:creator>
<dc:creator>Ding, L.</dc:creator>
<dc:creator>Edmonson, M. N.</dc:creator>
<dc:creator>Rice, S. V.</dc:creator>
<dc:creator>Newman, S.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Ries, R. E.</dc:creator>
<dc:creator>Rusch, M.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:date>2019-01-07</dc:date>
<dc:identifier>doi:10.1101/512749</dc:identifier>
<dc:title><![CDATA[RNAIndel: a machine-learning framework for discovery of somatic coding indels using tumor RNA-Seq data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/526475v1?rss=1">
<title>
<![CDATA[
Fate plasticity and reprogramming in genetically distinct populations of Danio leucophores 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/526475v1?rss=1</link>
<description><![CDATA[
Understanding genetic and cellular bases of adult form remains a fundamental goal at the intersection of developmental and evolutionary biology. The skin pigment cells of vertebrates, derived from embryonic neural crest, are a useful system for elucidating mechanisms of fate specification, pattern formation, and how particular phenotypes impact organismal behavior and ecology. In a survey of Danio fishes, including zebrafish Danio rerio, we identified two populations of white pigment cells--leucophores--one of which arises by transdifferentiation of adult melanophores and another that develops from a yellow/orange xanthophore-like progenitor. Single-cell transcriptomic, mutational, chemical and ultrastructural analyses of zebrafish leucophores revealed cell-type specific chemical compositions, organelle configurations and genetic requirements. At the organismal level, we identified distinct physiological responses of leucophores during environmental background matching and we show that leucophore complement influences behavior. Together, our studies revealed new, independently arisen pigment cell types and mechanisms of fate acquisition in zebrafish, and illustrate how concerted analyses across hierarchical levels can provide insights into phenotypes and their evolution.
]]></description>
<dc:creator>Lewis, V.</dc:creator>
<dc:creator>Saunders, L.</dc:creator>
<dc:creator>Larson, T. A.</dc:creator>
<dc:creator>Bain, E.</dc:creator>
<dc:creator>Sturiale, S.</dc:creator>
<dc:creator>Gur, D.</dc:creator>
<dc:creator>Chowdhury, S.</dc:creator>
<dc:creator>Flynn, J.</dc:creator>
<dc:creator>Allen, M.</dc:creator>
<dc:creator>Deheyn, D.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Simon, J.</dc:creator>
<dc:creator>Lippincott-Schwartz, J.</dc:creator>
<dc:creator>Raible, D.</dc:creator>
<dc:creator>Parichy, D.</dc:creator>
<dc:date>2019-01-22</dc:date>
<dc:identifier>doi:10.1101/526475</dc:identifier>
<dc:title><![CDATA[Fate plasticity and reprogramming in genetically distinct populations of Danio leucophores]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/530832v1?rss=1">
<title>
<![CDATA[
Antigen-specific humoral immune responses by CRISPR/Cas9-edited B cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/530832v1?rss=1</link>
<description><![CDATA[
A small number of HIV-1 infected individuals develop broadly neutralizing-antibodies to the virus (bNAbs). These antibodies are protective against infection in animal models. However, they only emerge 1 - 3 years after infection, and show a number of highly unusual features including exceedingly high levels of somatic mutations. It is therefore not surprising that elicitation of protective immunity to HIV-1 has not yet been possible. Here we show that mature, primary mouse and human B cells can be edited in vitro using CRISPR/Cas9 to express mature bNAbs from the endogenous Igh locus. Moreover, edited B cells retain the ability to participate in humoral immune responses. Immunization with cognate antigen in wild type mouse recipients of edited B cells elicits bNAb titers that neutralize HIV-1 at levels associated with protection against infection. This approach enables humoral immune responses that may be difficult to elicit by traditional immunization.

One-sentence summaryB cells edited by CRISPR/Cas9 to produce antibodies participate in humoral immune reactions and secrete neutralizing serum titers of anti-HIV bNAbs.
]]></description>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Horning, M.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:creator>Dosenovic, P.</dc:creator>
<dc:creator>Yost, D.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2019-01-26</dc:date>
<dc:identifier>doi:10.1101/530832</dc:identifier>
<dc:title><![CDATA[Antigen-specific humoral immune responses by CRISPR/Cas9-edited B cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/531236v1?rss=1">
<title>
<![CDATA[
Tumor-Intrinsic Response to IFNgamma Shapes the Tumor Microenvironment and Anti-PD-1 Response in NSCLC 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/531236v1?rss=1</link>
<description><![CDATA[
Targeting PD-1/ PD-L1 is only effective in ~20% of lung cancer patients, but determinants of this response are poorly defined. We previously observed differential responses of two murine K-Ras lung cancer cell lines to anti-PD-1 therapy: CMT167 tumors were eliminated while LLC tumors were resistant. The goal of this study was to define mechanism(s) mediating this difference. RNA-Seq analysis of cancer cells recovered from lung tumors revealed that CMT167 cells induced an IFN{gamma} signature that was absent in LLC cells. Silencing Ifngr1 in CMT167 resulted in tumors resistant to IFN{gamma} and anti-PD-1 therapy. Conversely, LLC cells had high basal expression of Socs1, an inhibitor of IFN{gamma}. Silencing Socs1 increased response to IFN{gamma} in vitro and sensitized tumors to anti-PD-1. This was associated with a reshaped TME, characterized by enhanced T cell infiltration and enrichment of PD-L1 high myeloid cells. These studies demonstrate that targeted enhancement of tumor-intrinsic IFN{gamma} signaling can induce of cascade of changes associated with increased therapeutic vulnerability.

SummaryMechanisms regulating response to anti-PD-1 therapy in lung cancer are not well defined. This study, using orthotopic immunocompetent mouse models of lung cancer, demonstrates that intrinsic sensitivity of cancer cells to IFN{gamma} determines anti-PD-1 responsiveness through alterations in the tumor microenvironment.
]]></description>
<dc:creator>Bullock, B. L.</dc:creator>
<dc:creator>Kimball, A. K.</dc:creator>
<dc:creator>Poczobutt, J. M.</dc:creator>
<dc:creator>Li, H. Y.</dc:creator>
<dc:creator>Kwak, J. W.</dc:creator>
<dc:creator>Neuwelt, A. J.</dc:creator>
<dc:creator>Johnson, A. M.</dc:creator>
<dc:creator>Kleczko, E.</dc:creator>
<dc:creator>Kaspar, R.</dc:creator>
<dc:creator>Hopp, K.</dc:creator>
<dc:creator>Schenk, E.</dc:creator>
<dc:creator>Weiser-Evans, M. C.</dc:creator>
<dc:creator>Clambey, E. T.</dc:creator>
<dc:creator>Nemenoff, R. A.</dc:creator>
<dc:date>2019-01-27</dc:date>
<dc:identifier>doi:10.1101/531236</dc:identifier>
<dc:title><![CDATA[Tumor-Intrinsic Response to IFNgamma Shapes the Tumor Microenvironment and Anti-PD-1 Response in NSCLC]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/541979v1?rss=1">
<title>
<![CDATA[
B cells engineered to express pathogen-specific antibodies using CRISPR/Cas9 protect against infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/541979v1?rss=1</link>
<description><![CDATA[
Effective vaccines inducing lifelong protection against many important infections such as respiratory syncytial virus (RSV), human immunodeficiency virus (HIV), influenza and Epstein-Barr virus (EBV) are not yet available despite decades of research. As an alternative to a protective vaccine we developed a genetic engineering strategy in which CRISPR/Cas9 was utilized to replace endogenously-encoded antibodies with antibodies protective against RSV, HIV, influenza or EBV in primary human or murine B cells. The engineered antibodies were expressed in up to 59% of primary B cells under the control of endogenous regulatory elements, which maintained normal antibody expression and secretion. Importantly, a single transfer of murine B cells engineered to express an antibody protective against RSV resulted in potent and durable protection against RSV infection in immunocompromised hosts. This approach offers the opportunity to achieve sterilizing immunity against pathogens for which traditional vaccination has failed to induce or maintain protective antibody responses.
]]></description>
<dc:creator>Moffett, H. F.</dc:creator>
<dc:creator>Harms, C. K.</dc:creator>
<dc:creator>Fitzpatrick, K. S.</dc:creator>
<dc:creator>Tooley, M. R.</dc:creator>
<dc:creator>Boonyaratankornkit, J.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:date>2019-02-06</dc:date>
<dc:identifier>doi:10.1101/541979</dc:identifier>
<dc:title><![CDATA[B cells engineered to express pathogen-specific antibodies using CRISPR/Cas9 protect against infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/543132v1?rss=1">
<title>
<![CDATA[
Proteomic and Transcriptomic Profiling Identifies Mediators of Anchorage-Independent Growth and Roles of Inhibitor of Differentiation Proteins in Invasive Lobular Breast Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/543132v1?rss=1</link>
<description><![CDATA[
BACKGROUNDInvasive lobular carcinoma (ILC) is a histological subtype of breast cancer with distinct molecular and clinical features from the more common subtype invasive lobular carcinoma (IDC). We have previously shown that human ILC cells lines have a remarkably unique ability to grow in ultra-low attachment (ULA) suspension cultures as compared to IDC cells, the mediators of which remain unknown.

METHODSUsing flow cytometry and immunoblotting in human ILC and IDC cell lines, we measured levels of apoptosis and cell proliferation in attached (2D) and suspension (ULA) cultures. siRNA-mediated knockdown and pharmacological inhibitors were utilized to assess the effects of known regulators of anchorage-independence. Reverse Phase Protein Arrays and RNA-Sequencing were performed to identify novel proteomic and transcriptomic mediators of ULA growth in ILC cells.

RESULTSWe show that human ILC cell lines exhibit enhanced anoikis resistance and cell proliferation in ULA cultures as compared to IDC cells. Transient restoration of E-cadherin did not impact the 2D or ULA growth of human ILC cell lines, while transient E-cadherin knockdown in IDC cells partially rescued their growth defect in ULA culture. Inhibition of the Rho/ROCK, p120-catenin or YAP/Hippo pathways previously implicated in anoikis resistance did not have a major effect on the ULA growth of ILC cells. Proteomic comparison of ILC and IDC cell lines identified unique induction of PI3K/Akt and p90-RSK pathways in ULA culture in ILC cells. Transcriptional profiling uncovered unique upregulation of the Inhibitors of Differentiation family transcription factors ID1 and ID3 in ILC ULA culture, the knockdown of which diminished anchorage-independent growth. We find that ID1 and ID3 expression is higher in human ILC tumors as compared to IDC and correlated with a worse disease-specific survival uniquely in the ILC cohort.

CONCLUSIONOur comprehensive study of 2D and ULA growth in human ILC cell lines revealed anoikis resistance, cell proliferation and novel mediators of anchorage-independence and provides possible mechanistic insights and clinical implications for metastatic dissemination of ILC. High expression in human ILC tumors and association with clinical outcome implicate ID1 and ID3 as novel drivers and therapeutic targets for lobular breast cancer.
]]></description>
<dc:creator>Tasdemir, N.</dc:creator>
<dc:creator>Ding, K.</dc:creator>
<dc:creator>Levine, K. M.</dc:creator>
<dc:creator>Du, T.</dc:creator>
<dc:creator>Bossart, E. A.</dc:creator>
<dc:creator>Lee, A. V.</dc:creator>
<dc:creator>Davidson, N. E.</dc:creator>
<dc:creator>Oesterreich, S.</dc:creator>
<dc:date>2019-02-07</dc:date>
<dc:identifier>doi:10.1101/543132</dc:identifier>
<dc:title><![CDATA[Proteomic and Transcriptomic Profiling Identifies Mediators of Anchorage-Independent Growth and Roles of Inhibitor of Differentiation Proteins in Invasive Lobular Breast Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/556522v1?rss=1">
<title>
<![CDATA[
Max deletion destabilizes MYC protein and abrogates Eμ-Myc lymphomagenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/556522v1?rss=1</link>
<description><![CDATA[
Although MAX is widely regarded as an obligate dimerization partner for MYC, its function in normal development and neoplasia is not well defined. We show that B-cell specific deletion of Max has a surprisingly modest effect on B-cell development but completely abrogates E{micro}-Myc driven lymphomagenesis. In both contexts, MAX loss leads to a significant reduction in MYC protein levels. This outcome is associated with the downregulation of numerous transcriptional targets of MAX including a subset that regulate MYC stability. Reduction in MYC protein levels is also observed in multiple cell lines treated with a MYC-MAX dimerization inhibitor. Our work uncovers a layer of Myc autoregulation critical for lymphomagenesis yet partly dispensable for normal lymphoid development.
]]></description>
<dc:creator>Mathsyaraja, H.</dc:creator>
<dc:creator>Freie, B.</dc:creator>
<dc:creator>Cheng, P.-F.</dc:creator>
<dc:creator>Babaeva, E.</dc:creator>
<dc:creator>Janssens, D.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Eisenman, R. N.</dc:creator>
<dc:date>2019-02-21</dc:date>
<dc:identifier>doi:10.1101/556522</dc:identifier>
<dc:title><![CDATA[Max deletion destabilizes MYC protein and abrogates Eμ-Myc lymphomagenesis]]></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/10.1101/557264v1?rss=1">
<title>
<![CDATA[
Evolution-guided design of super-restrictor antiviral proteins reveals a breadth-versus-specificity tradeoff 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/557264v1?rss=1</link>
<description><![CDATA[
Antagonistic interactions drive host-virus evolutionary arms-races, which often manifest as recurrent amino acid changes (i.e., positive selection) at their protein-protein interaction interfaces. Here, we investigated whether combinatorial mutagenesis of positions under positive selection in a host antiviral protein could enhance its restrictive properties. We tested ~700 variants of human MxA, generated by combinatorial mutagenesis, for their ability to restrict Thogoto orthomyxovirus (THOV). We identified MxA super-restrictors with increased binding to THOV NP target protein and 10-fold higher anti-THOV restriction relative to wild-type human MxA, the most potent naturally-occurring anti-THOV restrictor identified. Our findings reveal a means to elicit super-restrictor antiviral proteins by leveraging signatures of positive selection. Although some MxA super-restrictors of THOV were impaired in their restriction of H5N1 influenza A virus (IAV), other super-restrictor variants increased THOV restriction without impairment of IAV restriction. Thus, broadly acting antiviral proteins such as MxA mitigate breadth-versus-specificity tradeoffs that could otherwise constrain their adaptive landscape.
]]></description>
<dc:creator>Colon-Thillet, R. S.</dc:creator>
<dc:creator>Hsieh, E. S.</dc:creator>
<dc:creator>Graf, L.</dc:creator>
<dc:creator>McLaughlin, R.</dc:creator>
<dc:creator>Kochs, G.</dc:creator>
<dc:creator>Emerman, M. S.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2019-02-22</dc:date>
<dc:identifier>doi:10.1101/557264</dc:identifier>
<dc:title><![CDATA[Evolution-guided design of super-restrictor antiviral proteins reveals a breadth-versus-specificity tradeoff]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/561936v1?rss=1">
<title>
<![CDATA[
The kinesin-like protein Pavarotti functions non-canonically to regulate actin dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/561936v1?rss=1</link>
<description><![CDATA[
Pavarotti, the Drosophila MKLP1 ortholog, is a kinesin-like protein that with Tumbleweed (MgcRacGAP) works together as the centralspindlin complex. This complex is essential for cytokinesis where it helps to organize the contractile actomyosin ring at the equator of dividing cells by activating the RhoGEF Pebble. Actomyosin rings also function as the driving force during cell wound repair. We previously showed that Tumbleweed and Pebble are required for the cell wound repair process. Here, we show that Pavarotti also functions during wound repair and confirm that while Pavarotti, Tumbleweed, and Pebble are utilized during this cellular repair, it is not as the conserved centralspindlin complex. Surprisingly, in vitro and in vivo work show that the classically microtubule-associated Pavarotti binds directly to actin and has a non-canonical role directly regulating actin dynamics. We show that Pavarotti also works independently from Tumbleweed in several actin-related processes during the normal developmental process of oogenesis.
]]></description>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Verboon, J. M.</dc:creator>
<dc:creator>Prentiss, C. L.</dc:creator>
<dc:creator>Parkhurst, S. M.</dc:creator>
<dc:date>2019-02-26</dc:date>
<dc:identifier>doi:10.1101/561936</dc:identifier>
<dc:title><![CDATA[The kinesin-like protein Pavarotti functions non-canonically to regulate actin dynamics]]></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/10.1101/562371v1?rss=1">
<title>
<![CDATA[
Inverted translational control of eukaryotic gene expression by ribosome collisions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/562371v1?rss=1</link>
<description><![CDATA[
The canonical model of eukaryotic translation posits that efficient translation initiation increases protein expression and mRNA stability. Contrary to this dogma, we show that increasing initiation rate can decrease both protein expression and stability of certain mRNAs in the budding yeast, S. cerevisiae. These mRNAs contain a stretch of poly-basic residues that cause ribosome stalling. Using computational modeling, we predict that the observed decrease in gene expression at high initiation rates occurs when ribosome collisions at stalls stimulate abortive termination of the leading ribosome and cause endonucleolytic mRNA cleavage. We test our prediction by identifying critical roles for the collision-associated quality control factors, Asc1 and Hel2 (RACK1 and ZNF598 in humans, respectively). Remarkably, hundreds of S. cerevisiae mRNAs that contain ribosome-stall sequences also exhibit lower translation efficiency. We propose that these mRNAs have undergone evolutionary selection for inefficient initiation to escape collision-stimulated reduction in gene expression.
]]></description>
<dc:creator>Park, H.</dc:creator>
<dc:creator>Subramaniam, A. R.</dc:creator>
<dc:date>2019-02-27</dc:date>
<dc:identifier>doi:10.1101/562371</dc:identifier>
<dc:title><![CDATA[Inverted translational control of eukaryotic gene expression by ribosome collisions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/567818v1?rss=1">
<title>
<![CDATA[
Clustering co-abundant genes identifies components of the gut microbiome that are reproducibly associated with colorectal cancer and inflammatory bowel disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/567818v1?rss=1</link>
<description><![CDATA[
BackgroundWhole-genome "shotgun" (WGS) metagenomic sequencing is an increasingly widely used tool for analyzing the metagenomic content of microbiome samples. While WGS data contains gene-level information, it can be challenging to analyze the millions of microbial genes which are typically found in microbiome experiments. To mitigate the ultrahigh dimensionality challenge of gene-level metagenomics, it has been proposed to cluster genes by co-abundance to form Co-Abundant Gene groups (CAGs). However, exhaustive co-abundance clustering of millions of microbial genes across thousands of biological samples has previously been intractable purely due to the computational challenge of performing trillions of pairwise comparisons.nnResultsHere we present a novel computational approach to the analysis of WGS datasets in which microbial gene groups are the fundamental unit of analysis. We use the Approximate Nearest Neighbor heuristic for near-exhaustive average linkage clustering to group millions of genes by co-abundance. This results in thousands of high-quality CAGs representing complete and partial microbial genomes. We applied this method to publicly available WGS microbiome surveys and found that the resulting microbial CAGs associated with inflammatory bowel disease (IBD) and colorectal cancer (CRC) were highly reproducible and could be validated independently using multiple independent cohorts.nnConclusionsThis powerful approach to gene-level metagenomics provides a powerful path forward for identifying the biological links between the microbiome and human health. By proposing a new computational approach for handling high dimensional metagenomics data, we identified specific microbial gene groups that are associated with disease that can be used to identify strains of interest for further preclinical and mechanistic experimentation.
]]></description>
<dc:creator>Minot, S. S.</dc:creator>
<dc:creator>Willis, A. D.</dc:creator>
<dc:date>2019-03-05</dc:date>
<dc:identifier>doi:10.1101/567818</dc:identifier>
<dc:title><![CDATA[Clustering co-abundant genes identifies components of the gut microbiome that are reproducibly associated with colorectal cancer and inflammatory bowel disease]]></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/10.1101/569095v1?rss=1">
<title>
<![CDATA[
Functional metagenomics-guided discovery of potent Cas9 inhibitors in the human microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/569095v1?rss=1</link>
<description><![CDATA[
CRISPR-Cas systems protect bacteria and archaea from phages and other mobile genetic elements, which use small anti-CRISPR (Acr) proteins to overcome CRISPR-Cas immunity. Because they are difficult to identify, the natural diversity and impact of Acrs on microbial ecosystems is underappreciated. To overcome this discovery bottleneck, we developed a high-throughput functional selection that isolates acr genes based on their ability to inhibit CRISPR-Cas function. Using this selection, we discovered ten DNA fragments from human oral and fecal metagenomes that antagonize Streptococcus pyogenes Cas9 (SpyCas9). The most potent acr discovered, acrIIA11, was recovered from a Lachnospiraceae phage and is among the strongest known SpyCas9 inhibitors. AcrIIA11 homologs are distributed across multiple bacterial phyla and many divergent homologs inhibit SpyCas9. We show that AcrIIA11 antagonizes SpyCas9 using a different mechanism than that of previously characterized inhibitors. Our study highlights the power of functional selections to uncover widespread Cas9 inhibitors within diverse microbiomes.
]]></description>
<dc:creator>Forsberg, K. J.</dc:creator>
<dc:creator>Bhatt, I. V.</dc:creator>
<dc:creator>Schmidtke, D. T.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>Kaiser, B. K.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2019-03-06</dc:date>
<dc:identifier>doi:10.1101/569095</dc:identifier>
<dc:title><![CDATA[Functional metagenomics-guided discovery of potent Cas9 inhibitors in the human microbiome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/569129v1?rss=1">
<title>
<![CDATA[
A streamlined protocol and analysis pipeline for CUT&RUN chromatin profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/569129v1?rss=1</link>
<description><![CDATA[
We previously described a novel alternative to Chromatin Immunoprecipitation, Cleavage Under Targets & Release Using Nuclease (CUT&RUN), in which unfixed permeabilized cells are incubated with antibody, followed by binding of a Protein A-Micrococcal Nuclease (pA/MNase) fusion protein (1). Upon activation of tethered MNase, the bound complex is excised and released into the supernatant for DNA extraction and sequencing. Here we introduce four enhancements to CUT&RUN: 1) a hybrid Protein A-Protein G-MNase construct that expands antibody compatibility and simplifies purification; 2) a modified digestion protocol that inhibits premature release of the nuclease-bound complex; 3) a calibration strategy based on carry-over of E. coli DNA introduced with the fusion protein; and 4) a novel peak-calling strategy customized for the low-background profiles obtained using CUT&RUN. These new features, coupled with the previously described low-cost, high efficiency, high reproducibility and high-throughput capability of CUT&RUN make it the method of choice for routine epigenomic profiling.
]]></description>
<dc:creator>Meers, M. P.</dc:creator>
<dc:creator>Bryson, T. D.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2019-03-06</dc:date>
<dc:identifier>doi:10.1101/569129</dc:identifier>
<dc:title><![CDATA[A streamlined protocol and analysis pipeline for CUT&RUN chromatin profiling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/587584v1?rss=1">
<title>
<![CDATA[
Quantitation of cis-translational control by general mRNA sequence features in five eukaryotes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/587584v1?rss=1</link>
<description><![CDATA[
BACKGROUNDGeneral translational cis-elements are present in the mRNAs of all genes and affect the recruitment, assembly, and progress of preinitiation complexes and the ribosome under many physiological states. These elements are: mRNA folding, upstream open reading frames, specific nucleotides flanking the initiating AUG codon, protein coding sequence length, and codon usage. The quantitative contributions of these sequence features and how and why they coordinate together to control translation rates are not well understood.nnRESULTSHere we show that these sequence features specify 42%-81% of the variance in translation rates in S. cerevisiae, S. pombe, Arabidopsis thaliana, M. musculus, and H. Sapiens. We establish that control by RNA secondary structure is chiefly mediated by highly folded 25-60 nucleotide segments within mRNA 5 regions; that changes in tri-nucleotide frequencies between highly and poorly translated 5 regions are correlated between all species; and that control by distinct biochemical processes is extensively correlated as is regulation by a single process acting in different parts of the same mRNA.nnCONCLUSIONSOur work shows that the general features control a much larger fraction of the variance in translation rates than previously realized. We provide a more detailed and accurate understanding of the aspects of RNA structure that direct translation in diverse eukaryotes. In addition, we note that the strongly correlated regulation between and within cis-control features will cause more even densities of translational complexes along each mRNA and therefore more efficient use of the translation machinery by the cell.
]]></description>
<dc:creator>Li, J. J.</dc:creator>
<dc:creator>Chew, G.-L.</dc:creator>
<dc:creator>Biggin, M. D.</dc:creator>
<dc:date>2019-03-24</dc:date>
<dc:identifier>doi:10.1101/587584</dc:identifier>
<dc:title><![CDATA[Quantitation of cis-translational control by general mRNA sequence features in five eukaryotes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/590562v1?rss=1">
<title>
<![CDATA[
Orchestrating Single-Cell Analysis with Bioconductor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/590562v1?rss=1</link>
<description><![CDATA[
Recent developments in experimental technologies such as single-cell RNA sequencing have enabled the profiling a high-dimensional number of genome-wide features in individual cells, inspiring the formation of large-scale data generation projects quantifying unprecedented levels of biological variation at the single-cell level. The data generated in such projects exhibits unique characteristics, including increased sparsity and scale, in terms of both the number of features and the number of samples. Due to these unique characteristics, specialized statistical methods are required along with fast and efficient software implementations in order to successfully derive biological insights. Bioconductor - an open-source, open-development software project based on the R programming language - has pioneered the analysis of such high-throughput, high-dimensional biological data, leveraging a rich history of software and methods development that has spanned the era of sequencing. Featuring state-of-the-art computational methods, standardized data infrastructure, and interactive data visualization tools that are all easily accessible as software packages, Bioconductor has made it possible for a diverse audience to analyze data derived from cutting-edge single-cell assays. Here, we present an overview of single-cell RNA sequencing analysis for prospective users and contributors, highlighting the contributions towards this effort made by Bioconductor.
]]></description>
<dc:creator>Amezquita, R. A.</dc:creator>
<dc:creator>Carey, V. J.</dc:creator>
<dc:creator>Carpp, L. N.</dc:creator>
<dc:creator>Geistlinger, L.</dc:creator>
<dc:creator>Lun, A. T.</dc:creator>
<dc:creator>Marini, F.</dc:creator>
<dc:creator>Rue-Albrecht, K.</dc:creator>
<dc:creator>Risso, D.</dc:creator>
<dc:creator>Soneson, C.</dc:creator>
<dc:creator>Waldron, L.</dc:creator>
<dc:creator>Pages, H.</dc:creator>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Huber, W.</dc:creator>
<dc:creator>Morgan, M.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Hicks, S. C.</dc:creator>
<dc:date>2019-03-27</dc:date>
<dc:identifier>doi:10.1101/590562</dc:identifier>
<dc:title><![CDATA[Orchestrating Single-Cell Analysis with Bioconductor]]></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/10.1101/598771v1?rss=1">
<title>
<![CDATA[
Complementing 16S rRNA gene amplicon sequencing with estimates of total bacterial load to infer absolute species concentrations in the vaginal microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/598771v1?rss=1</link>
<description><![CDATA[
Whereas 16S rRNA gene amplicon sequencing quantifies relative abundances of bacterial taxa, variation in total bacterial load between samples restricts its ability to reflect absolute concentration of individual species. Quantitative PCR (qPCR) can quantify individual species, but it is not practical to develop a suite of qPCR assays for every bacterium present in a diverse sample. We analyzed 1320 samples from 20 women with a history of frequent bacterial vaginosis, who self-collected vaginal swabs daily over 60 days. We inferred bacterial concentrations by taking the product of species relative abundance (assessed by 16S rRNA gene amplicon sequencing) and total bacterial load (measured by broad-range 16S rRNA gene qPCR). Log10-converted inferred concentrations correlated with targeted qPCR (r = 0. 935, p<2.2e-16) for seven key bacterial species. The mean inferred concentration error varied across bacteria, with rarer bacterial vaginosis-associated bacteria associated with larger errors. 92% of errors >0.5 log10 occurred when relative abundance was <10%. Many errors occurred during early bacterial expansion or late contraction. When relative abundance of a species is >10%, inferred concentrations are reliable proxies for targeted qPCR. However, targeted qPCR is required to capture bacteria at low relative abundance, particularly with BV-associated bacteria during the early onset of bacterial vaginosis.
]]></description>
<dc:creator>Tettamanti Boshier, F.</dc:creator>
<dc:creator>Srinivasan, S.</dc:creator>
<dc:creator>Lopez, A.</dc:creator>
<dc:creator>Hoffman, N. G.</dc:creator>
<dc:creator>Proll, S.</dc:creator>
<dc:creator>Fredricks, D. N.</dc:creator>
<dc:creator>Schiffer, J. T.</dc:creator>
<dc:date>2019-04-05</dc:date>
<dc:identifier>doi:10.1101/598771</dc:identifier>
<dc:title><![CDATA[Complementing 16S rRNA gene amplicon sequencing with estimates of total bacterial load to infer absolute species concentrations in the vaginal microbiome]]></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/10.1101/610634v1?rss=1">
<title>
<![CDATA[
Elimination of HSV-2 infected cells is mediated predominantly by paracrine effects of tissue-resident T cell derived cytokines 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/610634v1?rss=1</link>
<description><![CDATA[
The mechanisms underlying rapid elimination of herpes simplex virus-2 (HSV-2) in the human genital tract despite low tissue-resident CD8+ T-cell density (TRM) are unknown. We analyzed shedding episodes during chronic HSV-2 infection: viral clearance always occurred within 24 hours of detection even if viral load exceeded 107 HSV DNA copies; surges in granzyme B and interferon-{gamma} occurred within the early hours after reactivation. We next developed a mathematical model of an HSV-2 genital ulcer to integrate mechanistic observations of TRM in situ proliferation, trafficking, cytolytic effects and cytokine alarm signaling from murine studies with viral kinetics, histopathology and lesion size data from humans. A sufficiently high density of HSV-2 specific TRM predicted rapid contact-mediated elimination of infected cells. At lower TRM densities, TRM must initiate a rapidly diffusing, polyfunctional cytokine response in order to eliminate of a majority of infected cells and eradicate briskly spreading HSV-2 infection.nnOne Sentence SummaryControl of herpes simplex virus-2 is primarily mediated by rapidly diffusing cytokines secreted by tissue-resident T cells.
]]></description>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Swan, D. A.</dc:creator>
<dc:creator>Duke, E.</dc:creator>
<dc:creator>Corey, L.</dc:creator>
<dc:creator>Zhu, J.</dc:creator>
<dc:creator>Dave, V.</dc:creator>
<dc:creator>Richert Spuhler, L.</dc:creator>
<dc:creator>Lund, J. M.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:creator>Schiffer, J.</dc:creator>
<dc:date>2019-04-18</dc:date>
<dc:identifier>doi:10.1101/610634</dc:identifier>
<dc:title><![CDATA[Elimination of HSV-2 infected cells is mediated predominantly by paracrine effects of tissue-resident T cell derived cytokines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/629717v1?rss=1">
<title>
<![CDATA[
Thresholds for post-rebound viral control after CCR5 gene-edited autologous hematopoietic cell transplantation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/629717v1?rss=1</link>
<description><![CDATA[
Autologous, CCR5 gene-edited hematopoietic stem and progenitor cell (HSPC) transplantation is a promising strategy for achieving HIV remission. However, only a fraction of HSPCs can be edited ex vivo to provide protection against infection prior to autologous transplantation. The optimal transplantation conditions for achieving viral control in the absence of suppressive antiretroviral therapy (ART) are still unknown. We analyzed data from SHIV-1157ipd3N4-infected juvenile pig-tailed macaques that underwent autologous HSPC transplantation with and without CCR5 gene editing. We developed a mathematical model that recapitulates reconstitution of T cell subset counts and SHIV plasma viral loads in control and transplanted macaques. The model predicts that viral control can be obtained following ART treatment interruption (ATI) when: 1) levels of transplanted HSPCs are at least 10-fold higher than residual endogenous HSPCs after total body irradiation and 2) the fraction of protected HSPCs in the transplant achieves a threshold (73%-90%) sufficient to overcome transplantation-dependent loss of SHIV immunity. Under these conditions, if ATI is withheld until transplanted gene-modified cells engraft and reconstitute to a steady state, then spontaneous viral control is projected to occur immediately. Our results support strategies that 1) increase stem cell dose, 2) enhance potency of conditioning regimen, 3) elevate fraction of gene modified SHIV-resistant cells, 4) extend periods between HSPC transplantation and ATI with tracking of CD4+CCR5- cell recovery and / or 5) augment anti-SHIV immunity to achieve sustained SHIV remission.

One Sentence SummaryAutologous transplantation of {Delta}CCR5 HSPCs may induce post-ATI SHIV control when the gene-edited cell dose is sufficient to overcome SHIV immunity loss.
]]></description>
<dc:creator>Cardozo-Ojeda, E. F.</dc:creator>
<dc:creator>Duke, E. R.</dc:creator>
<dc:creator>Peterson, C. W.</dc:creator>
<dc:creator>Reeves, D. B.</dc:creator>
<dc:creator>Mayer, B. T.</dc:creator>
<dc:creator>Kiem, H.-P.</dc:creator>
<dc:creator>Schiffer, J. T.</dc:creator>
<dc:date>2019-05-07</dc:date>
<dc:identifier>doi:10.1101/629717</dc:identifier>
<dc:title><![CDATA[Thresholds for post-rebound viral control after CCR5 gene-edited autologous hematopoietic cell transplantation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/632166v1?rss=1">
<title>
<![CDATA[
Consensus Machine Learning for Gene Target Selection in Pediatric AML Risk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/632166v1?rss=1</link>
<description><![CDATA[
Acute myeloid leukemia (AML) is a cancer of hematopoietic systems that poses high population burden, especially among pediatric populations. AML presents with high molecular heterogeneity, complicating patient risk stratification and treatment planning. While molecular and cytogenetic subtypes of AML are well described, significance of subtype-specific gene expression patterns is poorly understood and effective modeling of these patterns with individual algorithms is challenging. Using a novel consensus machine learning approach, we analyzed public RNA-seq and clinical data from pediatric AML patients (N = 137 patients) enrolled in the TARGET project.nnWe used a binary risk classifier (Low vs. Not-Low Risk) to study risk-specific expression patterns in pediatric AML. We applied the following workflow to identify important gene targets from RNA-seq data: (1) Reduce data dimensionality by identification of differentially expressed genes for AML risk (N = 1984 loci); (2) Optimize algorithm hyperparameters for each of 4 algorithm types (lasso, XGBoost, random forest, and SVM); (3) Study ablation test results for penalized methods (lasso and XGBoost); (4) Bootstrap Boruta permutations with a novel consensus importance metric.nnWe observed recurrently selected features across hyperparameter optimizations, ablation tests, and Boruta permutation bootstrap iterations, including HOXA9 and putative cofactors including MEIS1. Consensus feature selection from Boruta bootstraps identified a larger gene set than single penalized algorithm runs (lasso or XGBoost), while also including correlated and predictive genes from ablation tests.nnWe present a consensus machine learning approach to identify gene targets of likely importance for pediatric AML risk. The approach identified a moderately sized set of recurrent important genes from across 4 algorithm types, including genes identified across ablation tests with penalized algorithms (HOXA9 and MEIS1). Our approach mitigates exclusion biases of penalized algorithms (lasso and XGBoost) while obviating arbitrary importance cutoffs for other types (SVM and random forest). This approach is readily generalizable for research of other heterogeneous diseases, single-assay experiments, and high-dimensional data. Resources and code to recreate our findings are available online.
]]></description>
<dc:creator>Smith, J.</dc:creator>
<dc:creator>Maden, S.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Buie, R.</dc:creator>
<dc:creator>Peddu, V.</dc:creator>
<dc:creator>Shean, R.</dc:creator>
<dc:creator>Busby, B.</dc:creator>
<dc:date>2019-05-10</dc:date>
<dc:identifier>doi:10.1101/632166</dc:identifier>
<dc:title><![CDATA[Consensus Machine Learning for Gene Target Selection in Pediatric AML Risk]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/643932v1?rss=1">
<title>
<![CDATA[
Impact of rare and common genetic variants on diabetes diagnosis by hemoglobin A1c in multi-ancestry cohorts: The Trans-Omics for Precision Medicine Program. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/643932v1?rss=1</link>
<description><![CDATA[
Hemoglobin A1c (HbA1c) is widely used to diagnose diabetes and assess glycemic control in patients with diabetes. However, nonglycemic determinants, including genetic variation, may influence how accurately HbA1c reflects underlying glycemia. Analyzing the NHLBI Trans-Omics for Precision Medicine (TOPMed) sequence data in 10,338 individuals from five studies and four ancestries (6,158 Europeans, 3,123 African-Americans, 650 Hispanics and 407 East Asians), we confirmed five regions associated with HbA1c (GCK in Europeans and African-Americans, HK1 in Europeans and Hispanics, FN3K/FN3KRP in Europeans and G6PD in African-Americans and Hispanics) and discovered a new African-ancestry specific low-frequency variant (rs1039215 in HBG2/HBE1, minor allele frequency (MAF)=0.03). The most associated G6PD variant (p.Val98Met, rs1050828-T, MAF=12% in African-Americans, MAF=2% in Hispanics) lowered HbA1c (-0.88% in hemizygous males, -0.34% in heterozygous females) and explained 23% of HbA1c variance in African-Americans and 4% in Hispanics. Additionally, we identified a rare distinct G6PD coding variant (rs76723693 - p.Leu353Pro, MAF=0.5%; -0.98% in hemizygous males, -0.46% in heterozygous females) and detected significant association with HbA1c when aggregating rare missense variants in G6PD. We observed similar magnitude and direction of effects for rs1039215 (HBG2) and rs76723693 (G6PD) in the two largest TOPMed African-American cohorts and replicated the rs76723693 association in the UK Biobank African-ancestry participants. These variants in G6PD and HBG2 were monomorphic in the European and Asian samples. African or Hispanic ancestry individuals carrying G6PD variants may be underdiagnosed for diabetes when screened with HbA1c. Thus, assessment of these variants should be considered for incorporation into precision medicine approaches for diabetes diagnosis.
]]></description>
<dc:creator>Sarnowski, C.</dc:creator>
<dc:creator>Leong, A.</dc:creator>
<dc:creator>Raffield, L. M.</dc:creator>
<dc:creator>Wu, P.</dc:creator>
<dc:creator>de Vries, P. S.</dc:creator>
<dc:creator>DiCorpo, D.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Zheng, X.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Brody, J. A.</dc:creator>
<dc:creator>Goodarzi, M. O.</dc:creator>
<dc:creator>Hidalgo, B. A.</dc:creator>
<dc:creator>Highland, H. M.</dc:creator>
<dc:creator>Jain, D.</dc:creator>
<dc:creator>Liu, C.-T.</dc:creator>
<dc:creator>Naik, R. P.</dc:creator>
<dc:creator>Perry, J. A.</dc:creator>
<dc:creator>Porneala, B. C.</dc:creator>
<dc:creator>Selvin, E.</dc:creator>
<dc:creator>Wessel, J.</dc:creator>
<dc:creator>Psaty, B. M.</dc:creator>
<dc:creator>Curran, J. E.</dc:creator>
<dc:creator>Peralta, J. M.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Mathias, R.</dc:creator>
<dc:creator>Johnson, A. D.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Mitchell, B. D.</dc:creator>
<dc:creator>Cupples, L. A.</dc:creator>
<dc:creator>Vasan, R. S.</dc:creator>
<dc:creator>Correa, A.</dc:creator>
<dc:creator>Morrison, A. C.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Rotter, J. I.</dc:creator>
<dc:creator>Rich, S. S.</dc:creator>
<dc:creator>Manning, A. K.</dc:creator>
<dc:creator>Dupuis, J.</dc:creator>
<dc:creator>Meigs, J. B.</dc:creator>
<dc:creator>the TOPMed Diabetes working group,</dc:creator>
<dc:creator>t</dc:creator>
<dc:date>2019-05-28</dc:date>
<dc:identifier>doi:10.1101/643932</dc:identifier>
<dc:title><![CDATA[Impact of rare and common genetic variants on diabetes diagnosis by hemoglobin A1c in multi-ancestry cohorts: The Trans-Omics for Precision Medicine Program.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/648733v1?rss=1">
<title>
<![CDATA[
Reply to \"A discriminative learning approach to differential expression analysis for single-cell RNA-seq\" 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/648733v1?rss=1</link>
<description><![CDATA[
Multivariate logistic regression (mLR) has been recently proposed by Ntranos et al. to perform gene differential expression analyses of single-cell RNA-sequencing (scRNAseq) data. Herein we reproduce and extend some of their findings. We notably show that while mLR performs better in simulated datasets, these simulations do not recapitulate important features of experimental datasets. Indeed, our results suggest that MAST followed by Sidak aggregation of the p-values perform better than mLR on experimental datasets. Overall, we highlight that most of the new results obtained by Ntranos et al is likely due to the quantification of scRNAseq data at the transcript or transcript compatibility classes level, rather than the use of mLR.
]]></description>
<dc:creator>Becht, E.</dc:creator>
<dc:creator>Zhao, E.</dc:creator>
<dc:creator>Amezquita, R. A.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:date>2019-05-30</dc:date>
<dc:identifier>doi:10.1101/648733</dc:identifier>
<dc:title><![CDATA[Reply to \"A discriminative learning approach to differential expression analysis for single-cell RNA-seq\"]]></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/10.1101/654012v1?rss=1">
<title>
<![CDATA[
Multi-ethnic genome-wide association study of decomposed cardioelectric phenotypes illustrates strategies to identify and characterize evidence of shared genetic effects for complex traits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/654012v1?rss=1</link>
<description><![CDATA[
BackgroundPublished genome-wide association studies (GWAS) are mainly European-centric, examine a narrow view of phenotypic variation, and infrequently interrogate genetic effects shared across traits. We therefore examined the extent to which a multi-ethnic, combined trait GWAS of phenotypes that map to well-defined biology can enable detection and characterization of complex trait loci.nnMethodsWith 1000 Genomes Phase 3 imputed data in 34,668 participants (15% African American; 3% Chinese American; 51% European American; 30% Hispanic/Latino), we performed covariate-adjusted univariate GWAS of six contiguous electrocardiogram (ECG) traits that decomposed an average heartbeat and two commonly reported composite ECG traits that summed contiguous traits. Combined phenotype testing was performed using the adaptive sum of powered scores test (aSPU).nnResultsWe identified six novel and 87 known ECG trait loci (aSPU p-value < 5E-9). Lead SNP rs3211938 at novel locus CD36 was common in African Americans (minor allele frequency=10%) and near-monomorphic in European Americans, with effect sizes for the composite trait, QT interval, among the largest reported. Only one novel locus was detected for the composite traits, due to opposite directions of effects across contiguous traits that summed to near-zero. Combined phenotype testing did not detect novel loci unapparent by univariate testing. However, this approach aided locus characterization, particularly when loci harbored multiple independent signals that differed by trait.nnConclusionsDespite including one-third as few participants as the largest published GWAS of ECG traits, our study identifies multiple novel ECG genetic loci, emphasizing the importance of ancestral diversity and phenotype measurement in this era of ever-growing GWAS.nnAUTHOR SUMMARYWe leveraged a multiethnic cohort with precise measures of cardioelectric function to identify novel genetic loci affecting this complex, multifaceted phenotype. The success of our approach stresses the importance of phenotypic precision and participant diversity for future locus discovery and characterization efforts, and cautions against compromises made in genome-wide association studies to pursue ever-growing sample sizes.
]]></description>
<dc:creator>Baldassari, A. R.</dc:creator>
<dc:creator>Avery, C. L.</dc:creator>
<dc:creator>Highland, H. M.</dc:creator>
<dc:creator>Arking, D. R.</dc:creator>
<dc:creator>Buyske, S.</dc:creator>
<dc:creator>Darbar, D.</dc:creator>
<dc:creator>Gondalia, R.</dc:creator>
<dc:creator>Graff, M. R.</dc:creator>
<dc:creator>Guo, X. R.</dc:creator>
<dc:creator>Heckbert, S. R.</dc:creator>
<dc:creator>Hindorff, L. A.</dc:creator>
<dc:creator>Hodonsky, C. J.</dc:creator>
<dc:creator>Ida Chen, Y.-D.</dc:creator>
<dc:creator>Kaplan, R. C.</dc:creator>
<dc:creator>Peters, U.</dc:creator>
<dc:creator>Post, W.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Shohet, R. V.</dc:creator>
<dc:creator>Sotoodehnia, N.</dc:creator>
<dc:creator>Tao, R.</dc:creator>
<dc:creator>Taylor, K. D.</dc:creator>
<dc:creator>Yao, J. D.</dc:creator>
<dc:creator>Kenny, E. D.</dc:creator>
<dc:creator>Lin, H. J.</dc:creator>
<dc:creator>Soliman, E. Z.</dc:creator>
<dc:creator>Whitsel, E. A.</dc:creator>
<dc:creator>North, K. E.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Rotters, j.</dc:creator>
<dc:creator>Seyerle, A. A.</dc:creator>
<dc:date>2019-05-31</dc:date>
<dc:identifier>doi:10.1101/654012</dc:identifier>
<dc:title><![CDATA[Multi-ethnic genome-wide association study of decomposed cardioelectric phenotypes illustrates strategies to identify and characterize evidence of shared genetic effects for complex traits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/662106v1?rss=1">
<title>
<![CDATA[
A new colorectal cancer risk prediction model incorporating family history, personal and environmental factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/662106v1?rss=1</link>
<description><![CDATA[
PurposeReducing colorectal cancer (CRC) incidence and mortality through early detection would improve efficacy if targeted. A CRC risk-prediction model incorporating personal, family, genetic and environmental risk factors could enhance prediction.nnMethodsWe developed risk-prediction models using population-based CRC cases (N=4,445) and controls (N=3,967) recruited by the Colon Cancer Family Registry Cohort (CCFRC). A familial risk profile (FRP) was calculated to summarize individuals risk based on their CRC family history, family structure, germline mutation probability in major susceptibility genes, and a polygenic component. Using logistic regression, we developed risk models including individuals FRP or a binary CRC family-history (FH), and risk factors collected at recruitment. Model validation used follow-up data for population-(N=12,052) and clinic-based (N=5,584) relatives with no cancer history at recruitment, assessing calibration (E/O) and discrimination (AUC).nnResultsThe E/O (95% confidence interval [CI]) for FRP models for population-based relatives were 1.04 (0.74-1.45) and 0.86 (0.64-1.20) for men and women, and for clinic-based relatives 1.15 (0.87-1.58) and 1.04 (0.76-1.45). The age-adjusted AUC (95% CI) for FRP models in population-based relatives were 0.69 (0.60-0.78) and 0.70 (0.62-0.77), and for clinic-based relatives 0.77 (0.69-0.84) and 0.68 (0.60-0.76). The incremental values of AUC (95% CI) for FRP over FH models for population-based relatives were 0.08 (0.01-0.15) and 0.10 (0.04-0.16), and for clinic-based relatives 0.11 (0.05-0.17) and 0.11 (0.06-0.17).nnConclusionThe FRP-based model and FH-based model calibrate well in both settings. The FRP-based model provided better risk-prediction and discrimination than the FH-based model. A detailed family history may be useful for targeted risk-based screening and clinical management.
]]></description>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Hua, X.</dc:creator>
<dc:creator>Win, A. K.</dc:creator>
<dc:creator>MacInnis, R. J.</dc:creator>
<dc:creator>Gallinger, S.</dc:creator>
<dc:creator>Le Marchand, L.</dc:creator>
<dc:creator>Lindor, N. M.</dc:creator>
<dc:creator>Baron, J. A.</dc:creator>
<dc:creator>Hopper, J. L.</dc:creator>
<dc:creator>Dowty, J. G.</dc:creator>
<dc:creator>Antoniou, A. C.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Jenkins, M. A.</dc:creator>
<dc:creator>Newcomb, P. A.</dc:creator>
<dc:date>2019-06-07</dc:date>
<dc:identifier>doi:10.1101/662106</dc:identifier>
<dc:title><![CDATA[A new colorectal cancer risk prediction model incorporating family history, personal and environmental factors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/670497v1?rss=1">
<title>
<![CDATA[
Mapping person-to-person variation in viral mutations that escape polyclonal serum targeting influenza hemagglutinin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/670497v1?rss=1</link>
<description><![CDATA[
A longstanding question is how influenza evolves to escape human immunity, which is polyclonal and can target many distinct epitopes on the virus. Here we map how all amino-acid mutations to influenzas major surface protein affect viral neutralization by polyclonal human sera. The serum of some individuals is so focused that it selects single mutations that reduce viral neutralization by over an order of magnitude. However, different viral mutations escape the sera of different individuals. This individual-to-individual variation in viral escape mutations is not present among ferrets, which are frequently used as a model in influenza studies. Our results show how different single mutations help influenza escape the immunity of different members of the human population, a phenomenon that could shape viral evolution and disease susceptibility.
]]></description>
<dc:creator>Lee, J. M.</dc:creator>
<dc:creator>Eguia, R.</dc:creator>
<dc:creator>Zost, S. J.</dc:creator>
<dc:creator>Choudhary, S.</dc:creator>
<dc:creator>Wilson, P. C.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Stevens-Ayers, T.</dc:creator>
<dc:creator>Boeckh, M.</dc:creator>
<dc:creator>Hurt, A.</dc:creator>
<dc:creator>Lakdawala, S. S.</dc:creator>
<dc:creator>Hensley, S. E.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2019-06-13</dc:date>
<dc:identifier>doi:10.1101/670497</dc:identifier>
<dc:title><![CDATA[Mapping person-to-person variation in viral mutations that escape polyclonal serum targeting influenza hemagglutinin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/677559v1?rss=1">
<title>
<![CDATA[
Persistence and plasticity of T follicular memory cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/677559v1?rss=1</link>
<description><![CDATA[
CD4 memory T cells play an important role in protective immunity and are a key target in vaccine development. Many studies have focused on T central memory (TCM) cells, while the existence and functional significance of T follicular helper (TFH) memory cells is controversial. Here we show that TFH memory cells are highly susceptible to NAD induced cell death (NICD) during isolation from tissues, leading to their under-representation in prior studies. NICD blockade reveals the persistence of abundant TFH memory cells, with high expression of hallmark TFH markers, that persist to at least 400 days after infection, by which time TCM cells are no longer found. Using single cell RNA-seq we demonstrate that TFH memory cells are transcriptionally distinct from TCM cells, maintain stemness and self-renewal gene expression, and, in contrast to TCM cells, are multipotent following recall. Surprisingly, TFH memory cells concurrently express a distinct glycolytic signature similar to trained immune cells, including elevated expression of mTOR, HIF-1 and cAMP regulated genes. Late disruption of glycolysis/ICOS signaling leads to TFH memory cell depletion concomitant with decreased splenic plasma cells and circulating antibody titers, demonstrating both unique homeostatic regulation of memory TFH and their sustained function during the memory phase of the immune response. These results highlight the metabolic heterogeneity underlying distinct memory T cell subsets and establish TFH memory cells as an attractive target for the induction of long-lived adaptive immunity.nnHIGHLIGHTSO_LICell death during isolation from the tissue prevents the full recovery of TFH memory cellsnC_LIO_LITFH memory cells are transcriptionally distinct from TCM cells and maintain a broader recall capacitynC_LIO_LITFH memory cells are maintained in the absence of antigen but require ICOS signaling and glycolysisnC_LIO_LITFH memory cells support late phase antibody production by splenic plasma cellsnC_LI
]]></description>
<dc:creator>Künzli, M.</dc:creator>
<dc:creator>Schreiner, D. P.</dc:creator>
<dc:creator>Pereboom, T.</dc:creator>
<dc:creator>Swarnalekha, N.</dc:creator>
<dc:creator>Lötscher, J.</dc:creator>
<dc:creator>Ertuna, Y. I.</dc:creator>
<dc:creator>Roux, J.</dc:creator>
<dc:creator>Geier, F.</dc:creator>
<dc:creator>Hess, C.</dc:creator>
<dc:creator>Taylor, J. T.</dc:creator>
<dc:creator>King, C. G.</dc:creator>
<dc:date>2019-06-23</dc:date>
<dc:identifier>doi:10.1101/677559</dc:identifier>
<dc:title><![CDATA[Persistence and plasticity of T follicular memory cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/680611v1?rss=1">
<title>
<![CDATA[
Pulmonary natural killer cells control neutrophil intravascular motility and response to acute inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/680611v1?rss=1</link>
<description><![CDATA[
The pulmonary immune system defends a huge surface area directly in contact with the contents of the air we breathe. Neutrophils, the most abundant immune cell in the pulmonary vasculature, are critical to immunity but they are also capable of generating life-threatening pathology. Natural Killer cells are the most highly represented lymphocyte subset in the lung, but relatively little is known about their localization, motility or the specific mechanisms by which they contribute to local homeostasis. Here, we used lung-intravital microscopy to directly visualise and quantify neutrophil and natural killer cell dynamics in the pulmonary vasculature of live mice. This approach revealed unexpected sessile behaviour by intravascular natural killer cells. Interactions with natural killer cells made neutrophils scan the endothelium more slowly over larger distances and reduced the number of neutrophils that accumulated in an LPS-triggered inflammatory challenge. This represents a new paradigm by which natural killer cells contribute to lung physiology by diminishing potentially pathogenic neutrophil accumulation.
]]></description>
<dc:creator>Secklehner, J.</dc:creator>
<dc:creator>De Filippo, K.</dc:creator>
<dc:creator>Mackey, J. B. G.</dc:creator>
<dc:creator>Vuononvirta, J.</dc:creator>
<dc:creator>Raffo Iraolagoitia, X. L.</dc:creator>
<dc:creator>McFarlane, A. J.</dc:creator>
<dc:creator>Neilson, M.</dc:creator>
<dc:creator>Headley, M. B.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:creator>Guerra, N.</dc:creator>
<dc:creator>Carlin, L.</dc:creator>
<dc:date>2019-06-23</dc:date>
<dc:identifier>doi:10.1101/680611</dc:identifier>
<dc:title><![CDATA[Pulmonary natural killer cells control neutrophil intravascular motility and response to acute inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/687236v1?rss=1">
<title>
<![CDATA[
Trans- and cis-acting effects of the lncRNA Firre on epigenetic and structural features of the inactive X chromosome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/687236v1?rss=1</link>
<description><![CDATA[
Firre encodes a lncRNA involved in nuclear organization in mammals. Here we find that Firre RNA is transcribed from the active X chromosome (Xa) and exerts trans-acting effects on the inactive X chromosome (Xi). Allelic deletion of Firre on the Xa in a mouse hybrid fibroblast cell line results in a dramatic loss of the histone modification H3K27me3 and of components of the PRC2 complex on the Xi as well as the disruption of the perinucleolar location of the Xi. These features are measurably rescued by ectopic expression of a mouse or human Firre/FIRRE cDNA transgene, strongly supporting a conserved trans-acting role of the Firre transcript in maintaining the Xi heterochromatin environment. Surprisingly, CTCF occupancy is decreased on the Xi upon loss of Firre RNA, but is partially recovered by ectopic transgene expression, suggesting a functional link between Firre RNA and CTCF in maintenance of epigenetic features and/or location of the Xi. Loss of Firre RNA results in dysregulation of genes implicated in cell division and development, but not in reactivation of genes on the Xi, which retains its bipartite structure despite some changes in chromatin contact distribution. Allelic deletion or inversion of Firre on the Xi causes localized redistribution of chromatin contacts, apparently dependent on the orientation of CTCF binding sites clustered at the locus. Thus, the Firre locus and its RNA have roles in the maintenance of epigenetic features and structure of the Xi.
]]></description>
<dc:creator>FANG, H.</dc:creator>
<dc:creator>Bonora, G.</dc:creator>
<dc:creator>Lewandowski, J.</dc:creator>
<dc:creator>Thakur, J.</dc:creator>
<dc:creator>Filippova, G. N.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Duan, Z.</dc:creator>
<dc:creator>Rinn, J.</dc:creator>
<dc:creator>Deng, X.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:date>2019-06-30</dc:date>
<dc:identifier>doi:10.1101/687236</dc:identifier>
<dc:title><![CDATA[Trans- and cis-acting effects of the lncRNA Firre on epigenetic and structural features of the inactive X chromosome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/696278v1?rss=1">
<title>
<![CDATA[
Partial absence of PD-1 expression by tumor-specific CD8+ T cells in EBV-driven lymphoepithelioma-like carcinoma: a case report 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/696278v1?rss=1</link>
<description><![CDATA[
Lymphoepithelioma-like carcinoma (LELC) is an uncommon lung cancer, typically observed in young, non-smoking Asian populations. LELC is associated with Epstein-Barr virus (EBV) infection of lung tumor cells of epithelial origin, suggesting a carcinogenic role of EBV as observed in nasopharyngeal carcinoma (NPC). Here, we studied the antigen specificity and phenotype of CD8+ tumor infiltrating lymphocytes (TILs) in one LELC patient positive for EBV infection in lung tumor cells. Using MHC class I tetramers, we detected two populations of EBV-specific CD8+ TILs, which can be considered as tumor-specific CD8+ T cells, in the tumor of this patient. Transcriptomic analyses of these two populations reveal their distinct exhausted profiles and polyclonal TCR repertoire. High dimensional analyses at single cell level using mass cytometry showed showed that populations of tumor specific CD8+ TILs are phenotypically heterogeneous, although they consistently express CD39. Unexpectedly, although the LELC tumor cells expressed abundant PD-L1, these tumor-specific CD8+ TILs mostly did not express PD-1, suggesting that anti-PD1/PD-L1 immunotherapy may not be an appropriate strategy for disinhibiting EBV-specific cells for the treatment of LELC patients. These results might also help to explain low rates of checkpoint blockade immunotherapy response for NPC, despite the antigenicity of EBV for both tumor types.
]]></description>
<dc:creator>SIMONI, Y.</dc:creator>
<dc:creator>Becht, E.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>LOH, C. Y.</dc:creator>
<dc:creator>YEONG, J. P. S.</dc:creator>
<dc:creator>LIM, T. K. H.</dc:creator>
<dc:creator>TAKANO, A.</dc:creator>
<dc:creator>TAN, D. S. W.</dc:creator>
<dc:creator>NEWELL, E. W.</dc:creator>
<dc:date>2019-07-09</dc:date>
<dc:identifier>doi:10.1101/696278</dc:identifier>
<dc:title><![CDATA[Partial absence of PD-1 expression by tumor-specific CD8+ T cells in EBV-driven lymphoepithelioma-like carcinoma: a case report]]></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/10.1101/700534v1?rss=1">
<title>
<![CDATA[
A targeted multi-omic analysis approach measures protein expression and low abundance transcripts on the single cell level 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/700534v1?rss=1</link>
<description><![CDATA[
High throughput single-cell RNA sequencing (sc-RNAseq) has become a frequently used tool to assess immune cell function and heterogeneity. Recently, the combined measurement of RNA and protein expression by sequencing was developed, which is commonly known as CITE-Seq. Acquisition of protein expression data along with transcriptome data resolves some of the limitations inherent to only assessing transcript, but also nearly doubles the sequencing read depth required per single cell. Furthermore, there is still a paucity of analysis tools to visualize combined transcript-protein datasets.nnHere, we describe a novel targeted transcriptomics approach that combines analysis of over 400 genes with simultaneous measurement of over 40 proteins on more than 25,000 cells. This targeted approach requires only about 1/10 of the read depth compared to a whole transcriptome approach while retaining high sensitivity for low abundance transcripts. To analyze these multi-omic transcript-protein datasets, we adapted One-SENSE for intuitive visualization of the relationship of proteins and transcripts on a single-cell level.
]]></description>
<dc:creator>Mair, F.</dc:creator>
<dc:creator>Erickson, J. R.</dc:creator>
<dc:creator>Voillet, V.</dc:creator>
<dc:creator>Simoni, Y.</dc:creator>
<dc:creator>Bi, T.</dc:creator>
<dc:creator>Tyznik, A. J.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:date>2019-07-14</dc:date>
<dc:identifier>doi:10.1101/700534</dc:identifier>
<dc:title><![CDATA[A targeted multi-omic analysis approach measures protein expression and low abundance transcripts on the single cell level]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/701961v1?rss=1">
<title>
<![CDATA[
Oral tenofovir disoproxil fumarate/emtricitabine for HIV pre-exposure prophylaxis increases expression of type I/III interferon-stimulated factors in the gastrointestinal tract but not in the blood 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/701961v1?rss=1</link>
<description><![CDATA[
Tenofovir disoproxil fumarate (TDF) and emtricitabine (FTC) are used for HIV treatment and prevention. Previously, we found that topical rectal tenofovir gel caused immunological changes in the mucosa. Here we assessed the effect of oral TDF/FTC in three HIV pre-exposure prophylaxis trials, two with gastrointestinal and one with cervicovaginal biopsies. TDF/FTC induced type I/III interferon-related (IFN I/III) genes in the gastrointestinal tract, but not blood, with strong correlations between the two independent rectal biopsy groups (Spearman r=0.91) and between the rectum and duodenum (r=0.81). Gene set testing also indicated stimulation of type I/III pathways in the ectocervix, as well as of cellular proliferation in the duodenum. mRNA sequencing, digital droplet PCR, proteomics, and immunofluorescence staining confirmed IFN I/III pathway stimulation in the gastrointestinal tract. Thus, oral TDF/FTC stimulates an IFN-I/III signature throughout the gut, which could increase antiviral efficacy but also cause chronic immune activation in HIV prevention and treatment settings.
]]></description>
<dc:creator>Hughes, S. M.</dc:creator>
<dc:creator>Levy, C. N.</dc:creator>
<dc:creator>Calienes, F.</dc:creator>
<dc:creator>Stekler, J. D.</dc:creator>
<dc:creator>Pandey, U.</dc:creator>
<dc:creator>Vojtech, L.</dc:creator>
<dc:creator>Berard, A. R.</dc:creator>
<dc:creator>Birse, K.</dc:creator>
<dc:creator>Noel-Romas, L.</dc:creator>
<dc:creator>Richardson, B.</dc:creator>
<dc:creator>Golden, J.</dc:creator>
<dc:creator>Cartwright, M.</dc:creator>
<dc:creator>Collier, A. C.</dc:creator>
<dc:creator>Stevens, C. E.</dc:creator>
<dc:creator>Curlin, M. E.</dc:creator>
<dc:creator>Holtz, T. H.</dc:creator>
<dc:creator>Mugo, N.</dc:creator>
<dc:creator>Irungu, E.</dc:creator>
<dc:creator>Katabira, E.</dc:creator>
<dc:creator>Muwonge, T.</dc:creator>
<dc:creator>Lama, J. R.</dc:creator>
<dc:creator>Baeten, J. M.</dc:creator>
<dc:creator>Burgener, A.</dc:creator>
<dc:creator>Lingappa, J. R.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Mackelprang, R.</dc:creator>
<dc:creator>McGowan, I.</dc:creator>
<dc:creator>Cranston, R. D.</dc:creator>
<dc:creator>Cameron, M. J.</dc:creator>
<dc:creator>Hladik, F.</dc:creator>
<dc:date>2019-07-13</dc:date>
<dc:identifier>doi:10.1101/701961</dc:identifier>
<dc:title><![CDATA[Oral tenofovir disoproxil fumarate/emtricitabine for HIV pre-exposure prophylaxis increases expression of type I/III interferon-stimulated factors in the gastrointestinal tract but not in the blood]]></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/10.1101/702118v1?rss=1">
<title>
<![CDATA[
A new data-driven cell population discovery and annotation method for single-cell data, FAUST, reveals correlates of clinical response to cancer immunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/702118v1?rss=1</link>
<description><![CDATA[
High-dimensional single-cell cytometry is routinely used to characterize patient responses to cancer immunotherapy and other treatments. This has produced a wealth of datasets ripe for exploration but whose biological and technical heterogeneity make them difficult to analyze with current tools. We introduce a new interpretable machine learning method for single-cell mass and flow cytometry studies, FAUST, that robustly performs unbiased cell population discovery and annotation. FAUST processes data on a per-sample basis and returns biologically interpretable cell phenotypes that can be compared across studies, making it well-suited for the analysis and integration of complex datasets. We demonstrate how FAUST can be used for candidate biomarker discovery and validation by applying it to a flow cytometry dataset from a Merkel cell carcinoma anti-PD-1 trial and discover new CD4+ and CD8+ effector-memory T cell correlates of outcome co-expressing PD-1, HLA-DR, and CD28. We then use FAUST to validate these correlates in an independent CyTOF dataset from a published metastatic melanoma trial. Importantly, existing state-of-the-art computational discovery approaches as well as prior manual analysis did not detect these or any other statistically significant T cell sub-populations associated with anti-PD-1 treatment in either data set. We further validate our methodology by using FAUST to replicate the discovery of a previously reported myeloid correlate in a different published melanoma trial, and validate the correlate by identifying it de novo in two additional independent trials. FAUSTs phenotypic annotations can be used to perform cross-study data integration in the presence of heterogeneous data and diverse immunophenotyping staining panels, enabling hypothesis-driven inference about cell sub-population abundance through a multivariate modeling framework we call Phenotypic and Functional Differential Abundance (PFDA). We demonstrate this approach on data from myeloid and T cell panels across multiple trials. Together, these results establish FAUST as a powerful and versatile new approach for unbiased discovery in single-cell cytometry.
]]></description>
<dc:creator>Greene, E.</dc:creator>
<dc:creator>Finak, G.</dc:creator>
<dc:creator>D'Amico, L. A.</dc:creator>
<dc:creator>Bhardwaj, N.</dc:creator>
<dc:creator>Church, C. D.</dc:creator>
<dc:creator>Morishima, C.</dc:creator>
<dc:creator>Ramchurren, N.</dc:creator>
<dc:creator>Taube, J. M.</dc:creator>
<dc:creator>Nghiem, P. T.</dc:creator>
<dc:creator>Cheever, M. A.</dc:creator>
<dc:creator>Fling, S. P.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:date>2019-07-13</dc:date>
<dc:identifier>doi:10.1101/702118</dc:identifier>
<dc:title><![CDATA[A new data-driven cell population discovery and annotation method for single-cell data, FAUST, reveals correlates of clinical response to cancer immunotherapy]]></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/10.1101/722009v1?rss=1">
<title>
<![CDATA[
The Effects of Genotype x Phenotype Interactions on Silver Nanoparticle Toxicity in Organotypic Cultures of Murine Tracheal Epithelial Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/722009v1?rss=1</link>
<description><![CDATA[
Silver nanoparticles (AgNP) are used in multiple applications but primarily in the manufacturing of antimicrobial products. Previous studies have identified AgNP toxicity in airway epithelial cells, but no in vitro studies to date have used organotypic cultures as a high-content in vitro model of the conducting airway to characterize the effects of interactions between host genetic and acquired factors, or gene x phenotype interactions (GxP), on AgNP toxicity. In the present study, we derived organotypic cultures from primary murine tracheal epithelial cells (MTEC) to characterize nominal and dosimetric dose-response relationships for AgNP-induced barrier dysfunction, glutathione (GSH) depletion, reactive oxygen species (ROS) production, lipid peroxidation, and cytotoxicity across two genotypes (A/J and C57BL/6J mice), two phenotypes ("Normal" and "Type 2 [T2]-Skewed"), and two exposures (an acute exposure of 24 h and a subacute exposure of 4 hours, every other day, over 5 days [5x4 h]). We characterized the "T2-Skewed" phenotype as an in vitro model of chronic respiratory diseases, which was marked by increased sensitivity to AgNP-induced barrier dysfunction, GSH depletion, ROS production, lipid peroxidation, and cytotoxicity, suggesting that asthmatics are a sensitive population to AgNP exposures in occupational settings. This also suggests that exposure limits, which should be based upon the most sensitive population, should be derived using in vitro and in vivo models of chronic respiratory diseases. This study highlights the importance of considering dosimetry as well as GxP effects when screening and prioritizing potential respiratory toxicants. Such in vitro studies can be used to inform regulatory policy aimed at special protections for all populations.
]]></description>
<dc:creator>Nicholas, T. P.</dc:creator>
<dc:creator>Haick, A. K.</dc:creator>
<dc:creator>Workman, T. W.</dc:creator>
<dc:creator>Griffith, W. C.</dc:creator>
<dc:creator>Nolin, J. D.</dc:creator>
<dc:creator>Kavanagh, T. J.</dc:creator>
<dc:creator>Faustman, E. M.</dc:creator>
<dc:creator>Altemeier, W. A.</dc:creator>
<dc:date>2019-08-01</dc:date>
<dc:identifier>doi:10.1101/722009</dc:identifier>
<dc:title><![CDATA[The Effects of Genotype x Phenotype Interactions on Silver Nanoparticle Toxicity in Organotypic Cultures of Murine Tracheal Epithelial 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/10.1101/725556v1?rss=1">
<title>
<![CDATA[
Deep mutational scanning comprehensively maps how Zika envelope protein mutations affect viral growth and antibody escape 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/725556v1?rss=1</link>
<description><![CDATA[
Functional constraints on viral proteins are often assessed by examining sequence conservation among natural strains, but this approach is relatively ineffective for Zika virus because all known sequences are highly similar. Here we take an alternative approach to map functional constraints on Zika viruss envelope (E) protein by using deep mutational scanning to measure how all amino-acid mutations to the protein affect viral growth in cell culture. The resulting sequence-function map is consistent with existing knowledge about E protein structure and function, but also provides insight into mutation-level constraints in many regions of the protein that have not been well characterized in prior functional work. In addition, we extend our approach to completely map how mutations affect viral neutralization by two monoclonal antibodies, thereby precisely defining their functional epitopes. Overall, our study provides a valuable resource for understanding the effects of mutations to this important viral protein, and also offers a roadmap for future work to map functional and antigenic selection to Zika virus at high resolution.nnImportanceZika virus has recently been shown to be associated with severe birth defects. The viruss E protein mediates its ability to infect cells, and is also the primary target of the antibodies that are elicited by natural infection and vaccines that are being developed against the virus. Therefore, determining the effects of mutations to this protein is important for understanding its function, its susceptibility to vaccine-mediated immunity, and its potential for future evolution. We completely mapped how amino-acid mutations to E protein affected the viruss ability to grow in cells in the lab and escape from several antibodies. The resulting maps relate changes in the E proteins sequence to changes in viral function, and therefore provide a valuable complement to existing maps of the physical structure of the protein.
]]></description>
<dc:creator>Sourisseau, M.</dc:creator>
<dc:creator>Lawrence, D. J. P.</dc:creator>
<dc:creator>Schwarz, M. C.</dc:creator>
<dc:creator>Storrs, C. H.</dc:creator>
<dc:creator>Veit, E. C.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Evans, M. J.</dc:creator>
<dc:date>2019-08-05</dc:date>
<dc:identifier>doi:10.1101/725556</dc:identifier>
<dc:title><![CDATA[Deep mutational scanning comprehensively maps how Zika envelope protein mutations affect viral growth and antibody escape]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/725903v1?rss=1">
<title>
<![CDATA[
INX-18 and INX-19 play distinct roles in electrical synapses that modulate aversive behavior in Caenorhabditis elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/725903v1?rss=1</link>
<description><![CDATA[
In order to respond to changing environments and fluctuations in internal states, animals adjust their behavior through diverse neuromodulatory mechanisms. In this study we show that electrical synapses between the ASH primary quinine-detecting sensory neurons and the neighboring ASK neurons are required for modulating the aversive response to the bitter tastant quinine in C. elegans. Mutant worms that lack the electrical synapse proteins INX-18 and INX-19 become hypersensitive to dilute quinine. Cell-specific rescue experiments indicate that inx-18 operates in ASK while inx-19 is required in both ASK and ASH for proper quinine sensitivity. Imaging analyses find that INX-19 in ASK and ASH localizes to the same regions in the nerve ring, suggesting that both sides of ASK-ASH electrical synapses contain INX-19. While inx-18 and inx-19 mutant animals have a similar behavioral phenotype, several lines of evidence suggest the proteins encoded by these genes play different roles in modulating the aversive quinine response. First, INX-18 and INX-19 localize to different regions of the nerve ring, indicating that they are not present in the same synapses. Second, removing inx-18 disrupts the distribution of INX-19, while removing inx-19 does not alter INX-18 localization. Finally, by using a fluorescent cGMP reporter, we find that INX-18 and INX-19 have distinct roles in establishing cGMP levels in ASK and ASH. Together, these results demonstrate that electrical synapses containing INX-18 and INX-19 facilitate modulation of ASH nociceptive signaling. Our findings support the idea that a network of electrical synapses mediates cGMP exchange between neurons, enabling modulation of sensory responses and behavior.nnAuthor SummaryAnimals are constantly adjusting their behavior to respond to changes in the environment or to their internal state. This behavior modulation is achieved by altering the activity of neurons and circuits through a variety of neuroplasticity mechanisms. Chemical synapses are known to impact neuroplasticity in several different ways, but the diversity of mechanisms by which electrical synapses contribute is still being investigated. Electrical synapses are specialized sites of connection between neurons where ions and small signaling molecules can pass directly from one cell to the next. By passing small molecules through electrical synapses, neurons may be able to modify the activity of their neighbors. In this study we identify two genes that contribute to electrical synapses between two sensory neurons in C. elegans. We show that these electrical synapses are crucial for proper modulation of sensory responses, as without them animals are overly responsive to an aversive stimulus. In addition to pinpointing their sites of action, we present evidence that they may be contributing to neuromodulation by facilitating passage of the small molecule cGMP between neurons. Our work provides evidence for a role of electrical synapses in regulating animal behavior.
]]></description>
<dc:creator>Voelker, L.</dc:creator>
<dc:creator>Upadhyaya, B.</dc:creator>
<dc:creator>Ferkey, D. M.</dc:creator>
<dc:creator>Woldemariam, S.</dc:creator>
<dc:creator>L'Etoile, N. D.</dc:creator>
<dc:creator>Rabinowitch, I.</dc:creator>
<dc:creator>Bai, J.</dc:creator>
<dc:date>2019-08-05</dc:date>
<dc:identifier>doi:10.1101/725903</dc:identifier>
<dc:title><![CDATA[INX-18 and INX-19 play distinct roles in electrical synapses that modulate aversive behavior in Caenorhabditis elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/734541v1?rss=1">
<title>
<![CDATA[
DUX4-induced bidirectional HSATII satellite repeat transcripts form intranuclear double stranded RNA foci in human cell models of FSHD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/734541v1?rss=1</link>
<description><![CDATA[
The DUX4 transcription factor is normally expressed in the cleavage stage embryo and regulates genes involved in embryonic genome activation. Mis-expression of DUX4 in skeletal muscle, however, is toxic and causes facioscapulohumeral muscular dystrophy (FSHD). We recently showed DUX4-induced toxicity is due, in part, to the activation of the double-stranded RNA (dsRNA) response pathway and the accumulation of intranuclear dsRNA foci. Here, we determined the composition of DUX4-induced dsRNAs. We found that a subset of DUX4-induced dsRNAs originate from inverted Alu repeats embedded within the introns of DUX4-induced transcripts and from DUX4-induced dsRNA-forming intergenic transcripts enriched for endogenous retroviruses, Alu and LINE-1 elements. However, these repeat classes were also represented in dsRNAs from cells not expressing DUX4. In contrast, pericentric human satellite II (HSATII) repeats formed a class of dsRNA specific to the DUX4 expressing cells. Further investigation revealed that DUX4 can initiate the bidirectional transcription of normally heterochromatin-silenced HSATII repeats. DUX4 induced HSATII RNAs co-localized with DUX4-induced nuclear dsRNA foci and with intranuclear aggregation of EIF4A3 and ADAR1. Finally, gapmer-mediated knockdown of HSATII transcripts depleted DUX4-induced intranuclear ribonucleoprotein aggregates and decreased DUX4-induced cell death, suggesting that HSATII formed dsRNAs contribute to DUX4 toxicity.
]]></description>
<dc:creator>Shadle, S. C.</dc:creator>
<dc:creator>Bennett, S. R.</dc:creator>
<dc:creator>Wong, C.-J.</dc:creator>
<dc:creator>Karreman, N. A.</dc:creator>
<dc:creator>Campbell, A. E.</dc:creator>
<dc:creator>van der Maarel, S. M.</dc:creator>
<dc:creator>Bass, B. L.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:date>2019-08-13</dc:date>
<dc:identifier>doi:10.1101/734541</dc:identifier>
<dc:title><![CDATA[DUX4-induced bidirectional HSATII satellite repeat transcripts form intranuclear double stranded RNA foci in human cell models of FSHD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/737742v1?rss=1">
<title>
<![CDATA[
Chromosomal Mcm2-7 distribution is a primary driver of genome replication timing in budding yeast, fission yeast and mouse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/737742v1?rss=1</link>
<description><![CDATA[
The spatio-temporal program of genome replication across eukaryotes is thought to be driven both by the uneven loading of pre-replication complexes (pre-RCs) across the genome at the onset of S-phase, and by differences in the timing of activation of these complexes during S-phase. To determine the degree to which distribution of pre-RC loading alone could account for chromosomal replication patterns, we mapped the binding sites of the Mcm2-7 helicase complex (MCM) in budding yeast, fission yeast, mouse and humans. We observed identical MCM double-hexamer footprints across the species, but notable differences in their distribution: In budding yeast, complexes were present in sharp peaks comprised largely of single double-hexamers; in fission yeast, corresponding peaks typically contained 4 to 8 double-hexamers, were more disperse, and showed a striking correlation with AT content. In mouse and humans, complexes were even more disperse, with a preference for regions of high GC content. Nonetheless, most fluctuations in replication timing in all four organisms could be accounted for by differences in chromosomal MCM distribution. This analysis also identified genomic regions whose replication timing was clearly not attributable to MCM density. The most notable was the inactive X-chromosome, which replicates late in S phase despite the fact that both MCM abundance and chromosomal distribution were comparable to those on the early replicating active X-chromosome. We conclude that, although certain genomic regions, most notably the inactive X-chromosome, are subject to post-licensing regulation, most differences in replication timing along the chromosome reflect uneven chromosomal distribution of stochastically firing pre-replication complexes.
]]></description>
<dc:creator>Foss, E. J.</dc:creator>
<dc:creator>Sripathy, S.</dc:creator>
<dc:creator>Gatobonton-Schwager, T.</dc:creator>
<dc:creator>Kwak, H.</dc:creator>
<dc:creator>Thiesen, A. H.</dc:creator>
<dc:creator>Lao, U.</dc:creator>
<dc:creator>Bedalov, A.</dc:creator>
<dc:date>2019-08-16</dc:date>
<dc:identifier>doi:10.1101/737742</dc:identifier>
<dc:title><![CDATA[Chromosomal Mcm2-7 distribution is a primary driver of genome replication timing in budding yeast, fission yeast and mouse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/741025v1?rss=1">
<title>
<![CDATA[
Inherited chromosomally integrated HHV-6 demonstrates tissue-specific RNA expression in vivo that correlates with increased antibody immune response. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/741025v1?rss=1</link>
<description><![CDATA[
Human herpesvirus-6A and 6B (HHV-6A, HHV-6B) are human viruses capable of chromosomal integration. Approximately 1% of the human population carry one copy of HHV-6A/B integrated into every cell in their body, referred to as inherited chromosomally integrated human herpesvirus 6A/B (iciHHV-6A/B). Whether iciHHV-6A/B is transcriptionally active in vivo and how it shapes the immunological response is still unclear. Here, we screened DNA-Seq and RNA-Seq data for 650 individuals available through the Genotype-Tissue Expression (GTEx) project and identified 2 iciHHV-6A and 4 iciHHV-6B positive candidates. When corresponding tissue-specific gene expression signatures were analyzed, low levels HHV-6A/B gene expression was found across multiple tissues, with the highest levels of gene expression in the brain (specifically for iciHHV-6A), testis, esophagus, and adrenal gland. U90 and U100 were the most highly expressed HHV-6 genes in both iciHHV-6A and iciHHV-6B individuals. To assess whether tissue-specific gene expression from iciHHV-6A/B influences the immune response, a cohort of 15,498 subjects was screened and 85 iciHHV-6A/B+ subjects were identified. Plasma samples from iciHHV-6A/B+ and age and sex matched controls were analyzed for antibodies to control antigens (CMV, EBV, FLU) or HHV-6A/B antigens. Our results indicate that iciHHV-6A/B+ subjects have significantly more antibodies against the U90 gene product (IE1) relative to non-iciHHV-6 individuals. Antibody responses against EBV and FLU antigens or HHV-6A/B gene products either not expressed or expressed at low levels, such as U47, U57 or U72, were identical between controls and iciHHV-6A/B+ subjects. CMV seropositive individuals with iciHHV-6A/B+ have more antibodies against CMV pp150, relative to CMV seropositive controls. These results argue that spontaneous gene expression from integrated HHV-6A/B leads to an increase in antigenic burden that translates into a more robust HHV-6A/B specific antibody response.nnImportanceHHV-6A/B are human herpesviruses that have the unique property of being able to integrate into the subtelomeric regions of human chromosomes. Approximately 1% of the worlds population carries integrated HHV-6A/B genome in every cell of their body. Whether viral genes are transcriptionally active in these individuals is unclear. By taking advantage of a unique tissue-specific gene expression dataset, we show the majority of tissues from iciHHV-6 individuals do not show HHV-6 gene expression. Brain and testes showed the highest tissue-specific expression of HHV-6 genes in two separate datasets. Two HHV-6 genes, U90 (immediate early 1 protein) and U100 (glycoproteins Q1 and Q2), were found to be selectively and consistently expressed across several human tissues. Expression of U90 translates into an increase in antigen-specific antibody response in iciHHV-6A/B+ subjects relative to controls. Future studies will be needed to determine the mechanism of gene expression, the effects of these genes on human gene transcription networks and the pathophysiological impact of having increased viral protein expression in tissue in conjunction with increased antigen-specific antibody production.
]]></description>
<dc:creator>Peddu, V.</dc:creator>
<dc:creator>Dubuc, I.</dc:creator>
<dc:creator>Gravel, A.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Tenenbaum, D.</dc:creator>
<dc:creator>Jerome, K.</dc:creator>
<dc:creator>Tardif, J.-C.</dc:creator>
<dc:creator>Dube, M.-P.</dc:creator>
<dc:creator>Flamand, L.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:date>2019-08-21</dc:date>
<dc:identifier>doi:10.1101/741025</dc:identifier>
<dc:title><![CDATA[Inherited chromosomally integrated HHV-6 demonstrates tissue-specific RNA expression in vivo that correlates with increased antibody immune response.]]></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/10.1101/745059v1?rss=1">
<title>
<![CDATA[
Two separate roles for the transcription coactivator SAGA and a set of genes redundantly regulated by TFIID and SAGA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/745059v1?rss=1</link>
<description><![CDATA[
Deletions within genes coding for subunits of the transcription coactivator SAGA caused strong genome-wide defects in transcription and SAGA-mediated chromatin modifications. In contrast, rapid SAGA depletion produced only modest transcription defects at 13% of protein-coding genes - genes that are generally more sensitive to rapid TFIID depletion. However, transcription of these "coactivator-redundant" genes is strongly affected by rapid depletion of both factors, showing the overlapping functions of TFIID and SAGA at this gene set. We suggest that this overlapping function is linked to TBP-DNA recruitment. The remaining 87% of expressed genes that we term "TFIID-dependent" are highly sensitive to rapid TFIID depletion and insensitive to rapid SAGA depletion. Genome-wide mapping of SAGA and TFIID found binding of both factors at many genes independent of gene class. DNA analysis suggests that the distinction between the gene classes is due to multiple components rather than any single regulatory factor or promoter sequence motif.
]]></description>
<dc:creator>Donczew, R.</dc:creator>
<dc:creator>Warfield, L.</dc:creator>
<dc:creator>Erijman, A.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:date>2019-08-22</dc:date>
<dc:identifier>doi:10.1101/745059</dc:identifier>
<dc:title><![CDATA[Two separate roles for the transcription coactivator SAGA and a set of genes redundantly regulated by TFIID and SAGA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/748434v1?rss=1">
<title>
<![CDATA[
Sensitive detection of highly fragmented cytomegalovirus nucleic acid in human cfDNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/748434v1?rss=1</link>
<description><![CDATA[
Congenital human cytomegalovirus (CMV) infections are the leading cause of newborn hearing and central nervous system impairments worldwide. Currently, routine prenatal screening for congenital CMV is not performed in the United States and confirmation in suspected perinatal cases requires invasive sampling by amniocentesis. We hypothesized that detection of CMV from maternal cell-free DNA (cfDNA) plasma could provide a non-invasive indicator of congenital CMV infection. We analyzed sequence data from 2,208 individuals undergoing routine non-invasive prenatal aneuploidy screening at the University of Washington. CMV reads were identified in 117 (5.3%) samples. Positive samples were stratified based on CMV reads per million sample reads (RPM), resulting in ten samples being classified as strong positive (RPM > 0.3) and 107 as intermediate positive (0.01<RPM<0.3). Subsequent qPCR testing identified CMV in 9/10 strong positive samples and 2/32 intermediate positive samples. Median cfDNA insert size derived from CMV was significantly shorter than cfDNA derived from human chromosomes (103 vs 172 bp, p<0.0001), corresponding to the 3rd percentile of human cfDNA insert size. In addition, CMV cfDNA fragment lengths were distributed over a wider range than human cfDNA reads. These studies reveal the highly fragmented nature of CMV cfDNA and offer precise measurements of its length: these features likely explain discrepancies in serum CMV viral loads measurements determined by different qPCR assays, despite widespread efforts to standardize results. More work is required to determine how detection of CMV from maternal cfDNA can be best used as tool for congenital CMV screening or diagnosis.
]]></description>
<dc:creator>Peddu, V.</dc:creator>
<dc:creator>Bradley, B.</dc:creator>
<dc:creator>Shree, S.</dc:creator>
<dc:creator>Colbert, B.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Santo, T.</dc:creator>
<dc:creator>Huang, M.</dc:creator>
<dc:creator>Cheng, E.</dc:creator>
<dc:creator>Konnick, E.</dc:creator>
<dc:creator>Salipante, S.</dc:creator>
<dc:creator>Jerome, K.</dc:creator>
<dc:creator>Lockwood, C.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:date>2019-08-28</dc:date>
<dc:identifier>doi:10.1101/748434</dc:identifier>
<dc:title><![CDATA[Sensitive detection of highly fragmented cytomegalovirus nucleic acid in human cfDNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/762484v1?rss=1">
<title>
<![CDATA[
Physical activity and risks of breast and colorectal cancer: A Mendelian randomization analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/762484v1?rss=1</link>
<description><![CDATA[
Physical activity has been associated with lower risks of breast and colorectal cancer in epidemiological studies; however, it is unknown if these associations are causal or confounded. In two-sample Mendelian randomization analyses, using summary genetic data from the UK Biobank and GWA consortia, we found that a one standard deviation increment in average acceleration was associated with lower risks of breast cancer (odds ratio [OR]: 0.59, 95% confidence interval [CI]: 0.42 to 0.84, P-value=0.003) and colorectal cancer (OR: 0.66, 95% CI: 0.53 to 0.82, P-value=2*E-4). We found similar magnitude inverse associations by breast cancer subtype and by colorectal cancer anatomical site. Our results support a potentially causal relationship between higher physical activity levels and lower risks of breast cancer and colorectal cancer. Based on these data, the promotion of physical activity is probably an effective strategy in the primary prevention of these commonly diagnosed cancers.nnDisclaimerWhere authors are identified as personnel of the International Agency for Research on Cancer / World Health Organization, the authors alone are responsible for the views expressed in this article and they do not necessarily represent the decisions, policy or views of the International Agency for Research on Cancer / World Health Organization.
]]></description>
<dc:creator>Papadimitriou, N.</dc:creator>
<dc:creator>Dimou, N.</dc:creator>
<dc:creator>Tsilidis, K. K.</dc:creator>
<dc:creator>Banbury, B.</dc:creator>
<dc:creator>Martin, R.</dc:creator>
<dc:creator>Lewis, S.</dc:creator>
<dc:creator>Kazmi, N.</dc:creator>
<dc:creator>Robinson, T. M.</dc:creator>
<dc:creator>Albanes, D.</dc:creator>
<dc:creator>Aleksandrova, K.</dc:creator>
<dc:creator>Berndt, S. I.</dc:creator>
<dc:creator>Bishop, D. T.</dc:creator>
<dc:creator>Brenner, H.</dc:creator>
<dc:creator>Buchanan, D. D.</dc:creator>
<dc:creator>Bueno-de-Mesquita, B.</dc:creator>
<dc:creator>Campbell, P. T.</dc:creator>
<dc:creator>Castellvi-Bel, S.</dc:creator>
<dc:creator>Chan, A. T.</dc:creator>
<dc:creator>Chang-Claude, J.</dc:creator>
<dc:creator>Ellingjord-Dale, M.</dc:creator>
<dc:creator>Figueiredo, J. C.</dc:creator>
<dc:creator>Gallinger, S. J.</dc:creator>
<dc:creator>Giles, G. G.</dc:creator>
<dc:creator>Giovannucci, E.</dc:creator>
<dc:creator>Gruber, S. B.</dc:creator>
<dc:creator>Gsur, A.</dc:creator>
<dc:creator>Hampe, J.</dc:creator>
<dc:creator>Hampel, H.</dc:creator>
<dc:creator>Harlid, S.</dc:creator>
<dc:creator>Harrison, T. A.</dc:creator>
<dc:creator>Hoffmeister, M.</dc:creator>
<dc:creator>Hopper, J. L.</dc:creator>
<dc:creator>Hsu, L.</dc:creator>
<dc:creator>Huerta, J. M.</dc:creator>
<dc:creator>Huyghe, J. R.</dc:creator>
<dc:creator>Jenkins, M. A.</dc:creator>
<dc:creator>Keku, T. O.</dc:creator>
<dc:creator>Kuhn, T.</dc:creator>
<dc:creator>La Vecchia, C.</dc:creator>
<dc:creator>Le Marchand, L.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2019-09-09</dc:date>
<dc:identifier>doi:10.1101/762484</dc:identifier>
<dc:title><![CDATA[Physical activity and risks of breast and colorectal cancer: A Mendelian randomization analysis]]></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/10.1101/765313v1?rss=1">
<title>
<![CDATA[
New mutant mouse models clarify the role of NAIPs, phosphorylation, NLRP3, and tumors in NLRC4 inflammasome activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/765313v1?rss=1</link>
<description><![CDATA[
The NAIP/NLRC4 inflammasome is a cytosolic sensor of bacteria that activates Caspase-1 and initiates potent downstream immune responses. Structural, biochemical, and genetic data all demonstrate that the NAIP proteins act as receptors for specific bacterial ligands, while NLRC4 is a downstream adaptor protein that multimerizes with NAIPs to form a macromolecular structure called an inflammasome. However, several aspects of NLRC4 biology remain unresolved. For example, in addition to its clear function in responding to bacteria, NLRC4 has also been proposed to initiate anti-tumor responses, though the underlying mechanism is unknown. NLRC4 has also been shown to be phosphorylated on serine 533, and this modification was suggested to be important for NLRC4 function. In the absence of S533 phosphorylation, it was further proposed that another inflammasome component, NLRP3, can induce NLRC4 activation. We generated a new Nlrc4-deficient mouse line as well as mice encoding phosphomimetic S533D and non-phosphorylatable S533A NLRC4 proteins. Using these genetic models in vivo and in vitro, we fail to observe a role for phosphorylation in NLRC4 inflammasome function. Furthermore, we find no role for NLRP3 in NLRC4 function, or for NLRC4 in a model of melanoma. These results simplify and clarify our understanding of the mechanism of NAIP/NLRC4 activation and its biological functions.
]]></description>
<dc:creator>Tenthorey, J.</dc:creator>
<dc:creator>Chavez, R. A.</dc:creator>
<dc:creator>Thompson, T. W.</dc:creator>
<dc:creator>Deets, K. A.</dc:creator>
<dc:creator>Vance, R. E.</dc:creator>
<dc:creator>Rauch, I.</dc:creator>
<dc:date>2019-09-18</dc:date>
<dc:identifier>doi:10.1101/765313</dc:identifier>
<dc:title><![CDATA[New mutant mouse models clarify the role of NAIPs, phosphorylation, NLRP3, and tumors in NLRC4 inflammasome activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/769711v1?rss=1">
<title>
<![CDATA[
Progressive differentiation of memory CD8 T cells in the cervicovaginal tissue 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/769711v1?rss=1</link>
<description><![CDATA[
Tissue-resident memory CD8 T cells (CD8 TRM) are critical for maintaining barrier immunity. CD8 TRM have been mainly studied in the skin and gut with recent studies suggesting that the signals that control tissue-residence and phenotype are highly tissue-dependent. We examined the T cell compartment in healthy human cervicovaginal tissue (CVT) and found that most CD8 T cells were granzyme B+ and TCF-1-. To address if this phenotype is driven by CVT tissue-residence, we used a mouse model to control for environmental factors. Using localized and systemic infection models, we found that CD8 TRM in the mouse CVT gradually acquired a granzyme B+, TCF-1- phenotype as seen in human CVT. In contrast to CD8 TRM in the gut, these CD8 TRM were not stably maintained regardless of the initial infection route, which led to reductions in local immunity. Our data show that residence in the CVT is sufficient to progressively shape the size and function of its CD8 TRM compartment.

SummaryThe tissue-resident memory (TRM) CD8 T cell compartment in human and mouse cervicovaginal tissue (CVT) is remarkably similar. The CVT TRM compartment is maintained autonomously and does not reach phenoypical or numerical equilibrium. The numerical decline leads to impaired viral control in a secondary challenge.
]]></description>
<dc:creator>Dave, V.</dc:creator>
<dc:creator>Cardozo Ojeda, E. F.</dc:creator>
<dc:creator>Mair, F.</dc:creator>
<dc:creator>Erickson, J.</dc:creator>
<dc:creator>Woodward-Davis, A.</dc:creator>
<dc:creator>Soerens, A.</dc:creator>
<dc:creator>Czartoski, J.</dc:creator>
<dc:creator>Teague, C.</dc:creator>
<dc:creator>Oberle, S.</dc:creator>
<dc:creator>Zehn, D.</dc:creator>
<dc:creator>Schiffer, J.</dc:creator>
<dc:creator>Lund, J. M.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:date>2019-09-14</dc:date>
<dc:identifier>doi:10.1101/769711</dc:identifier>
<dc:title><![CDATA[Progressive differentiation of memory CD8 T cells in the cervicovaginal tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/776856v1?rss=1">
<title>
<![CDATA[
Diversity and function of maternal HIV-1-specific antibodies at the time of vertical transmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/776856v1?rss=1</link>
<description><![CDATA[
Infants of HIV positive mothers can acquire HIV infection by various routes, but even in the absence of antiviral treatment, the majority of these infants do not become infected. There is evidence that maternal antibodies may provide some protection from infection, but gestational maternal antibodies have not yet been characterized in detail. One of the most studied vertically-infected infants is BG505, as the virus from this infant yielded an Envelope protein that was successfully developed as a stable trimer. Here, we isolated and characterized 39 HIV-specific neutralizing monoclonal antibodies (nAbs) from MG505, the mother of BG505, at a time point just prior to vertical transmission. These nAbs belonged to 21 clonal families, employed a variety of VH genes, many were specific for the HIV-1 Env V3 loop, and this V3 specificity correlated with measurable antibody-dependent cellular cytotoxicity (ADCC) activity. The isolated nAbs did not recapitulate the full breadth of heterologous nor autologous virus neutralization by contemporaneous plasma. Notably, we found that the V3-targeting nAb families neutralized one particular maternal Env variant even though all tested variants had low V3 sequence diversity and were measurably bound by these nAbs. None of the nAbs neutralized the BG505 transmitted virus. Furthermore, the MG505 nAb families were found at relatively low frequencies within the maternal B cell repertoire: all less than 0.25% of total IgG sequences. Our findings demonstrate the diversity of HIV-1 nAbs that exist within a single mother, resulting in a collection of antibody specificities that can shape the transmission bottleneck.nnImportanceMother-to-child-transmission of HIV-1 offers a unique setting in which maternal antibodies both within the mother and passively-transferred to the infant are present at the time of viral exposure. Untreated HIV-exposed human infants are infected at a rate of 30-40%, meaning that some infants do not get infected despite continued exposure to virus. Since the potential of HIV-specific immune responses to provide protection against HIV is a central goal of HIV vaccine design, understanding the nature of maternal antibodies may provide insights into immune mechanisms of protection. In this study, we isolated and characterized HIV-specific antibodies from the mother of an infant whose transmitted virus has been well studied.
]]></description>
<dc:creator>Doepker, L.</dc:creator>
<dc:creator>Simonich, C.</dc:creator>
<dc:creator>Ralph, D.</dc:creator>
<dc:creator>Gobillot, T. A.</dc:creator>
<dc:creator>Garrett, M.</dc:creator>
<dc:creator>Vigdorovich, V.</dc:creator>
<dc:creator>Sather, D. N.</dc:creator>
<dc:creator>Nduati, R.</dc:creator>
<dc:creator>Matsen, F.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2019-09-23</dc:date>
<dc:identifier>doi:10.1101/776856</dc:identifier>
<dc:title><![CDATA[Diversity and function of maternal HIV-1-specific antibodies at the time of vertical transmission]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/780627v1?rss=1">
<title>
<![CDATA[
Seasonal influenza circulation patterns and projections for September 2019 to September 2020 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/780627v1?rss=1</link>
<description><![CDATA[
This report details current seasonal influenza circulation patterns as of August 2019 and makes projections up to September 2020 to coincide with selection of the 2020 Southern Hemisphere vaccine strain. This is not meant as a comprehensive report, but is instead intended as particular observations that weve made that may be of relevance. Please also note that observed patterns reflect the GISAID database and may not be entirely representative of underlying dynamics. All analyses are based on the nextflu/nextstrain pipeline [1,2] with continual updates posted to nextstrain.org/flu. A/H3N2: A/H3N2 viruses continue to show substantial diversity in HA sequences with a deep split between 3c3.A and 3c2.A1b viruses. The most notable recent developments are the rapid rise of clade A1b/137F - a subclade of A1b/135K - in China and Bangladesh and clade A1b/197R - a subclade of A1b/131K - which dominates the ongoing season in Australia. Our models predict that A1b/137F and A1b/197R will be the dominant clades next year with A1b/197R accounting for most circulation. There is, however, large uncertainty in the true extent of A1b/137F circulation. A/H1N1pdm: The S183P substitution has risen to near fixation. The most successful subclade carrying this mutation is 183P-5 which has essentially replaced competing variants. A variant with substitutions 129D/185I is at 60% prevalence globally, while a second variant with substitution 130N is at 50% in North America and ~10% elsewhere. Substitutions at site 156 to D or K have arisen sporadically and result in loss of recognition by antisera raised against viruses with asparagine at position 156. Despite the large antigenic effect, viruses with mutations at site 156 dont seem to spread. Beyond variants at site 156, little to no antigenic evolution is evident in assays with ferret antisera. B/Vic: Antigenically drifted deletion variants at HA1 sites 162, 163 and 164 are now dominating global circulation and have all but taken over. The double deletion variant V1A.1 had previously been circulating at high frequency in the Americas. However, over the course of 2009, the triple deletion variant V1A.3 has increased in frequency globally and is now dominating in all geographic regions. Importantly, V1A.1 and V1A.3 variants appear antigenically distinct by HI assays with 4-8 fold reductions in log2 titer in both directions. B/Yam: B/Yam has not circulated in large numbers since the Northern Hemisphere season 2017/2018 and displays relatively little amino acid variation in HA or antigenic diversity. Amino acid variants at sites 229 and 232 have begun to circulate and population is now split between 229D/232D, 229N/232D and 229D/232N variants. These variants show little sign of antigenic difference in HI assays.
]]></description>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Potter, B.</dc:creator>
<dc:creator>Neher, R.</dc:creator>
<dc:date>2019-09-24</dc:date>
<dc:identifier>doi:10.1101/780627</dc:identifier>
<dc:title><![CDATA[Seasonal influenza circulation patterns and projections for September 2019 to September 2020]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/781526v1?rss=1">
<title>
<![CDATA[
Adipokines set neural tone by regulating synapse number 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/781526v1?rss=1</link>
<description><![CDATA[
Energy sensing neural circuits decide to expend or conserve resources by integrating tonic steady-state energy store information with phasic signals for hunger and food intake. Tonic signals, in the form of adipose tissue-derived adipokines, set the baseline level of energy-sensing neuron activity, providing context for interpretation of phasic messages. However, the mechanism by which tonic adipokine information establishes baseline neuronal function is unclear. Here we show that Upd2, a Drosophila Leptin ortholog, regulates actin-based synapse reorganization by reducing inhibitory synaptic contacts, thereby providing a permissive neural tone for insulin release under conditions of nutrient surplus. Unexpectedly, Insulin acts on the same upstream inhibitory neurons to conversely increase synapse number, hence re-instating negative tone. Our results suggest that two surplus-sensing hormonal systems, Leptin/Upd2 and Insulin, converge on a neuronal circuit with opposing outcomes that establish tonic, energy-store-dependent neuron activity.

HighlightsO_LIThe adipokine Upd2 regulates number of inhibitory synaptic contacts on Insulin neurons.
C_LIO_LIUpd2 activates an actin-regulating complex of Arouser, Basigin, and Gelsolin in target neurons.
C_LIO_LIArouser, Basigin, and Gelsolin reduce the extent of inhibitory contact on Insulin neurons.
C_LIO_LIInsulin resets negative tone by increasing the number of synaptic contacts made by its own upstream inhibitory neurons.
C_LI
]]></description>
<dc:creator>Brent, A. E.</dc:creator>
<dc:creator>Rajan, A.</dc:creator>
<dc:date>2019-09-25</dc:date>
<dc:identifier>doi:10.1101/781526</dc:identifier>
<dc:title><![CDATA[Adipokines set neural tone by regulating synapse number]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-25</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/10.1101/783118v1?rss=1">
<title>
<![CDATA[
A High-Resolution Landscape of Mutations in the BCL6 Super-Enhancer in Normal Human B-Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/783118v1?rss=1</link>
<description><![CDATA[
The super-enhancers (SE) of lineage-specific genes in B-cells are off-target sites of somatic hypermutation. However, the inability to detect sufficient numbers of mutations in normal human B-cells has precluded the generation of a high-resolution mutational landscape of SEs. Here, we captured and sequenced 12 B-cell SEs at single-nucleotide resolution from ten healthy individuals across diverse ethnicities. We detected a total of [~]9000 subclonal mutations (allele frequencies <0.1%); of these, [~]8000 are present in the BCL6 SE alone. Within the BCL6 SE, we identified three regions of clustered mutations where the mutation frequency is [~]7X10-4. Mutational spectra show a predominance of C>T/G>A and A>G/T>C substitutions, consistent with the activities of activation-induced-cytidine deaminase (AID) and the A-T mutator, DNA Polymerase {eta}, respectively, in mutagenesis in normal B-cells. Analyses of mutational signatures further corroborate the participation of these factors in this process. Single base substitution signature SBS85, SBS37, and SBS39 were found in the BCL6 SE. While SBS85 is a denoted signature of AID in lymphoid cells, the etiologies of SBS37 and SBS39 are still unknown. Our analysis suggests the contribution of error-prone DNA polymerases to the latter signatures. The high-resolution mutation landscape has enabled accurate profiling of subclonal mutations in B-cell SEs in normal individuals. By virtue of the fact that subclonal SE mutations are clonally expanded in B-cell lymphomas, our studies also offer the potential for early detection of neoplastic alterations.nnSignificanceWe used Duplex Sequencing to detect low-frequency mutations in the BCL6 super-enhancer locus in normal human B-cells. The landscape of pre-existing mutations is remarkably conserved across different ethnicities and reveals clustered mutational hotspots that correlate with reported sites of clonal mutations and translocation breakpoints in human B-cell lymphomas. This high-resolution genomic landscape revealed by Duplex Sequencing offers accurate and thorough profiling of low frequency, pre-existing mutations in normal individuals, and the potential for early detection of neoplastic alterations.
]]></description>
<dc:creator>Shen, J.-C.</dc:creator>
<dc:creator>Kamath-Loeb, A. S.</dc:creator>
<dc:creator>Kohrn, B. F.</dc:creator>
<dc:creator>Loeb, K. R.</dc:creator>
<dc:creator>Preston, B. D.</dc:creator>
<dc:creator>Loeb, L. A.</dc:creator>
<dc:date>2019-09-26</dc:date>
<dc:identifier>doi:10.1101/783118</dc:identifier>
<dc:title><![CDATA[A High-Resolution Landscape of Mutations in the BCL6 Super-Enhancer in Normal Human B-Cells]]></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/10.1101/784835v1?rss=1">
<title>
<![CDATA[
Architectural RNA is required for heterochromatin organization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/784835v1?rss=1</link>
<description><![CDATA[
In addition to its known roles in protein synthesis and enzyme catalysis, RNA has been proposed to stabilize higher-order chromatin structure. To distinguish presumed architectural roles of RNA from other functions, we applied a ribonuclease digestion strategy to our CUT&RUN in situ chromatin profiling method (CUT&RUN.RNase). We find that depletion of RNA compromises association of the murine nucleolar protein Nucleophosmin with pericentric heterochromatin and alters the chromatin environment of CCCTC-binding factor (CTCF) bound regions. Strikingly, we find that RNA maintains the integrity of both constitutive (H3K9me3 marked) and facultative (H3K27me3 marked) heterochromatic regions as compact domains, but only moderately stabilizes euchromatin. To establish the specificity of heterochromatin stabilization by RNA, we performed CUT&RUN on cells deleted for the Firre long non-coding RNA and observed disruption of H3K27me3 domains on several chromosomes. We conclude that RNA maintains local and global chromatin organization by acting as a structural scaffold for heterochromatic domains.
]]></description>
<dc:creator>Thakur, J.</dc:creator>
<dc:creator>Fang, H.</dc:creator>
<dc:creator>Llagas, T.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2019-09-27</dc:date>
<dc:identifier>doi:10.1101/784835</dc:identifier>
<dc:title><![CDATA[Architectural RNA is required for heterochromatin organization]]></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/10.1101/790642v1?rss=1">
<title>
<![CDATA[
Functional characterization and lineage analysis of broadly neutralizing human antibodies against dengue virus identified by single B cell transcriptomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/790642v1?rss=1</link>
<description><![CDATA[
Eliciting broadly neutralizing antibodies (bNAbs) against the four dengue virus serotypes (DENV1-4) that are spreading into new territories is an important goal of vaccine design. To delineate bNAb targets, we characterized 28 monoclonal antibodies belonging to expanded and hypermutated clonal families identified by transcriptomic analysis of single plasmablasts from DENV-infected individuals. Among these, we identified two somatically related bNAbs that potently neutralized DENV1-4. Mutagenesis studies revealed that the major recognition determinants of these bNAbs are in E protein domain I, distinct from the only known class of human bNAbs against flaviviruses with a well-defined epitope. B cell repertoire analysis from acute-phase peripheral blood suggested a memory origin and divergent somatic hypermutation pathways for these bNAbs, and a limited number of mutations was sufficient for neutralizing activity. Our study suggests multiple B cell evolutionary pathways leading to DENV bNAbs targeting a novel epitope that can be exploited for vaccine design.
]]></description>
<dc:creator>Durham, N. D.</dc:creator>
<dc:creator>Agrawal, A.</dc:creator>
<dc:creator>Waltari, E.</dc:creator>
<dc:creator>Croote, D.</dc:creator>
<dc:creator>Zanini, F.</dc:creator>
<dc:creator>Fouch, M.</dc:creator>
<dc:creator>Davidson, E.</dc:creator>
<dc:creator>Smith, O.</dc:creator>
<dc:creator>Carabajal, E.</dc:creator>
<dc:creator>Pak, J. E.</dc:creator>
<dc:creator>Doranz, B. J.</dc:creator>
<dc:creator>Robinson, M.</dc:creator>
<dc:creator>Sanz, A. M.</dc:creator>
<dc:creator>Albornoz, L. L.</dc:creator>
<dc:creator>Rosso, F.</dc:creator>
<dc:creator>Einav, S.</dc:creator>
<dc:creator>Quake, S. R.</dc:creator>
<dc:creator>McCutcheon, K. M.</dc:creator>
<dc:creator>Goo, L.</dc:creator>
<dc:date>2019-10-02</dc:date>
<dc:identifier>doi:10.1101/790642</dc:identifier>
<dc:title><![CDATA[Functional characterization and lineage analysis of broadly neutralizing human antibodies against dengue virus identified by single B cell transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/808394v1?rss=1">
<title>
<![CDATA[
When Two are Better than One: Modeling the Mechanisms of Antibody Mixtures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/808394v1?rss=1</link>
<description><![CDATA[
It is difficult to predict how antibodies will behave when mixed together, even after each has been independently characterized. Here, we present a statistical mechanical model for the activity of antibody mixtures that accounts for whether pairs of antibodies bind to distinct or overlapping epitopes. This model requires measuring n individual antibodies and their [Formula] pairwise interactions to predict the 2n potential combinations. We apply this model to epidermal growth factor receptor (EGFR) antibodies and find that the activity of antibody mixtures can be predicted without positing synergy at the molecular level. In addition, we demonstrate how the model can be used in reverse, where straightforward experiments measuring the activity of antibody mixtures can be used to infer the molecular interactions between antibodies. Lastly, we generalize this model to analyze engineered multidomain antibodies, where components of different antibodies are tethered together to form novel amalgams, and characterize how well it predicts recently designed influenza antibodies.

Author summaryWith the rise of new antibody combinations in therapeutic regimens, it is important to understand how antibodies work together as well as individually. Here, we investigate the specific case of monoclonal antibodies targeting a cancer-causing receptor or the influenza virus and develop a statistical mechanical framework that predicts the effectiveness of a mixture of antibodies. The power of this model lies in its ability to make a large number of predictions based on a limited amount of data. For example, once 10 antibodies have been individually characterized and their epitopes have been mapped, our model can predict how any of the 210 = 1024 combinations will behave. This predictive power can aid therapeutic efforts by assessing which combinations of antibodies will elicit the most effective response.
]]></description>
<dc:creator>Einav, T.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2019-10-17</dc:date>
<dc:identifier>doi:10.1101/808394</dc:identifier>
<dc:title><![CDATA[When Two are Better than One: Modeling the Mechanisms of Antibody Mixtures]]></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/10.1101/812016v1?rss=1">
<title>
<![CDATA[
Linking influenza virus evolution within and between human hosts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/812016v1?rss=1</link>
<description><![CDATA[
Influenza viruses rapidly diversify within individual human infections. Several recent studies have deep-sequenced clinical influenza infections to identify viral variation within hosts, but it remains unclear how within-host mutations fare in the global viral population. Here, we compare viral variation within and between hosts to link influenzas evolutionary dynamics across scales. Synonymous sites evolve at similar rates at both scales, indicating that global evolution at these putatively neutral sites results from the accumulation of within-host variation. However, nonsynonymous mutations are depleted in global viral populations compared to within hosts, suggesting that selection purges many of the protein-altering changes that arise within hosts. The exception is at antigenic sites, where selection detectably favors nonsynonymous mutations at the global scale, but not within hosts. These results suggest that selection against deleterious mutations and selection for antigenic change are the main forces that transform influenzas within-host genetic variation into global evolution.
]]></description>
<dc:creator>Xue, K. S.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2019-10-21</dc:date>
<dc:identifier>doi:10.1101/812016</dc:identifier>
<dc:title><![CDATA[Linking influenza virus evolution within and between human hosts]]></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/10.1101/814145v1?rss=1">
<title>
<![CDATA[
A comparative study of topology-based pathway enrichment analysis methods 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/814145v1?rss=1</link>
<description><![CDATA[
BackgroundPathway enrichment analysis is extensively used in the analysis of Omics data for gaining biological insights into the functional roles of pre-defined subsets of genes, proteins and metabolites. A large number of methods have been proposed in the literature for this task. The vast majority of these methods use as input expression levels of the biomolecules under study together with their membership in pathways of interest. The latest generation of pathway enrichment methods also leverages information on the topology of the underlying pathways, which as evidence from their evaluation reveals, lead to improved sensitivity and specificity. Nevertheless, a systematic empirical comparison of such methods is still lacking, making selection of the most suitable method for a specific experimental setting challenging. This comparative study of nine network-based methods for pathway enrichment analysis aims to provide a systematic evaluation of their performance based on three real data sets with different number of features (genes/metabolites) and number of samples.nnResultsThe findings highlight both methodological and empirical differences across the nine methods. In particular, certain methods assess pathway enrichment due to differences both across expression levels and in the strength of the interconnectedness of the members of the pathway, while others only leverage differential expression levels. In the more challenging setting involving a metabolomics data set, the results show that methods that utilize both pieces of information (with NetGSA being a prototypical one) exhibit superior statistical power in detecting pathway enrichment.nnConclusionThe analysis reveals that a number of methods perform equally well when testing large size pathways, which is the case with genomic data. On the other hand, NetGSA that takes into consideration both differential expression of the biomolecules in the pathway, as well as changes in the topology exhibits a superior performance when testing small size pathways, which is usually the case for metabolomics data.
]]></description>
<dc:creator>Ma, J.</dc:creator>
<dc:creator>Shojaie, A.</dc:creator>
<dc:creator>Michailidis, G.</dc:creator>
<dc:date>2019-10-22</dc:date>
<dc:identifier>doi:10.1101/814145</dc:identifier>
<dc:title><![CDATA[A comparative study of topology-based pathway enrichment analysis methods]]></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/10.1101/814269v1?rss=1">
<title>
<![CDATA[
The GMD-biplot and its application to microbiome data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/814269v1?rss=1</link>
<description><![CDATA[
Exploratory analysis of human microbiome data is often based on dimension-reduced graphical displays derived from similarities based on non-Euclidean distances, such as UniFrac or Bray-Curtis. However, a display of this type, often referred to as the principal coordinate analysis (PCoA) plot, does not reveal which taxa are related to the observed clustering because the configuration of samples is not based on a coordinate system in which both the samples and variables can be represented. The reason is that the PCoA plot is based on the eigen-decomposition of a similarity matrix and not the singular value decomposition (SVD) of the sample-by-abundance matrix. We propose a novel biplot that is based on an extension of the SVD, called the generalized matrix decomposition (GMD), which involves an arbitrary matrix of similarities and the original matrix of variable measures, such as taxon abundances. As in a traditional biplot, points represent the samples and arrows represent the variables. The proposed GMD-biplot is illustrated by analyzing multiple real and simulated data sets which demonstrate that the GMD-biplot provides improved clustering capability and a more meaningful relationship between the arrows and the points.
]]></description>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Randolph, T.</dc:creator>
<dc:creator>Shojaie, A.</dc:creator>
<dc:creator>Ma, J.</dc:creator>
<dc:date>2019-10-22</dc:date>
<dc:identifier>doi:10.1101/814269</dc:identifier>
<dc:title><![CDATA[The GMD-biplot and its application to microbiome data]]></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/10.1101/820886v1?rss=1">
<title>
<![CDATA[
TRIM34 acts with TRIM5 to restrict HIV and SIV capsids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/820886v1?rss=1</link>
<description><![CDATA[
The HIV-1 capsid protein makes up the core of the virion and plays a critical role in early steps of HIV replication. Due to its exposure in the cytoplasm after entry, HIV capsid is a target for host cell factors that act directly to block infection such as TRIM5 and MxB. Several host proteins also play a role in facilitating infection, including in the protection of HIV-1 capsid from recognition by host cell restriction factors. Through an unbiased screening approach, called HIV-CRISPR, we show that the Cyclophilin A-binding deficient P90A HIV-1 capsid mutant becomes highly-sensitized to TRIM5alpha restriction in IFN-treated cells. Further, the CPSF6-binding deficient, N74D HIV-1 capsid mutant is sensitive to restriction mediated by human TRIM34, a close paralog of the well-characterized HIV restriction factor TRIM5. This restriction occurs at the step of reverse transcription, is independent of interferon stimulation and limits HIV-1 infection in key target cells of HIV infection including CD4+ T cells and monocyte-derived dendritic cells. TRIM34 restriction requires TRIM5alpha as knockout or knockdown of TRIM5alpha results in a loss of antiviral activity. TRIM34 can also restrict some SIV capsids. Through immunofluorescence studies, we show that TRIM34 and TRIM5alpha colocalize to cytoplasmic bodies and are more frequently observed to be associated with infecting N74D capsids than with WT capsids. Our results identify TRIM34 as an HIV-1 CA-targeting restriction factor and highlight the potential role for heteromultimeric TRIM interactions in contributing restriction of HIV-1 infection in human cells.
]]></description>
<dc:creator>Ohainle, M.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Keceli, S.</dc:creator>
<dc:creator>Felton, A.</dc:creator>
<dc:creator>Campbell, E. M.</dc:creator>
<dc:creator>Luban, J.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2019-10-30</dc:date>
<dc:identifier>doi:10.1101/820886</dc:identifier>
<dc:title><![CDATA[TRIM34 acts with TRIM5 to restrict HIV and SIV capsids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/826735v1?rss=1">
<title>
<![CDATA[
Retinoic acid organizes the vagus motor topographic map via spatiotemporal regulation of Hgf/Met signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/826735v1?rss=1</link>
<description><![CDATA[
The topographic map, in which the positions of neuron cell bodies correspond with the positions of their synaptic targets, is a major organizational motif in the nervous system. To understand how topographic axon targeting is controlled during development, we examine the mechanism underlying topographic innervation of the pharyngeal arches by the vagus motor nerve in zebrafish. We reveal that Retinoic Acid organizes the topographic map by specifying anterior-posterior identity in post-mitotic vagus motor neurons. We then show that chemoattractant signaling between hepatocyte growth factor (Hgf) and the Met receptor is required for pharyngeal arch innervation by the vagus motor nerve. Finally, we find that Retinoic Acid controls the spatiotemporal dynamics of Hgf/Met signaling to coordinate axon targeting with the developmental progression of the pharyngeal arches and show that experimentally altering the timing of Hgf/Met signaling is sufficient to redirect axon targeting and disrupt the topographic map. These findings establish a new mechanism of topographic map development in which regulation of chemoattractant signaling in both space and time guides axon targeting.
]]></description>
<dc:creator>Isabella, A. J.</dc:creator>
<dc:creator>Barsh, G. R.</dc:creator>
<dc:creator>Stonick, J. A.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:date>2019-10-31</dc:date>
<dc:identifier>doi:10.1101/826735</dc:identifier>
<dc:title><![CDATA[Retinoic acid organizes the vagus motor topographic map via spatiotemporal regulation of Hgf/Met signaling]]></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/10.1101/827055v1?rss=1">
<title>
<![CDATA[
Detection and treatment strategy for Tritrichomonas muris in the common laboratory mouse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/827055v1?rss=1</link>
<description><![CDATA[
Maintaining a specific pathogen-free (SPF) mouse colony is critical to avoid potentially confounding variables introduced by unknown infections. Here, we report an instance of protozoan Tritrichomonas muris (T. muris) infection of mice within an SPF facility. Although T. muris has been considered a commensal organism, we observed instances of asymmetric infection in gene-knockout mice as compared to wild-type mice. We utilized treatment with metronidazole and confirmed successful elimination of T. muris from our SPF colony using extraction of fecal DNA followed by PCR detection. We propose that T. muris testing should be considered for SPF mice, particularly in immunity studies.
]]></description>
<dc:creator>Da Costa, A. S.</dc:creator>
<dc:creator>Graham, T. M.</dc:creator>
<dc:creator>Duncan, J. A.</dc:creator>
<dc:creator>Pillai, S. P. S.</dc:creator>
<dc:creator>Lund, J. M.</dc:creator>
<dc:date>2019-11-01</dc:date>
<dc:identifier>doi:10.1101/827055</dc:identifier>
<dc:title><![CDATA[Detection and treatment strategy for Tritrichomonas muris in the common laboratory mouse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/836296v1?rss=1">
<title>
<![CDATA[
Novel patterns of complex structural variation revealed across thousands of cancer genome graphs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/836296v1?rss=1</link>
<description><![CDATA[
Cancer genomes often harbor hundreds of somatic DNA rearrangement junctions, many of which cannot be easily classified into simple (e.g. deletion, translocation) or complex (e.g. chromothripsis, chromoplexy) structural variant classes. Applying a novel genome graph computational paradigm to analyze the topology of junction copy number (JCN) across 2,833 tumor whole genome sequences (WGS), we introduce three complex rearrangement phenomena: pyrgo, rigma, and tyfonas. Pyrgo are "towers" of low-JCN duplications associated with early replicating regions and superenhancers, and are enriched in breast and ovarian cancers. Rigma comprise "chasms" of low-JCN deletions at late-replicating fragile sites in esophageal and other gastrointestinal (GI) adenocarcinomas. Tyfonas are "typhoons" of high-JCN junctions and fold back inversions that are enriched in acral but not cutaneous melanoma and associated with a previously uncharacterized mutational process of non-APOBEC kataegis. Clustering of tumors according to genome graph-derived features identifies subgroups associated with DNA repair defects and poor prognosis.
]]></description>
<dc:creator>Hadi, K.</dc:creator>
<dc:creator>Yao, X.</dc:creator>
<dc:creator>Behr, J. M.</dc:creator>
<dc:creator>Deshpande, A.</dc:creator>
<dc:creator>Xanthopoulakis, C.</dc:creator>
<dc:creator>Rosiene, J.</dc:creator>
<dc:creator>Darmofal, M.</dc:creator>
<dc:creator>Tian, H.</dc:creator>
<dc:creator>DeRose, J.</dc:creator>
<dc:creator>Mortensen, R.</dc:creator>
<dc:creator>Adney, E.</dc:creator>
<dc:creator>Gajic, Z.</dc:creator>
<dc:creator>Eng, K.</dc:creator>
<dc:creator>Wala, J. A.</dc:creator>
<dc:creator>Wrzeszczynski, K. O.</dc:creator>
<dc:creator>Arora, K.</dc:creator>
<dc:creator>Shah, M.</dc:creator>
<dc:creator>Emde, A.-K.</dc:creator>
<dc:creator>Felice, V.</dc:creator>
<dc:creator>Frank, M. O.</dc:creator>
<dc:creator>Darnell, R.</dc:creator>
<dc:creator>Ghandi, M.</dc:creator>
<dc:creator>Huang, F.</dc:creator>
<dc:creator>Maciejowski, J.</dc:creator>
<dc:creator>De Lange, T.</dc:creator>
<dc:creator>Setton, J.</dc:creator>
<dc:creator>Riaz, N.</dc:creator>
<dc:creator>Reis-Filho, J. S.</dc:creator>
<dc:creator>Powell, S.</dc:creator>
<dc:creator>Knowles, D.</dc:creator>
<dc:creator>Reznik, E.</dc:creator>
<dc:creator>Mishra, B.</dc:creator>
<dc:creator>Beroukhim, R.</dc:creator>
<dc:creator>Zody, M.</dc:creator>
<dc:creator>Robine, N.</dc:creator>
<dc:creator>Oman, K.</dc:creator>
<dc:creator>Sanchez, C. A.</dc:creator>
<dc:creator>Kuhner, M. K.</dc:creator>
<dc:creator>Smith, L. P.</dc:creator>
<dc:creator>Galipeau, P. C.</dc:creator>
<dc:creator>Paulson, T. G.</dc:creator>
<dc:creator>Reid, B. J.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Wilkes, D.</dc:creator>
<dc:creator>Sboner, A.</dc:creator>
<dc:creator>Mosquera, J. M.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2019-11-09</dc:date>
<dc:identifier>doi:10.1101/836296</dc:identifier>
<dc:title><![CDATA[Novel patterns of complex structural variation revealed across thousands of cancer genome graphs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/836668v1?rss=1">
<title>
<![CDATA[
Aurora B-dependent Ndc80 Degradation Regulates Kinetochore Composition in Meiosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/836668v1?rss=1</link>
<description><![CDATA[
The kinetochore complex is a conserved machinery that connects chromosomes to spindle microtubules. During meiosis, the kinetochore is restructured to accommodate a specialized chromosome segregation pattern. In budding yeast, meiotic kinetochore remodeling is mediated by the temporal changes in the abundance of a single subunit called Ndc80. We have previously described the regulatory events that control the timely synthesis of Ndc80. Here, we report that Ndc80 turnover is also tightly regulated in meiosis: Ndc80 degradation is active in meiotic prophase, but not in metaphase I. Ndc80 degradation depends on the ubiquitin ligase APCAma1 and is mediated by the proteasome. Importantly, Aurora B-dependent Ndc80 phosphorylation, a mark that has been previously implicated in correcting erroneous microtubule-kinetochore attachments, is essential for Ndc80 degradation in a microtubule-independent manner. The N-terminus of Ndc80, including a 27-residue sequence and Aurora B phosphorylation sites, is both necessary and sufficient for kinetochore protein degradation. Finally, defects in Ndc80 turnover predispose meiotic cells to chromosome mis-segregation. Our study elucidates the mechanism by which meiotic cells modulate their kinetochore composition through regulated Ndc80 degradation, and demonstrates that Aurora B-dependent regulation of kinetochores extends beyond altering microtubule attachments.
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Liao, A.</dc:creator>
<dc:creator>Powers, E. N.</dc:creator>
<dc:creator>Liao, H.</dc:creator>
<dc:creator>Kohlstaedt, L. A.</dc:creator>
<dc:creator>Evans, R.</dc:creator>
<dc:creator>Holly, R. M.</dc:creator>
<dc:creator>Kim Kim, J.</dc:creator>
<dc:creator>Jovanovic, M.</dc:creator>
<dc:creator>Unal, E.</dc:creator>
<dc:date>2019-11-09</dc:date>
<dc:identifier>doi:10.1101/836668</dc:identifier>
<dc:title><![CDATA[Aurora B-dependent Ndc80 Degradation Regulates Kinetochore Composition in Meiosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/837138v1?rss=1">
<title>
<![CDATA[
Beyond Conventional Models: Recreating the Initiation, Evolution, and Genome of GBM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/837138v1?rss=1</link>
<description><![CDATA[
BackgroundImagining ways to prevent or treat glioblastoma (GBM) have been hindered by a lack of understanding of its pathogenesis. Although PDGF-AA overexpression may be an early event, critical details of the core biology are lacking. Existing PDGF-driven models replicate its microscopic appearance but not the genomic architecture characteristic of the human disease. Here we report a new model of GBM that overcomes this barrier to authenticity.

MethodsUsing a method developed to study neural stem cells, we investigated the effects of PDGF-AA on subventricular zone (SVZ) cells, the putative cell of origin of GBM. We micro-dissected SVZ tissue from p53-null and wild-type adult mice, established primary cultures in media supplemented with PDGF-AA, and assessed cell viability, proliferation, genome stability, and tumour forming potential.

ResultsCounterintuitive to its canonical role as a growth factor, we observed immediate and massive death of SVZ cells in PDGF-AA. Wild-type cells did not survive in PDGF-AA. However, a small fraction of null cells evaded apoptosis, displayed attenuated proliferation, gradually accumulated whole chromosome gains and losses, and, signalled by sudden rapid proliferation and growth factor independence, became tumorigenic in immune-competent syngeneic mice. Transformed cells had an OPC-like profile, were resistant to PDGFR- inhibition, and harboured highly abnormal karyotypes similar to those seen in human GBMs.

ConclusionThis model associates genome instability in SVZ cells with chronic exposure to PDGF-AA; it is the first model to replicate the genomic landscape of GBM and first in which the earliest phases of GBM can be directly observed.

IMPORTANCE OF STUDYWe have developed a mouse model in which the initiation, evolution and genomic landscape of GBM can be thoroughly studied thus paving the way for ideas about how this deadly brain cancer might be prevented, interrupted at an occult stage, or treated with very different therapies.
]]></description>
<dc:creator>Bohm, A.</dc:creator>
<dc:creator>DePetro, J.</dc:creator>
<dc:creator>Binding, C.</dc:creator>
<dc:creator>Gerber, A.</dc:creator>
<dc:creator>Chahley, N.</dc:creator>
<dc:creator>Ware, M.</dc:creator>
<dc:creator>Thomas, K.</dc:creator>
<dc:creator>Bukhari, S.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Chahley, E.</dc:creator>
<dc:creator>Grisdale, C.</dc:creator>
<dc:creator>Berger, N. D.</dc:creator>
<dc:creator>Lawn, S.</dc:creator>
<dc:creator>Yu, Y.</dc:creator>
<dc:creator>Wong, R.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Omairi, H.</dc:creator>
<dc:creator>Mirzaei, R.</dc:creator>
<dc:creator>Maxwell, L.</dc:creator>
<dc:creator>Pedersen, H.</dc:creator>
<dc:creator>Yong, V. W.</dc:creator>
<dc:creator>Weiss, S.</dc:creator>
<dc:creator>Chan, J. A.</dc:creator>
<dc:creator>Cimino, P. J.</dc:creator>
<dc:creator>Kelly, J.</dc:creator>
<dc:creator>Jones, S. J. M.</dc:creator>
<dc:creator>Holland, E.</dc:creator>
<dc:creator>Blough, M. D.</dc:creator>
<dc:creator>Cairncross, G.</dc:creator>
<dc:date>2019-11-09</dc:date>
<dc:identifier>doi:10.1101/837138</dc:identifier>
<dc:title><![CDATA[Beyond Conventional Models: Recreating the Initiation, Evolution, and Genome of GBM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/840348v1?rss=1">
<title>
<![CDATA[
Mediator subunit Med15 dictates the conserved 'fuzzy' binding mechanism of yeast transcription activators Gal4 and Gcn4 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/840348v1?rss=1</link>
<description><![CDATA[
The acidic activation domain (AD) of yeast transcription factor Gal4 plays a dual role in both transcription repression and activation through sequence-dependent binding to Gal80 repressor and sequence-independent binding to Mediator subunit Med15. The activation function of Gal4 arises from two hydrophobic regions within the 40-residue AD. We show by NMR that each AD region binds the Mediator subunit Med15 using a "fuzzy" protein interface. Remarkably, comparison of chemical shift perturbations shows that Gal4 and Gcn4, two ADs of different sequence, interact nearly identically with Med15. The findings that two ADs of different sequence use an identical fuzzy binding mechanism shows a common sequence-independent mechanism for AD-Mediator binding, similar to interactions within a hydrophobic cloud. In contrast, the same region of Gal4 AD interacts with Gal80 via a tight structured complex, implying that the structured binding partner of an intrinsically disordered protein dictates the type of protein interaction.
]]></description>
<dc:creator>Tuttle, L. M.</dc:creator>
<dc:creator>Pacheco, D.</dc:creator>
<dc:creator>Warfield, L.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:creator>Klevit, R. E.</dc:creator>
<dc:date>2019-11-13</dc:date>
<dc:identifier>doi:10.1101/840348</dc:identifier>
<dc:title><![CDATA[Mediator subunit Med15 dictates the conserved 'fuzzy' binding mechanism of yeast transcription activators Gal4 and Gcn4]]></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/10.1101/841395v1?rss=1">
<title>
<![CDATA[
Comparative Genomics and Full-Length TprK Profiling of Treponema pallidum subsp. pallidum Reinfection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/841395v1?rss=1</link>
<description><![CDATA[
Developing a vaccine against Treponema pallidum subspecies pallidum, the causative agent of syphilis, remains a public health priority. Syphilis vaccine design efforts have been complicated by lack of an in vitro T. pallidum culture system, prolific antigenic variation in outer membrane protein TprK, and lack of functional annotation for nearly half of the genes. Understanding the genetic basis of T. pallidum reinfection can provide insights into variation among strains that escape cross-protective immunity. Here, we present comparative genomic sequencing and deep, full-length tprK profiling of two T. pallidum isolates from blood from the same patient that were collected six years apart. Notably, this patient was diagnosed with syphilis four times, with two of these episodes meeting the definition of neurosyphilis, during this interval. Outside of the highly variable tprK gene, we identified 14 coding changes in 13 genes. Nine of these genes putatively localized to the periplasmic or outer membrane spaces, consistent with a potential role in serological immunoevasion. Using a newly developed full-length tprK deep sequencing protocol, we profiled the diversity of this gene that far outpaces the rest of the genome. Intriguingly, we found that the reinfecting isolate demonstrated less diversity across each tprK variable region compared to the isolate from the first infection. Notably, the two isolates did not share any full-length TprK sequences. Our results are consistent with an immunodominant-evasion model in which the diversity of TprK explains the ability of T. pallidum to successfully reinfect individuals, even when they have been infected with the organism multiple times.

Author SummaryThe causative agent of syphilis, Treponema pallidum subspecies pallidum, is capable of repeat infections in people, suggesting that the human immune response does not develop sufficiently broad or long-lasting immunity to cover treponemal diversity. Here, we examined the genomes from two blood-derived isolates of T. pallidum derived 6 years apart from a patient who had syphilis four times during the same period to understand the genetic basis of reinfection. We found a paucity of coding changes across the genome outside of the highly variable tprK gene. Using deep profiling of the full-length tprK gene, we found surprisingly that the two isolates did not share any full-length TprK sequences.
]]></description>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Tantalo, L. C.</dc:creator>
<dc:creator>Lin, M. J.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Huang, M.-L.</dc:creator>
<dc:creator>Marra, C.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:date>2019-11-13</dc:date>
<dc:identifier>doi:10.1101/841395</dc:identifier>
<dc:title><![CDATA[Comparative Genomics and Full-Length TprK Profiling of Treponema pallidum subsp. pallidum Reinfection]]></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/10.1101/843219v1?rss=1">
<title>
<![CDATA[
Distinct roles for the Charcot-Marie-Tooth disease-causing endosomal regulators Mtmr5 and Mtmr13 in axon radial sorting and Schwann cell myelination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/843219v1?rss=1</link>
<description><![CDATA[
The form of Charcot-Marie-Tooth type 4B (CMT4B) disease caused by mutations in myotubularin-related 5 (MTMR5; also called SET Binding Factor 1; SBF1) shows a spectrum of axonal and demyelinating nerve phenotypes. This contrasts with the CMT4B subtypes caused by MTMR2 or MTMR13 (SBF2) mutations, which are characterized by myelin outfoldings and classic demyelination. Thus, it is unclear whether MTMR5 plays an analogous or distinct role from that of its homolog, MTMR13, in the peripheral nervous system (PNS). MTMR5 and MTMR13 are pseudophosphatases predicted to regulate endosomal trafficking by activating Rab GTPases and binding to the phosphoinositide 3-phosphatase MTMR2. In the mouse PNS, Mtmr2 was required to maintain wild type levels of Mtmr5 and Mtmr13, suggesting that these factors function in discrete protein complexes. Genetic elimination of both Mtmr5 and Mtmr13 in mice led to perinatal lethality, indicating that the two proteins have partially redundant functions during embryogenesis. Loss of Mtmr5 in mice did not cause CMT4B-like myelin outfoldings. However, adult Mtmr5-/- mouse nerves contained fewer myelinated axons than control nerves, likely as a result of axon radial sorting defects. Mtmr5 levels were highest during axon radial sorting, whereas Mtmr13 levels rose as myelin formed, and remained high through adulthood. Our findings suggest that Mtmr5 and Mtmr13 ensure proper axon radial sorting and Schwann cell myelination, respectively, perhaps through their direct interactions with Mtmr2. This study enhances our understanding of the non-redundant roles of the endosomal regulators MTMR5 and MTMR13 during normal peripheral nerve development and disease.
]]></description>
<dc:creator>Mammel, A. E.</dc:creator>
<dc:creator>Delgado, K. C.</dc:creator>
<dc:creator>Chin, A. L.</dc:creator>
<dc:creator>Condon, A. F.</dc:creator>
<dc:creator>Hill, J. Q.</dc:creator>
<dc:creator>Aicher, S. A.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Fedorov, L. M.</dc:creator>
<dc:creator>Robinson, F. L.</dc:creator>
<dc:date>2019-11-15</dc:date>
<dc:identifier>doi:10.1101/843219</dc:identifier>
<dc:title><![CDATA[Distinct roles for the Charcot-Marie-Tooth disease-causing endosomal regulators Mtmr5 and Mtmr13 in axon radial sorting and Schwann cell myelination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/844191v1?rss=1">
<title>
<![CDATA[
Enhancers predominantly regulate gene expression in vivo via transcription initiation. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/844191v1?rss=1</link>
<description><![CDATA[
Gene transcription occurs via a cycle of linked events including initiation, promoter proximal pausing and elongation of RNA polymerase II (Pol II). A key question is how do transcriptional enhancers influence these events to control gene expression? Here we have used a new approach to quantify transcriptional initiation and pausing in vivo, while simultaneously identifying transcription start sites (TSSs) and pause-sites (TPSs) from single RNA molecules. When analyzed in parallel with nascent RNA-seq, these data show that differential gene expression is achieved predominantly via changes in transcription initiation rather than Pol II pausing. Using genetically engineered mouse models deleted for specific enhancers we show that these elements control gene expression via Pol II recruitment and/or initiation rather than via promoter proximal pause release. Together, our data show that enhancers, in general, control gene expression predominantly by Pol II recruitment and initiation rather than via pausing.
]]></description>
<dc:creator>Larke, M. S. C.</dc:creator>
<dc:creator>Nojima, T.</dc:creator>
<dc:creator>Telenius, J. M.</dc:creator>
<dc:creator>Sharpe, J. A.</dc:creator>
<dc:creator>Stanley, J. A.</dc:creator>
<dc:creator>Butler, S.</dc:creator>
<dc:creator>Beagrie, R. A.</dc:creator>
<dc:creator>Downes, D. J.</dc:creator>
<dc:creator>Schwessinger, R.</dc:creator>
<dc:creator>Oudelaar, M.</dc:creator>
<dc:creator>Truch, J.</dc:creator>
<dc:creator>Crompton, B.</dc:creator>
<dc:creator>Bender, M. A.</dc:creator>
<dc:creator>Proudfoot, N. J.</dc:creator>
<dc:creator>Higgs, D. R.</dc:creator>
<dc:creator>Hughes, J. R.</dc:creator>
<dc:date>2019-11-16</dc:date>
<dc:identifier>doi:10.1101/844191</dc:identifier>
<dc:title><![CDATA[Enhancers predominantly regulate gene expression in vivo via transcription initiation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/849281v1?rss=1">
<title>
<![CDATA[
Kinome-centric pharmacoproteomics identifies signaling pathways underlying cellular responses to targeted cancer drugs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/849281v1?rss=1</link>
<description><![CDATA[
ABSTRACTKinase-dependent signaling networks are frequently dysregulated in cancer, driving disease progression. While kinase inhibition has become an important therapeutic approach many cancers resist drug treatment. Therefore, we need both reliable biomarkers that predict drug responses and new targets to overcome drug resistance. Determining the kinase(s) that control cancer progression in individual cancers can pose a significant challenge. Genomics has identified important, yet limited numbers of kinase driver mutations. Transcriptomics can quantify aberrant gene expression, but it cannot measure the protein phosphorylation that regulates kinase-dependent signaling network activity. Proteomics measures protein expression and phosphorylation and, therefore, quantifies aberrant signaling network activity directly. We developed a kinome-centric pharmacoproteomics platform to study signaling pathways that determine cancer drug response. Using hepatocellular carcinoma (HCC) as our model, we determined kinome activity with kinobead/LC-MS profiling, and screened 299 kinase inhibitors for growth inhibition. Integrating kinome activity with drug responses, we obtained a comprehensive database of predictive biomarkers, and kinase targets that promote drug sensitivity and resistance. Our dataset specified pathway-based biomarkers for the clinical HCC drugs sorafenib, regorafenib and lenvatinib, and we found these biomarkers enriched in human HCC specimens. Strikingly, our database also revealed signaling pathways that promote HCC cell epithelial-mesenchymal transition (EMT) and drug resistance, and that NUAK1 and NUAK2 regulate these pathways. Inhibition of these kinases reversed the EMT and sensitized HCC cells to kinase inhibition. These results demonstrate that our kinome pharmacoproteomics platform discovers both predictive biomarkers for personalized oncology and novel cancer drug targets.
]]></description>
<dc:creator>Golkowski, M.</dc:creator>
<dc:creator>Lau, H.-T.</dc:creator>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Kenerson, H.</dc:creator>
<dc:creator>Vidadala, V. N.</dc:creator>
<dc:creator>Shoemaker, A.</dc:creator>
<dc:creator>Maly, D. J.</dc:creator>
<dc:creator>Yeung, R. S.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:creator>Ong, S.-E.</dc:creator>
<dc:date>2019-11-20</dc:date>
<dc:identifier>doi:10.1101/849281</dc:identifier>
<dc:title><![CDATA[Kinome-centric pharmacoproteomics identifies signaling pathways underlying cellular responses to targeted cancer drugs]]></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/10.1101/850404v1?rss=1">
<title>
<![CDATA[
alignparse: A Python package for parsing complex features from high-throughput long-read sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/850404v1?rss=1</link>
<description><![CDATA[
Summary & PurposeAdvances in sequencing technology have made it possible to generate large numbers of long, high-accuracy sequencing reads. For instance, the new PacBio Sequel platform can generate hundreds of thousands of high-quality circular consensus sequences in a single run (Rhoads and F. 2015; Hebert et al. 2018). Good programs exist for aligning these reads for genome assembly (Chaisson and Tesler 2012; Li 2018). However, these long reads can also be used for other purposes, such as sequencing PCR amplicons that contain various features of interest. For instance, PacBio circular consensus sequences have been used to identify the mutations in influenza viruses in single cells (Russell et al. 2019), or to link barcodes to gene mutants in deep mutational scanning (Matreyek et al. 2018). For such applications, the alignment of the sequences to the targets may be fairly trivial, but it is not trivial to then parse specific features of interest (such as mutations, unique molecular identifiers, cell barcodes, and flanking sequences) from these alignments.

Here we describe alignparse, a Python package for parsing complex sets of features from long sequences that map to known targets. Specifically, it allows the user to provide complex target sequences in Genbank format that contain an arbitrary number of user-defined sub-sequence features. It then aligns the sequencing reads to these targets and filters alignments based on whether the user-specified features are present with the desired identities (which can be set to different thresholds for different features). Finally, it parses out the sequences, mutations, and/or accuracy of these features as specified by the user. The flexibility of this package therefore fulfills the need for a tool to extract and analyze complex sets of features in large numbers of long sequencing reads.
]]></description>
<dc:creator>Crawford, K. H.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2019-11-21</dc:date>
<dc:identifier>doi:10.1101/850404</dc:identifier>
<dc:title><![CDATA[alignparse: A Python package for parsing complex features from high-throughput long-read sequencing]]></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/10.1101/850479v1?rss=1">
<title>
<![CDATA[
Sort-purification of human CD34+CD90+ cells reduces target cell population and improves lentiviral transduction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/850479v1?rss=1</link>
<description><![CDATA[
ABSTRACTHematopoietic stem cell (HSC) gene therapy has the potential to cure many genetic, malignant and infectious diseases. We have shown in a nonhuman primate (NHP) HSC gene therapy and transplantation model that the CD34+CD90+ cell fraction was exclusively responsible for multilineage engraftment and hematopoietic reconstitution. Here we show the translational potential of this HSC-enriched CD34 subset for lentivirus-mediated gene therapy. Alternative HSC-enrichment strategies include the purification of CD133+ cells or CD38low/- subsets of CD34+ cells from human blood products. We directly compared these strategies to the isolation of CD90+ cells using a GMP-grade flow-sorting protocol with clinical applicability. We show that CD90+ cell selection results in 40-fold fewer target cells in comparison to CD133+ or CD38low/- CD34 subsets without compromising the engraftment potential in vivo. Single cell RNA sequencing confirmed nearly complete depletion of lineage committed progenitor cells in CD90+ fractions compared to alternative selections. Importantly, lentiviral transduction efficiency in purified CD90+ cells resulted in up to 3-fold higher levels of engrafted gene-modified blood cells. These studies should have important implications for the manufacturing of patient-specific HSC gene therapy and genome editing products.
]]></description>
<dc:creator>Radtke, S.</dc:creator>
<dc:creator>Pande, D.</dc:creator>
<dc:creator>Cui, M.</dc:creator>
<dc:creator>Perez, A. M.</dc:creator>
<dc:creator>Chan, Y.-Y.</dc:creator>
<dc:creator>Enstrom, M.</dc:creator>
<dc:creator>Schmuck, S.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:creator>Eunson, T.</dc:creator>
<dc:creator>Adair, J. E.</dc:creator>
<dc:creator>Kiem, H.-P.</dc:creator>
<dc:date>2019-11-26</dc:date>
<dc:identifier>doi:10.1101/850479</dc:identifier>
<dc:title><![CDATA[Sort-purification of human CD34+CD90+ cells reduces target cell population and improves lentiviral transduction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/856476v1?rss=1">
<title>
<![CDATA[
Visual Cell Sorting: A High-throughput, Microscope-based Method to Dissect Cellular Heterogeneity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/856476v1?rss=1</link>
<description><![CDATA[
Microscopy is a powerful tool for characterizing complex cellular phenotypes, but linking these phenotypes to genotype or RNA expression at scale remains challenging. Here, we present Visual Cell Sorting, a method that physically separates hundreds of thousands of live cells based on their visual phenotype. Visual Cell Sorting uses automated imaging and phenotypic analysis to direct selective illumination of Dendra2, a photoconvertible fluorescent protein expressed in live cells; these photoactivated cells are then isolated using fluorescence-activated cell sorting. First, we use Visual Cell Sorting to assess the effect of hundreds of nuclear localization sequence variants in a pooled format, identifying variants that improve nuclear localization and enabling annotation of nuclear localization sequences in thousands of human proteins. Second, we use Visual Cell Sorting to recover cells that retain normal nuclear morphologies after paclitaxel treatment, then derive their single cell transcriptomes to identify multiple pathways associated with paclitaxel resistance in human cancers. Unlike alternative methods, Visual Cell Sorting depends on inexpensive reagents and commercially available hardware. As such, it can be readily deployed to uncover the relationships between visual cellular phenotypes and internal states, including genotypes and gene expression programs.
]]></description>
<dc:creator>Hasle, N.</dc:creator>
<dc:creator>Cooke, A.</dc:creator>
<dc:creator>Srivatsan, S.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Stephany, J. J.</dc:creator>
<dc:creator>Krieger, Z.</dc:creator>
<dc:creator>Jackson, D. L.</dc:creator>
<dc:creator>Tang, W.</dc:creator>
<dc:creator>Monnat, R. J.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Hatch, E. M.</dc:creator>
<dc:creator>Fowler, D.</dc:creator>
<dc:date>2019-11-26</dc:date>
<dc:identifier>doi:10.1101/856476</dc:identifier>
<dc:title><![CDATA[Visual Cell Sorting: A High-throughput, Microscope-based Method to Dissect Cellular Heterogeneity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/863043v1?rss=1">
<title>
<![CDATA[
Identification of HIV-1 Envelope Mutations that Enhance Entry Using Macaque CD4 and CCR5 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/863043v1?rss=1</link>
<description><![CDATA[
Although Rhesus macaques are an important animal model for HIV-1 vaccine development research, most transmitted HIV-1 strains replicate poorly in macaque cells. A major genetic determinant of this species-specific restriction is a non-synonymous mutation in macaque CD4 that results in reduced HIV-1 Envelope (Env)-mediated viral entry compared to human CD4. Recent research efforts employing either laboratory evolution or structure-guided design strategies have uncovered several mutations in Envs gp120 subunit that enhance binding of macaque CD4 by transmitted/founder HIV-1 viruses. In order to identify additional Env mutations that promote infection of macaque cells, we utilized deep mutational scanning to screen thousands of Env point mutants for those that enhance HIV-1 entry via macaque receptors. We identified many uncharacterized amino acid mutations in the N-terminal heptad repeat (NHR) and C-terminal heptad repeat (CHR) regions of gp41 that increased entry into cells bearing macaque receptors by up to 38-fold. Many of these mutations also modestly increased infection of cells bearing human CD4 and CCR5 (up to 13-fold). NHR/CHR mutations identified by deep mutational scanning that enhanced entry also increased sensitivity to neutralizing antibodies targeting the MPER epitope, and to inactivation by cold-incubation, suggesting that they promote sampling of an intermediate trimer conformation between closed and receptor bound states. Identification of this set of mutations can inform future macaque model studies, and also further our understanding of the relationship between Env structure and function.

ImportanceAlthough Rhesus macaques are the favored non-human primate animal model used in HIV-1 research, most circulating HIV-1 strains poorly infect macaque cells. Studies using macaques to model HIV-1 infection often use evolved, or mutant HIV-1 variants that are able to utilize macaque CD4, but these HIV-1 variants poorly model infection by circulating strains. In this work, we sought to identity HIV-1 mutations that would allow entry into macaque cells, but that would maintain critical characteristics of circulating HIV-1 strains. We employed a powerful experimental method to simultaneously assess the effects of thousands of individual HIV-1 mutations on infection of cells bearing macaque receptors. We identified many previously uncharacterized mutations that enhance infection of circulating HIV-1 strains into cells bearing macaque receptors by up to 38-fold. Identification of these mutations may be of use in future macaque model studies.
]]></description>
<dc:creator>Roop, J. I.</dc:creator>
<dc:creator>Cassidy, N. A.</dc:creator>
<dc:creator>Dingens, A. S.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2019-12-03</dc:date>
<dc:identifier>doi:10.1101/863043</dc:identifier>
<dc:title><![CDATA[Identification of HIV-1 Envelope Mutations that Enhance Entry Using Macaque CD4 and CCR5]]></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/10.1101/872028v1?rss=1">
<title>
<![CDATA[
eVIP2: Expression-based variant impact phenotyping to predict the function of gene variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/872028v1?rss=1</link>
<description><![CDATA[
While advancements in genome sequencing have identified millions of somatic mutations in cancer, their functional impact is poorly understood. We previously developed the expression-based variant impact phenotyping (eVIP) method to use gene expression data to characterize the function of gene variants. The eVIP method uses a decision tree-based algorithm to predict the functional impact of somatic variants by comparing gene expression signatures induced by introduction of wild-type versus mutant cDNAs in cell lines. The method distinguishes between variants that are gain-of-function, loss-of-function, change-of-function, or neutral. We present eVIP2, software that allows for pathway analysis (eVIP Pathways) and usage with RNA-seq data. To demonstrate the eVIP2 software and approach, we characterized two recurrent frameshift variants in RNF43, a negative regulator of Wnt signaling, frequently mutated in colorectal, gastric and endometrial cancer. RNF43 WT, RNF43 R117fs, RNF43 G659fs, or GFP control cDNA were overexpressed in HEK293T cells. Analysis with eVIP2 predicted that the frameshift at position 117 was a loss-of-function mutation, as expected. The second frameshift at position 659, was, surprisingly, predicted to be a gain-of-function mutation. Additional eVIP Pathways analysis of RNF43 G659fs predicted 10 pathways to be significantly altered, including TNF alpha via NFKB signaling, KRAS signaling, and hypoxia. To validate these predictions, we performed reporter assays and found that all eVIP2 impactful pathways tested in the assay were activated by expression of RNF43 G659fs, but not by expression of RNF43 WT, supporting that RNF43 G659fs is a gain-of-function mutation and its effect on the identified pathways. The eVIP2 method is an important step towards overcoming the current challenge of variant interpretation in the implementation of precision medicine. eVIP2 is available at https://github.com/BrooksLabUCSC/eVIP2.
]]></description>
<dc:creator>Thornton, A. M.</dc:creator>
<dc:creator>Fang, L.</dc:creator>
<dc:creator>O'Brian, C.</dc:creator>
<dc:creator>Berger, A. H.</dc:creator>
<dc:creator>Giannakis, M.</dc:creator>
<dc:creator>Brooks, A. N.</dc:creator>
<dc:date>2019-12-11</dc:date>
<dc:identifier>doi:10.1101/872028</dc:identifier>
<dc:title><![CDATA[eVIP2: Expression-based variant impact phenotyping to predict the function of gene variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.30.542944v1?rss=1">
<title>
<![CDATA[
A high-content siRNA screen reveals new regulators of nuclear membrane stability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.30.542944v1?rss=1</link>
<description><![CDATA[
Nuclear membrane rupture is a physiological response to multiple in vivo processes, such as cell migration, that can cause extensive genome instability and upregulate invasive and inflammatory pathways. However, the underlying molecular mechanisms of rupture are unclear and few regulators have been identified. In this study, we developed a reporter that is size excluded from re-compartmentalization following nuclear rupture events. This allows for robust detection of factors influencing nuclear integrity in fixed cells. We combined this with an automated image analysis pipeline in a high-content siRNA screen to identify new proteins that both increase and decrease nuclear rupture frequency in cancer cells. Pathway analysis identified an enrichment of nuclear membrane and ER factors in our hits and we demonstrate that one of these, the protein phosphatase CTDNEP1, is required for nuclear stability. Further analysis of known rupture contributors, including a newly developed automated quantitative analysis of nuclear lamina gaps, strongly suggests that CTDNEP1 acts in a new pathway. Our findings provide new insights into the molecular mechanism of nuclear rupture and define a highly adaptable program for rupture analysis that removes a substantial barrier to new discoveries in the field.
]]></description>
<dc:creator>Gunn, A. L.</dc:creator>
<dc:creator>Yashchenko, A. I.</dc:creator>
<dc:creator>Hatch, E. L.</dc:creator>
<dc:date>2023-05-31</dc:date>
<dc:identifier>doi:10.1101/2023.05.30.542944</dc:identifier>
<dc:title><![CDATA[A high-content siRNA screen reveals new regulators of nuclear membrane stability]]></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/10.1101/2023.06.01.543064v1?rss=1">
<title>
<![CDATA[
Phylodynamics Uncovers the Transmission of Antibiotic-Resistant Escherichia coli between Canines and Humans in an Urban Environment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.01.543064v1?rss=1</link>
<description><![CDATA[
The role of canines in transmitting antibiotic resistant bacteria to humans in the urban environment is poorly understood. To elucidate this role, we utilized genomic sequencing and phylogenetics to characterize the burden and transmission dynamics of antibiotic resistant Escherichia coli (ABR-Ec) cultured from canine and human feces present on urban sidewalks in San Francisco, California. We collected a total of fifty-nine ABR-Ec from human (n=12) and canine (n=47) fecal samples from the Tenderloin and South of Market (SoMa) neighborhoods of San Francisco. We then analyzed phenotypic and genotypic antibiotic resistance (ABR) of the isolates, as well as clonal relationships based on cgMLST and single nucleotide polymorphisms (SNPs) of the core genomes. Using Bayesian inference, we reconstructed the transmission dynamics between humans and canines from multiple local outbreak clusters using the marginal structured coalescent approximation (MASCOT). Overall, we found human and canine samples to carry similar amounts and profiles of ABR genes. Our results provide evidence for multiple transmission events of ABR-Ec between humans and canines. In particular, we found one instance of likely transmission from canines to humans as well as an additional local outbreak cluster consisting of one canine and one human sample. Based on this analysis, it appears that canine feces act as an important reservoir of clinically relevant ABR-Ec within the urban environment. Our findings support that public health measures should continue to emphasize proper canine feces disposal practices, access to public toilets and sidewalk and street cleaning. Importance: Antibiotic resistance in E. coli is a growing public health concern with global attributable deaths projected to reach millions annually. Current research has focused heavily on clinical routes of antibiotic resistance transmission to design interventions while the role of alternative reservoirs such as domesticated animals remain less well understood. Our results suggest canines are part of the transmission network that disseminates high-risk multidrug resistance in E. coli within the urban San Francisco community. As such, this study highlights the need to consider canines, and potentially domesticated animals more broadly, when designing interventions to reduce the prevalence of antibiotic resistance in the community. Additionally, it showcases the utility of genomic epidemiology to reconstruct the pathways by which antimicrobial resistance spreads.
]]></description>
<dc:creator>Walas, N.</dc:creator>
<dc:creator>Müller, N. F.</dc:creator>
<dc:creator>Parker, E.</dc:creator>
<dc:creator>Henderson, A.</dc:creator>
<dc:creator>Capone, D.</dc:creator>
<dc:creator>Brown, J.</dc:creator>
<dc:creator>Barker, T.</dc:creator>
<dc:creator>Graham, J. P.</dc:creator>
<dc:date>2023-06-03</dc:date>
<dc:identifier>doi:10.1101/2023.06.01.543064</dc:identifier>
<dc:title><![CDATA[Phylodynamics Uncovers the Transmission of Antibiotic-Resistant Escherichia coli between Canines and Humans in an Urban Environment]]></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/10.1101/2023.05.31.543119v1?rss=1">
<title>
<![CDATA[
Atg8-LC3 controls systemic nutrient surplus signaling from flies to humans. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.31.543119v1?rss=1</link>
<description><![CDATA[
Organisms experience constant nutritional flux, and homeostatic mechanisms evolved to operate at the nexus of extreme nutritional states - scarcity and surplus. Thus, we surmised that decoding bidirectional molecular switches that operate at the interface of scarcity response and surplus signaling will enable the development of strategies to treat disorders that arise from nutrient imbalance states. Adipocytes secrete leptin, an interleukin protein, which signals nutrient surplus to the central brain to regulate feeding and energy expenditure. We report that Atg8-LC3-family proteins, best known for their role in autophagy, are required for leptin secretion in Drosophila and human adipocytes. Atg8-LC3 genetic knockdown and point mutations to the Atg8-LC3 interaction motif (AIM/LIR) of leptin, and its functional ortholog in Drosophila, Upd2, disrupt adipokine secretion and increase adipokine retention in human and fly cells. At an organismal level, Atg8-driven Upd2 retention increases organismal resilience to nutrient extremes by rewiring the transcriptome, organismal feeding behavior, and hunger response. Comparative proteomic analyses reveal that LC3 directs leptin to an exosome secretory pathway. We use genetic knockdown in primary human adipocytes to establish that LC3 is required for leptin secretion in a physiologically relevant mammalian system. Hence, we uncover a previously unknown and evolutionarily conserved role for Atg8-LC3 in promoting adipocyte-brain nutrient surplus signaling. We propose that Atg8-LC3s bidirectional role in nutrient sensing-conveying nutrient surplus and responding to nutrient deprivation-enables organisms to manage nutrient flux effectively.
]]></description>
<dc:creator>Madan, A.</dc:creator>
<dc:creator>Kelly, K. P.</dc:creator>
<dc:creator>Sullivan, C. E.</dc:creator>
<dc:creator>Poling, M. E.</dc:creator>
<dc:creator>Brent, A. E.</dc:creator>
<dc:creator>Alassaf, M.</dc:creator>
<dc:creator>Dubrulle, J.</dc:creator>
<dc:creator>Rajan, A.</dc:creator>
<dc:date>2023-06-04</dc:date>
<dc:identifier>doi:10.1101/2023.05.31.543119</dc:identifier>
<dc:title><![CDATA[Atg8-LC3 controls systemic nutrient surplus signaling from flies to humans.]]></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/10.1101/2023.06.02.543290v1?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/10.1101/2023.06.01.543314v1?rss=1">
<title>
<![CDATA[
Differentiation latency and dormancy signatures define fetal liver HSCs at single cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.01.543314v1?rss=1</link>
<description><![CDATA[
Decoding the gene regulatory mechanisms and signaling interactions that orchestrate the self-renewal of hematopoietic stem cells (HSCs) during their expansion in the fetal liver (FL) could unlock novel therapeutic strategies to expand transplantable HSCs, a long-standing challenge. Here, to explore intrinsic and extrinsic regulation of FL-HSC self-renewal at the single cell level, we engineered a culture platform designed to recapitulate the FL endothelial niche, which supports the ex vivo amplification of serially engraftable HSCs. Leveraging this platform in combination with single cell index flow cytometry, live imaging, serial transplantation assays, and single cell RNA-sequencing, we uncovered previously unrecognized heterogeneity within immunophenotypically defined FL-HSCs. Specifically, we demonstrated that differentiation latency, symmetric cell divisions, and transcriptional signatures of biosynthetic dormancy and lipid metabolism are distinguishing properties of rare FL-HSCs capable of serial, long-term multilineage hematopoietic reconstitution. Our findings support a paradigm in which intrinsic programs and extrinsic signals combinatorially facilitate the symmetric self-renewal and expansion of nascent HSCs in the FL niche while delaying their active participation in hematopoiesis. Additionally, our study provides a valuable resource for future investigations into the intrinsic and niche-derived signaling pathways that govern FL-HSC self-renewal.
]]></description>
<dc:creator>Ishida, T.</dc:creator>
<dc:creator>Heck, A. M.</dc:creator>
<dc:creator>Varnum-Finney, B.</dc:creator>
<dc:creator>Dozono, S.</dc:creator>
<dc:creator>Nourigat-McKay, C.</dc:creator>
<dc:creator>Kraskouskas, K.</dc:creator>
<dc:creator>Wellington, R.</dc:creator>
<dc:creator>Waltner, O.</dc:creator>
<dc:creator>Root, C.</dc:creator>
<dc:creator>Jackson, D. L.</dc:creator>
<dc:creator>Delaney, C.</dc:creator>
<dc:creator>Rafii, S.</dc:creator>
<dc:creator>Bernstein, I. D.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Hadland, B.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.01.543314</dc:identifier>
<dc:title><![CDATA[Differentiation latency and dormancy signatures define fetal liver HSCs at single cell resolution]]></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/10.1101/2023.06.03.543584v1?rss=1">
<title>
<![CDATA[
A Quantitative Analysis of Locomotor Patterns in Developing C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.03.543584v1?rss=1</link>
<description><![CDATA[
Adult animals display robust locomotion, yet the timeline and mechanisms of how juvenile animals acquire coordinated movements and how these movements evolve during development are not well understood. Recent advances in quantitative behavioral analyses have paved the way for investigating complex natural behaviors like locomotion. In this study, we tracked the swimming and crawling behaviors of the nematode Caenorhabditis elegans from postembryonic development through to adulthood. Our principal component analyses revealed that adult C. elegans swimming is low dimensional, suggesting that a small number of distinct postures, or eigenworms, account for most of the variance in the body shapes that constitute swimming behavior. Additionally, we found that crawling behavior in adult C. elegans is similarly low dimensional, corroborating previous studies. However, our analysis revealed that swimming and crawling are distinct gaits in adult animals, clearly distinguishable within the eigenworm space. Remarkably, young L1 larvae are capable of producing the postural shapes for swimming and crawling seen in adults, despite frequent instances of uncoordinated body movements. In contrast, late L1 larvae exhibit robust coordination of locomotion, while many neurons crucial for adult locomotion are still under development. In conclusion, this study establishes a comprehensive quantitative behavioral framework for understanding the neural basis of locomotor development, including distinct gaits such as swimming and crawling in C. elegans.
]]></description>
<dc:creator>Hassinan, C. W.</dc:creator>
<dc:creator>Sterrett, S. C.</dc:creator>
<dc:creator>Summy, B.</dc:creator>
<dc:creator>Khera, A.</dc:creator>
<dc:creator>Wang, A.</dc:creator>
<dc:creator>Bai, J.</dc:creator>
<dc:date>2023-06-06</dc:date>
<dc:identifier>doi:10.1101/2023.06.03.543584</dc:identifier>
<dc:title><![CDATA[A Quantitative Analysis of Locomotor Patterns in Developing C. elegans]]></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/10.1101/2023.06.10.544477v1?rss=1">
<title>
<![CDATA[
Ablation of Max expression induces meiotic onset in sexually undifferentiated germ cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.10.544477v1?rss=1</link>
<description><![CDATA[
Meiosis is a specialized type of cell division that occurs only in germ cells physiologically. We have previously demonstrated that MYC-associated factor X (MAX) is involved in blocking ectopic and precocious meiotic onsets in embryonic and germline stem cells, respectively, as a central component of the PRC1 subtype PRC1.6. In this study, we investigated the role of the Max gene in germ cells in vivo. Our data revealed that mitotically active germ cell-specific disruption of Max was coupled to meiotic onset in male and female germ cells. However, such Max-null germ cells did not undergo meiotic processes progressively, but were stalled at its relatively early stages and eventually eliminated by apoptosis. Our data also revealed that Max, which is generally known as an obligate heterodimerization partner for MYC and MXD transcription factors, showed high expression in sexually undifferentiated male and female germ cells, but female germ cells exhibited an abrupt decline in its expression at the timing of or immediately prior to physiological meiotic onset. Moreover, computational analyses identified the regulatory region that supported high levels of Max expression in sexually undifferentiated germ cells.
]]></description>
<dc:creator>Suzuki, A.</dc:creator>
<dc:creator>Uranishi, K.</dc:creator>
<dc:creator>Nishimoto, M.</dc:creator>
<dc:creator>Mizuno, Y.</dc:creator>
<dc:creator>Mizuno, S.</dc:creator>
<dc:creator>Takahashi, S.</dc:creator>
<dc:creator>Eisenman, R. N.</dc:creator>
<dc:creator>Okuda, A.</dc:creator>
<dc:date>2023-06-11</dc:date>
<dc:identifier>doi:10.1101/2023.06.10.544477</dc:identifier>
<dc:title><![CDATA[Ablation of Max expression induces meiotic onset in sexually undifferentiated germ cells]]></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/10.1101/2023.06.12.544624v1?rss=1">
<title>
<![CDATA[
De novo design of highly selective miniprotein inhibitors of integrins αvβ6 and αvβ8 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.12.544624v1?rss=1</link>
<description><![CDATA[
The RGD (Arg-Gly-Asp)-binding integrins v{beta}6 and v{beta}8 are clinically validated cancer and fibrosis targets of considerable therapeutic importance. Compounds that can discriminate between the two closely related integrin proteins and other RGD integrins, stabilize specific conformational states, and have sufficient stability enabling tissue restricted administration could have considerable therapeutic utility. Existing small molecules and antibody inhibitors do not have all of these properties, and hence there is a need for new approaches. Here we describe a method for computationally designing hyperstable RGD-containing miniproteins that are highly selective for a single RGD integrin heterodimer and conformational state, and use this strategy to design inhibitors of v{beta}6 and v{beta}8 with high selectivity. The v{beta}6 and v{beta}8 inhibitors have picomolar affinities for their targets, and >1000-fold selectivity over other RGD integrins. CryoEM structures are within 0.6-0.7[A] root-mean-square deviation (RMSD) to the computational design models; the designed v{beta}6 inhibitor and native ligand stabilize the open conformation in contrast to the therapeutic anti-v{beta}6 antibody BG00011 that stabilizes the bent-closed conformation and caused on-target toxicity in patients with lung fibrosis, and the v{beta}8 inhibitor maintains the constitutively fixed extended-closed v{beta}8 conformation. In a mouse model of bleomycin-induced lung fibrosis, the v{beta}6 inhibitor potently reduced fibrotic burden and improved overall lung mechanics when delivered via oropharyngeal administration mimicking inhalation, demonstrating the therapeutic potential of de novo designed integrin binding proteins with high selectivity.
]]></description>
<dc:creator>Roy, A.</dc:creator>
<dc:creator>Shi, L.</dc:creator>
<dc:creator>Chang, A.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Fernandez, A.</dc:creator>
<dc:creator>Kraft, J. C.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Le, V. Q.</dc:creator>
<dc:creator>Winegar, R. V.</dc:creator>
<dc:creator>Cherf, G. M.</dc:creator>
<dc:creator>Slocum, D.</dc:creator>
<dc:creator>Poulson, P. D.</dc:creator>
<dc:creator>casper, G. E.</dc:creator>
<dc:creator>Vallecillo-Zuniga, M. L.</dc:creator>
<dc:creator>Valdoz, J. C.</dc:creator>
<dc:creator>Miranda, M. C.</dc:creator>
<dc:creator>Bai, H.</dc:creator>
<dc:creator>Kipnis, Y.</dc:creator>
<dc:creator>Olshefsky, A.</dc:creator>
<dc:creator>Priya, T.</dc:creator>
<dc:creator>Carter, L.</dc:creator>
<dc:creator>Ravichandran, R.</dc:creator>
<dc:creator>Chow, C.</dc:creator>
<dc:creator>Johnson, M. R.</dc:creator>
<dc:creator>Chang, S.</dc:creator>
<dc:creator>Roy, A.</dc:creator>
<dc:creator>Overed-Sayer, C.</dc:creator>
<dc:creator>Finch, D. K.</dc:creator>
<dc:creator>Lowe, D.</dc:creator>
<dc:creator>Bera, A. K.</dc:creator>
<dc:creator>Matute-Bello, G.</dc:creator>
<dc:creator>Birkland, T. P.</dc:creator>
<dc:creator>DiMaio, F.</dc:creator>
<dc:creator>Raghu, G.</dc:creator>
<dc:creator>Cochran, J. R.</dc:creator>
<dc:creator>Stewart, L.</dc:creator>
<dc:creator>Campbell, M. G.</dc:creator>
<dc:creator>Van Ry, P. M.</dc:creator>
<dc:creator>Springer, T. A.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2023-06-12</dc:date>
<dc:identifier>doi:10.1101/2023.06.12.544624</dc:identifier>
<dc:title><![CDATA[De novo design of highly selective miniprotein inhibitors of integrins αvβ6 and αvβ8]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.16.545380v1?rss=1">
<title>
<![CDATA[
Fusobacterium sphaericum sp. nov., isolated from a human colon tumor, is prevalent in various human body sites and induces IL-8 secretion from colorectal cancer cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.16.545380v1?rss=1</link>
<description><![CDATA[
Cancerous tissue is a largely unexplored microbial niche that provides a unique environment for the colonization and growth of specific bacterial communities, and with it, the opportunity to identify novel bacterial species. Here, we report distinct features of a novel Fusobacterium species, F. sphaericum sp. nov. (Fs), isolated from primary colon adenocarcinoma tissue. We acquire the complete closed genome and associated methylome of this organism and phylogenetically confirm its classification into the Fusobacterium genus, with F. perfoetens as its closest neighbor. Fs is phenotypically and genetically distinct, with morphological analysis revealing its coccoid shape, that while similar to F. perfoetens is rare for most Fusobacterium members. Fs displays a metabolic profile and antibiotic resistance repertoire consistent with other Fusobacterium species. In vitro, Fs has adherent and immunomodulatory capabilities, as it intimately associates with human colon cancer epithelial cells and promotes IL-8 secretion. Analysis of the prevalence and abundance of Fs in >20,000 human metagenomic samples shows that it is a low-prevalence member within human stool with variable relative abundance, found in both healthy controls and patients with colorectal cancer (CRC). Our study sheds light on a novel bacterial species isolated directly from the human CRC tumor niche, and given its interaction with cancer epithelial cells suggests that its role in human health and disease warrants further investigation.
]]></description>
<dc:creator>Zepeda-Rivera, M. A.</dc:creator>
<dc:creator>Eisele, Y.</dc:creator>
<dc:creator>Baryiames, A.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>LaCourse, K. D.</dc:creator>
<dc:creator>Jones, D. S.</dc:creator>
<dc:creator>Hauner, H.</dc:creator>
<dc:creator>Dewhirst, F. E.</dc:creator>
<dc:creator>Minot, S. S.</dc:creator>
<dc:creator>Johnston, C. D.</dc:creator>
<dc:creator>Bullman, S.</dc:creator>
<dc:date>2023-06-16</dc:date>
<dc:identifier>doi:10.1101/2023.06.16.545380</dc:identifier>
<dc:title><![CDATA[Fusobacterium sphaericum sp. nov., isolated from a human colon tumor, is prevalent in various human body sites and induces IL-8 secretion from colorectal cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.16.545341v1?rss=1">
<title>
<![CDATA[
Design of four component T=4 tetrahedral, octahedral, and icosahedral protein nanocages through programmed symmetry breaking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.16.545341v1?rss=1</link>
<description><![CDATA[
Four, eight or twenty C3 symmetric protein trimers can be arranged with tetrahedral (T-sym), octahedral (O-sym) or icosahedral (I-sym) point group symmetry to generate closed cage-like structures1,2. Generating more complex closed structures requires breaking perfect point group symmetry. Viruses do this in the icosahedral case using quasi-symmetry or pseudo-symmetry to access higher triangulation number architectures3-9, but nature appears not to have explored higher triangulation number tetrahedral or octahedral symmetries. Here, we describe a general design strategy for building T = 4 architectures starting from simpler T = 1 structures through pseudo-symmetrization of trimeric building blocks. Electron microscopy confirms the structures of T = 4 cages with 48 (T-sym), 96 (O-sym), and 240 (I-sym) subunits, each with four distinct chains and six different protein-protein interfaces, and diameters of 33nm, 43nm, and 75nm, respectively. Higher triangulation number viruses possess very sophisticated functionalities; our general route to higher triangulation number nanocages should similarly enable a next generation of multiple antigen displaying vaccine candidates10,11 and targeted delivery vehicles12,13.
]]></description>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Kibler, R. D.</dc:creator>
<dc:creator>Hsia, Y.</dc:creator>
<dc:creator>Borst, A. J.</dc:creator>
<dc:creator>Philomin, A.</dc:creator>
<dc:creator>Kennedy, M. A.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2023-06-17</dc:date>
<dc:identifier>doi:10.1101/2023.06.16.545341</dc:identifier>
<dc:title><![CDATA[Design of four component T=4 tetrahedral, octahedral, and icosahedral protein nanocages through programmed symmetry breaking]]></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/10.1101/2023.06.23.544827v1?rss=1">
<title>
<![CDATA[
MOCHA: Advanced statistical modeling of scATAC-seq data enables functional genomic inference in large human disease cohorts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.23.544827v1?rss=1</link>
<description><![CDATA[
Single-cell assay for transposase-accessible chromatin using sequencing (scATAC-seq) has been increasingly used to study gene regulation. However, major analytical gaps limit its utility in studying gene regulatory programs in complex diseases. We developed MOCHA (Model-based single cell Open CHromatin Analysis) with major advances over existing analysis tools, including: 1) improved identification of sample-specific open chromatin, 2) proper handling of technical drop-out with zero-inflated methods, 3) mitigation of false positives in single cell analysis, 4) identification of alternative transcription-starting-site regulation, and 5) transcription factor-gene network construction from longitudinal scATAC-seq data. These advances provide a robust framework to study gene regulatory programs in human disease. We benchmarked MOCHA with four state-of-the-art tools to demonstrate its advances. We also constructed cross-sectional and longitudinal gene regulatory networks, identifying potential mechanisms of COVID-19 response. MOCHA provides researchers with a robust analytical tool for functional genomic inference from scATAC-seq data.
]]></description>
<dc:creator>Rachid Zaim, S.</dc:creator>
<dc:creator>Pebworth, M.-P.</dc:creator>
<dc:creator>McGrath, I.</dc:creator>
<dc:creator>Okada, L.</dc:creator>
<dc:creator>Weiss, M.</dc:creator>
<dc:creator>Reading, J.</dc:creator>
<dc:creator>Czartoski, J. L.</dc:creator>
<dc:creator>Torgerson, T.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Bumol, T. F.</dc:creator>
<dc:creator>Skene, P. J.</dc:creator>
<dc:creator>Li, X.-j.</dc:creator>
<dc:date>2023-06-24</dc:date>
<dc:identifier>doi:10.1101/2023.06.23.544827</dc:identifier>
<dc:title><![CDATA[MOCHA: Advanced statistical modeling of scATAC-seq data enables functional genomic inference in large human disease cohorts]]></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/10.1101/2023.06.27.546775v1?rss=1">
<title>
<![CDATA[
An engineered biosensor enables dynamic aspartate measurements in living cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.27.546775v1?rss=1</link>
<description><![CDATA[
Intracellular levels of the amino acid aspartate are responsive to changes in metabolism in mammalian cells and can correspondingly alter cell function, highlighting the need for robust tools to measure aspartate abundance. However, comprehensive understanding of aspartate metabolism has been limited by the throughput, cost, and static nature of the mass spectrometry based measurements that are typically employed to measure aspartate levels. To address these issues, we have developed a GFP-based sensor of aspartate (jAspSnFR3), where the fluorescence intensity corresponds to aspartate concentration. As a purified protein, the sensor has a 20-fold increase in fluorescence upon aspartate saturation, with dose dependent fluorescence changes covering a physiologically relevant aspartate concentration range and no significant off target binding. Expressed in mammalian cell lines, sensor intensity correlated with aspartate levels measured by mass spectrometry and could resolve temporal changes in intracellular aspartate from genetic, pharmacological, and nutritional manipulations. These data demonstrate the utility of jAspSnFR3 and highlight the opportunities it provides for temporally resolved and high throughput applications of variables that affect aspartate levels.
]]></description>
<dc:creator>Davidsen, K.</dc:creator>
<dc:creator>Marvin, J. S.</dc:creator>
<dc:creator>Aggarwal, A.</dc:creator>
<dc:creator>Brown, T. A.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:date>2023-06-27</dc:date>
<dc:identifier>doi:10.1101/2023.06.27.546775</dc:identifier>
<dc:title><![CDATA[An engineered biosensor enables dynamic aspartate measurements in living cells]]></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/10.1101/2023.06.29.547092v1?rss=1">
<title>
<![CDATA[
Mechanical coupling coordinates microtubule growth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.29.547092v1?rss=1</link>
<description><![CDATA[
During mitosis, kinetochore-attached microtubules form bundles (k-fibers) in which many filaments grow and shorten in near-perfect unison to align and segregate each chromosome. However, individual microtubules grow at intrinsically variable rates, which must be tightly regulated for a k-fiber to behave as a single unit. This exquisite coordination might be achieved biochemically, via selective binding of polymerases and depolymerases, or mechanically, because k-fiber microtubules are coupled through a shared load that influences their growth. Here, we use a novel dual laser trap assay to show that microtubule pairs growing in vitro are coordinated by mechanical coupling. Kinetic analyses show that microtubule growth is interrupted by stochastic, force-dependent pauses and indicate persistent heterogeneity in growth speed during non-pauses. A simple model incorporating both force-dependent pausing and persistent growth speed heterogeneity explains the measured coordination of microtubule pairs without any free fit parameters. Our findings illustrate how microtubule growth may be synchronized during mitosis and provide a basis for modeling k-fiber bundles with three or more microtubules, as found in many eukaryotes.
]]></description>
<dc:creator>Leeds, B. K.</dc:creator>
<dc:creator>Kostello, K. F.</dc:creator>
<dc:creator>Liu, Y. Y.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:creator>Asbury, C. L.</dc:creator>
<dc:date>2023-06-30</dc:date>
<dc:identifier>doi:10.1101/2023.06.29.547092</dc:identifier>
<dc:title><![CDATA[Mechanical coupling coordinates microtubule growth]]></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/10.1101/2023.07.01.547355v1?rss=1">
<title>
<![CDATA[
A Compound that Inhibits Glycolysis in Prostate Cancer Controls Growth of Advanced Prostate Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.01.547355v1?rss=1</link>
<description><![CDATA[
PurposeMetastatic castration-resistant prostate cancer remains incurable regardless of recent therapeutic advances. Prostate cancer tumors display highly glycolytic phenotypes as the cancer progresses. Non-specific inhibitors of glycolysis have not been utilized successfully for chemotherapy, because of their penchant to cause systemic toxicity. This study reports the preclinical activity, safety, and pharmacokinetics of a novel small molecule preclinical candidate, BKIDC-1553, with antiglycolytic activity.

Experimental designWe tested a large battery of prostate cancer cell lines for inhibition of cell proliferation, in vitro. Cell cycle, metabolic and enzymatic assays were used to demonstrate their mechanism of action. A human PDX model implanted in mice and a human organoid were studied for sensitivity to our BKIDC preclinical candidate. A battery of pharmacokinetic experiments, absorption, distribution, metabolism, and excretion experiments, and in vitro and in vivo toxicology experiments were carried out to assess readiness for clinical trials.

ResultsWe demonstrate a new class of small molecule inhibitors where antiglycolytic activity in prostate cancer cell lines is mediated through inhibition of hexokinase 2. These compounds display selective growth inhibition across multiple prostate cancer models. We describe a lead BKIDC-1553 that demonstrates promising activity in a preclinical xenograft model of advanced prostate cancer, equivalent to that of enzalutamide. BKIDC-1553 demonstrates safety and pharmacologic properties consistent with a compound that can be taken into human studies with expectations of a good safety margin and predicted dosing for efficacy.

ConclusionThis work supports testing BKIDC-1553 and its derivatives in clinical trials for patients with advanced prostate cancer.
]]></description>
<dc:creator>Uo, T.</dc:creator>
<dc:creator>Ojo, K. K.</dc:creator>
<dc:creator>Sprenger, C. C.</dc:creator>
<dc:creator>Epilepsia, K. S.</dc:creator>
<dc:creator>Perera, B. G. K.</dc:creator>
<dc:creator>Damodarasamy, M.</dc:creator>
<dc:creator>Sun, S.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Hogan, H. H.</dc:creator>
<dc:creator>Hulverson, M. A.</dc:creator>
<dc:creator>Choi, R.</dc:creator>
<dc:creator>Whitman, G. R.</dc:creator>
<dc:creator>Barrett, L. K.</dc:creator>
<dc:creator>Michaels, S. A.</dc:creator>
<dc:creator>Xu, L. H.</dc:creator>
<dc:creator>Pang, H. J.</dc:creator>
<dc:creator>Nguyen, M. M.</dc:creator>
<dc:creator>Vigil, A.-L. B. G.</dc:creator>
<dc:creator>Kamat, V.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:creator>Sweet, I. R.</dc:creator>
<dc:creator>Vidadala, R.</dc:creator>
<dc:creator>Maly, D. J.</dc:creator>
<dc:creator>Van Voorhis, W. C.</dc:creator>
<dc:creator>Plymate, S. R.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.01.547355</dc:identifier>
<dc:title><![CDATA[A Compound that Inhibits Glycolysis in Prostate Cancer Controls Growth of Advanced Prostate Cancer]]></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/10.1101/2023.07.02.547443v1?rss=1">
<title>
<![CDATA[
APOBEC3 activity promotes the survival and evolution of drug-tolerant persister cells during acquired resistance to EGFR inhibitors in lung cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.02.547443v1?rss=1</link>
<description><![CDATA[
APOBEC mutagenesis is one of the most common endogenous sources of mutations in human cancer and is a major source of genetic intratumor heterogeneity. High levels of APOBEC mutagenesis are associated with poor prognosis and aggressive disease across diverse cancers, but the mechanistic and functional impacts of APOBEC mutagenesis on tumor evolution and therapy resistance remain relatively unexplored. To address this, we investigated the contribution of APOBEC mutagenesis to acquired therapy resistance in a model of EGFR-mutant non-small cell lung cancer. We find that inhibition of EGFR in lung cancer cells leads to a rapid and pronounced induction of APOBEC3 expression and activity. Functionally, APOBEC expression promotes the survival of drug-tolerant persister cells (DTPs) following EGFR inhibition. Constitutive expression of APOBEC3B alters the evolutionary trajectory of acquired resistance to the EGFR inhibitor gefitinib, making it more likely that resistance arises through de novo acquisition of the T790M gatekeeper mutation and squamous transdifferentiation during the DTP state. APOBEC3B expression is associated with increased expression of the squamous cell transcription factor {Delta}Np63 and squamous cell transdifferentiation in gefitinib-resistant cells. Knockout of p63 in gefitinib-resistant cells reduces the expression of the {Delta}Np63 target genes IL1/{beta} and sensitizes these cells to the third-generation EGFR inhibitor osimertinib. These results suggest that APOBEC activity promotes acquired resistance by facilitating evolution and transdifferentiation in DTPs, and suggest that approaches to target {Delta}Np63 in gefitinib-resistant lung cancers may have therapeutic benefit.
]]></description>
<dc:creator>Garcia, N. M. G.</dc:creator>
<dc:creator>Becerra, J. N.</dc:creator>
<dc:creator>McKinney, B. J.</dc:creator>
<dc:creator>DiMarco, A. V.</dc:creator>
<dc:creator>Wu, F.</dc:creator>
<dc:creator>Fitzgibbon, M.</dc:creator>
<dc:creator>Alvarez, J. V.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.02.547443</dc:identifier>
<dc:title><![CDATA[APOBEC3 activity promotes the survival and evolution of drug-tolerant persister cells during acquired resistance to EGFR inhibitors in lung cancer]]></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/10.1101/2023.07.09.548272v1?rss=1">
<title>
<![CDATA[
Quantifying Cell-State Densities in Single-Cell Phenotypic Landscapes using Mellon 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.09.548272v1?rss=1</link>
<description><![CDATA[
Cell-state density characterizes the distribution of cells along phenotypic landscapes and is crucial for unraveling the mechanisms that drive cellular differentiation, regeneration, and disease. Here, we present Mellon, a novel computational algorithm for high-resolution estimation of cell-state densities from single-cell data. We demonstrate Mellons efficacy by dissecting the density landscape of various differentiating systems, revealing a consistent pattern of high-density regions corresponding to major cell types intertwined with low-density, rare transitory states. Utilizing hematopoietic stem cell fate specification to B-cells as a case study, we present evidence implicating enhancer priming and the activation of master regulators in the emergence of these transitory states. Mellon offers the flexibility to perform temporal interpolation of time-series data, providing a detailed view of cell-state dynamics during the inherently continuous developmental processes. Scalable and adaptable, Mellon facilitates density estimation across various single-cell data modalities, scaling linearly with the number of cells. Our work underscores the importance of cell-state density in understanding the differentiation processes, and the potential of Mellon to provide new insights into the regulatory mechanisms guiding cellular fate decisions.
]]></description>
<dc:creator>Otto, D. J.</dc:creator>
<dc:creator>Jordan, C.</dc:creator>
<dc:creator>Dury, B.</dc:creator>
<dc:creator>Dien, C.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:date>2023-07-10</dc:date>
<dc:identifier>doi:10.1101/2023.07.09.548272</dc:identifier>
<dc:title><![CDATA[Quantifying Cell-State Densities in Single-Cell Phenotypic Landscapes using Mellon]]></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/10.1101/2023.07.10.548412v1?rss=1">
<title>
<![CDATA[
DUX4 is a common driver of immune evasion and immunotherapy failure in metastatic cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.10.548412v1?rss=1</link>
<description><![CDATA[
Cancer immune evasion contributes to checkpoint immunotherapy failure in many patients with metastatic cancers. The embryonic transcription factor DUX4 was recently characterized as a suppressor of interferon-{gamma} signaling and antigen presentation that is aberrantly expressed in a small subset of primary tumors. Here, we report that DUX4 expression is a common feature of metastatic tumors, with [~]10-50% of advanced bladder, breast, kidney, prostate, and skin cancers expressing DUX4. DUX4 expression is significantly associated with immune cell exclusion and decreased objective response to PD-L1 blockade in a large cohort of urothelial carcinoma patients. DUX4 expression is a significant predictor of survival even after accounting for tumor mutational burden and other molecular and clinical features in this cohort, with DUX4 expression associated with a median reduction in survival of over one year. Our data motivate future attempts to develop DUX4 as a biomarker and therapeutic target for checkpoint immunotherapy resistance.
]]></description>
<dc:creator>Pineda, J. M. B.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:date>2023-07-11</dc:date>
<dc:identifier>doi:10.1101/2023.07.10.548412</dc:identifier>
<dc:title><![CDATA[DUX4 is a common driver of immune evasion and immunotherapy failure in metastatic cancers]]></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/10.1101/2023.07.12.548702v1?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/10.1101/2023.07.12.548732v1?rss=1">
<title>
<![CDATA[
Transposable elements regulate thymus development and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.12.548732v1?rss=1</link>
<description><![CDATA[
Transposable elements (TE) are repetitive sequences representing [~]45% of the human and mouse genomes and are highly expressed by medullary thymic epithelial cells (mTEC). In this study, we investigated the role of TEs on T-cell development in the thymus. We performed multi-omic analyses of TEs in human and mouse thymic cells to elucidate their role in T cell development. We report that TE expression in the human thymus is high and shows extensive age- and cell lineage-related variations. TE expression correlates with multiple transcription factors in all cell types of the human thymus. Two cell types express particularly broad TE repertoires: mTECs and plasmacytoid dendritic cells (pDC). In mTECs, transcriptomic data suggest that TEs interact with transcription factors essential for mTEC development and function (e.g., PAX1 and REL), and immunopeptidomic data showed that TEs generate MHC-I-associated peptides implicated in thymocyte education. Notably, AIRE, FEZF2, and CHD4 regulate small yet non-redundant sets of TEs in murine mTECs. Human thymic pDCs homogenously express large numbers of TEs that likely form dsRNA, which can activate innate immune receptors, potentially explaining why thymic pDCs constitutively secrete IFN LJ/{beta}. This study highlights the diversity of interactions between TEs and the adaptive immune system. TEs are genetic parasites, and the two thymic cell types most affected by TEs (mTEcs and pDCs) are essential to establishing central T-cell tolerance. Therefore, we propose that orchestrating TE expression in thymic cells is critical to prevent autoimmunity in vertebrates.
]]></description>
<dc:creator>Larouche, J.-D.</dc:creator>
<dc:creator>Laumont, C. M.</dc:creator>
<dc:creator>Trofimov, A.</dc:creator>
<dc:creator>Vincent, K.</dc:creator>
<dc:creator>Hesnard, L.</dc:creator>
<dc:creator>Brochu, S.</dc:creator>
<dc:creator>Cote, C.</dc:creator>
<dc:creator>Humeau, J.</dc:creator>
<dc:creator>Bonneil, E.</dc:creator>
<dc:creator>Lanoix, J.</dc:creator>
<dc:creator>Durette, C.</dc:creator>
<dc:creator>Gendron, P.</dc:creator>
<dc:creator>Laverdure, J.-P.</dc:creator>
<dc:creator>Richie, E. R.</dc:creator>
<dc:creator>Lemieux, S.</dc:creator>
<dc:creator>Thibault, P.</dc:creator>
<dc:creator>Perreault, C.</dc:creator>
<dc:date>2023-07-14</dc:date>
<dc:identifier>doi:10.1101/2023.07.12.548732</dc:identifier>
<dc:title><![CDATA[Transposable elements regulate thymus development and function]]></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/10.1101/2023.07.17.549345v1?rss=1">
<title>
<![CDATA[
Reanalysis of single-cell RNA sequencing data does not support herpes simplex virus 1 latency in non-neuronal ganglionic cells in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.17.549345v1?rss=1</link>
<description><![CDATA[
Most individuals are latently infected with herpes simplex virus type 1 (HSV-1) and it is well-established that HSV-1 establishes latency in sensory neurons of peripheral ganglia. However, it was recently proposed that latent virus is also present in immune cells recovered from ganglia in a mouse model used for studying latency. Here, we reanalyzed the single-cell RNA sequencing (scRNA-Seq) data that formed the basis for this conclusion. Unexpectedly, off-target priming in 3 scRNA-Seq experiments enabled the detection of non-polyadenylated HSV-1 latency-associated transcript (LAT) intronic RNAs. However, LAT reads were near-exclusively detected in a mixed population of cells undergoing cell death. Specific loss of HSV-1 LAT and neuronal transcripts during quality control filtering indicated widespread destruction of neurons, supporting the presence of contaminating cell-free RNA in other cells following tissue processing. In conclusion, the reported detection of latent HSV-1 in non-neuronal cells is best explained by inaccuracies in the data analyses.
]]></description>
<dc:creator>Ouwendijk, W. J.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Cunningham, A. L.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Koelle, D. M.</dc:creator>
<dc:creator>Kinchington, P. R.</dc:creator>
<dc:creator>Mohr, I.</dc:creator>
<dc:creator>Wilson, A. C.</dc:creator>
<dc:creator>Verjans, G. M. G. M.</dc:creator>
<dc:creator>Depledge, D. P.</dc:creator>
<dc:date>2023-07-18</dc:date>
<dc:identifier>doi:10.1101/2023.07.17.549345</dc:identifier>
<dc:title><![CDATA[Reanalysis of single-cell RNA sequencing data does not support herpes simplex virus 1 latency in non-neuronal ganglionic cells in mice]]></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/10.1101/2023.07.25.550557v1?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/10.1101/2023.07.25.550593v1?rss=1">
<title>
<![CDATA[
Surprising connections between DNA binding and function for the near-complete set of yeast transcription factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.25.550593v1?rss=1</link>
<description><![CDATA[
DNA sequence-specific transcription factors (TFs) modulate transcription and chromatin architecture, acting from regulatory sites in enhancers and promoters of eukaryotic genes. How TFs locate their DNA targets and how multiple TFs cooperate to regulate individual genes is still unclear. Most yeast TFs are thought to regulate transcription via binding to upstream activating sequences, situated within a few hundred base pairs upstream of the regulated gene. While this model has been validated for individual TFs and specific genes, it has not been tested in a systematic way with the large set of yeast TFs. Here, we have integrated information on the binding and expression targets for the near-complete set of yeast TFs. While we found many instances of functional TF binding sites in upstream regulatory regions, we found many more instances that do not fit this model. In many cases, rapid TF depletion affects gene expression where there is no detectable binding of that TF to the upstream region of the affected gene. In addition, for most TFs, only a small fraction of bound TFs regulates the nearby gene, showing that TF binding does not automatically correspond to regulation of the linked gene. Finally, we found that only a small percentage of TFs are exclusively strong activators or repressors with most TFs having dual function. Overall, our comprehensive mapping of TF binding and regulatory targets have both confirmed known TF relationships and revealed surprising properties of TF function.
]]></description>
<dc:creator>Mahendrawada, L.</dc:creator>
<dc:creator>Warfield, L.</dc:creator>
<dc:creator>Donczew, R.</dc:creator>
<dc:creator>hahn, s.</dc:creator>
<dc:date>2023-07-27</dc:date>
<dc:identifier>doi:10.1101/2023.07.25.550593</dc:identifier>
<dc:title><![CDATA[Surprising connections between DNA binding and function for the near-complete set of yeast transcription factors]]></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/10.1101/2023.07.28.551016v1?rss=1">
<title>
<![CDATA[
A CRISPR screen of HIV dependency factors reveals CCNT1 is non-essential in T cells but required for HIV-1 reactivation from latency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.28.551016v1?rss=1</link>
<description><![CDATA[
We sought to explore the hypothesis that host factors required for HIV-1 replication also play a role in latency reversal. Using a CRISPR gene library of putative HIV dependency factors, we performed a screen to identify genes required for latency reactivation. We identified several HIV-1 dependency factors that play a key role in HIV-1 latency reactivation including ELL, UBE2M, TBL1XR1, HDAC3, AMBRA1, and ALYREF. Knockout of Cyclin T1 (CCNT1), a component of the P-TEFb complex important for transcription elongation, was the top hit in the screen and had the largest effect on HIV latency reversal with a wide variety of latency reversal agents. Moreover, CCNT1 knockout prevents latency reactivation in a primary CD4+ T cell model of HIV latency without affecting activation of these cells. RNA sequencing data showed that CCNT1 regulates HIV-1 proviral genes to a larger extent than any other host gene and had no significant effects on RNA transcripts in primary T cells after activation. We conclude that CCNT1 function is redundant in T cells but is absolutely required for HIV latency reversal.
]]></description>
<dc:creator>Hafer, T. L.</dc:creator>
<dc:creator>Felton, A.</dc:creator>
<dc:creator>Delgado, Y.</dc:creator>
<dc:creator>Srinivasan, H.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2023-07-28</dc:date>
<dc:identifier>doi:10.1101/2023.07.28.551016</dc:identifier>
<dc:title><![CDATA[A CRISPR screen of HIV dependency factors reveals CCNT1 is non-essential in T cells but required for HIV-1 reactivation from latency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.27.550871v1?rss=1">
<title>
<![CDATA[
TGF-β broadly modifies rather than specifically suppresses reactivated memory CD8 T cells in a dose-dependent manner 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.27.550871v1?rss=1</link>
<description><![CDATA[
Transforming growth factor {beta} (TGF-{beta}) directly acts on naive, effector and memory T cells to control cell fate decisions, which was shown using genetic abrogation of TGF-{beta} signaling. TGF-{beta} availability is altered by infections and cancer, however the dose-dependent effects of TGF-{beta} on memory CD8 T cell (Tmem) reactivation are still poorly defined. We examined how activation and TGF-{beta} signals interact to shape the functional outcome of Tmem reactivation. We found that TGF-{beta} could suppress cytotoxicity in a manner that was inversely proportional to the strength of the activating TCR or pro-inflammatory signals. In contrast, even high doses of TGF-{beta} had a comparatively modest effect on IFN-{gamma} expression in the context of weak and strong reactivation signals. Since CD8 Tmem may not always receive TGF-{beta} signals concurrently with reactivation, we also explored whether the temporal order of reactivation versus TGF-{beta} signals is of importance. We found that exposure to TGF-{beta} prior to as well as after an activation event were both sufficient to reduce cytotoxic effector function. Concurrent ATAC-seq and RNA-seq analysis revealed that TGF-{beta} altered [~]10% of the regulatory elements induced by reactivation and also elicited transcriptional changes indicative of broadly modulated functional properties. We confirmed some changes on the protein level and found that TGF-{beta}-induced expression of CCR8 was inversely proportional to the strength of the reactivating TCR signal. Together, our data suggest that TGF-{beta} is not simply suppressing CD8 Tmem, but modifies functional and chemotactic properties in context of their reactivation signals and in a dose-dependent manner.
]]></description>
<dc:creator>Taber, A.</dc:creator>
<dc:creator>Konecny, A.</dc:creator>
<dc:creator>Scott-Browne, J.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:date>2023-07-29</dc:date>
<dc:identifier>doi:10.1101/2023.07.27.550871</dc:identifier>
<dc:title><![CDATA[TGF-β broadly modifies rather than specifically suppresses reactivated memory CD8 T cells in a dose-dependent manner]]></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/10.1101/2023.07.27.550902v1?rss=1">
<title>
<![CDATA[
A multiplex, prime editing framework for identifying drug resistance variants at scale 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.27.550902v1?rss=1</link>
<description><![CDATA[
CRISPR-based genome editing has revolutionized functional genomics, enabling screens in which thousands of perturbations of either gene expression or primary genome sequence can be competitively assayed in single experiments. However, for libraries of specific mutations, a challenge of CRISPR-based screening methods such as saturation genome editing is that only one region (e.g. one exon) can be studied per experiment. Here we describe prime-SGE ("prime saturation genome editing"), a new framework based on prime editing, in which libraries of specific mutations can be installed into genes throughout the genome and functionally assessed in a single, multiplex experiment. Prime-SGE is based on quantifying the abundance of prime editing guide RNAs (pegRNAs) in the context of a functional selection, rather than quantifying the mutations themselves. We apply prime-SGE to assay thousands of single nucleotide changes in eight oncogenes for their ability to confer drug resistance to three EGFR tyrosine kinase inhibitors. Although currently restricted to positive selection screens by the limited efficiency of prime editing, our strategy opens the door to the possibility of functionally assaying vast numbers of precise mutations at locations throughout the genome.
]]></description>
<dc:creator>Chardon, F. M.</dc:creator>
<dc:creator>Suiter, C.</dc:creator>
<dc:creator>Daza, R.</dc:creator>
<dc:creator>Smith, N. T.</dc:creator>
<dc:creator>Parrish, P.</dc:creator>
<dc:creator>McDiarmid, T.</dc:creator>
<dc:creator>Lalanne, J.-B.</dc:creator>
<dc:creator>Martin, B. K.</dc:creator>
<dc:creator>Calderon, D.</dc:creator>
<dc:creator>Ellison, A.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Starita, L. M.</dc:creator>
<dc:date>2023-07-30</dc:date>
<dc:identifier>doi:10.1101/2023.07.27.550902</dc:identifier>
<dc:title><![CDATA[A multiplex, prime editing framework for identifying drug resistance variants at scale]]></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/10.1101/2023.07.31.551363v1?rss=1">
<title>
<![CDATA[
A robust method for measuring aminoacylation through tRNA-Seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.31.551363v1?rss=1</link>
<description><![CDATA[
Current methods to quantify the fraction of aminoacylated tRNAs, also known as the tRNA charge, are limited by issues with either low throughput, precision, and/or accuracy. Here, we present an optimized charge tRNA-Seq method that combines previous developments with newly described approaches to establish a protocol for precise and accurate tRNA charge measurements. We verify that this protocol provides robust quantification of tRNA aminoacylation and we provide an end-to-end method that scales to hundreds of samples including software for data processing. Additionally, we show that this method supports measurements of relative tRNA expression levels and can be used to infer tRNA modifications through reverse transcription misincorporations, thereby supporting multipurpose applications in tRNA biology.
]]></description>
<dc:creator>Davidsen, K.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:date>2023-08-01</dc:date>
<dc:identifier>doi:10.1101/2023.07.31.551363</dc:identifier>
<dc:title><![CDATA[A robust method for measuring aminoacylation through tRNA-Seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.31.551037v1?rss=1">
<title>
<![CDATA[
Jointly modeling deep mutational scans identifies shifted mutational effects among SARS-CoV-2 spike homologs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.31.551037v1?rss=1</link>
<description><![CDATA[
Deep mutational scanning (DMS) is a high-throughput experimental technique that measures the effects of thousands of mutations to a protein. These experiments can be performed on multiple homologs of a protein or on the same protein selected under multiple conditions. It is often of biological interest to identify mutations with shifted effects across homologs or conditions. However, it is challenging to determine if observed shifts arise from biological signal or experimental noise. Here, we describe a method for jointly inferring mutational effects across multiple DMS experiments while also identifying mutations that have shifted in their effects among experiments. A key aspect of our method is to regularize the inferred shifts, so that they are nonzero only when strongly supported by the data. We apply this method to DMS experiments that measure how mutations to spike proteins from SARS-CoV-2 variants (Delta, Omicron BA.1, and Omicron BA.2) affect cell entry. Most mutational effects are conserved between these spike homologs, but a fraction have markedly shifted. We experimentally validate a subset of the mutations inferred to have shifted effects, and confirm differences of >1,000-fold in the impact of the same mutation on spike-mediated viral infection across spikes from different SARS-CoV-2 variants. Overall, our work establishes a general approach for comparing sets of DMS experiments to identify biologically important shifts in mutational effects.

Significance StatementAmino-acid mutations to a protein have effects that can shift as the protein evolves or is put under new selective pressure. The effects of amino-acid mutations to a specific protein under a defined selective pressure can be measured by deep mutational scanning experiments. Here, we devise an approach to quantify shifts in mutational effects between experiments performed on different homologs (i.e. variants) of the same protein, or on the same protein selected under different conditions. We use this approach to compare experiments performed on three homologs of SARS-CoV-2 spike, identifying mutations that have shifted in their effect on spike-mediated viral infection by >1,000 fold across SARS-CoV-2 variants.
]]></description>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>DeWitt, W. S.</dc:creator>
<dc:date>2023-08-02</dc:date>
<dc:identifier>doi:10.1101/2023.07.31.551037</dc:identifier>
<dc:title><![CDATA[Jointly modeling deep mutational scans identifies shifted mutational effects among SARS-CoV-2 spike homologs]]></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/10.1101/2023.08.02.551697v1?rss=1">
<title>
<![CDATA[
Unraveling the Global Proteome and Phosphoproteome of Prostate Cancer Patient-Derived Xenografts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.02.551697v1?rss=1</link>
<description><![CDATA[
Resistance to androgen deprivation therapies leads to metastatic castration-resistant prostate cancer (mCRPC) of adenocarcinoma (AdCa) origin that can transform to emergent aggressive variant prostate cancer (AVPC) which has neuroendocrine (NE)-like features. To this end, we used LuCaP patient-derived xenograft (PDX) tumors, clinically relevant models that reflects and retains key features of the tumor from advanced prostate cancer patients. Here we performed proteome and phosphoproteome characterization of 48 LuCaP PDX tumors and identified over 94,000 peptides and 9,700 phosphopeptides corresponding to 7,738 proteins. When we compared 15 NE versus 33 AdCa PDX samples, we identified 309 unique proteins and 476 unique phosphopeptides that were significantly altered and corresponded to proteins that are known to distinguish these two phenotypes. Assessment of protein and RNA concordance from these tumors revealed increased dissonance in transcriptionally regulated proteins in NE and metabolite interconversion enzymes in AdCa.
]]></description>
<dc:creator>Sychev, Z. E.</dc:creator>
<dc:creator>Day, A.</dc:creator>
<dc:creator>Bergom, H. E.</dc:creator>
<dc:creator>Larson, G.</dc:creator>
<dc:creator>Ali, A.</dc:creator>
<dc:creator>Ludwig, M.</dc:creator>
<dc:creator>Boytim, E.</dc:creator>
<dc:creator>Ilsa, C.</dc:creator>
<dc:creator>Corey, E.</dc:creator>
<dc:creator>Plymate, S. R.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Hwang, J. H.</dc:creator>
<dc:creator>Drake, J. M.</dc:creator>
<dc:date>2023-08-05</dc:date>
<dc:identifier>doi:10.1101/2023.08.02.551697</dc:identifier>
<dc:title><![CDATA[Unraveling the Global Proteome and Phosphoproteome of Prostate Cancer Patient-Derived Xenografts]]></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/10.1101/2023.08.17.553668v1?rss=1">
<title>
<![CDATA[
Targeting CD19-positive lymphomas with the antibody-drug conjugate (ADC) loncastuximab tesirine: preclinical evidence as single agent and as combinatorial approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.17.553668v1?rss=1</link>
<description><![CDATA[
PurposeAntibody-drug conjugates (ADCs) represent one of the most successful therapeutic approaches introduced in clinical practice in the last years. Loncastuximab tesirine (ADCT-402) is a CD19 targeting ADC, in which the antibody is conjugated through a protease cleavable dipeptide linker to a pyrrolobenzodiazepine (PBD) dimer warhead (SG3199). Based on the results of a phase 2 study, loncastuximab tesirine was recently approved for adult patients with relapsed/refractory large B-cell lymphoma.

Experimental DesignWe assessed the activity of loncastuximab tesirine in in vitro and in vivo models of lymphomas, correlated its activity with CD19 expression levels and identified combination partners providing synergy with loncastuximab tesirine.

ResultsLoncastuximab tesirine was tested across 60 lymphoma cell lines. Loncastuximab tesirine has strong cytotoxic activity in B-cell lymphoma cell lines and the in vitro activity is correlated with CD19 expression level and with intrinsic sensitivity of cell lines to the ADCs warhead.

Loncastuximab tesirine was more potent than other anti-CD19 ADCs (coltuximab ravtansine, huB4-DGN462), albeit the pattern of activity across cell lines was correlated. Loncastuximab tesirine activity also largely correlated with cell line sensitivity to R-CHOP.

Combinatorial in vitro and in vivo experiments identified the benefit of adding loncastuximab tesirine to other agents, especially BCL2 and PI3K inhibitors.

ConclusionsOur data support the further development of loncastuximab tesirine as single agent and in combination for patients affected by mature B-cell neoplasms. The results also highlight the importance of CD19 expression, and the existence of lymphoma populations characterized by resistance to multiple therapies.
]]></description>
<dc:creator>Tarantelli, C.</dc:creator>
<dc:creator>Wald, D.</dc:creator>
<dc:creator>Munz, N.</dc:creator>
<dc:creator>Spriano, F.</dc:creator>
<dc:creator>Bruscaggin, A.</dc:creator>
<dc:creator>Cannas, E.</dc:creator>
<dc:creator>Cascione, L.</dc:creator>
<dc:creator>Gaudio, E.</dc:creator>
<dc:creator>Arribas, A. J.</dc:creator>
<dc:creator>Manjappa, S.</dc:creator>
<dc:creator>Golino, G.</dc:creator>
<dc:creator>Scalise, L.</dc:creator>
<dc:creator>Zucca, E.</dc:creator>
<dc:creator>Stathis, A.</dc:creator>
<dc:creator>Van Berkel, P. H.</dc:creator>
<dc:creator>Rossi, D.</dc:creator>
<dc:creator>Caimi, P. F.</dc:creator>
<dc:creator>Zammarchi, F.</dc:creator>
<dc:creator>Bertoni, F.</dc:creator>
<dc:date>2023-08-18</dc:date>
<dc:identifier>doi:10.1101/2023.08.17.553668</dc:identifier>
<dc:title><![CDATA[Targeting CD19-positive lymphomas with the antibody-drug conjugate (ADC) loncastuximab tesirine: preclinical evidence as single agent and as combinatorial approach]]></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/10.1101/2023.08.20.554063v1?rss=1">
<title>
<![CDATA[
KinCytE- a Kinase to Cytokine Explorer to Identify Molecular Regulators and Potential Therapeutic Opportunities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.20.554063v1?rss=1</link>
<description><![CDATA[
Cytokines and chemokines are secreted proteins that regulate various biological processes, such as inflammation, immune response, and cell differentiation. Therefore, disruption of signaling pathways involving these proteins has been linked to a range of diseases, including cancer. However, targeting individual cytokines, chemokines, or their receptors is challenging due to their regulatory redundancy and incomplete understanding of their signaling networks. To transform these difficult-to-drug targets into a pharmacologically manageable class, we developed a web-based platform called KinCytE. This platform was designed to link the effects of kinase inhibitors, a well-established class of drugs, with cytokine and chemokine release and signaling networks. The resulting KinCytE platform enables users to investigate protein kinases that regulate specific cytokines or chemokines, generate a ranked list of FDA-approved kinase inhibitors that affect cytokine/chemokine activity, and explore and visualize cytokine signaling network thus facilitating drugging this challenging target class. KinCytE is freely accessible via https://atlas.fredhutch.org/kincyte.
]]></description>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Osborne, S.</dc:creator>
<dc:creator>Zager, M.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.20.554063</dc:identifier>
<dc:title><![CDATA[KinCytE- a Kinase to Cytokine Explorer to Identify Molecular Regulators and Potential Therapeutic Opportunities]]></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/10.1101/2023.08.27.555013v1?rss=1">
<title>
<![CDATA[
MADDD-seq, a novel massively parallel sequencing tool for simultaneous detection of DNA damage and mutations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.27.555013v1?rss=1</link>
<description><![CDATA[
Our genome is exposed to a wide variety of DNA-damaging agents. If left unrepaired, this damage can be fixed into mutations that promote carcinogenesis and the development of genetically inherited diseases. As a result, it is crucial that we can detect DNA damage and mutations with exquisite sensitivity. Here, we describe a modified version of double barcoding sequencing technology termed Mutation And DNA Damage Detection-seq (MADDD-seq) that can detect DNA damage and mutations simultaneously, with a single assay. To demonstrate the utility of MADDD-seq as a multifunctional detection tool, we treated yeast cells with a DNA-damaging agent and tracked the presence of DNA damage and mutations over a 24-hour timespan. These experiments allowed us to identify thousands of adducts and mutations in a single sequencing run and expose the kinetics of DNA repair and mutagenesis in remarkable detail.
]]></description>
<dc:creator>Vermulst, M.</dc:creator>
<dc:creator>Paskvan, S.</dc:creator>
<dc:creator>Chung, C.</dc:creator>
<dc:creator>Franke, K.</dc:creator>
<dc:creator>Clegg, N.</dc:creator>
<dc:creator>Madeoy, J.</dc:creator>
<dc:creator>Long, A. N.</dc:creator>
<dc:creator>Gout, J.-F.</dc:creator>
<dc:creator>Bielas, J. H.</dc:creator>
<dc:date>2023-08-27</dc:date>
<dc:identifier>doi:10.1101/2023.08.27.555013</dc:identifier>
<dc:title><![CDATA[MADDD-seq, a novel massively parallel sequencing tool for simultaneous detection of DNA damage and mutations]]></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/10.1101/2023.08.30.555623v1?rss=1">
<title>
<![CDATA[
Unraveling human hematopoietic progenitor cell diversity through association with intrinsic regulatory factors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.30.555623v1?rss=1</link>
<description><![CDATA[
Hematopoietic stem and progenitor cell (HSPC) transplantation is an essential therapy for hematological conditions, but finer definitions of human HSPC subsets with associated function could enable better tuning of grafts and more routine, lower-risk application. To deeply phenotype HSPCs, following a screen of 328 antigens, we quantified 41 surface proteins and functional regulators on millions of CD34+ and CD34- cells, spanning four primary human hematopoietic tissues: bone marrow, mobilized peripheral blood, cord blood, and fetal liver. We propose more granular definitions of HSPC subsets and provide new, detailed differentiation trajectories of erythroid and myeloid lineages. These aspects of our revised human hematopoietic model were validated with corresponding epigenetic analysis and in vitro clonal differentiation assays. Overall, we demonstrate the utility of using molecular regulators as surrogates for cellular identity and functional potential, providing a framework for description, prospective isolation, and cross-tissue comparison of HSPCs in humans.
]]></description>
<dc:creator>Favaro, P.</dc:creator>
<dc:creator>Glass, D. R.</dc:creator>
<dc:creator>Borges, L.</dc:creator>
<dc:creator>Reemar, R. B.</dc:creator>
<dc:creator>Reynolds, W.</dc:creator>
<dc:creator>Ho, D.</dc:creator>
<dc:creator>Bruce, T.</dc:creator>
<dc:creator>Tebaykin, D.</dc:creator>
<dc:creator>Scalon, V. M.</dc:creator>
<dc:creator>Shestopalov, I.</dc:creator>
<dc:creator>Bendall, S. C.</dc:creator>
<dc:date>2023-08-31</dc:date>
<dc:identifier>doi:10.1101/2023.08.30.555623</dc:identifier>
<dc:title><![CDATA[Unraveling human hematopoietic progenitor cell diversity through association with intrinsic regulatory factors]]></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/10.1101/2023.08.30.555598v1?rss=1">
<title>
<![CDATA[
Chromatin Profiling of CBFA2T3-GLIS2 AMLs Identifies Key Transcription Factor Dependencies and BRG1 Inhibition as a Novel Therapeutic Strategy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.30.555598v1?rss=1</link>
<description><![CDATA[
Oncogenic fusions involving transcription factors are present in the majority of pediatric leukemias; however, the context-specific mechanisms they employ to drive cancer remain poorly understood. CBFA2T3-GLIS2 (C/G) fusions occur in treatment-refractory acute myeloid leukemias and are restricted to young children. To understand how the C/G fusion drives oncogenesis we applied CUT&RUN chromatin profiling to an umbilical cord blood/endothelial cell (EC) co-culture model of C/G AML that recapitulates the biology of this malignancy. We find C/G fusion binding is mediated by its zinc finger domains. Integration of fusion binding sites in C/G- transduced cells with Polycomb Repressive Complex 2 (PRC2) sites in control cord blood cells identifies MYCN, ZFPM1, ZBTB16 and LMO2 as direct C/G targets. Transcriptomic analysis of a large pediatric AML cohort shows that these genes are upregulated in C/G patient samples. Single cell RNA-sequencing of umbilical cord blood identifies a population of megakaryocyte precursors that already express many of these genes despite lacking the fusion. By integrating CUT&RUN data with CRISPR dependency screens we identify BRG1/SMARCA4 as a vulnerability in C/G AML. BRG1 profiling in C/G patient-derived cell lines shows that the CBFA2T3 locus is a binding site, and treatment with clinically-available BRG1 inhibitors reduces fusion levels and downstream C/G targets including N-MYC, resulting in C/G leukemia cell death and extending survival in a murine xenograft model.
]]></description>
<dc:creator>Kaonis, S.</dc:creator>
<dc:creator>Smith, J. L.</dc:creator>
<dc:creator>Katiyar, N.</dc:creator>
<dc:creator>Merrill, M.</dc:creator>
<dc:creator>Hylkema, T.</dc:creator>
<dc:creator>Namciu, S.</dc:creator>
<dc:creator>Le, Q.</dc:creator>
<dc:creator>Babaeva, E.</dc:creator>
<dc:creator>Ikeda, T.</dc:creator>
<dc:creator>Morris, S. M.</dc:creator>
<dc:creator>Girard, E.</dc:creator>
<dc:creator>Furuyama, S.</dc:creator>
<dc:creator>Ries, R.</dc:creator>
<dc:creator>Bernstein, I.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Meers, M. P.</dc:creator>
<dc:creator>Hadland, B. K.</dc:creator>
<dc:creator>Sarthy, J. F.</dc:creator>
<dc:date>2023-09-01</dc:date>
<dc:identifier>doi:10.1101/2023.08.30.555598</dc:identifier>
<dc:title><![CDATA[Chromatin Profiling of CBFA2T3-GLIS2 AMLs Identifies Key Transcription Factor Dependencies and BRG1 Inhibition as a Novel Therapeutic Strategy]]></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/10.1101/2023.09.01.554391v1?rss=1">
<title>
<![CDATA[
Versatile Tissue-Injectable Hydrogels with Extended Hydrolytic Release of Bioactive Protein Therapeutics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.01.554391v1?rss=1</link>
<description><![CDATA[
Hydrogels generally have broad utilization in healthcare due to their tunable structures, high water content, and inherent biocompatibility. FDA-approved applications of hydrogels include spinal cord regeneration, skin fillers, and local therapeutic delivery. Drawbacks exist in the clinical hydrogel space, largely pertaining to inconsistent therapeutic exposure, short-lived release windows, and difficulties inserting the polymer into tissue. In this study, we engineered injectable, biocompatible hydrogels that function as a local protein therapeutic depot with a high degree of user-customizability. We showcase a PEG-based hydrogel functionalized with bioorthogonal strain-promoted azide-alkyne cycloaddition (SPAAC) handles for its polymerization and functionalization with a variety of payloads. Small-molecule and protein cargos, including chemokines and antibodies, were site-specifically modified with hydrolysable "azidoesters" of varying hydrophobicity via direct chemical conjugation or sortase-mediated transpeptidation. These hydrolysable esters afforded extended release of payloads linked to our hydrogels beyond diffusion; with timescales spanning days to months dependent on ester hydrophobicity. Injected hydrogels polymerize in situ and remain in tissue over extended periods of time. Hydrogel-delivered protein payloads elicit biological activity after being modified with SPAAC-compatible linkers, as demonstrated by the successful recruitment of murine T-cells to a mouse melanoma model by hydrolytically released murine CXCL10. These results highlight a highly versatile, customizable hydrogel-based delivery system for local delivery of protein therapeutics with payload release profiles appropriate for a variety of clinical needs.

Translational ImpactWe developed injectable hydrogels that provide loco-regional, controlled release of protein therapeutics. Local delivery is especially suitable for potent, protein-based drugs like chemokines and monoclonal antibodies whose systemic toxicities can be life threatening. Our hydrogel is equipped with slowly hydrolyzing linkers for transient payload coupling, prolonging its therapeutic window and minimizing the need for repeat surgeries in a clinical setting. A range of release profiles, spanning days to over a month, and a broad compatibility to therapeutically relevant, recombinant proteins provide clinicians with flexible therapeutic options to suit a variety of circumstances.
]]></description>
<dc:creator>Nealy, E. S.</dc:creator>
<dc:creator>Reed, S. J.</dc:creator>
<dc:creator>Badeau, B. A.</dc:creator>
<dc:creator>Adelmund, S. M.</dc:creator>
<dc:creator>Shadish, J. A.</dc:creator>
<dc:creator>Pakiam, F. J.</dc:creator>
<dc:creator>Girard, E. J.</dc:creator>
<dc:creator>Price, J. P.</dc:creator>
<dc:creator>Mhyre, A. J.</dc:creator>
<dc:creator>Kalia, V.</dc:creator>
<dc:creator>Sarkar, S.</dc:creator>
<dc:creator>DeForest, C. A.</dc:creator>
<dc:creator>Olson, J. M.</dc:creator>
<dc:date>2023-09-02</dc:date>
<dc:identifier>doi:10.1101/2023.09.01.554391</dc:identifier>
<dc:title><![CDATA[Versatile Tissue-Injectable Hydrogels with Extended Hydrolytic Release of Bioactive Protein Therapeutics]]></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/10.1101/2023.09.05.556399v1?rss=1">
<title>
<![CDATA[
Host cell glycosylation selects for infection with CCR5- versus CXCR4-tropic HIV-1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.05.556399v1?rss=1</link>
<description><![CDATA[
HIV-1 infection involves a selection bottleneck that leads to transmission of one or a few HIV variants, which nearly always use CCR5 as the coreceptor (R5 viruses) for viral entry as opposed to CXCR4 (X4 viruses). The host properties that drive this selection are not well understood and may hold keys to factors that govern HIV susceptibility. In this report, we identified SLC35A2, a transporter of UDP-galactose, as a candidate X4-specific restriction factor in CRISPR-knockout screens in primary target CD4+ T cells. SLC35A2 inactivation in CD4+ T cells, which resulted in truncation of glycans due to the absence of galactose, not only increased X4 infection levels, but also consistently decreased infection levels of R5 HIV strains. Single cycle infections demonstrated that the effect is host cell dependent. SLC35A2 is expressed in CD4+ T cells at different tissue sites, with high levels in the genital tract - the site of most HIV infections. These data support a role for a host cell protein that regulates glycan structure on HIV infection, with enhanced R5 infection but reduced X4 infection associated with SLC35A2-mediated glycosylation. Host cell glycosylation may therefore contribute to R5 selection and host susceptibility during HIV transmission.
]]></description>
<dc:creator>Itell, H. L.</dc:creator>
<dc:creator>Humes, D.</dc:creator>
<dc:creator>Baumgarten, N. E.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2023-09-05</dc:date>
<dc:identifier>doi:10.1101/2023.09.05.556399</dc:identifier>
<dc:title><![CDATA[Host cell glycosylation selects for infection with CCR5- versus CXCR4-tropic HIV-1]]></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/10.1101/2023.09.10.555215v1?rss=1">
<title>
<![CDATA[
Whole Genome Sequencing Based Analysis of Inflammation Biomarkers in the Trans-Omics for Precision Medicine (TOPMed) Consortium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.10.555215v1?rss=1</link>
<description><![CDATA[
Inflammation biomarkers can provide valuable insight into the role of inflammatory processes in many diseases and conditions. Sequencing based analyses of such biomarkers can also serve as an exemplar of the genetic architecture of quantitative traits. To evaluate the biological insight, which can be provided by a multi-ancestry, whole-genome based association study, we performed a comprehensive analysis of 21 inflammation biomarkers from up to 38,465 individuals with whole-genome sequencing from the Trans-Omics for Precision Medicine (TOPMed) program. We identified 22 distinct single-variant associations across 6 traits - E-selectin, intercellular adhesion molecule 1, interleukin-6, lipoprotein-associated phospholipase A2 activity and mass, and P-selectin - that remained significant after conditioning on previously identified associations for these inflammatory biomarkers. We further expanded upon known biomarker associations by pairing the single-variant analysis with a rare variant set-based analysis that further identified 19 significant rare variant set-based associations with 5 traits. These signals were distinct from both significant single variant association signals within TOPMed and genetic signals observed in prior studies, demonstrating the complementary value of performing both single and rare variant analyses when analyzing quantitative traits. We also confirm several previously reported signals from semi-quantitative proteomics platforms. Many of these signals demonstrate the extensive allelic heterogeneity and ancestry-differentiated variant-trait associations common for inflammation biomarkers, a characteristic we hypothesize will be increasingly observed with well-powered, large-scale analyses of complex traits.
]]></description>
<dc:creator>Jiang, M.-Z.</dc:creator>
<dc:creator>Gaynor, S. M.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Van Buren, E.</dc:creator>
<dc:creator>Stilp, A.</dc:creator>
<dc:creator>Buth, E.</dc:creator>
<dc:creator>Wang, F. F.</dc:creator>
<dc:creator>Manansala, R.</dc:creator>
<dc:creator>Gogarten, S. M.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Polfus, L. M.</dc:creator>
<dc:creator>Salimi, S.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Pankratz, N.</dc:creator>
<dc:creator>Yanek, L. R.</dc:creator>
<dc:creator>Durda, P.</dc:creator>
<dc:creator>Tracy, R. P.</dc:creator>
<dc:creator>Rich, S. S.</dc:creator>
<dc:creator>Rotter, J. I.</dc:creator>
<dc:creator>Mitchell, B. D.</dc:creator>
<dc:creator>Lewis, J. P.</dc:creator>
<dc:creator>Psaty, B. M.</dc:creator>
<dc:creator>Pratte, K. A.</dc:creator>
<dc:creator>Silverman, E. K.</dc:creator>
<dc:creator>Kaplan, R. C.</dc:creator>
<dc:creator>Avery, C.</dc:creator>
<dc:creator>North, K.</dc:creator>
<dc:creator>Mathias, R. A.</dc:creator>
<dc:creator>Faraday, N.</dc:creator>
<dc:creator>Lin, H.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Carson, A. P.</dc:creator>
<dc:creator>Norwood, A. F.</dc:creator>
<dc:creator>Gibbs, R. A.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Lundin, J.</dc:creator>
<dc:creator>Peters, U.</dc:creator>
<dc:creator>Dupuis, J.</dc:creator>
<dc:creator>Hou, L.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Benjamin, E. J.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Bowler, R. P.</dc:creator>
<dc:creator>Lin, X.</dc:creator>
<dc:creator>Auer, P. L.</dc:creator>
<dc:creator>Raf</dc:creator>
<dc:date>2023-09-12</dc:date>
<dc:identifier>doi:10.1101/2023.09.10.555215</dc:identifier>
<dc:title><![CDATA[Whole Genome Sequencing Based Analysis of Inflammation Biomarkers in the Trans-Omics for Precision Medicine (TOPMed) Consortium]]></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/10.1101/2023.09.11.557289v1?rss=1">
<title>
<![CDATA[
Position-independent refinement of vagus motor neuron wiring 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.11.557289v1?rss=1</link>
<description><![CDATA[
Motor neurons in the central nervous system often lie in a continuous topographic map, where neurons that innervate different body parts are spatially intermingled. This is the case for the efferent neurons of the vagus nerve, which innervate diverse muscle and organ targets in the head and viscera for brain-body communication. It remains elusive how neighboring motor neurons with different fixed peripheral axon targets develop the separate somatodendritic (input) connectivity they need to generate spatially precise body control. Here we show that vagus motor neurons in the zebrafish indeed generate spatially appropriate peripheral responses to focal sensory stimulation even when they are transplanted into ectopic positions within the topographic map, indicating that circuit refinement occurs after the establishment of coarse topography. Refinement depends on motor neuron synaptic transmission, suggesting that an experience-dependent periphery-to-brain feedback mechanism establishes specific input connectivity amongst intermingled motor populations.
]]></description>
<dc:creator>Kaneko, T.</dc:creator>
<dc:creator>Boulanger-Weill, J.</dc:creator>
<dc:creator>Isabella, A. J.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:date>2023-09-13</dc:date>
<dc:identifier>doi:10.1101/2023.09.11.557289</dc:identifier>
<dc:title><![CDATA[Position-independent refinement of vagus motor neuron wiring]]></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/10.1101/2023.09.13.557576v1?rss=1">
<title>
<![CDATA[
Structure and dynamics of the contractile vacuole complex in Tetrahymena thermophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.13.557576v1?rss=1</link>
<description><![CDATA[
The contractile vacuole complex (CVC) is a dynamic and morphologically complex membrane organelle, comprised of a large vesicle (bladder) linked with a tubular reticulum (spongiome). CVCs provide key osmoregulatory roles across diverse eukaryotic lineages, but probing the mechanisms underlying the structure and function is hampered by the limited tools available for in vivo analysis. In the experimentally tractable ciliate Tetrahymena thermophila, we describe four proteins that, as endogenously tagged constructs, localize specifically to distinct CVC zones. The DOPEY homolog Dop1p and the CORVET subunit Vps8Dp localize both to the bladder and spongiome but with different local distributions that are sensitive to osmotic perturbation, while the lipid scramblase Scr7p co-localizes with Vps8Dp. The H+- ATPase subunit Vma4 is spongiome-specific. The live imaging permitted by these probes revealed dynamics at multiple scales including rapid exchange of CVC-localized and soluble protein pools vs. lateral diffusion in the spongiome, spongiome extension and branching, and CVC formation during mitosis. While the association with DOP1 and VPS8D implicate the CVC in endosomal trafficking, both the bladder and spongiome are isolated from bulk endocytic input.

Summary statementIn the ciliate Tetrahymena thermophila, four proteins are shown to provide markers for different zones of the contractile vacuole complex. They shed light on its formation and maintenance by enabling in vivo analysis of its dynamics.
]]></description>
<dc:creator>Cheng, C.-Y.</dc:creator>
<dc:creator>Romero, D. P.</dc:creator>
<dc:creator>Zoltner, M.</dc:creator>
<dc:creator>Yao, M.-C.</dc:creator>
<dc:creator>Turkewitz, A.</dc:creator>
<dc:date>2023-09-14</dc:date>
<dc:identifier>doi:10.1101/2023.09.13.557576</dc:identifier>
<dc:title><![CDATA[Structure and dynamics of the contractile vacuole complex in Tetrahymena thermophila]]></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/10.1101/2023.09.11.557241v1?rss=1">
<title>
<![CDATA[
Regulation of interferon signaling by transposon exonization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.11.557241v1?rss=1</link>
<description><![CDATA[
SummaryInnate immune signaling is essential for clearing pathogens and damaged cells, and must be tightly regulated to avoid excessive inflammation or autoimmunity. Here, we found that the alternative splicing of exons derived from transposable elements is a key mechanism controlling immune signaling in human cells. By analyzing long-read transcriptome datasets, we identified numerous transposon exonization events predicted to generate functional protein variants of immune genes, including the type I interferon receptor IFNAR2. We demonstrated that the transposon-derived isoform of IFNAR2 is more highly expressed than the canonical isoform in almost all tissues, and functions as a decoy receptor that potently inhibits interferon signaling including in cells infected with SARS-CoV-2. Our findings uncover a primate-specific axis controlling interferon signaling and show how a transposon exonization event can be co-opted for immune regulation.
]]></description>
<dc:creator>Pasquesi, G. I. M.</dc:creator>
<dc:creator>Allen, H.</dc:creator>
<dc:creator>Ivancevic, A.</dc:creator>
<dc:creator>Barbachano-Guerrero, A.</dc:creator>
<dc:creator>Joyner, O.</dc:creator>
<dc:creator>Guo, K.</dc:creator>
<dc:creator>Simpson, D. M.</dc:creator>
<dc:creator>Gapin, K.</dc:creator>
<dc:creator>Horton, I.</dc:creator>
<dc:creator>Nguyen, L.</dc:creator>
<dc:creator>Yang, Q.</dc:creator>
<dc:creator>Warren, C. J.</dc:creator>
<dc:creator>Florea, L. D.</dc:creator>
<dc:creator>Bitler, B. G.</dc:creator>
<dc:creator>Santiago, M. L.</dc:creator>
<dc:creator>Sawyer, S. L.</dc:creator>
<dc:creator>Chuong, E. B.</dc:creator>
<dc:date>2023-09-15</dc:date>
<dc:identifier>doi:10.1101/2023.09.11.557241</dc:identifier>
<dc:title><![CDATA[Regulation of interferon signaling by transposon exonization]]></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/10.1101/2023.09.17.558119v1?rss=1">
<title>
<![CDATA[
The diverse evolutionary histories of domesticated metaviral capsid genes in mammals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.17.558119v1?rss=1</link>
<description><![CDATA[
Selfish genetic elements and their remnants comprise at least half of the human genome. Active transposons duplicate by inserting copies at new sites in a host genome. Following insertion, transposons can acquire mutations that render them inactive; the accrual of additional mutations can render them unrecognizable over time. However, in rare instances, segments of transposons become useful for the host, in a process called gene domestication. Using the first complete human genome assembly and 25 additional vertebrate genomes, we analyzed the evolutionary trajectories and functional potential of genes domesticated from the capsid genes of Metaviridae, a retroviral-like retrotransposon family. Our analysis reveals four families of domesticated capsid genes in placental mammals with varied evolutionary outcomes, ranging from universal retention to lineage-specific duplications or losses and from purifying selection to lineage-specific rapid evolution. The four families of domesticated capsid genes have divergent amino-terminal domains, inherited from four distinct ancestral metaviruses. Structural predictions reveal that many domesticated genes encode a previously unrecognized RNA-binding domain retained in multiple paralogs in mammalian genomes both adjacent to and independent from the capsid domain. Collectively, our study reveals diverse outcomes of domestication of diverse metaviruses, which led to structurally and evolutionarily diverse genes that encode important, but still largely-unknown functions in placental mammals.
]]></description>
<dc:creator>Henriques, W. S.</dc:creator>
<dc:creator>Young, J. M.</dc:creator>
<dc:creator>Nemudryi, A.</dc:creator>
<dc:creator>Nemudraia, A.</dc:creator>
<dc:creator>Wiedenheft, B.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2023-09-17</dc:date>
<dc:identifier>doi:10.1101/2023.09.17.558119</dc:identifier>
<dc:title><![CDATA[The diverse evolutionary histories of domesticated metaviral capsid genes in mammals]]></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/10.1101/2023.09.20.558501v1?rss=1">
<title>
<![CDATA[
Alternate splice variants of the mitochondrial fission protein DNM1L/Drp1 regulate mitochondrial dynamics and cell fate in ovarian cancer. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.20.558501v1?rss=1</link>
<description><![CDATA[
Aberrant mitochondrial fission/fusion dynamics have been reported in cancer cells. While post translational modifications are known regulators of the mitochondrial fission/fusion machinery, we show that alternative splice variants of the fission protein Drp1 (DNM1L) have specific and unique roles in cancer, adding to the complexity of mitochondrial fission/fusion regulation in tumor cells. Ovarian cancer specimens express an alternative splice transcript variant of Drp1 lacking exon 16 of the variable domain, and high expression of this splice variant relative to other transcripts is associated with poor patient outcome. Unlike the full-length variant, expression of Drp1 lacking exon 16 leads to decreased association of Drp1 to mitochondrial fission sites, more fused mitochondrial networks, enhanced respiration, and TCA cycle metabolites, and is associated with a more metastatic phenotype in vitro and in vivo. These pro-tumorigenic effects can also be inhibited by specific siRNA-mediated inhibition of the endogenously expressed transcript lacking exon 16. Moreover, lack of exon 16 abrogates mitochondrial fission in response to pro-apoptotic stimuli and leads to decreased sensitivity to chemotherapeutics. These data emphasize the significance of the pathophysiological consequences of Drp1 alternative splicing and divergent functions of Drp1 splice variants, and strongly warrant consideration of Drp1 splicing in future studies.
]]></description>
<dc:creator>Javed, Z.</dc:creator>
<dc:creator>Shin, D. H.</dc:creator>
<dc:creator>Pan, W.</dc:creator>
<dc:creator>White, S. R.</dc:creator>
<dc:creator>Kim, Y. S.</dc:creator>
<dc:creator>Elhaw, A. T.</dc:creator>
<dc:creator>Kamlapurkar, S.</dc:creator>
<dc:creator>Cheng, Y.-Y.</dc:creator>
<dc:creator>Benson, J. C.</dc:creator>
<dc:creator>Abdelnaby, A. E.</dc:creator>
<dc:creator>Phaeton, R.</dc:creator>
<dc:creator>Wang, H.-G.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Sullivan, M. L. G.</dc:creator>
<dc:creator>Watkins, S. C.</dc:creator>
<dc:creator>Mullett, S. J.</dc:creator>
<dc:creator>Gelhaus, S. L.</dc:creator>
<dc:creator>Lee, N. Y.</dc:creator>
<dc:creator>Coffman, L.</dc:creator>
<dc:creator>Aird, K. M.</dc:creator>
<dc:creator>Trebak, M.</dc:creator>
<dc:creator>Karthikeyan, M.</dc:creator>
<dc:creator>Walter, V.</dc:creator>
<dc:creator>Hempel, N.</dc:creator>
<dc:date>2023-09-20</dc:date>
<dc:identifier>doi:10.1101/2023.09.20.558501</dc:identifier>
<dc:title><![CDATA[Alternate splice variants of the mitochondrial fission protein DNM1L/Drp1 regulate mitochondrial dynamics and cell fate in ovarian cancer.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.20.558720v1?rss=1">
<title>
<![CDATA[
Computational design of sequence-specific DNA-binding proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.20.558720v1?rss=1</link>
<description><![CDATA[
Sequence-specific DNA-binding proteins (DBPs) play critical roles in biology and biotechnology, and there has been considerable interest in the engineering of DBPs with new or altered specificities for genome editing and other applications. While there has been some success in reprogramming naturally occurring DBPs using selection methods, the computational design of new DBPs that recognize arbitrary target sites remains an outstanding challenge. We describe a computational method for the design of small DBPs that recognize specific target sequences through interactions with bases in the major groove, and employ this method in conjunction with experimental screening to generate binders for 5 distinct DNA targets. These binders exhibit specificity closely matching the computational models for the target DNA sequences at as many as 6 base positions and affinities as low as 30-100 nM. The crystal structure of a designed DBP-target site complex is in close agreement with the design model, highlighting the accuracy of the design method. The designed DBPs function in both Escherichia coli and mammalian cells to repress and activate transcription of neighboring genes. Our method is a substantial step towards a general route to small and hence readily deliverable sequence-specific DBPs for gene regulation and editing.
]]></description>
<dc:creator>Glasscock, C. J.</dc:creator>
<dc:creator>Pecoraro, R.</dc:creator>
<dc:creator>McHugh, R.</dc:creator>
<dc:creator>Doyle, L. A.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Boivin, O.</dc:creator>
<dc:creator>Lonnquist, B.</dc:creator>
<dc:creator>Na, E.</dc:creator>
<dc:creator>Politanska, Y.</dc:creator>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Cox, D.</dc:creator>
<dc:creator>Norn, C.</dc:creator>
<dc:creator>Coventry, B.</dc:creator>
<dc:creator>Goreshnik, I.</dc:creator>
<dc:creator>Vafeados, D.</dc:creator>
<dc:creator>Lee, G. R.</dc:creator>
<dc:creator>Gordan, R.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>DiMaio, F.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2023-09-21</dc:date>
<dc:identifier>doi:10.1101/2023.09.20.558720</dc:identifier>
<dc:title><![CDATA[Computational design of sequence-specific DNA-binding proteins]]></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/10.1101/2023.09.22.559069v1?rss=1">
<title>
<![CDATA[
SurvBal: Compositional Microbiome Balances for Survival Outcomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.22.559069v1?rss=1</link>
<description><![CDATA[
Identification of balances of bacterial taxa in relation to continuous and dichotomous outcomes is an increasingly frequent analytic objective in microbiome profiling experiments. SurvBal enables the selection of balances in relation to censored survival or time-to-event outcomes which are of considerable interest in many biomedical studies. The most commonly used survival models - the Cox proportional hazards and parametric survival models are included in the package, which are used in combination with step-wise selection procedures to identify the optimal associated balance of microbiome, i.e., the ratio of the geometric means of two groups of taxas relative abundances.

Availability and implementationSurvBal is available as an R package and a Shiny app: https://github.com/yinglia/SurvBal, https://yinglistats.shinyapps.io/shinyapp-survbal/.
]]></description>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Lee, T.</dc:creator>
<dc:creator>Marin, K.</dc:creator>
<dc:creator>Hua, X.</dc:creator>
<dc:creator>Srinivasan, S.</dc:creator>
<dc:creator>Fredricks, D. N.</dc:creator>
<dc:creator>Lee, J. R.</dc:creator>
<dc:creator>Ling, W.</dc:creator>
<dc:date>2023-09-24</dc:date>
<dc:identifier>doi:10.1101/2023.09.22.559069</dc:identifier>
<dc:title><![CDATA[SurvBal: Compositional Microbiome Balances for Survival Outcomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.25.556595v1?rss=1">
<title>
<![CDATA[
RADIF(C1orf112)-FIGNL1 Complex Regulates RAD51 Chromatin Association to Promote Viability After Replication Stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.25.556595v1?rss=1</link>
<description><![CDATA[
Homologous recombination (HR) plays critical roles in repairing lesions that arise during DNA replication and is thus essential for viability. RAD51 plays important roles during replication and HR, however, how RAD51 is regulated downstream of nucleofilament formation and how the varied RAD51 functions are regulated is not clear. We have investigated the poorly characterized protein c1orf112/RADIF that previously scored in genome-wide screens for mediators of DNA inter-strand crosslink (ICL) repair. Upon ICL agent exposure, RADIF loss leads to marked cell death, elevated chromosomal instability, increased micronuclei formation, altered cell cycle progression and increased DNA damage signaling. RADIF is recruited to damage foci and forms a complex with FIGNL1. Both proteins have epistatic roles in ICL repair, forming a co-stable complex. Mechanistically, RADIF loss leads to increased RAD51 amounts and foci on chromatin both with or without exogenous DNA damage, defective replication fork progression and reduced HR competency. We posit that RADIF is essential for limiting RAD51 levels on chromatin in the absence of damage and for RAD51 dissociation from nucleofilaments to properly complete HR. Failure to do so leads to replication slowing and inability to complete repair.
]]></description>
<dc:creator>Tischler, J. D.</dc:creator>
<dc:creator>Tsuchida, H.</dc:creator>
<dc:creator>Bosire, R.</dc:creator>
<dc:creator>Oda, T. T.</dc:creator>
<dc:creator>Park, A.</dc:creator>
<dc:creator>Adeyemi, R. O.</dc:creator>
<dc:date>2023-09-25</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.556595</dc:identifier>
<dc:title><![CDATA[RADIF(C1orf112)-FIGNL1 Complex Regulates RAD51 Chromatin Association to Promote Viability After Replication Stress]]></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/10.1101/2023.09.13.557606v1?rss=1">
<title>
<![CDATA[
The effect of single mutations in Zika virus envelope on escape from broadly neutralizing antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.13.557606v1?rss=1</link>
<description><![CDATA[
Zika virus and dengue virus are co-circulating flaviviruses with a widespread endemic range. Eliciting broad and potent neutralizing antibodies is an attractive goal for developing a vaccine to simultaneously protect against these viruses. However, the capacity of viral mutations to confer escape from broadly neutralizing antibodies remains undescribed, due in part to limited throughput and scope of traditional approaches. Here, we use deep mutational scanning to map how all possible single amino acid mutations in Zika virus envelope protein affect neutralization by antibodies of varying breadth and potency. While all antibodies selected viral escape mutations, the mutations selected by broadly neutralizing antibodies conferred less escape relative to those selected by narrow, virus-specific antibodies. Surprisingly, even for broadly neutralizing antibodies with similar binding footprints, different single mutations led to escape, indicating distinct functional requirements for neutralization not captured by existing structures. Additionally, the antigenic effects of mutations selected by broadly neutralizing antibodies were conserved across divergent, albeit related, flaviviruses. Our approach identifies residues critical for antibody neutralization, thus comprehensively defining the as-yet-unknown functional epitopes of antibodies with clinical potential.

ImportanceThe wide endemic range of mosquito-vectored flaviviruses - such as Zika virus and dengue virus serotypes 1-4 - places hundreds of millions of people at risk of infection every year. Despite this, there are no widely available vaccines, and treatment of severe cases is limited to supportive care. An avenue towards development of more widely applicable vaccines and targeted therapies is the characterization of monoclonal antibodies that broadly neutralize all these viruses. Here, we measure how single amino acid mutations in viral envelope protein affect neutralizing antibodies with both broad and narrow specificities. We find that broadly neutralizing antibodies with potential as vaccine prototypes or biological therapeutics are quantifiably more difficult to escape than narrow, virus-specific neutralizing antibodies.
]]></description>
<dc:creator>Goo, L.</dc:creator>
<dc:creator>Kikawa, C.</dc:creator>
<dc:creator>Cartwright-Acar, C. H.</dc:creator>
<dc:creator>Stuart, J. B.</dc:creator>
<dc:creator>Contreras, M.</dc:creator>
<dc:creator>Levoir, L. M.</dc:creator>
<dc:creator>Evans, M. J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2023-09-26</dc:date>
<dc:identifier>doi:10.1101/2023.09.13.557606</dc:identifier>
<dc:title><![CDATA[The effect of single mutations in Zika virus envelope on escape from broadly neutralizing antibodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.27.559793v1?rss=1">
<title>
<![CDATA[
Massively parallel identification of sequence motifs triggering ribosome-associated mRNA quality control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.27.559793v1?rss=1</link>
<description><![CDATA[
Decay of mRNAs can be triggered by ribosome slowdown at stretches of rare codons or positively charged amino acids. However, the full diversity of sequences that trigger co-translational mRNA decay is poorly understood. To comprehensively identify sequence motifs that trigger mRNA decay, we use a massively parallel reporter assay to measure the effect of all possible combinations of codon pairs on mRNA levels in S. cerevisiae. In addition to known mRNA-destabilizing sequences, we identify several dipeptide repeats whose translation reduces mRNA levels. These include combinations of positively charged and bulky residues, as well as proline-glycine and proline-aspartate dipeptide repeats. Genetic deletion of the ribosome collision sensor Hel2 rescues the mRNA effects of these motifs, suggesting that they trigger ribosome slowdown and activate the ribosome-associated quality control (RQC) pathway. Deep mutational scanning of an mRNA-destabilizing dipeptide repeat reveals a complex interplay between the charge, bulkiness, and location of amino acid residues in conferring mRNA instability. Finally, we show that the mRNA effects of codon pairs are predictive of the effects of endogenous sequences. Our work highlights the complexity of sequence motifs driving co-translational mRNA decay in eukaryotes, and presents a high throughput approach to dissect their requirements at the codon level.
]]></description>
<dc:creator>Chen, K. Y.</dc:creator>
<dc:creator>Park, H.</dc:creator>
<dc:creator>Subramaniam, A. R.</dc:creator>
<dc:date>2023-09-27</dc:date>
<dc:identifier>doi:10.1101/2023.09.27.559793</dc:identifier>
<dc:title><![CDATA[Massively parallel identification of sequence motifs triggering ribosome-associated mRNA quality control]]></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/10.1101/2023.09.30.560315v1?rss=1">
<title>
<![CDATA[
Forward genetic screens identify mechanisms of resistance to small molecule lactate dehydrogenase inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.30.560315v1?rss=1</link>
<description><![CDATA[
Altered metabolism is a hallmark of cancer; however, it has been difficult to specifically target metabolism in cancer for therapeutic benefit. Cancers with genetically defined defects in metabolic enzymes constitute a subset of cancers where targeting metabolism is potentially accessible. Hurthle cell carcinoma of the thyroid (HTC) tumors frequently harbor deleterious mitochondrial DNA (mtDNA) mutations in subunits of complex I of the mitochondrial electron transport chain (ETC). Previous work has shown that HTC models with deleterious mtDNA mutations exhibit mitochondrial ETC defects that expose lactate dehydrogenase (LDH) as a therapeutic vulnerability. Here, we performed forward genetic screens to identify mechanisms of resistance to small molecule LDH inhibitors. We identified two distinct mechanisms of resistance: upregulation of an LDH isoform and a compound-specific resistance mutation. Using these tools, we demonstrate that the anti-cancer activity of LDH inhibitors in cell line and xenograft models of complex I-mutant HTC is through on-target LDH inhibition.
]]></description>
<dc:creator>Frank, A. R.</dc:creator>
<dc:creator>Vandiver, F.</dc:creator>
<dc:creator>McFadden, D. G.</dc:creator>
<dc:date>2023-09-30</dc:date>
<dc:identifier>doi:10.1101/2023.09.30.560315</dc:identifier>
<dc:title><![CDATA[Forward genetic screens identify mechanisms of resistance to small molecule lactate dehydrogenase inhibitors]]></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/10.1101/2023.09.28.560025v1?rss=1">
<title>
<![CDATA[
ONECUT2 Activates Diverse Resistance Drivers of Androgen Receptor-Independent Heterogeneity in Prostate Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.28.560025v1?rss=1</link>
<description><![CDATA[
Significance StatementONECUT2 (OC2) is a master transcription factor that alters lineage identity by activating gene networks associated with both neuroendocrine prostate cancer and prostate adenocarcinoma. A small molecule inhibitor of OC2 represses the lineage plasticity program activated by enzalutamide, suggesting OC2 inhibition as a novel therapeutic strategy to prevent emergence of treatment-resistant variants.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=135 SRC="FIGDIR/small/560025v2_ufig1.gif" ALT="Figure 1">
View larger version (41K):
org.highwire.dtl.DTLVardef@13278a4org.highwire.dtl.DTLVardef@8e82aeorg.highwire.dtl.DTLVardef@25a115org.highwire.dtl.DTLVardef@728eb4_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOGraphic AbstractC_FLOATNO C_FIG Androgen receptor-(AR-) indifference is a mechanism of resistance to hormonal therapy in prostate cancer (PC). Here we demonstrate that the HOX/CUT transcription factor ONECUT2 (OC2) activates resistance through multiple drivers associated with adenocarcinoma, stem-like and neuroendocrine (NE) variants. Direct OC2 targets include the glucocorticoid receptor and the NE splicing factor SRRM4, among others. OC2 regulates gene expression by promoter binding, enhancement of chromatin accessibility, and formation of novel super-enhancers. OC2 also activates glucuronidation genes that irreversibly disable androgen, thereby evoking phenotypic heterogeneity indirectly by hormone depletion. Pharmacologic inhibition of OC2 suppresses lineage plasticity reprogramming induced by the AR signaling inhibitor enzalutamide. These results demonstrate that OC2 activation promotes a range of drug resistance mechanisms associated with treatment-emergent lineage variation in PC. Our findings support enhanced efforts to therapeutically target this protein as a means of suppressing treatment-resistant disease.
]]></description>
<dc:creator>Qian, C.</dc:creator>
<dc:creator>Yang, Q.</dc:creator>
<dc:creator>Rotinen, M.</dc:creator>
<dc:creator>Huang, R.</dc:creator>
<dc:creator>Kim, h.</dc:creator>
<dc:creator>Gallent, B.</dc:creator>
<dc:creator>Yan, Y.</dc:creator>
<dc:creator>Cadaneanu, R. M.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Kaochar, S.</dc:creator>
<dc:creator>Freedland, S. J.</dc:creator>
<dc:creator>Posadas, E. M.</dc:creator>
<dc:creator>Ellis, L.</dc:creator>
<dc:creator>Di Vizio, D.</dc:creator>
<dc:creator>Morrissey, C.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Brady, L.</dc:creator>
<dc:creator>Murali, R.</dc:creator>
<dc:creator>Campbell, M. J.</dc:creator>
<dc:creator>Yang, W.</dc:creator>
<dc:creator>Knudsen, B. S.</dc:creator>
<dc:creator>Mostaghel, E. A.</dc:creator>
<dc:creator>Ye, H.</dc:creator>
<dc:creator>Garraway, I. P.</dc:creator>
<dc:creator>You, S.</dc:creator>
<dc:creator>Freeman, M. R.</dc:creator>
<dc:date>2023-10-01</dc:date>
<dc:identifier>doi:10.1101/2023.09.28.560025</dc:identifier>
<dc:title><![CDATA[ONECUT2 Activates Diverse Resistance Drivers of Androgen Receptor-Independent Heterogeneity in Prostate Cancer]]></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/10.1101/2023.09.30.560314v1?rss=1">
<title>
<![CDATA[
ATF7IP2/MCAF2 directs H3K9 methylation and meiotic gene regulation in the male germline 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.30.560314v1?rss=1</link>
<description><![CDATA[
H3K9 tri-methylation (H3K9me3) plays emerging roles in gene regulation, beyond its accumulation on pericentric constitutive heterochromatin. It remains a mystery why and how H3K9me3 undergoes dynamic regulation in male meiosis. Here, we identify a novel, critical regulator of H3K9 methylation and spermatogenic heterochromatin organization: the germline-specific protein ATF7IP2 (MCAF2). We show that, in male meiosis, ATF7IP2 amasses on autosomal and X pericentric heterochromatin, spreads through the entirety of the sex chromosomes, and accumulates on thousands of autosomal promoters and retrotransposon loci. On the sex chromosomes, which undergo meiotic sex chromosome inactivation (MSCI), the DNA damage response pathway recruits ATF7IP2 to X pericentric heterochromatin, where it facilitates the recruitment of SETDB1, a histone methyltransferase that catalyzes H3K9me3. In the absence of ATF7IP2, male germ cells are arrested in meiotic prophase I. Analyses of ATF7IP2-deficient meiosis reveal the proteins essential roles in the maintenance of MSCI, suppression of retrotransposons, and global upregulation of autosomal genes. We propose that ATF7IP2 is a downstream effector of the DDR pathway in meiosis that coordinates the organization of heterochromatin and gene regulation through the spatial regulation of SETDB1-mediated H3K9me3 deposition.
]]></description>
<dc:creator>Alavattam, K. G.</dc:creator>
<dc:creator>Esparza, J. M.</dc:creator>
<dc:creator>Hu, M.</dc:creator>
<dc:creator>Shimada, R.</dc:creator>
<dc:creator>Kohrs, A. R.</dc:creator>
<dc:creator>Abe, H.</dc:creator>
<dc:creator>Munakata, Y.</dc:creator>
<dc:creator>Otsuka, K.</dc:creator>
<dc:creator>Yoshimura, S.</dc:creator>
<dc:creator>Kitamura, Y.</dc:creator>
<dc:creator>Yeh, Y.-H.</dc:creator>
<dc:creator>Hu, Y.-C.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Andreassen, P. R.</dc:creator>
<dc:creator>Ishiguro, K.-i.</dc:creator>
<dc:creator>Namekawa, S. H.</dc:creator>
<dc:date>2023-10-02</dc:date>
<dc:identifier>doi:10.1101/2023.09.30.560314</dc:identifier>
<dc:title><![CDATA[ATF7IP2/MCAF2 directs H3K9 methylation and meiotic gene regulation in the male germline]]></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/10.1101/2023.10.01.560227v1?rss=1">
<title>
<![CDATA[
Arrested Agonist Paradigm For Selective Radiosensitization of Prostate Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.01.560227v1?rss=1</link>
<description><![CDATA[
As a prototypical nuclear hormone receptor, the androgen receptor (AR) signals via a sequential cascade triggered by binding to androgenic ligands such as testosterone and dihydrotestosterone (DHT). This cascade includes dimerization of the ligand-receptor complex, nuclear translocation, chromatin binding to response elements, recruitment of TOP2B and co-activator complexes, and induction of an effector transcriptional program. In prostate cancers, this AR signaling cascade is an essential driver of growth and survival, yet its activity confers potential vulnerabilities through transient TOP2B-mediated DNA double strand breaks. We investigated the ability of non-steroidal AR ligands to activate initial steps of the AR signaling cascade up to the point of AR- and TOP2B-mediated double strand breaks, with subsequent arrest of the signaling cascade to prevent induction of pro-growth/survival transcriptional programs in prostate cancer cells. We identified hydroxyflutamide (FLU) as such an androgen receptor arrested agonist; in androgen-deprived conditions, FLU induced AR nuclear translocation, chromatin binding, and TOP2B-mediated double strand breaks, but failed to induce AR target gene expression and prostate cancer cell growth. The FLU-mediated arrest in the signaling cascade could be attributed to the inability of FLU to allow association of AR with SMARCD2, a critical component of the BAF chromatin remodeling complex required for androgen induced AR co-activation. Interestingly, the FLU-induced, AR- and TOP2B-mediated double strand breaks could be used to selectively sensitize AR-positive prostate cancer cells to ionizing radiation in vitro and in vivo. These findings support a novel arrested agonist paradigm for selective radiosensitization of prostate cancer cells without inducing AR-mediated pro-growth and survival transcriptional programs.
]]></description>
<dc:creator>Coulter, J. B.</dc:creator>
<dc:creator>Haffner, M.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Zhou, H.</dc:creator>
<dc:creator>Pham, M.-T. N.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Chikarmane, R.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:creator>Mehl, M. S.</dc:creator>
<dc:creator>Kazibwe, S.</dc:creator>
<dc:creator>Choi, K.</dc:creator>
<dc:creator>Archey, A.</dc:creator>
<dc:creator>Valluri, S.</dc:creator>
<dc:creator>Lupold, S. E.</dc:creator>
<dc:creator>Song, D.</dc:creator>
<dc:creator>De Marzo, A. M.</dc:creator>
<dc:creator>Nelson, W. G.</dc:creator>
<dc:creator>DeWeese, T. L.</dc:creator>
<dc:creator>Yegnasubramanian, S.</dc:creator>
<dc:date>2023-10-02</dc:date>
<dc:identifier>doi:10.1101/2023.10.01.560227</dc:identifier>
<dc:title><![CDATA[Arrested Agonist Paradigm For Selective Radiosensitization of Prostate Cancer]]></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/10.1101/2023.10.03.560752v1?rss=1">
<title>
<![CDATA[
Therapeutic Targeting of TIM-4-L With Engineered T Cells for Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.03.560752v1?rss=1</link>
<description><![CDATA[
Disruption of the lipid asymmetric bilayer is a common feature observed in cancer cells. We utilized the natural immune receptor TIM-4 to interrogate for loss of plasma membrane phospholipid polarity in primary acute myelogenous leukemia (AML) samples. We performed FACs analysis in 33 patients and correlated TIM-4-L expression frequency and intensity with molecular disease characteristics. In normal tissues, TIM-4-L is confined to the internal leaflet of the plasma membrane. By contrast, 86% of untreated AML blasts in our analysis displayed upregulation of cell surface TIM-4-L. These observations were agnostic to AML genetic classification, as samples with mutations in TP53, ASXL1, and RUNX1, also displayed TIM-4-L upregulation similar to that seen in favorable and intermediate subtypes. This TIM-4-L dysregulation was also stably present in both Kasumi-1 and MV-4-11 AML cell lines. To evaluate the potential of upregulated TIM-4-L to serve as a target for adoptive T cell therapy (ACT), we constructed TIM-4-L-directed engineered T cells, which demonstrated potent anti-leukemic effects, effectively eliminating AML cell lines both in vitro and in vivo. This approach led to the eradication of AML cells across a range of endogenous TIM-4-L expression levels. These results highlight TIM-4-L as a highly prevalent and novel target for T cell-based therapy in AML. Further investigations into the role of TIM-4-L in AML pathogenesis and its potential as an anti-leukemic target for clinical development are warranted.
]]></description>
<dc:creator>Cieniewicz, B.</dc:creator>
<dc:creator>Oliveira, E.</dc:creator>
<dc:creator>Saxton, M.</dc:creator>
<dc:creator>Torabi, D.</dc:creator>
<dc:creator>Bhatta, A.</dc:creator>
<dc:creator>Kukutla, P.</dc:creator>
<dc:creator>Arballo, A.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Fate, M.</dc:creator>
<dc:creator>Ning, H.</dc:creator>
<dc:creator>Corey, L.</dc:creator>
<dc:creator>Maiti, A.</dc:creator>
<dc:creator>Corey, D.</dc:creator>
<dc:date>2023-10-04</dc:date>
<dc:identifier>doi:10.1101/2023.10.03.560752</dc:identifier>
<dc:title><![CDATA[Therapeutic Targeting of TIM-4-L With Engineered T Cells for Acute Myeloid Leukemia]]></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/10.1101/2023.10.04.560669v1?rss=1">
<title>
<![CDATA[
Natural malaria infection elicits rare but potent neutralizing antibodies to the blood-stage antigen RH5 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.04.560669v1?rss=1</link>
<description><![CDATA[
Plasmodium falciparum RH5 is the most advanced blood-stage malaria vaccine candidate and is under evaluation for efficacy in endemic regions, emphasizing the need to study the underlying antibody response to RH5 during natural infection. Here, we found that RH5-reactive B cells were rare in malaria-exposed individuals despite repeated infections over multiple years. RH5-specific monoclonal antibodies isolated from these individuals were extensively mutated but mostly targeted non-neutralizing epitopes, in contrast to antibodies from RH5-vaccinated, malaria-naive individuals. However, infection-derived MAD8-151 and MAD8-502 were among the most potent neutralizers out of 186 antibodies isolated from both cohorts and target the same epitopes as the most effective vaccine-induced antibodies. Binding to basigin receptor-proximal epitopes was the primary factor governing the potency of RH5-specific antibodies from both natural infection and vaccination, followed by the strength of binding. These results indicate a clear strategy for the development of next-generation RH5 vaccines for use in malaria-endemic regions.
]]></description>
<dc:creator>Wang, L. T.</dc:creator>
<dc:creator>Cooper, A. J. R.</dc:creator>
<dc:creator>Farrell, B.</dc:creator>
<dc:creator>Miura, K.</dc:creator>
<dc:creator>Diouf, A.</dc:creator>
<dc:creator>Muller-Sienerth, N.</dc:creator>
<dc:creator>Crosnier, C.</dc:creator>
<dc:creator>Purser, L.</dc:creator>
<dc:creator>Maciuszek, M.</dc:creator>
<dc:creator>Barrett, J. R.</dc:creator>
<dc:creator>McHugh, K.</dc:creator>
<dc:creator>Tucker, C.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Doumbo, S.</dc:creator>
<dc:creator>Doumtabe, D.</dc:creator>
<dc:creator>Pyo, C.-W.</dc:creator>
<dc:creator>Nielsen, C. M.</dc:creator>
<dc:creator>Silk, S. E.</dc:creator>
<dc:creator>Kayentao, K.</dc:creator>
<dc:creator>Ongoiba, A.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Nguyen, D. C.</dc:creator>
<dc:creator>Lee, F. E.-H.</dc:creator>
<dc:creator>Minassian, A. M.</dc:creator>
<dc:creator>Geraghty, D. E.</dc:creator>
<dc:creator>Traore, B.</dc:creator>
<dc:creator>Seder, R. A.</dc:creator>
<dc:creator>Crompton, P. D.</dc:creator>
<dc:creator>Wright, G. J.</dc:creator>
<dc:creator>Long, C. A.</dc:creator>
<dc:creator>Draper, S. J.</dc:creator>
<dc:creator>Higgins, M. K.</dc:creator>
<dc:creator>Tan, J.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.04.560669</dc:identifier>
<dc:title><![CDATA[Natural malaria infection elicits rare but potent neutralizing antibodies to the blood-stage antigen RH5]]></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/10.1101/2023.10.06.561250v1?rss=1">
<title>
<![CDATA[
Deep mutational scanning of CYP2C19 reveals a substrate specificity-abundance tradeoff 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.06.561250v1?rss=1</link>
<description><![CDATA[
Cytochrome P450s (CYPs) are a family of enzymes responsible for metabolizing nearly 80% of small molecule drugs. Variants in CYPs can substantially alter drug metabolism, which may result in improper dosing and severe adverse drug reactions. CYPs have low sequence conservation, making it difficult to anticipate whether variant effects measured in one CYP may extend to others based on sequence alone. Even closely related CYPs, like CYP2C9 and its closest homolog CYP2C19, have distinct phenotypic properties despite sharing 92% amino acid sequence identity. Thus, we used Variant Abundance by Massively Parallel sequencing (VAMP-seq) to measure the steady-state protein abundance, a proxy for protein stability, of 7,660 missense variants in CYP2C19 expressed in cultured human cells. Our results confirmed positions and structural features critical for CYP function and revealed how variants at positions conserved across all eukaryotic CYPs influence abundance. We jointly analyzed 4,670 variants whose abundance was measured in both CYP2C19 and CYP2C9, finding that the homologs have different variant abundances in substrate recognition sites within the hydrophobic core, and that substitutions in some regions reduced abundance in CYP2C19 but not CYP2C9. We also measured the abundance of all single and some multiple WT amino acid exchanges between CYP2C19 and CYP2C9. While most exchanges had no effect, substitutions in substrate recognition site 4 (SRS4) reduced abundance in CYP2C19. When nearby amino acids were exchanged in double and triple mutants, we found distinct interactions between the sites in CYP2C19 and CYP2C9, revealing a region that is partially responsible for the difference in thermodynamic stability between the two homologs. Since these positions are also important for determining substrate specificity, there may be an evolutionary tradeoff between stability and altered enzymatic function. Finally, we used our data to analyze 368 previously unannotated human variants, finding that 43% had decreased abundance. Thus, by comparing variant effects between two closely related and important human genes, we have uncovered regions underlying their functional differences and paved the way for a more complete understanding of one of the most versatile families of enzymes.
]]></description>
<dc:creator>Boyle, G. E.</dc:creator>
<dc:creator>Sitko, K.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Bianchi, A. H.</dc:creator>
<dc:creator>Dixon, A.</dc:creator>
<dc:creator>Thomson, R. E. S.</dc:creator>
<dc:creator>Garge, R. K.</dc:creator>
<dc:creator>Rettie, A. E.</dc:creator>
<dc:creator>Rubin, A. F.</dc:creator>
<dc:creator>Geck, R. C.</dc:creator>
<dc:creator>Gillam, E. M. J.</dc:creator>
<dc:creator>DeWitt, W. S.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Fowler, D. M.</dc:creator>
<dc:date>2023-10-08</dc:date>
<dc:identifier>doi:10.1101/2023.10.06.561250</dc:identifier>
<dc:title><![CDATA[Deep mutational scanning of CYP2C19 reveals a substrate specificity-abundance tradeoff]]></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/10.1101/2023.10.13.562275v1?rss=1">
<title>
<![CDATA[
Antigen-level resolution of commensal-specific B cell responses enabled by phage-display screening and B cell tetramers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.13.562275v1?rss=1</link>
<description><![CDATA[
Induction of adaptive immune responses to commensal microbes is critical for tissue homeostasis, and perturbation of these responses is associated with multiple chronic inflammatory disorders. However, the mechanisms underlying the induction and regulation of mucosal B cells targeting commensal microbes remain poorly understood, in part due to a lack of tools to identify commensal-specific B cells ex vivo. To address this, we identified immunogenic protein epitopes recognized by Segmented Filamentous Bacteria (SFB)-specific serum antibodies using a whole-genome phage display screen and identified immunogenic proteins engaging IgA, IgG1 and IgG2b responses. Using these antigens, we generated B cell tetramers to identify and track SFB-specific B cell responses in the gut associated lymphoid tissue during natural and de novo colonization. We revealed a compartmentalized response in SFB-specific B cell activation between Peyers patches and mesenteric lymph nodes, with a gradient of IgA, IgG1 and IgG2b isotypes along the small intestine, and selective production of IgG2b with the mesenteric lymph node chain. VDJ sequencing analyses and generation of SFB-specific monoclonal antibodies identified that somatic hypermutation drives affinity maturation to SFB derived antigens under homeostatic conditions. By combining phage display screening and B cell tetramer technologies, we now enable antigen-level based studies of immunity to intestinal microbes, which will advance our understanding of the ontogeny and function of commensal-specific B cell responses in tissue immunity, inflammation and repair.
]]></description>
<dc:creator>Verma, S.</dc:creator>
<dc:creator>Dufort, M. J.</dc:creator>
<dc:creator>Olsen, T. M.</dc:creator>
<dc:creator>Labuda, J. C.</dc:creator>
<dc:creator>Kimmel, S.</dc:creator>
<dc:creator>Scharffenberger, S.</dc:creator>
<dc:creator>McGuire, A.</dc:creator>
<dc:creator>Harrison, O. J.</dc:creator>
<dc:date>2023-10-17</dc:date>
<dc:identifier>doi:10.1101/2023.10.13.562275</dc:identifier>
<dc:title><![CDATA[Antigen-level resolution of commensal-specific B cell responses enabled by phage-display screening and B cell tetramers]]></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/10.1101/2023.10.17.562709v1?rss=1">
<title>
<![CDATA[
A novel approach to exploring the dark genome and its application to mapping of the vertebrate virus 'fossil record'. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.17.562709v1?rss=1</link>
<description><![CDATA[
BackgroundGenomic regions that remain poorly understood, often referred to as the "dark genome," contain a variety of functionally relevant and biologically informative genome features. These include endogenous viral elements (EVEs) - virus-derived sequences that can dramatically impact host biology and serve as a virus "fossil record". In this study, we introduce a database-integrated genome screening (DIGS) approach to investigating the dark genome in silico, focusing on EVEs found within vertebrate genomes.

ResultsUsing DIGS on 874 vertebrate species genomes, we uncovered approximately 1.1 million EVE sequences, with over 99% originating from endogenous retroviruses or transposable elements that contain EVE DNA. We show that the remaining 6038 sequences represent over a thousand distinct horizontal gene transfer events across ten virus families, including some that have not previously been reported as EVEs. We explore the genomic and phylogenetic characteristics of non-retroviral EVEs and determine their rates of acquisition during vertebrate evolution. Our study uncovers novel virus diversity, broadens knowledge of virus distribution among vertebrate hosts, and provides new insights into the ecology and evolution of vertebrate viruses.

ConclusionsWe comprehensively catalogue and analyse EVEs within 874 vertebrate genomes, shedding light on the distribution, diversity and long-term evolution of viruses, and revealing their extensive impact on vertebrate genome evolution. Our results demonstrate the power of linking a relational database management system to a similarity search-based screening pipeline for in silico exploration of the dark genome.
]]></description>
<dc:creator>Blanco-Melo, D.</dc:creator>
<dc:creator>Campbell, M. A.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Dennis, T.</dc:creator>
<dc:creator>Mohda, S.</dc:creator>
<dc:creator>Lytras, S.</dc:creator>
<dc:creator>Hughes, J. J.</dc:creator>
<dc:creator>Gatseva, A.</dc:creator>
<dc:creator>Gifford, R. J.</dc:creator>
<dc:date>2023-10-19</dc:date>
<dc:identifier>doi:10.1101/2023.10.17.562709</dc:identifier>
<dc:title><![CDATA[A novel approach to exploring the dark genome and its application to mapping of the vertebrate virus 'fossil record'.]]></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/10.1101/2023.10.20.563295v1?rss=1">
<title>
<![CDATA[
Terpenoid balance in Aspergillus nidulans unveiled by heterologous squalene synthase expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.20.563295v1?rss=1</link>
<description><![CDATA[
Filamentous fungi produce numerous uncharacterized natural products (NPs) that are often challenging to characterize due to cryptic expression in laboratory conditions. Previously, we have successfully isolated novel NPs by expressing fungal artificial chromosomes (FACs) from a variety of fungal species into Aspergillus nidulans. Here, we demonstrate a new twist to FAC utility wherein heterologous expression of a Pseudogymnoascus destructans FAC in A. nidulans altered endogenous terpene biosynthetic pathways. In contrast to wildtype, the FAC transformant produced increased levels of squalene and aspernidine type compounds, including three new nidulenes (1-2, 5), and lost nearly all ability to synthesize the major A. nidulans characteristic terpene, austinol. Deletion of a squalene synthase gene in the FAC restored wildtype chemical profiles. The altered squalene to farnesyl pyrophosphate ratio leading to synthesis of nidulenes and aspernidines at the expense of farnesyl pyrophosphate derived austinols provides unexpected insight into routes of terpene synthesis in fungi.

TeaserReshaping terpenes: Heterologous FAC expression reroutes terpene pathways.
]]></description>
<dc:creator>Park, S. C.</dc:creator>
<dc:creator>Steffan, B. N.</dc:creator>
<dc:creator>Lim, F. Y.</dc:creator>
<dc:creator>Gupta, R.</dc:creator>
<dc:creator>Butun, F. A.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Ye, R.</dc:creator>
<dc:creator>Decker, T.</dc:creator>
<dc:creator>Wu, C. C.</dc:creator>
<dc:creator>Kelleher, N. L.</dc:creator>
<dc:creator>Bok, J. W.</dc:creator>
<dc:creator>Keller, N. P.</dc:creator>
<dc:date>2023-10-20</dc:date>
<dc:identifier>doi:10.1101/2023.10.20.563295</dc:identifier>
<dc:title><![CDATA[Terpenoid balance in Aspergillus nidulans unveiled by heterologous squalene synthase expression]]></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/10.1101/2023.10.21.562237v1?rss=1">
<title>
<![CDATA[
Genotype prediction of 336,463 samples from public expression data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.21.562237v1?rss=1</link>
<description><![CDATA[
Tens of thousands of RNA-sequencing experiments comprising hundreds of thousands of individual samples have now been performed. These data represent a broad range of experimental conditions, sequencing technologies, and hypotheses under study. The Recount project has aggregated and uniformly processed hundreds of thousands of publicly available RNA-seq samples. Most of these samples only include RNA expression measurements; genotype data for these same samples would enable a wide range of analyses including variant prioritization, eQTL analysis, and studies of allele specific expression. Here, we developed a statistical model based on the existing reference and alternative read counts from the RNA-seq experiments available through Recount3 to predict genotypes at autosomal biallelic loci in coding regions. We demonstrate the accuracy of our model using large-scale studies that measured both gene expression and genotype genome-wide. We show that our predictive model is highly accurate with 99.5% overall accuracy, 99.6% major allele accuracy, and 90.4% minor allele accuracy. Our model is robust to tissue and study effects, provided the coverage is high enough. We applied this model to genotype all the samples in Recount3 and provide the largest ready-to-use expression repository containing genotype information. We illustrate that the predicted genotype from RNA-seq data is sufficient to unravel the underlying population structure of samples in Recount3 using Principal Component Analysis.
]]></description>
<dc:creator>Razi, A.</dc:creator>
<dc:creator>Lo, C. C.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Leek, J. T.</dc:creator>
<dc:creator>Hansen, K. D.</dc:creator>
<dc:date>2023-10-22</dc:date>
<dc:identifier>doi:10.1101/2023.10.21.562237</dc:identifier>
<dc:title><![CDATA[Genotype prediction of 336,463 samples from public expression data]]></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/10.1101/2023.10.23.563604v1?rss=1">
<title>
<![CDATA[
The Ribosome Assembly Factor Reh1 is Released from the Polypeptide Exit Tunnel in the Pioneer Round of Translation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.23.563604v1?rss=1</link>
<description><![CDATA[
Assembly of functional ribosomal subunits and successfully delivering them to the translating pool is a prerequisite for protein synthesis and cell growth. In S. cerevisiae, the ribosome assembly factor Reh1 binds to pre-60S subunits at a late stage during their cytoplasmic maturation. Previous work shows that the C-terminus of Reh1 inserts into the polypeptide exit tunnel (PET) of the pre-60S subunit. Unlike canonical assembly factors, which associate exclusively with pre-60S subunits, we observed that Reh1 sediments with polysomes in addition to free 60S subunits. We therefore investigated the intriguing possibility that Reh1 remains associated with 60S subunits after the release of the anti-association factor Tif6 and after subunit joining. Here, we show that Reh1-bound nascent 60S subunits associate with 40S subunits to form actively translating ribosomes. Using selective ribosome profiling, we found that Reh1-bound ribosomes populate open reading frames near start codons. Reh1-bound ribosomes are also strongly enriched for initiator tRNA, indicating they are associated with early elongation events. Using single particle cryo-electron microscopy to image cycloheximide-arrested Reh1-bound 80S ribosomes, we found that Reh1-bound 80S contain A site peptidyl tRNA, P site tRNA and eIF5A indicating that Reh1 does not dissociate from 60S until early stages of translation elongation. We propose that Reh1 is displaced by the elongating peptide chain. These results identify Reh1 as the last assembly factor released from the nascent 60S subunit during its pioneer round of translation.
]]></description>
<dc:creator>Musalgaonkar, S.</dc:creator>
<dc:creator>Yelland, J.</dc:creator>
<dc:creator>Chitale, R.</dc:creator>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Ozadam, H.</dc:creator>
<dc:creator>Cenik, C.</dc:creator>
<dc:creator>Taylor, D.</dc:creator>
<dc:creator>Johnson, A. W.</dc:creator>
<dc:date>2023-10-23</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563604</dc:identifier>
<dc:title><![CDATA[The Ribosome Assembly Factor Reh1 is Released from the Polypeptide Exit Tunnel in the Pioneer Round of Translation]]></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/10.1101/2023.10.20.563324v1?rss=1">
<title>
<![CDATA[
Germline novelty through recurrent copy-number, protein, and regulatory evolution of the synaptonemal complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.20.563324v1?rss=1</link>
<description><![CDATA[
The synaptonemal complex (SC) is a protein-rich structure essential for meiotic recombination and faithful chromosome segregation. Acting like a zipper to paired chromosomes during early prophase, the complex consists of central elements bilaterally tethered by the transverse filaments to the lateral elements anchored on either side to the homologous chromosome axes. Despite being found in most major eukaryotic taxa implying a deeply conserved evolutionary origin, several components of the complex exhibit unusually high rates of sequence turnover. This is puzzlingly exemplified by the SC of Drosophila, where the central elements and transverse filaments display no identifiable homologs outside of the genus. Here, we exhaustively examine the evolutionary history of the SC in Drosophila taking a comparative phylogenomic approach with high species density to circumvent obscured homology due to rapid sequence evolution. Contrasting starkly against other genes involved in meiotic chromosome pairing, SC significantly shows elevated rates of coding evolution due to a combination of relaxed constraint and recurrent, widespread positive selection. In particular, the central element cona and transverse filament c(3)G have diversified through tandem and retro-duplications, repeatedly generating paralogs that likely have novel germline functions. In a striking case of molecular convergence, c(3)G paralogs that independently arose in distant lineages evolved under positive selection to have convergent truncations to the protein termini and elevated testes expression. Surprisingly, the expression of SC genes in the germline is exceedingly prone to change suggesting recurrent regulatory evolution which, in many species, resulted in high testes expression even though Drosophila males are achiasmic. Overall, our study recapitulates the poor conservation of SC components, and further uncovers that the lack of conservation extends to other modalities including copy number, genomic locale, and germline regulation. Considering the elevated testes expression in many Drosophila species and the common ancestor, we suggest that the function of SC genes in the male germline, while still poorly understood, may be a prime target of constant evolutionary pressures driving repeated adaptations and innovations.

SummaryThe synaptonemal complex (SC) is essential for meiotic recombination and faithful chromosome segregation across eukaryotes, yet components of the SC are often poorly conserved. Here we show that across the Drosophila phylogeny several SC genes have evolved under recurrent positive selection resulting in orthologs that are barely recognizable. This is partly driven duplications repeatedly generating paralogs that may have adopted novel germline functions, often in the testes. Unexpectedly, while most SC genes are thought to be dispensable in the male germline where recombination is absent in Drosophila, elevated testes expression appears to be the norm across the genus and likely the ancestral state. The evolutionary lability of SC genes in Drosophila is likely a repeated source of adaptive innovations in the germline.
]]></description>
<dc:creator>Wei, K. H.</dc:creator>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Chatla, K.</dc:creator>
<dc:creator>Krishnapura, A.</dc:creator>
<dc:creator>Appiah, S. P.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Unckless, R. L.</dc:creator>
<dc:creator>Blumenstiel, J.</dc:creator>
<dc:creator>Bachtrog, D.</dc:creator>
<dc:date>2023-10-24</dc:date>
<dc:identifier>doi:10.1101/2023.10.20.563324</dc:identifier>
<dc:title><![CDATA[Germline novelty through recurrent copy-number, protein, and regulatory evolution of the synaptonemal complex]]></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/10.1101/2023.10.24.563725v1?rss=1">
<title>
<![CDATA[
A quantitative genetics framework for understanding the selection response of microbial communities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.24.563725v1?rss=1</link>
<description><![CDATA[
Heritability, a quantity that reflects the degree of resemblance between parent and offspring traits, is measured during plant and animal breeding because it predicts selection success during artificial selection of individuals. However, when whole microbial communities are under artificial selection to improve their traits, high heritability of the community trait does not necessarily predict selection success. To better understand the relationship between heritability and success during community selection, we establish a quantitative genetics framework, and in doing so, we obtain practical recommendations. Specifically, we decompose a community trait into "trait determinants": genotype compositions and species compositions that impact the community trait and that vary among communities. This allows us to interpret heritability of a community trait in terms of the heritability of its determinants. We then use the Price equation to partition the selection response of a community trait into three phenomena: inter-community selection (heritability multiplied by selection intensity), transmission infidelity (the change in community trait from parent to offspring), and nonlinearity (due to a nonlinear relationship between parent and offspring traits). We illustrate that evolution within a community can cause the three terms to covary: in addition to the known effect of worsening transmission infidelity, intra-community evolution can lead to inflated heritability values greater than one (through an effect whereby "the poor get poorer"), and simultaneously magnify nonlinearity. As a consequence of these effects, heritability no longer predicts the selection response of a community trait. We propose effective selection strategies that improve heritability without accelerating intra-community evolution.
]]></description>
<dc:creator>Xie, L.</dc:creator>
<dc:creator>Yuan, A. E.</dc:creator>
<dc:creator>Shou, W.</dc:creator>
<dc:date>2023-10-24</dc:date>
<dc:identifier>doi:10.1101/2023.10.24.563725</dc:identifier>
<dc:title><![CDATA[A quantitative genetics framework for understanding the selection response of microbial communities]]></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/10.1101/2023.10.23.563660v1?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/10.1101/2023.10.25.563990v1?rss=1">
<title>
<![CDATA[
Widespread fungal-bacterial competition for magnesium enhances antibiotic resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.25.563990v1?rss=1</link>
<description><![CDATA[
Fungi and bacteria coexist in many polymicrobial communities, yet the molecular basis of their interactions remains poorly understood. Using unbiased genomic approaches, we discover that the fungus Candida albicans sequesters essential Mg2+ ions from the bacterium Pseudomonas aeruginosa. In turn, the bacterium competes using a Mg2+ transporter, MgtA. We show that Mg2+ sequestration by fungi is a general mechanism of antagonism against gram-negative bacteria. But the resultant Mg2+ limitation enhances bacterial resistance to polymyxin antibiotics like colistin, which target gram-negative bacterial membranes. Experimental evolution reveals that bacteria in co-culture with fungi become phenotypically, but not genetically, resistant to colistin; antifungal treatment renders resistant bacteria from co-cultures to become colistin-sensitive. Fungal-bacterial nutritional competition may thus profoundly impact treatments of polymicrobial infections with antibiotics of last resort.

One Sentence SummaryMagnesium sequestration by fungi lowers bacterial fitness but enhances antibiotic resistance.
]]></description>
<dc:creator>Hsieh, Y.-Y. P.</dc:creator>
<dc:creator>Sun, W. W.</dc:creator>
<dc:creator>Young, J. M.</dc:creator>
<dc:creator>Cheung, R.</dc:creator>
<dc:creator>Hogan, D. A.</dc:creator>
<dc:creator>Dandekar, A. A.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2023-10-26</dc:date>
<dc:identifier>doi:10.1101/2023.10.25.563990</dc:identifier>
<dc:title><![CDATA[Widespread fungal-bacterial competition for magnesium enhances antibiotic resistance]]></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/10.1101/2023.10.29.564578v1?rss=1">
<title>
<![CDATA[
dms-viz: Structure-informed visualizations for deep mutational scanning and other mutation-based datasets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.29.564578v1?rss=1</link>
<description><![CDATA[
Summary and PurposeUnderstanding how mutations impact a proteins functions is valuable for many types of biological questions. High-throughput techniques such as deep-mutational scanning (DMS) have greatly expanded the number of mutation-function datasets. For instance, DMS has been used to determine how mutations to viral proteins affect antibody escape (Dadonaite et al. 2023), receptor affinity (Starr et al. 2020), and essential functions such as viral genome transcription and replication (Li et al. 2023). With the growth of sequence databases, in some cases the effects of mutations can also be inferred from phylogenies of natural sequences (Bloom and Neher 2023) (Figure 1).

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=147 SRC="FIGDIR/small/564578v1_fig1.gif" ALT="Figure 1">
View larger version (31K):
org.highwire.dtl.DTLVardef@6c623forg.highwire.dtl.DTLVardef@1a62199org.highwire.dtl.DTLVardef@1e7d71eorg.highwire.dtl.DTLVardef@1dc1252_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOFigure 1.C_FLOATNO Large mutation-associated datasets are used in a variety of experimental contexts. They can be used to map antibody footprints on viral glycoproteins, assess the impact of mutations on protein function in a laboratory setting, and identify patterns of selection from natural mutation frequencies.

C_FIG The mutation-based data generated by these approaches is often best understood in the context of a proteins 3D structure; for instance, to assess questions like how mutations that affect antibody escape relate to the physical antibody binding epitope on the protein. However, current approaches for visualizing mutation data in the context of a proteins structure are often cumbersome and require multiple steps and softwares. To streamline the visualization of mutation-associated data in the context of a protein structure, we developed a web-based tool, dms-viz. With dms-viz, users can straightforwardly visualize mutation-based data such as those from DMS experiments in the context of a 3D protein model in an interactive format. See https://dms-viz.github.io/ to use dms-viz.
]]></description>
<dc:creator>Hannon, W. W.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2023-11-01</dc:date>
<dc:identifier>doi:10.1101/2023.10.29.564578</dc:identifier>
<dc:title><![CDATA[dms-viz: Structure-informed visualizations for deep mutational scanning and other mutation-based datasets]]></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/10.1101/2023.10.30.564764v1?rss=1">
<title>
<![CDATA[
A statistical framework for powerful multi-trait rare variant analysis in large-scale whole-genome sequencing studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.30.564764v1?rss=1</link>
<description><![CDATA[
Large-scale whole-genome sequencing (WGS) studies have improved our understanding of the contributions of coding and noncoding rare variants to complex human traits. Leveraging association effect sizes across multiple traits in WGS rare variant association analysis can improve statistical power over single-trait analysis, and also detect pleiotropic genes and regions. Existing multi-trait methods have limited ability to perform rare variant analysis of large-scale WGS data. We propose MultiSTAAR, a statistical framework and computationally-scalable analytical pipeline for functionally-informed multi-trait rare variant analysis in large-scale WGS studies. MultiSTAAR accounts for relatedness, population structure and correlation among phenotypes by jointly analyzing multiple traits, and further empowers rare variant association analysis by incorporating multiple functional annotations. We applied MultiSTAAR to jointly analyze three lipid traits (low-density lipoprotein cholesterol, high-density lipoprotein cholesterol and triglycerides) in 61,861 multi-ethnic samples from the Trans-Omics for Precision Medicine (TOPMed) Program. We discovered new associations with lipid traits missed by single-trait analysis, including rare variants within an enhancer of NIPSNAP3A and an intergenic region on chromosome 1.
]]></description>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Selvaraj, M. S.</dc:creator>
<dc:creator>Van Buren, E.</dc:creator>
<dc:creator>Zhou, H.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Sun, R.</dc:creator>
<dc:creator>McCaw, Z. R.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Arnett, D. K.</dc:creator>
<dc:creator>Bis, J. C.</dc:creator>
<dc:creator>Blangero, J.</dc:creator>
<dc:creator>Boerwinkle, E.</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>Carson, A. P.</dc:creator>
<dc:creator>Carlson, J. C.</dc:creator>
<dc:creator>Chami, N.</dc:creator>
<dc:creator>Chen, Y.-D. I.</dc:creator>
<dc:creator>Curran, J. E.</dc:creator>
<dc:creator>de Vries, P. S.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Franceschini, N.</dc:creator>
<dc:creator>Freedman, B. I.</dc:creator>
<dc:creator>Gu, C.</dc:creator>
<dc:creator>Heard-Costa, N. L.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Hou, L.</dc:creator>
<dc:creator>Hung, Y.-J.</dc:creator>
<dc:creator>Irvin, M. R.</dc:creator>
<dc:creator>Kaplan, R. C.</dc:creator>
<dc:creator>Kardia, S. L. R.</dc:creator>
<dc:creator>Kelly, T.</dc:creator>
<dc:creator>Konigsberg, I.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Kral, B. G.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Loos, R. J. F.</dc:creator>
<dc:creator>Mahaney, M. C.</dc:creator>
<dc:creator>Martin, L. W.</dc:creator>
<dc:creator>Mathias, R. A.</dc:creator>
<dc:creator>Minster, R. L.</dc:creator>
<dc:creator>Mitchell, B. D</dc:creator>
<dc:date>2023-11-02</dc:date>
<dc:identifier>doi:10.1101/2023.10.30.564764</dc:identifier>
<dc:title><![CDATA[A statistical framework for powerful multi-trait rare variant analysis in large-scale whole-genome sequencing studies]]></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/10.1101/2023.11.01.565201v1?rss=1">
<title>
<![CDATA[
Small-molecule binding and sensing with a designed protein family 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.01.565201v1?rss=1</link>
<description><![CDATA[
Despite transformative advances in protein design with deep learning, the design of small-molecule-binding proteins and sensors for arbitrary ligands remains a grand challenge. Here we combine deep learning and physics-based methods to generate a family of proteins with diverse and designable pocket geometries, which we employ to computationally design binders for six chemically and structurally distinct small-molecule targets. Biophysical characterization of the designed binders revealed nanomolar to low micromolar binding affinities and atomic-level design accuracy. The bound ligands are exposed at one edge of the binding pocket, enabling the de novo design of chemically induced dimerization (CID) systems; we take advantage of this to create a biosensor with nanomolar sensitivity for cortisol. Our approach provides a general method to design proteins that bind and sense small molecules for a wide range of analytical, environmental, and biomedical applications.
]]></description>
<dc:creator>Lee, G. R.</dc:creator>
<dc:creator>Pellock, S. J.</dc:creator>
<dc:creator>Norn, C.</dc:creator>
<dc:creator>Tischer, D.</dc:creator>
<dc:creator>Dauparas, J.</dc:creator>
<dc:creator>Anishchenko, I.</dc:creator>
<dc:creator>Mercer, J. A. M.</dc:creator>
<dc:creator>Kang, A.</dc:creator>
<dc:creator>Bera, A.</dc:creator>
<dc:creator>Nguyen, H.</dc:creator>
<dc:creator>Goreshnik, I.</dc:creator>
<dc:creator>Vafeados, D.</dc:creator>
<dc:creator>Roullier, N.</dc:creator>
<dc:creator>Han, H. L.</dc:creator>
<dc:creator>Coventry, B.</dc:creator>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Liu, D. R.</dc:creator>
<dc:creator>Yeh, A. H.-W.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2023-11-02</dc:date>
<dc:identifier>doi:10.1101/2023.11.01.565201</dc:identifier>
<dc:title><![CDATA[Small-molecule binding and sensing with a designed protein family]]></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/10.1101/2023.11.05.565221v1?rss=1">
<title>
<![CDATA[
Evolutionary origin and structural ligand mimicry by the inserted domain of alpha-integrin proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.05.565221v1?rss=1</link>
<description><![CDATA[
Heterodimeric integrin proteins transmit signals through conformational changes upon ligand binding between their alpha () and beta ({beta}) subunits. Early in chordate evolution, some  subunits acquired an "inserted" (I) domain, which expanded their ligand binding capacity but simultaneously obstructed the ancestral ligand-binding pocket. While this would seemingly impede conventional ligand-mediated integrin activation, it was proposed that the I domain itself could serve both as a ligand replacement and an activation trigger. Here, we provide compelling evidence in support of this longstanding hypothesis using high-resolution cryo-electron microscopy structures of two distinct integrin complexes: the ligand-free and E-cadherin-bound states of the E{beta}7 integrin with the I domain, as well as the 4{beta}7 integrin lacking the I domain in both a ligand-free state and bound to MadCAM-1. We trace the evolutionary origin of the I domain to an ancestral collagen-collagen interaction domain. Our analyses illuminate how the I domain intrinsically mimics an extrinsic ligand, enabling integrins to undergo the canonical allosteric cascade of conformational activation and dramatically expanding the range of cellular communication mechanisms in vertebrates.
]]></description>
<dc:creator>Hollis, J. A.</dc:creator>
<dc:creator>Chan, M. C.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:creator>Campbell, M. G.</dc:creator>
<dc:date>2023-11-06</dc:date>
<dc:identifier>doi:10.1101/2023.11.05.565221</dc:identifier>
<dc:title><![CDATA[Evolutionary origin and structural ligand mimicry by the inserted domain of alpha-integrin proteins]]></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/10.1101/2023.11.13.566830v1?rss=1">
<title>
<![CDATA[
Infection pressure in apes has driven selection for CD4 alleles that resist lentivirus (HIV/SIV) infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.13.566830v1?rss=1</link>
<description><![CDATA[
Simian immunodeficiency viruses (SIVs) comprise a large group of primate lentiviruses that endemically infect African monkeys. HIV-1 spilled over to humans from this viral reservoir, but the spillover did not occur directly from monkeys to humans. Instead, a key event was the introduction of SIVs into great apes, which then set the stage for infection of humans. Here, we investigate the role of the lentiviral entry receptor, CD4, in this key and fateful event in the history of SIV/HIV emergence. First, we reconstructed and tested ancient forms of CD4 at two important nodes in ape speciation, both prior to the infection of chimpanzees and gorillas with these viruses. These ancestral CD4s fully supported entry of diverse SIV isolates related to the viruses that made this initial jump to apes. In stark contrast, modern chimpanzee and gorilla CD4 orthologs are more resistant to these viruses. To investigate how this resistance in CD4 was gained, we acquired CD4 gene sequences from 32 gorilla individuals of two species, and identified alleles that encode 8 unique CD4 protein variants. Functional testing of these identified variant-specific differences in susceptibility to virus entry. By engineering single point mutations from resistant gorilla CD4 variants into the permissive human CD4 receptor, we demonstrate that acquired substitutions in gorilla CD4 did convey resistance to virus entry. We provide a population genetic analysis to support the theory that selection is acting in favor of more and more resistant CD4 alleles in ape species harboring SIV endemically (gorillas and chimpanzees), but not in other ape species that lack SIV infections (bonobos and orangutans). Taken together, our results show that SIV has placed intense selective pressure on ape CD4, acting to propagate SIV-resistant alleles in chimpanzee and gorilla populations.
]]></description>
<dc:creator>Warren, C.</dc:creator>
<dc:creator>Barbachano-Guerrero, A.</dc:creator>
<dc:creator>DuMont, V.</dc:creator>
<dc:creator>Stabell, A.</dc:creator>
<dc:creator>Dirasantha, O.</dc:creator>
<dc:creator>Yang, Q.</dc:creator>
<dc:creator>Sawyer, S. L.</dc:creator>
<dc:date>2023-11-13</dc:date>
<dc:identifier>doi:10.1101/2023.11.13.566830</dc:identifier>
<dc:title><![CDATA[Infection pressure in apes has driven selection for CD4 alleles that resist lentivirus (HIV/SIV) infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.13.566961v1?rss=1">
<title>
<![CDATA[
Full-spike deep mutational scanning helps predict the evolutionary success of SARS-CoV-2 clades 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.13.566961v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 variants acquire mutations in spike that promote immune evasion and impact other properties that contribute to viral fitness such as ACE2 receptor binding and cell entry. Knowledge of how mutations affect these spike phenotypes can provide insight into the current and potential future evolution of the virus. Here we use pseudovirus deep mutational scanning to measure how >9,000 mutations across the full XBB.1.5 and BA.2 spikes affect ACE2 binding, cell entry, or escape from human sera. We find that mutations outside the receptor-binding domain (RBD) have meaningfully impacted ACE2 binding during SARS-CoV-2 evolution. We also measure how mutations to the XBB.1.5 spike affect neutralization by serum from individuals who recently had SARS-CoV-2 infections. The strongest serum escape mutations are in the RBD at sites 357, 420, 440, 456, and 473--however, the antigenic impacts of these mutations vary across individuals. We also identify strong escape mutations outside the RBD; however many of them decrease ACE2 binding, suggesting they act by modulating RBD conformation. Notably, the growth rates of human SARS-CoV-2 clades can be explained in substantial part by the measured effects of mutations on spike phenotypes, suggesting our data could enable better prediction of viral evolution.
]]></description>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Brown, J.</dc:creator>
<dc:creator>McMahon, T. E.</dc:creator>
<dc:creator>Farrell, A. G.</dc:creator>
<dc:creator>Asarnow, D.</dc:creator>
<dc:creator>Stewart, C.</dc:creator>
<dc:creator>Logue, J.</dc:creator>
<dc:creator>Murrell, B.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2023-11-14</dc:date>
<dc:identifier>doi:10.1101/2023.11.13.566961</dc:identifier>
<dc:title><![CDATA[Full-spike deep mutational scanning helps predict the evolutionary success of SARS-CoV-2 clades]]></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/10.1101/2023.11.11.566714v1?rss=1">
<title>
<![CDATA[
Expansion of human centromeric arrays in cells undergoing break-induced replication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.11.566714v1?rss=1</link>
<description><![CDATA[
Human centromeres are located within -satellite arrays and evolve rapidly, which can lead to individual variation in array lengths. Proposed mechanisms for such alterations in lengths are unequal cross-over between sister chromatids, gene conversion, and break-induced replication. However, the underlying molecular mechanisms responsible for the massive, complex, and homogeneous organization of centromeric arrays have not been experimentally validated. Here, we use droplet digital PCR assays to demonstrate that centromeric arrays can expand and contract within ~20 somatic cell divisions of a cell line. We find that the frequency of array variation among single-cell-derived subclones ranges from a minimum of ~7% to a maximum of ~100%. Further clonal evolution revealed that centromere expansion is favored over contraction. We find that the homologous recombination protein RAD52 and the helicase PIF1 are required for extensive array change, suggesting that centromere sequence evolution can occur via break-induced replication.
]]></description>
<dc:creator>Showman, S.</dc:creator>
<dc:creator>Talbert, P. B.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Adeyemi, R. O.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2023-11-15</dc:date>
<dc:identifier>doi:10.1101/2023.11.11.566714</dc:identifier>
<dc:title><![CDATA[Expansion of human centromeric arrays in cells undergoing break-induced replication]]></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/10.1101/2023.11.13.566946v1?rss=1">
<title>
<![CDATA[
High-throughput thermodynamic and kinetic measurements of transcription factor/DNA mutations reveal how conformational heterogeneity can shape motif selectivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.13.566946v1?rss=1</link>
<description><![CDATA[
Transcription factors (TFs) bind DNA sequences with a range of affinities, yet the mechanisms determining energetic differences between high- and low-affinity sequences ( selectivity) remain poorly understood. Here, we investigated two basic helix-loop-helix TFs, MAX (H. sapiens) and Pho4 (S. cerevisiae), that bind the same high-affinity sequence with highly similar nucleotide-contacting residues and bound structures but are differentially selective for non-cognate sequences. By measuring >1700 Kds and >500 rate constants for Pho4 and MAX mutant libraries binding multiple DNA sequences and comparing these measurements with thermodynamic and kinetic models, we identify the biophysical mechanisms by which changes to TF sequence alter both bound and unbound conformational ensembles to shape specificity landscapes. These results highlight the importance of conformational heterogeneity in determining sequence specificity and selectivity and can guide future efforts to engineer nucleic acid-binding proteins with enhanced selectivity.
]]></description>
<dc:creator>Fordyce, P. M.</dc:creator>
<dc:creator>Hastings, R.</dc:creator>
<dc:creator>Aditham, A.</dc:creator>
<dc:creator>DelRosso, N.</dc:creator>
<dc:creator>Suzuki, P.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.13.566946</dc:identifier>
<dc:title><![CDATA[High-throughput thermodynamic and kinetic measurements of transcription factor/DNA mutations reveal how conformational heterogeneity can shape motif selectivity]]></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/10.1101/2023.11.14.567084v1?rss=1">
<title>
<![CDATA[
Two Septin Complexes Mediate Actin Dynamics During Cell Wound Repair 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.14.567084v1?rss=1</link>
<description><![CDATA[
Cells have robust wound repair systems to prevent further damage or infection and to quickly restore cell cortex integrity when exposed to mechanical and chemical stress. Actomyosin ring formation and contraction at the wound edge are major events during closure of the plasma membrane and underlying cytoskeleton during cell wound repair. Here, we show that all five Drosophila Septins are required for efficient cell wound repair. Based on their different recruitment patterns and knockdown/mutant phenotypes, two distinct Septin complexes, Sep1-Sep2-Pnut and Sep4-Sep5-Pnut, are assembled to regulate actin ring assembly, contraction, and remodeling during the repair process. Intriguingly, we find that these two Septin complexes have different F-actin bending activities. In addition, we find that Anillin regulates the recruitment of only one of two Septin complexes upon wounding. Our results demonstrate that two functionally distinct Septin complexes work side-by-side to discretely regulate actomyosin ring dynamics during cell wound repair.
]]></description>
<dc:creator>Stjepic, V.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Hui, J.</dc:creator>
<dc:creator>Parkhurst, S. M.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.14.567084</dc:identifier>
<dc:title><![CDATA[Two Septin Complexes Mediate Actin Dynamics During Cell Wound Repair]]></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/10.1101/2023.11.16.567469v1?rss=1">
<title>
<![CDATA[
A novel non-invasive method to sample immune cells in the lower female genital tract 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.16.567469v1?rss=1</link>
<description><![CDATA[
T cells in the human female genital tract (FGT)2 are key mediators of susceptibility to and protection from infection, including HIV and other sexually transmitted infections. There is a critical need for increased understanding of the distribution and activation of T cell populations in the FGT, but current sampling methods require a healthcare provider and are expensive, limiting the ability to study these populations longitudinally. To address these challenges, we have developed a method to sample immune cells from the FGT utilizing disposable menstrual discs which are non-invasive, self-applied, and low-cost. To demonstrate reproducibility, we sampled the cervicovaginal fluid (CVF)3 of healthy, reproductive-aged individuals using menstrual discs over three sequential days. CVF was processed for cervicovaginal cells, and high parameter flow cytometry was used to characterize immune populations. We identified large numbers of live, CD45+ leukocytes, as well as distinct populations of T cells and B cells. Within the T cell compartment, activation and suppression status of T cell subsets were consistent with previous studies of the FGT utilizing current approaches, including identification of both tissue resident and migratory populations. In addition, the T cell population structure was highly conserved across days within individuals but divergent across individuals. Our approach to sample immune cells in the FGT with menstrual discs will decrease barriers to participation and empower longitudinal sampling in future research studies.
]]></description>
<dc:creator>Peters, M. Q.</dc:creator>
<dc:creator>Domenjo-Vila, E.</dc:creator>
<dc:creator>Carlson, M.</dc:creator>
<dc:creator>Edlefsen, P. T.</dc:creator>
<dc:creator>Gasper, M.</dc:creator>
<dc:creator>Dabee, S.</dc:creator>
<dc:creator>Armistead, B.</dc:creator>
<dc:creator>Whidbey, C.</dc:creator>
<dc:creator>Jaspan, H. B.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:creator>Harrington, W. E.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.16.567469</dc:identifier>
<dc:title><![CDATA[A novel non-invasive method to sample immune cells in the lower female genital tract]]></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/10.1101/2023.11.19.566520v1?rss=1">
<title>
<![CDATA[
Whole genome association testing in 333,100 individuals across three biobanks identifies rare non-coding single variant and genomic aggregate associations with height 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.19.566520v1?rss=1</link>
<description><![CDATA[
The role of rare non-coding variation in complex human phenotypes is still largely unknown. To elucidate the impact of rare variants in regulatory elements, we performed a whole-genome sequencing association analysis for height using 333,100 individuals from three datasets: UK Biobank (N=200,003), TOPMed (N=87,652) and All of Us (N=45,445). We performed rare (<0.1% minor-allele-frequency) single-variant and aggregate testing of non-coding variants in regulatory regions based on proximal, intergenic and deep-intronic annotation. We observed 29 independent variants associated with height at P < 6 x 10-10 after conditioning on previously reported variants, with effect sizes ranging from -7cm to +4.7cm. We also identified and replicated non-coding aggregate-based associations proximal to HMGA1 containing variants associated with a 5cm taller height and of highly-conserved variants in MIR497HG on chromosome 17. We have developed a novel approach for identifying non-coding rare variants in regulatory regions with large effects from whole-genome sequencing data associated with complex traits.
]]></description>
<dc:creator>Hawkes, G.</dc:creator>
<dc:creator>Beaumont, R. N.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Mandla, R.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Albert, C. M.</dc:creator>
<dc:creator>Arnett, D. K.</dc:creator>
<dc:creator>Ashley-Koch, A. E.</dc:creator>
<dc:creator>Ashrani, A. A.</dc:creator>
<dc:creator>Barnes, K. C.</dc:creator>
<dc:creator>Boerwinkle, E.</dc:creator>
<dc:creator>Brody, J. A.</dc:creator>
<dc:creator>Carson, A. P.</dc:creator>
<dc:creator>Chami, N.</dc:creator>
<dc:creator>Chen, Y.-D. I.</dc:creator>
<dc:creator>Chung, M. K.</dc:creator>
<dc:creator>Curran, J. E.</dc:creator>
<dc:creator>Darbar, D.</dc:creator>
<dc:creator>Ellinor, P. T.</dc:creator>
<dc:creator>Fornage, M.</dc:creator>
<dc:creator>Gordeuk, V. R.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Hwu, C.-M.</dc:creator>
<dc:creator>Kalyani, R. R.</dc:creator>
<dc:creator>Kaplan, R.</dc:creator>
<dc:creator>Kardia, S. L. R.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Loos, R. J. F.</dc:creator>
<dc:creator>Lubitz, S. A.</dc:creator>
<dc:creator>Minster, R. L.</dc:creator>
<dc:creator>Mitchell, B. D.</dc:creator>
<dc:creator>Murabito, J. M.</dc:creator>
<dc:creator>Palmer, N. D.</dc:creator>
<dc:creator>Psaty, B. M.</dc:creator>
<dc:creator>Redline, S.</dc:creator>
<dc:creator>Shoemaker, M. B.</dc:creator>
<dc:creator>Silverman, E. K.</dc:creator>
<dc:creator>Telen, M. J.</dc:creator>
<dc:creator>Weiss, S. T.</dc:creator>
<dc:creator>Yanek, L. R.</dc:creator>
<dc:creator>Zhou, H.</dc:creator>
<dc:creator>NH</dc:creator>
<dc:date>2023-11-20</dc:date>
<dc:identifier>doi:10.1101/2023.11.19.566520</dc:identifier>
<dc:title><![CDATA[Whole genome association testing in 333,100 individuals across three biobanks identifies rare non-coding single variant and genomic aggregate associations with height]]></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/10.1101/2023.11.30.569313v1?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/10.1101/2023.12.03.569799v1?rss=1">
<title>
<![CDATA[
Calcium influx rapidly establishes distinct spatial recruitments of Annexins to cell wounds 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.03.569799v1?rss=1</link>
<description><![CDATA[
To survive daily damage, the formation of actomyosin ring at the wound periphery is required to rapidly close cell wounds. Calcium influx is one of the start signals for these cell wound repair events. Here, we find that rapid recruitment of all three Drosophila calcium responding and phospholipid binding Annexin proteins (AnxB9, AnxB10, AnxB11) to distinct regions around the wound are regulated by the quantity of calcium influx rather than their binding to specific phospholipids. The distinct recruitment patterns of these Annexins regulate the subsequent recruitment of RhoGEF2 and RhoGEF3 through actin stabilization to form a robust actomyosin ring. Surprisingly, we find that reduced extracellular calcium and depletion of intracellular calcium affect cell wound repair differently, despite these two conditions exhibiting similar GCaMP signals. Thus, our results suggest that, in addition to initiating repair events, both the quantity and sources of calcium influx are important for precise Annexin spatiotemporal protein recruitment to cell wounds and efficient wound repair.

SummaryCells have rapid and robust repair systems to survive daily damage. This study shows that calcium influx regulates the three distinct Drosophila Annexin recruitment patterns to the cell wound in order to organize an actomyosin ring for efficient wound closure.
]]></description>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Parkhurst, S. M.</dc:creator>
<dc:date>2023-12-04</dc:date>
<dc:identifier>doi:10.1101/2023.12.03.569799</dc:identifier>
<dc:title><![CDATA[Calcium influx rapidly establishes distinct spatial recruitments of Annexins to cell wounds]]></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/10.1101/2023.12.07.570700v1?rss=1">
<title>
<![CDATA[
snPATHO-seq: unlocking the pathology archives 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.07.570700v1?rss=1</link>
<description><![CDATA[
Formalin-fixed paraffin-embedded (FFPE) samples are valuable but underutilized in single-cell omics research due to their low DNA and RNA quality. In this study, leveraging recent single-cell genomic technology advances, we introduce a versatile method to derive high-quality single-nucleus transcriptomic data from FFPE samples.
]]></description>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Harvey, K.</dc:creator>
<dc:creator>Escudero Morales, J.</dc:creator>
<dc:creator>Kiedik, B.</dc:creator>
<dc:creator>Al-Eryani, G.</dc:creator>
<dc:creator>Greenwald, A. C.</dc:creator>
<dc:creator>Kalavros, N.</dc:creator>
<dc:creator>SegatovDezem, F.</dc:creator>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Pita-Juarez, Y.</dc:creator>
<dc:creator>Wise, K.</dc:creator>
<dc:creator>Degletagne, C.</dc:creator>
<dc:creator>Elz, A.</dc:creator>
<dc:creator>Johanneson, J.</dc:creator>
<dc:creator>Pakiam, F.</dc:creator>
<dc:creator>Ryu, H.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Tonon, L.</dc:creator>
<dc:creator>Kohlway, A.</dc:creator>
<dc:creator>Drennon, T.</dc:creator>
<dc:creator>Hadadianpour, A.</dc:creator>
<dc:creator>Abousoud, J.</dc:creator>
<dc:creator>Scott, R.</dc:creator>
<dc:creator>Lund, P.</dc:creator>
<dc:creator>Durruthy, J.</dc:creator>
<dc:creator>Vallejos, A. F.</dc:creator>
<dc:creator>Kaczorowski, D.</dc:creator>
<dc:creator>Warren, J.</dc:creator>
<dc:creator>Butler, L. M.</dc:creator>
<dc:creator>O'Toole, S.</dc:creator>
<dc:creator>Plummer, J. T.</dc:creator>
<dc:creator>Vlachos, I.</dc:creator>
<dc:creator>Lundeberg, J.</dc:creator>
<dc:creator>Swarbrick, A.</dc:creator>
<dc:creator>Martelotto, L. G.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.07.570700</dc:identifier>
<dc:title><![CDATA[snPATHO-seq: unlocking the pathology archives]]></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/10.1101/2023.12.07.570711v1?rss=1">
<title>
<![CDATA[
Viral gene drive spread during herpes simplex virus 1 infection in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.07.570711v1?rss=1</link>
<description><![CDATA[
Gene drives are genetic modifications designed to propagate efficiently through a population. Most applications rely on homologous recombination during sexual reproduction in diploid organisms such as insects, but we recently developed a gene drive in herpesviruses that relies on co-infection of cells by wild-type and engineered viruses. Here, we developed a viral gene drive against human herpes simplex virus 1 (HSV-1) and showed that it propagated efficiently in vitro and during HSV-1 infection in mice. We observed high levels of co-infection and gene drive-mediated recombination in neuronal tissues during herpes encephalitis as the infection progressed from the site of inoculation to the peripheral and central nervous systems. In addition, we found evidence that a superinfecting gene drive virus could recombine with wild-type viruses during latent infection. These findings indicated that HSV-1 achieves high rates of co-infection and recombination during viral infection, a phenomenon that is currently underappreciated. Overall, this study showed that a viral gene drive could spread in vivo during HSV-1 infection, paving the way toward therapeutic applications.
]]></description>
<dc:creator>Walter, M.</dc:creator>
<dc:creator>Riley, R.</dc:creator>
<dc:creator>Haick, A. K.</dc:creator>
<dc:creator>Massa, P. A.</dc:creator>
<dc:creator>Strongin, D. E.</dc:creator>
<dc:creator>Klouser, L. M.</dc:creator>
<dc:creator>Loprieno, M. A.</dc:creator>
<dc:creator>Stensland, L.</dc:creator>
<dc:creator>Santo, T. K.</dc:creator>
<dc:creator>Taylor, M. P.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Verdin, E.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.07.570711</dc:identifier>
<dc:title><![CDATA[Viral gene drive spread during herpes simplex virus 1 infection in mice]]></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/10.1101/2023.12.09.570874v1?rss=1">
<title>
<![CDATA[
SIRPα controls CD47-dependent platelet clearance in mice and humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.09.570874v1?rss=1</link>
<description><![CDATA[
Over the last decade, more data has revealed that increased surface expression of the "dont eat me" CD47 protein on cancer cells plays a role in immune evasion and tumor progression, with CD47 blockade emerging as a new therapy in immuno-oncology. CD47 is critical in regulating cell homeostasis and clearance, as binding of CD47 to the inhibitory receptor SIRP can prevent phagocytosis and macrophage-mediated cell clearance. The purpose of this study was to examine the role of the CD47-SIRP signal in platelet homeostasis and clearance. Therapeutic reagents targeting the CD47-SIRP axis are very promising for treatment of hematologic malignancies and solid tumors, but lead to transient anemia or thrombocytopenia in a subset of patients. We found that platelet homeostatic clearance is regulated through the CD47-SIRP axis and that therapeutic blockade to disrupt this interaction in mice and in humans has a significant impact on platelet levels. Furthermore, we identified genetic variations at the SIRPA locus that impact platelet levels in humans such that higher SIRPA gene expression is associated with higher platelet levels. SIRPA expression at either end of the normal range may affect clinical outcomes of treatment with anti-CD47 therapy.

Key pointsO_LIPlatelet homeostasis is regulated through the CD47-SIRP axis and therapeutic blockade to disrupt this interaction impacts platelet levels
C_LIO_LICommon genetic variants at SIRPA locus associate with platelet levels
C_LI
]]></description>
<dc:creator>Shoham, M.</dc:creator>
<dc:creator>Yiu, Y. Y.</dc:creator>
<dc:creator>Hansen, P.</dc:creator>
<dc:creator>Subramaniam, A.</dc:creator>
<dc:creator>Broberg, M.</dc:creator>
<dc:creator>Gars, E.</dc:creator>
<dc:creator>Raveh, T.</dc:creator>
<dc:creator>FinnGen,</dc:creator>
<dc:creator>Weissman, I. L.</dc:creator>
<dc:creator>Sinnott-Armstrong, N.</dc:creator>
<dc:creator>Krishnan, A.</dc:creator>
<dc:creator>Ollila, H.</dc:creator>
<dc:creator>Tal, M. C.</dc:creator>
<dc:date>2023-12-10</dc:date>
<dc:identifier>doi:10.1101/2023.12.09.570874</dc:identifier>
<dc:title><![CDATA[SIRPα controls CD47-dependent platelet clearance in mice and humans]]></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/10.1101/2023.12.14.571745v1?rss=1">
<title>
<![CDATA[
50-color phenotyping of the human immune system with in-depth assessment of T cells and dendritic cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.14.571745v1?rss=1</link>
<description><![CDATA[
We report the development of an optimized 50-color spectral flow cytometry panel designed for the in-depth analysis of the immune system in human blood and tissues, with the goal of maximizing the amount of information that can be collected using currently available flow cytometry platforms. We established and tested this panel using peripheral blood mononuclear cells (PBMCs), but included CD45 to enable its use for the analysis of human tissue samples. The panel contains lineage markers for all major immune cell subsets, and an extensive set of phenotyping markers focused on the activation and differentiation status of the T cell and dendritic cell (DC) compartment.

We outline the biological insight that can be gained from the simultaneous measurement of such a large number of proteins and propose that this approach provides a unique opportunity for the comprehensive exploration of the immune status in tissue biopsies and other human samples with a limited number of cells. Of note, we tested the panel to be compatible with cell sorting for further downstream applications. Furthermore, to facilitate the wide-spread implementation of such a panel across different cohorts and samples, we established a trimmed-down 45-color version which can be used with different spectral cytometry platforms.

Finally, to generate this panel, we utilized not only existing panel design guidelines, but also developed new metrics to systematically identify the optimal combination of 50 fluorochromes and evaluate fluorochrome-specific resolution in the context of a 50-color unmixing matrix.
]]></description>
<dc:creator>Konecny, A. J.</dc:creator>
<dc:creator>Mage, P.</dc:creator>
<dc:creator>Tyznik, A. J.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:creator>Mair, F.</dc:creator>
<dc:date>2023-12-15</dc:date>
<dc:identifier>doi:10.1101/2023.12.14.571745</dc:identifier>
<dc:title><![CDATA[50-color phenotyping of the human immune system with in-depth assessment of T cells and dendritic cells]]></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/10.1101/2023.12.29.573681v1?rss=1">
<title>
<![CDATA[
KMT2A oncoproteins induce epigenetic resistance to targeted therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.29.573681v1?rss=1</link>
<description><![CDATA[
Chromosomal translocations involving the Lysine-Methyl-Transferase-2A (KMT2A) locus generate potent oncogenic fusion proteins (oncoproteins) that disrupt regulation of developmental gene expression. By profiling the oncoprotein-target sites of 36 broadly representative KMT2A-rearranged leukemia samples, including three samples that underwent a lymphoid-to-myeloid lineage-switching event in response to therapy, we find the genomic enrichment of the oncoprotein is highly variable between samples and subject to dynamic regulation. At high levels of expression, the oncoproteins preferentially activate either an acute lymphoblastic leukemia (ALL) program, enriched for pro-B-cell genes, or an acute myeloid leukemia (AML) program, enriched for hematopoietic-stem-cell genes. The fusion-partner-specific-binding patterns over these gene sets are highly correlated with the prevalence of each mutation in ALL versus AML. In lineage-switching samples the oncoprotein levels are reduced and the oncoproteins preferentially activate granulocyte-monocyte progenitor (GMP) genes. In a sample that lineage switched during treatment with the menin inhibitor revumenib, the oncoprotein and menin are reduced to undetectable levels, but ENL, a transcriptional cofactor of the oncoprotein, persists on numerous oncoprotein-target loci, including genes in the GMP-like lineage-switching program. We propose KMT2A oncoproteins promote lineage-switching events through dynamic chromatin binding and can induce epigenetic lesions, marked by ENL, that support resistance to targeted therapies.
]]></description>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Duran, M.</dc:creator>
<dc:creator>Otto, D. J.</dc:creator>
<dc:creator>Kirkey, D.</dc:creator>
<dc:creator>Yi, J. S.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Sarthy, J. F.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2023-12-30</dc:date>
<dc:identifier>doi:10.1101/2023.12.29.573681</dc:identifier>
<dc:title><![CDATA[KMT2A oncoproteins induce epigenetic resistance to targeted therapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.17.576106v1?rss=1">
<title>
<![CDATA[
LSD1 inhibition suppresses ASCL1 and de-represses YAP1 to drive potent activity against neuroendocrine prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.17.576106v1?rss=1</link>
<description><![CDATA[
Lysine-specific demethylase 1 (LSD1 or KDM1A) has emerged as a critical mediator of tumor progression in metastatic castration-resistant prostate cancer (mCRPC). Among mCRPC subtypes, neuroendocrine prostate cancer (NEPC) is an exceptionally aggressive variant driven by lineage plasticity, an adaptive resistance mechanism to androgen receptor axis-targeted therapies. Our study shows that LSD1 expression is elevated in NEPC and associated with unfavorable clinical outcomes. Using genetic approaches, we validated the on-target effects of LSD1 inhibition across various models. We investigated the therapeutic potential of bomedemstat, an orally bioavailable, irreversible LSD1 inhibitor with low nanomolar potency. Our findings demonstrate potent antitumor activity against CRPC models, including tumor regressions in NEPC patient-derived xenografts. Mechanistically, our study uncovers that LSD1 inhibition suppresses the neuronal transcriptional program by downregulating ASCL1 through disrupting LSD1:INSM1 interactions and de-repressing YAP1 silencing. Our data support the clinical development of LSD1 inhibitors for treating CRPC - especially the aggressive NE phenotype.

Statement of SignificanceNeuroendocrine prostate cancer presents a clinical challenge due to the lack of effective treatments. Our research demonstrates that bomedemstat, a potent and selective LSD1 inhibitor, effectively combats neuroendocrine prostate cancer by downregulating the ASCL1- dependent NE transcriptional program and re-expressing YAP1.
]]></description>
<dc:creator>Mandl, A.</dc:creator>
<dc:creator>Jasmine, S.</dc:creator>
<dc:creator>Krueger, T. E. G.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Coleman, I. M.</dc:creator>
<dc:creator>Dalrymple, S. L.</dc:creator>
<dc:creator>Antony, L.</dc:creator>
<dc:creator>Rosen, D. M.</dc:creator>
<dc:creator>Jing, Y.</dc:creator>
<dc:creator>Hanratty, B.</dc:creator>
<dc:creator>Patel, R. A.</dc:creator>
<dc:creator>Low, J.-Y.</dc:creator>
<dc:creator>Dias, J.</dc:creator>
<dc:creator>Celatka, C. A.</dc:creator>
<dc:creator>Tapper, A. E.</dc:creator>
<dc:creator>Kleppe, M.</dc:creator>
<dc:creator>Kanayama, M.</dc:creator>
<dc:creator>Speranzini, V.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Corey, E.</dc:creator>
<dc:creator>Sena, L. A.</dc:creator>
<dc:creator>Casero, R. A.</dc:creator>
<dc:creator>Lotan, T.</dc:creator>
<dc:creator>Trock, B. J.</dc:creator>
<dc:creator>Kachhap, S. K.</dc:creator>
<dc:creator>Denmeade, S. R.</dc:creator>
<dc:creator>Carducci, M. A.</dc:creator>
<dc:creator>Mattevi, A.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Rienhoff, H. H.</dc:creator>
<dc:creator>Isaacs, J. T.</dc:creator>
<dc:creator>Brennen, W. N.</dc:creator>
<dc:date>2024-01-22</dc:date>
<dc:identifier>doi:10.1101/2024.01.17.576106</dc:identifier>
<dc:title><![CDATA[LSD1 inhibition suppresses ASCL1 and de-represses YAP1 to drive potent activity against neuroendocrine prostate cancer]]></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/10.1101/2024.02.21.581471v1?rss=1">
<title>
<![CDATA[
CYpHER: Catalytic extracellular targeted protein degradation with high potency and durable effect 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.21.581471v1?rss=1</link>
<description><![CDATA[
Many disease-causing proteins have multiple pathogenic mechanisms, and conventional inhibitors struggle to reliably disrupt more than one. Targeted protein degradation (TPD) can eliminate the protein, and thus all its functions, by directing a cells protein turnover machinery towards it. Two established strategies either engage catalytic E3 ligases or drive uptake towards the endolysosomal pathway. Here we describe CYpHER (CatalYtic pH-dependent Endolysosomal delivery with Recycling) technology with potency and durability from a novel catalytic mechanism that shares the specificity and straightforward modular design of endolysosomal uptake. By bestowing pH-dependent release on the target engager and using the rapid-cycling transferrin receptor as the uptake receptor, CYpHER induces endolysosomal target delivery while re-using drug, potentially yielding increased potency and reduced off-target tissue exposure risks. The TfR-based approach allows targeting to tumors that overexpress this receptor and offers the potential for transport to the CNS. CYpHER function was demonstrated in vitro with EGFR and PD-L1, and in vivo with EGFR in a model of EGFR-driven non-small cell lung cancer.
]]></description>
<dc:creator>Crook, Z. R.</dc:creator>
<dc:creator>Sevilla, G. P.</dc:creator>
<dc:creator>Young, P.</dc:creator>
<dc:creator>Girard, E. J.</dc:creator>
<dc:creator>Phi, T.-D.</dc:creator>
<dc:creator>Howard, M.</dc:creator>
<dc:creator>Olson, J. M.</dc:creator>
<dc:creator>Nairn, N. W.</dc:creator>
<dc:date>2024-02-23</dc:date>
<dc:identifier>doi:10.1101/2024.02.21.581471</dc:identifier>
<dc:title><![CDATA[CYpHER: Catalytic extracellular targeted protein degradation with high potency and durable effect]]></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/10.1101/2024.03.03.583199v1?rss=1">
<title>
<![CDATA[
Prevalent fast evolution of genes involved in heterochromatin functions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.03.583199v1?rss=1</link>
<description><![CDATA[
Heterochromatin is a gene-poor and repeat-rich genomic compartment universally found in eukaryotes. Despite its low transcriptional activity, heterochromatin plays important roles in maintaining genome stability, organizing chromosomes, and suppressing transposable elements (TEs). Given the importance of these functions, it is expected that the genes involved in heterochromatin regulation would be highly conserved. Yet, a handful of these genes were found to evolve rapidly. To investigate whether these previous findings are anecdotal or general to genes modulating heterochromatin, we compile an exhaustive list of 106 candidate genes involved in heterochromatin functions and investigate their evolution over short and long evolutionary time scales in Drosophila. Our analyses find that these genes exhibit significantly more frequent evolutionary changes, both in the forms of amino acid substitutions and gene copy number change, when compared to genes involved in Polycomb-based repressive chromatin. While positive selection drives amino acid changes within both structured domains with diverse functions and intrinsically disordered regions (IDRs), purifying selection may have maintained the proportions of IDRs of these proteins. Together with the observed negative associations between evolutionary rates of these genes and genomic TE abundance, we propose an evolutionary model where the fast evolution of genes involved in heterochromatin functions is an inevitable outcome of the unique functional roles of heterochromatin, while the rapid evolution of TEs may be an effect rather than cause. Our study provides an important global view of the evolution of genes involved in this critical cellular domain and provides insights into the factors driving the distinctive evolution of heterochromatin.
]]></description>
<dc:creator>Lin, L.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>McIntyre, J.</dc:creator>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Colmenares, S. S.</dc:creator>
<dc:creator>Lee, G. Y. C.</dc:creator>
<dc:date>2024-03-06</dc:date>
<dc:identifier>doi:10.1101/2024.03.03.583199</dc:identifier>
<dc:title><![CDATA[Prevalent fast evolution of genes involved in heterochromatin functions]]></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/10.1101/2024.03.08.584117v1?rss=1">
<title>
<![CDATA[
A Spatial Omnibus Test (SPOT) for Spatial Proteomic Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.08.584117v1?rss=1</link>
<description><![CDATA[
Spatial proteomics can reveal the spatial organization of immune cells in the tumor immune microenvironment. Relating measures of spatial clustering, such as Ripleys K or Besags L, to patient outcomes may offer important clinical insights. However, these measures require pre-specifying a radius in which to quantify clustering, yet no consensus exists on the optimal radius which may be context-specific. We propose a SPatial Omnibus Test (SPOT) which conducts this analysis across a range of candidate radii. At each radius, SPOT evaluates the association between the spatial summary and outcome, adjusting for confounders. SPOT then aggregates results across radii using the Cauchy combination test, yielding an omnibus p-value characterizing the overall degree of association. Using simulations, we verify that the type I error rate is controlled and show SPOT can be more powerful than alternatives. We also apply SPOT to an ovarian cancer study. An R package and tutorial is provided at https://github.com/sarahsamorodnitsky/SPOT.
]]></description>
<dc:creator>Samorodnitsky, S. N.</dc:creator>
<dc:creator>Campbell, K. M.</dc:creator>
<dc:creator>Ribas, A.</dc:creator>
<dc:creator>Wu, M. C.</dc:creator>
<dc:date>2024-03-13</dc:date>
<dc:identifier>doi:10.1101/2024.03.08.584117</dc:identifier>
<dc:title><![CDATA[A Spatial Omnibus Test (SPOT) for Spatial Proteomic Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.08.584120v1?rss=1">
<title>
<![CDATA[
Variant mutation in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.08.584120v1?rss=1</link>
<description><![CDATA[
The evolution of SARS-CoV-2 variants and their respective phenotypes represents an important set of tools to understand basic coronavirus biology as well as the public health implications of individual mutations in variants of concern. While mutations outside of Spike are not well studied, the entire viral genome is undergoing evolutionary selection, particularly the central disordered linker region of the nucleocapsid (N) protein. Here, we identify a mutation (G215C), characteristic of the Delta variant, that introduces a novel cysteine into this linker domain, which results in the formation of a disulfide bond and a stable N-N dimer. Using reverse genetics, we determined that this cysteine residue is necessary and sufficient for stable dimer formation in a WA1 SARS-CoV-2 background, where it results in significantly increased viral growth both in vitro and in vivo. Finally, we demonstrate that the N:G215C virus packages more nucleocapsid per virion and that individual virions are larger, with elongated morphologies.
]]></description>
<dc:creator>Kubinski, H. C.</dc:creator>
<dc:creator>Despres, H. W.</dc:creator>
<dc:creator>Johnson, B. A.</dc:creator>
<dc:creator>Schmidt, M. M.</dc:creator>
<dc:creator>Jaffrani, S. A.</dc:creator>
<dc:creator>Mills, M. G.</dc:creator>
<dc:creator>Lokugamage, K.</dc:creator>
<dc:creator>Dumas, C. M.</dc:creator>
<dc:creator>Shirley, D. J.</dc:creator>
<dc:creator>Estes, L. K.</dc:creator>
<dc:creator>Pekosz, A.</dc:creator>
<dc:creator>Crothers, J. W.</dc:creator>
<dc:creator>Roychoudhury, P.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Martorelli Di Genova, B.</dc:creator>
<dc:creator>Walker, D. H.</dc:creator>
<dc:creator>Ballif, B. A.</dc:creator>
<dc:creator>Ladinsky, M. S.</dc:creator>
<dc:creator>Bjorkman, P. J.</dc:creator>
<dc:creator>Menachery, V. D.</dc:creator>
<dc:creator>Bruce, E. A.</dc:creator>
<dc:date>2024-03-11</dc:date>
<dc:identifier>doi:10.1101/2024.03.08.584120</dc:identifier>
<dc:title><![CDATA[Variant mutation in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation]]></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/10.1101/2024.03.08.584165v1?rss=1">
<title>
<![CDATA[
Transcriptional bursting, gene activation, and roles of SAGA and Mediator Tail measured using nucleotide recoding single cell RNA-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.08.584165v1?rss=1</link>
<description><![CDATA[
A time resolved nascent single-cell RNA-seq approach was developed to dissect gene-specific transcriptional bursting and the roles of SAGA and Mediator Tail (the activator-binding module). Most yeast genes show near-constitutive behavior while only a subset of genes show high mRNA variance suggestive of transcription bursting. Bursting behavior is highest in the coactivator redundant (CR) gene class (dependent on both SAGA and TFIID) and is strongest in TATA-containing CR genes. Applying this approach to analyze gene activation, we found that basal histone gene transcription is in a low level, low-noise constitutive mode while the activated state unexpectedly shows an increase in both the fraction of active promoters and a switch to a noisy and bursty transcription mode. Rapid depletion of either SAGA or Mediator Tail suggests that both factors play an important role in stimulating the fraction of active promoters at CR genes, with a variable gene-specific role in transcriptional bursting.
]]></description>
<dc:creator>Schofield, J. A.</dc:creator>
<dc:creator>Hahn, S.</dc:creator>
<dc:date>2024-03-13</dc:date>
<dc:identifier>doi:10.1101/2024.03.08.584165</dc:identifier>
<dc:title><![CDATA[Transcriptional bursting, gene activation, and roles of SAGA and Mediator Tail measured using nucleotide recoding single cell RNA-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.08.584176v1?rss=1">
<title>
<![CDATA[
High-throughput sequencing-based neutralization assay reveals how repeated vaccinations impact titers to recent human H1N1 influenza strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.08.584176v1?rss=1</link>
<description><![CDATA[
The high genetic diversity of influenza viruses means that traditional serological assays have too low throughput to measure serum antibody neutralization titers against all relevant strains. To overcome this challenge, we have developed a sequencing-based neutralization assay that simultaneously measures titers against many viral strains using small serum volumes via a workflow similar to traditional neutralization assays. The key innovation is to incorporate unique nucleotide barcodes into the hemagglutinin (HA) genomic segment, and then pool viruses with numerous different barcoded HA variants and quantify infectivity of all of them simultaneously using next-generation sequencing. With this approach, a single researcher performed the equivalent of 2,880 traditional neutralization assays (80 serum samples against 36 viral strains) in approximately one month. We applied the sequencing-based assay to quantify the impact of influenza vaccination on neutralization titers against recent human H1N1 strains for individuals who had or had not also received a vaccine in the previous year. We found that the viral strain specificities of the neutralizing antibodies elicited by vaccination vary among individuals, and that vaccination induced a smaller increase in titers for individuals who had also received a vaccine the previous year--although the titers six months after vaccination were similar in individuals with and without the previous-year vaccination. We also identified a subset of individuals with low titers to a subclade of recent H1N1 even after vaccination. This study demonstrates the utility of high-throughput sequencing-based neutralization assays that enable titers to be simultaneously measured against many different viral strains. We provide a detailed experimental protocol (DOI: https://dx.doi.org/10.17504/protocols.io.kqdg3xdmpg25/v1) and a computational pipeline (https://github.com/jbloomlab/seqneut-pipeline) for the sequencing-based neutralization assays to facilitate the use of this method by others.
]]></description>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Tarabi, R. A. L.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Touyon, L.</dc:creator>
<dc:creator>Wong, S. S.</dc:creator>
<dc:creator>Cheng, S. M.</dc:creator>
<dc:creator>Leung, N. H.</dc:creator>
<dc:creator>Hannon, W. W.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Cobey, S.</dc:creator>
<dc:creator>Cowling, B. J.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2024-03-09</dc:date>
<dc:identifier>doi:10.1101/2024.03.08.584176</dc:identifier>
<dc:title><![CDATA[High-throughput sequencing-based neutralization assay reveals how repeated vaccinations impact titers to recent human H1N1 influenza strains]]></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/10.1101/2024.03.13.584735v1?rss=1">
<title>
<![CDATA[
Protein nanoparticle vaccines induce potent neutralizing antibody responses against MERS-CoV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.13.584735v1?rss=1</link>
<description><![CDATA[
Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic betacoronavirus that causes severe and often lethal respiratory illness in humans. The MERS-CoV spike (S) protein is the viral fusogen and the target of neutralizing antibodies, and has therefore been the focus of vaccine design efforts. Currently there are no licensed vaccines against MERS-CoV and only a few candidates have advanced to Phase I clinical trials. Here we developed MERS-CoV vaccines utilizing a computationally designed protein nanoparticle platform that has generated safe and immunogenic vaccines against various enveloped viruses, including a licensed vaccine for SARS-CoV-2. Two-component protein nanoparticles displaying MERS-CoV S-derived antigens induced robust neutralizing antibody responses and protected mice against challenge with mouse-adapted MERS-CoV. Electron microscopy polyclonal epitope mapping and serum competition assays revealed the specificities of the dominant antibody responses elicited by immunogens displaying the prefusion-stabilized S-2P trimer, receptor binding domain (RBD), or N-terminal domain (NTD). An RBD nanoparticle vaccine elicited antibodies targeting multiple non-overlapping epitopes in the RBD, whereas anti-NTD antibodies elicited by the S-2P- and NTD-based immunogens converged on a single antigenic site. Our findings demonstrate the potential of two-component nanoparticle vaccine candidates for MERS-CoV and suggest that this platform technology could be broadly applicable to betacoronavirus vaccine development.
]]></description>
<dc:creator>Chao, C. W.</dc:creator>
<dc:creator>Sprouse, K. R.</dc:creator>
<dc:creator>Miranda, M. C.</dc:creator>
<dc:creator>Catanzaro, N. J.</dc:creator>
<dc:creator>Hubbard, M. L.</dc:creator>
<dc:creator>Addetia, A.</dc:creator>
<dc:creator>Stewart, C.</dc:creator>
<dc:creator>Brown, J. T.</dc:creator>
<dc:creator>Dosey, A.</dc:creator>
<dc:creator>Valdez, A.</dc:creator>
<dc:creator>Ravichandran, R.</dc:creator>
<dc:creator>Hendricks, G. G.</dc:creator>
<dc:creator>Ahlrichs, M.</dc:creator>
<dc:creator>Dobbins, C.</dc:creator>
<dc:creator>Hand, A.</dc:creator>
<dc:creator>Treichel, C.</dc:creator>
<dc:creator>Willoughby, I.</dc:creator>
<dc:creator>Walls, A. C.</dc:creator>
<dc:creator>McGuire, A. T.</dc:creator>
<dc:creator>Leaf, E. M.</dc:creator>
<dc:creator>Baric, R. S.</dc:creator>
<dc:creator>Schaefer, A.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:date>2024-03-14</dc:date>
<dc:identifier>doi:10.1101/2024.03.13.584735</dc:identifier>
<dc:title><![CDATA[Protein nanoparticle vaccines induce potent neutralizing antibody responses against MERS-CoV]]></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/10.1101/2024.03.13.584849v1?rss=1">
<title>
<![CDATA[
Development of a series of genetically engineered NTRK fusion-driven pediatric-type high-grade glioma mouse models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.13.584849v1?rss=1</link>
<description><![CDATA[
Pediatric-type high-grade gliomas frequently harbor gene fusions involving receptor tyrosine kinase genes, including neurotrophic tyrosine kinase receptor (NTRK) fusions. Clinically, these tumors show high initial response rates to tyrosine kinase inhibition but ultimately recur due to the accumulation of additional resistance-conferring mutations. Here, we developed a series of genetically engineered mouse models of treatment-naive and -experienced NTRK1/2/3 fusion-driven gliomas. Both the TRK kinase domain and the N-terminal fusion partners influenced tumor histology and aggressiveness. Treatment with TRK kinase inhibitors significantly extended survival of NTRK fusion-driven glioma mice in a fusion- and inhibitor-dependent manner, but tumors ultimately recurred due to the presence of treatment-resistant persister cells. Finally, we show that ERK activation promotes resistance to TRK kinase inhibition and identify MEK inhibition as a potential combination therapy. These models will be invaluable tools for preclinical testing of novel inhibitors and to study the cellular responses of NTRK fusion-driven gliomas to therapy.
]]></description>
<dc:creator>Schmid, S.</dc:creator>
<dc:creator>Russell, Z. R.</dc:creator>
<dc:creator>Yamashita, A. S.</dc:creator>
<dc:creator>West, M. E.</dc:creator>
<dc:creator>Parrish, A. G.</dc:creator>
<dc:creator>Walker, J.</dc:creator>
<dc:creator>Rudoy, D.</dc:creator>
<dc:creator>Yan, J. Z.</dc:creator>
<dc:creator>Quist, D. C.</dc:creator>
<dc:creator>Gessesse, B. N.</dc:creator>
<dc:creator>Alvinez, N.</dc:creator>
<dc:creator>Cimino, P. J.</dc:creator>
<dc:creator>Kumasaka, D. K.</dc:creator>
<dc:creator>Parchment, R. E.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:date>2024-03-13</dc:date>
<dc:identifier>doi:10.1101/2024.03.13.584849</dc:identifier>
<dc:title><![CDATA[Development of a series of genetically engineered NTRK fusion-driven pediatric-type high-grade glioma mouse models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.21.586113v1?rss=1">
<title>
<![CDATA[
Sir2 and Fun30 regulate ribosomal DNA replication timing via Mcm helicase positioning and nucleosome occupancy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.21.586113v1?rss=1</link>
<description><![CDATA[
The association between late replication timing and low transcription rates in eukaryotic heterochromatin is well-known, yet the specific mechanisms underlying this link remain uncertain. In Saccharomyces cerevisiae, the histone deacetylase Sir2 is required for both transcriptional silencing and late replication at the repetitive ribosomal DNA arrays (rDNA). We have previously reported that in the absence of SIR2, a derepressed RNA PolII repositions MCM replicative helicases from their loading site at the ribosomal origin, where they abut well-positioned, high-occupancy nucleosomes, to an adjacent region with lower nucleosome occupancy. By developing a method that can distinguish activation of closely spaced MCM complexes, here we show that the displaced MCMs at rDNA origins have increased firing propensity compared to the nondisplaced MCMs. Furthermore, we found that both, activation of the repositioned MCMs and low occupancy of the adjacent nucleosomes critically depend on the chromatin remodeling activity of FUN30. Our study elucidates the mechanism by which Sir2 delays replication timing, and it demonstrates, for the first time, that activation of a specific replication origin in vivo relies on the nucleosome context shaped by a single chromatin remodeler.
]]></description>
<dc:creator>Lichauco, C.</dc:creator>
<dc:creator>Foss, E. J.</dc:creator>
<dc:creator>Gatbonton-Schwager, T.</dc:creator>
<dc:creator>Athow, N. F.</dc:creator>
<dc:creator>Lofts, B.</dc:creator>
<dc:creator>Acob, R.</dc:creator>
<dc:creator>Taylor, E.</dc:creator>
<dc:creator>Lao, U.</dc:creator>
<dc:creator>Miles, s.</dc:creator>
<dc:creator>Bedalov, A.</dc:creator>
<dc:date>2024-03-26</dc:date>
<dc:identifier>doi:10.1101/2024.03.21.586113</dc:identifier>
<dc:title><![CDATA[Sir2 and Fun30 regulate ribosomal DNA replication timing via Mcm helicase positioning and nucleosome occupancy]]></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/10.1101/2024.03.21.586169v1?rss=1">
<title>
<![CDATA[
MICROMANIPULATION OF LIVE MICRODISSECTED TISSUES WITH A LOW-COST INTEGRATED ROBOTIC PLATFORM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.21.586169v1?rss=1</link>
<description><![CDATA[
The scarcity of human biopsies available for drug testing is a paramount challenge for developing new therapeutics, disease models, and personalized treatments. Microtechnologies that combine the microscale manipulation of tissues and fluids offer the exciting possibility of miniaturizing both disease models and drug testing workflows on scarce human biopsies. Unfortunately, these technologies presently require microfluidic devices or robotic dispensers that are not widely accessible. We have rapidly-prototyped an inexpensive platform based on an off-the-shelf robot that can microfluidically manipulate live microtissues into/out of culture plates without using complicated accessories such as microscopes or pneumatic controllers. The robot integrates complex functions with a simple, cost-effective and compact construction, allowing placement inside a tissue culture hood for sterile workflows. We demonstrated a proof-of-concept cancer drug evaluation workflow of potential clinical utility using patient tumor biopsies with multiple drugs on 384-well plates. Our user-friendly, low-cost platform promises to make drug testing of microtissues broadly accessible to pharmaceutical, clinical, and biological laboratories.

TeaserA low-cost robot for handling microtissues and catalyzing their use in cancer drug evaluation and personalized oncology.
]]></description>
<dc:creator>Gottshall, N. R.</dc:creator>
<dc:creator>Stepanov, I.</dc:creator>
<dc:creator>Ahmadianyazdi, A.</dc:creator>
<dc:creator>Sinha, D.</dc:creator>
<dc:creator>Lockhart, E.</dc:creator>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Hassan, S.</dc:creator>
<dc:creator>Horowitz, L.</dc:creator>
<dc:creator>Yeung, R.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:creator>Folch, A.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.21.586169</dc:identifier>
<dc:title><![CDATA[MICROMANIPULATION OF LIVE MICRODISSECTED TISSUES WITH A LOW-COST INTEGRATED ROBOTIC PLATFORM]]></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/10.1101/2024.03.22.586189v1?rss=1">
<title>
<![CDATA[
Microdissected tumor cuboids: a microscale cancer model that retains a complex tumor microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.586189v1?rss=1</link>
<description><![CDATA[
To bridge the gap between bench and bedside, there is a need for more faithful models of human cancers that can recapitulate key features of the human tumor microenvironment (TME) and simultaneously facilitate large-scale drug tests. Our recently developed microdissection method optimizes the yield of large numbers of cuboidal microtissues ("cuboids", [~](400 {micro}m)3) from a tumor biopsy. Here we demonstrate that cuboids from syngeneic mouse tumor models and human tumors retain a complex TME, making them amenable for drug and immunotherapy evaluation. We characterize relevant TME parameters, such as cellular architecture, cytokine secretion, proteomics profiles, and response to drug panels in multi-well arrays. Despite the cutting procedure and the time spent in culture (up to 7 days), the cuboids display strong cytokine expression and drug responses, including to immunotherapy. Overall, our results suggest that cuboids could provide essential therapeutic information for personalized oncology applications and could help the development of TME-dependent therapeutics and cancer disease models, including for clinical trials.
]]></description>
<dc:creator>Horowitz, L. F.</dc:creator>
<dc:creator>Rodriguez-Mias, R. A.</dc:creator>
<dc:creator>Zhu, S.</dc:creator>
<dc:creator>Gottshall, N. R.</dc:creator>
<dc:creator>Stepanov, I.</dc:creator>
<dc:creator>Stiles, C.</dc:creator>
<dc:creator>Yeung, M.</dc:creator>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Lockhart, E. J.</dc:creator>
<dc:creator>Yeung, R. S.</dc:creator>
<dc:creator>Villen, J.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:creator>Folch, A.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.586189</dc:identifier>
<dc:title><![CDATA[Microdissected tumor cuboids: a microscale cancer model that retains a complex tumor microenvironment]]></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/10.1101/2024.03.28.587273v1?rss=1">
<title>
<![CDATA[
Carbon dioxide shapes parasite-host interactions in a human-infective nematode 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.28.587273v1?rss=1</link>
<description><![CDATA[
Skin-penetrating nematodes infect nearly one billion people worldwide. The developmentally arrested infective larvae (iL3s) seek out hosts, invade hosts via skin penetration, and resume development inside the host in a process called activation. Activated infective larvae (iL3as) traverse the host body, ending up as parasitic adults in the small intestine. Skin-penetrating nematodes respond to many chemosensory cues, but how chemosensation contributes to host seeking, intra-host development, and intra-host navigation - three crucial steps of the parasite-host interaction - remains poorly understood. Here, we investigate the role of carbon dioxide (CO2) in promoting parasite-host interactions in the human-infective threadworm Strongyloides stercoralis. We show that S. stercoralis exhibits life-stage-specific preferences for CO2: iL3s are repelled, non-infective larvae and adults are neutral, and iL3as are attracted. CO2 repulsion in iL3s may prime them for host seeking by stimulating dispersal from host feces, while CO2 attraction in iL3as may direct worms toward high-CO2 areas of the body such as the lungs and intestine. We also identify sensory neurons that detect CO2; these neurons are depolarized by CO2 in iL3s and iL3as. In addition, we demonstrate that the receptor guanylate cyclase Ss-GCY-9 is expressed specifically in CO2-sensing neurons and is required for CO2-evoked behavior. Ss-GCY-9 also promotes activation, indicating that a single receptor can mediate both behavioral and physiological responses to CO2. Our results illuminate chemosensory mechanisms that shape the interaction between parasitic nematodes and their human hosts and may aid in the design of novel anthelmintics that target the CO2-sensing pathway.
]]></description>
<dc:creator>Banerjee, N.</dc:creator>
<dc:creator>Gang, S. S.</dc:creator>
<dc:creator>Castelletto, M. L.</dc:creator>
<dc:creator>Ruiz, F.</dc:creator>
<dc:creator>Hallem, E. A.</dc:creator>
<dc:date>2024-03-31</dc:date>
<dc:identifier>doi:10.1101/2024.03.28.587273</dc:identifier>
<dc:title><![CDATA[Carbon dioxide shapes parasite-host interactions in a human-infective nematode]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.02.586657v1?rss=1">
<title>
<![CDATA[
Cas9 RNP Physiochemical Analysis for Enhanced CRISPR-AuNP Assembly and Function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.02.586657v1?rss=1</link>
<description><![CDATA[
CRISPR therapy for hematological disease has proven effective for transplant dependent beta thalassemia and sickle cell anemia, with additional disease targets in sight. The success of these therapies relies on high rates of CRISPR-induced double strand DNA breaks in hematopoietic stem and progenitor cells (HSPC). To achieve these levels, CRISPR complexes are typically delivered by electroporation ex vivo which is toxic to HSPCs. HSPCs are then cultured in stimulating conditions that promote error-prone DNA repair, requiring conditioning with chemotherapy to facilitate engraftment after reinfusion. In vivo delivery by nanocarriers of CRISPR gene editing tools has the potential to mitigate this complexity and toxicity and make this revolutionary therapy globally available. To achieve in vivo delivery, the inherent restriction factors against oligonucleotide delivery into HSPCs, that make ex vivo manipulation including electroporation and stimulation essential, must be overcome. To this end, our group developed a CRISPR carrying gold nanoparticle (CRISPR-AuNP) capable of delivering either Cas9 or Cas12a CRISPRs as ribonucleoprotein complexes (RNP) without compromising HSPC fitness. However, the most commonly used CRISPR, Cas9, demonstrated inconsistent activity in this delivery system, with lower activity relative to Cas12a. Investigation of Cas9 RNP biophysics relative to Cas12a revealed duplex RNA instability during the initial loading onto Au cores, resulting in undetectable Cas9 loading to the particle surface. Here we demonstrate preformation of RNP before loading, coupled with optimization of the loading chemistry and conditions, resulted in 39.6 {+/-} 7.0 Cas9 RNP/AuNP without compromising RNP activity in both in vitro assays and primary human HSPC. The same alterations improved Cas12a RNP/AuNP loading 10-fold over previously reported levels. To achieve particle stability, the reported polyethyleneimine outer coating was altered to include PEGylation and the resulting 2nd generation CRISPR-AuNP demonstrates favorable nanoformulation characteristics for in vivo administration, with a hydrophilic, more neutral nanoparticle surface. Direct treatment of HSPC in vitro showed 72.5 {+/-} 7.37% uptake of 2nd generation CRISPR-AuNP in primary human HSPC, but with endosomal accumulation and low rates of gene editing consistent with low levels of endosomal escape.
]]></description>
<dc:creator>Lane, D. D.</dc:creator>
<dc:creator>Gottimukkala, K. S. V.</dc:creator>
<dc:creator>Cunningham, R.</dc:creator>
<dc:creator>Cassidy, M. E.</dc:creator>
<dc:creator>Jwa, Y.</dc:creator>
<dc:creator>Castelli, J. M. P.</dc:creator>
<dc:creator>Adair, J. E.</dc:creator>
<dc:date>2024-04-02</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.586657</dc:identifier>
<dc:title><![CDATA[Cas9 RNP Physiochemical Analysis for Enhanced CRISPR-AuNP Assembly and Function]]></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/10.1101/2024.04.02.587682v1?rss=1">
<title>
<![CDATA[
Adjusting for principal components can induce spurious associations in genome-wide association studies in admixed populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.02.587682v1?rss=1</link>
<description><![CDATA[
Principal component analysis (PCA) is widely used to control for population structure in genome-wide association studies (GWAS). Top principal components (PCs) typically reflect population structure, but challenges arise in deciding how many PCs are needed and ensuring that PCs do not capture other artifacts such as regions with atypical linkage disequilibrium (LD). In response to the latter, many groups suggest performing LD pruning or excluding known high LD regions prior to PCA. However, these suggestions are not universally implemented and the implications for GWAS are not fully understood, especially in the context of admixed populations. In this paper, we investigate the impact of pre-processing and the number of PCs included in GWAS models in African American samples from the Womens Womens Health Initiative SNP Health Association Resource and two Trans-Omics for Precision Medicine Whole Genome Sequencing Project contributing studies (Jackson Heart Study and Genetic Epidemiology of Chronic Obstructive Pulmonary Disease Study). In all three samples, we find the first PC is highly correlated with genome-wide ancestry whereas later PCs often capture local genomic features. The pattern of which, and how many, genetic variants are highly correlated with individual PCs differs from what has been observed in prior studies focused on European populations and leads to distinct downstream consequences: adjusting for such PCs yields biased effect size estimates and elevated rates of spurious associations due to the phenomenon of collider bias. Excluding high LD regions identified in previous studies does not resolve these issues. LD pruning proves more effective, but the optimal choice of thresholds varies across datasets. Altogether, our work highlights unique issues that arise when using PCA to control for ancestral heterogeneity in admixed populations and demonstrates the importance of careful pre-processing and diagnostics to ensure that PCs capturing multiple local genomic features are not included in GWAS models.

Author SummaryPrincipal component analysis (PCA) is a widely used technique in human genetics research. One of its most frequent applications is in the context of genetic association studies, wherein researchers use PCA to infer, and then adjust for, the genetic ancestry of study participants. Although a powerful approach, prior work has shown that PCA sometimes captures other features or data quality issues, and pre-processing steps have been suggested to address these concerns. However, the utility and downstream implications of this recommended preprocessing are not fully understood, nor are these steps universally implemented. Moreover, the vast majority of prior work in this area was conducted in studies that exclusively included individuals of European ancestry. Here, we revisit this work in the context of admixed populations--populations with diverse, mixed ancestry that have been largely underrepresented in genetics research to date. We demonstrate the unique concerns that can arise in this context and illustrate the detrimental effects that including principal components in genetic association study models can have when not implemented carefully. Altogether, we hope our work serves as a reminder of the care that must be taken--including careful pre-processing, diagnostics, and modeling choices--when implementing PCA in admixed populations and beyond.
]]></description>
<dc:creator>Grinde, K.</dc:creator>
<dc:creator>Browning, B. L.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Thornton, T. A.</dc:creator>
<dc:creator>Browning, S. R.</dc:creator>
<dc:date>2024-04-03</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.587682</dc:identifier>
<dc:title><![CDATA[Adjusting for principal components can induce spurious associations in genome-wide association studies in admixed populations]]></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/10.1101/2024.04.05.588227v1?rss=1">
<title>
<![CDATA[
Merkel cell polyomavirus protein ALTO modulates TBK1 activity to support persistent infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.05.588227v1?rss=1</link>
<description><![CDATA[
While Merkel cell polyomavirus (MCPyV or MCV) is an abundant virus frequently shed from healthy skin, it is one of the most lethal tumor viruses in immunocompromised individuals, highlighting the crucial role of host immunity in controlling MCPyV oncogenic potential. Despite its prevalence, very little is known about how MCPyV interfaces with the host immune response to maintain asymptomatic persistent infection and how inadequate control of MCPyV infection triggers MCC tumorigenesis. In this study, we discovered that the MCPyV protein, known as the Alternative Large Tumor Open Reading Frame (ALTO), effectively primes and activates the STING signaling pathway. It recruits Src kinase into the complex of STING downstream kinase TBK1 to trigger its autophosphorylation, which ultimately activates the subsequent antiviral immune response. Combining single-cell analysis with both loss- and gain-of-function studies of MCPyV infection, we demonstrated that the activity of ALTO leads to a decrease in MCPyV replication. Thus, we have identified ALTO as a crucial viral factor that modulates the STING-TBK1 pathway, creating a negative-feedback loop that limits viral infection and maintains a delicate balance with the host immune system. Our study reveals a novel mechanism by which a tumorigenic virus-encoded protein can link Src function in cell proliferation to the activation of innate immune signaling, thereby controlling viral spread and sustaining persistent infection. Our previous findings suggest that STING also functions as a tumor suppressor in MCPyV-driven oncogenesis. This research provides a foundation for investigating how disruptions in the finely tuned virus-host balance, maintained by STING, could alter the fate of MCPyV infection, potentially encouraging malignancy.

Author summaryMerkel cell polyomavirus (MCPyV) is a small DNA virus responsible for the majority cases of Merkel Cell Carcinoma (MCC), a rare yet highly aggressive form of cancer. While MCPyV latently infects a vast majority of individuals, the mechanisms governing its control, persistence, and oncogenic triggers remain obscure. Our research reveals that the MCPyV-derived Alternative Large Tumor Open Reading Frame (ALTO) protein primes and activates the cGAS-STING- TBK1 innate immune pathway within cells. Such activation elicits an antiviral response, effectively curbing MCPyV replication. This phenomenon illustrates the viruss cunning strategy to exploit cellular mechanisms, ensuring low viral presence while facilitating long-term infection. Consequently, our study sheds light on a novel tactic utilized by MCPyV to maintain a persistent infection in its host.
]]></description>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Senay, T. E.</dc:creator>
<dc:creator>Luo, T. T.</dc:creator>
<dc:creator>Regan, J. M.</dc:creator>
<dc:creator>Salisbury, N.</dc:creator>
<dc:creator>Galloway, D.</dc:creator>
<dc:creator>You, J.</dc:creator>
<dc:date>2024-04-05</dc:date>
<dc:identifier>doi:10.1101/2024.04.05.588227</dc:identifier>
<dc:title><![CDATA[Merkel cell polyomavirus protein ALTO modulates TBK1 activity to support persistent infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.09.588340v1?rss=1">
<title>
<![CDATA[
Single Cell Analysis of Treatment-Resistant Prostate Cancer: Implications of Cell State Changes for Cell Surface Antigen Targeted Therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.09.588340v1?rss=1</link>
<description><![CDATA[
Targeting cell surface molecules using radioligand and antibody-based therapies has yielded considerable success across cancers. However, it remains unclear how the expression of putative lineage markers, particularly cell surface molecules, varies in the process of lineage plasticity, wherein tumor cells alter their identity and acquire new oncogenic properties. A notable example of lineage plasticity is the transformation of prostate adenocarcinoma (PRAD) to neuroendocrine prostate cancer (NEPC)--a growing resistance mechanism that results in the loss of responsiveness to androgen blockade and portends dismal patient survival. To understand how lineage markers vary across the evolution of lineage plasticity in prostate cancer, we applied single cell analyses to 21 human prostate tumor biopsies and two genetically engineered mouse models, together with tissue microarray analysis (TMA) on 131 tumor samples. Not only did we observe a higher degree of phenotypic heterogeneity in castrate-resistant PRAD and NEPC than previously anticipated, but also found that the expression of molecules targeted therapeutically, namely PSMA, STEAP1, STEAP2, TROP2, CEACAM5, and DLL3, varied within a subset of gene-regulatory networks (GRNs). We also noted that NEPC and small cell lung cancer (SCLC) subtypes shared a set of GRNs, indicative of conserved biologic pathways that may be exploited therapeutically across tumor types. While this extreme level of transcriptional heterogeneity, particularly in cell surface marker expression, may mitigate the durability of clinical responses to novel antigen-directed therapies, its delineation may yield signatures for patient selection in clinical trials, potentially across distinct cancer types.

SIGNIFICANCE STATEMENTTreatment of prostate cancer is rapidly evolving with several promising new drugs targeting different cell surface antigens. Selection of patients most likely to benefit from these therapies requires an understanding of how expression of these cell surface antigens varies across patients and how they change during disease progression, particularly in tumors that undergo lineage plasticity. Using immunohistochemistry and single cell mRNA sequencing, we reveal heterogeneity of cell states across a cohort of advanced disease prostate cancer patients; this heterogeneity is not captured by conventional histology-based designations of adenocarcinoma and neuroendocrine prostate cancer. We show these cell states can be identified by gene regulatory networks that could provide additional diagnostic precision based on their correlation with clinically relevant cell surface antigen expression.
]]></description>
<dc:creator>Zaidi, S.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Chan, J. M.</dc:creator>
<dc:creator>Roudier, M.</dc:creator>
<dc:creator>Zhao, J. L.</dc:creator>
<dc:creator>Gopalan, A.</dc:creator>
<dc:creator>Wadosky, K. M.</dc:creator>
<dc:creator>Patel, R. A.</dc:creator>
<dc:creator>Sayar, E.</dc:creator>
<dc:creator>Karthaus, W. R.</dc:creator>
<dc:creator>Kates, D. H.</dc:creator>
<dc:creator>Chaudhary, O.</dc:creator>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Masilionis, I.</dc:creator>
<dc:creator>Mazutis, L.</dc:creator>
<dc:creator>Chaligne, R.</dc:creator>
<dc:creator>Obradovic, A.</dc:creator>
<dc:creator>Linkov, I.</dc:creator>
<dc:creator>Barlas, A.</dc:creator>
<dc:creator>Jungbluth, A.</dc:creator>
<dc:creator>Rekhtman, N.</dc:creator>
<dc:creator>Silber, J.</dc:creator>
<dc:creator>Manova-Todorova, K.</dc:creator>
<dc:creator>Watson, P. A.</dc:creator>
<dc:creator>True, L. D.</dc:creator>
<dc:creator>Morrissey, C. M.</dc:creator>
<dc:creator>Scher, H. I.</dc:creator>
<dc:creator>Rathkopf, D.</dc:creator>
<dc:creator>Morris, M. J.</dc:creator>
<dc:creator>Goodrich, D. W.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Haffner, M.</dc:creator>
<dc:creator>Sawyers, C.</dc:creator>
<dc:date>2024-04-12</dc:date>
<dc:identifier>doi:10.1101/2024.04.09.588340</dc:identifier>
<dc:title><![CDATA[Single Cell Analysis of Treatment-Resistant Prostate Cancer: Implications of Cell State Changes for Cell Surface Antigen Targeted Therapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.09.588752v1?rss=1">
<title>
<![CDATA[
All-trans retinoic acid induces durable tumor immunity in IDH-mutant gliomas by rescuing transcriptional repression of the CRBP1-retinoic acid axis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.09.588752v1?rss=1</link>
<description><![CDATA[
Diffuse gliomas are epigenetically dysregulated, immunologically cold, and fatal tumors characterized by mutations in isocitrate dehydrogenase (IDH). Although IDH mutations yield a uniquely immunosuppressive tumor microenvironment, the regulatory mechanisms that drive the immune landscape of IDH mutant (IDHm) gliomas remain unknown. Here, we reveal that transcriptional repression of retinoic acid (RA) pathway signaling impairs both innate and adaptive immune surveillance in IDHm glioma through epigenetic silencing of retinol binding protein 1 (RBP1) and induces a profound anti-inflammatory landscape marked by loss of inflammatory cell states and infiltration of suppressive myeloid phenotypes. Restorative retinoic acid therapy in murine glioma models promotes clonal CD4+ T cell expansion and induces tumor regression in IDHm, but not IDH wildtype (IDHwt), gliomas. Our findings provide a mechanistic rationale for RA immunotherapy in IDHm glioma and is the basis for an ongoing investigator-initiated, single-center clinical trial investigating all-trans retinoic acid (ATRA) in recurrent IDHm human subjects.
]]></description>
<dc:creator>Rao, A.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Cillo, A.</dc:creator>
<dc:creator>Sussman, J. H.</dc:creator>
<dc:creator>Sandlesh, P.</dc:creator>
<dc:creator>Tarbay, A. C.</dc:creator>
<dc:creator>Mallela, A. N.</dc:creator>
<dc:creator>Cardello, C.</dc:creator>
<dc:creator>Krueger, K.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Patterson, J.</dc:creator>
<dc:creator>Akca, E.</dc:creator>
<dc:creator>Angione, A.</dc:creator>
<dc:creator>Jaman, E.</dc:creator>
<dc:creator>Kim, W. J.</dc:creator>
<dc:creator>Allen, J.</dc:creator>
<dc:creator>Venketeswaran, A.</dc:creator>
<dc:creator>Zinn, P. O.</dc:creator>
<dc:creator>Parise, R.</dc:creator>
<dc:creator>Beumer, J.</dc:creator>
<dc:creator>Duensing, A.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:creator>Ferris, R.</dc:creator>
<dc:creator>Bagley, S. J.</dc:creator>
<dc:creator>Bruno, T. C.</dc:creator>
<dc:creator>Vignali, D. A. A.</dc:creator>
<dc:creator>Agnihotri, S.</dc:creator>
<dc:creator>Amankulor, N. M.</dc:creator>
<dc:date>2024-04-13</dc:date>
<dc:identifier>doi:10.1101/2024.04.09.588752</dc:identifier>
<dc:title><![CDATA[All-trans retinoic acid induces durable tumor immunity in IDH-mutant gliomas by rescuing transcriptional repression of the CRBP1-retinoic acid axis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.11.588992v1?rss=1">
<title>
<![CDATA[
Centromeric localization of KNL2 and CENP-C proteins in plants depends on their centromere-targeting domain and DNA-binding regions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.11.588992v1?rss=1</link>
<description><![CDATA[
In eukaryotic organisms, proper chromosome segregation during cell division depends on the centromeric histone H3 (CENH3) variant. Our previous studies identified a plant CENH3 assembly factor, Kinetochore Null2 (KNL2), that possesses a centromere-targeting motif, CENPC-k, similar to the CENPC motif in CENP-C. Additionally, we have demonstrated that KNL2 can bind DNA in vitro, independent of its CENPC-k motif. Thus, the mechanism underlying the binding of KNL2 to centromeric DNA remains elusive.

Our study shows that the CENPC-k and CENPC motifs alone are not sufficient to target the centromere in N. benthamiana and A. thaliana. In-silico analysis revealed flanking DNA-binding regions near the CENPC-k and CENPC motifs, suggesting their importance in interacting with centromeric DNA. Fusion of protein fragments containing these motifs to EYFP facilitated targeting to the centromere. Deletion of DNA-binding domains reduced the centromeric localization of KNL2-C, whereas fusion of CENPC-k to the H-NS protein from E. coli targeted it to centromeres.

We conclude that targeting of KNL2 and CENP-C proteins to centromeres is dependent on the CENPC-k/CENPC motifs, and their sequence-independent DNA-binding promotes anchoring at the centromere. Understanding the targeting mechanisms of KNL2 and CENP-C may help to engineer kinetochore structure by targeting chromatin modifying proteins to centromeres.
]]></description>
<dc:creator>Yalagapati, S. P.</dc:creator>
<dc:creator>Ahmadli, U.</dc:creator>
<dc:creator>Sinha, A.</dc:creator>
<dc:creator>Kalidass, M.</dc:creator>
<dc:creator>Dabravolski, S.</dc:creator>
<dc:creator>Zuo, S.</dc:creator>
<dc:creator>Yadala, R.</dc:creator>
<dc:creator>Rutten, T.</dc:creator>
<dc:creator>Talbert, P.</dc:creator>
<dc:creator>Lermontova, I.</dc:creator>
<dc:date>2024-04-11</dc:date>
<dc:identifier>doi:10.1101/2024.04.11.588992</dc:identifier>
<dc:title><![CDATA[Centromeric localization of KNL2 and CENP-C proteins in plants depends on their centromere-targeting domain and DNA-binding regions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.16.589816v1?rss=1">
<title>
<![CDATA[
Breaking out of the cell cycle: Including quiescence in cell cycle classification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.16.589816v1?rss=1</link>
<description><![CDATA[
Single-cell transcriptomics has unveiled a vast landscape of cellular heterogeneity in which the cell cycle is a significant component. We trained a high-resolution cell cycle classifier (ccAFv2) using single cell RNA-seq (scRNA-seq) characterized human neural stem cells. The ccAFv2 classifies six cell cycle states (G1, Late G1, S, S/G2, G2/M, and M/Early G1) and a quiescent-like G0 state (Neural G0), and it incorporates a tunable parameter to filter out less certain classifications. The ccAFv2 classifier performed better than or equivalent to other state-of-the-art methods even while classifying more cell cycle states, including G0. We demonstrate that the ccAFv2 classifier effectively generalizes the S, S/G2, G2/M, and M/Early G1 states across cell types derived from all three germ layers. While the G0, G1, and Late G1 states perform well in neuroepithelial cell types, their accuracy is lower in other cell types. However, misclassifications are confined to the G0, G1, and Late G1 states. We showcased the versatility of ccAFv2 by successfully applying it to classify cells, nuclei, and spatial transcriptomics data in humans and mice, using various normalization methods and gene identifiers. We provide methods to regress the cell cycle expression patterns out of single cell or nuclei data to uncover underlying biological signals. The classifier can be used either as an R package integrated with Seurat or a PyPI package integrated with SCANPY. We proved that ccAFv2 has enhanced accuracy, flexibility, and adaptability across various experimental conditions, establishing ccAFv2 as a powerful tool for dissecting complex biological systems, unraveling cellular heterogeneity, and deciphering the molecular mechanisms by which proliferation and quiescence affect cellular processes.
]]></description>
<dc:creator>O'Connor, S. A.</dc:creator>
<dc:creator>Garcia, L.</dc:creator>
<dc:creator>Patel, A. P.</dc:creator>
<dc:creator>Hugnot, J.-P.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:creator>Plaisier, C. L.</dc:creator>
<dc:date>2024-04-20</dc:date>
<dc:identifier>doi:10.1101/2024.04.16.589816</dc:identifier>
<dc:title><![CDATA[Breaking out of the cell cycle: Including quiescence in cell cycle classification]]></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/10.1101/2024.04.17.589977v1?rss=1">
<title>
<![CDATA[
Functional and antigenic landscape of the Nipah virus receptor binding protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.17.589977v1?rss=1</link>
<description><![CDATA[
Nipah virus recurrently spills over to humans, causing fatal infections. The viral receptor-binding protein (RBP or G) attaches to host receptors and is a major target of neutralizing antibodies. Here we use deep mutational scanning to measure how all amino-acid mutations to the RBP affect cell entry, receptor binding, and escape from neutralizing antibodies. We identify functionally constrained regions of the RBP, including sites involved in oligomerization, along with mutations that differentially modulate RBP binding to its two ephrin receptors. We map escape mutations for six anti-RBP antibodies, and find that few antigenic mutations are present in natural Nipah strains. Our findings offer insights into the potential for functional and antigenic evolution of the RBP that can inform the development of antibody therapies and vaccines.
]]></description>
<dc:creator>Larsen, B. B.</dc:creator>
<dc:creator>McMahon, T.</dc:creator>
<dc:creator>Brown, J. T.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Radford, C. E.</dc:creator>
<dc:creator>Crowe, J. E.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2024-04-19</dc:date>
<dc:identifier>doi:10.1101/2024.04.17.589977</dc:identifier>
<dc:title><![CDATA[Functional and antigenic landscape of the Nipah virus receptor binding protein]]></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/10.1101/2024.04.19.590239v1?rss=1">
<title>
<![CDATA[
Antigen affinity and site of immunization dictate B cell recall responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.19.590239v1?rss=1</link>
<description><![CDATA[
Protective antibodies against HIV-1 require unusually high levels of somatic hypermutations (SHMs) introduced in germinal centers (GCs). To achieve this, a sequential vaccination approach was proposed. Using HIV-1 antibody knock-in mice with fate-mapping genes, we examined if antigen affinity affects the outcome of the B cell recall response. Compared to high affinity boost, low affinity boost resulted in decreased numbers of memory-derived B cells in secondary GCs, but with higher average SHM, indicating an affinity threshold for memory B cells to enter secondary GCs. Upon boosting local LNs, numbers of residual GC B cells increased independent on antigen affinity, while average SHM decreased. Our results demonstrate that antigen affinity and location of the boost affect the outcome of the B cell recall response. These results can help guide the design of vaccine immunogens aiming to selectively engage specific B cell clones for further SHM diversification.
]]></description>
<dc:creator>Termote, M.</dc:creator>
<dc:creator>Marques, R. C.</dc:creator>
<dc:creator>Hyllner, E.</dc:creator>
<dc:creator>Guryleva, M. V.</dc:creator>
<dc:creator>Henskens, M.</dc:creator>
<dc:creator>Brutscher, A.</dc:creator>
<dc:creator>Baken, I. J. L.</dc:creator>
<dc:creator>Castro Dopico, X.</dc:creator>
<dc:creator>Dalmau Gasull, A.</dc:creator>
<dc:creator>Murrell, B.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Westerberg, L. S.</dc:creator>
<dc:creator>Dosenovic, P.</dc:creator>
<dc:date>2024-04-23</dc:date>
<dc:identifier>doi:10.1101/2024.04.19.590239</dc:identifier>
<dc:title><![CDATA[Antigen affinity and site of immunization dictate B cell recall responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.24.590836v1?rss=1">
<title>
<![CDATA[
Delineating the functional activity of antibodies with cross-reactivity to SARS-CoV-2, SARS-CoV-1 and related sarbecoviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.24.590836v1?rss=1</link>
<description><![CDATA[
The recurring spillover of pathogenic coronaviruses and demonstrated capacity of sarbecoviruses, such SARS-CoV-2, to rapidly evolve in humans underscores the need to better understand immune responses to this virus family. For this purpose, we characterized the functional breadth and potency of antibodies targeting the receptor binding domain (RBD) of the spike glycoprotein that exhibited cross-reactivity against SARS-CoV-2 variants, SARS-CoV-1 and sarbecoviruses from diverse clades and animal origins with spillover potential. One neutralizing antibody, C68.61, showed remarkable neutralization breadth against both SARS-CoV-2 variants and viruses from different sarbecovirus clades. C68.61, which targets a conserved RBD class 5 epitope, did not select for escape variants of SARS-CoV-2 or SARS-CoV-1 in culture nor have predicted escape variants among circulating SARS-CoV-2 strains, suggesting this epitope is functionally constrained. We identified 11 additional SARS-CoV-2/SARS-CoV-1 cross-reactive antibodies that target the more sequence conserved class 4 and class 5 epitopes within RBD that show activity against a subset of diverse sarbecoviruses with one antibody binding every single sarbecovirus RBD tested. A subset of these antibodies exhibited Fc-mediated effector functions as potent as antibodies that impact infection outcome in animal models. Thus, our study identified antibodies targeting conserved regions across SARS-CoV-2 variants and sarbecoviruses that may serve as therapeutics for pandemic preparedness as well as blueprints for the design of immunogens capable of eliciting cross-neutralizing responses.

AUTHOR SUMMARYThere is a large collection of sarbecoviruses related to SARS-CoV-2 circulating in animal reservoirs with the potential to spillover into humans. Neutralizing antibodies have the potential to protect against infection, although viral escape is common. In this study, we isolated several monoclonal antibodies that show broad activity against different sarbecoviruses. The antibodies target epitopes in the core of the receptor binding domain that are highly conserved in sequence across sarbecoviruses and emerging SARS-CoV-2 variants. One antibody showed remarkable breadth against both SARS-CoV-1 variants as well as diverse sarbecoviruses. The results of deep mutational scanning suggest that mutations at these predicted sites of escape may functionally constrain viral fitness. Our functional profiling of cross-reactive antibodies highlights vulnerable sites of sarbecoviruses, with some antibodies poised as broadly neutralizing candidates for therapeutic use against future sarbecovirus emergence.
]]></description>
<dc:creator>Ruiz, F.</dc:creator>
<dc:creator>Foreman, W.</dc:creator>
<dc:creator>Lilly, M.</dc:creator>
<dc:creator>Baharani, V. A.</dc:creator>
<dc:creator>Depierreux, D. M.</dc:creator>
<dc:creator>Chohan, V.</dc:creator>
<dc:creator>Taylor, A. L.</dc:creator>
<dc:creator>Guenthoer, J.</dc:creator>
<dc:creator>Ralph, D.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Cote, M.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2024-04-25</dc:date>
<dc:identifier>doi:10.1101/2024.04.24.590836</dc:identifier>
<dc:title><![CDATA[Delineating the functional activity of antibodies with cross-reactivity to SARS-CoV-2, SARS-CoV-1 and related sarbecoviruses]]></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/10.1101/2024.04.25.591218v1?rss=1">
<title>
<![CDATA[
Cell Simulation as Cell Segmentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.25.591218v1?rss=1</link>
<description><![CDATA[
Single-cell spatial transcriptomics promises a highly detailed view of a cells transcriptional state and microenvironment, yet inaccurate cell segmentation can render this data murky by misattributing large numbers of transcripts to nearby cells or conjuring nonexistent cells. We adopt methods from ab initio cell simulation to rapidly infer morphologically plausible cell boundaries that preserve cell type heterogeneity. Benchmarking applied to datasets generated by three commercial platforms show superior performance and computational efficiency of this approach compared with existing methods. We show that improved accuracy in cell segmentation aids greatly in detection of difficult to accurately segment tumor infiltrating immune cells such as neutrophils and T cells. Lastly, through improvements in our ability to delineate subsets of tumor infiltrating T cells, we show that CXCL13-expressing CD8+ T cells tend to be more closely associated with tumor cells than their CXCL13-negative counterparts in data generated from renal cell carcinoma patient samples.
]]></description>
<dc:creator>Jones, D. C.</dc:creator>
<dc:creator>Elz, A. E.</dc:creator>
<dc:creator>Hadadianpour, A.</dc:creator>
<dc:creator>Ryu, H.</dc:creator>
<dc:creator>Glass, D. R.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:date>2024-04-28</dc:date>
<dc:identifier>doi:10.1101/2024.04.25.591218</dc:identifier>
<dc:title><![CDATA[Cell Simulation as Cell Segmentation]]></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/10.1101/2024.04.26.591029v1?rss=1">
<title>
<![CDATA[
Functional genomics screens reveal a role for TBC1D24 and SV2B in antibody-dependent enhancement of dengue virus infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.26.591029v1?rss=1</link>
<description><![CDATA[
Dengue virus (DENV) can hijack non-neutralizing IgG antibodies to facilitate its uptake into target cells expressing Fc gamma receptors (FcgR) - a process known as antibody-dependent enhancement (ADE) of infection. Beyond a requirement for FcgR, host dependency factors for this non-canonical infection route remain unknown. To identify cellular factors exclusively required for ADE, here, we performed CRISPR knockout screens in an in vitro system permissive to infection only in the presence of IgG antibodies. Validating our approach, a top hit was FcgRIIa, which facilitates binding and internalization of IgG-bound DENV but is not required for canonical infection. Additionally, we identified host factors with no previously described role in DENV infection, including TBC1D24 and SV2B, both of which have known functions in regulated secretion. Using genetic knockout and trans-complemented cells, we validated a functional requirement for these host factors in ADE assays performed with monoclonal antibodies and polyclonal sera in multiple cell lines and using all four DENV serotypes. We show that knockout of TBC1D24 or SV2B impaired binding of IgG-DENV complexes to cells without affecting FcgRIIa expression levels. Thus, we identify cellular factors beyond FcgR that are required for ADE of DENV infection. Our findings represent a first step towards advancing fundamental knowledge behind the biology of ADE that can ultimately be exploited to inform vaccination and therapeutic approaches.
]]></description>
<dc:creator>Belmont, L.</dc:creator>
<dc:creator>Contreras, M.</dc:creator>
<dc:creator>Cartwright-Acar, C. H.</dc:creator>
<dc:creator>Marceau, C. D.</dc:creator>
<dc:creator>Agrawal, A.</dc:creator>
<dc:creator>Levoir, L. M.</dc:creator>
<dc:creator>Lubow, J.</dc:creator>
<dc:creator>Goo, L.</dc:creator>
<dc:date>2024-04-27</dc:date>
<dc:identifier>doi:10.1101/2024.04.26.591029</dc:identifier>
<dc:title><![CDATA[Functional genomics screens reveal a role for TBC1D24 and SV2B in antibody-dependent enhancement of dengue virus infection]]></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/10.1101/2024.04.26.591383v1?rss=1">
<title>
<![CDATA[
Reduced Levels of Lagging Strand Polymerases Shape Stem Cell Chromatin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.26.591383v1?rss=1</link>
<description><![CDATA[
Stem cells display asymmetric histone inheritance while non-stem progenitor cells exhibit symmetric patterns in the Drosophila male germline lineage. Here, we report that components involved in lagging strand synthesis, such as DNA polymerase  and {delta} (Pol and Pol{delta}), have significantly reduced levels in stem cells compared to progenitor cells. Compromising Pol genetically induces the replication-coupled histone incorporation pattern in progenitor cells to be indistinguishable from that in stem cells, which can be recapitulated using a Pol inhibitor in a concentration-dependent manner. Furthermore, stem cell-derived chromatin fibers display a higher degree of old histone recycling by the leading strand compared to progenitor cell-derived chromatin fibers. However, upon reducing Pol levels in progenitor cells, the chromatin fibers now display asymmetric old histone recycling just like GSC-derived fibers. The old versus new histone asymmetry is comparable between stem cells and progenitor cells at both S-phase and M-phase. Together, these results indicate that developmentally programmed expression of key DNA replication components is important to shape stem cell chromatin. Furthermore, manipulating one crucial DNA replication component can induce replication-coupled histone dynamics in non-stem cells in a manner similar to that in stem cells.

One Sentence SummaryDelayed lagging strand synthesis regulates asymmetric histone incorporation.
]]></description>
<dc:creator>Snedeker, J.</dc:creator>
<dc:creator>Davis, B. E. M.</dc:creator>
<dc:creator>Ranjan, R.</dc:creator>
<dc:creator>Wooten, M. I.</dc:creator>
<dc:creator>Blundon, J.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:date>2024-04-29</dc:date>
<dc:identifier>doi:10.1101/2024.04.26.591383</dc:identifier>
<dc:title><![CDATA[Reduced Levels of Lagging Strand Polymerases Shape Stem Cell Chromatin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.02.592270v1?rss=1">
<title>
<![CDATA[
Tumor microenvironment noise-induced polarization: the main challenge in macrophages' immunotherapy for cancer. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.02.592270v1?rss=1</link>
<description><![CDATA[
Disturbance of epigenetic processes can lead to altered gene function and malignant cellular transformation. In particular, changes in the epigenetic landscape are a central topic in cancer biology. The initiation and progression of cancer are now recognized to involve both epigenetic and genetic alterations. In this paper, we study the epigenetic mechanism (related to the tumor microenvironment) responsible for increasing tumor-associated macrophages that promote the occurrence and metastasis of tumor cells, support tumor angiogenesis, inhibit T cell-mediated anti-tumor immune response, and lead to tumor progression. We show that the tumor benefits from the macrophages high degree of plasticity and larger epigenetic basins corresponding to phenotypes that favor cancer development through a process that we call noise-induced polarization. Moreover, we propose a mechanism to promote the appropriate epigenetic stability for immunotherapies involving macrophages, which includes p53 and APR-246 (eprenetapopt). Our results show that a combination therapy may be necessary to ensure the proper epigenetic stability of macrophages, which otherwise will contribute to cancer progression. On the other hand, we conclude that macrophages may remain in the anti-tumoral state in types of cancer that exhibit less TP53 mutation, like colorectal cancer; in these cases, macrophages immunotherapy may be more suitable. We finally mention the relevance of the epigenetic potential (Waddingtons landscape) as the backbone for our study, which encapsulates the biological information of the system.
]]></description>
<dc:creator>Sierra, J.</dc:creator>
<dc:creator>Avila-Ponce de Leon, U.</dc:creator>
<dc:creator>Padilla-Longoria, P.</dc:creator>
<dc:date>2024-05-05</dc:date>
<dc:identifier>doi:10.1101/2024.05.02.592270</dc:identifier>
<dc:title><![CDATA[Tumor microenvironment noise-induced polarization: the main challenge in macrophages' immunotherapy for cancer.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.05.592584v1?rss=1">
<title>
<![CDATA[
Bispecific antibodies with broad neutralization potency against SARS-CoV-2 variants of concern 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.05.592584v1?rss=1</link>
<description><![CDATA[
The ongoing emergence of SARS-CoV-2 variants of concern (VOCs) that reduce the effectiveness of antibody therapeutics necessitates development of next-generation antibody modalities that are resilient to viral evolution. Here, we characterized N-terminal domain (NTD) and receptor binding domain (RBD)-specific monoclonal antibodies previously isolated from COVID-19 convalescent donors for their activity against emergent SARS-CoV-2 VOCs. Among these, the NTD-specific antibody C1596 displayed the greatest breadth of binding to VOCs, with cryo-EM structural analysis revealing recognition of a distinct NTD epitope outside of the site i antigenic supersite. Given C1596s favorable binding profile, we designed a series of bispecific antibodies (bsAbs) termed CoV2-biRNs, that featured both NTD and RBD specificities. Notably, two of the C1596-inclusive bsAbs, CoV2-biRN5 and CoV2-biRN7, retained potent in vitro neutralization activity against all Omicron variants tested, including XBB.1.5, EG.5.1, and BA.2.86, contrasting the diminished potency of parental antibodies delivered as monotherapies or as a cocktail. Furthermore, prophylactic delivery of CoV2-biRN5 significantly reduced the viral load within the lungs of K18-hACE2 mice following challenge with SARS-CoV-2 XBB.1.5. In conclusion, our NTD-RBD bsAbs offer promising potential for the design of resilient, next-generation antibody therapeutics against SARS-CoV-2 VOCs.

One Sentence SummaryBispecific antibodies with a highly cross-reactive NTD antibody demonstrate resilience to SARS-CoV-2 variants of concern.
]]></description>
<dc:creator>Rubio, A. A.</dc:creator>
<dc:creator>Baharani, V. A.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Parada, M.</dc:creator>
<dc:creator>Abernathy, M. E.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Lee, Y. E.</dc:creator>
<dc:creator>Eso, M.</dc:creator>
<dc:creator>Phung, J.</dc:creator>
<dc:creator>Ramos, I.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>El Nesr, G.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:creator>Barnes, C. O.</dc:creator>
<dc:date>2024-05-06</dc:date>
<dc:identifier>doi:10.1101/2024.05.05.592584</dc:identifier>
<dc:title><![CDATA[Bispecific antibodies with broad neutralization potency against SARS-CoV-2 variants of concern]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-06</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/10.1101/2024.05.14.594150v1?rss=1">
<title>
<![CDATA[
EZH2-driven immune evasion defines high-risk pediatric AML with t(16;21) FUS::ERG gene fusion. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.14.594150v1?rss=1</link>
<description><![CDATA[
The past 25 years of clinical trials have produced few improvements in pediatric AML (pAML) outcomes. This is acutely evident in patients with t(16;21)(p11;q22), yielding FUS::ERG. Patients with FUS::ERG-positive AML relapse quickly and do not respond to transplantation. Major histocompatibility complex (MHC) class I & II receptors and costimulatory molecules are absent at diagnosis in FUS::ERG-positive AML, mirroring the phenotype and outcomes of post-transplant relapse. We show that this is driven by overexpression of EZH2, in vitro and in multiple clinical cohorts. While FUS::ERG AML is the most extreme example, this phenotype is shared by lethal CBFA2T3::GLIS2-driven AML, and patients with RUNX1::RUNX1T1 have significantly worse outcomes when EZH2 overexpression co-occurs. The FDA-approved EZH2 inhibitor tazemetostat reverses this phenotype, re-establishes MHC presentation, and elicits immune effector cell-mediated elimination. EZH2 inhibitors may provide the first targeted therapeutic frontline option for AML patients with FUS::ERG, with the potential for broader frontline immunostimulatory benefits.

STATEMENT OF SIGNIFICANCEHere we show an immune-evasive phenotype, present at diagnosis and characterized by elevated EZH2 levels and loss of MHC class I and II, defines a high-risk subtype of acute leukemia. Treatment with the EZH2 inhibitor tazemetostat and IFN-{gamma} reverses this phenotype and results in immune cell engagement and blast elimination.
]]></description>
<dc:creator>Buteyn, N. J.</dc:creator>
<dc:creator>Burke, C. G.</dc:creator>
<dc:creator>Sartori, V. J.</dc:creator>
<dc:creator>Deering-Gardner, E.</dc:creator>
<dc:creator>DeBruine, Z. J.</dc:creator>
<dc:creator>Kamarudin, D.</dc:creator>
<dc:creator>Chandler, D. P.</dc:creator>
<dc:creator>Monovich, A. C.</dc:creator>
<dc:creator>Perez, M. W.</dc:creator>
<dc:creator>Yi, J. S.</dc:creator>
<dc:creator>Ries, R. E.</dc:creator>
<dc:creator>Alonzo, T. A.</dc:creator>
<dc:creator>Ryan, R. J.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Triche, T. J.</dc:creator>
<dc:date>2024-05-17</dc:date>
<dc:identifier>doi:10.1101/2024.05.14.594150</dc:identifier>
<dc:title><![CDATA[EZH2-driven immune evasion defines high-risk pediatric AML with t(16;21) FUS::ERG gene fusion.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.21.595217v1?rss=1">
<title>
<![CDATA[
Apoptotic cells promote circulating tumor cell survival and metastasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.21.595217v1?rss=1</link>
<description><![CDATA[
During tumor progression and especially following cytotoxic therapy, cell death of both tumor and stromal cells is widespread. Despite clinical observations that high levels of apoptotic cells correlate with poorer patient outcomes, the physiological effects of dying cells on tumor progression remain incompletely understood. Here, we report that circulating apoptotic cells robustly enhance tumor cell metastasis to the lungs. Using intravenous metastasis models, we observed that the presence of apoptotic cells, but not cells dying by other mechanisms, supports circulating tumor cell (CTC) survival following arrest in the lung vasculature. Apoptotic cells promote CTC survival by recruiting platelets to the forming metastatic niche. Apoptotic cells externalize the phospholipid phosphatidylserine to the outer leaflet of the plasma membrane, which we found increased the activity of the coagulation initiator Tissue Factor, thereby triggering the formation of platelet clots that protect proximal CTCs. Inhibiting the ability of apoptotic cells to induce coagulation by knocking out Tissue Factor, blocking phosphatidylserine, or administering the anticoagulant heparin abrogated the pro-metastatic effect of apoptotic cells. This work demonstrates a previously unappreciated role for apoptotic cells in facilitating metastasis by establishing CTC-supportive emboli, and suggests points of intervention that may reduce the pro-metastatic effect of apoptotic cells.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/595217v1_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@889460org.highwire.dtl.DTLVardef@c55e30org.highwire.dtl.DTLVardef@1aac38forg.highwire.dtl.DTLVardef@cf7ae_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Hagan, C. E.</dc:creator>
<dc:creator>Snyder, A. G.</dc:creator>
<dc:creator>Headley, M.</dc:creator>
<dc:creator>Oberst, A.</dc:creator>
<dc:date>2024-05-22</dc:date>
<dc:identifier>doi:10.1101/2024.05.21.595217</dc:identifier>
<dc:title><![CDATA[Apoptotic cells promote circulating tumor cell survival and metastasis]]></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/10.1101/2024.05.22.595399v1?rss=1">
<title>
<![CDATA[
A Rigorous Multi-Laboratory Study of Known PDAC Biomarkers Identifies Increased Sensitivity and Specificity Over CA19-9 Alone 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.22.595399v1?rss=1</link>
<description><![CDATA[
A blood test that enables surveillance for early-stage pancreatic ductal adenocarcinoma (PDAC) is an urgent need. Independent laboratories have reported PDAC biomarkers that could improve biomarker performance over CA19-9 alone, but the performance of the previously reported biomarkers in combination is not known. Therefore, we conducted a coordinated case/control study across multiple laboratories using common sets of blinded training and validation samples (132 and 295 plasma samples, respectively) from PDAC patients and non-PDAC control subjects representing conditions under which surveillance occurs. We analyzed the training set to identify candidate biomarker combination panels using biomarkers across laboratories, and we applied the fixed panels to the validation set. The panels identified in the training set, CA19-9 with CA199.STRA, LRG1, TIMP-1, TGM2, THSP2, ANG, and MUC16.STRA, achieved consistent performance in the validation set. The panel of CA19-9 with the glycan biomarker CA199.STRA improved sensitivity from 0.44 with 0.98 specificity for CA19-9 alone to 0.71 with 0.98 specificity (p < 0.001, 1000-fold bootstrap). Similarly, CA19-9 combined with the protein biomarker LRG1 and CA199.STRA improved specificity from 0.16 with 0.94 sensitivity for CA19-9 to 0.65 with 0.89 sensitivity (p < 0.001, 1000-fold bootstrap). We further validated significantly improved performance using biomarker panels that did not include CA19-9. This study establishes the effectiveness of a coordinated study of previously discovered biomarkers and identified panels of those biomarkers that significantly increased the sensitivity and specificity of early-stage PDAC detection in a rigorous validation trial.
]]></description>
<dc:creator>Haab, B.</dc:creator>
<dc:creator>Qian, L.</dc:creator>
<dc:creator>Staal, B.</dc:creator>
<dc:creator>Jain, M.</dc:creator>
<dc:creator>Fahrmann, J. F.</dc:creator>
<dc:creator>Worthington, C.</dc:creator>
<dc:creator>Prosser, D.</dc:creator>
<dc:creator>Velikokhatnaya, L.</dc:creator>
<dc:creator>Lopez, C.</dc:creator>
<dc:creator>Tang, R.</dc:creator>
<dc:creator>Hurd, M.</dc:creator>
<dc:creator>Gopalakrishnan, N.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Smith, L.</dc:creator>
<dc:creator>Hanash, S.</dc:creator>
<dc:creator>Batra, S. K.</dc:creator>
<dc:creator>Maitra, A.</dc:creator>
<dc:creator>Lokshin, A.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Brand, R.</dc:creator>
<dc:date>2024-05-26</dc:date>
<dc:identifier>doi:10.1101/2024.05.22.595399</dc:identifier>
<dc:title><![CDATA[A Rigorous Multi-Laboratory Study of Known PDAC Biomarkers Identifies Increased Sensitivity and Specificity Over CA19-9 Alone]]></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/10.1101/2024.05.24.595249v1?rss=1">
<title>
<![CDATA[
Multiplexed single cell transcriptomics optimizes mesodermal patterning and hemogenic endothelial output from murine embryonic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.24.595249v1?rss=1</link>
<description><![CDATA[
Early patterning of mesodermal precursor populations is a key step of hematopoietic development in the embryo. To better understand this process, we employed sci-Plex, a high-throughput method of measuring multiplexed perturbations at the single-cell level, to evaluate the transcriptional response of mouse embryonic stem cells subjected to a gradient of two key morphogens in early mesoderm/hematopoietic development, Activin and BMP4. sci-Plex revealed varying combinations of Activin and BMP4 temporally influenced mesoderm patterning in vitro and subsequent production of cell types reflecting their in vivo counterparts. We leveraged sci-Plex data to further optimize the generation of intraembryonic-like hemogenic endothelial cells that serve as the precursors of hematopoietic lineages, including hematopoietic stem cells. This study highlights the utility of sci-Plex to dissect how dose and temporal integration of interacting signal pathways determines cell fates and serves as a resource to analyze cell fate choices in early mesoderm patterning at single cell resolution.
]]></description>
<dc:creator>Varnum-Finney, B.</dc:creator>
<dc:creator>Heck, A. M.</dc:creator>
<dc:creator>Srivatsan, S.</dc:creator>
<dc:creator>Dozono, S.</dc:creator>
<dc:creator>Wellington, R.</dc:creator>
<dc:creator>Nourigat-McKay, C.</dc:creator>
<dc:creator>Dignum, T.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Hadland, B.</dc:creator>
<dc:date>2024-05-26</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.595249</dc:identifier>
<dc:title><![CDATA[Multiplexed single cell transcriptomics optimizes mesodermal patterning and hemogenic endothelial output from murine embryonic stem cells]]></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/10.1101/2024.05.30.596749v1?rss=1">
<title>
<![CDATA[
Under pressure: altered endothelial flow response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.30.596749v1?rss=1</link>
<description><![CDATA[
Blood flow within the vasculature is a critical determinant of endothelial cell (EC) identity and functionality, yet the intricate interplay of various hemodynamic forces and their collective impact on endothelial and vascular responses are not fully understood. Specifically, the role of hydrostatic pressure in the EC flow response is understudied, despite its known significance in vascular development and disease. To address this gap, we developed in vitro models to investigate how pressure influences EC responses to flow. Our study demonstrates that elevated pressure conditions significantly modify shear-induced flow alignment and increase endothelial cell density. Bulk and single-cell RNA sequencing analyses revealed that, while shear stress remains the primary driver of flow-induced transcriptional changes, pressure modulates shear- induced signaling in a dose-dependent manner. These pressure-responsive transcriptional signatures identified in human ECs were conserved during the onset of circulation in early mouse embryonic vascular development, where pressure was notably associated with transcriptional programs essential to arterial and hemogenic EC fates. Our findings suggest that pressure plays a synergistic role with shear stress on ECs and emphasizes the need for an integrative approach to endothelial cell mechanotransduction, one that encompasses the effects induced by pressure alongside other hemodynamic forces.
]]></description>
<dc:creator>Mandrycky, C.</dc:creator>
<dc:creator>Ishida, T.</dc:creator>
<dc:creator>Rayner, S. G.</dc:creator>
<dc:creator>Heck, A. M.</dc:creator>
<dc:creator>Hadland, B.</dc:creator>
<dc:creator>ZHENG, Y.</dc:creator>
<dc:date>2024-06-02</dc:date>
<dc:identifier>doi:10.1101/2024.05.30.596749</dc:identifier>
<dc:title><![CDATA[Under pressure: altered endothelial flow response]]></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/10.1101/2024.06.03.597178v1?rss=1">
<title>
<![CDATA[
Hallmarks of tumor-experienced T cells are absent in multiple myeloma patients from diagnosis through maintenance therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.03.597178v1?rss=1</link>
<description><![CDATA[
Dysregulation of the bone marrow (BM) niche in multiple myeloma (MM) alters the composition and state of resident immune cells, potentially impeding anti-tumor immunity. One common mechanism of immune inhibition in solid tumors is the induction of exhaustion in tumor-specific T cells. However, the extent of T cell tumor recognition and exhaustion is not well-characterized in MM. As the specific mechanisms of immune evasion are critical for devising effective therapeutic strategies, we deeply profiled the CD8+ T cell compartment of newly-diagnosed MM (NDMM) patients for evidence of tumor reactivity and T cell exhaustion. We applied single-cell multi-omic sequencing and antigen-specific mass cytometry to longitudinal BM and peripheral blood (PB) samples taken from timepoints spanning from diagnosis through induction therapy, autologous stem cell transplant (ASCT), and maintenance therapy. We identified an exhausted-like population that lacked several canonical exhaustion markers, was not significantly enriched in NDMM patients, and consisted of small, nonpersistent clones. We also observed an activated population with increased frequency in the PB of NDMM patients exhibiting phenotypic and clonal features consistent with homeostatic, antigen-nonspecific activation. However, there was no evidence of "tumor-experienced" T cells displaying hallmarks of terminal exhaustion and/or tumor-specific activation/expansion in NDMM patients at any timepoint.
]]></description>
<dc:creator>Shasha, C.</dc:creator>
<dc:creator>Glass, D. R.</dc:creator>
<dc:creator>Moelhman, E.</dc:creator>
<dc:creator>Islas, L.</dc:creator>
<dc:creator>Tian, Y.</dc:creator>
<dc:creator>Szeto, G. L.</dc:creator>
<dc:creator>Peng, T.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Wurscher, M.</dc:creator>
<dc:creator>Bumol, T. F.</dc:creator>
<dc:creator>Torgerson, T. R.</dc:creator>
<dc:creator>Greenberg, P. D.</dc:creator>
<dc:creator>Green, D. J.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:date>2024-06-04</dc:date>
<dc:identifier>doi:10.1101/2024.06.03.597178</dc:identifier>
<dc:title><![CDATA[Hallmarks of tumor-experienced T cells are absent in multiple myeloma patients from diagnosis through maintenance therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.10.598377v1?rss=1">
<title>
<![CDATA[
Converging cytokine and metabolite networks shape asymmetric T cell fate at the term human maternal-fetal interface 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.10.598377v1?rss=1</link>
<description><![CDATA[
Placentation presents immune conflict between mother and fetus, yet in normal pregnancy maternal immunity against infection is maintained without expense to fetal tolerance. This is believed to result from adaptations at the maternal-fetal interface (MFI) which affect T cell programming, but the identities (i.e., memory subsets and antigenic specificities) of T cells and the signals that mediate T cell fates and functions at the MFI remain poorly understood. We found intact recruitment programs as well as pro-inflammatory cytokine networks that can act on maternal T cells in an antigen-independent manner. These inflammatory signals elicit T cell expression of co-stimulatory receptors necessary for tissue retention, which can be engaged by local macrophages. Although pro-inflammatory molecules elicit T cell effector functions, we show that additional cytokine (TGF-{beta}1) and metabolite (kynurenine) networks may converge to tune T cell function to those of sentinels. Together, we demonstrate an additional facet of fetal tolerance, wherein T cells are broadly recruited and restrained in an antigen-independent, cytokine/metabolite-dependent manner. These mechanisms provide insight into antigen-nonspecific T cell regulation, especially in tissue microenvironments where they are enriched.
]]></description>
<dc:creator>Maurice, N. J.</dc:creator>
<dc:creator>Erickson, J. R.</dc:creator>
<dc:creator>DeJong, C. S.</dc:creator>
<dc:creator>Mair, F.</dc:creator>
<dc:creator>Taber, A. K.</dc:creator>
<dc:creator>Frutoso, M.</dc:creator>
<dc:creator>Islas, L. V.</dc:creator>
<dc:creator>Vigil, A.-L. B.</dc:creator>
<dc:creator>Lawler, R. L.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:creator>Shree, R.</dc:creator>
<dc:creator>McCartney, S. A.</dc:creator>
<dc:date>2024-06-12</dc:date>
<dc:identifier>doi:10.1101/2024.06.10.598377</dc:identifier>
<dc:title><![CDATA[Converging cytokine and metabolite networks shape asymmetric T cell fate at the term human maternal-fetal interface]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.12.598706v1?rss=1">
<title>
<![CDATA[
Prevalent chromosome fusion in Vibrio cholerae O1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.12.598706v1?rss=1</link>
<description><![CDATA[
Two circular chromosomes are a defining feature of the family Vibrionaceae, including the pathogen Vibrio cholerae, with rare reports of isolates with a single, fused chromosome. Here we report chromosome fusions in clinical V. cholerae O1 isolates, including several independent fusion events stable enough to be transmitted between patients within a household. Fusion occurs in a 12 kilobase-pair homologous sequence shared between the two chromosomes, which may lead to reversible chromosomal fusion.
]]></description>
<dc:creator>Cuenod, A.</dc:creator>
<dc:creator>Chac, D.</dc:creator>
<dc:creator>Khan, A. I.</dc:creator>
<dc:creator>Chowdhury, F.</dc:creator>
<dc:creator>Hyppa, R. W.</dc:creator>
<dc:creator>Markiewicz, S. M.</dc:creator>
<dc:creator>Calderwood, S. B.</dc:creator>
<dc:creator>Ryan, E. T.</dc:creator>
<dc:creator>Harris, J. B.</dc:creator>
<dc:creator>LaRocque, R. C.</dc:creator>
<dc:creator>Bhuiyan, T. R.</dc:creator>
<dc:creator>Smith, G. R.</dc:creator>
<dc:creator>Qadri, F.</dc:creator>
<dc:creator>Lypaczewski, P.</dc:creator>
<dc:creator>Weil, A. A.</dc:creator>
<dc:creator>Shapiro, B. J.</dc:creator>
<dc:date>2024-06-12</dc:date>
<dc:identifier>doi:10.1101/2024.06.12.598706</dc:identifier>
<dc:title><![CDATA[Prevalent chromosome fusion in Vibrio cholerae O1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.13.598910v1?rss=1">
<title>
<![CDATA[
Human cytomegalovirus induces neuronal gene expression for viral maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.13.598910v1?rss=1</link>
<description><![CDATA[
Viral invasion of the host cell causes some of the most dramatic changes in biology. Human cytomegalovirus (HCMV) extensively remodels host cells, altering nuclear shape and generating a cytoplasmic viral-induced assembly compartment (vIAC). How these striking morphology changes take place in the context of host gene regulation is still emerging. Here, we discovered that histone variant macroH2A1 is essential for producing infectious progeny. Because virion maturation and cellular remodeling are closely linked processes, we investigated structural changes in the host cell upon HCMV infection. We discovered that macroH2A1 is necessary for HCMV-induced reorganization of the host nucleus, cytoskeleton, and endoplasmic reticulum. Furthermore, using RNA-seq we found that while all viral genes were highly expressed in the absence of macroH2A1, many HCMV-induced host genes were not. Remarkably, hundreds of these HCMV-induced macroH2A1-dependent host genes are associated with neuronal synapse formation and vesicle trafficking. Knock-down of these HCMV-induced neuronal genes during infection resulted in malformed vIACs and smaller plaques, establishing their importance to HCMV infection. Together, our findings demonstrate that HCMV manipulates host gene expression by hijacking a dormant neuronal secretory pathway for efficient virion maturation.
]]></description>
<dc:creator>Kelnhofer-Millevolte, L. E.</dc:creator>
<dc:creator>Smith, J. R.</dc:creator>
<dc:creator>Nguyen, D. H.</dc:creator>
<dc:creator>Wilson, L. S.</dc:creator>
<dc:creator>Lewis, H. C.</dc:creator>
<dc:creator>Arnold, E. A.</dc:creator>
<dc:creator>Brinkley, M. R.</dc:creator>
<dc:creator>Geballe, A. P.</dc:creator>
<dc:creator>Ramachandran, S.</dc:creator>
<dc:creator>Avgousti, D. C.</dc:creator>
<dc:date>2024-06-13</dc:date>
<dc:identifier>doi:10.1101/2024.06.13.598910</dc:identifier>
<dc:title><![CDATA[Human cytomegalovirus induces neuronal gene expression for viral maturation]]></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/10.1101/2024.06.14.598510v1?rss=1">
<title>
<![CDATA[
Proteogenomic Reprogramming to a Functional Human Totipotent Stem Cell State via a PARP-DUX4 Regulatory Axis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.14.598510v1?rss=1</link>
<description><![CDATA[
PARP1 (ARTD1) and Tankyrases (TNKS1/TNKS2; PARP5a/5b) are poly-ADP-ribose polymerases (PARPs) with catalytic and non-catalytic functions that regulate both the genome and proteome during zygotic genome activation (ZGA), totipotent, and pluripotent embryonic stages. Here, we show that primed, conventional human pluripotent stem cells (hPSC) cultured continuously under non-specific TNKS1/TNKS2/PARP1-inhibited chemical naive reversion conditions underwent epigenetic reprogramming to clonal blastomere-like stem cells. TIRN stem cells (TIRN-SC) concurrently expressed hundreds of gene targets of the ZGA-priming pioneer factor DUX4, as well as a panoply of four-cell (4C)-specific (e.g., TPRXL, HOX clusters), eight-cell (8C)-specific (e.g., DUXA, GSC, GATA6), primitive endoderm-specific (e.g., GATA4, SOX17), trophectoderm-specific (e.g., CDX2, TFAP2C), and naive epiblast-specific (e.g., DNMT3L, NANOG, POU5F1(OCT4)) factors; all in a hybrid, combinatorial single-cell manner. Mapping of proteomic and single-cell expressions of TIRN-SC against human preimplantation embryo references identified them as relatively homogenous 4C-8C stage populations. Injection of TIRN cells into murine 8C-16C-staged embryos resulted in efficient totipotent-like single cell contributions of human cells to both extra-embryonic (trophectoderm, placenta) and embryonic (neural, fetal liver, hematopoietic) lineages in human-murine blastocyst and fetal chimeras. Pairing of proteome with ubiquitinome analyses of TIRN-SC revealed a global shutdown of ADP-ribosylation, and a perturbed TNKS/PARP1 equilibrium which not only impacted the protein levels of hundreds of TNKS/PARP1 substrates via a rewiring of the ubiquitin-proteosome system (UPS), but also de-repressed expression of hundreds of developmental genes associated with PARP1 suppression. ChIP-Seq analysis of core NANOG-SOX2-OCT4 (NSO) pluripotency factors in TIRN-SC identified reprogrammed DUX4-accessible distal and cis-regulatory enhancer regions that were co-bound by PARP1 (NSOP). These NSOP enhancer regions possessed co-binding motifs for hundreds of the same ZGA-associated, embryonic, and extraembryonic lineage-specifying pioneer factors (e.g., HOX, FOX, GATA, SOX, TBX, CDX families) that were concurrently co-expressed in TIRN-SC; suggesting that PARP1 and DUX4 cooperate with NSO pluripotency core factors to regulate the epigenetic plasticity of a human totipotency program. These findings provide the first demonstration that global, proteome-wide perturbations of post-translational modifications (i.e., ADP-ribosylation, ubiquitination) can regulate epigenetic reprogramming during human embryogenesis. Totipotent TIRN-SC will provide a valuable cell culture model for studying the proteogenomic regulation of lineage specification from human blastomere stages and may facilitate the efficient generation of human organs in interspecies chimeras.
]]></description>
<dc:creator>Zimmerlin, L.</dc:creator>
<dc:creator>Angarita, A.</dc:creator>
<dc:creator>Park, T. S.</dc:creator>
<dc:creator>Evans-Moses, R.</dc:creator>
<dc:creator>Thomas, J.</dc:creator>
<dc:creator>Yan, S.</dc:creator>
<dc:creator>Uribe, I.</dc:creator>
<dc:creator>Vegas, I.</dc:creator>
<dc:creator>Kochendoerfer, C.</dc:creator>
<dc:creator>Buys, W.</dc:creator>
<dc:creator>Leung, A.</dc:creator>
<dc:creator>Zambidis, E. T.</dc:creator>
<dc:date>2024-06-15</dc:date>
<dc:identifier>doi:10.1101/2024.06.14.598510</dc:identifier>
<dc:title><![CDATA[Proteogenomic Reprogramming to a Functional Human Totipotent Stem Cell State via a PARP-DUX4 Regulatory Axis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.14.599073v1?rss=1">
<title>
<![CDATA[
Weighted variance component test for the integrative multi-omics analysis of microbiome data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.14.599073v1?rss=1</link>
<description><![CDATA[
Metabolic dysregulation and alterations have been linked to various diseases and conditions. Innovations in high-throughput technology now allow rapid profiling of the metabolome and metagenome -- often the gene content of bacterial populations -- for characterizing metabolism. Due to the small sample sizes and high dimensionality of the data, pathway analysis (wherein the effect of multiple genes or metabolites on an outcome is cumulatively assessed) of metabolomic data is commonly conducted and also represents a standard for metagenomic analysis. However, how to integrate both data types remains unclear. Recognizing that a metabolic pathway can be complementarily characterized by both metagenomics and metabolomics, we propose a weighted variance components framework to test if the joint effect of genes and metabolites in a biological pathway is associated with outcomes. The approach allows analytic p-value calculation, correlation between data types, and optimal weighting. Power simulations show that our approach often outperforms other strategies while maintaining type I error. The approach is illustrated on real data.
]]></description>
<dc:creator>Zhang, A.</dc:creator>
<dc:creator>Ling, W.</dc:creator>
<dc:creator>Little, A.</dc:creator>
<dc:creator>Williams-Nguyen, J. S.</dc:creator>
<dc:creator>Moon, J.-Y.</dc:creator>
<dc:creator>Burk, R. D.</dc:creator>
<dc:creator>Knight, R.</dc:creator>
<dc:creator>Wang, D. D.</dc:creator>
<dc:creator>Qi, Q.</dc:creator>
<dc:creator>Kaplan, R.</dc:creator>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:date>2024-06-17</dc:date>
<dc:identifier>doi:10.1101/2024.06.14.599073</dc:identifier>
<dc:title><![CDATA[Weighted variance component test for the integrative multi-omics analysis of microbiome data]]></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/10.1101/2024.07.09.602730v1?rss=1">
<title>
<![CDATA[
RBM10 loss induces aberrant splicing of cytoskeletal and extracellular matrix mRNAs and promotes metastatic fitness. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.09.602730v1?rss=1</link>
<description><![CDATA[
RBM10 modulates transcriptome-wide cassette exon splicing. Loss-of-function RBM10 mutations are enriched in thyroid cancers with distant metastases. Analysis of transcriptomes and genes mis-spliced by RBM10 loss showed pro-migratory and RHO/RAC signaling signatures. RBM10 loss increases cell velocity. Cytoskeletal and ECM transcripts subject to exon-inclusion events included vinculin (VCL), tenascin C (TNC) and CD44. Knockdown of the VCL exon inclusion transcript in RBM10-null cells reduced cell velocity, whereas knockdown of TNC and CD44 exon-inclusion isoforms reduced invasiveness. RAC1-GTP levels were increased in RBM10-null cells. Mouse HrasG12V/Rbm1OKO thyrocytes develop metastases that are reversed by RBM10 or by combined knockdown of VCL, CD44 and TNC inclusion isoforms. Thus, RBM10 loss generates exon inclusions in transcripts regulating ECM-cytoskeletal interactions, leading to RAC1 activation and metastatic competency. Moreover, a CRISPR-Cas9 screen for synthetic lethality with RBM10 loss identified NFkB effectors as central to viability, providing a therapeutic target for these lethal thyroid cancers.

SUMMARYRNA splicing factor mutations are common in cancer but connecting phenotypes to specific misspliced genes has been challenging. We show that RBM10 loss leads to exon inclusions of transcripts regulating ECM-cytoskeletal interactions and RAC1-GTP activation, sufficient to promote metastatic fitness.
]]></description>
<dc:creator>Krishnamoorthy, G. P.</dc:creator>
<dc:creator>Glover, A. R.</dc:creator>
<dc:creator>Untch, B. R.</dc:creator>
<dc:creator>Sigcha-Coello, N. F.</dc:creator>
<dc:creator>Xu, B.</dc:creator>
<dc:creator>Vukel, D.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Tiedje, V.</dc:creator>
<dc:creator>Berman, K.</dc:creator>
<dc:creator>Tamarapu, P. P.</dc:creator>
<dc:creator>Acuna-Ruiz, A.</dc:creator>
<dc:creator>Saqcena, M.</dc:creator>
<dc:creator>de Stanchina, E.</dc:creator>
<dc:creator>Boucai, L.</dc:creator>
<dc:creator>Ghossein, R. A.</dc:creator>
<dc:creator>Knauf, J. A.</dc:creator>
<dc:creator>Abdel-Wahab, O.</dc:creator>
<dc:creator>Bradley, R.</dc:creator>
<dc:creator>Fagin, J. A.</dc:creator>
<dc:date>2024-07-10</dc:date>
<dc:identifier>doi:10.1101/2024.07.09.602730</dc:identifier>
<dc:title><![CDATA[RBM10 loss induces aberrant splicing of cytoskeletal and extracellular matrix mRNAs and promotes metastatic fitness.]]></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/10.1101/2024.07.11.603080v1?rss=1">
<title>
<![CDATA[
Substrate specificity and protein stability drive the divergence of plant-specific DNA methyltransferases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.11.603080v1?rss=1</link>
<description><![CDATA[
DNA methylation is an important epigenetic mechanism essential for transposon silencing and genome integrity. Across evolution, the substrates of DNA methylation have diversified between kingdoms to account for genome complexity. In plants, Chromomethylase3 (CMT3) and CMT2 are the major methyltransferases mediating CHG and CHH methylation, respectively. However, how these two enzymes diverge on substrate specificities during evolution remains unknown. Here, we reveal that CMT2 originates from a duplication of the evolutionarily more ancient CMT3 in flowering plants. Lacking a key arginine residue recognizing CHG in CMT2 impairs its CHG methylation activity in most flowering plants. An engineered V1200R mutation empowers CMT2 to restore both CHG and CHH methylation in Arabidopsis cmt2cmt3 mutant, testifying a loss-of-function effect for CMT2 after [~]200 million years of evolution. Interestingly, CMT2 has evolved a long and unstructured N-terminus critical for balancing protein stability, especially under heat stress. Furthermore, CMT2 N-terminus is plastic and can be tolerant to various natural mutations. Together, this study reveals the mechanism of chromomethylase divergence for context-specific DNA methylation in plants and sheds important lights on DNA methylation evolution and function.
]]></description>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Gwee, J.</dc:creator>
<dc:creator>Fang, J.</dc:creator>
<dc:creator>Leichter, S. M.</dc:creator>
<dc:creator>Sanders, D.</dc:creator>
<dc:creator>Ji, X.</dc:creator>
<dc:creator>Song, J.</dc:creator>
<dc:creator>Zhong, X.</dc:creator>
<dc:date>2024-07-16</dc:date>
<dc:identifier>doi:10.1101/2024.07.11.603080</dc:identifier>
<dc:title><![CDATA[Substrate specificity and protein stability drive the divergence of plant-specific DNA methyltransferases]]></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/10.1101/2024.07.16.603579v1?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/10.1101/2024.07.16.603734v1?rss=1">
<title>
<![CDATA[
Molecular consequences of acute versus chronic CDK12 loss in prostate carcinoma nominates distinct therapeutic strategies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.16.603734v1?rss=1</link>
<description><![CDATA[
Genomic loss of the transcriptional kinase CDK12 occurs in [~]6% of metastatic castration-resistant prostate cancers (mCRPC) and correlates with poor patient outcomes. Prior studies demonstrate that acute CDK12 loss confers a homologous recombination (HR) deficiency (HRd) phenotype via premature intronic polyadenylation (IPA) of key HR pathway genes, including ATM. However, mCRPC patients have not demonstrated benefit from therapies that exploit HRd such as inhibitors of polyADP ribose polymerase (PARP). Based on this discordance, we sought to test the hypothesis that an HRd phenotype is primarily a consequence of acute CDK12 loss and the effect is greatly diminished in prostate cancers adapted to CDK12 loss. Analyses of whole genome sequences (WGS) and RNA sequences (RNAseq) of human mCRPCs determined that tumors with biallelic CDK12 alterations (CDK12BAL) lack genomic scar signatures indicative of HRd, despite carrying bi-allelic loss and the appearance of the hallmark tandem-duplicator phenotype (TDP). Experiments confirmed that acute CDK12 inhibition resulted in aberrant polyadenylation and downregulation of long genes (including BRCA1 and BRCA2) but such effects were modest or absent in tumors adapted to chronic CDK12BAL. One key exception was ATM, which did retain transcript shortening and reduced protein expression in the adapted CDK12BAL models. However, CDK12BALcells demonstrated intact HR as measured by RAD51 foci formation following irradiation. CDK12BAL cells showed a vulnerability to targeting of CDK13 by sgRNA or CDK12/13 inhibitors and in vivo treatment of prostate cancer xenograft lines showed that tumors with CDK12BALresponded to the CDK12/13 inhibitor SR4835, while CDK12-intact lines did not. Collectively, these studies show that aberrant polyadenylation and long HR gene downregulation is primarily a consequence of acute CDK12 deficiency, which is largely compensated for in cells that have adapted to CDK12 loss. These results provide an explanation for why PARPi monotherapy has thus far failed to consistently benefit patients with CDK12 alterations, though alternate therapies that target CDK13 or transcription are candidates for future research and testing.
]]></description>
<dc:creator>Frank, S.</dc:creator>
<dc:creator>Persse, T.</dc:creator>
<dc:creator>Coleman, I. M.</dc:creator>
<dc:creator>Bankhead, A.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>DeSarkar, N.</dc:creator>
<dc:creator>Wilson, D.</dc:creator>
<dc:creator>Rudoy, D.</dc:creator>
<dc:creator>Vashisth, M.</dc:creator>
<dc:creator>Galipeau, P.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Hanratty, B.</dc:creator>
<dc:creator>Dumpit, R.</dc:creator>
<dc:creator>Morrissey, c.</dc:creator>
<dc:creator>Corey, E.</dc:creator>
<dc:creator>Montgomery, R.</dc:creator>
<dc:creator>Haffner, M.</dc:creator>
<dc:creator>Pritchard, C.</dc:creator>
<dc:creator>Vasioukhin, V.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:date>2024-07-19</dc:date>
<dc:identifier>doi:10.1101/2024.07.16.603734</dc:identifier>
<dc:title><![CDATA[Molecular consequences of acute versus chronic CDK12 loss in prostate carcinoma nominates distinct therapeutic strategies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.19.604135v1?rss=1">
<title>
<![CDATA[
Multiple dermal cell types support productive infection and dynamic translocation of infectious Ebola virus to the apical surface of human skin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.19.604135v1?rss=1</link>
<description><![CDATA[
Ebola virus (EBOV) within the Filoviridae family causes severe human disease. At late stages of infection, EBOV virions are found on the surface of patients skin; however, the permissive cell types within the skin and how infectious virus translocates to the apical skin surfaces is not known. Here, we describe a human transwell skin explant culture model and show that EBOV infection of human skin tissues via the basal media results in a time- and dose-dependent increase in infectious virus in dermal and epidermal tissue. Infectious virus was detected on the apical epidermal surface within 3 days, indicating that the virus propagates within and traffics through the tissue. In the dermis, EBOV-infected cells were of myeloid, endothelial and fibroblast origins, whereas keratinocytes harbored virus in the epidermis. Complementary studies showed that both purified skin fibroblasts and keratinocytes supported EBOV infection ex vivo and that both cell types required the phosphatidylserine receptor, Axl, and the endosomal protein, NPC1, for virus entry. Our experimental platform identified new susceptible cell types and demonstrated dynamic trafficking of EBOV virions that resulted in infectious virus on the skin surface; findings that may explain person-to-person transmission via skin contact.

TeaserUsing a human skin explant model, these studies identify and characterize skin cell populations that support Ebola virus infection.
]]></description>
<dc:creator>Messingham, K.</dc:creator>
<dc:creator>Richards, P. T.</dc:creator>
<dc:creator>Fleck, A.</dc:creator>
<dc:creator>Patel, R. A.</dc:creator>
<dc:creator>Djurkovic, M.</dc:creator>
<dc:creator>Elliff, J.</dc:creator>
<dc:creator>Connell, S.</dc:creator>
<dc:creator>Gonzalez, J. P. M.</dc:creator>
<dc:creator>Gourronc, F. A.</dc:creator>
<dc:creator>Dillard, J. A.</dc:creator>
<dc:creator>Davey, R. A.</dc:creator>
<dc:creator>Klingelhutz, A. J.</dc:creator>
<dc:creator>Shtanko, O.</dc:creator>
<dc:creator>Maury, W.</dc:creator>
<dc:date>2024-07-19</dc:date>
<dc:identifier>doi:10.1101/2024.07.19.604135</dc:identifier>
<dc:title><![CDATA[Multiple dermal cell types support productive infection and dynamic translocation of infectious Ebola virus to the apical surface of human skin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.17.603990v1?rss=1">
<title>
<![CDATA[
Oxygen is Toxic in the Cold in C. elegans. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.17.603990v1?rss=1</link>
<description><![CDATA[
Temperature and oxygen are two factors that profoundly affect survival limits of animals; too much or too little of either is lethal. However, both humans and animals can exhibit exceptional survival when oxygen and temperature are simultaneously low. To better understand this apparent synergistic interaction between oxygen and temperature, we assayed the survival of Caenorhabditis elegans in experimental environments at different temperatures and oxygen concentrations. While nematodes cannot survive a day in room air at 2 {degrees}C (cold shock), we found that when oxygen is low, 200-fold less than room air, they can survive this temperature for 48 hours. Consistent with this, we found that worms exposed to high oxygen concentrations, 35 times greater than room air, are more sensitive to low temperature than worms in room air. These results show that normal atmospheric levels of oxygen are toxic in the cold. Using these survival assays, we found that cold acclimatization protects worms from lethal effects of high oxygen, mutations in the cold acclimatization pathway affect oxygen tolerance, and that the naturally occurring and physiologically relevant compounds glucose, manganese (II), and ascorbate improve survival limits in both low temperature and high oxygen when supplied early in life. These results show that the interdependence of temperature and oxygen on survival in C. elegans is based in part on shared physiological mechanisms involved in response to these two environmental stressors. The evolution and natural history of stress responses in animals suggest similar phenomena may function in humans.
]]></description>
<dc:creator>Suraci, C. M.</dc:creator>
<dc:creator>Morrison, M. L.</dc:creator>
<dc:creator>Roth, M. B.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.17.603990</dc:identifier>
<dc:title><![CDATA[Oxygen is Toxic in the Cold in C. elegans.]]></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/10.1101/2024.07.22.604631v1?rss=1">
<title>
<![CDATA[
Genetic variants affect diurnal glucose levels throughout the day 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.22.604631v1?rss=1</link>
<description><![CDATA[
Circadian rhythms not only coordinate the timing of wake and sleep but also regulate homeostasis within the body, including glucose metabolism. However, the genetic variants that contribute to temporal control of glucose levels have not been previously examined. Using data from 420,000 individuals from the UK Biobank and replicating our findings in 100,000 individuals from the Estonian Biobank, we show that diurnal serum glucose is under genetic control. We discover a robust temporal association of glucose levels at the Melatonin receptor 1B (MTNR1B) (rs10830963, P = 1e-22) and a canonical circadian pacemaker gene Cryptochrome 2 (CRY2) loci (rs12419690, P = 1e-16). Furthermore, we show that sleep modulates serum glucose levels and the genetic variants have a separate mechanism of diurnal control. Finally, we show that these variants independently modulate risk of type 2 diabetes. Our findings, together with earlier genetic and epidemiological evidence, show a clear connection between sleep and metabolism and highlight variation at MTNR1B and CRY2 as temporal regulators for glucose levels.
]]></description>
<dc:creator>Sinnott-Armstrong, N.</dc:creator>
<dc:creator>Strausz, S.</dc:creator>
<dc:creator>Urpa, L.</dc:creator>
<dc:creator>Abner, E.</dc:creator>
<dc:creator>Valliere, J.</dc:creator>
<dc:creator>Estonian Biobank Research Team,</dc:creator>
<dc:creator>Palta, P.</dc:creator>
<dc:creator>Dashti, H. S.</dc:creator>
<dc:creator>Daly, M.</dc:creator>
<dc:creator>Pritchard, J. K.</dc:creator>
<dc:creator>Saxena, R.</dc:creator>
<dc:creator>Jones, S. E.</dc:creator>
<dc:creator>Ollila, H. M.</dc:creator>
<dc:date>2024-07-23</dc:date>
<dc:identifier>doi:10.1101/2024.07.22.604631</dc:identifier>
<dc:title><![CDATA[Genetic variants affect diurnal glucose levels throughout the day]]></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/10.1101/2024.07.23.604849v1?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/10.1101/2024.07.23.604850v1?rss=1">
<title>
<![CDATA[
The battle of the sexes in humans is highly polygenic 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.23.604850v1?rss=1</link>
<description><![CDATA[
Sex-differential selection (SDS), which occurs when the fitness effects of alleles differ between males and females, can have profound impacts on the maintenance of genetic variation, disease risk, and other key aspects of natural populations. Because the sexes mix their autosomal genomes each generation, quantifying SDS is not possible using conventional population genetic approaches. Here, we introduce a novel method that exploits subtle sex differences in haplotype frequencies resulting from SDS acting in the current generation. Using data from 300K individuals in the UK Biobank, we estimate the strength of SDS throughout the genome. While only a handful of loci under SDS are individually significant, we uncover polygenic signals of genome-wide SDS for both viability and fecundity. An interesting life-history tradeoff emerges: alleles that increase viability more in one sex increase fecundity more in the other sex. Lastly, we find evidence of SDS on fecundity acting on alleles affecting arm fat-free mass. Taken together, our findings connect the long-standing evidence of SDS acting on human phenotypes with its impact on the genome.

Significance statementSelection often acts differently on females and males, as evidenced by the striking sexual dimorphism found in many taxa. As a result, alleles can have different fitness effects in each sex. Consequences can include higher levels of genetic variation and higher disease burdens in populations. This study introduces a novel method to quantify this sex-differential selection (SDS) and reveals that it acts throughout the human genome. We discovered a life history tradeoff between survival and fecundity in females and males and that SDS on fecundity acts on alleles affecting arm fat-free mass.
]]></description>
<dc:creator>Cole, J. M.</dc:creator>
<dc:creator>Scott, C. B.</dc:creator>
<dc:creator>Johnson, M. M.</dc:creator>
<dc:creator>Golightly, P. R.</dc:creator>
<dc:creator>Carlson, J.</dc:creator>
<dc:creator>Ming, M. J.</dc:creator>
<dc:creator>Harpak, A.</dc:creator>
<dc:creator>Kirkpatrick, M.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.23.604850</dc:identifier>
<dc:title><![CDATA[The battle of the sexes in humans is highly polygenic]]></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/10.1101/2024.07.24.604937v1?rss=1">
<title>
<![CDATA[
Stable centromere association of the yeast histone variant Cse4 requires its essential N-terminal domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.24.604937v1?rss=1</link>
<description><![CDATA[
Chromosome segregation relies on kinetochores that assemble on specialized centromeric chromatin containing a histone H3 variant. In budding yeast, a single centromeric nucleosome containing Cse4 assembles at a sequence-defined 125 bp centromere. Yeast centromeric sequences are poor templates for nucleosome formation in vitro, suggesting the existence of mechanisms that specifically stabilize Cse4 nucleosomes in vivo. The extended Cse4 N-terminal tail binds to the chaperone Scm3, and a short essential region called END within the N-terminal tail binds the inner kinetochore complex OA. To address the roles of these interactions, we utilized single molecule fluorescence assays to monitor Cse4 during kinetochore assembly. We found that OA and Scm3 independently stabilize Cse4 at centromeres via their END interaction. Scm3 binding to the Cse4 END is enhanced by Ipl1/Aurora B phosphorylation, identifying a previously unknown role for Ipl1 in ensuring Cse4 stability. Strikingly, an Ipl1 phosphomimetic mutation in the Cse4 END enhances Scm3 binding and can restore Cse4 recruitment in mutants defective in OA binding. Together, these data suggest that a key function of the essential Cse4 N-terminus is to ensure Cse4 localization at centromeres.
]]></description>
<dc:creator>Popchock, A. R.</dc:creator>
<dc:creator>Hedouin, S.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Asbury, C. L.</dc:creator>
<dc:creator>Stergachis, A. B.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.24.604937</dc:identifier>
<dc:title><![CDATA[Stable centromere association of the yeast histone variant Cse4 requires its essential N-terminal domain]]></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/10.1101/2024.07.25.605204v1?rss=1">
<title>
<![CDATA[
Decoding RNA Metabolism by RNA-linked CRISPR Screening in Human Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.25.605204v1?rss=1</link>
<description><![CDATA[
RNAs undergo a complex choreography of metabolic processes in human cells that are regulated by thousands of RNA-associated proteins. While the effects of individual RNA-associated proteins on RNA metabolism have been extensively characterized, the full complement of regulators for most RNA metabolic events remain unknown. Here we present a massively parallel RNA-linked CRISPR (ReLiC) screening approach to measure the responses of diverse RNA metabolic events to knockout of 2,092 human genes encoding all known RNA-associated proteins. ReLiC screens highlight modular interactions between gene networks regulating splicing, translation, and decay of mRNAs. When combined with biochemical fractionation of polysomes, ReLiC reveals striking pathway-specific coupling between growth fitness and mRNA translation. Perturbing different components of the translation and proteostasis machineries have distinct effects on ribosome occupancy, while perturbing mRNA transcription leaves ribosome occupancy largely intact. Isoform-selective ReLiC screens capture differential regulation of intron retention and exon skipping by SF3b complex subunits. Chemogenomic screens using ReLiC decipher translational regulators upstream of mRNA decay and uncover a role for the ribosome collision sensor GCN1 during treatment with the anti-leukemic drug homoharringtonine. Our work demonstrates ReLiC as a versatile platform for discovering and dissecting regulatory principles of human RNA metabolism.
]]></description>
<dc:creator>Nugent, P. J.</dc:creator>
<dc:creator>Park, H.</dc:creator>
<dc:creator>Wladyka, C. L.</dc:creator>
<dc:creator>Chen, K. Y.</dc:creator>
<dc:creator>Bynum, C.</dc:creator>
<dc:creator>Quarterman, G.</dc:creator>
<dc:creator>Hsieh, A. C.</dc:creator>
<dc:creator>Subramaniam, A. R.</dc:creator>
<dc:date>2024-07-26</dc:date>
<dc:identifier>doi:10.1101/2024.07.25.605204</dc:identifier>
<dc:title><![CDATA[Decoding RNA Metabolism by RNA-linked CRISPR Screening in Human Cells]]></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/10.1101/2024.08.03.606477v1?rss=1">
<title>
<![CDATA[
A targeted CRISPR screen identifies ETS1 as a regulator of HIV latency. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.03.606477v1?rss=1</link>
<description><![CDATA[
Human Immunodeficiency virus (HIV) infection is regulated by a wide array of host cell factors that combine to influence viral transcription and latency. To understand the complex relationship between the host cell and HIV latency, we performed a lentiviral CRISPR screen that targeted a set of host cell genes whose expression or activity correlates with HIV expression. We further investigated one of the identified factors - the transcription factor ETS1 and found that it is required for maintenance of HIV latency in a primary CD4 T cell model. Interestingly, ETS1 played divergent roles in actively infected and latently infected CD4 T cells, with knockout of ETS1 leading to reduced HIV expression in actively infected cells, but increased HIV expression in latently infected cells, indicating that ETS1 can play both a positive and negative role in HIV expression. CRISPR/Cas9 knockout of ETS1 in CD4 T cells from ART-suppressed people with HIV (PWH) confirmed that ETS1 maintains transcriptional repression of the clinical HIV reservoir. Transcriptomic profiling of ETS1-depleted cells from PWH identified a set of host cell pathways involved in viral transcription that are controlled by ETS1 in resting CD4 T cells. In particular, we observed that ETS1 knockout increased expression of the long non-coding RNA MALAT1 that has been previously identified as a positive regulator of HIV expression. Furthermore, the impact of ETS1 depletion on HIV expression in latently infected cells was partially dependent on MALAT1. Overall, these data demonstrate that ETS1 is an important regulator of HIV latency and influences expression of several cellular genes, including MALAT1, that could have a direct or indirect impact on HIV expression.

Author SummaryHIV latency is a major obstacle for the eradication of HIV. However, molecular mechanisms that restrict proviral expression during therapy are not well understood. Identification of host cell factors that silence HIV would create opportunities for targeting these factors to reverse latency and eliminate infected cells. Our study aimed to explore mechanisms of latency in infected cells by employing a lentiviral CRISPR screen and CRISPR/Cas9 knockout in primary CD4 T cells. These experiments revealed that ETS1 is essential for maintaining HIV latency in primary CD4 T cells and we further confirmed ETS1s role in maintaining HIV latency through CRISPR/Cas9 knockout in CD4 T cells from antiretroviral therapy (ART)-suppressed individuals with HIV. Transcriptomic profiling of ETS1-depleted cells from these individuals identified several host cell pathways involved in viral transcription regulated by ETS1, including the long non-coding RNA MALAT1. Overall, our study demonstrates that ETS1 is a critical regulator of HIV latency, affecting the expression of several cellular genes that directly or indirectly influence HIV expression.
]]></description>
<dc:creator>Ashokkumar, M.</dc:creator>
<dc:creator>Hafer, T. L.</dc:creator>
<dc:creator>Felton, A.</dc:creator>
<dc:creator>Archin, N.</dc:creator>
<dc:creator>Margolis, D. M.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Browne, E. P.</dc:creator>
<dc:date>2024-08-03</dc:date>
<dc:identifier>doi:10.1101/2024.08.03.606477</dc:identifier>
<dc:title><![CDATA[A targeted CRISPR screen identifies ETS1 as a regulator of HIV latency.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.21.608981v1?rss=1">
<title>
<![CDATA[
CARD8 inflammasome activation during HIV-1 cell-to-cell transmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.21.608981v1?rss=1</link>
<description><![CDATA[
Our previous work demonstrated that CARD8 detects HIV-1 infection by sensing the enzymatic activity of the HIV protease, resulting in CARD8-dependent inflammasome activation (Kulsuptrakul et al., 2023). CARD8 harbors a motif in its N-terminus that functions as a HIV protease substrate mimic, permitting innate immune recognition of HIV-1 protease activity, which when cleaved by HIV protease triggers CARD8 inflammasome activation. Here, we sought to understand CARD8 responses in the context of HIV-1 cell-to-cell transmission via a viral synapse. We observed that cell-to-cell transmission of HIV-1 between infected T cells and primary human monocyte-derived macrophages induces CARD8 inflammasome activation in a manner that is dependent on viral protease activity and largely independent of the NLRP3 inflammasome. Additionally, to further evaluate the viral determinants of CARD8 sensing, we tested a panel of HIV protease inhibitor resistant clones to establish how variation in HIV protease affects CARD8 activation. We identified mutant HIV-1 proteases that differentially cleave and activate CARD8 compared to wildtype HIV-1, thus indicating that natural variation in HIV protease affects not only the cleavage of the viral Gag-Pol polyprotein but also likely impacts innate sensing and inflammation.
]]></description>
<dc:creator>Kulsuptrakul, J.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Mitchell, P. S.</dc:creator>
<dc:date>2024-08-22</dc:date>
<dc:identifier>doi:10.1101/2024.08.21.608981</dc:identifier>
<dc:title><![CDATA[CARD8 inflammasome activation during HIV-1 cell-to-cell transmission]]></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/10.1101/2024.08.21.609082v1?rss=1">
<title>
<![CDATA[
A conserved opal termination codon optimizes a temperature-dependent tradeoff between protein production and processing in alphaviruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.21.609082v1?rss=1</link>
<description><![CDATA[
Alphaviruses are enveloped, single-stranded, positive-sense RNA viruses that often require transmission between arthropod and vertebrate hosts for their sustained propagation. Most alphaviruses encode an opal (UGA) termination codon in nonstructural protein 3 (nsP3) upstream of the viral polymerase, nsP4. The selective constraints underlying the conservation of the opal codon are poorly understood. Using primate and mosquito cells, we explored the role and selective pressure on the nsP3 opal codon through extensive mutational analysis in the prototype alphavirus, Sindbis virus (SINV). We found that the opal codon is highly favored over all other codons in primate cells under native 37{o}C growth conditions. However, this preference is diminished in mosquito and primate cells grown at a lower temperature. Thus, the primary determinant driving the selection of the opal stop codon is not host genetics but the passaging temperature. We show that the opal codon is preferred over amber and ochre termination codons because it results in the highest translational readthrough and polymerase production. However, substituting the opal codon with sense codons leads to excessive full-length polyprotein (P1234) production, which disrupts optimal nsP polyprotein processing, delays the switch from minus-strand to positive-strand RNA production, and significantly reduces SINV fitness at 37{degrees}C; this fitness defect is relieved at lower temperatures. A naturally occurring suppressor mutation unexpectedly compensates for a delayed transition from minus to genomic RNA production by also delaying the subsequent transition between genomic and sub-genomic RNA production. Our study reveals that the opal stop codon is the best solution for alphavirus replication at 37{o}C, producing enough nsP4 protein to maximize replication without disrupting nsP processing and RNA replication transitions needed for optimal fitness. Our study uncovers the intricate strategy dual-host alphaviruses use at a single codon to optimize fitness.
]]></description>
<dc:creator>Bhattacharya, T.</dc:creator>
<dc:creator>Alleman, E. M.</dc:creator>
<dc:creator>Noyola, A. C.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2024-08-22</dc:date>
<dc:identifier>doi:10.1101/2024.08.21.609082</dc:identifier>
<dc:title><![CDATA[A conserved opal termination codon optimizes a temperature-dependent tradeoff between protein production and processing in alphaviruses]]></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/10.1101/2024.08.21.609089v1?rss=1">
<title>
<![CDATA[
Post-translational modifications on protein VII are important during the early stages of adenovirus infection. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.21.609089v1?rss=1</link>
<description><![CDATA[
Due to the importance of post-translational modification (PTM) in cellular function, viruses have evolved to both take advantage of and be susceptible to such modification. Adenovirus encodes a multifunctional protein called protein VII, which is packaged with the viral genome in the core of virions and disrupts host chromatin during infection. Protein VII has several PTMs whose addition contributes to the subnuclear localization of protein VII. Here, we used mutant viruses that abrogate or mimic these PTMs on protein VII to interrogate their impact on protein VII function during adenovirus infection. We discovered that acetylation of the lysine in positions 2 or 3 (K2 or K3) is deleterious during early infection as mutation to alanine led to greater intake of protein VII to the nucleus and enhanced early gene expression. Furthermore, we determined that protein VII is acetylated at alternative residues late during infection which may compensate for the mutated sites. Lastly, due to the role of the early viral protein E1A in viral gene activation, we investigated the interaction between protein VII and E1A and demonstrated that protein VII interacts with E1A through a chromatin-mediated interaction. Together, these results emphasize that the complexity of virus-host interactions is intimately tied to post-translational modification.

ImportanceAdenoviruses are a ubiquitous human pathogen that cause a variety of diseases, such as respiratory infections, gastroenteritis, and conjunctivitis. While often viewed as a self-limiting infection in healthy individuals, adenoviruses are particularly harmful for immunocompromised patients. Here, we investigate the functional role of post-translational modifications (PTMs) on an essential adenovirus core protein, protein VII, describing how they regulate its function during early and late stages of infection. Our study focuses on how specific PTMs on protein VII influence transcription, localization, and interactions with other proteins, highlighting how PTMs are employed by viruses to alter protein function.
]]></description>
<dc:creator>Arnold, E. A.</dc:creator>
<dc:creator>Smith, J. R.</dc:creator>
<dc:creator>Leung, K.</dc:creator>
<dc:creator>Nguyen, D. H.</dc:creator>
<dc:creator>Kelnhofer-Millevolte, L. E.</dc:creator>
<dc:creator>Guo, M. S.</dc:creator>
<dc:creator>Smith, J. G.</dc:creator>
<dc:creator>Avgousti, D. C.</dc:creator>
<dc:date>2024-08-22</dc:date>
<dc:identifier>doi:10.1101/2024.08.21.609089</dc:identifier>
<dc:title><![CDATA[Post-translational modifications on protein VII are important during the early stages of adenovirus infection.]]></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/10.1101/2024.08.22.609143v1?rss=1">
<title>
<![CDATA[
Multiplexed Glycan Immunofluorescence Identification of Pancreatic Cancer Cell Subpopulations in Both Tumor and Blood Samples 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.22.609143v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDAC) tumor heterogeneity impedes the development of biomarker assays suitable for early disease detection that would improve patient outcomes. The CA19-9 glycan is currently used as a standalone biomarker for PDAC. Furthermore, previous studies have shown that cancer cells may display aberrant membrane-associated glycans. We therefore hypothesized that PDAC cancer cell subpopulations could be distinguished by aberrant glycan signatures. We used multiplexed glycan immunofluorescence combined with pathologist annotation and automated image processing to distinguish between PDAC cancer cell subpopulations within tumor tissue. Using a training-set/test-set approach, we found that PDAC cancer cells may be identified by signatures comprising 4 aberrant glycans (VVL, CA19-9, sTRA, and GM2) and that there are three glycan-defined PDAC tumor types: sTRA type, CA19-9 type, and intermixed. To determine whether the aberrant glycan signatures could be detected in blood samples, we developed hybrid glycan sandwich assays for membrane-associated glycans. In both patient-matched tumor and blood samples, the proportion of aberrant glycans detected was consistent. Furthermore, our multiplexed glycan immunofluorescent approach proved to be more sensitive and more specific than CA19-9 alone. Our results provide proof of concept for a novel methodology to improve early PDAC detection and patient outcomes.
]]></description>
<dc:creator>Binkowski, B.</dc:creator>
<dc:creator>Klamer, Z.</dc:creator>
<dc:creator>Gao, C.</dc:creator>
<dc:creator>Staal, B.</dc:creator>
<dc:creator>Repesh, A.</dc:creator>
<dc:creator>Tran, H.-L.</dc:creator>
<dc:creator>Brass, D. M.</dc:creator>
<dc:creator>Bartlett, P.</dc:creator>
<dc:creator>Gallinger, S.</dc:creator>
<dc:creator>Blomqvist, M.</dc:creator>
<dc:creator>Morrow, J. B.</dc:creator>
<dc:creator>Allen, P.</dc:creator>
<dc:creator>Shi, C.</dc:creator>
<dc:creator>Singhi, A.</dc:creator>
<dc:creator>Brand, R.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Hostetter, G.</dc:creator>
<dc:creator>Haab, B. B.</dc:creator>
<dc:date>2024-08-23</dc:date>
<dc:identifier>doi:10.1101/2024.08.22.609143</dc:identifier>
<dc:title><![CDATA[Multiplexed Glycan Immunofluorescence Identification of Pancreatic Cancer Cell Subpopulations in Both Tumor and Blood Samples]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.22.609229v1?rss=1">
<title>
<![CDATA[
Microenvironment T-Type calcium channels regulate neuronal and glial processes to promote glioblastoma growth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.22.609229v1?rss=1</link>
<description><![CDATA[
BackgroundGlioblastoma (GBM) is the most common primary malignant brain tumor. The aim of this study was to elucidate the role of microenvironment and intrinsic T-type calcium channels (Cav3) in regulating tumor growth and progression.

MethodsWe grafted syngeneic GBM cells into Cav3.2 knockout mice to assess the role of microenvironment T-Type calcium channels on GBM tumor growth. We performed single-cell RNA-seq (scRNA-seq) of tumors from WT and Cav3.2 KO mice to elucidate the regulation of tumors by the microenvironment. We used neurons from WT and Cav3.2 KO mice in co-culture with GBM stem cells (GSC) to assess the effects of Cav3.2 on neuron/GSC synaptic connections and tumor cell growth.

ResultsCav3.2 KO in the microenvironment led to significant reduction of GBM growth and prolongation of animal survival. scRNA-seq showed that microenvironment Cav3.2 regulates neuronal and glial biological processes. Microenvironment Cav3.2 downregulated numerous genes associated with regulating the OPC cell state in GBM tumors such as SOX10 and Olig2. Neuronal Cav3.2 promoted neuron/GSC synaptic connections and GSC growth. Treatment of GSCs with the Cav3 blocker mibefradil downregulated genes associated with neuronal processes. The Cav3 blocker drug mibefradil synergized with temozolomide (TMZ) and radiation to reduce in vivo tumor growth and prolong animal survival.

ConclusionsTogether these data reveal a role for microenvironment Cav3 in promoting GBM tumor progression through regulating neuronal and glial processes particularly associated with the OPC-cell state. Targeting both intrinsic and microenvironment Cav3 with the inhibitor mibefradil significantly enhanced the anti-GBM effects of TMZ and radiation.

Key PointsO_LIMicroenvironment Cav3.2 promotes GBM progression
C_LIO_LIMicroenvironment Cav3.2 promotes neuronal and glial processes
C_LIO_LIPharmacological targeting of intrinsic and microenvironment Cav3 synergizes with TMZ/radiation
C_LI

Importance of the StudyIn this study, we demonstrate for the first time that microenvironment Cav3.2 contributes to GBM progression and growth by regulating neuronal and glial processes. Our findings highlight the importance of T-type calcium channels in the microenvironment as well as the tumor and provides preclinically relevant data for the use of mibefradil to inhibit GBM growth in combination with standard of care therapies.
]]></description>
<dc:creator>Dube, C. J.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Saha, S.</dc:creator>
<dc:creator>Lai, M.</dc:creator>
<dc:creator>Gibert, M. K.</dc:creator>
<dc:creator>Escalante, M.</dc:creator>
<dc:creator>Hudson, K.</dc:creator>
<dc:creator>Wong, D.</dc:creator>
<dc:creator>Marcinkiewicz, P.</dc:creator>
<dc:creator>Yener, U.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Xu, E.</dc:creator>
<dc:creator>Sorot, A.</dc:creator>
<dc:creator>Mulcahy, E.</dc:creator>
<dc:creator>Kefas, B. A.</dc:creator>
<dc:creator>Hanif, F.</dc:creator>
<dc:creator>Guessous, F.</dc:creator>
<dc:creator>Patel, M. K.</dc:creator>
<dc:creator>Schiff, D.</dc:creator>
<dc:creator>Zong, H.</dc:creator>
<dc:creator>Purow, B. W.</dc:creator>
<dc:creator>Holland, E.</dc:creator>
<dc:creator>Sonkusare, S. K.</dc:creator>
<dc:creator>Sontheimer, H.</dc:creator>
<dc:creator>Abounader, R.</dc:creator>
<dc:date>2024-08-23</dc:date>
<dc:identifier>doi:10.1101/2024.08.22.609229</dc:identifier>
<dc:title><![CDATA[Microenvironment T-Type calcium channels regulate neuronal and glial processes to promote glioblastoma growth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.23.609293v1?rss=1">
<title>
<![CDATA[
Breastmilk IgG engages the neonatal immune system to instruct host-microbiota mutualism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.23.609293v1?rss=1</link>
<description><![CDATA[
Maternal antibodies fundamentally regulate infant immunity to the developing gut microbiota, yet the mechanisms underlying this process remain elusive. Here, we show that maternal IgG, ingested in the first week of life, functions to restrain microbiota-dependent adaptive immune responses and reduce offspring susceptibility to intestinal inflammation weeks later, following weaning. To exert these functions, efficient binding of IgG to gut bacterial antigens and engagement of Fc and complement dependent effector functions in offspring was required. These discoveries reveal a novel mechanism wherein maternal IgG engages the offspring immune system to calibrate responses to gut microbes. This mode of maternal immune instruction may provide adaptability to developmental shifts in microbiota necessary for establishing host-microbiota mutualism and limiting susceptibility to inflammatory disease.

One sentence abstractIngestion of maternal IgG during a discrete postnatal window calibrates neonatal immunity to the gut microbiota.
]]></description>
<dc:creator>Shenoy, M. K.</dc:creator>
<dc:creator>Rico, D.</dc:creator>
<dc:creator>Gordon, S.</dc:creator>
<dc:creator>Milburn, L.</dc:creator>
<dc:creator>Schwensen, J.</dc:creator>
<dc:creator>Caban, M.</dc:creator>
<dc:creator>Koch, M. A.</dc:creator>
<dc:date>2024-08-23</dc:date>
<dc:identifier>doi:10.1101/2024.08.23.609293</dc:identifier>
<dc:title><![CDATA[Breastmilk IgG engages the neonatal immune system to instruct host-microbiota mutualism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.23.609309v1?rss=1">
<title>
<![CDATA[
Chronic innate immune impairment and ZIKV persistence in the gastrointestinal tract during SIV infection in pigtail macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.23.609309v1?rss=1</link>
<description><![CDATA[
Mosquito borne flaviviruses, including dengue (DENV) and Zika (ZIKV) viruses, have caused global epidemics in areas with high HIV prevalence due to the expanded geographic range of arthropod vectors. Despite the occurrence of large flavivirus outbreaks in countries with high HIV prevalence, there is little knowledge regarding the effects of flavivirus infection in people living with HIV (PLWH). Here, we use a pigtail macaque model of HIV/AIDS to investigate the impact of simian immunodeficiency virus (SIV)-induced immunosuppression on ZIKV replication and pathogenesis. Early acute SIV infection induced expansion of peripheral ZIKV cellular targets and increased innate immune activation and peripheral blood mononuclear cells (PBMC) from SIV infected macaques were less permissive to ZIKV infection in vitro. In SIV-ZIKV co-infected animals, we found increased persistence of ZIKV in the periphery and tissues corresponding to alterations in innate cellular (monocytes, neutrophils) recruitment to the blood and tissues, decreased anti-ZIKV immunity, and chronic peripheral inflammatory and innate immune gene expression. Collectively, these findings suggest that untreated SIV infection may impair cellular innate responses and create an environment of chronic immune activation that promotes prolonged ZIKV viremia and persistence in the gastrointestinal tract. These results suggest that PLWH or other immunocompromised individuals could be at a higher risk for chronic ZIKV replication, which in turn could increase the timeframe of ZIKV transmission. Thus, PLWH are important populations to target during the deployment of vaccine and treatment strategies against ZIKV.

Author SummaryFlaviviruses, including Zika virus (ZIKV), cause global epidemics in areas with high HIV prevalence. Yet questions remain as to whether ZIKV disease is altered during an immunocompromised state and the potential immune mechanisms contributing to enhanced disease. This is essential to our understanding of ZIKV disease in people living with HIV (PLWH). Here, we use a non-human primate (NHP) model of HIV/AIDS to investigate the impact of immune suppression on ZIKV replication and pathogenesis. The use of the NHP model was critical for the assessment of longitudinal specimens across tissues that are active sites of flavivirus replication and host immune responses. This study broadly demonstrates that ZIKV pathogenesis is altered and more persistent in states of immunosuppression. Collectively, this study suggests that in PLWH and immunocompromised individuals, other arboviruses, including dengue and West Nile viruses, could similarly alter pathogenesis and/or viral peristance in tissues. Furthermore, this study highlights the need to prioritize immunocompromised individuals in the design and rollout of vaccines against arboviral diseases.
]]></description>
<dc:creator>Tisoncik-Go, J.</dc:creator>
<dc:creator>Lewis, T. B.</dc:creator>
<dc:creator>Whitmore, L. S.</dc:creator>
<dc:creator>Voss, K.</dc:creator>
<dc:creator>Niemeyer, S.</dc:creator>
<dc:creator>Dai, J.</dc:creator>
<dc:creator>Kim, P.</dc:creator>
<dc:creator>Hubbell, K.</dc:creator>
<dc:creator>Iwayama, N.</dc:creator>
<dc:creator>Ahrens, C.</dc:creator>
<dc:creator>Wangari, S.</dc:creator>
<dc:creator>Murnane, R.</dc:creator>
<dc:creator>Edlefsen, P. T.</dc:creator>
<dc:creator>Guerriero, K. A.</dc:creator>
<dc:creator>Gale, M.</dc:creator>
<dc:creator>Fuller, D. H.</dc:creator>
<dc:creator>O'Connor, M.</dc:creator>
<dc:date>2024-08-24</dc:date>
<dc:identifier>doi:10.1101/2024.08.23.609309</dc:identifier>
<dc:title><![CDATA[Chronic innate immune impairment and ZIKV persistence in the gastrointestinal tract during SIV infection in pigtail macaques]]></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/10.1101/2024.08.25.607996v1?rss=1">
<title>
<![CDATA[
Molecular basis of pathogenicity of the recently emerged FCoV-23 coronavirus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.25.607996v1?rss=1</link>
<description><![CDATA[
The ability of coronaviruses to recombine and cross species barriers affects human and animal health globally and is a pandemic threat. FCoV-23 is a recently emerged, highly pathogenic recombinant coronavirus responsible for a widespread outbreak of feline infectious peritonitis (FIP) likely linked to in-host viral evolution. Here, we report cryoEM structures of two FCoV-23 spike (S) isoforms explaining that the in-host loss of domain 0 observed in clinical samples enhances entry into cells and fusogenicity by facilitating protease access, leading to biotype switching and lethality. We show that FCoV-23 can use several aminopeptidase N (APN) orthologs as receptors and reveal the molecular determinants of receptor species tropism, including a glycan modulating human receptor utilization. We define antigenic relationships among alphacoronaviruses infecting humans and other mammalian species and identify a cross-reactive alphacoronavirus monoclonal antibody inhibiting FCoV-23 pseudovirus entry, paving the way for vaccine and therapeutic development targeting this highly pathogenic virus.
]]></description>
<dc:creator>Tortorici, M. A.</dc:creator>
<dc:creator>Choi, A.</dc:creator>
<dc:creator>Gibson, C.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Brown, J. T.</dc:creator>
<dc:creator>Stewart, C.</dc:creator>
<dc:creator>Joshi, A.</dc:creator>
<dc:creator>Harari, S.</dc:creator>
<dc:creator>Willoughby, I.</dc:creator>
<dc:creator>Treichel, C.</dc:creator>
<dc:creator>Leif, E.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:creator>Tait-Burkard, C.</dc:creator>
<dc:creator>Whittaker, G. R.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:date>2024-08-26</dc:date>
<dc:identifier>doi:10.1101/2024.08.25.607996</dc:identifier>
<dc:title><![CDATA[Molecular basis of pathogenicity of the recently emerged FCoV-23 coronavirus]]></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/10.1101/2024.08.29.610392v1?rss=1">
<title>
<![CDATA[
Spatiotemporal dynamics of tumor-CAR T-cell interaction following local administration in solid cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.29.610392v1?rss=1</link>
<description><![CDATA[
The success of chimeric antigen receptor (CAR) T-cell therapy in treating hematologic malignancies has generated widespread interest in translating this technology to solid cancers. However, issues like tumor infiltration, the immunosuppressive tumor microenvironment, and tumor heterogeneity limit its efficacy in the solid tumor setting. Recent experimental and clinical studies propose local administration directly into the tumor or at the tumor site to increase CAR T-cell infiltration and improve treatment outcomes. Characteristics of the types of solid tumors that may be the most receptive to this treatment approach remain unclear. In this work, we develop a spatiotemporal model for CAR T-cell treatment of solid tumors, and use numerical simulations to compare the effect of introducing CAR T cells via intratumoral injection versus intracavitary administration in diverse cancer types. We demonstrate that the model can recapitulate tumor and CAR T-cell data from imaging studies of local administration of CAR T cells in mouse models. Our results suggest that locally administered CAR T cells will be most successful against slowly proliferating, highly diffusive tumors, which have the lowest average tumor cell density. These findings affirm the clinical observation that CAR T cells will not perform equally across different types of solid tumors, and suggest that measuring tumor density may be helpful when considering the feasibility of CAR T-cell therapy and planning dosages for a particular patient. We additionally find that local delivery of CAR T cells can result in deep tumor responses, provided that the initial CAR T-cell dose does not contain a significant fraction of exhausted cells.
]]></description>
<dc:creator>Owens, K.</dc:creator>
<dc:creator>Rahman, A.</dc:creator>
<dc:creator>Bozic, I.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.29.610392</dc:identifier>
<dc:title><![CDATA[Spatiotemporal dynamics of tumor-CAR T-cell interaction following local administration in solid cancers]]></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/10.1101/2024.08.30.610517v1?rss=1">
<title>
<![CDATA[
Integrator complex subunit 12 knockout overcomes a transcriptional block to HIV latency reversal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.30.610517v1?rss=1</link>
<description><![CDATA[
The latent HIV reservoir is a major barrier to HIV cure. Combining latency reversal agents (LRAs) with differing mechanisms of action such as AZD5582, a non-canonical NF-kB activator, and I-BET151, a bromodomain inhibitor is appealing towards inducing HIV-1 reactivation. However, even this LRA combination needs improvement as it is inefficient at activating proviruses in cells from people living with HIV (PLWH). We performed a CRISPR screen in conjunction with AZD5582 & I-BET151 and identified a member of the Integrator complex as a target to improve this LRA combination, specifically Integrator complex subunit 12 (INTS12). Integrator functions as a genome-wide attenuator of transcription that acts on elongation through its RNA cleavage and phosphatase modules. Knockout of INTS12 improved latency reactivation at the transcriptional level and is more specific to the HIV-1 provirus than AZD5582 & I-BET151 treatment alone. We found that INTS12 is present on chromatin at the promoter of HIV and therefore its effect on HIV may be direct. Additionally, we observed more RNAPII in the gene body of HIV only with the combination of INTS12 knockout with AZD5582 & I-BET151, indicating that INTS12 induces a transcriptional elongation block to viral reactivation. Moreover, knockout of INTS12 increased HIV-1 reactivation in CD4 T cells from virally suppressed PLWH ex vivo, and we detected viral RNA in the supernatant from CD4 T cells of all three virally suppressed PLWH tested upon INTS12 knockout suggesting that INTS12 prevents full-length HIV RNA production in primary T cells. Finally, we found that INTS12 more generally limits the efficacy of a variety of LRAs with different mechanisms of action.
]]></description>
<dc:creator>Gray, C. N.</dc:creator>
<dc:creator>Ashokkumar, M.</dc:creator>
<dc:creator>Janssens, D. H.</dc:creator>
<dc:creator>Kirchherr, J. L.</dc:creator>
<dc:creator>Allard, B.</dc:creator>
<dc:creator>Hsieh, E.</dc:creator>
<dc:creator>Hafer, T. L.</dc:creator>
<dc:creator>Archin, N. M.</dc:creator>
<dc:creator>Browne, E. P.</dc:creator>
<dc:creator>Emerman, M.</dc:creator>
<dc:date>2024-08-31</dc:date>
<dc:identifier>doi:10.1101/2024.08.30.610517</dc:identifier>
<dc:title><![CDATA[Integrator complex subunit 12 knockout overcomes a transcriptional block to HIV latency reversal]]></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/10.1101/2024.09.04.611299v1?rss=1">
<title>
<![CDATA[
RCC1 depletion drives protein transport defects and rupture in micronuclei 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.04.611299v1?rss=1</link>
<description><![CDATA[
Micronuclei (MN) are a commonly used marker of chromosome instability that form when missegregated chromatin recruits its own nuclear envelope (NE) after mitosis. MN frequently rupture, which results in genome instability, upregulation of metastatic genes, and increased immune signaling. MN rupture is linked to NE defects, but the cause of these defects is poorly understood. Previous work from our lab found that chromosome identity correlates with rupture timing for small MN, i.e. MN containing a short chromosome, with more euchromatic chromosomes forming more stable MN with fewer nuclear lamina gaps. Here we demonstrate that histone methylation promotes rupture and nuclear lamina defects in small MN. This correlates with increased MN size, and we go on to find that all MN have a constitutive nuclear export defect that drives MN growth and nuclear lamina gap expansion, making the MN susceptible to rupture. We demonstrate that these export defects arise from decreased RCC1 levels in MN and that additional loss of RCC1 caused by low histone methylation in small euchromatic MN results in additional import defects that suppress nuclear lamina gaps and MN rupture. Through analysis of mutational signatures associated with early and late rupturing chromosomes in the Pan-Cancer Analysis of Whole Genomes (PCAWG) dataset, we identify an enrichment of APOBEC and DNA polymerase E hypermutation signatures in chromothripsis events on early and mid rupturing chromosomes, respectively, suggesting that MN rupture timing could determine the landscape of structural variation in chromothripsis. Our study defines a new model of MN rupture where increased MN growth, caused by defects in protein export, drives gaps in nuclear lamina organization that make the MN susceptible to membrane rupture with long-lasting effects on genome architecture.
]]></description>
<dc:creator>Zych, M. G.</dc:creator>
<dc:creator>Contreras, M.</dc:creator>
<dc:creator>Vashisth, M.</dc:creator>
<dc:creator>Mammel, A. E.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:creator>Hatch, E. M.</dc:creator>
<dc:date>2024-09-05</dc:date>
<dc:identifier>doi:10.1101/2024.09.04.611299</dc:identifier>
<dc:title><![CDATA[RCC1 depletion drives protein transport defects and rupture in micronuclei]]></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/10.1101/2024.09.04.611164v1?rss=1">
<title>
<![CDATA[
High Resolution Class I HLA -A, -B, and -C Diversity in Eastern and Southern African Populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.04.611164v1?rss=1</link>
<description><![CDATA[
Africa remains significantly underrepresented in high-resolution Human Leukocyte Antigen (HLA) data, despite being one of the most genetically diverse regions in the world. This critical gap in genetic information poses a substantial barrier to HLA-based research on the continent. In this study, Class I HLA data from Eastern and Southern African populations were analysed to assess genetic diversity across the region. We examined allele and haplotype frequency distributions, deviations from Hardy-Weinberg Equilibrium (HWE), linkage disequilibrium (LD), and conducted neutrality tests of homozygosity across various populations. Additionally, the African HLA data were compared to those of Caucasian and African American populations using the Jaccard index and multidimensional scaling (MDS) methods. The study revealed that South African populations exhibited 50.4% more genetic diversity within the Class I HLA region compared to other African populations. Zambia showed an estimated 36.5% genetic diversity, with Kenya, Rwanda and Uganda showing 35.7%, 34.2%, and 31.1%, respectively. Furthermore, an analysis of in-country diversity among diferent tribes indicated an average Class I HLA diversity of 25.7% in Kenya, 17% in Rwanda, 2.8% in South Africa, 13.6% in Uganda, and 6.5% in Zambia. The study also highlighted the genetic distinctness of Caucasian and African American populations compared to African populations. Notably, the diferential frequencies of disease- promoting and disease-preventing HLA alleles across these populations emphasize the urgent need to generate high-quality HLA data for all regions of Africa and its major ethnic groups. Such eforts will be crucial in enhancing healthcare outcomes across the continent.

Author SummaryThis study investigated the diversity of class I HLA in the eastern and southern regions of the African continent using a population genetics approach. Analysis of HLA data at both country and tribal levels revealed significant genetic diferences and the unique characteristics of these populations compared to Caucasian and African American populations in the United States. The diferential frequencies of disease-promoting and disease-preventing HLA alleles across these populations suggest that large-scale vaccine administration may be inefective without a thorough understanding of the HLA composition of each population. This study highlights the urgent need to generate high-quality HLA data across all regions of Africa and its major ethnic groups. Such comprehensive data collection is essential for optimizing vaccine design, deepening our understanding of HLA-disease associations, and ultimately improving healthcare outcomes across the continent.
]]></description>
<dc:creator>Ndhlovu, Z.</dc:creator>
<dc:creator>Banjoko, A. W.</dc:creator>
<dc:creator>Ng'uni, T.</dc:creator>
<dc:creator>Naidoo, N.</dc:creator>
<dc:creator>Ramsuran, V.</dc:creator>
<dc:creator>Hyrien, O.</dc:creator>
<dc:date>2024-09-08</dc:date>
<dc:identifier>doi:10.1101/2024.09.04.611164</dc:identifier>
<dc:title><![CDATA[High Resolution Class I HLA -A, -B, and -C Diversity in Eastern and Southern African Populations]]></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/10.1101/2024.09.04.611290v1?rss=1">
<title>
<![CDATA[
Putting computational models of immunity to the test - an invited challenge to predict B. pertussis vaccination outcomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.04.611290v1?rss=1</link>
<description><![CDATA[
Systems vaccinology studies have been used to build computational models that predict individual vaccine responses and identify the factors contributing to differences in outcome. Comparing such models is challenging due to variability in study designs. To address this, we established a community resource to compare models predicting B. pertussis booster responses and generate experimental data for the explicit purpose of model evaluation. We here describe our second computational prediction challenge using this resource, where we benchmarked 49 algorithms from 53 scientists. We found that the most successful models stood out in their handling of nonlinearities, reducing large feature sets to representative subsets, and advanced data preprocessing. In contrast, we found that models adopted from literature that were developed to predict vaccine antibody responses in other settings performed poorly, reinforcing the need for purpose-built models. Overall, this demonstrates the value of purpose-generated datasets for rigorous and open model evaluations to identify features that improve the reliability and applicability of computational models in vaccine response prediction.
]]></description>
<dc:creator>Shinde, P.</dc:creator>
<dc:creator>Willemsen, L.</dc:creator>
<dc:creator>Anderson, M.</dc:creator>
<dc:creator>Aoki, M.</dc:creator>
<dc:creator>Basu, S.</dc:creator>
<dc:creator>Burel, J. G.</dc:creator>
<dc:creator>Cheng, P.</dc:creator>
<dc:creator>Dastidar, S. G.</dc:creator>
<dc:creator>Dunleavy, A.</dc:creator>
<dc:creator>Einav, T.</dc:creator>
<dc:creator>Forschmiedt, J.</dc:creator>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Garcia, J.</dc:creator>
<dc:creator>Gibson, W.</dc:creator>
<dc:creator>Greenbaum, J. A.</dc:creator>
<dc:creator>Guan, L.</dc:creator>
<dc:creator>Guan, W.</dc:creator>
<dc:creator>Gygi, J. P.</dc:creator>
<dc:creator>Ha, B.</dc:creator>
<dc:creator>Hou, J.</dc:creator>
<dc:creator>Hsiao, J.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Jansen, R.</dc:creator>
<dc:creator>Kakoty, B.</dc:creator>
<dc:creator>Kang, Z.</dc:creator>
<dc:creator>Kobie, J. J.</dc:creator>
<dc:creator>Kojima, M.</dc:creator>
<dc:creator>Konstorum, A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Lewis, S. A.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Lock, E. F.</dc:creator>
<dc:creator>Mahita, J.</dc:creator>
<dc:creator>Mendes, M.</dc:creator>
<dc:creator>Meng, H.</dc:creator>
<dc:creator>Neher, A.</dc:creator>
<dc:creator>Nili, S.</dc:creator>
<dc:creator>Orfield, S.</dc:creator>
<dc:creator>Overton, J.</dc:creator>
<dc:creator>Pai, N.</dc:creator>
<dc:creator>Parker, C.</dc:creator>
<dc:creator>Qian, B.</dc:creator>
<dc:creator>Rasmussen, M.</dc:creator>
<dc:creator>Reyna, J.</dc:creator>
<dc:creator>Richardson, E.</dc:creator>
<dc:creator>Safo, S.</dc:creator>
<dc:creator>Sorenson, J.</dc:creator>
<dc:creator>Srinivasan, A.</dc:creator>
<dc:creator>Thrupp, N.</dc:creator>
<dc:creator>Tip</dc:creator>
<dc:date>2024-09-08</dc:date>
<dc:identifier>doi:10.1101/2024.09.04.611290</dc:identifier>
<dc:title><![CDATA[Putting computational models of immunity to the test - an invited challenge to predict B. pertussis vaccination outcomes]]></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/10.1101/2024.09.05.611179v1?rss=1">
<title>
<![CDATA[
Rare twin cysteine residues in the HIV-1 envelope variable region 1 link to neutralization escape and breadth development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.05.611179v1?rss=1</link>
<description><![CDATA[
The identification of HIV-1 Envelope glycoprotein (Env) traits associated with development of neutralization cross-reactivity in natural infection is critical for vaccine design. Here we describe the presence of additional Cysteine (Cys) residues in V1 that are enriched among people with elite neutralization breadth. Using >65,000 V1 sequences from the CATNAP database, the AMP trials and three large longitudinal HIV infection cohorts, the SHCS, ZPHI and CAPRISA studies, we show that Env variants with extra V1 Cys are present at low levels throughout infection and fluctuate in frequency over time within participants. We demonstrate an independent association of extra V1 Cys with elite plasma neutralization, and a strong preference for two versus one extra Cys, suggesting certain Envs introduce an additional disulfide bond for stabilization. We observed high levels of neutralization resistance among Envs from 34 bNAb donors, of which 17.6% had elongated V1 regions with extra Cys. We show that extra V1 Cys moderately increase neutralization resistance in an Env from a V2- Apex bNAb-inducer. Modulation of the accessibility of bNAb epitopes on this Env by extra V1 Cys enhanced epitope shielding of several regions, but increased V2 exposure. This suggests that escape from autologous neutralizing activity drove insertion of the extra V1 Cys, creating a modified antigen that may have favored V2 bNAb induction in this donor. Overall, we identify a rare motif of twin Cys in V1 that confers increased neutralization resistance and Env stabilization, is associated with bNAb induction, and may hold potential for incorporation into future HIV bNAb immunogens.
]]></description>
<dc:creator>Hesselman, M. C.</dc:creator>
<dc:creator>Zeeb, M.</dc:creator>
<dc:creator>Rusert, P.</dc:creator>
<dc:creator>Pasin, C.</dc:creator>
<dc:creator>Mamrosh, J.</dc:creator>
<dc:creator>Kariuki, S.</dc:creator>
<dc:creator>Sickmann, M.</dc:creator>
<dc:creator>Kaufmann, M. M.</dc:creator>
<dc:creator>Schmidt, D.</dc:creator>
<dc:creator>Friedrich, N.</dc:creator>
<dc:creator>Metzner, K. J.</dc:creator>
<dc:creator>Rindler, A.</dc:creator>
<dc:creator>Kuster, H.</dc:creator>
<dc:creator>Adams, C.</dc:creator>
<dc:creator>Thebus, R.</dc:creator>
<dc:creator>Huber, M.</dc:creator>
<dc:creator>Yerly, S.</dc:creator>
<dc:creator>Leuzinger, K.</dc:creator>
<dc:creator>Perreau, M.</dc:creator>
<dc:creator>Koller, R.</dc:creator>
<dc:creator>Dollenmaier, G.</dc:creator>
<dc:creator>Frigerio, S.</dc:creator>
<dc:creator>Westfall, D. H.</dc:creator>
<dc:creator>Deng, W.</dc:creator>
<dc:creator>DeCamp, A. C.</dc:creator>
<dc:creator>Juraska, M.</dc:creator>
<dc:creator>Edupuganti, S.</dc:creator>
<dc:creator>Mgodi, N.</dc:creator>
<dc:creator>Murrell, H.</dc:creator>
<dc:creator>Garrett, N.</dc:creator>
<dc:creator>Wagh, K.</dc:creator>
<dc:creator>Mullins, J. I.</dc:creator>
<dc:creator>Williamson, C.</dc:creator>
<dc:creator>Moore, P. L.</dc:creator>
<dc:creator>Günthard, H. F.</dc:creator>
<dc:creator>Kouyos, R. D.</dc:creator>
<dc:creator>Trkola, A.</dc:creator>
<dc:date>2024-09-10</dc:date>
<dc:identifier>doi:10.1101/2024.09.05.611179</dc:identifier>
<dc:title><![CDATA[Rare twin cysteine residues in the HIV-1 envelope variable region 1 link to neutralization escape and breadth development]]></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/10.1101/2024.09.09.611517v1?rss=1">
<title>
<![CDATA[
Merkel cell carcinoma-derived macrophage migration inhibitory factor (MIF) may promote persistence of Chronic Lymphocytic Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.09.611517v1?rss=1</link>
<description><![CDATA[
While concurrent diagnoses of Merkel cell carcinoma (MCC) and other cancers, like Chronic lymphocytic leukemia (CLL), are rare, patients with MCC have a 30-fold higher incidence of CLL. While these increases have been attributed to the ability of CLL to suppress immune responses allowing for the emergence of MCC, here we found evidence that MCC could support the persistence of CLL. Using single cell sequencing approaches and computational analyses of MCC and CLL from a patient where both cancers were present in the same lymph node, we found that production of macrophage migration inhibitory factor (MIF) by MCC could promote the persistence of CLL through stimulation of CD74 and CXCR4. These results may explain why blood cell counts rapidly normalized after treatment for MCC and were maintained at normal levels despite the absence of treatment for CLL.



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org.highwire.dtl.DTLVardef@188968forg.highwire.dtl.DTLVardef@54c084org.highwire.dtl.DTLVardef@2b3a14org.highwire.dtl.DTLVardef@1f20eba_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Alencar, G. F.</dc:creator>
<dc:creator>Rodriguez, H. J.</dc:creator>
<dc:creator>Pulliam, T. H.</dc:creator>
<dc:creator>Remington, A. S.</dc:creator>
<dc:creator>Gilmour, M. W.</dc:creator>
<dc:creator>Alam, R.</dc:creator>
<dc:creator>Jabbour, A. J.</dc:creator>
<dc:creator>Mullen, L. J.</dc:creator>
<dc:creator>DeBuysscher, B. L.</dc:creator>
<dc:creator>Nghiem, P.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:date>2024-09-13</dc:date>
<dc:identifier>doi:10.1101/2024.09.09.611517</dc:identifier>
<dc:title><![CDATA[Merkel cell carcinoma-derived macrophage migration inhibitory factor (MIF) may promote persistence of Chronic Lymphocytic Leukemia]]></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/10.1101/2024.09.17.613536v1?rss=1">
<title>
<![CDATA[
The Drosophila maternal-effect gene abnormal oocyte (ao) does not repress histone gene expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.17.613536v1?rss=1</link>
<description><![CDATA[
The abnormal oocyte (ao) gene of Drosophila melanogaster is a maternal-effect lethal gene previously identified as encoding a transcriptional regulator of core histones. However, background genetic mutations in existing ao mutant strains could compromise their utility in manipulating histone levels. To distinguish the true ao phenotype from background effects, we created two new ao reagents: a CRISPR/Cas9-mediated knockout of the ao allele for genetic and molecular analyses and an epitope-tagged ao allele for cytological experiments. Using these reagents, we confirm previous findings that loss of ao causes maternal-effect lethality, which can be rescued by either a decrease in the histone gene copy number or by Y chromosome heterochromatin. Our data indicate that ao genetically interacts with the heterochromatin, as previously suggested. However, contrary to a prior study, we find neither Ao localization to histone genes nor ao repression of core histone transcript levels. Thus, the molecular basis for ao-associated maternal-effect lethality remains unknown.

Article SummaryA series of foundational papers established that abnormal oocyte (ao), a euchromatic maternal-effect lethal gene, interacts with heterochromatin and the histone multigene cluster to dictate embryonic viability in D. melanogaster. An earlier report argued that ao encodes a protein that localizes to and represses histone gene expression, thereby connecting histone gene overexpression with ao mutant maternal-effect lethality. Using new reagents for genetics and cytology, we recapitulate findings that ao encodes a maternal-effect lethal gene, whose loss is ameliorated by excess heterochromatin or loss of histone genes. However, we find that ao does not affect histone gene expression. Thus, how ao loss causes maternal-effect lethality remains unknown.
]]></description>
<dc:creator>Takenaka, R.</dc:creator>
<dc:creator>Simmerman, S. M.</dc:creator>
<dc:creator>Schmidt, C. A.</dc:creator>
<dc:creator>Albanese, E. H.</dc:creator>
<dc:creator>Rieder, L. E.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2024-09-18</dc:date>
<dc:identifier>doi:10.1101/2024.09.17.613536</dc:identifier>
<dc:title><![CDATA[The Drosophila maternal-effect gene abnormal oocyte (ao) does not repress histone gene expression]]></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/10.1101/2024.09.17.613111v1?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/10.1101/2024.09.18.612965v1?rss=1">
<title>
<![CDATA[
Overcoming immune evasion from post-translational modification of a mutant KRAS epitope to achieve TCR-engineered T cell-mediated antitumor activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.18.612965v1?rss=1</link>
<description><![CDATA[
T cell receptor (TCR)-T cell immunotherapy, in which T cells are engineered to express a TCR targeting a tumor epitope, is a form of adoptive cell therapy (ACT) that has exhibited promise against various tumor types. Mutants of oncoprotein KRAS, particularly at glycine-12 (G12), are frequent drivers of tumorigenicity, making them attractive targets for TCR-T cell therapy. However, class I-restricted TCRs specifically targeting G12-mutant KRAS epitopes in the context of tumors expressing HLA-A2, the most common human HLA-A allele, have remained elusive despite evidence an epitope encompassing such mutations can bind HLA-A2 and induce T cell responses. We report post-translational modifications (PTMs) on this epitope may allow tumor cells to evade immunologic pressure from TCR-T cells. A lysine side chain-methylated KRASG12V peptide, rather than the unmodified epitope, may be presented in HLA-A2 by tumor cells and impact TCR recognition. Using a novel computationally guided approach, we developed by mutagenesis TCRs that recognize this methylated peptide, enhancing tumor recognition and destruction. Additionally, we identified TCRs with similar functional activity in normal repertoires from primary T cells by stimulation with modified peptide, clonal expansion, and selection. Mechanistically, a gene knockout screen to identify mechanism(s) by which tumor cells methylate/demethylate this epitope unveiled SPT6 as a demethylating protein that could be targeted to improve effectiveness of these new TCRs. Our findings highlight the role of PTMs in immune evasion and suggest identifying and targeting such modifications should make effective ACTs available for a substantially greater range of tumors than the current therapeutic landscape.

One-sentence summaryTumor cell methylation of KRASG12V epitope in HLA-A2 permits immune evasion, and new TCRs were generated to overcome this with engineered cell therapy.
]]></description>
<dc:creator>Lee, J. W.</dc:creator>
<dc:creator>Chen, E. Y.</dc:creator>
<dc:creator>Hu, T.</dc:creator>
<dc:creator>Perret, R.</dc:creator>
<dc:creator>Chaffee, M. E.</dc:creator>
<dc:creator>Martinov, T.</dc:creator>
<dc:creator>Mureli, S.</dc:creator>
<dc:creator>McCurdy, C. L.</dc:creator>
<dc:creator>Jones, L. A.</dc:creator>
<dc:creator>Gafken, P. R.</dc:creator>
<dc:creator>Chanana, P.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Chapuis, A. G.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Schmitt, T. M.</dc:creator>
<dc:creator>Greenberg, P. D.</dc:creator>
<dc:date>2024-09-22</dc:date>
<dc:identifier>doi:10.1101/2024.09.18.612965</dc:identifier>
<dc:title><![CDATA[Overcoming immune evasion from post-translational modification of a mutant KRAS epitope to achieve TCR-engineered T cell-mediated antitumor activity]]></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/10.1101/2024.09.21.614255v1?rss=1">
<title>
<![CDATA[
MAX inactivation deregulates the MYC network and induces neuroendocrine neoplasia in multiple tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.21.614255v1?rss=1</link>
<description><![CDATA[
The MYC transcription factor requires MAX for DNA binding and widespread activation of gene expression in both normal and neoplastic cells. Surprisingly, inactivating mutations in MAX are associated with a subset of neuroendocrine cancers including pheochromocytoma, pituitary adenoma and small cell lung cancer. Neither the extent nor the mechanisms of MAX tumor suppression are well understood. Deleting Max across multiple mouse neuroendocrine tissues, we find Max inactivation alone produces pituitary adenomas while Max loss cooperates with Rb1/Trp53 loss to accelerate medullary thyroid C-cell and pituitary adenoma development. In the thyroid tumor cell lines, MAX loss triggers a striking shift in genomic occupancy by other members of the MYC network (MNT, MLX, MondoA) supporting metabolism, survival and proliferation of neoplastic neuroendocrine cells. Our work reveals MAX as a broad suppressor of neuroendocrine tumorigenesis through its ability to maintain a balance of genomic occupancies among the diverse transcription factors in the MYC network.

TeaserMAX inactivation deregulates multiple transcription factors to induce neuroendocrine cancers
]]></description>
<dc:creator>Freie, B.</dc:creator>
<dc:creator>Ibrahim, A. H.</dc:creator>
<dc:creator>Carroll, P. A.</dc:creator>
<dc:creator>Bronson, R. T.</dc:creator>
<dc:creator>Augert, A.</dc:creator>
<dc:creator>MacPherson, D.</dc:creator>
<dc:creator>Eisenman, R. N.</dc:creator>
<dc:date>2024-09-24</dc:date>
<dc:identifier>doi:10.1101/2024.09.21.614255</dc:identifier>
<dc:title><![CDATA[MAX inactivation deregulates the MYC network and induces neuroendocrine neoplasia in multiple tissues]]></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/10.1101/2024.09.24.612755v1?rss=1">
<title>
<![CDATA[
Developmental regulation of endothelial-to-hematopoietic transition from induced pluripotent stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.24.612755v1?rss=1</link>
<description><![CDATA[
Hematopoietic stem cells (HSCs) arise in embryogenesis from a specialized hemogenic endothelium (HE). In this process, HE cells undergo a unique fate change termed endothelial-to-hematopoietic transition, or EHT. While induced pluripotent stem cells (iPSCs) give rise to HE with robust hemogenic potential, the generation of bona fide HSCs from iPSCs remains a challenge. Here, we map single cell dynamics of EHT during embryoid body differentiation from iPSCs and integrate it with human embryo datasets to identify key transcriptional differences between in vitro and in vivo cell states. We further map ligand-receptor interactions associated with differential expression of developmental programs in the iPSC system. We found that the expression of endothelial genes was incompletely repressed during iPSC EHT. Elevated FGF signaling by FGF23, an endothelial pathway ligand, was associated with differential gene expression between in vitro and in vivo EHT. Chemical inhibition of FGF signaling during EHT increased HSPC generation in the zebrafish, while an FGF agonist had the opposite effect. Consistently, chemical inhibition of FGF signaling increased hematopoietic output from iPSCs. In summary, we map the dynamics of EHT from iPSCs at single cell resolution and identify ligand-receptor interactions that can be modulated to improve iPSC differentiation protocols. We show, as proof of principle, that chemical inhibition of FGF signaling during EHT improves hematopoietic output in zebrafish and the iPSC system.
]]></description>
<dc:creator>Wellington, R.</dc:creator>
<dc:creator>Cheng, X.</dc:creator>
<dc:creator>Campbell, C. A.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Espin-Palazon, R.</dc:creator>
<dc:creator>Hadland, B.</dc:creator>
<dc:creator>Doulatov, S.</dc:creator>
<dc:date>2024-09-24</dc:date>
<dc:identifier>doi:10.1101/2024.09.24.612755</dc:identifier>
<dc:title><![CDATA[Developmental regulation of endothelial-to-hematopoietic transition from induced pluripotent stem cells]]></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/10.1101/2024.09.24.614677v1?rss=1">
<title>
<![CDATA[
Recognition of phylogenetically diverse pathogens through enzymatically amplified recruitment of RNF213 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.24.614677v1?rss=1</link>
<description><![CDATA[
Innate immunity senses microbial ligands known as pathogen-associated molecular patterns (PAMPs). Except for nucleic acids, PAMPs are exceedingly taxa-specific, thus enabling pattern recognition receptors to detect cognate pathogens while ignoring others. How the E3 ubiquitin ligase RNF213 can respond to phylogenetically distant pathogens, including Gram-negative Salmonella, Gram-positive Listeria, and eukaryotic Toxoplasma, remains unknown. Here we report that the evolutionary history of RNF213 is indicative of repeated adaptation to diverse pathogen target structures, especially in and around its newly identified CBM20 carbohydrate-binding domain, which we have resolved by cryo-EM. We find that RNF213 forms coats on phylogenetically distant pathogens. ATP hydrolysis by RNF213s dynein-like domain is essential for coat formation on all three pathogens studied as is RZ finger-mediated E3 ligase activity for bacteria. Coat formation is not diffusion-limited but instead relies on rate-limiting initiation events and subsequent cooperative incorporation of further RNF213 molecules. We conclude that RNF213 responds to evolutionarily distant pathogens through enzymatically amplified cooperative recruitment.
]]></description>
<dc:creator>Crespillo-Casado, A.</dc:creator>
<dc:creator>Pothukuchi, P.</dc:creator>
<dc:creator>Naydenova, K.</dc:creator>
<dc:creator>Yip, M. C. J.</dc:creator>
<dc:creator>Young, J. M.</dc:creator>
<dc:creator>Boulanger, J.</dc:creator>
<dc:creator>Dharamdasani, V.</dc:creator>
<dc:creator>Harper, C.</dc:creator>
<dc:creator>Hammoudi, P.-M.</dc:creator>
<dc:creator>Otten, E. G.</dc:creator>
<dc:creator>Boyle, K. B.</dc:creator>
<dc:creator>Gogoi, M.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:creator>Randow, F.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.24.614677</dc:identifier>
<dc:title><![CDATA[Recognition of phylogenetically diverse pathogens through enzymatically amplified recruitment of RNF213]]></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/10.1101/2024.09.25.615007v1?rss=1">
<title>
<![CDATA[
Multimodal Spatial Profiling Reveals Immune Suppression and Microenvironment Remodeling in Fallopian Tube Precursors to High-Grade Serous Ovarian Carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.25.615007v1?rss=1</link>
<description><![CDATA[
High-Grade Serous Ovarian Cancer (HGSOC) originates from fallopian tube (FT) precursors. However, the molecular changes that occur as precancerous lesions progress to HGSOC are not well understood. To address this, we integrated high-plex imaging and spatial transcriptomics to analyze human tissue samples at different stages of HGSOC development, including p53 signatures, serous tubal intraepithelial carcinomas (STIC), and invasive HGSOC. Our findings reveal immune modulating mechanisms within precursor epithelium, characterized by chromosomal instability, persistent interferon (IFN) signaling, and dysregulated innate and adaptive immunity. FT precursors display elevated expression of MHC-class I, including HLA-E, and IFN-stimulated genes, typically linked to later-stage tumorigenesis. These molecular alterations coincide with progressive shifts in the tumor microenvironment, transitioning from immune surveillance in early STICs to immune suppression in advanced STICs and cancer. These insights identify potential biomarkers and therapeutic targets for HGSOC interception and clarify the molecular transitions from precancer to cancer.

STATEMENT OF SIGNIFICANCEThis study maps the immune response in fallopian tube precursors of high-grade serous ovarian cancer, highlighting localized interferon signaling, CIN, and competing immune surveillance and suppression along the progression axis. It provides an explorable public spatial profiling atlas for investigating precancer mechanisms, biomarkers, and early detection and interception strategies.
]]></description>
<dc:creator>Kader, T.</dc:creator>
<dc:creator>Lin, J.-R.</dc:creator>
<dc:creator>Hug, C.</dc:creator>
<dc:creator>Coy, S.</dc:creator>
<dc:creator>Chen, Y.-A.</dc:creator>
<dc:creator>de Bruijn, I.</dc:creator>
<dc:creator>Shih, N.</dc:creator>
<dc:creator>Jung, E.</dc:creator>
<dc:creator>Pelletier, R. J.</dc:creator>
<dc:creator>Leon, M. L.</dc:creator>
<dc:creator>Mingo, G.</dc:creator>
<dc:creator>Omran, D. K.</dc:creator>
<dc:creator>Lee, J. S.</dc:creator>
<dc:creator>Yapp, C.</dc:creator>
<dc:creator>Satravada, B. A.</dc:creator>
<dc:creator>Kundra, R.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Chan, S.</dc:creator>
<dc:creator>Tefft, J.</dc:creator>
<dc:creator>Muhlich, J. L.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Gysler, S. M.</dc:creator>
<dc:creator>Agudo, J.</dc:creator>
<dc:creator>Heath, J. R.</dc:creator>
<dc:creator>Schultz, N.</dc:creator>
<dc:creator>Drescher, C.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>Drapkin, R.</dc:creator>
<dc:creator>Santagata, S.</dc:creator>
<dc:date>2024-09-27</dc:date>
<dc:identifier>doi:10.1101/2024.09.25.615007</dc:identifier>
<dc:title><![CDATA[Multimodal Spatial Profiling Reveals Immune Suppression and Microenvironment Remodeling in Fallopian Tube Precursors to High-Grade Serous Ovarian Carcinoma]]></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/10.1101/2024.09.26.615177v1?rss=1">
<title>
<![CDATA[
Endocrine persistence in ER+ breast cancer is accompanied by metabolic vulnerability in oxidative phosphorylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.26.615177v1?rss=1</link>
<description><![CDATA[
Despite adjuvant treatment with endocrine therapies, estrogen receptor-positive (ER+) breast cancers recur in a significant proportion of patients. Recurrences are attributable to clinically undetectable endocrine-tolerant persister cancer cells that retain tumor-forming potential. Therefore, strategies targeting such persister cells may prevent recurrent disease. Using CRISPR-Cas9 genome-wide knockout screening in ER+ breast cancer cells, we identified a survival mechanism involving metabolic reprogramming with reliance upon mitochondrial respiration in endocrine-tolerant persister cells. Quantitative proteomic profiling showed reduced levels of glycolytic proteins in persisters. Metabolic tracing of glucose revealed an energy-depleted state in persisters where oxidative phosphorylation was required to generate ATP. A phase II clinical trial was conducted to evaluate changes in mitochondrial markers in primary ER+/HER2-breast tumors induced by neoadjuvant endocrine therapy (NCT04568616). In an analysis of tumor specimens from 32 patients, tumors exhibiting residual cell proliferation after aromatase inhibitor-induced estrogen deprivation with letrozole showed increased mitochondrial content. Genetic profiling and barcode lineage tracing showed that endocrine-tolerant persistence occurred stochastically without genetic predisposition. Mice bearing cell line- and patient-derived xenografts were used to measure the anti-tumor effects of mitochondrial complex I inhibition in the context of endocrine therapy. Pharmacological inhibition of complex I suppressed the tumor-forming potential of persisters and synergized with the anti-estrogen fulvestrant to induce regression of patient-derived xenografts. These findings indicate that mitochondrial metabolism is essential in endocrine-tolerant persister ER+ breast cancer cells and warrant the development of treatment strategies to leverage this vulnerability in the context of endocrine-sensitive disease.

Statement of SignificanceEndocrine-tolerant persister cancer cells that survive endocrine therapy can cause recurrent disease. Persister cells exhibit increased energetic dependence upon mitochondria for survival and tumor re-growth potential.
]]></description>
<dc:creator>Tau, S.</dc:creator>
<dc:creator>Chamberlin, M. D.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Marotti, J. D.</dc:creator>
<dc:creator>Roberts, A. M.</dc:creator>
<dc:creator>Carmichael, M. M.</dc:creator>
<dc:creator>Cressey, L.</dc:creator>
<dc:creator>Dragnev, C. P.</dc:creator>
<dc:creator>Demidenko, E.</dc:creator>
<dc:creator>Hampsch, R. A.</dc:creator>
<dc:creator>Soucy, S. M.</dc:creator>
<dc:creator>Kolling, F. W.</dc:creator>
<dc:creator>Samkoe, K. S.</dc:creator>
<dc:creator>Alvarez, J. V.</dc:creator>
<dc:creator>Kettenbach, A.</dc:creator>
<dc:creator>Miller, T. W.</dc:creator>
<dc:date>2024-09-27</dc:date>
<dc:identifier>doi:10.1101/2024.09.26.615177</dc:identifier>
<dc:title><![CDATA[Endocrine persistence in ER+ breast cancer is accompanied by metabolic vulnerability in oxidative phosphorylation]]></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/10.1101/2024.09.27.615528v1?rss=1">
<title>
<![CDATA[
Damage-induced IL-18 stimulates thymic NK Cells limiting endogenous tissue regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.27.615528v1?rss=1</link>
<description><![CDATA[
Interleukin-18 is an acute phase pro-inflammatory molecule crucial for mediating viral clearance by activating Th1 CD4+, cytotoxic CD8+ T, and NK cells. Here, we show that mature IL-18 is generated in the thymus following numerous distinct forms of tissue damage, all of which cause caspase-1-mediated immunogenic cell death. We report that IL-18 stimulated cytotoxic NK cells limit endogenous thymic regeneration, a critical process that ensures restoration of immune competence after acute insults like stress, infection, chemotherapy, and radiation. NK cells suppressed thymus recovery by aberrantly targeting thymic epithelial cells (TECs), which act as the master regulators of organ function and regeneration. Together these studies reveal a novel pathway regulating tissue regeneration in the thymus and offer IL-18 as a potential therapeutic target to boost thymic function. Moreover, given the enthusiasm for IL-18 as a cancer immunotherapy for its capacity to elicit a type-1 immune response, these findings also offer insight into potential off-target effects.
]]></description>
<dc:creator>Granadier, D.</dc:creator>
<dc:creator>Cooper, K.</dc:creator>
<dc:creator>Kousa, A.</dc:creator>
<dc:creator>Acenas, D.</dc:creator>
<dc:creator>Lemarquis, A.</dc:creator>
<dc:creator>Hernandez, V.</dc:creator>
<dc:creator>Warren, M.</dc:creator>
<dc:creator>Iovino, L.</dc:creator>
<dc:creator>deRoos, P.</dc:creator>
<dc:creator>Lederer, E.</dc:creator>
<dc:creator>Shannon-Sevillano, S.</dc:creator>
<dc:creator>Kinsella, S.</dc:creator>
<dc:creator>Evandy, C.</dc:creator>
<dc:creator>van den Brink, M.</dc:creator>
<dc:creator>Dudakov, J.</dc:creator>
<dc:date>2024-09-30</dc:date>
<dc:identifier>doi:10.1101/2024.09.27.615528</dc:identifier>
<dc:title><![CDATA[Damage-induced IL-18 stimulates thymic NK Cells limiting endogenous tissue regeneration]]></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/10.1101/2024.10.03.616425v1?rss=1">
<title>
<![CDATA[
Conserved chromatin regulators control the transcriptional immune response to intracellular pathogens in Caenorhabditis elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.03.616425v1?rss=1</link>
<description><![CDATA[
Robust transcriptional responses are critical for defense against infection. However, unrestrained immune responses can cause negative impacts such as damaging inflammation and slowed development. Here we find that a class of transcriptional regulators previously associated with regulation of development in Caenorhabditis elegans, is also involved in immune responses. Specifically, through forward genetics, we find that loss of lin-15B leads to constitutive expression of Intracellular Pathogen Response (IPR) genes. lin-15B encodes a transcriptional repressor with a conserved THAP domain that is associated with the DRM chromatin remodeling complex that regulates C. elegans development. We show that lin-15B mutants have increased resistance to natural intracellular pathogens, and the induction of IPR genes in lin-15B mutants relies on the MES-4 histone methyltransferase. We extend our analyses to other DRM and NuRD chromatin remodeling factors, as well as SUMOylation histone modifiers, showing that a broad range of chromatin-related factors can repress IPR gene expression. Altogether these findings suggest that conserved chromatin regulators may facilitate development in part by repressing damaging immune responses against intracellular pathogens.

AUTHOR SUMMARYIn this study, we show that transcriptional regulators, previously linked to development in C. elegans, also control immune responses. Through forward genetic screens, we found that loss of LIN-15B leads to constitutive activation of Intracellular Pathogen Response (IPR) genes. LIN-15B is part of the DREAM chromatin remodeling complex, and its loss enhances resistance to intracellular pathogens. This immune response depends on the MES-4 histone methyltransferase. We also discovered that other chromatin regulators, including NuRD and SUMOylation factors, similarly repress IPR gene expression, highlighting a new role in immunity for these conserved regulators of development.
]]></description>
<dc:creator>Tecle, E.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Blanchard, M. J.</dc:creator>
<dc:creator>Bui, T.</dc:creator>
<dc:creator>Chhan, C. B.</dc:creator>
<dc:creator>Underwood, R. S.</dc:creator>
<dc:creator>Bakowski, M. A.</dc:creator>
<dc:creator>Troemel, E. R.</dc:creator>
<dc:creator>Lazetic, V.</dc:creator>
<dc:date>2024-10-04</dc:date>
<dc:identifier>doi:10.1101/2024.10.03.616425</dc:identifier>
<dc:title><![CDATA[Conserved chromatin regulators control the transcriptional immune response to intracellular pathogens in Caenorhabditis elegans]]></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/10.1101/2024.10.10.617484v1?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/10.1101/2024.10.10.616351v1?rss=1">
<title>
<![CDATA[
Computational Microbiome Pharmacology Analysis Elucidates the Anti-Cancer Potential of Vaginal Microbes and Metabolites 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.10.616351v1?rss=1</link>
<description><![CDATA[
The vaginal microbiomes role in risk, progression, and treatment of female cancers has been widely explored. Yet, there remains a need to develop methods to understand the interaction of microbiome factors with host cells and to characterize their potential therapeutic functions. To address this challenge, we developed a systems biology framework we term the Pharmacobiome for microbiome pharmacology analysis. The Pharmacobiome framework evaluates similarities between microbes and microbial byproducts and known drugs based on their impact on host transcriptomic cellular signatures. Here, we apply our framework to characterization of the Anti-Gynecologic Cancer Vaginal Pharmacobiome. Using published vaginal microbiome multi-omics data from the Partners PrEP clinical trial, we constructed vaginal epithelial gene signatures associated with each profiled vaginal microbe and metabolite. We compared these microbiome-associated host gene signatures to post-drug perturbation host gene signatures associated with 35 FDA-approved anti-cancer drugs from the Library of Integrated Network-based Cellular Signatures database to identify vaginal microbes and metabolites with high statistical and functional similarity to these drugs. We found that Lactobacilli and their metabolites can regulate host gene expression in ways similar to many anti-cancer drugs. Additionally, we experimentally tested our model prediction that taurine, a metabolite produced by L. crispatus, kills cancerous breast and endometrial cancer cells. Our study shows that the Pharmacobiome is a powerful framework for characterizing the anti-cancer therapeutic potential of vaginal microbiome factors with generalizability to other cancers, microbiomes, and diseases.
]]></description>
<dc:creator>Lawore, D. C.</dc:creator>
<dc:creator>Jena, S.</dc:creator>
<dc:creator>Berard, A. R.</dc:creator>
<dc:creator>Birse, K.</dc:creator>
<dc:creator>Lamont, A.</dc:creator>
<dc:creator>Mackelprang, R. D.</dc:creator>
<dc:creator>Noel-Romas, L.</dc:creator>
<dc:creator>Perner, M.</dc:creator>
<dc:creator>Hou, X.</dc:creator>
<dc:creator>Irungu, E.</dc:creator>
<dc:creator>Mugo, N.</dc:creator>
<dc:creator>Knodel, S.</dc:creator>
<dc:creator>Muwonge, T. R.</dc:creator>
<dc:creator>Katabira, E.</dc:creator>
<dc:creator>Hughes, S. M.</dc:creator>
<dc:creator>Levy, C.</dc:creator>
<dc:creator>Calienes, F. L.</dc:creator>
<dc:creator>Baeten, J. M.</dc:creator>
<dc:creator>Celum, C.</dc:creator>
<dc:creator>Hladik, F.</dc:creator>
<dc:creator>Lingappa, J. R.</dc:creator>
<dc:creator>Burgener, A. D.</dc:creator>
<dc:creator>Green, L. N.</dc:creator>
<dc:creator>Brubaker, D. K.</dc:creator>
<dc:date>2024-10-11</dc:date>
<dc:identifier>doi:10.1101/2024.10.10.616351</dc:identifier>
<dc:title><![CDATA[Computational Microbiome Pharmacology Analysis Elucidates the Anti-Cancer Potential of Vaginal Microbes and Metabolites]]></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/10.1101/2024.10.13.618066v1?rss=1">
<title>
<![CDATA[
SUMOylation differentially regulates SMCHD1 complex formation and function in a genomic context-specific manner 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.13.618066v1?rss=1</link>
<description><![CDATA[
Structural Maintenance of Chromosomes Hinge Domain Containing 1 (SMCHD1) is a chromatin repressor regulating gene expression and chromatin architecture of distinct autosomal and X-chromosomal loci. SMCHD1 mutations cause derepression of the D4Z4 macrosatellite repeat-embedded DUX4 gene in skeletal muscle of facioscapulohumeral muscular dystrophy (FSHD) patients Little is known about the regulation and post-translational modification of SMCHD1. Here we report the SUMOylation dynamics of SMCHD1 and its impact on autosomal single copy and repetitive loci, and the inactive X chromosome (Xi).

We identify that SMCHD1 is SUMOylated primarily at lysine 1374, uncover factors regulating SMCHD1 SUMOylation, and demonstrate that SMCHD1 interacts with chromatin repressors TRIM28, HNRNPK and SETDB1 in a SUMO-dependent manner. We find that SUMOylation impacts Xi engagement of SMCHD1, maintenance of a repressive D4Z4 chromatin structure preventing DUX4 expression, and regulation of LRIF1 promoter activity. The rapid, SUMO-dependent upregulation of DUX4 could explain the bursts of DUX4 expression typical for FSHD muscle.
]]></description>
<dc:creator>Goossens, R.</dc:creator>
<dc:creator>Tihaya, M. S.</dc:creator>
<dc:creator>Willemsen, I. M.</dc:creator>
<dc:creator>Straasheijm, K. R.</dc:creator>
<dc:creator>van der Vliet, P. J.</dc:creator>
<dc:creator>Gonzalez-Prieto, R.</dc:creator>
<dc:creator>Vertegaal, A. C. O.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:creator>Balog, J.</dc:creator>
<dc:creator>van der Maarel, S. M.</dc:creator>
<dc:date>2024-10-13</dc:date>
<dc:identifier>doi:10.1101/2024.10.13.618066</dc:identifier>
<dc:title><![CDATA[SUMOylation differentially regulates SMCHD1 complex formation and function in a genomic context-specific manner]]></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/10.1101/2024.10.14.617881v1?rss=1">
<title>
<![CDATA[
Sugar-rich foods exacerbate antibiotic-induced microbiome injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.14.617881v1?rss=1</link>
<description><![CDATA[
Intestinal microbiota composition is implicated in several diseases; understanding the factors that influence it are key to elucidating host-commensal interactions and to designing microbiome-targeted therapies. We quantified how diet influences microbiome dynamics in hospitalized patients. We recorded 9,419 meals consumed by 173 patients undergoing hematopoietic cell transplantation and profiled the microbiome in 1,009 longitudinally collected stool samples from 158 of them. Caloric intake was correlated with fecal microbiota diversity. Bayesian inference revealed associations between intake of sweets or sugars during antibiotic exposure with microbiome disruption, as assessed by low diversity or expansion of the pathobiont Enterococcus. We validated this observation experimentally, finding that sucrose exacerbated antibiotic-induced Enterococcus expansion in mice. Taken together, our results suggest that avoiding sugar-rich foods during antibiotic treatment may reduce microbiome injury.

One Sentence SummaryAnalyses of hematopoietic cell transplant patients and of mice uncover links between foods and microbiome disruption.
]]></description>
<dc:creator>Dai, A.</dc:creator>
<dc:creator>Adintori, P. A.</dc:creator>
<dc:creator>Funnell, T.</dc:creator>
<dc:creator>Jogia, W. P.</dc:creator>
<dc:creator>Fei, T.</dc:creator>
<dc:creator>Waters, N. R.</dc:creator>
<dc:creator>Rangesa, M.</dc:creator>
<dc:creator>Ballweg, A.</dc:creator>
<dc:creator>Gipson, B.</dc:creator>
<dc:creator>Raj, S.</dc:creator>
<dc:creator>Hayase, E.</dc:creator>
<dc:creator>Markey, K. A.</dc:creator>
<dc:creator>Burgos da Silva, M.</dc:creator>
<dc:creator>Miltiadous, O.</dc:creator>
<dc:creator>Brambilla, C. Z.</dc:creator>
<dc:creator>Buchan, M. L.</dc:creator>
<dc:creator>Peets, T.</dc:creator>
<dc:creator>Gradissimo, A.</dc:creator>
<dc:creator>Smith, N.</dc:creator>
<dc:creator>Katsamakis, Z.</dc:creator>
<dc:creator>Warren, A.</dc:creator>
<dc:creator>Amoretti, L. A.</dc:creator>
<dc:creator>Duan, C.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Matheis, F.</dc:creator>
<dc:creator>Sullivan, A. P.</dc:creator>
<dc:creator>Slingerland, J. B.</dc:creator>
<dc:creator>Clurman, A.</dc:creator>
<dc:creator>Brereton, D. G.</dc:creator>
<dc:creator>Giardina, P. A.</dc:creator>
<dc:creator>Gomes, A. L. C.</dc:creator>
<dc:creator>Johnson, A. J.</dc:creator>
<dc:creator>Knights, D.</dc:creator>
<dc:creator>Jenq, R. R.</dc:creator>
<dc:creator>Perales, M.-A.</dc:creator>
<dc:creator>Giralt, S. A.</dc:creator>
<dc:creator>Schluter, J.</dc:creator>
<dc:creator>van den Brink, M. R. M.</dc:creator>
<dc:creator>Peled, J.</dc:creator>
<dc:date>2024-10-17</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.617881</dc:identifier>
<dc:title><![CDATA[Sugar-rich foods exacerbate antibiotic-induced microbiome injury]]></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/10.1101/2024.10.14.618178v1?rss=1">
<title>
<![CDATA[
A Comprehensive Engineering Strategy Improves Potency and Manufacturability of a Near Pan-neutralizing Antibody Against HIV 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.14.618178v1?rss=1</link>
<description><![CDATA[
Anti-HIV envelope broadly neutralizing antibodies (bnAbs) are alternatives to conventional antiretrovirals with the potential to prevent and treat infection, reduce latent reservoirs, and/or mediate a functional cure. Clinical trials with "first generation" bnAbs used alone or in combination show promising antiviral effects but also highlight that additional engineering of "enhanced" antibodies will be required for optimal clinical utility, while preserving or enhancing cGMP manufacturing capability. Here we report the engineering of an anti-CD4 binding-site (CD4bs) bnAb, N49P9.3, purified from the plasma of an HIV elite-neutralizer. Through a series of rational modifications we produced a variant that demonstrates: enhanced potency; superior antiviral activity in combination with other bnAbs; low polyreactivity; and longer circulating half-life. Additional engineering for manufacturing produced a final variant, eN49P9, with properties conducive to cGMP production. Overall, these efforts demonstrate the feasibility of developing enhanced anti-CD4bs bnAbs with greatly improved antiviral properties as well as potential translational value.
]]></description>
<dc:creator>Sajadi, M.</dc:creator>
<dc:creator>Abbasi, A.</dc:creator>
<dc:creator>Rikhtegaran Tehrani, Z.</dc:creator>
<dc:creator>Siska, C.</dc:creator>
<dc:creator>Clark, R.</dc:creator>
<dc:creator>Chi, W.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Mielke, D.</dc:creator>
<dc:creator>Wagh, K.</dc:creator>
<dc:creator>Liu, Q.</dc:creator>
<dc:creator>Jumpa, T.</dc:creator>
<dc:creator>Ketchem, R. R.</dc:creator>
<dc:creator>Nguyen, D. N.</dc:creator>
<dc:creator>Tolbert, W. D.</dc:creator>
<dc:creator>Pierce, B. G.</dc:creator>
<dc:creator>Atkinson, B.</dc:creator>
<dc:creator>Deming, D.</dc:creator>
<dc:creator>Sprague, M.</dc:creator>
<dc:creator>Asakawa, A.</dc:creator>
<dc:creator>Ferrer, D.</dc:creator>
<dc:creator>Dunn, Y.</dc:creator>
<dc:creator>Calvillo, S.</dc:creator>
<dc:creator>Yin, R.</dc:creator>
<dc:creator>Guest, J. D.</dc:creator>
<dc:creator>Korber, B.</dc:creator>
<dc:creator>Mayer, B. T.</dc:creator>
<dc:creator>Sato, A. H.</dc:creator>
<dc:creator>Ouyang, X.</dc:creator>
<dc:creator>Foulke, S.</dc:creator>
<dc:creator>Habibzadeh, P.</dc:creator>
<dc:creator>Karimi, M.</dc:creator>
<dc:creator>Aslanabadi, A.</dc:creator>
<dc:creator>Hojabri, M.</dc:creator>
<dc:creator>Saadat, S.</dc:creator>
<dc:creator>Zareidoodeji, R.</dc:creator>
<dc:creator>Kedzior, M.</dc:creator>
<dc:creator>Pozharski, E.</dc:creator>
<dc:creator>Heredia, A.</dc:creator>
<dc:creator>Montefiori, D.</dc:creator>
<dc:creator>Ferrari, G.</dc:creator>
<dc:creator>Pazgier, M.</dc:creator>
<dc:creator>Lewis, G. K.</dc:creator>
<dc:creator>Jardine, J. G.</dc:creator>
<dc:creator>Lusso, P.</dc:creator>
<dc:creator>Devico, A.</dc:creator>
<dc:date>2024-10-17</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618178</dc:identifier>
<dc:title><![CDATA[A Comprehensive Engineering Strategy Improves Potency and Manufacturability of a Near Pan-neutralizing Antibody Against HIV]]></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/10.1101/2024.10.16.618753v1?rss=1">
<title>
<![CDATA[
Statistical analysis of repertoire data demonstrates the influence of microhomology in V(D)J recombination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.16.618753v1?rss=1</link>
<description><![CDATA[
V(D)J recombination generates the diverse B and T cell receptors essential for recognizing a wide array of antigens. This diversity arises from the combinatorial assembly of V(D)J genes and the junctional deletion and insertion of nucleotides. While previous in vitro studies have shown that microhomology---short stretches of sequence homology between gene ends---can bias the recombination process, the extent of microhomologys impact in vivo, particularly in humans, remains unknown. In this paper, we assess how germline-encoded microhomology influences trimming and ligation during V(D)J recombination using statistical inference on previously-published high-throughput TCR repertoire sequencing data. We find that microhomology increases both trimming and ligation probabilities, making it an important predictor of recombination outcomes. These effects are consistent across different receptor loci and sequence types. Further, we demonstrate that accounting for microhomology effects significantly alters sequence annotation probabilities and rankings, highlighting its practical importance for accurately inferring the V(D)J recombination events that generated an observed sequence. Together, these results enhance our understanding of how microhomologous nucleotides shape the human V(D)J recombination process.

Significance StatementHumans rely on diverse adaptive immune receptor repertoires to effectively defend against infections. The receptor generation process, known as V(D)J recombination, is designed to create this diversity by stochastically joining V(D)J gene segments and modifying their junctions through nucleotide deletions and insertions. Previous studies, conducted in vitro, have suggested that short stretches of homologous nucleotides between gene segments can bias these recombination steps. In this study, we explore the extent to which these homologous nucleotides influence V(D)J recombination in humans using statistical inference on large-scale receptor repertoire sequencing data. Our findings reveal that microhomology significantly biases several recombination steps, with important practical implications for the analysis, processing, and interpretation of receptor sequences.
]]></description>
<dc:creator>Russell, M. L.</dc:creator>
<dc:creator>Trofimov, A.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2024-10-18</dc:date>
<dc:identifier>doi:10.1101/2024.10.16.618753</dc:identifier>
<dc:title><![CDATA[Statistical analysis of repertoire data demonstrates the influence of microhomology in V(D)J recombination]]></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/10.1101/2024.10.17.618956v1?rss=1">
<title>
<![CDATA[
Protein catabolites as blood-based biomarkers of aging physiology: Findings from the Dog Aging Project 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.17.618956v1?rss=1</link>
<description><![CDATA[
Our understanding of age-related physiology and metabolism has grown through the study of systems biology, including transcriptomics, single-cell analysis, proteomics and metabolomics. Studies in lab organisms in controlled environments, while powerful and complex, fall short of capturing the breadth of genetic and environmental variation in nature. Thus, there is now a major effort in geroscience to identify aging biomarkers and to develop aging interventions that might be applied across the diversity of humans and other free-living species. To meet this challenge, the Dog Aging Project (DAP) is designed to identify cross-sectional and longitudinal patterns of aging in complex systems, and how these are shaped by the diversity of genetic and environmental variation among companion dogs. Here we surveyed the plasma metabolome from the first year of sampling of the Precision Cohort of the DAP. By incorporating extensive metadata and whole genome sequencing information, we were able to overcome the limitations inherent in breed-based estimates of genetic and physiological effects, and to probe the physiological and dietary basis of the age-related metabolome. We identified a significant effect of age on approximately 40% of measured metabolites. Among other insights, we discovered a potentially novel biomarker of age in the post-translationally modified amino acids (ptmAAs). The ptmAAs, which can only be generated by protein hydrolysis, covaried both with age and with other biomarkers of amino acid metabolism, and in a way that was robust to diet. Clinical measures of kidney function mediated about half of the higher ptmAA levels in older dogs. This work identifies ptmAAs as robust indicators of age in dogs, and points to kidney function as a physiological mediator of age-associated variation in the plasma metabolome.
]]></description>
<dc:creator>Harrison, B. R.</dc:creator>
<dc:creator>Partida-Aguilar, M.</dc:creator>
<dc:creator>Marye, A.</dc:creator>
<dc:creator>Djukovic, D.</dc:creator>
<dc:creator>Kauffman, M.</dc:creator>
<dc:creator>Dunbar, M. D.</dc:creator>
<dc:creator>Mariner, B. L.</dc:creator>
<dc:creator>McCoy, B. M.</dc:creator>
<dc:creator>Algavi, Y. M.</dc:creator>
<dc:creator>Muller, E.</dc:creator>
<dc:creator>Baum, S.</dc:creator>
<dc:creator>Bamberger, T.</dc:creator>
<dc:creator>Raftery, D.</dc:creator>
<dc:creator>Creevy, K. E.</dc:creator>
<dc:creator>Dog Aging Project Consortium,</dc:creator>
<dc:creator>Avery, A.</dc:creator>
<dc:creator>Borenstein, E.</dc:creator>
<dc:creator>Snyder-Mackler, N.</dc:creator>
<dc:creator>Promislow, D. E.</dc:creator>
<dc:date>2024-10-21</dc:date>
<dc:identifier>doi:10.1101/2024.10.17.618956</dc:identifier>
<dc:title><![CDATA[Protein catabolites as blood-based biomarkers of aging physiology: Findings from the Dog Aging Project]]></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/10.1101/2024.10.21.619495v1?rss=1">
<title>
<![CDATA[
Transcriptomic landscape identifies two unrecognized ependymoma subtypes and novel pathways in medulloblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.21.619495v1?rss=1</link>
<description><![CDATA[
Medulloblastoma and ependymoma are common pediatric central nervous system tumors with significant molecular and clinical heterogeneity. We collected bulk RNA sequencing data from 888 medulloblastoma and 370 ependymoma tumors to establish a comprehensive reference landscape. Following rigorous batch effect correction, normalization, and dimensionality reduction, we constructed a unified landscape to explore gene expression, signaling pathways, RNA fusions, and copy number variations. Our analysis revealed distinct clustering patterns, including two primary ependymoma compartments, EPN-E1 and EPN-E2, each with specific RNA fusions and molecular signatures. In medulloblastoma, we observed precise stratification of Group 3/4 tumors by subtype and in SHH tumors by patient age. This landscape serves as a vital resource for identifying biomarkers, refining diagnoses, and enables the mapping of new patients bulk RNA-seq data onto the reference framework to predict biology and outcome from nearest neighbor analysis facilitate accurate disease subtype identification. The landscape is accessible via Oncoscape, an interactive platform, empowering global exploration and application.

One Sentence SummaryA landscape built using only Transcriptomic analysis for medulloblastoma and ependymoma reveals novel insights about subtype-specific biology.
]]></description>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Holland, E.</dc:creator>
<dc:creator>Vardharajan, S.</dc:creator>
<dc:creator>Taylor, M.</dc:creator>
<dc:creator>Sahm, F.</dc:creator>
<dc:creator>Mack, S. C.</dc:creator>
<dc:creator>Sievers, P.</dc:creator>
<dc:creator>Korshunov, A.</dc:creator>
<dc:creator>Nuechterlein, N.</dc:creator>
<dc:creator>Jensen, M.</dc:creator>
<dc:creator>Glatzer, G.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.21.619495</dc:identifier>
<dc:title><![CDATA[Transcriptomic landscape identifies two unrecognized ependymoma subtypes and novel pathways in medulloblastoma]]></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/10.1101/2024.10.25.620266v1?rss=1">
<title>
<![CDATA[
Long-term hybridization in a karst window reveals the genetic basis of eye loss in cavefish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.25.620266v1?rss=1</link>
<description><![CDATA[
Eye loss is a hallmark trait of animals inhabiting perpetual darkness, yet the precise genetic variants underlying this evolutionary change remain largely unknown. The Mexican tetra (Astyanax mexicanus) provides a powerful model for dissecting the genetic basis of eye degeneration, as sighted surface fish and multiple independently evolved blind cave populations remain interfertile; yet despite decades of research and numerous QTL studies, the genetic basis of eye loss has remained unresolved at the level of specific variants. Here, we exploit a rare natural experiment in the Caballo Moro cave, where the collapse of a karst window created a partially illuminated pool inhabited by both fully eyed and completely eyeless cavefish of closely related genetic background. Whole-genome sequencing reveals a long-standing hybrid population between cave and surface lineages, enabling a dramatic refinement of the genetic architecture of eye degeneration to 203 candidate SNPs across 41 genes. Among these, we identified a nonsynonymous mutation in the lens gap-junction protein Connexin-50 (Cx50). CRISPR-based disruption of cx50 induces early eye loss in surface fish, and F2 laboratory crosses confirm genetic linkage between cx50 variants and eye size. Additional Cx50 mutations are present in independent cavefish populations and correlate with reduced eye size. Notably, variants in conserved regions of Cx50 also occur in other cave-dwelling fish and subterranean mammals, suggesting repeated evolutionary targeting of this gene. Introduction of the Caballo Moro mutation into mice causes cataracts and reduced eye and lens size, confirming its functional impact. Together, these findings identify the first SNP directly implicated in cavefish eye loss, demonstrate the power of natural hybrid populations to resolve the genetic basis of complex traits, and reveal Cx50 as a case of molecular convergence in vertebrate eye degeneration.
]]></description>
<dc:creator>Kellermeyer, R.</dc:creator>
<dc:creator>Seidel, C.</dc:creator>
<dc:creator>Redwine, W. B.</dc:creator>
<dc:creator>Moran, R. L.</dc:creator>
<dc:creator>Bertho, S.</dc:creator>
<dc:creator>Ornelas-Garcia, C. P.</dc:creator>
<dc:creator>Alegre, D.</dc:creator>
<dc:creator>Weaver, K. J.</dc:creator>
<dc:creator>Unruh, J.</dc:creator>
<dc:creator>Troutwine, B.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Collins, E.</dc:creator>
<dc:creator>Rutkowski, J.</dc:creator>
<dc:creator>McGaugh, S. E.</dc:creator>
<dc:creator>Espinasa, L.</dc:creator>
<dc:creator>Rohner, N.</dc:creator>
<dc:date>2024-10-28</dc:date>
<dc:identifier>doi:10.1101/2024.10.25.620266</dc:identifier>
<dc:title><![CDATA[Long-term hybridization in a karst window reveals the genetic basis of eye loss in cavefish]]></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/10.1101/2024.11.04.621904v1?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/10.1101/2024.11.07.622561v1?rss=1">
<title>
<![CDATA[
MOSAIC: A Pipeline for MicrobiOme Studies Analytical Integration and Correction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.07.622561v1?rss=1</link>
<description><![CDATA[
Large-scale and consortium microbiome studies have enabled identification of reliable population-level biomedical signals, wherein integration is essential to eliminate unwanted variations between batches or studies and retain biological signals. Many strategies, each with distinct advantages and limitations, have been adapted or developed for microbiome data. The optimal strategy for a given study needs to be determined on a data-specific, case-by-case basis. Here, we develop the first-of-its-kind MicrobiOme Studies Analytical Integration and Correction (MOSAIC) pipeline to enable a convenient, fair, and comprehensive comparison of integration strategies. It includes modules for pre-processing, integration, and evaluation of artifact removal and signal preservation, using metrics relevant to common microbiome analyses, including alpha and beta diversities, disease prediction, and differential abundance analysis. We applied MOSAIC to extensive real-world and simulated data and found that though no single strategy excels in all aspects, yet certain strategies, the ComBat and ConQuR families, perform better overall.
]]></description>
<dc:creator>Fu, C.</dc:creator>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Ling, W.</dc:creator>
<dc:date>2024-11-11</dc:date>
<dc:identifier>doi:10.1101/2024.11.07.622561</dc:identifier>
<dc:title><![CDATA[MOSAIC: A Pipeline for MicrobiOme Studies Analytical Integration and Correction]]></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/10.1101/2024.11.10.622859v1?rss=1">
<title>
<![CDATA[
ROSIE: AI generation of multiplex immunofluorescence staining from histopathology images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.10.622859v1?rss=1</link>
<description><![CDATA[
Hematoxylin and eosin (H&E) is a common and inexpensive histopathology assay. Though widely used and information-rich, it cannot directly inform about specific molecular markers, which require additional experiments to assess. To address this gap, we present ROSIE, a deep-learning framework that computationally imputes the expression and localization of dozens of proteins from H&E images. Our model is trained on a dataset of over 1000 paired and aligned H&E and multiplex immunofluorescence (mIF) samples from 20 tissues and disease conditions, spanning over 16 million cells. Validation of our in silico mIF staining method on held-out H&E samples demonstrates that the predicted biomarkers are effective in identifying cell phenotypes, particularly distinguishing lymphocytes such as B cells and T cells, which are not readily discernible with H&E staining alone. Additionally, ROSIE facilitates the robust identification of stromal and epithelial microenvironments and immune cell subtypes like tumor-infiltrating lymphocytes (TILs), which are important for understanding tumor-immune interactions and can help inform treatment strategies in cancer research.
]]></description>
<dc:creator>Wu, E.</dc:creator>
<dc:creator>Bieniosek, M.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Thakkar, N.</dc:creator>
<dc:creator>Charville, G. W.</dc:creator>
<dc:creator>Makky, A.</dc:creator>
<dc:creator>Schürch, C.</dc:creator>
<dc:creator>Huyghe, J. R.</dc:creator>
<dc:creator>Peters, U.</dc:creator>
<dc:creator>Li, C. I.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Giba, H.</dc:creator>
<dc:creator>Behera, V.</dc:creator>
<dc:creator>Raman, A.</dc:creator>
<dc:creator>Trevino, A. E.</dc:creator>
<dc:creator>Mayer, A. T.</dc:creator>
<dc:creator>Zou, J.</dc:creator>
<dc:date>2024-11-11</dc:date>
<dc:identifier>doi:10.1101/2024.11.10.622859</dc:identifier>
<dc:title><![CDATA[ROSIE: AI generation of multiplex immunofluorescence staining from histopathology images]]></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/10.1101/2024.11.11.623044v1?rss=1">
<title>
<![CDATA[
Mutant RIT1 cooperates with YAP to drive an EMT-like lung cancer state 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.11.623044v1?rss=1</link>
<description><![CDATA[
The discovery of oncogene addiction in cancer has led to the development of over a dozen FDA-approved biomarker-driven therapies in lung adenocarcinoma. Somatic mutations of the "Ras-like in all tissues" (RIT1) gene are non-canonical driver events in lung cancer, occurring in [~]2% of lung adenocarcinomas in a mutually exclusive fashion with KRAS and EGFR mutations. Patients with RIT1-mutant lung cancer lack targeted therapy treatment options, and a lack of pre-clinical models has hindered the development of therapeutic strategies for RIT1-mutant lung cancer. Here we report a new mouse model of RIT1-driven lung cancer in which the human RIT1M90I variant can be induced in a Cre-regulated manner. We show that autochthonous expression of RIT1M90I in the lung weakly promotes cancer alone or in combination with loss of the p53 tumor suppressor. However, potent synergy between RIT1M90I and inactivation of Nf2 drives an aggressive epithelial-to-mesenchymal (EMT) lung cancer with 100% penetrance and short latency. We show this oncogenic cooperation is driven by synergistic activation of cJUN, a component of the AP-1 complex. Therapeutic inhibition of MEK and YAP/TEAD suppressed RIT1-driven lung cancer in vivo. These data identify YAP/TEAD as an important mediator of RIT1s oncogenic potential and nominate TEAD as an important drug target in RIT1-mutant lung cancer.

HIGHLIGHTSO_LIWe report a new RIT1M90I-mutant autochthonous lung tumor model
C_LIO_LIThe most common oncogenic variant of RIT1, RIT1M90I, weakly promotes lung tumor development
C_LIO_LIRIT1M90I drives the formation of lethal lung tumors in cooperation with p53 and Nf2 tumor suppressor gene loss
C_LIO_LIRIT1M90I and YAP cooperatively regulate cJUN expression
C_LIO_LITherapeutic MEK and TEAD targeting suppresses RIT1M90I-driven tumorigenesis
C_LI
]]></description>
<dc:creator>Rominger, M. C.</dc:creator>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Moorthi, S.</dc:creator>
<dc:creator>McSharry, M.</dc:creator>
<dc:creator>Kamlapurkar, S.</dc:creator>
<dc:creator>O'Brien, S.</dc:creator>
<dc:creator>Waldum, A.</dc:creator>
<dc:creator>Lo, A.</dc:creator>
<dc:creator>Duke, F.</dc:creator>
<dc:creator>Lowe, A.</dc:creator>
<dc:creator>Cromwell, E.</dc:creator>
<dc:creator>Glabman, R.</dc:creator>
<dc:creator>Koehne, A.</dc:creator>
<dc:creator>Berger, A.</dc:creator>
<dc:date>2024-11-12</dc:date>
<dc:identifier>doi:10.1101/2024.11.11.623044</dc:identifier>
<dc:title><![CDATA[Mutant RIT1 cooperates with YAP to drive an EMT-like lung cancer state]]></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/10.1101/2024.11.12.622506v1?rss=1">
<title>
<![CDATA[
Semaphorin 4A maintains functional diversity of the hematopoietic stem cell pool 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.12.622506v1?rss=1</link>
<description><![CDATA[
Somatic stem cell pools are comprised of diverse, highly specialized subsets whose individual contribution is critical for the overall regenerative function. In the bone marrow, myeloid-biased HSC (myHSC) are indispensable for replenishment of myeloid cells and platelets during inflammatory response but at the same time, become irreversibly damaged during inflammation and aging. Here, we identify an extrinsic factor, Semaphorin 4A (Sema4A), which non cell-autonomously confers myHSC resilience to inflammatory stress. We show that the absence of Sema4A, myHSC inflammatory hyper-responsiveness in young mice drives excessive myHSC expansion, myeloid bias and profound loss of regenerative function with age. Mechanistically, Sema4A is mainly produced by neutrophils, signals via a cell surface receptor Plexin D1 and safeguards myHSC epigenetic state. Our study shows that by selectively protecting a distinct stem cell subset, an extrinsic factor preserves functional diversity of somatic stem cell pool throughout organismal lifespan.
]]></description>
<dc:creator>Toghani, D.</dc:creator>
<dc:creator>Gupte, S.</dc:creator>
<dc:creator>Zeng, S.</dc:creator>
<dc:creator>Mahammadov, E.</dc:creator>
<dc:creator>Crosse, E.</dc:creator>
<dc:creator>Seyedhassantehrani, N.</dc:creator>
<dc:creator>Burns, C.</dc:creator>
<dc:creator>Gravano, D.</dc:creator>
<dc:creator>Radtke, S.</dc:creator>
<dc:creator>Kiem, H.-P.</dc:creator>
<dc:creator>Rodriguez, S.</dc:creator>
<dc:creator>Carlesso, N.</dc:creator>
<dc:creator>Pradeep, A.</dc:creator>
<dc:creator>Georgiades, A.</dc:creator>
<dc:creator>Lucas, F.</dc:creator>
<dc:creator>Wilson, N. K.</dc:creator>
<dc:creator>Kinston, S. J.</dc:creator>
<dc:creator>Gottgens, B.</dc:creator>
<dc:creator>Zong, L.</dc:creator>
<dc:creator>Beerman, I.</dc:creator>
<dc:creator>Park, B.</dc:creator>
<dc:creator>Janssens, D.</dc:creator>
<dc:creator>Jones, D.</dc:creator>
<dc:creator>Toghani, A.</dc:creator>
<dc:creator>Nerlov, C.</dc:creator>
<dc:creator>Pietras, E. C.</dc:creator>
<dc:creator>Mesnieres, M.</dc:creator>
<dc:creator>Maes, C.</dc:creator>
<dc:creator>Kumanogoh, A.</dc:creator>
<dc:creator>Worzfeld, T.</dc:creator>
<dc:creator>Cheong, J.-G.</dc:creator>
<dc:creator>Josefowicz, S. Z.</dc:creator>
<dc:creator>Kharchenko, P.</dc:creator>
<dc:creator>Scadden, D.</dc:creator>
<dc:creator>Scialdone, A.</dc:creator>
<dc:creator>Spencer, J. A.</dc:creator>
<dc:creator>Silberstein, L.</dc:creator>
<dc:date>2024-11-13</dc:date>
<dc:identifier>doi:10.1101/2024.11.12.622506</dc:identifier>
<dc:title><![CDATA[Semaphorin 4A maintains functional diversity of the hematopoietic stem cell pool]]></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/10.1101/2024.11.18.624054v1?rss=1">
<title>
<![CDATA[
Exploration of the Roles of HLAs When Predicting Infection Status by T Cell Receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.18.624054v1?rss=1</link>
<description><![CDATA[
T cells are critical components of human immune system. When a cell is infected by a virus, it presents viral peptides on its surface using human leukocyte antigen (HLA) proteins. These peptide-HLA complexes are recognized by T cells through interactions with T cell receptors (TCRs). A human blood sample can contain millions of unique TCRs, which is a sample from the TCR repertoire of the individual. TCR repertoire-wide association studies (TReWAS) aim to evaluate the association between individual TCRs and disease or exposure status. Previous studies have demonstrated that TCRs associated with viral infections can be identified through TReWAS, and such TCRs can be used to predict current or past infection with high accuracy. Notably, many TCRs are strongly associated with specific HLA alleles, suggesting that incorporating HLA information could enhance the accuracy of TReWAS analyses and TCR-based predictions. In our study, we evaluated TCR-based predictions by conditioning on individual HLA alleles or their k-nearest neighbors. We observed improved prediction accuracy for certain HLA alleles. Furthermore, these HLA-specific predictions provide insights into the role of specific HLAs in the infection or disease of interest, offering potential applications in personalized medicine.
]]></description>
<dc:creator>Ding, F.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2024-11-21</dc:date>
<dc:identifier>doi:10.1101/2024.11.18.624054</dc:identifier>
<dc:title><![CDATA[Exploration of the Roles of HLAs When Predicting Infection Status by T Cell Receptors]]></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/10.1101/2024.11.21.624751v1?rss=1">
<title>
<![CDATA[
PFAS Compounds PFOA and Gen X are Teratogenic to Sea Urchin Embryos 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.21.624751v1?rss=1</link>
<description><![CDATA[
Per-and polyfluorinated substances (PFAS) are synthetic chemicals that are used to make fluoropolymer coatings found in many products, such as non-stick pans, clothing, cosmetics, and food packaging. These highly persistent molecules are known as "forever chemicals" since they neither degrade environmentally nor break down enzymatically within biological systems. PFAS compounds readily contaminate water sources, and as a result, certain PFAS molecules have bioaccumulated in exposed species including humans. The purpose of this study was to define the effect of two PFAS molecules, the ostensibly more toxic perfluorooctanoic acid (PFOA) and the more recent, reportedly safer chemical hexafluoropropylene oxide dimer acid (Gen X), on the development of Lytechinus variegatus sea urchin embryos. We examined the effects of PFOA and Gen X on development and patterning using morphological analysis, immunostaining, HCR-FISH, and Particle Image Velocimetry (PIV). The results show that both PFAS compounds are teratogenic to sea urchin embryos. PFOA and Gen X each function at different intervals during development and provoke distinct phenotypic and gene expression outcomes. Despite beliefs that Gen X would be a safer alternative, our findings indicate that Gen X has earlier and more severe effects on endomesoderm and dorsal-ventral axis specification, neural development and function, and pattern formation compared to PFOA. These results illustrate the dangerous teratogenic potential of environmentally accumulating PFAS like Gen X, underscoring the negative ecological implications that accompany continuing commercial and industrial use of PFAS in the absence of remediation strategies.

HIGHLIGHTSO_LIGen X is more teratogenic in sea urchins than PFOA
C_LIO_LIGen X acts earlier than PFOA and perturbs axial and germ layer specification
C_LIO_LIEach PFAS reduces neural numbers and perturbs ciliary behavior and swimming
C_LI
]]></description>
<dc:creator>Lion, A. T.</dc:creator>
<dc:creator>Bodine, S. M.</dc:creator>
<dc:creator>McCutcheon, K. R.</dc:creator>
<dc:creator>Ghogale, M.</dc:creator>
<dc:creator>Chandragiri, S.</dc:creator>
<dc:creator>Abayawardena, D.</dc:creator>
<dc:creator>Shrestha, B. D.</dc:creator>
<dc:creator>Descoteaux, A. E.</dc:creator>
<dc:creator>Alvarez, K.</dc:creator>
<dc:creator>Balkman, J. A.</dc:creator>
<dc:creator>Cocke, B.</dc:creator>
<dc:creator>Wikramanayake, A.</dc:creator>
<dc:creator>Schlezinger, J.</dc:creator>
<dc:creator>Wong, J. Y.</dc:creator>
<dc:creator>Prakash, V. N.</dc:creator>
<dc:creator>Bradham, C.</dc:creator>
<dc:date>2024-11-21</dc:date>
<dc:identifier>doi:10.1101/2024.11.21.624751</dc:identifier>
<dc:title><![CDATA[PFAS Compounds PFOA and Gen X are Teratogenic to Sea Urchin Embryos]]></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/10.1101/2024.11.20.624598v1?rss=1">
<title>
<![CDATA[
Improving antibody-mediated protection against HSV infection by eliminating interactions with the viral Fc receptor gE/gI 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.20.624598v1?rss=1</link>
<description><![CDATA[
Herpes simplex virus (HSV) encodes surface glycoproteins that are host defense evasion molecules, allowing the virus to escape immune clearance. In addition to their role in neuropathogenesis and cell-cell spread, glycoproteins E and I (gE/gI) form a viral Fc receptor (vFcR) for most subclasses and allotypes of human IgG and promote evasion of humoral immune responses. While monoclonal antibodies (mAbs) protect mice from neonatal HSV (nHSV) infections, the impact of the vFcR on mAb-mediated protection by binding to IgG is unknown. Using HSV-1 with intact and ablated gE-mediated IgG Fc binding, and Fc-engineered antibodies with modified ability to interact with gE/gI, we investigated the role of the vFcR in viral pathogenesis and mAb-mediated protection from nHSV. The gD-specific human mAb HSV8 modified to lack binding to gE exhibited enhanced neutralization and in vivo protection compared to its native IgG1 form. This improved protection by the engineered mAbs was dependent on the presence of the vFcR. Human IgG3 allotypes lacking vFcR binding also exhibited enhanced antiviral activity in vivo, suggesting that vaccines that robustly induce IgG3 responses could show enhanced protection. suggesting the value of vaccination strategies that robustly induce this subclass. Lastly, analysis of longitudinal responses to acute primary genital infection in humans raised the possibility that unlike most viruses, HSV may exhibited slow induction of IgG3. In summary, this study demonstrates that mAbs lacking the ability to interact with the vFcR can exhibit improved protection from HSV--offering new prospects for antibody-based interventions.

One Sentence Summary: The herpes simplex virus neutralizing antibody HSV8 demonstrates improved activity in vitro and in vivo when its IgG Fc domain lacks the ability to bind the viral Fc receptor glycoprotein E/I complex through either Fc engineering or natural human IgG3 allotypes.
]]></description>
<dc:creator>Slein, M. D.</dc:creator>
<dc:creator>Backes, I. M.</dc:creator>
<dc:creator>Kelkar, N. S.</dc:creator>
<dc:creator>Garland, C. R.</dc:creator>
<dc:creator>Khanwalkar, U. S.</dc:creator>
<dc:creator>Sholukh, A. M.</dc:creator>
<dc:creator>Johnston, C. M.</dc:creator>
<dc:creator>Leib, D. A.</dc:creator>
<dc:creator>Ackerman, M. E.</dc:creator>
<dc:date>2024-11-22</dc:date>
<dc:identifier>doi:10.1101/2024.11.20.624598</dc:identifier>
<dc:title><![CDATA[Improving antibody-mediated protection against HSV infection by eliminating interactions with the viral Fc receptor gE/gI]]></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/10.1101/2024.11.22.624910v1?rss=1">
<title>
<![CDATA[
Improved Deep Learning Prediction of TCR-HLA Association 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.22.624910v1?rss=1</link>
<description><![CDATA[
Understanding the relationship between T cell receptors (TCRs) and human leukocyte antigens (HLAs) is essential for elucidating immune response specificity, uncovering mechanisms of autoimmunity, and advancing targeted immunotherapies. We have previously developed a deep learning method, DePTH (Deep Learning Prediction of TCR-HLA associations), to predict the association between a TCR and an HLA based on their amino acid sequences. In this work, we evaluated the performance of DePTH in two additional datasets, and investigated the influence of two potential confounding factors: TCR generation probability and the sequence length of CDR3 (Complementarity-Determining Region 3), which is a key region in the antigen-binding site of TCRs. Building on these insights, we combined training data from two datasets to train a new version of DePTH: DePTH 2.0.
]]></description>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2024-11-25</dc:date>
<dc:identifier>doi:10.1101/2024.11.22.624910</dc:identifier>
<dc:title><![CDATA[Improved Deep Learning Prediction of TCR-HLA Association]]></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/10.1101/2024.11.26.625333v1?rss=1">
<title>
<![CDATA[
Association between COVID-19 Disease Severity and T Cell Receptor Repertoire 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625333v1?rss=1</link>
<description><![CDATA[
During the COVID-19 pandemic, while most infected individuals experienced mild to moderate symptoms, a significant subset developed severe illness. A clinical test distinguishing between mild and severe cases could inform effective treatment strategies. Toward the latter stages of the pandemic, it became evident that vaccination or prior infection cannot entirely prevent reinfection. However, they are crucial in reducing the risk of severe disease by inducing T-cell memory. T cell receptors (TCRs), which can be obtained from human blood, serve as valuable biomarkers for monitoring T cell responses to SARS-CoV-2 infection. In this study, we investigated the associations between TCR metrics and COVID-19 severity and found significant associations. Furthermore, such associations could depend on the subset of TCRs used (e.g., TCRs from CD8+ T or CD4+ T cells) and when the TCRs were collected.
]]></description>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Dai, C.</dc:creator>
<dc:creator>Goldman, J. D.</dc:creator>
<dc:creator>Heath, J. R.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2024-11-26</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625333</dc:identifier>
<dc:title><![CDATA[Association between COVID-19 Disease Severity and T Cell Receptor Repertoire]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.26.625407v1?rss=1">
<title>
<![CDATA[
Thrifty wide-context models of B cell receptor somatic hypermutation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625407v1?rss=1</link>
<description><![CDATA[
Somatic hypermutation (SHM) is the diversity-generating process in antibody affinity maturation. Probabilistic models of SHM are needed for analyzing rare mutations, for understanding the selective forces guiding affinity maturation, and for understanding the underlying biochemical process. High throughput data offers the potential to develop and fit models of SHM on relevant data sets. In this paper we model SHM using modern frameworks. We are motivated by recent work suggesting the importance of a wider context for SHM, however, assigning an independent rate to each k-mer leads to an exponential proliferation of parameters. Thus, using convolutions on 3-mer embeddings, we develop "thrifty" models of SHM of various sizes; these can have fewer free parameters than a 5-mer model and yet have a significantly wider context. These offer a slight performance improvement over a 5-mer model, and other modern model elaborations worsen performance. We also find that a per-site effect is not necessary to explain SHM patterns given nucleotide context. Also, the two current methods for fitting an SHM model -- on out-of-frame sequence data and on synonymous mutations -- produce significantly different results, and augmenting out-of-frame data with synonymous mutations does not aid out-of-sample performance.
]]></description>
<dc:creator>Sung, K.</dc:creator>
<dc:creator>Johnson, M. M.</dc:creator>
<dc:creator>Dumm, W.</dc:creator>
<dc:creator>Simon, N.</dc:creator>
<dc:creator>Haddox, H.</dc:creator>
<dc:creator>Fukuyama, J.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2024-12-01</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625407</dc:identifier>
<dc:title><![CDATA[Thrifty wide-context models of B cell receptor somatic hypermutation]]></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/10.1101/2024.11.29.624972v1?rss=1">
<title>
<![CDATA[
Two distinct durable human class-switched memory B cell populations are induced by vaccination and infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.29.624972v1?rss=1</link>
<description><![CDATA[
Memory lymphocytes are durable cells that persist in the absence of antigen, but few human B cell subsets have been characterized in terms of durability. The relative durability of eight non-overlapping human B cell sub-populations covering 100% of all human class-switched B cells was interrogated. Only two long-lived B cell populations persisted in the relative absence of antigen. In addition to canonical germinal center-derived switched-memory B cells with an IgD-CD27+ CXCR5+ phenotype, a second, non-canonical, but distinct memory population of IgD-CD27- CXCR5+ DN1 B cells was also durable, exhibited a unique TP63-linked transcriptional and anti-apoptotic signature, had low levels of somatic hypermutation, but was more clonally expanded than canonical switched-memory B cells. DN1 B cells likely evolved to preserve immunological breadth and may represent the human counterparts of rodent extrafollicular memory B cells that, unlike canonical memory B cells, can enter germinal centers and facilitate B cell and antibody evolution.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=140 SRC="FIGDIR/small/624972v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Perugino, C.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Feldman, J.</dc:creator>
<dc:creator>Marbourg, J.</dc:creator>
<dc:creator>Guy, T.</dc:creator>
<dc:creator>Hui, A.</dc:creator>
<dc:creator>Ingram, N.</dc:creator>
<dc:creator>Liebaert, J.</dc:creator>
<dc:creator>Chaudhary, N.</dc:creator>
<dc:creator>Tao, W.</dc:creator>
<dc:creator>Jacob-Dolan, C.</dc:creator>
<dc:creator>Hauser, B. M.</dc:creator>
<dc:creator>Mian, Z.</dc:creator>
<dc:creator>Nathan, A.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Kaseke, C.</dc:creator>
<dc:creator>Tano-Menka, R.</dc:creator>
<dc:creator>Getz, M.</dc:creator>
<dc:creator>Senjobe, F.</dc:creator>
<dc:creator>Berrios, C.</dc:creator>
<dc:creator>Ofoman, O.</dc:creator>
<dc:creator>Manickas-Hill, Z.</dc:creator>
<dc:creator>Wesemann, D.</dc:creator>
<dc:creator>Lemieux, J.</dc:creator>
<dc:creator>Goldberg, M. B.</dc:creator>
<dc:creator>Nundel, K.</dc:creator>
<dc:creator>Moormann, A. M.</dc:creator>
<dc:creator>Marshak-Rothstein, A.</dc:creator>
<dc:creator>Larocque, R.</dc:creator>
<dc:creator>Ryan, E.</dc:creator>
<dc:creator>Iafrate, A. J.</dc:creator>
<dc:creator>Lingwood, D.</dc:creator>
<dc:creator>Gaiha, G.</dc:creator>
<dc:creator>Charles, R.</dc:creator>
<dc:creator>Balazs, A. B.</dc:creator>
<dc:creator>Pandit, A.</dc:creator>
<dc:creator>Naranbhai, V.</dc:creator>
<dc:creator>Schmidt, A.</dc:creator>
<dc:creator>Pillai, S.</dc:creator>
<dc:date>2024-12-04</dc:date>
<dc:identifier>doi:10.1101/2024.11.29.624972</dc:identifier>
<dc:title><![CDATA[Two distinct durable human class-switched memory B cell populations are induced by vaccination and infection]]></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/10.1101/2024.12.05.627024v1?rss=1">
<title>
<![CDATA[
E-box independent chromatin recruitment turns MYOD into a transcriptional repressor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.05.627024v1?rss=1</link>
<description><![CDATA[
MYOD is an E-box sequence-specific basic Helix-Loop-Helix (bHLH) transcriptional activator that, when expressed in non-muscle cells, induces nuclear reprogramming toward skeletal myogenesis by promoting chromatin accessibility at previously silent loci. Here, we report on the identification of a previously unrecognized property of MYOD as repressor of gene expression, via E-box-independent chromatin binding within accessible genomic elements, which invariably leads to reduced chromatin accessibility. MYOD-mediated repression requires the integrity of functional domains previously implicated in MYOD-mediated activation of gene expression. Repression of mitogen-and growth factor-responsive genes occurs through promoter binding and requires a highly conserved domain within the first helix. Repression of cell-of-origin/alternative lineage genes occurs via binding and decommissioning of distal regulatory elements, such as super-enhancers (SE), which requires the N-terminal activation domain as well as two chromatin-remodeling domains and leads to reduced strength of CTCF-mediated chromatin interactions. Surprisingly, MYOD-mediated chromatin compaction and repression of transcription do not associate with reduction of H3K27ac, the conventional histone mark of enhancer or promoter activation, but with reduced levels of the recently discovered histone H4 acetyl-methyl lysine modification (Kacme). These results extend MYOD biological properties beyond the current dogma that restricts MYOD function to a monotone transcriptional activator and reveal a previously unrecognized functional versatility arising from an alternative chromatin recruitment through E-box or non-E-box sequences. The E-box independent repression of gene expression by MYOD might provide a promiscuous mechanism to reduce chromatin accessibility and repress cell-of-origin/alternative lineage and growth factor/mitogen-responsive genes to safeguard the integrity of cell identity during muscle progenitor commitment toward the myogenic lineage.
]]></description>
<dc:creator>Nicoletti, C.</dc:creator>
<dc:creator>Massenet, J.</dc:creator>
<dc:creator>Pintado-Urbanc, A. P.</dc:creator>
<dc:creator>Connor, L. J.</dc:creator>
<dc:creator>Nicolau, M.</dc:creator>
<dc:creator>Sundar, S.</dc:creator>
<dc:creator>Xu, M.</dc:creator>
<dc:creator>Schmitt, A.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Fang, Z.</dc:creator>
<dc:creator>Chan, T. C. I.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:creator>Cheung, T. H.</dc:creator>
<dc:creator>Simon, M. D.</dc:creator>
<dc:creator>Caputo, L.</dc:creator>
<dc:creator>Puri, P. L.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.05.627024</dc:identifier>
<dc:title><![CDATA[E-box independent chromatin recruitment turns MYOD into a transcriptional repressor]]></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/10.1101/2024.12.08.627395v1?rss=1">
<title>
<![CDATA[
HIPSTR: highest independent posterior subtree reconstruction in TreeAnnotator X 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.08.627395v1?rss=1</link>
<description><![CDATA[
In Bayesian phylogenetic and phylodynamic studies it is common to summarise the posterior distribution of trees with a time-calibrated consensus phylogeny. While the maximum clade credibility (MCC) tree is often used for this purpose, we here show that a novel consensus tree method - the highest independent posterior subtree reconstruction, or HIPSTR - contains consistently higher supported clades over MCC. We also provide faster computational routines for estimating both consensus trees in an updated version of TreeAnnotator X, an open-source software program that summarizes the information from a sample of trees and returns many helpful statistics such as individual clade credibilities contained in the consensus tree. HIPSTR and MCC reconstructions on two Ebola virus and two SARS-CoV-2 data sets show that HIPSTR yields consensus trees that consistently contain clades with higher support compared to MCC trees. The MCC trees regularly fail to include several clades with very high posterior probability ([&ge;] 0.95) as well as a large number of clades with moderate to high posterior probability ([&ge;] 0.50), whereas HIPSTR achieves near-perfect performance in this respect. HIPSTR also exhibits favorable computational performance over MCC in TreeAnnotator X. Comparison to the recently developed CCD0-MAP algorithm yielded mixed results, and requires more in-depth exploration in follow-up studies. TreeAnnotator X - which is part of the BEAST X (v10.5.0) software package - is available at https://github.com/beast-dev/beast-mcmc/releases.
]]></description>
<dc:creator>Baele, G.</dc:creator>
<dc:creator>Carvalho, L. M.</dc:creator>
<dc:creator>Brusselmans, M.</dc:creator>
<dc:creator>Dudas, G.</dc:creator>
<dc:creator>Ji, X.</dc:creator>
<dc:creator>McCrone, J. T.</dc:creator>
<dc:creator>Lemey, P.</dc:creator>
<dc:creator>Suchard, M. A.</dc:creator>
<dc:creator>Rambaut, A.</dc:creator>
<dc:date>2024-12-10</dc:date>
<dc:identifier>doi:10.1101/2024.12.08.627395</dc:identifier>
<dc:title><![CDATA[HIPSTR: highest independent posterior subtree reconstruction in TreeAnnotator X]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.11.627911v1?rss=1">
<title>
<![CDATA[
Single-cell multi-omic analysis of fibrolamellar carcinoma reveals rewired cell-to-cell communication patterns and unique vulnerabilities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.11.627911v1?rss=1</link>
<description><![CDATA[
Fibrolamellar carcinoma (FLC) is a rare malignancy disproportionately affecting adolescents and young adults with no standard of care. FLC is characterized by thick stroma, which has long suggested an important role of the tumor microenvironment. Over the past decade, several studies have revealed aberrant markers and pathways in FLC. However, a significant drawback of these efforts is that they were conducted on bulk tumor samples. Consequently, identities and roles of distinct cell types within the tumor milieu, and the patterns of intercellular communication, have yet to be explored. In this study we unveil cell-type specific gene signatures, transcription factor networks, and super-enhancers in FLC using a multi-omics strategy that leverages both single-nucleus ATAC-seq and single-nucleus RNA-seq. We also infer completely rewired cell-to-cell communication patterns in FLC including signaling mediated by SPP1-CD44, MIF-ACKR3, GDF15-TGFBR2, and FGF7-FGFR. Finally, we validate findings with loss-of-function studies in several models including patient tissue slices, identifying vulnerabilities that merit further investigation as candidate therapeutic targets in FLC.
]]></description>
<dc:creator>Farghli, A. R.</dc:creator>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Sherman, M. S.</dc:creator>
<dc:creator>Dickerson, L. K.</dc:creator>
<dc:creator>Shui, B.</dc:creator>
<dc:creator>Nukaya, M.</dc:creator>
<dc:creator>Stephanou, A.</dc:creator>
<dc:creator>Ma, R. K.</dc:creator>
<dc:creator>Pepe-Mooney, B. J.</dc:creator>
<dc:creator>Smith, C. J.</dc:creator>
<dc:creator>Long, D.</dc:creator>
<dc:creator>Munn, P. R.</dc:creator>
<dc:creator>McNairn, A.</dc:creator>
<dc:creator>Grenier, J. K.</dc:creator>
<dc:creator>Karski, M.</dc:creator>
<dc:creator>Ronnekleiv-Kelly, S. M.</dc:creator>
<dc:creator>Pillarisetty, V. G.</dc:creator>
<dc:creator>Goessling, W.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:creator>Vakili, K.</dc:creator>
<dc:creator>Sethupathy, P.</dc:creator>
<dc:date>2024-12-12</dc:date>
<dc:identifier>doi:10.1101/2024.12.11.627911</dc:identifier>
<dc:title><![CDATA[Single-cell multi-omic analysis of fibrolamellar carcinoma reveals rewired cell-to-cell communication patterns and unique vulnerabilities]]></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/10.1101/2024.12.12.628092v1?rss=1">
<title>
<![CDATA[
Sir2 is required for the quiescence-specific condensed three-dimensional chromatin structure of rDNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.12.628092v1?rss=1</link>
<description><![CDATA[
Quiescence in Saccharomyces cerevisiae is a reversible G0 crucial for long-term survival under nutrient-deprived conditions. During quiescence, the genome is hypoacetylated and chromatin undergoes significant compaction. However, the 3D structure of the ribosomal DNA (rDNA) locus in this state is not well understood. Here, we report that the rDNA locus in quiescent cells forms a distinct condensed loop-like structure, different from structures observed during the mitotic cell cycle. Deletion of SIR2 disrupts this structure, causing it to collapse into a small dot and resulting in quiescence entry and exit defects. In contrast, Sir2 affects rDNA structure only modestly in G2/M phase. In the absence of Sir2, occupancy of both RNA Polymerase II and histone H3 increase at the rDNA locus during quiescence and through quiescence exit, further indicating gross defects in chromatin structure. Together, these results uncover a previously undescribed rDNA chromatin structure specific to quiescent cells and underscore the importance of Sir2 in facilitating the transition between cellular states.
]]></description>
<dc:creator>Cucinotta, C.</dc:creator>
<dc:creator>Dell, R.</dc:creator>
<dc:creator>Alavattam, K.</dc:creator>
<dc:creator>Tsukiyama, T.</dc:creator>
<dc:date>2024-12-12</dc:date>
<dc:identifier>doi:10.1101/2024.12.12.628092</dc:identifier>
<dc:title><![CDATA[Sir2 is required for the quiescence-specific condensed three-dimensional chromatin structure of rDNA]]></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/10.1101/2024.12.17.628988v1?rss=1">
<title>
<![CDATA[
DUX4-induced HSATII RNA accumulation drives protein aggregation impacting RNA processing pathways. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.17.628988v1?rss=1</link>
<description><![CDATA[
RNA-driven protein aggregation leads to cellular dysregulation, disrupting normal cellular processes, and contributing to the development of diseases and tumorigenesis. Here, we show that double homeobox 4 (DUX4), an early embryonic transcription factor and causative gene of facioscapulohumeral muscular dystrophy (FSHD), induces the accumulation of stable intranuclear RNAs, including nucleolar RNA and human satellite II (HSATII) RNA. Stable intranuclear RNAs drive protein aggregation in DUX4-expressing muscle cells. Specifically, HSATII RNA sequesters RNA methylation factors. HSATII-YBX1 ribonucleoprotein (RNP) complex formation is mediated by HSATII double-stranded RNA and NSUN2 activity. Aberrant HSATII-RNP complexes affect RNA processing pathways, including RNA splicing. Differential splicing of genes mediated by HSATII-RNP complexes are associated with pathways known to be dysregulated by DUX4 expression. These findings highlight the broader influence of DUX4 on nuclear RNA dynamics and suggest that HSATII RNA could be a critical mediator of RNA processing regulation. Understanding the impact of HSATII-RNP formation on RNA processing provides insight into the molecular mechanisms underlying FSHD.

SUMMARYArends et al. show that DUX4-induction of stable intranuclear RNA, including pericentromeric human satellite II (HSATII) repeat RNA, leads to nuclear protein aggregation. HSATII ribonucleoprotein complexes impact RNA processing downstream of DUX4 expression.
]]></description>
<dc:creator>Arends, T.</dc:creator>
<dc:creator>Bennett, S. R.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:date>2024-12-18</dc:date>
<dc:identifier>doi:10.1101/2024.12.17.628988</dc:identifier>
<dc:title><![CDATA[DUX4-induced HSATII RNA accumulation drives protein aggregation impacting RNA processing pathways.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.18.629070v1?rss=1">
<title>
<![CDATA[
Deciphering single-cell heterogeneity and cellular ecosystem dynamics during prostate cancer progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.629070v1?rss=1</link>
<description><![CDATA[
Prostate cancer (PC) progresses from benign epithelium through pre-malignant lesions, localized tumors, metastatic dissemination, and castration-resistant stages, with some cases exhibiting phenotype plasticity under therapeutic pressure. However, high-resolution insights into how cell phenotypes evolve across successive stages of PC remain limited. Here, we present the Prostate Cancer Cell Atlas (PCCAT) by integrating [~]710,000 single cells from 197 human samples covering a spectrum of tumor stages. This comprehensive analysis dissects the cellular landscape and characterizes key cell types and molecular features that associate with PC progression and prognosis. In malignant cells, we highlight a distinctive profile denoted by high Major Histocompatibility Complex (MHC) expression, low Androgen Receptor (AR) activity, and enhanced stemness programs associated with enzalutamide resistance. Moreover, we reveal several cell states strongly correlated with PC progression and adverse prognosis, including lineage plasticity-like malignant cells (LPCs), neuroendocrine tumor cells, pericytes, and matrix cancer-associated fibroblasts (mCAFs). Furthermore, we uncover shared cell states that underpin the immune suppressive tumor microenvironment in advanced PC, including activated regulatory T cells, exhausted CD8+ T cells, and SPP1-expressing macrophages. Lastly, we pinpoint a spatial niche composed of mCAFs and SPP1-expressing macrophages localized near the tumor boundary in aggressive PC, which correlates with poor prognosis. Overall, our work provides a valuable resource and offers deeper insights into the diverse cell states, dynamics, and functional characteristics involved in PC progression at single-cell resolution.
]]></description>
<dc:creator>Zhao, F.</dc:creator>
<dc:creator>Zeng, J.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Zhang, T.</dc:creator>
<dc:creator>Thomas, G. V.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Alumkal, J. J.</dc:creator>
<dc:creator>Moran, A. E.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629070</dc:identifier>
<dc:title><![CDATA[Deciphering single-cell heterogeneity and cellular ecosystem dynamics during prostate cancer progression]]></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/10.1101/2024.12.18.628976v1?rss=1">
<title>
<![CDATA[
Detecting Clinically Relevant Topological Structures in Multiplexed Spatial Proteomics Imaging Using TopKAT 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.628976v1?rss=1</link>
<description><![CDATA[
Novel multiplexed spatial proteomics imaging platforms expose the spatial architecture of cells in the tumor microenvironment (TME). The diverse cell population in the TME, including its spatial context, has been shown to have important clinical implications, correlating with disease prognosis and treatment response. The accelerating implementation of spatial proteomic technologies motivates new statistical models to test if cell-level images associate with patient-level endpoints. Few existing methods can robustly characterize the geometry of the spatial arrangement of cells and also yield both a valid and powerful test for association with patient-level outcomes. We propose a topology-based approach that combines persistent homology with kernel testing to determine if topological structures created by cells predict continuous, binary, or survival clinical endpoints. We term our method TopKAT (Topological Kernel Association Test) and show that it can be more powerful than statistical tests grounded in the spatial point process model, particularly when cells arise along the boundary of a ring. We demonstrate the properties of TopKAT through simulation studies and apply it to two studies of triple negative breast cancer where we show that TopKAT recovers clinically relevant topological structures in the spatial distribution of immune and tumor cells.
]]></description>
<dc:creator>Samorodnitsky, S.</dc:creator>
<dc:creator>Campbell, K. M.</dc:creator>
<dc:creator>Little, A.</dc:creator>
<dc:creator>Ling, W.</dc:creator>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Chen, Y.-C.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:date>2024-12-21</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.628976</dc:identifier>
<dc:title><![CDATA[Detecting Clinically Relevant Topological Structures in Multiplexed Spatial Proteomics Imaging Using TopKAT]]></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/10.1101/2024.12.22.629997v1?rss=1">
<title>
<![CDATA[
Probing mechanical selection in diverse eukaryotic genomes through accurate prediction of 3D DNA mechanics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.22.629997v1?rss=1</link>
<description><![CDATA[
Connections between the mechanical properties of DNA and biological functions have been speculative due to the lack of methods to measure or predict DNA mechanics at scale. Recently, a proxy for DNA mechanics, cyclizability, was measured by loop-seq and enabled genome-scale investigation of DNA mechanics. Here, we use this dataset to build a computational model predicting bias-corrected intrinsic cyclizability, with near-perfect accuracy, solely based on DNA sequence. Further, the model predicts intrinsic bending direction in 3D space. Using this tool, we aimed to probe mechanical selection - that is, the evolutionary selection of DNA sequence based on its mechanical properties - in diverse circumstances. First, we found that the intrinsic bend direction of DNA sequences correlated with the observed bending in known protein-DNA complex structures, suggesting that many proteins co-evolved with their DNA partners to capture DNA in its intrinsically preferred bent conformation. We then applied our model to large-scale yeast population genetics data and showed that centromere DNA element II, whose consensus sequence is unknown, leaving its sequence-specific role unclear, is under mechanical selection to increase the stability of inner-kinetochore structure and to facilitate centromeric histone recruitment. Finally, in silico evolution under strong mechanical selection discovered hallucinated sequences with cyclizability values so extreme that they required experimental validation, yet, found in nature in the densely packed mitochondrial(mt) DNA of Namystynia karyoxenos, an ocean-dwelling protist with extreme mitochondrial gene fragmentation. The need to transmit an extraordinarily large amount of mtDNA, estimated to be > 600 Mb, in combination with the absence of mtDNA compaction proteins may have pushed mechanical selection to the extreme. Similarly extreme DNA mechanics are observed in bird microchromosomes, although the functional consequence is not yet clear. The discovery of eccentric DNA mechanics in unrelated unicellular and multicellular eukaryotes suggests that we can predict extreme natural biology which can arise through strong selection. Our methods offer a way to study the biological functions of DNA mechanics in any genome and to engineer DNA sequences with desired mechanical properties.
]]></description>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Prokopchuk, G.</dc:creator>
<dc:creator>Popchock, A. R.</dc:creator>
<dc:creator>Hao, J.</dc:creator>
<dc:creator>Liao, T.-W.</dc:creator>
<dc:creator>Yan, S.</dc:creator>
<dc:creator>Hedman, D. J.</dc:creator>
<dc:creator>Larson, J. D.</dc:creator>
<dc:creator>Walther, B.</dc:creator>
<dc:creator>Becker, N. A.</dc:creator>
<dc:creator>Basu, A.</dc:creator>
<dc:creator>Maher, L. J.</dc:creator>
<dc:creator>Wheeler, R. J.</dc:creator>
<dc:creator>Asbury, C. L.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:creator>Lukes, J.</dc:creator>
<dc:creator>Ha, T.</dc:creator>
<dc:date>2024-12-23</dc:date>
<dc:identifier>doi:10.1101/2024.12.22.629997</dc:identifier>
<dc:title><![CDATA[Probing mechanical selection in diverse eukaryotic genomes through accurate prediction of 3D DNA mechanics]]></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/10.1101/2024.12.23.630206v1?rss=1">
<title>
<![CDATA[
Histone H4 limits transcription of the histone locus in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.23.630206v1?rss=1</link>
<description><![CDATA[
In all eukaryotes DNA replication is coupled to histone synthesis to coordinate chromatin packaging of the genome. Canonical histone genes coalesce in the nucleus into the Histone Locus Body (HLB), where gene transcription and 3 mRNA processing occurs. Both histone gene transcription and mRNA stability are reduced when DNA replication is inhibited, implying that the Histone Locus Body senses the rate of DNA synthesis. In Drosophila melanogaster, the S-phase-induced histone genes are tandemly repeated in an [~]100 copy array, whereas in humans, these histone genes are scattered. In both organisms these genes coalesce into Histone Locus Bodies. We used a transgenic histone gene reporter and RNAi in Drosophila to identify canonical H4 histone as a unique repressor of histone synthesis during the G2 phase in germline cells. Using cytology and CUT&Tag chromatin profiling, we find that histone H4 uniquely occupies histone gene promoters in both Drosophila and human cells. Our results suggest that repression of histone genes by soluble histone H4 is a conserved mechanism that coordinates DNA replication with histone synthesis in proliferating cells.
]]></description>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Wooten, M.</dc:creator>
<dc:creator>Takushi, B. N.</dc:creator>
<dc:creator>Vidaurre, V.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2024-12-24</dc:date>
<dc:identifier>doi:10.1101/2024.12.23.630206</dc:identifier>
<dc:title><![CDATA[Histone H4 limits transcription of the histone locus in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-24</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/10.1101/2025.01.07.631013v1?rss=1">
<title>
<![CDATA[
The mutation rate of SARS-CoV-2 is highly variable between sites and is influenced by sequence context, genomic region, and RNA structure 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.07.631013v1?rss=1</link>
<description><![CDATA[
RNA viruses like SARS-CoV-2 have a high mutation rate, which contributes to their rapid evolution. The rate of mutations depends on the mutation type (e.g., A[-&gt;]C, A[-&gt;]G, etc.) and can vary between sites in the viral genome. Understanding this variation can shed light on the mutational processes at play, and is crucial for quantitative modeling of viral evolution. Using the millions of available SARS-CoV-2 full-genome sequences, we estimate rates of synonymous mutations for all 12 possible nucleotide mutation types and examine how much these rates vary between sites. We find a surprisingly high level of variability and several striking patterns: the rates of four mutation types suddenly increase at one of two gene boundaries; the rates of most mutation types strongly depend on a sites local sequence context, with up to 56-fold differences between contexts; consistent with a previous study, the rates of some mutation types are lower at sites engaged in RNA secondary structure. A simple log-linear model of these features explains [~]15-60% of the fold-variation of mutation rates between sites, depending on mutation type; more complex models only modestly improve predictive power out of sample. We estimate the fitness effect of each mutation based on the number of times it actually occurs versus the number of times it is expected to occur based on the model. We identify several small regions of the genome where synonymous or noncoding mutations occur much less often than expected, indicative of strong purifying selection on the RNA sequence that is independent of protein sequence. Overall, this work expands our basic understanding of SARS-CoV-2s evolution by characterizing the viruss mutation process at the level of individual sites and uncovering several striking mutational patterns that arise from unknown mechanisms.
]]></description>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Angehrn, G.</dc:creator>
<dc:creator>Sesta, L.</dc:creator>
<dc:creator>Jennings-Shaffer, C.</dc:creator>
<dc:creator>Temple, S. D.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>DeWitt, W. S.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:date>2025-01-08</dc:date>
<dc:identifier>doi:10.1101/2025.01.07.631013</dc:identifier>
<dc:title><![CDATA[The mutation rate of SARS-CoV-2 is highly variable between sites and is influenced by sequence context, genomic region, and RNA structure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.07.631807v1?rss=1">
<title>
<![CDATA[
Metagenomic estimation of absolute bacterial biomass in the mammalian gut through host-derived read normalization. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.07.631807v1?rss=1</link>
<description><![CDATA[
Absolute bacterial biomass estimation in the human gut is crucial for understanding microbiome dynamics and host-microbe interactions. Current methods for quantifying bacterial biomass in stool, such as flow cytometry, qPCR, or spike-ins (i.e., adding cells or DNA from an organism not normally found in a sample), can be labor-intensive, costly, and confounded by factors like water content, DNA extraction efficiency, PCR inhibitors, and other technical challenges that add bias and noise. We propose a simple, cost-effective approach that circumvents some of these technical challenges: directly estimating bacterial biomass from metagenomes using bacterial-to-host (B:H) read ratios. We compare B:H ratios to the standard methods outlined above, demonstrating that B:H ratios are useful proxies for bacterial biomass in stool and possibly in other host-associated substrates. We show how B:H ratios can be used to track antibiotic treatment response and recovery in both mice and humans, which showed 403-fold and 45-fold reductions in bacterial biomass during antibiotic treatment, respectively. Our results indicate that host and bacterial metagenomic DNA fractions in human stool fluctuate longitudinally around a stable mean in healthy individuals, and the average host read fraction varies across healthy individuals by < 8-9 fold. B:H ratios offer a convenient alternative to other absolute biomass quantification methods, without the need for additional measurements, experimental design considerations, or machine learning algorithms, enabling retrospective absolute biomass estimates from existing stool metagenomic data.
]]></description>
<dc:creator>Tang, G.</dc:creator>
<dc:creator>Carr, A. V.</dc:creator>
<dc:creator>Perez, C.</dc:creator>
<dc:creator>Ramos Sarmiento, K.</dc:creator>
<dc:creator>Levy, L.</dc:creator>
<dc:creator>Lampe, J.</dc:creator>
<dc:creator>Diener, C.</dc:creator>
<dc:creator>Gibbons, S. M.</dc:creator>
<dc:date>2025-01-08</dc:date>
<dc:identifier>doi:10.1101/2025.01.07.631807</dc:identifier>
<dc:title><![CDATA[Metagenomic estimation of absolute bacterial biomass in the mammalian gut through host-derived read normalization.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.13.632690v1?rss=1">
<title>
<![CDATA[
A fast numerical integration scheme for clonal expansion processes on graphs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.13.632690v1?rss=1</link>
<description><![CDATA[
Compound birth-death processes are widely used to model the age-incidence curves of many cancers [1]. There are efficient schemes for directly computing the relevant probability distributions in the context of linear multi-stage clonal expansion (MSCE) models [2]. However, these schemes have not been generalised to models on arbitrary graphs, forcing the use of either full stochastic simulations or mean-field approximations, which can become inaccurate at late times or old ages [3, 4]. Here, we present a numerical integration scheme for directly computing survival probabilities of a first-order birth-death process on an arbitrary directed graph, without the use of stochastic simulations. As a concrete application, we show that this new numerical method can be used to infer the parameters of an example graphical model from simulated data.
]]></description>
<dc:creator>Paterson, C. G. B.</dc:creator>
<dc:creator>Gao, M.</dc:creator>
<dc:creator>Hellier, J.</dc:creator>
<dc:creator>Luebeck, G.</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>Bozic, I.</dc:creator>
<dc:date>2025-01-15</dc:date>
<dc:identifier>doi:10.1101/2025.01.13.632690</dc:identifier>
<dc:title><![CDATA[A fast numerical integration scheme for clonal expansion processes on graphs]]></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/10.1101/2025.01.13.632860v1?rss=1">
<title>
<![CDATA[
Combinatorial phenotypic landscape enables bacterial resistance to phage infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.13.632860v1?rss=1</link>
<description><![CDATA[
AbstractSuccess of phage therapies is limited by bacterial defenses against phages. While a large variety of anti- phage defense mechanisms has been characterized, how expression of these systems is distributed across individual cells and how their combined activities translate into protection from phages has not been studied. Using bacterial single-cell RNA sequencing, we profiled the transcriptomes of [~]50,000 cells from cultures of a human pathobiont, Bacteroides fragilis, infected with a lytic bacteriophage. We quantified the asynchronous progression of phage infection in single bacterial cells and reconstructed the infection timeline, characterizing both host and phage transcriptomic changes as infection unfolded. We discovered a subpopulation of bacteria that remained uninfected and determined the heterogeneously expressed host factors associated with protection. Each cells vulnerability to phage infection was defined by combinatorial phase-variable expression of multiple genetic loci, including capsular polysaccharide (CPS) biosynthesis pathways, restriction-modification systems (RM), and a previously uncharacterized operon likely encoding fimbrial genes. By acting together, these heterogeneously expressed phase-variable systems and anti-phage defense mechanisms create a phenotypic landscape where distinct protective combinations enable the survival and re-growth of bacteria expressing these phenotypes without acquiring additional mutations. The emerging model of complementary action of multiple protective mechanisms heterogeneously expressed across an isogenic bacterial population showcases the potent role of phase variation and stochasticity in bacterial anti-phage defenses.

One Sentence SummaryCombinatorial phenotypic states with differential vulnerability to phage infection across a Bacteroides fragilis population enable a small number of super-resistant bacterial cells to evade the phage without the need for acquiring mutations.
]]></description>
<dc:creator>Gupta, A.</dc:creator>
<dc:creator>Morella, N.</dc:creator>
<dc:creator>Sutormin, D.</dc:creator>
<dc:creator>Li, N.</dc:creator>
<dc:creator>Gaisser, K.</dc:creator>
<dc:creator>Robertson, A.</dc:creator>
<dc:creator>Ispolatov, Y.</dc:creator>
<dc:creator>Seelig, G.</dc:creator>
<dc:creator>Dey, N.</dc:creator>
<dc:creator>Kuchina, A.</dc:creator>
<dc:date>2025-01-15</dc:date>
<dc:identifier>doi:10.1101/2025.01.13.632860</dc:identifier>
<dc:title><![CDATA[Combinatorial phenotypic landscape enables bacterial resistance to phage infection]]></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/10.1101/2025.01.16.633453v1?rss=1">
<title>
<![CDATA[
Pretransplant targeting of TNFRSF25 and CD25 stimulates recipient Tregs in target tissues ameliorating GVHD post-HSCT 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.16.633453v1?rss=1</link>
<description><![CDATA[
The current approach to minimize transplant-associated complications, including graft-versus-host disease (GVHD) includes long-term pharmacological immune suppression frequently accompanied by unwanted side effects. Advances in targeted immunotherapies regulating alloantigen responses in the recipient continue to reduce the need for pan-immunosuppression. Here, in vivo targeting of the TNF superfamily receptor 25 (TNFRSF25) and the high affinity IL-2 receptor with a TL1A-Ig fusion protein and low dose IL-2, respectively, was used to pretreat recipient mice prior to allogeneic-HSCT (aHSCT). Pretreatment induced Treg expansion persisting early post-aHSCT leading to diminished GVHD and improved transplant outcomes. Expansion was accompanied by an increase in frequency of stable and functionally active Tregs as evidenced by in vitro assays using cells from major GVHD target tissues including colon, liver, and eye. Importantly, pretreatment supported epithelial cell function/integrity, a diverse microbiome including reduction of pathologic bacteria overgrowth and promotion of butyrate producing bacteria, while maintaining physiologic levels of obligate/facultative anaerobes. Notably, using a sphingosine 1-phosphate receptor agonist to sequester T cells in lymphoid tissues, we found that the increased tissue Treg frequency included resident CD69+CD103+FoxP3+ hepatic Tregs. In contrast to infusion of donor Treg cells, the strategy developed here resulted in the presence of immunosuppressive target tissue environments in the recipient prior to the receipt of donor allo-reactive T cells and successful perseveration of GVL responses. We posit strategies that circumvent the need of producing large numbers of ex-vivo manipulated Tregs, may be accomplished through in vivo recipient Treg expansion, providing translational approaches to improve aHSCT outcomes.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=101 SRC="FIGDIR/small/633453v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>McManus, D.</dc:creator>
<dc:creator>Copsel, S.</dc:creator>
<dc:creator>Pfieffer, B. J.</dc:creator>
<dc:creator>Wolf, D.</dc:creator>
<dc:creator>Barreras, H.</dc:creator>
<dc:creator>Ma, S.</dc:creator>
<dc:creator>Khodor, A.</dc:creator>
<dc:creator>Komai, S.</dc:creator>
<dc:creator>Burgos da Silva, M.</dc:creator>
<dc:creator>Hazime, H.</dc:creator>
<dc:creator>Gallardo, M.</dc:creator>
<dc:creator>van den Brink, M. R.</dc:creator>
<dc:creator>Abreu, M. T.</dc:creator>
<dc:creator>Hill, G. R.</dc:creator>
<dc:creator>Perez, V. L.</dc:creator>
<dc:creator>Levy, R. B.</dc:creator>
<dc:date>2025-01-21</dc:date>
<dc:identifier>doi:10.1101/2025.01.16.633453</dc:identifier>
<dc:title><![CDATA[Pretransplant targeting of TNFRSF25 and CD25 stimulates recipient Tregs in target tissues ameliorating GVHD post-HSCT]]></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/10.1101/2025.01.22.634135v1?rss=1">
<title>
<![CDATA[
Clonal interference and changing selective pressures shape the escape of SARS-CoV-2 from hundreds of antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.22.634135v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 rapidly evolves to evade human immunity. While the viruss overall resistance to human polyclonal antibody responses has steadily increased over time, the dynamics by which it escaped individual monoclonal antibodies within these responses have not been thoroughly explored. Recently, a series of studies by Cao et al. [1, 2, 3] used deep mutational scanning (DMS) to identify which mutations allow the Wuhan-Hu-1 receptor-binding domain to escape binding by individual antibodies, doing so for thousands of antibodies. Here, we sought to use these data to retrospectively examine the evolutionary dynamics of escape from a set of 1,603 antibodies. For each antibody, we used the DMS data to predict an antibody-escape score for each of thousands of globally circulating viral sequences from the first 3.5 years of the pandemic, and then computed an escape trajectory that quantifies how the populations average escape score changed over time. We use pseudovirus neutralization data from Cao et al. and Wang et al. [4] to validate common patterns in escape trajectories. While some trajectories increase monotonically over time, others show large fluctuations as a result of clade-displacement events that reduce the frequency of antibody-escape mutations in the viral population. Fitness effects of mutations estimated from natural sequences suggest that the mutations are displaced due to clonal interference. Further, these estimates suggest that the order in which escape mutations arose is shaped by changing selective pressures. Overall, this work helps describe how SARS-CoV-2 evaded the individual components of a polyclonal immune response in nature, and suggests that evasion occurred via complex evolutionary dynamics.
]]></description>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Abdel Aziz, O.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>Kent, J.</dc:creator>
<dc:creator>Cooper, C. R.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2025-01-24</dc:date>
<dc:identifier>doi:10.1101/2025.01.22.634135</dc:identifier>
<dc:title><![CDATA[Clonal interference and changing selective pressures shape the escape of SARS-CoV-2 from hundreds of antibodies]]></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/10.1101/2025.01.24.634737v1?rss=1">
<title>
<![CDATA[
Centromeres in the thermotolerant yeast K. marxianusmediate attachment to a single microtubule 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.24.634737v1?rss=1</link>
<description><![CDATA[
Eukaryotic chromosome segregation requires spindle microtubules to attach to chromosomes through kinetochores. The chromosomal locus that mediates kinetochore assembly is the centromere and is epigenetically specified in most organisms by a centromeric histone H3 variant called CENP-A. An exception to this is budding yeast which have short, sequenced-defined point centromeres. In S. cerevisiae, a single CENP-A nucleosome is formed at the centromere and is sufficient for kinetochore assembly. The thermophilic budding yeast Kluyveromyces marxianus also has a point centromere but its length is nearly double the S. cerevisiae centromere and the number of centromeric nucleosomes and kinetochore attachment sites is unknown. Purification of native kinetochores from K. marxianus yielded a mixed population, with one subpopulation that appeared to consist of doublets, making it unclear whether K. marxianus shares the same attachment architecture as S. cerevisiae. Here, we demonstrate that though the doublet kinetochores have a functional impact on kinetochore strength, kinetochore localization throughout the cell cycle appears conserved between these two yeasts. In addition, whole spindle electron tomography demonstrates that a single microtubule binds to each chromosome. Single-molecule nucleosome mapping analysis suggests the presence of a single centromeric nucleosome. Taken together, we propose that the K. marxianus point centromere assembles a single centromeric nucleosome that mediates attachment to one microtubule.
]]></description>
<dc:creator>Barrero, D. J.</dc:creator>
<dc:creator>Hedouin, S.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Asbury, C. L.</dc:creator>
<dc:creator>Stergachis, A.</dc:creator>
<dc:creator>O'Toole, E.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2025-01-27</dc:date>
<dc:identifier>doi:10.1101/2025.01.24.634737</dc:identifier>
<dc:title><![CDATA[Centromeres in the thermotolerant yeast K. marxianusmediate attachment to a single microtubule]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.29.635299v1?rss=1">
<title>
<![CDATA[
Protein sequence editing defines distinct and overlapping functions of SKN-1A/Nrf1 and SKN-1C/Nrf2. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.29.635299v1?rss=1</link>
<description><![CDATA[
The Nrf/NFE2L family of transcription factors regulates redox balance, xenobiotic detoxification, metabolism, proteostasis, and aging. Nrf1/NFE2L1 is primarily responsible for stress-responsive upregulation of proteasome subunit genes and is essential for adaptation to proteotoxic stress. Nrf2/NFE2L2 is mainly involved in activating oxidative stress responses and promoting xenobiotic detoxification. Nrf1 and Nrf2 contain very similar DNA binding domains and can drive similar transcriptional responses. In C. elegans, a single gene, skn-1, encodes distinct protein isoforms, SKN-1A and SKN-1C, that function analogously to mammalian Nrf1 and Nrf2, respectively, and share an identical DNA binding domain. Thus, the extent to which SKN-1A/Nrf1 and SKN-1C/Nrf2 functions are distinct or overlapping has been unclear. Regulation of the proteasome by SKN-1A/Nrf1 requires post-translational conversion of N-glycosylated asparagine residues to aspartate by the PNG-1/NGLY1 peptide:N-glycanase, a process we term  sequence editing. Here, we reveal the consequences of sequence editing for the transcriptomic output of activated SKN-1A. We confirm that activation of proteasome subunit genes is strictly dependent on sequence editing. In addition, we find that sequence edited SKN-1A can also activate genes linked to redox homeostasis and xenobiotic detoxification that are also regulated by SKN-1C, but the extent of these genes activation is antagonized by sequence editing. Using mutant alleles that selectively inactivate either SKN-1A or SKN-1C, we show that both isoforms promote optimal oxidative stress resistance, acting as effectors for distinct signaling pathways. These findings suggest that sequence editing governs SKN-1/Nrf functions by tuning the SKN-1A/Nrf1 regulated transcriptome.
]]></description>
<dc:creator>Jochim, B. E.</dc:creator>
<dc:creator>Topalidou, I.</dc:creator>
<dc:creator>Lehrbach, N. J.</dc:creator>
<dc:date>2025-01-29</dc:date>
<dc:identifier>doi:10.1101/2025.01.29.635299</dc:identifier>
<dc:title><![CDATA[Protein sequence editing defines distinct and overlapping functions of SKN-1A/Nrf1 and SKN-1C/Nrf2.]]></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/10.1101/2025.02.05.636596v1?rss=1">
<title>
<![CDATA[
Modeling of hepatitis B virus infection spread in primary human hepatocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.05.636596v1?rss=1</link>
<description><![CDATA[
Chronic hepatitis B virus (HBV) infection poses a significant global health threat, causing severe liver diseases including cirrhosis and hepatocellular carcinoma. We characterized HBV DNA kinetics in primary human hepatocytes (PHH) over 32 days post-inoculation (pi) and used agent-based modeling (ABM) to gain insights into HBV lifecycle and spread. Parallel PHH cultures were mock-treated or HBV entry inhibitor Myr-preS1 (6.25 g/mL) was initiated 24h pi. In untreated PHH, 3 viral DNA kinetic patterns were identified: (1) an initial decline, followed by (2) rapid amplification, and (3) slower amplification/accumulation. In the presence of Myr-preS1, viral DNA and infected cell numbers in phase 3 were effectively blocked, with minimal to no increase. This suggests that phase 2 represents viral amplification in initially infected cells, while phase 3 corresponds to viral spread to naive cells. The ABM reproduced well the HBV kinetic patterns observed and predicted that the viral eclipse phase lasts between 18 and 38 hours. After the eclipse phase, the viral production rate increases over time, starting with a slow production cycle of 1 virion per day, which gradually accelerates to 1 virion per hour after 3 days. Approximately 4 days later, virion production reaches a steady state production rate of 4 virions/hour. The estimated median efficacy of Myr-preS1 in blocking HBV spread was 91% (range: 90-92%). The HBV kinetics and the predicted estimates of the HBV eclipse phase duration and HBV production cycles in PHH are similar of those predicted in uPA/SCID mice with human livers.

ImportanceWhile primary human hepatocytes (PHH) are the most physiologically relevant culture system for studying HBV infection in vitro, comprehensive understanding of HBV infection kinetics and spread in PHH is lacking. In this study, we characterize HBV viral kinetics and employ agent-based modeling (ABM) to provide quantitative insights into the HBV production cycle and viral spread in PHH. The ABM provides an estimate of HBV eclipse phase duration, HBV production cycles and Myr-preS1 efficacy in blocking HBV spread in PHH. The results resemble those predicted in uPA/SCID mice with human livers, demonstrating that estimated HBV infection kinetic parameters in PHH in vitro mirrors that observed in in vivo HBV infection chimeric mouse model.
]]></description>
<dc:creator>Shi, Z.</dc:creator>
<dc:creator>Tsuge, M.</dc:creator>
<dc:creator>Collier, N.</dc:creator>
<dc:creator>Takeuchi, Y.</dc:creator>
<dc:creator>Uchida, T.</dc:creator>
<dc:creator>Rutter, C.</dc:creator>
<dc:creator>Teraoka, Y.</dc:creator>
<dc:creator>Uprichard, S. L.</dc:creator>
<dc:creator>Ishida, Y.</dc:creator>
<dc:creator>Tateno, C.</dc:creator>
<dc:creator>Ozik, J.</dc:creator>
<dc:creator>Dahari, H.</dc:creator>
<dc:creator>Chayama, K.</dc:creator>
<dc:date>2025-02-07</dc:date>
<dc:identifier>doi:10.1101/2025.02.05.636596</dc:identifier>
<dc:title><![CDATA[Modeling of hepatitis B virus infection spread in primary human hepatocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.06.636949v1?rss=1">
<title>
<![CDATA[
Systems Pharmacology Screening Reveals Mertk and other Targetable Kinases in Supratentorial Ependymomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.06.636949v1?rss=1</link>
<description><![CDATA[
Ependymomas (EPN) are rare central nervous system tumors that account for approximately 10% of intracranial tumors in children and 4% in adults. Despite their clinical and molecular heterogeneity, spanning supratentorial, posterior fossa, and spinal subtypes, treatment remains limited to surgery and radiotherapy, with chemotherapy offering minimal benefit. Here, we performed transcriptomic analysis of 370 human ependymoma samples and identified two distinct molecular subgroups: EPN-E1 and EPN-E2. The EPN-E1 cluster is enriched for supratentorial tumors harboring ZFTA-RELA fusions (ZFTA-RELAfus), which occur in over 70% of cases and are associated with poor prognosis. To identify targeted therapies for this aggressive subtype, we validated a ZFTA-RELAfus mouse model that recapitulates the human EPN-E1 transcriptome and used it for target discovery. Through Kinome Regularization, a machine learning-driven polypharmacology approach, we identified MERTK as a critical regulator of tumor cell viability. Genetic depletion or pharmacologic inhibition of Mertk reduced cell growth ex vivo, and treatment with a clinical-grade MERTK inhibitor significantly suppressed tumor proliferation in vivo. Both human EPN-E1 tumors and ZFTA-RELAfus mouse tumors exhibited elevated expression of MERTK and its ligand GAS6, and MERTK inhibition led to suppression of pro-survival signaling pathways including MEK/ERK and PI3K/AKT. Notably, over 80% of genes upregulated in ZFTA-RELAfus tumors were downregulated following MERTK inhibition, indicating a strong dependency on this pathway for tumor maintenance. These findings define a signaling vulnerability in ZFTA-RELA-driven ependymomas and support the clinical development of MERTK-targeted therapies for patients with the high-risk EPN-E1 subtype.
]]></description>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Zhu, S.</dc:creator>
<dc:creator>Russell, Z.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Arakaki, A.</dc:creator>
<dc:creator>Kumasaka, D.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:creator>Michealraj, A.</dc:creator>
<dc:creator>Holland, E.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:date>2025-02-08</dc:date>
<dc:identifier>doi:10.1101/2025.02.06.636949</dc:identifier>
<dc:title><![CDATA[Systems Pharmacology Screening Reveals Mertk and other Targetable Kinases in Supratentorial Ependymomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.10.637482v1?rss=1">
<title>
<![CDATA[
Precise engineering of chimeric antigen receptor expression levels defines T cell identity and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.10.637482v1?rss=1</link>
<description><![CDATA[
Chimeric Antigen Receptor (CAR) T therapy is a potent treatment for haematological malignancies, but T cell exhaustion reduces its efficacy in many patients. Although high CAR transgene levels appear to drive T cell exhaustion, the relationship between CAR expression levels, T cell function, and transcriptional identity is yet to be mapped at high resolution. Here, we harness a high-resolution microRNA-based control system to precisely modulate CAR transgene expression levels and assess the impact on T cell activation, gene expression and function. By post-transcriptionally modulating CAR abundance, we show that differential CAR levels significantly impact T cell proliferation, cytokine production and tonic signalling. T cells with high CAR expression become strongly activated even at low target antigen densities, while those with low CAR expression are triggered only by high concentrations of their target. Single-cell RNA sequencing of primary T cells expressing a broad range of CAR transcript levels revealed global transcriptional programmes that become dysfunctional with increased CAR abundance, expanding our understanding of T cell exhaustion. Notably, we identified a narrow CAR expression range where the exhaustion transcriptional state is not triggered, demonstrating that T cell exhaustion can be controlled by fine-tuning CAR levels. This work demonstrates that CAR expression levels are key determinants of T cell transcriptional identity and function and introduces a tractable method to precisely tune CAR expression and T cell activity.
]]></description>
<dc:creator>Ramos, A. S.</dc:creator>
<dc:creator>Simon, S.</dc:creator>
<dc:creator>Siller-Farfan, J.</dc:creator>
<dc:creator>Rajan, A.</dc:creator>
<dc:creator>Revale, S.</dc:creator>
<dc:creator>Zanchini di Castiglionchio, E.</dc:creator>
<dc:creator>Pelea, O.</dc:creator>
<dc:creator>Fulga, T. A.</dc:creator>
<dc:creator>Riddell, S.</dc:creator>
<dc:creator>Dushek, O.</dc:creator>
<dc:creator>Michaels, Y. S.</dc:creator>
<dc:creator>Sauka-Spengler, T.</dc:creator>
<dc:date>2025-02-11</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.637482</dc:identifier>
<dc:title><![CDATA[Precise engineering of chimeric antigen receptor expression levels defines T cell identity and function]]></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/10.1101/2025.02.14.638232v1?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/10.1101/2025.02.12.637904v1?rss=1">
<title>
<![CDATA[
Defined cellular reprogramming of androgen receptor-active prostate cancer to neuroendocrine prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.12.637904v1?rss=1</link>
<description><![CDATA[
Neuroendocrine prostate cancer (NEPC) arises primarily through neuroendocrine transdifferentiation (NEtD) as an adaptive mechanism of therapeutic resistance. Models to define the functional effects of putative drivers of this process on androgen receptor (AR) signaling and NE cancer lineage programs are lacking. We adapted a genetically defined strategy from the field of cellular reprogramming to directly convert AR-active prostate cancer (ARPC) to AR-independent NEPC using candidate factors. We delineated critical roles of the pioneer factors ASCL1 and NeuroD1 in NEtD and uncovered their abilities to silence AR expression and signaling by remodeling chromatin at the somatically acquired AR enhancer and global AR binding sites with enhancer activity. We also elucidated the dynamic temporal changes in the transcriptomic and epigenomic landscapes of cells undergoing acute lineage conversion from ARPC to NEPC which should inform future therapeutic development. Further, we distinguished the activities of ASCL1 and NeuroD1 from the inactivation of RE-1 silencing transcription factor (REST), a master suppressor of a major neuronal gene program, in establishing a NEPC lineage state and in modulating the expression of genes associated with major histocompatibility complex class I (MHC I) antigen processing and presentation. These findings provide important, clinically relevant insights into the biological processes driving NEtD of prostate cancer.
]]></description>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Song, K.</dc:creator>
<dc:creator>Sun, H.</dc:creator>
<dc:creator>Tao, Y.</dc:creator>
<dc:creator>Huang, A.</dc:creator>
<dc:creator>Bhatia, V.</dc:creator>
<dc:creator>Hanratty, B.</dc:creator>
<dc:creator>Patel, R. A.</dc:creator>
<dc:creator>Long, H. W.</dc:creator>
<dc:creator>Morrissey, C.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Graeber, T. G.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:date>2025-02-17</dc:date>
<dc:identifier>doi:10.1101/2025.02.12.637904</dc:identifier>
<dc:title><![CDATA[Defined cellular reprogramming of androgen receptor-active prostate cancer to neuroendocrine prostate 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/10.1101/2025.02.12.637975v1?rss=1">
<title>
<![CDATA[
Defined media reveals the essential role of lipid scavenging to support cancer cell proliferation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.12.637975v1?rss=1</link>
<description><![CDATA[
Fetal bovine serum (FBS) is a nearly ubiquitous, yet undefined additive in mammalian cell culture media whose functional contributions to promoting cell proliferation remain poorly understood. Efforts to replace serum supplementation in culture media have been hindered by an incomplete understanding of the environmental requirements fulfilled by FBS in culture. Here, we use a combination of live-cell imaging and liquid chromatography-mass spectrometry to elucidate the role of serum in supporting proliferation. We show that serum provides consumed factors that enable proliferation and demonstrate that the serum metal and lipid components are crucial to sustaining proliferation in culture. Importantly, despite access to a wide range of lipid classes, albumin-bound lipids are the primary species consumed during cancer cell proliferation. Furthermore, we find that combinations of the additive ITS, containing necessary metals, and albumin-associated lipid classes are sufficient to replace FBS in culture media. We show that serum-free media enables sensitive quantification of lipid consumption dynamics during cell proliferation, which indicate that fatty acids (FA) are consumed through a mass-action mechanism, with minimal competition from other lipid classes. Finally, we find that pharmacologic disruption of FA activation and incorporation into the cellular lipidome reduces uptake from the environment and impairs cell proliferation. This work therefore identifies metabolic contributions of serum in cell culture settings and provides a framework for building cell culture systems that sustain cell proliferation without the variable and undefined contributions of FBS.
]]></description>
<dc:creator>Newsom, O. J.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:date>2025-02-17</dc:date>
<dc:identifier>doi:10.1101/2025.02.12.637975</dc:identifier>
<dc:title><![CDATA[Defined media reveals the essential role of lipid scavenging to support cancer cell proliferation]]></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/10.1101/2025.02.13.638022v1?rss=1">
<title>
<![CDATA[
Accuracy and Scalability of Machine Learning Methods for Genotype-Phenotype Association Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.13.638022v1?rss=1</link>
<description><![CDATA[
Many machine learning methods can be applied to predicting phenotypes from genetic data. Which of these methods work best remains an open question, however. To answer this question, we propose to compare a variety of approaches ability to predict a simulated non-linear complex trait. Specifically, we evaluate these methods on their accuracy and scalability with respect to the amount training data available, the noise present in the data, the complexity of the simulated (trait) functions, and their ability to provide insight into the simulated trait. We then compare the best approach to state-of-the-art models in real data, predicting gout in the UK Biobank. We find that transformer encoders outperform all other methods in simulations, and perform comparably to the state-of-the-art with real data, with a promise to scale to significantly larger datasets.
]]></description>
<dc:creator>Collienne, K.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Gavryushkin, A.</dc:creator>
<dc:date>2025-02-17</dc:date>
<dc:identifier>doi:10.1101/2025.02.13.638022</dc:identifier>
<dc:title><![CDATA[Accuracy and Scalability of Machine Learning Methods for Genotype-Phenotype Association Data]]></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/10.1101/2025.02.18.638948v1?rss=1">
<title>
<![CDATA[
Succinate Dehydrogenase loss causes cascading metabolic effects that impair pyrimidine biosynthesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.18.638948v1?rss=1</link>
<description><![CDATA[
Decreased availability of the amino acid aspartate can constrain cell function in diverse biological contexts, but the temporal interplay between aspartate, downstream metabolic changes, and functional effects remains poorly understood. Using an aspartate biosensor and live-cell imaging, we examine the interaction between aspartate abundance and cell proliferation in several models of aspartate limitation. While aspartate deficiencies intuitively interface with proliferation in some contexts, aspartate limitation from succinate dehydrogenase (SDH) inhibition causes strikingly nonintuitive dynamics resulting from an outsized impairment of pyrimidine synthesis. Mechanistically, we find that SDH loss impairs pyrimidine biosynthesis by decreasing aspartate and accumulating succinate, which competitively inhibits mammalian aspartate transcarbamylase (ATCase). This metabolic interaction persists in multiple models of SDH deficiency, causing pyrimidine insufficiency, replication stress, and sensitivity to ATR kinase inhibition. These findings define a novel role for succinate in modulating cellular nucleotide homeostasis, suggest a potential therapeutic vulnerability of SDH-deficient tumors, and demonstrate how cascading metabolic interactions can unfold to impact cell function.
]]></description>
<dc:creator>Hart, M. L.</dc:creator>
<dc:creator>Davidsen, K.</dc:creator>
<dc:creator>Danquah, S.</dc:creator>
<dc:creator>Zheng, E.</dc:creator>
<dc:creator>Sokolov, D.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:date>2025-02-19</dc:date>
<dc:identifier>doi:10.1101/2025.02.18.638948</dc:identifier>
<dc:title><![CDATA[Succinate Dehydrogenase loss causes cascading metabolic effects that impair pyrimidine biosynthesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.19.636902v1?rss=1">
<title>
<![CDATA[
A Syntenic Pangenome for Gardnerella Reveals Taxonomic Boundaries and Stratification of Metabolic and Virulence Potential across Species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.19.636902v1?rss=1</link>
<description><![CDATA[
Gardnerella species are central to bacterial vaginosis (BV), a condition affecting nearly one in three women of reproductive age and associated with preterm birth and increased susceptibility to sexually transmitted infections. Despite decades of study, progress in defining Gardnerella diversity has been hindered by inconsistent taxonomy and poor-quality genomic resources. Here, we sequenced 392 new Gardnerella isolates from asymptomatic and BV-associated microbiota and integrated this collection with all publicly available genomes. After stringent curation, we generated a high-quality reference set of 313 distinct genomes that underpins a comprehensive taxonomic framework. Using average nucleotide identity (ANI), digital DNA-DNA hybridization (dDDH), and phylogenomics, we resolved 21 genomic lineages encompassing 11 species and 15 subspecies, each assigned a provisional formal name. Integration of complete long-read assemblies enabled construction of the first syntenic Gardnerella pangenome, revealing lineage-specific repertoires of virulence, metabolic, and defense systems - including variable sialidases (NanH), vaginolysin, and amino-acid biosynthetic pathways - and defining conserved genomic architecture across species. Comparative methylome profiling further highlighted restriction-modification system diversity that may influence genetic exchange. Finally, we identified the first native cryptic plasmids in Gardnerella, overturning the assumption that the genus lacks plasmids, and demonstrated their use in generating a replicative E. coli-Gardnerella shuttle vector. Together, these results establish a complete genomic and functional framework for Gardnerella, providing a reproducible foundation for mechanistic and translational studies of BV and a model for resolving taxonomy and functional stratification in other urogenital-associated bacteria.
]]></description>
<dc:creator>Bouzek, H.</dc:creator>
<dc:creator>Srinivasan, S.</dc:creator>
<dc:creator>Jones, D. S.</dc:creator>
<dc:creator>McMahon, E. F.</dc:creator>
<dc:creator>Strenk, S. M.</dc:creator>
<dc:creator>Fiedler, T. L.</dc:creator>
<dc:creator>Fredricks, D. N.</dc:creator>
<dc:creator>Johnston, C. D.</dc:creator>
<dc:date>2025-02-19</dc:date>
<dc:identifier>doi:10.1101/2025.02.19.636902</dc:identifier>
<dc:title><![CDATA[A Syntenic Pangenome for Gardnerella Reveals Taxonomic Boundaries and Stratification of Metabolic and Virulence Potential across Species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.19.639046v1?rss=1">
<title>
<![CDATA[
Multi-scale machine learning model predicts muscle and functional disease progression in FSHD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.19.639046v1?rss=1</link>
<description><![CDATA[
Facioscapulohumeral muscular dystrophy (FSHD) is a genetic neuromuscular disorder characterized by progressive muscle degeneration with substantial variability in severity and progression patterns. FSHD is a highly heterogeneous disease; however, current clinical metrics used tracking disease progression lack sensitivity for personalized assessment, which greatly limits the design and execution of clinical trials. This study introduces a multi-scale machine learning framework leveraging whole-body magnetic resonance imaging (MRI) and clinical data to predict regional, muscle, joint, and functional progression in FSHD. The goal this work is to create a  digital twin of individual FSHD patients that can be leveraged in clinical trials.

Using a combined dataset of over 100 patients from seven studies, MRI-derived metrics--including fat fraction, lean muscle volume, and fat spatial heterogeneity at baseline--were integrated with clinical and functional measures. A three-stage random forest model was developed to predict annualized changes in muscle composition and a functional outcome (timed up-and-go (TUG)). All model stages revealed strong predictive performance in separate holdout datasets. After training, the models predicted fat fraction change with a root mean square error (RMSE) of 2.16% and lean volume change with a RMSE of 8.1ml in a holdout testing dataset. Feature analysis revealed that metrics fat heterogeneity within muscle predicts muscle-level progression. The stage 3 model that combined functional muscle groups and predicted change in TUG with a RMSE of 0.6 seconds, in the holdout testing dataset. This study demonstrates the machine learning models incorporating individual muscle and performance data can effectively predict MRI disease progression and functional performance of complex tasks, addressing the heterogeneity and nonlinearity inherent in FSHD. Further studies incorporating larger longitudinal cohorts as well as comprehensive clinical and functional measures will allow for expanding and refining this model. As many neuromuscular diseases are characterized by varability and heterogeneity similar to FSHD, such approaches have broad applicability.
]]></description>
<dc:creator>Blemker, S. S.</dc:creator>
<dc:creator>Riem, L.</dc:creator>
<dc:creator>DuCharme, O.</dc:creator>
<dc:creator>Pinette, M.</dc:creator>
<dc:creator>Costanzo, K. E.</dc:creator>
<dc:creator>Weatherley, E.</dc:creator>
<dc:creator>Statland, J.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:creator>Wang, L. H.</dc:creator>
<dc:creator>Shaw, D. W. W.</dc:creator>
<dc:creator>The Move+ Investigator Team,</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Leung, D.</dc:creator>
<dc:creator>Friedman, S.</dc:creator>
<dc:date>2025-02-20</dc:date>
<dc:identifier>doi:10.1101/2025.02.19.639046</dc:identifier>
<dc:title><![CDATA[Multi-scale machine learning model predicts muscle and functional disease progression in FSHD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.19.639175v1?rss=1">
<title>
<![CDATA[
A discrete region of the D4Z4 is sufficient to initiate epigenetic silencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.19.639175v1?rss=1</link>
<description><![CDATA[
The DUX4 transcription factor is briefly expressed in the early embryo and is epigenetically repressed in somatic tissues. Loss of epigenetic repression can result in the aberrant expression of DUX4 in skeletal muscle and can cause facioscapulohumeral dystrophy (FSHD). Multiple factors have been identified as necessary to maintain epigenetic silencing of DUX4 in skeletal muscle, but whether specific sequences at the DUX4 locus are sufficient for epigenetic silencing has been unknown. We cloned fragments of the D4Z4 macrosatellite repeat, the DNA region that encompasses the DUX4 retrogene, adjacent to a reporter driven by a constitutive promoter and identified a single fragment sufficient to epigenetically repress reporter gene expression. Previously identified suppressors of DUX4 expression--SETDB1, ATF7IP, SIN3A/B, and LRIF1--were necessary for silencing activity and p38 inhibitors enhanced suppression. These findings identify a key regulatory sequence for D4Z4 epigenetic repression and establish a model system for mechanistic and discovery studies.
]]></description>
<dc:creator>Paatela, E. M.</dc:creator>
<dc:creator>St. Amant, F. G.</dc:creator>
<dc:creator>Hamm, D. C.</dc:creator>
<dc:creator>Bennett, S. R.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:creator>van der Maarel, S. M.</dc:creator>
<dc:creator>Tapscott, S. J.</dc:creator>
<dc:date>2025-02-23</dc:date>
<dc:identifier>doi:10.1101/2025.02.19.639175</dc:identifier>
<dc:title><![CDATA[A discrete region of the D4Z4 is sufficient to initiate epigenetic silencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.05.640197v1?rss=1">
<title>
<![CDATA[
Pooled CAR-T screening in nonhuman primates identifies designs with enhanced proliferation, trafficking, and persistence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.05.640197v1?rss=1</link>
<description><![CDATA[
Chimeric antigen receptor T (CAR-T) cell therapy has revolutionized treatment for B-cell malignancies, yet over 60% of patients relapse within one year, often due to insufficient CAR-T persistence. While mouse and primary cell models have been instrumental in advancing CAR-T therapy, they frequently fail to predict clinical outcomes, underscoring the need for more translationally relevant models. To address this limitation, we conducted the first systematic evaluation of CAR structure-function relationships in an immunocompetent nonhuman primate (NHP) model. We engineered an array of 20 CD20-targeted CARs with distinct combinations of hinge, transmembrane, and costimulatory domains. Following ex vivo characterization, we administered pooled autologous CAR-T arrays to three NHPs and tracked CAR abundance longitudinally using a novel digital droplet PCR assay. Ex vivo, CAR-T cells incorporating the MyD88-CD40 costimulatory domain exhibited markedly distinct functional profiles, including increased activation, unique cytokine secretion, tonic signaling, and resistance to exhaustion. In vivo, MyD88-CD40 CARs expanded dramatically, comprising up to 100% of peripheral T cells and significantly outperforming canonical CD28- and 4-1BB-based CARs. This expansion was associated with robust B-cell depletion across all animals. MyD88-CD40 CARs, particularly those with a CD28 hinge and transmembrane domain, demonstrated superior trafficking to secondary lymphoid tissues and persistence through study endpoint, unlike other CARs which waned by day 28. Our findings highlight the value of NHP models for screening CAR designs and identify MyD88-CD40 CARs as candidates with unmatched potency. The unique functional attributes conferred by this domain may provide key insights into features that drive enhanced CAR-T cell activity.

Key pointsO_LIWe developed the first pooled CAR-T screening platform in an immunocompetent nonhuman primate model to directly compare CAR designs.
C_LIO_LIWe identified MyD88-CD40 costimulatory domain as vastly superior to conventional domains in proliferation, trafficking and persistence.
C_LI
]]></description>
<dc:creator>Maynard, L. H.</dc:creator>
<dc:creator>Cavanaugh, E. J.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Starke, C. E.</dc:creator>
<dc:creator>Doherty, S. M.</dc:creator>
<dc:creator>Einhaus, T.</dc:creator>
<dc:creator>Perez, A.</dc:creator>
<dc:creator>Stensland, L.</dc:creator>
<dc:creator>Hoffman, M.</dc:creator>
<dc:creator>Nelson, V.</dc:creator>
<dc:creator>Herrin, S.</dc:creator>
<dc:creator>Littlewood, C.</dc:creator>
<dc:creator>Camou, K.</dc:creator>
<dc:creator>Wilson, E.</dc:creator>
<dc:creator>Wessel, C.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Kiem, H.-P.</dc:creator>
<dc:creator>Peterson, C. W.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.05.640197</dc:identifier>
<dc:title><![CDATA[Pooled CAR-T screening in nonhuman primates identifies designs with enhanced proliferation, trafficking, and persistence]]></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/10.1101/2025.03.09.642243v1?rss=1">
<title>
<![CDATA[
Total whole-arm chromosome losses predict malignancy in human cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.09.642243v1?rss=1</link>
<description><![CDATA[
Aneuploidy is observed as gains or losses of whole chromosomes or chromosome arms and is a common hallmark of cancer. Whereas models for the generation of aneuploidy in cancer invoke mitotic chromosome segregation errors, whole-arm losses might occur simply as a result of centromere breakage. We recently showed that elevated RNA Polymerase II (RNAPII) level over S-phase-dependent histone genes predicts rapid recurrence of human meningioma and is correlated with total whole-arm losses relative to gains. To explain this imbalance in arm losses over gains, we have proposed that histone overexpression at S-phase competes with the histone H3 variant CENP-A, resulting in centromere breaks and whole-arm losses. To test whether centromere breaks alone can drive aneuploidy, we ask whether total whole-arm aneuploids can predict outcome across different cancer types in large RNA and whole-genome sequencing databanks. We find that total whole-arm losses generally predict outcome, suggesting that centromere breakage is a major initiating factor leading to aneuploidy and the resulting changes in the selective landscape that drive most cancers. We also present evidence that centromere breakage alone is sufficient to account for whole-arm losses and gains, contrary to mitotic spindle error models for generation of aneuploidy. Our results suggest that therapeutic intervention targeting histone overexpression has the potential of reducing aneuploidy and slowing cancer progression.

Significance StatementGain or loss of whole chromosome arms following centromere breaks is frequent in cancer, but whether or not there is a common initiating event is unknown. Here we show that the total number of whole-arm losses predicts patient outcomes across cancer types, suggesting a causal relationship. This general excess of losses over gains is not predicted by mitotic error models of aneuploidy but rather suggests that centromere breaks themselves initiate whole-arm aneuploidies. Insofar as aneuploidy reshapes the selective landscapes that drive most cancers, our results have potential clinical implications.
]]></description>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2025-03-11</dc:date>
<dc:identifier>doi:10.1101/2025.03.09.642243</dc:identifier>
<dc:title><![CDATA[Total whole-arm chromosome losses predict malignancy in human cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.12.642815v1?rss=1">
<title>
<![CDATA[
Delta-catenin is required for cell proliferation in virus positive Merkel cell carcinoma cell lines but not in human fibroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.12.642815v1?rss=1</link>
<description><![CDATA[
Merkel cell carcinoma (MCC) is a highly aggressive neuroendocrine skin cancer often driven by the integration of Merkel cell polyomavirus (MCPyV) into the host genome and the persistent expression of its viral oncoproteins, small tumor (ST) antigen and truncated large tumor (t-LT) antigen. While human fibroblasts support MCPyV replication, the cell of origin for MCC remains unknown. We hypothesized that MCPyV initially replicates in fibroblasts but, in rare cases, infects Merkel cell progenitors, contributing to MCC development. Using TurboID mass spectrometry, we identified {delta}-catenin as a novel ST interactor in fibroblasts. However, while ST binds {delta}-catenin in fibroblasts, this interaction is absent in virus-positive (VP)-MCC cell lines. Despite this, {delta}-catenin is essential for VP-MCC, but not for fibroblast, cell proliferation. We found that fibroblasts predominantly express {delta}-catenin isoform 1, whereas VP-MCC cells mainly express isoform 3. Overexpression of isoform 1 in VP-MCC failed to restore ST binding. {delta}-catenin promotes VP-MCC proliferation by regulating cell cycle gene expression through its interaction with Kaiso, a transcriptional repressor. Additionally, we found that LSD1 (KDM1A) regulates {delta}-catenin isoform 3 expression by modulating ESRP1, a {delta}-catenin splicing factor. Our findings reveal novel host factors involved in MCPyV infection and MCC tumorigenesis, suggesting that the host cell supporting viral replication and the MCC cell of origin may be distinct cell types.

ImportanceMerkel cell polyomavirus (MCPyV), the only known human oncogenic polyomavirus, is the primary cause of Merkel cell carcinoma (MCC), a rare and aggressive type of skin cancer. MCC is driven by two viral proteins: small T (ST) and large T (LT). While the virus can replicate in skin fibroblasts, it is still unknown which type of skin cell becomes cancerous. We found that ST binds to a host protein, {delta}-catenin in fibroblasts, potentially playing a role in the virus lifecycle, but this interaction is missing in the cancer cells. Our study provides evidence that the cells in which the virus replicates and causes cancer are different.
]]></description>
<dc:creator>Landazuri Vinueza, J.</dc:creator>
<dc:creator>Salisbury, N. J. H.</dc:creator>
<dc:creator>Dye, K.</dc:creator>
<dc:creator>Roman, A.</dc:creator>
<dc:creator>Galloway, D. A.</dc:creator>
<dc:date>2025-03-14</dc:date>
<dc:identifier>doi:10.1101/2025.03.12.642815</dc:identifier>
<dc:title><![CDATA[Delta-catenin is required for cell proliferation in virus positive Merkel cell carcinoma cell lines but not in human fibroblasts]]></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/10.1101/2025.03.14.643253v1?rss=1">
<title>
<![CDATA[
Capsule-Based Single-Cell Genome Sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.14.643253v1?rss=1</link>
<description><![CDATA[
Single-cell genomics methods have unveiled the heterogeneity present in seemingly homogenous populations of cells, however, these techniques require meticulous optimization. How exactly does one handle and manipulate the biological contents from a single cell? Here, we introduce and characterize a novel semi-permeable capsule (SPC), capable of isolating single cells and their contents while facilitating biomolecular exchange based on size-selectivity. These capsules maintain stability under diverse physical and chemical conditions and allow selective diffusion of biomolecules, effectively retaining larger biomolecules including genomic DNA, and cellular complexes, while permitting the exchange of smaller molecules, including primers and enzymes. We demonstrate the utility of SPCs for single cell assays by performing the simultaneous culture of over 500,000 cellular colonies, demonstrating efficient and unbiased nucleic acid amplification, and performing combinatorial indexing-based single-cell whole genome sequencing (sc-WGS). Notably, SPC-based sc-WGS facilitates uniform genome coverage and minimal cross-contamination allowing for the detection of genomic variants with high sensitivity and specificity. Leveraging these properties, we conducted a proof-of-concept lineage tracing experiment using cells harboring the hypermutator polymerase {varepsilon} allele (POLE P286R). Sequencing of 1000 single cell genomes at low depth facilitated the capture of lineage marks deposited throughout the genome during each cell division and the subsequent reconstruction of cellular genealogies. Capsule-based sc-WGS expands the single-cell genomics toolkit and will facilitate the investigation of somatic variants, resolved to single cells at scale.
]]></description>
<dc:creator>Mullaney, D. B.</dc:creator>
<dc:creator>Sgrizzi, S. R.</dc:creator>
<dc:creator>Mai, D.</dc:creator>
<dc:creator>Campbell, I.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Sinkunas, A.</dc:creator>
<dc:creator>Kerr, D. L.</dc:creator>
<dc:creator>Browning, V. E.</dc:creator>
<dc:creator>Eisenach, H. E.</dc:creator>
<dc:creator>Sims, J. N.</dc:creator>
<dc:creator>Nichols, E. K.</dc:creator>
<dc:creator>Lapointe, C. P.</dc:creator>
<dc:creator>Amimura, Y.</dc:creator>
<dc:creator>Harris, K.</dc:creator>
<dc:creator>Zilionis, R.</dc:creator>
<dc:creator>Srivatsan, S. R.</dc:creator>
<dc:date>2025-03-17</dc:date>
<dc:identifier>doi:10.1101/2025.03.14.643253</dc:identifier>
<dc:title><![CDATA[Capsule-Based Single-Cell Genome Sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.19.644145v1?rss=1">
<title>
<![CDATA[
Collagen-binding IL-12 expressing STEAP1 CAR-T cells reduce toxicity and eradicate mouse prostate cancer in combination with checkpoint inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.19.644145v1?rss=1</link>
<description><![CDATA[
Immunosuppressive microenvironments, the lack of immune infiltration, and antigen heterogeneity pose significant challenges for chimeric antigen receptor (CAR)-T cell therapies to tackle solid tumors. CAR-T cells were armed with immunostimulatory payloads, such as interleukin-12 (IL-12), to overcome this issue, but faced intolerable toxicity during clinical development. Here, we show that collagen-binding domain-fused IL-12 (CBD-IL-12) was retained within syngeneic murine prostate tumors, after secretion from CAR-T cells targeting human six transmembrane epithelial antigen of the prostate 1 (STEAP1). This led to equivalently high intratumoral interferon-{gamma} levels without hepatotoxicity and infiltration of T cells into non-target organs, compared with unmodified IL-12. Both innate and adaptive immune compartments were dramatically activated and recognized diverse tumor antigens after CBD-IL-12 CAR-T cell treatment. Combination immunotherapy of CBD-IL-12 CAR-T cells and immune checkpoint inhibitors eradicated large tumors in an established prostate cancer model, without pre-conditioning chemotherapy. The therapy generated anti-tumor immunological memory. CBD-fusion to potent yet toxic payloads of CAR-T therapy may remove obstacles to their clinical translation towards elimination of solid tumors.
]]></description>
<dc:creator>Sasaki, K.</dc:creator>
<dc:creator>Bhatia, V.</dc:creator>
<dc:creator>Asano, Y.</dc:creator>
<dc:creator>Bakhtiari, J.</dc:creator>
<dc:creator>Kaur, P.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Matsuo, T.</dc:creator>
<dc:creator>Nikolaidi, M.</dc:creator>
<dc:creator>Chiu, P.-C.</dc:creator>
<dc:creator>Dubois, O.</dc:creator>
<dc:creator>Chong, T.</dc:creator>
<dc:creator>Javier, G.</dc:creator>
<dc:creator>Priceman, S. J.</dc:creator>
<dc:creator>Chapuis, A. G.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:creator>Ishihara, J.</dc:creator>
<dc:date>2025-03-19</dc:date>
<dc:identifier>doi:10.1101/2025.03.19.644145</dc:identifier>
<dc:title><![CDATA[Collagen-binding IL-12 expressing STEAP1 CAR-T cells reduce toxicity and eradicate mouse prostate cancer in combination with checkpoint inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.26.645558v1?rss=1">
<title>
<![CDATA[
PglZ from Type I BREX phage defence systems is a metal-dependent nuclease that forms a sub-complex with BrxB 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.26.645558v1?rss=1</link>
<description><![CDATA[
BREX (Bacteriophage Exclusion) systems, identified through shared identity with Pgl (Phage Growth Limitation) systems, are a widespread, highly diverse group of phage defence systems found throughout bacteria and archaea. The varied BREX Types harbour multiple protein subunits (between four and eight) and all encode a conserved putative phosphatase (PglZ aka BrxZ) and an equally conserved, putative ATPase (BrxC). Almost all BREX systems also contain a site-specific methyltransferase (PglX aka BrxX). Despite having determined the structure and fundamental biophysical and biochemical behaviours for the PglX methyltransferase, the BrxL effector, the BrxA DNA-binding protein and the BrxR transcriptional regulator, the mechanism by which BREX impedes phage replication remains largely undetermined. In this study, we identify a stable BREX sub-complex of PglZ:BrxB, validate the structure and dynamic behaviour of that sub-complex, and assess the biochemical activity of PglZ, revealing it to be a metal-dependent nuclease. PglZ can cleave cyclic oligonucleotides, linear oligonucleotides, plasmid DNA and both non-modified and modified linear phage genomes. PglZ nuclease activity has no obvious role in BREX-dependent methylation, but does contribute to BREX phage defence. BrxB binding does not impact PglZ nuclease activity. These data contribute to our growing understanding of the BREX phage defence mechanism.
]]></description>
<dc:creator>Readshaw, J.</dc:creator>
<dc:creator>Doyle, L.</dc:creator>
<dc:creator>Puiu, M.</dc:creator>
<dc:creator>Kelly, A.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Kaiser, A.</dc:creator>
<dc:creator>McGuire, S.</dc:creator>
<dc:creator>Peralata-Acosta, J.</dc:creator>
<dc:creator>Smith, D.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>Kaiser, B. K.</dc:creator>
<dc:creator>Blower, T. R.</dc:creator>
<dc:date>2025-03-26</dc:date>
<dc:identifier>doi:10.1101/2025.03.26.645558</dc:identifier>
<dc:title><![CDATA[PglZ from Type I BREX phage defence systems is a metal-dependent nuclease that forms a sub-complex with BrxB]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.24.644993v1?rss=1">
<title>
<![CDATA[
TP53 Inactivation Confers Resistance to the Menin Inhibitor Revumenib in Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.24.644993v1?rss=1</link>
<description><![CDATA[
Acute myeloid leukemia (AML) is a heterogeneous cancer that is associated with poor outcomes. Revumenib and other menin inhibitors have shown promising activity against AMLs with KMT2A-rearrangements or NPM1 mutations. However, mechanisms of de novo resistance have not yet been elucidated. We analyzed a panel of cell lines and generated an isogenic model to assess the impact of TP53 mutations on the response of AML cells to revumenib. TP53 mutations are associated with de novo resistance to revumenib, impaired induction of TP53 transcriptional targets, and deregulated expression of the BH3 proteins BCL-2 and MCL-1. The MCL-1 inhibitor MIK665, but not venetoclax, preferentially sensitized TP53-mutant AML cells to revumenib. These data identify mutant TP53 as a potential biomarker for de novo resistance to revumenib, and provide a rationale to evaluate MCL-1 and menin inhibitor combinations in patients KMT2A-rearranged leukemias with TP53 mutations.
]]></description>
<dc:creator>D'Souza, J.</dc:creator>
<dc:creator>Leung, C. J.</dc:creator>
<dc:creator>Ballapuram, A. C.</dc:creator>
<dc:creator>Lin, A. S.</dc:creator>
<dc:creator>Batingana, A. R.</dc:creator>
<dc:creator>Lamble, A. J.</dc:creator>
<dc:creator>Ries, R. E.</dc:creator>
<dc:creator>Morales, C. E.</dc:creator>
<dc:creator>Cottonham, C.</dc:creator>
<dc:creator>Gona, P.</dc:creator>
<dc:creator>Fan, L.</dc:creator>
<dc:creator>Ma, X.</dc:creator>
<dc:creator>Shannon, K.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Braun, B. S.</dc:creator>
<dc:creator>Huang, B. J.</dc:creator>
<dc:date>2025-03-27</dc:date>
<dc:identifier>doi:10.1101/2025.03.24.644993</dc:identifier>
<dc:title><![CDATA[TP53 Inactivation Confers Resistance to the Menin Inhibitor Revumenib in Acute Myeloid Leukemia]]></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/10.1101/2025.04.04.645037v1?rss=1">
<title>
<![CDATA[
The bone marrow niche and hematopoietic system are distinctly remodeled by CD45-targeted astatine-211 radioimmunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.04.645037v1?rss=1</link>
<description><![CDATA[
Radioimmunotherapy (RIT) is used to treat patients with hematological malignancies known to infiltrate the bone marrow (BM) microenvironment. RIT uses target-specific monoclonal antibodies stably conjugated to radionuclides to deliver cytotoxic radiation to cells of interest. While RIT is effective at delivering radiation to cancer cells, normal tissue is also exposed to radiation upon RIT, the consequences of which are largely unknown. Here, we studied the cellular and molecular effects of CD45-targeted astatine-211 (211At) RIT, IgG non-targeted 211At RIT, and Cesium-137 total-body irradiation (TBI) on hematopoietic cells and their BM niche in wild-type immunocompetent mice. Relative to non-targeted RIT or TBI, CD45-targeted RIT significantly delayed hematopoietic regeneration overall in the peripheral blood and BM and reduced hematopoietic stem/progenitor cell recovery and colony-forming ability. While BM endothelial cells (ECs) do not express the CD45 antigen, CD45-targeted RIT significantly depleted BM ECs compared to non-targeted RIT or TBI. RNA sequence analysis revealed significantly different transcriptomic profiles of BM ECs from CD45-RIT-treated mice compared to non- targeted RIT or TBI. ECs from CD45-RIT-treated mice, but not TBI or IgG-RIT-treated mice, were transcriptionally enriched for TGF{beta}, NOTCH, and IFN signaling pathways compared to untreated mice. Collectively, our study indicates that CD45-targeted RIT severely impacts hematopoietic and EC niche recovery compared to non- targeted approaches. Future studies are required to determine the long-term consequences of such RIT-driven effects on BM niche physiology and how BM niche reprogramming by RIT affects cancer cells.

KEY POINTSO_LICD45-targeted radioimmunotherapy more effectively suppresses the hematopoietic system than non- targeted radiation delivery.
C_LIO_LIThe bone marrow vascular niche is differentially reprogrammed by CD45-targeted radioimmunotherapy compared to non-targeted radiation delivery.
C_LI
]]></description>
<dc:creator>Hagen, M. W.</dc:creator>
<dc:creator>Setiawan, N. J.</dc:creator>
<dc:creator>Dexter, S. L.</dc:creator>
<dc:creator>Woodruff, K. A.</dc:creator>
<dc:creator>Gaerlan, F. K.</dc:creator>
<dc:creator>Orozco, J. J.</dc:creator>
<dc:creator>Termini, C. M.</dc:creator>
<dc:date>2025-04-05</dc:date>
<dc:identifier>doi:10.1101/2025.04.04.645037</dc:identifier>
<dc:title><![CDATA[The bone marrow niche and hematopoietic system are distinctly remodeled by CD45-targeted astatine-211 radioimmunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.02.645322v1?rss=1">
<title>
<![CDATA[
Implementation of an adaptive-optics assisted isoSTED nanoscope 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.02.645322v1?rss=1</link>
<description><![CDATA[
The highest level of three-dimensional (3D) resolution in stimulated emission depletion (STED) nanoscopy involves harnessing a 4Pi architecture using two opposing objectives. This protocol describes the construction and alignment of a 4Pi-STED nanoscope, commonly referred to as an isoSTED nanoscope. It guides interested researchers through assembling optomechanical components, configuring electronic and control devices, aligning the optical beam path, and assessing the instruments performance. Designed for adept optical instrument builders, this protocol offers a detailed roadmap for constructing an isoSTED nanoscope with adaptive optics (AO) in approximately 12 months. With this finely calibrated instrument, researchers can achieve 3D biological images with isotropic sub-50-nm resolution in thick samples up to 35 {micro}m in depth.
]]></description>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Allgeyer, E. S.</dc:creator>
<dc:creator>Schroeder, L. K.</dc:creator>
<dc:creator>Tu, S.</dc:creator>
<dc:creator>Bewersdorf, J.</dc:creator>
<dc:creator>Hao, X.</dc:creator>
<dc:date>2025-04-06</dc:date>
<dc:identifier>doi:10.1101/2025.04.02.645322</dc:identifier>
<dc:title><![CDATA[Implementation of an adaptive-optics assisted isoSTED nanoscope]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.31.646278v1?rss=1">
<title>
<![CDATA[
HPV16 neutralizing monoclonal antibodies show evidence for common developmental pathways and public epitopes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.31.646278v1?rss=1</link>
<description><![CDATA[
Antibodies to human papillomavirus (HPV) primarily recognize surface exposed residues on five loops of the major capsid protein (L1) that vary significantly among HPV types. We determined which loops were required for neutralization for 70 HPV16 specific human monoclonal antibodies (mAbs) cloned from participants who received an HPV vaccine, and we describe molecular features of those antibodies.

Chimeric HPV16 pseudovirus (cpsV), each having one surface loop bearing multiple amino acid substitutions, were used to determine neutralization specificity. The HPV16-FG-loop was the loop most frequently required for neutralization (44 of 70, 62.9%), however, all other surface loops were used for neutralization by multiple mAbs: HI (13, 18.6%), DE (15, 21.4%), EF (six, 8.6%), BC (four, 5.7%). Antibodies that required multiple loops were common (17, 24.3%). Three mAbs (4.3%) required sequences on the c-terminus of L1 and for another three mAbs the neutralization specificity could not be determined.

Two types of mAbs appeared to be overrepresented: ten mAbs used VH 2-70 IGHV paired with VL {lambda}1-40, having characteristic mutations in complementarity determining region two (CDRL2). Cryogenic electron microscopy (Cryo-EM) revealed that two of these antibodies bound five Fabs per pentamer interacting with all five L1-surface loops. The other type of mAbs that appeared to be overrepresented were ten mAbs using VH 4-34, seven of which also used DH3-16*02 with conserved CDRH3 sequences. Cryo-EM for one of these mAbs, that required the FG-loop for neutralization, was shown to bind one Fab per pentamer at the apex, interacting with the DE- and FG-loops, with sequences of the Fab CDRH3 inserted between the DE- and FG-loops from two protomers. These two types of mAbs were found repeatedly in the four participants suggesting that these antibodies shared developmental pathways and bound to similar immunodominant epitopes on the virus.

HighlightsMost human mAbs recognized L1 surface loops but three of 70 recognized sequences on the C-terminal arm of L1

Some antibodies induced by HPV vaccination follow shared developmental pathways.

Human monoclonal antibodies using VH 2-70/VL {lambda}1-40 were found in all participants and bound with at a stoichiometry of five Fabs per capsomer.

Human monoclonal antibodies using the diversity gene segment D3-16*02 were found in all participants and one Fab was shown to bind with a stoichiometry of one Fab per capsomer.

In briefA panel of 70 HPV16 specific human monoclonal antibodies (mAbs), cloned from memory B cells or plasmablasts following HPV vaccination, was characterized by determining the surface loops of the major capsid protein (L1) required for neutralization and examined for shared molecular features. All five L1 loops were found to be used for neutralization by one or more antibodies, but the most frequent target of these antibodies was the FG loop followed by the HI and DE loops. Ten antibodies paired the heavy chain variable gene VH 2-70 with the light chain variable gene VL {lambda}1-40 and these antibodies had conserved mutations in the CDRL2 region of VL {lambda}1-40. Mutating the CDRL2 back to the predicted germline sequence significantly reduced neutralization. Cryo-EM analysis of two VH2-70/VL{lambda}1-40 mAbs showed five Fabs binding per L1 pentamer and a conserved epitope with Fabs interacting with all five variable loops across two adjacent protomers. Seven other mAbs had a heavy chain composed of the variable region VH4-34 with the diversity gene D3-16*02 resulting in the sequence motif WSGYR in the CDRH3. Mutation of that sequence to alanine ablated HPV16 neutralization activity. A cryo-EM structure of one of these antibodies showed one Fab binding the pentamer apex with the WSGYR motif inserting between three loops from two protomers. Antibodies with paired VH 2-70/VL {lambda}1-40 and the antibodies with CDRH3 containing the WSGYR sequence, were found in all four study participants suggesting that such antibodies may be commonly elicited following HPV vaccination.
]]></description>
<dc:creator>Carter, J. J.</dc:creator>
<dc:creator>Hurlburt, N. K.</dc:creator>
<dc:creator>Scherer, E. M.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Rodarte, J. V.</dc:creator>
<dc:creator>Smith, R. A.</dc:creator>
<dc:creator>Lewis, P.</dc:creator>
<dc:creator>Kinzelman, R. J.</dc:creator>
<dc:creator>Kieltyka, J.</dc:creator>
<dc:creator>Caban, M. E.</dc:creator>
<dc:creator>Wipf, G. C.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>Galloway, D. A.</dc:creator>
<dc:date>2025-04-07</dc:date>
<dc:identifier>doi:10.1101/2025.03.31.646278</dc:identifier>
<dc:title><![CDATA[HPV16 neutralizing monoclonal antibodies show evidence for common developmental pathways and public epitopes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.06.647354v1?rss=1">
<title>
<![CDATA[
Targeting the ITGA3B1 Complex with A Novel Antibody-Drug Conjugate for Metastatic Bladder Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.06.647354v1?rss=1</link>
<description><![CDATA[
Advances in antibody (Ab) phage display technologies have enabled the development of targeted therapies with enhanced precision and clinical efficacy. Here, we report a phenotypic screening strategy combining target-unbiased live-cell biopanning with in situ chemical crosslinking and mass spectrometry to identify internalizing antibodies and their cognate antigens--key features for effective antibody-drug conjugate (ADC) development. Using this approach, we identified the 2E7 antibody, which exhibits rapid internalization and high specificity for the integrin 3{beta}1 (ITGA3B1) heterodimer, a complex overexpressed in multiple solid tumors, including bladder cancer. We generated an ITGA3B1-targeted ADC by conjugating monomethyl auristatin E (MMAE) to the 2E7 antibody, enabling selective delivery of cytotoxic payloads to ITGA3B1-positive cells. In preclinical bladder cancer models, this ADC demonstrated potent and dose-dependent antitumor efficacy, with significant tumor regression and improved survival. Our findings establish a framework for target discovery using live-cell phenotypic screening and position ITGA3B1 as a promising therapeutic target for ADC-based treatment of advanced bladder cancer.

TeaserLive-cell biopanning uncovers ITGA3B1 as a target for ADC development in metastatic bladder cancer.
]]></description>
<dc:creator>Jung, H.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Kim, E. G.</dc:creator>
<dc:creator>Ahn, J.</dc:creator>
<dc:creator>Haffner, M.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:creator>Yi, E. C.</dc:creator>
<dc:creator>Kim, K. M.</dc:creator>
<dc:date>2025-04-07</dc:date>
<dc:identifier>doi:10.1101/2025.04.06.647354</dc:identifier>
<dc:title><![CDATA[Targeting the ITGA3B1 Complex with A Novel Antibody-Drug Conjugate for Metastatic Bladder Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.04.647286v1?rss=1">
<title>
<![CDATA[
Mucosal tissue NK cells tune their function between optimal anti-pathogen activity and tissue protection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.04.647286v1?rss=1</link>
<description><![CDATA[
Preserving barrier integrity while mounting effective immunity is essential at mucosal surfaces. In examining the immune cells that mediate both inflammation and tissue homeostasis, we uncovered a dual role for natural killer (NK) cells in barrier immunity. While NK cells are known to control viral infections, here we identify a previously unrecognized reparative function for NK cells in mucosal tissues. Using single-cell RNA sequencing and high-dimensional flow cytometry, we reveal a distinct population of human mucosal NK cells marked by tissue residency, immunoregulatory profiles, and limited cytotoxic potential at homeostasis, yet highly responsive to inflammatory cues. In a mouse model of acute HSV-2 infection, NK cells are required for limiting infection-associated tissue damage. Mucosal NK cells express the epithelial growth factor amphiregulin (Areg), and their depletion leads to increased tissue barrier damage despite preserved viral clearance, suggesting a novel role in tissue protection. Mechanistically, we demonstrate that the barrier-derived cytokines IL-18 and IL-33 induce Areg expression by both human and mouse NK cells, linking local inflammatory cues to reparative NK cell programming that is able to potentiate wound healing. Together, our findings reveal a context-dependent, dual function of mucosal NK cells in immune defense and mucosal tissue protection, expanding current models of NK cell biology.
]]></description>
<dc:creator>Vick, S. C.</dc:creator>
<dc:creator>Domenjo-Vila, E.</dc:creator>
<dc:creator>Frutoso, M.</dc:creator>
<dc:creator>Glabman, R. A.</dc:creator>
<dc:creator>Warrier, L. S.</dc:creator>
<dc:creator>Hughes, S. M.</dc:creator>
<dc:creator>Kirby, A. C.</dc:creator>
<dc:creator>Fialkow, M. F.</dc:creator>
<dc:creator>Hladik, F.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:creator>Lund, J. M.</dc:creator>
<dc:date>2025-04-09</dc:date>
<dc:identifier>doi:10.1101/2025.04.04.647286</dc:identifier>
<dc:title><![CDATA[Mucosal tissue NK cells tune their function between optimal anti-pathogen activity and tissue protection]]></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/10.1101/2025.04.04.647316v1?rss=1">
<title>
<![CDATA[
C-terminal fusion partner activity contributes to the oncogenic functions of YAP1::TFE3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.04.647316v1?rss=1</link>
<description><![CDATA[
YAP1 gene fusions are found in a multitude of human tumors, are potent oncogenic drivers, and are the likely initiating tumorigenic events in these tumors. We and others have previously shown that a YAP1 fusion proteins exert TEAD-dependent oncogenic YAP1 activity that is resistant to inhibitory Hippo pathway signaling. However, the contributions of the C-terminal fusion partners to the oncogenic functions of YAP1 fusion proteins are understudied. Here, we used the RCAS/tv-a system to express eight different YAP1 gene fusions in vivo and observed significant differences in the latencies of tumors induced by the various YAP1 fusions. We observed that tumors induced by YAP1::TFE3 displayed a significantly different histomorphology compared to tumors induced by other YAP1 fusions or activated non-fusion YAP1. To assess the extent to which the functional TFE3 domains (DNA binding: leucine zipper (LZ) and basic-helix-loop-helix (bHLH); activation domain (AD)) contribute to the oncogenic functions of YAP1::TFE3, we generated several mutant variants and performed functional in vitro and in vivo assays. In vitro, mutation or deletion of the TFE3 DNA binding domains (LZ, bHLH) resulted in reduced TFE3 activity but increased YAP1 activity of YAP1::TFE3. In vivo, deletion of the LZ and bHLH domains did not result in a decrease in tumor incidence but induced the formation of more YAP1-like tumors that lacked prominent features of YAP1::TFE3-driven tumors. By contrast, loss of the TFE3 AD almost completely abrogated tumor formation. Our results suggest that the TFE3 domains significantly contribute to the oncogenic activity of YAP1::TFE3.
]]></description>
<dc:creator>Cimino, P. J.</dc:creator>
<dc:creator>Keiser, D. J.</dc:creator>
<dc:creator>Parrish, A. G.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:date>2025-04-09</dc:date>
<dc:identifier>doi:10.1101/2025.04.04.647316</dc:identifier>
<dc:title><![CDATA[C-terminal fusion partner activity contributes to the oncogenic functions of YAP1::TFE3]]></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/10.1101/2025.04.14.648806v1?rss=1">
<title>
<![CDATA[
Designed NGF mimetics with reduced nociceptive signatures in neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.14.648806v1?rss=1</link>
<description><![CDATA[
The clinical use of Nerve Growth Factor (NGF) for neuronal regeneration has been hampered by pain sensitization side effects. NGF signals through the receptor tyrosine kinase TrkA and the co-receptor p75NTR; pain sensitization is thought to involve p75NTR. We sought to overcome this limitation by de novo design of a TrkA agonist that does not bind p75NTR. We designed homodimeric TrkA engaging constructs that dimerize TrkA subunits in a variety of geometries, and identified those eliciting the strongest signaling. The resulting designed agonists are able to stimulate transdifferentiated neurons and neuroblastoma cell lines, leading to neurite outgrowth and neuronal differentiation, with considerably reduced transcription of inflammation and pain related genes. These agonists are promising candidates for promoting neuronal regeneration without adverse side effects.

HighlightsO_LIDe novo designed TrkA agonists activate MAPK and PI3K-AKT signaling
C_LIO_LIRigid fusions allow for highly tunable signaling signatures
C_LIO_LITrkA agonists lead to neurite outgrowth in neuroblastoma cells comparable to retinoic acid
C_LIO_LIModulation of the TrkA pathway without co-stimulating p75NTR leads to a downregulation of inflammatory and nociceptive signature in neurons.
C_LI

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC="FIGDIR/small/648806v1_ufig1.gif" ALT="Figure 1">
View larger version (32K):
org.highwire.dtl.DTLVardef@c48066org.highwire.dtl.DTLVardef@c9c5a7org.highwire.dtl.DTLVardef@cf8ebdorg.highwire.dtl.DTLVardef@a4345a_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Schlichthaerle, T.</dc:creator>
<dc:creator>Yang, A.</dc:creator>
<dc:creator>Detraux, D.</dc:creator>
<dc:creator>Johnson, D. E.</dc:creator>
<dc:creator>Peach, C. J.</dc:creator>
<dc:creator>Edman, N. I.</dc:creator>
<dc:creator>Sniezek, C.</dc:creator>
<dc:creator>Williams, C. A.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Katiyar, N.</dc:creator>
<dc:creator>Chen, I.</dc:creator>
<dc:creator>Etemadi, A.</dc:creator>
<dc:creator>Favor, A.</dc:creator>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Kubo, C.</dc:creator>
<dc:creator>Coventry, B.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Gerben, S.</dc:creator>
<dc:creator>Ennist, N. M.</dc:creator>
<dc:creator>Milles, L.</dc:creator>
<dc:creator>Sankaran, B.</dc:creator>
<dc:creator>Kang, A.</dc:creator>
<dc:creator>Nguyen, H.</dc:creator>
<dc:creator>Bera, A. K.</dc:creator>
<dc:creator>Negahdari, B.</dc:creator>
<dc:creator>Hamazaki, N.</dc:creator>
<dc:creator>Schweppe, D. K.</dc:creator>
<dc:creator>Stewart, L.</dc:creator>
<dc:creator>Young, J. E.</dc:creator>
<dc:creator>Bunnett, N. W.</dc:creator>
<dc:creator>Ruohola-Baker, H.</dc:creator>
<dc:creator>Mathieu, J.</dc:creator>
<dc:creator>Pattwell, S.</dc:creator>
<dc:creator>Garcia, K. C.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2025-04-14</dc:date>
<dc:identifier>doi:10.1101/2025.04.14.648806</dc:identifier>
<dc:title><![CDATA[Designed NGF mimetics with reduced nociceptive signatures in neurons]]></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/10.1101/2025.04.10.648245v1?rss=1">
<title>
<![CDATA[
Modulating AP-1 enables CAR-T cells to establish an intratumoral PD-1+Tcf1+ stem-like reservoir and overcomes resistance to PD-1 axis blockade 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.10.648245v1?rss=1</link>
<description><![CDATA[
PD-1+Tcf1+ stem-like cells are critical mediators of endogenous T cell responses to PD-1/PD-L1 blockade and are maintained by MHC-dependent interactions with professional antigen-presenting cells (APCs). Unlike conventional T cells, CAR-T cells are activated by intact antigen expressed on tumors, not by peptide/MHC expressed on APCs, restricting their activation to the hostile tumor microenvironment (TME) that may impair preservation of this critical stem-like subset. Indeed, in an autochthonous model of ROR1+ lung cancer that we developed, CAR-T cells targeting the tumor-associated antigen ROR1 were uniformly Tcf1+ prior to infusion but rapidly downregulated Tcf1 in vivo and became terminally exhausted, similar to observations in patients, resulting in faster attrition and no enhancement in response to PD-L1 blockade. We hypothesized that overexpression of AP-1 family transcription factors, which can regulate T cell exhaustion, could enable CAR-Ts to maintain this critical PD-1+Tcf1+ subset within tumors independently of APCs and sensitize them to PD-1/PD-L1 blockade. Overexpression of the AP-1 TF c-Jun, but not BATF, improved preservation of PD-1+Tcf1+ CAR-T cells within tumors in a cell-intrinsic manner that correlated with increased persistence deeper within tumors. Notably, c-Jun overexpression alone was insufficient to prevent CAR-T exhaustion in the lung TME, in contrast to prior work in xenograft models, with progressive CAR-T dysfunction correlated with PD-1-dependent downregulation of c-Jun. However, c-Jun overexpression dramatically sensitized CAR-Ts to PD-L1 blockade, which restored c-Jun levels in CAR-Ts, drove log-fold expansion of CAR-Ts within tumors, and induced nearly complete eradication of ROR1+ tumor in highly aggressive models of lung cancer. Altogether, our data show that combination with PD-L1 blockade is necessary to unleash the full potential of c-Jun-overexpressing CAR-T cells in aggressive solid tumors like lung cancer and suggest that strategies to enhance formation of intratumoral PD-1+Tcf1+ reservoirs can overcome CAR-T resistance to PD-1 blockade.
]]></description>
<dc:creator>Snyder, A. J.</dc:creator>
<dc:creator>Shasha, C.</dc:creator>
<dc:creator>Ho, T.</dc:creator>
<dc:creator>Garrison, S.</dc:creator>
<dc:creator>Wilhelm, A. R.</dc:creator>
<dc:creator>Kluesner, M. G.</dc:creator>
<dc:creator>Ortiz, S.</dc:creator>
<dc:creator>Nutt, W. S.</dc:creator>
<dc:creator>Bingham, E.</dc:creator>
<dc:creator>Bhise, S. S.</dc:creator>
<dc:creator>Fan, E.</dc:creator>
<dc:creator>Zepeda, V.</dc:creator>
<dc:creator>Sarvothama, M.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Potluri, S.</dc:creator>
<dc:creator>Long, A.</dc:creator>
<dc:creator>Elz, A.</dc:creator>
<dc:creator>Furlan, S.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Srivastava, S.</dc:creator>
<dc:date>2025-04-15</dc:date>
<dc:identifier>doi:10.1101/2025.04.10.648245</dc:identifier>
<dc:title><![CDATA[Modulating AP-1 enables CAR-T cells to establish an intratumoral PD-1+Tcf1+ stem-like reservoir and overcomes resistance to PD-1 axis blockade]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.15.648856v1?rss=1">
<title>
<![CDATA[
Muc6-expressing gastric isthmus progenitors contribute to regeneration and metaplasia supported by myeloid-mesenchymal interactions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.15.648856v1?rss=1</link>
<description><![CDATA[
Gastric mucosal homeostasis is maintained by tissue-resident stem and progenitor cells residing in the isthmus region. Following mucosal injury, surviving cells contribute to regeneration, coinciding with characteristic pathological changes such as atrophic gastritis and metaplasia. To comprehensively understand the cellular dynamics involved in this process, we performed single-cell and spatial transcriptomics using newly generated transgenic mice. In human samples and mouse models, loss of gastric chief cells precedes, and even induces, loss of parietal cells during the progression of atrophy and metaplasia, validating the causal relationship underlying the decrease of these two lineages. Single-cell analysis confirmed robust stemness and metaplastic changes in the Muc6-expressing neck lineage following either chief or parietal cell ablation, and lineage-tracing experiments revealed that Muc6-expressing isthmus progenitors serve as a source of metaplasia and regeneration. Mechanistically, mucosal injury recruits IL-1-expressing myeloid cells, which stimulates NRG1 production in stromal fibroblasts, leading to mucosal proliferation and regeneration mediated by Myc activation in isthmus progenitors. These findings highlight the injury-responsible stem cell-like function of Muc6-expressing isthmal progenitors, which play a critical role in mucosal homeostasis and disease progression.

Visual abstract

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]]></description>
<dc:creator>Arai, J.</dc:creator>
<dc:creator>Iwata, Y.</dc:creator>
<dc:creator>Tsubosaka, A.</dc:creator>
<dc:creator>Kinoshita, H.</dc:creator>
<dc:creator>Shinohara, S.</dc:creator>
<dc:creator>Abe, S.</dc:creator>
<dc:creator>Shiokawa, T.</dc:creator>
<dc:creator>Oura, K.</dc:creator>
<dc:creator>Suzuki, N.</dc:creator>
<dc:creator>Hata, M.</dc:creator>
<dc:creator>Kurokawa, K.</dc:creator>
<dc:creator>Oya, Y.</dc:creator>
<dc:creator>Tsuboi, M.</dc:creator>
<dc:creator>Ihara, S.</dc:creator>
<dc:creator>Murakami, K.</dc:creator>
<dc:creator>Shiomi, C.</dc:creator>
<dc:creator>Uekura, C.</dc:creator>
<dc:creator>Fujiwara, H.</dc:creator>
<dc:creator>Tateno, H.</dc:creator>
<dc:creator>Mizuno, S.</dc:creator>
<dc:creator>Takahashi, S.</dc:creator>
<dc:creator>Kanai, A.</dc:creator>
<dc:creator>Suzuki, Y.</dc:creator>
<dc:creator>Ushiku, T.</dc:creator>
<dc:creator>Ijichi, H.</dc:creator>
<dc:creator>Hirata, Y.</dc:creator>
<dc:creator>Kasuga, M.</dc:creator>
<dc:creator>O'Brien, V. P.</dc:creator>
<dc:creator>Salama, N.</dc:creator>
<dc:creator>Kakiuchi, M.</dc:creator>
<dc:creator>Ishikawa, S.</dc:creator>
<dc:creator>WANG, T. C.</dc:creator>
<dc:creator>Hayakawa, Y.</dc:creator>
<dc:creator>Fujishiro, M.</dc:creator>
<dc:date>2025-04-16</dc:date>
<dc:identifier>doi:10.1101/2025.04.15.648856</dc:identifier>
<dc:title><![CDATA[Muc6-expressing gastric isthmus progenitors contribute to regeneration and metaplasia supported by myeloid-mesenchymal interactions]]></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/10.1101/2025.04.17.649396v1?rss=1">
<title>
<![CDATA[
Measurement and prediction of unmixing-dependent spreading in spectral flow cytometry panels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.17.649396v1?rss=1</link>
<description><![CDATA[
Advances in spectral cytometry instrumentation and fluorescent reagents have led to the possibility of ultra-high-parameter panels exceeding 50 colors. However, panel size is limited in practice by unmixing-dependent spreading (UDS), a mathematical phenomenon which leads to a progressive deterioration of unmixed signal-to-noise ratios in panels that contain fluorochrome combinations with significant spectral overlap. Choosing spectrally compatible sets of fluorochromes that avoid UDS is a complex and labor-intensive task involving substantial trial-and-error experimentation. Here, we provide a detailed explanation of UDS and practical strategies for handling UDS in large spectral panels. We describe the empirical hallmarks of UDS, demonstrate how to quantify its impact, and dissect its underlying mathematical cause in terms of spectral collinearity. We present novel computational metrics that can be used to select optimal combinations of fluorochromes in a platform-agnostic fashion based on publicly available reference data, providing a general tool for spectral panel design.
]]></description>
<dc:creator>Mage, P. L.</dc:creator>
<dc:creator>Konecny, A. J.</dc:creator>
<dc:creator>Mair, F.</dc:creator>
<dc:date>2025-04-23</dc:date>
<dc:identifier>doi:10.1101/2025.04.17.649396</dc:identifier>
<dc:title><![CDATA[Measurement and prediction of unmixing-dependent spreading in spectral flow cytometry panels]]></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/10.1101/2025.04.21.649828v1?rss=1">
<title>
<![CDATA[
Whole-Proteome Screening and Multi-Modal Profiling of Antigen-Specific CD4+ T Cells at Single-Cell Resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.21.649828v1?rss=1</link>
<description><![CDATA[
Systematic whole-proteome screening and comprehensive profiling of antigen-specific CD4+ T cells are crucial for advancing our understanding of CD4+ T cell immunity, yet such efforts remain technically challenging. Here, we present a high-throughput platform that employs large-scale class II single-chain trimer libraries to detect antigen-specific CD4+ T cells, while simultaneously profiling their antigen specificity, TCR/{beta} sequences, MHC restriction, whole transcriptomes, and patient/timepoint origins at single-cell resolution. We benchmarked SCTs against conventional pMHCs and validated the SCT library-based approach in direct ex vivo identification of antigen-specific CD4+ T cells in healthy donors. We then applied the platform to screen the entire SARS-CoV-2 receptor-binding domain in a longitudinal patient cohort, identifying 2,188 antigen-specific CD4+ T cells and revealing key features that define antigen immunogenicity. Extending to cancer, we performed whole-proteome screening of HPV-16 E6/E7 for TCR repertoire profiling in a precancerous cohort, uncovering functional heterogeneity of HPV-specific TCRs. By integrating high-throughput antigen screening with high-dimensional, multi-modal cellular characterization, our approach offers an unprecedented window into CD4+ T cell immunity across diverse disease contexts and empowers the development of new therapies.
]]></description>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Qi, J.</dc:creator>
<dc:creator>McKasson, M.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Gutierrez, V.</dc:creator>
<dc:creator>Brennan, C.</dc:creator>
<dc:creator>Hong, S.</dc:creator>
<dc:creator>Chour, W.</dc:creator>
<dc:creator>Ng, R. H.</dc:creator>
<dc:creator>Xie, J.</dc:creator>
<dc:creator>Yuan, D.</dc:creator>
<dc:creator>Webster, A.</dc:creator>
<dc:creator>Sidhu, S. K.</dc:creator>
<dc:creator>Anderson, A.</dc:creator>
<dc:creator>Chen, D.</dc:creator>
<dc:creator>Edmark, R.</dc:creator>
<dc:creator>Murray, K. M.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>McDonald, C.</dc:creator>
<dc:creator>Rowen, L.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Rasheed, Y.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Wagner, J. R.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Narwaly, K.</dc:creator>
<dc:creator>Fu, J.</dc:creator>
<dc:creator>Duven, A.</dc:creator>
<dc:creator>Forman, S. J.</dc:creator>
<dc:creator>Song, M.</dc:creator>
<dc:creator>Priceman, S.</dc:creator>
<dc:creator>Brown, C. E.</dc:creator>
<dc:creator>Ribas, A.</dc:creator>
<dc:creator>Wong, D.</dc:creator>
<dc:creator>Paulson, K. G.</dc:creator>
<dc:creator>Drescher, C. W.</dc:creator>
<dc:creator>Puig-Saus, C.</dc:creator>
<dc:creator>Goldman, J. D.</dc:creator>
<dc:creator>Trimble, C. L.</dc:creator>
<dc:creator>Heath, J. R.</dc:creator>
<dc:date>2025-04-23</dc:date>
<dc:identifier>doi:10.1101/2025.04.21.649828</dc:identifier>
<dc:title><![CDATA[Whole-Proteome Screening and Multi-Modal Profiling of Antigen-Specific CD4+ T Cells at Single-Cell Resolution]]></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/10.1101/2025.04.22.649918v1?rss=1">
<title>
<![CDATA[
Interprotomer Crosstalk in Mosaic Viral Glycoprotein Trimers Provides Insight into Polyvalent Immunogen Co-assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.22.649918v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 variants have demonstrated the ability to evade immune responses, leading to waves of infection throughout the pandemic. In response, bivalent mRNA vaccines, encoding the original Wuhan-Hu-1 and emerging variants, were developed to display both spike antigens. To date, it has not been determined whether co-transfection and co-translation of different SARS-CoV-2 variants results in co-assembly of mosaic heterotrimer antigens and how this may affect trimer stability, dynamics, and antigenicity. Understanding whether such mosaic heterotrimers can form and their implications for antigen structure can provide important information to guide future polyvalent vaccine design where multiple variants of an antigen are co-formulated. To investigate this, we purified mosaic spike assemblies of both genetically close (Omicron BA.2 and XBB) and distant (Omicron BA.2 and Wuhan-Hu-1 G614) strains. We found that the stability and integrity of mosaic spike trimers were maintained without misfolding or aggregation. Glycosylation profiles likewise were preserved relative to the homotrimer counterparts. Hydrogen/deuterium-exchange mass spectrometry and biolayer-interferometry were used to investigate the mosaic spike dynamics and any impact on epitope presentation and receptor binding. The Omicron-XBB heterotrimer, sharing a common fusion subunit sequence, retained protomer-specific dynamics similar to the corresponding homotrimers in antigenically important regions. The Omicron-G614 heterotrimer, co-assembling from protomers of divergent fusion subunit sequences, likewise showed overall similar dynamics and conformations in the receptor-binding subunit compared to the homotrimers. However, the incorporation of the Wuhan-Hu-1 G614 protomer led to a stabilizing effect on the relatively unstable Omicron fusion subunit in the heterotrimer. These findings reveal structural dynamic crosstalk in mosaic trimers, suggesting a potential for enhanced immunogen display and important considerations to be aware of in the use of polyvalent nucleic acid vaccines.

Author summaryHere we investigated the possibility of immunogen co-assembly from bivalent nucleic acid vaccines with a focus on probing potential impacts on antigen stability and epitope display. We purified mosaic heterotrimers composed of Omicron BA.2 and Wuhan-Hu-1 G614 protomers as well as those composed of Omicron BA.2 and XBB protomers. Both mosaic heterotrimers maintained their structural integrity, morphology, and N-glycosylation profiles. Using hydrogen/deuterium-exchange mass spectrometry, we demonstrated that both types of heterotrimers preserved strain-specific dynamics in critical antigenic regions, comparable to their homotrimer counterparts. Crosstalk between fusion subunits of Omicron BA.2 and Wuhan-Hu-1 G614 revealed a stabilizing effect of the G614 protomer on the inherently less stable fusion subunit in Omicron. Our findings provide insight into the structural dynamic profiles of co-expressed immunogen display, highlighting important considerations for polyvalent formulations that are being pursued to provide broad protection against highly variable pathogens.
]]></description>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Lovendahl, K.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:creator>Lee, K. K.</dc:creator>
<dc:date>2025-04-23</dc:date>
<dc:identifier>doi:10.1101/2025.04.22.649918</dc:identifier>
<dc:title><![CDATA[Interprotomer Crosstalk in Mosaic Viral Glycoprotein Trimers Provides Insight into Polyvalent Immunogen Co-assembly]]></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/10.1101/2025.05.02.651933v1?rss=1">
<title>
<![CDATA[
In vivo production of an anti-HIV antibody from primate hematopoietic cells by non-viral knock-in 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.02.651933v1?rss=1</link>
<description><![CDATA[
Non-viral gene editing offers a practical alternative to viral delivery for durable biologics production. Clinical trials have shown that adeno-associated virus encoding broadly neutralizing antibodies can protect against HIV, but result in limited, short-lived responses. The development of non-viral gene editing approaches in hematopoietic stem and progenitor cells holds promise for long-term antibody production. In this study, we evaluated CRISPR/Cas9 and CRISPR/Cas12a for gene knock-in at the immunoglobulin heavy chain locus in non-human primate hematopoietic stem and progenitor cells. Delivering the nuclease as a protein alongside a custom DNA template, we optimized editing with Cas12a and demonstrated higher knock-in efficiency and fewer non-specific edits than Cas9. Transplantation of edited non-human primate hematopoietic stem and progenitor cells into MISTRG mice led to engraftment, B cell differentiation, and transgene expression of a reporter transgene or anti-HIV antibody after HIV immunization with detectable anti-HIV antibody titers in peripheral blood circulation. These findings demonstrate the feasibility of using non-viral gene editing in HSPC as a potential strategy for sustained biologics production in the treatment of chronic diseases such as HIV. Future work will assess the efficacy of this model in a non-human primate model of HIV infection.
]]></description>
<dc:creator>Castelli, J. M.</dc:creator>
<dc:creator>Poljakov, K.</dc:creator>
<dc:creator>Youngseo, J.</dc:creator>
<dc:creator>Cassidy, M. E.</dc:creator>
<dc:creator>Gray, M. D.</dc:creator>
<dc:creator>Sanchez Gaytan, J. N.</dc:creator>
<dc:creator>Enstrom, M. R.</dc:creator>
<dc:creator>Linton, J. D.</dc:creator>
<dc:creator>Rongvaux, A.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:creator>Adair, J. E.</dc:creator>
<dc:date>2025-05-08</dc:date>
<dc:identifier>doi:10.1101/2025.05.02.651933</dc:identifier>
<dc:title><![CDATA[In vivo production of an anti-HIV antibody from primate hematopoietic cells by non-viral knock-in]]></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/10.1101/2025.05.04.652110v1?rss=1">
<title>
<![CDATA[
Bone marrow lymphocyte dynamics during chemotherapy in pediatric acute myeloid leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.04.652110v1?rss=1</link>
<description><![CDATA[
A better understanding of lymphocyte dynamics during current treatment regimens in pediatric AML is urgently needed to understand whether the application of bispecific T-cell-engagers (BiTEs) during periods of low tumor burden could be a viable treatment strategy. In this study, we found that induction 1, comprising mitoxantrone-etoposide-cytarabine in nearly all patients (as part of the NOPHO-DBH AML-2012 protocol), led to preserved or increased relative lymphocyte abundances alongside marked blast reduction in most cases. This was accompanied by a shift towards higher T-cell fractions, potentially creating a favorable window for BiTE therapy. The absence of a correlation between blast reduction and lymphocyte changes suggests that chemotherapy exerts differential effects on the lymphocyte compartment. Despite the heterogeneity of agents used in induction 2, more than half of patients showed a decline in lymphocyte levels. Nonetheless, the increase in T- and B-cells observed in most patients from the NOPHO-AML 2004 cohort after induction 2 suggests that lymphocyte recovery at this treatment stage is not uniformly impaired. Our transcriptomic and ex vivo functional data align with preclinical findings in adult AML and provide a basis for further investigations in in vivo models and early clinical trials. Such efforts should prioritize novel BiTE constructs targeting multiple tumor-associated (e.g., NCT05673057) or tumor-specific antigens.
]]></description>
<dc:creator>Koedijk, J. B.</dc:creator>
<dc:creator>Barneh, F.</dc:creator>
<dc:creator>Meesters-Ensing, J. I.</dc:creator>
<dc:creator>van Tuil, M.</dc:creator>
<dc:creator>Sonneveld, E.</dc:creator>
<dc:creator>Lambo, S.</dc:creator>
<dc:creator>Perzolli, A.</dc:creator>
<dc:creator>Schweighart, E. K.</dc:creator>
<dc:creator>Ferrao Blanco, M. N.</dc:creator>
<dc:creator>van der Meulen, M.</dc:creator>
<dc:creator>Deli, A.</dc:creator>
<dc:creator>Haasjes, E.</dc:creator>
<dc:creator>Bang Christensen, K.</dc:creator>
<dc:creator>de Groot-Kruseman, H. A.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Hasle, H.</dc:creator>
<dc:creator>Belderbos, M.</dc:creator>
<dc:creator>Luesink, M.</dc:creator>
<dc:creator>Goemans, B. F.</dc:creator>
<dc:creator>Nierkens, S.</dc:creator>
<dc:creator>Hehir-Kwa, J.</dc:creator>
<dc:creator>Zwaan, C. M.</dc:creator>
<dc:creator>Heidenreich, O.</dc:creator>
<dc:date>2025-05-09</dc:date>
<dc:identifier>doi:10.1101/2025.05.04.652110</dc:identifier>
<dc:title><![CDATA[Bone marrow lymphocyte dynamics during chemotherapy in pediatric acute myeloid leukemia]]></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/10.1101/2025.05.13.653775v1?rss=1">
<title>
<![CDATA[
Intracellular and extracellular dynamics of herpes simplex virus 1 DNA and infectious particles in epithelial and neuronal cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.13.653775v1?rss=1</link>
<description><![CDATA[
Herpes simplex virus 1 (HSV-1) infection of epithelial cells is lytic, while infection of neurons typically results in long-term latency. However, the rates at which HSV-1 replicates and spreads in epithelial cells versus neurons under low and high multiplicity of infection (MOI) conditions remain undefined. Identifying these rates requires the application of mathematical models to carefully designed viral kinetic experiments. It is also critical to differentiate the dynamics of infectious viral particles versus viral DNA, as both quantities are routinely measured in in vitro experiments and human studies using plaque assays and polymerase chain reactions, respectively. Here, we developed mechanistic mathematical models to describe HSV-1 dynamics after infection of epithelial Vero cells and neuronal N2A cells, at high (3) and low (0.01) MOI. Our model recapitulates the dynamics of cell-free and cell-associated viral DNA and plaque-forming units (PFU). In epithelial cells, the model describes a pre-productive eclipse phase with a mean duration of 10.9 and 12.8 hours prior to HSV DNA replication and PFU production, respectively. Cells exited the eclipse phase as early and late as 2.5 and 32 hours, respectively. Infected cells produced a single PFU for every 224 HSV DNA genomes. PFU egressed at a constant rate, whereas the HSV DNA egress rate increased over time, before saturating at a 15 times higher rate. Under low relative to high MOI conditions, Vero cells spent 7 hours longer in the eclipse phase, had a 12-hour delay prior to egress, and had a longer mean duration of productive infection (14 versus 3.5-hour half-life). Secondary epithelial cell infection in low MOI experiments was overwhelmingly due to cell-to-cell viral spread and originated from a small number of early-producer cells. Neuronal cells produced viruses at a 5-fold lower rate and had a longer (mean: 42 hours) and more variable eclipse phase, with some neurons remaining in eclipse for more than a week. Our results highlighted large differences in HSV egress rates, as well as infected cell eclipse phase duration and death rates, in epithelial cells versus neurons during low and high MOI infection. The observed viral dynamics in neurons reflect a balance between active replication and latency.
]]></description>
<dc:creator>Esmaeili, S.</dc:creator>
<dc:creator>Swan, D. A.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Schiffer, J. T.</dc:creator>
<dc:creator>Walter, M.</dc:creator>
<dc:date>2025-05-13</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.653775</dc:identifier>
<dc:title><![CDATA[Intracellular and extracellular dynamics of herpes simplex virus 1 DNA and infectious particles in epithelial and neuronal cells]]></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/10.1101/2025.05.13.653810v1?rss=1">
<title>
<![CDATA[
Spatial control of secretory vesicle targeting by the Ync13-Rga7-Rng10 complex during cytokinesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.13.653810v1?rss=1</link>
<description><![CDATA[
Cytokinesis requires precise coordination of contractile-ring constriction, vesicle trafficking and fusion to the plasma membrane, and extracellular matrix assembly/remodeling at the cleavage furrow to ensure faithful cell division and maintain cell integrity. These processes and proteins involved are broadly conserved across eukaryotes, yet molecular mechanisms controlling the spatiotemporal pathways of membrane trafficking remain poorly understood. Here, using fission yeast genetics, microscopy, and in vitro binding assays, we identify a conserved module including the Munc13 protein Ync13, F-BAR protein Rga7, and coiled-coil protein Rng10 to be critical for precise and selective vesicle targeting during cytokinesis. The module specifically recruit the TRAPP-II but not exocyst complex to tether vesicles containing the glucan synthases Bgs4 and Ags1 along the cleavage furrow. Ync13 subsequently interacts with the SM protein Sec1 for vesicle fusion. Mutations in this pathway disrupt septum integrity and lead to cell lysis. Our work provides key insights into how membrane trafficking is tightly controlled to maintain cell integrity during cytokinesis.
]]></description>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Singh, D.</dc:creator>
<dc:creator>Zhu, Y.-H.</dc:creator>
<dc:creator>Zhang, K. J.</dc:creator>
<dc:creator>Melero, A.</dc:creator>
<dc:creator>Martin, S. G.</dc:creator>
<dc:creator>Wu, J.-Q.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.653810</dc:identifier>
<dc:title><![CDATA[Spatial control of secretory vesicle targeting by the Ync13-Rga7-Rng10 complex during cytokinesis]]></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/10.1101/2025.05.14.654106v1?rss=1">
<title>
<![CDATA[
Efficacy of DNA Methyltransferase Inhibitor Immune Priming Therapy in Combination with PD-1 Inhibitors to Treat High-Risk Pediatric Brain Tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.14.654106v1?rss=1</link>
<description><![CDATA[
BackgroundDespite intensive therapies, outcomes for high-risk pediatric brain tumors (PBTs) remain dismal, prompting the search for novel treatments. DNA methyltransferase inhibitors (DNMTi) have been shown to prime tumors to improve response to checkpoint inhibition. The aim of this study was to investigate the potential of decitabine (DAC), in combination with a PD-1 inhibitor, to improve survival in pediatric high-risk brain tumor models.

MethodsAnalysis of human PBT datasets was performed to determine gene expression levels of immune cell associated markers. Tumor response to DAC, with or without a PD-1 inhibitor, was tested in murine models representing H3-wildtype diffuse intrinsic pontine glioma (DIPG), H3K27-mutant diffuse midline glioma (DMG), atypical teratoid rhabdoid tumor (ATRT), and medulloblastoma (MB). CyTOF analysis of allograft tumors was performed to characterize changes within the tumor microenvironment.

ResultsAnalysis of PBT subtypes revealed heterogeneous expression of immune cell markers, checkpoint receptors, and MHC molecules. DAC treatment decreased DNA methylation and increased neoantigen expression in human and mouse tumor cells. DAC alone or in combination with a PD-1 inhibitor resulted in prolonged survival in syngeneic mouse models of DIPG and ATRT but not DMG and MB models. CyTOF analysis of mouse tumors revealed changes in local immune cell infiltration upon combination treatment.

ConclusionsDAC in combination with a PD-1 inhibitor can alter the immune microenvironment in mouse tumor models. Changes were observed in H3-wildtype DIPG and ATRT models, suggesting that certain tumor subtypes may respond to checkpoint blockade after immune augmentation with DNMTi.

Key PointsO_LIPBTs show heterogenous expression of immune cell infiltrates
C_LIO_LIDAC or DAC plus a PD-1 inhibitor shows extension of survival in H3-wildtype DIPG and ATRT mouse models
C_LIO_LIMyeloid-derived suppressor cell abundance could be a major contributing factor to treatment response
C_LI

Importance of the StudyChildren with high-risk PBTs face dismal outcomes. Immune checkpoint inhibitor (ICI) successes have been demonstrated in a variety of adult malignancies; however, such beneficial outcomes have not been realized in PBTs. Here we investigate single and combination treatment of DNMTi and PD-1 checkpoint inhibition in syngeneic mouse models of high-risk PBTs. Our results suggest that some H3-wildtype DIPG and ATRT tumor types may be responsive to checkpoint therapy post immunomodulation and warrant further investigation.
]]></description>
<dc:creator>Mishra, D. K.</dc:creator>
<dc:creator>Morris, S. M.</dc:creator>
<dc:creator>Popovski, D.</dc:creator>
<dc:creator>Girard, E. J.</dc:creator>
<dc:creator>Bondoc, A.</dc:creator>
<dc:creator>Kumar, S. S.</dc:creator>
<dc:creator>Andrade, A. F.</dc:creator>
<dc:creator>Zhu, X.</dc:creator>
<dc:creator>Yao, F.</dc:creator>
<dc:creator>Brusniak, M.-Y.</dc:creator>
<dc:creator>Umaru, B.</dc:creator>
<dc:creator>Crotty, E. E.</dc:creator>
<dc:creator>Brasel, K.</dc:creator>
<dc:creator>Pakiam, F.</dc:creator>
<dc:creator>Russo, C.</dc:creator>
<dc:creator>Zeinieh, M.</dc:creator>
<dc:creator>Biery, M. C.</dc:creator>
<dc:creator>Coxon, M.</dc:creator>
<dc:creator>Conti, H.</dc:creator>
<dc:creator>Clarke, M.</dc:creator>
<dc:creator>Lu, M.</dc:creator>
<dc:creator>Rutka, J.</dc:creator>
<dc:creator>Llivichuzhca-Loja, D.</dc:creator>
<dc:creator>Konnikova, L.</dc:creator>
<dc:creator>Fouladi, M.</dc:creator>
<dc:creator>Jabado, N.</dc:creator>
<dc:creator>Huang, A.</dc:creator>
<dc:creator>Olson, J. M.</dc:creator>
<dc:creator>Drissi, R.</dc:creator>
<dc:date>2025-05-18</dc:date>
<dc:identifier>doi:10.1101/2025.05.14.654106</dc:identifier>
<dc:title><![CDATA[Efficacy of DNA Methyltransferase Inhibitor Immune Priming Therapy in Combination with PD-1 Inhibitors to Treat High-Risk Pediatric Brain Tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.20.654130v1?rss=1">
<title>
<![CDATA[
Determining preclinical safety of Aclarubicin in pediatric malignancies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.20.654130v1?rss=1</link>
<description><![CDATA[
BackgroundAnthracyclines are among the most effective chemotherapeutic agents used to treat pediatric malignancies. However, their clinical use is limited by dose-dependent toxicities, particularly cardiotoxicity and secondary malignancies. Aclarubicin (Acla) is an anthracycline derivative that induces chromatin damage while sparing DNA, offering potential therapeutic benefit with reduced longterm toxicity.

MethodsWe evaluated the anti-tumor efficacy and safety profile of Acla in multiple in vitro pediatric cancer models and in vivo mouse models designed to mimic anthracycline re-treatment following prior doxorubicin (Doxo) exposure. Tumor growth, genotoxic stress, survival, and organ toxicity were assessed.

ResultsAcla demonstrated robust anti-tumor activity comparable to Doxo across diverse pediatric in vitro models. Unlike Doxo, Acla treatment did not induce significant genotoxic stress. In vivo, mice receiving Acla after Doxo exposure showed no evidence of cumulative cardiotoxicity or end-organ damage. In contrast, a second course of Doxo led to significant toxic mortality, but was surprisingly not attributable to classic cardiac injury.

ConclusionOur study highlights Acla as a promising anthracycline derivative for pediatric cancers, with potent anti-tumor efficacy and a superior safety profile, even following prior anthracycline exposure. These results support continued investigation of chromatin-damaging anthracyclines that can kill pediatric cancer cells without inducing genotoxic stress. In addition, our studies underscore the need to refine preclinical models to better understand both acute and chronic anthracycline toxicities in pediatric and adolescent populations.
]]></description>
<dc:creator>Tu, D. S.</dc:creator>
<dc:creator>Olson, A. K.</dc:creator>
<dc:creator>Waggie, K. S.</dc:creator>
<dc:creator>Garcia, N. M.</dc:creator>
<dc:creator>Hoglund, V. J.</dc:creator>
<dc:creator>Walter, S.</dc:creator>
<dc:creator>Rosinski, J. R.</dc:creator>
<dc:creator>Sadeeshkumar, H.</dc:creator>
<dc:creator>Patel, R.</dc:creator>
<dc:creator>Sayar, E.</dc:creator>
<dc:creator>Haffner, M.</dc:creator>
<dc:creator>Maves, L.</dc:creator>
<dc:creator>Neefjes, J.</dc:creator>
<dc:creator>Sarthy, J. F.</dc:creator>
<dc:creator>Lawlor, E. R.</dc:creator>
<dc:creator>Ganapathi, S. S.</dc:creator>
<dc:date>2025-05-24</dc:date>
<dc:identifier>doi:10.1101/2025.05.20.654130</dc:identifier>
<dc:title><![CDATA[Determining preclinical safety of Aclarubicin in pediatric malignancies]]></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/10.1101/2025.05.23.655786v1?rss=1">
<title>
<![CDATA[
Differential HIV-1 Proviral Defects in Children vs. Adults on Antiretroviral Therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.23.655786v1?rss=1</link>
<description><![CDATA[
HIV-1 proviral landscapes were investigated using near full-length HIV single-genome sequencing on blood samples from 5 children with vertically acquired infection and on ART for [~]7-9 years. Proviral structures were compared to published datasets in children prior to ART, children on short-term ART, and adults on ART. We found a strong selection for large internal proviral deletions in children, especially deletions of the env gene. Only 2.5% of the proviruses were sequence-intact, lower than in the comparative datasets from adults. Of the proviruses that retained the env gene, >80% contained two or more defects, most commonly stop codons and/or gag start mutations. Significantly fewer defects in the major splice donor site (MSD) and packaging signal were found in the children on short or long-term ART compared to the adults, and tat was more frequently defective in children. These results suggest that different selection pressures shape the proviral landscape in children compared to adults and reveal potentially different genetic regions to target for measuring the intact HIV reservoir and for achieving HIV remission in children.
]]></description>
<dc:creator>Hasson, J. M.</dc:creator>
<dc:creator>Katusiime, M. G.</dc:creator>
<dc:creator>Capoferri, A. A.</dc:creator>
<dc:creator>Bale, M. J.</dc:creator>
<dc:creator>Luke, B. T.</dc:creator>
<dc:creator>Shao, W.</dc:creator>
<dc:creator>Cotton, M. F.</dc:creator>
<dc:creator>Van Zyl, G.</dc:creator>
<dc:creator>Patro, S. C.</dc:creator>
<dc:creator>Kearney, M. F.</dc:creator>
<dc:date>2025-05-27</dc:date>
<dc:identifier>doi:10.1101/2025.05.23.655786</dc:identifier>
<dc:title><![CDATA[Differential HIV-1 Proviral Defects in Children vs. Adults on Antiretroviral Therapy]]></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/10.1101/2025.05.23.655879v1?rss=1">
<title>
<![CDATA[
Graphical and Interactive Spatial Proteomics Image Analysis Workflow 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.23.655879v1?rss=1</link>
<description><![CDATA[
Spatial proteomics provides a spatially resolved view of protein expression and localization within cells and tissues by mapping the location and abundance of proteins. There is a need for fully-integrated end-to-end imaging workflows for spatial proteomic analysis that are flexible, high-throughput, and support graphical and interactive visualizations. We present a modular and interactive spatial proteomic image analysis workflow with individual steps containerized that empowers biomedical researchers to reproducibly execute and customize complex analyses.

Our workflow consists of cell segmentation, unsupervised clustering, validation of clusters on the image, and cell type clustering results visualization. Users can utilize a form-based graphical interface to execute and customize multi-step workflows with a single click or interactively adjust image processing steps within the workflow, apply workflows to various datasets, and modify input parameters as needed. We illustrated the functionality of our workflow using a cancer imaging dataset consisting of a tissue microarray (TMA) stained by high-plex immunohistochemistry. This TMA contained a variety of cancer and tissue cell types to assess the broad applicability of this workflow to different biopsy and tissue types.
]]></description>
<dc:creator>Singh, P.</dc:creator>
<dc:creator>Wright, J. H.</dc:creator>
<dc:creator>Smythe, K. S.</dc:creator>
<dc:creator>Fukuda, B. N.</dc:creator>
<dc:creator>Hung, L.-H.</dc:creator>
<dc:creator>Yeung, C. C.</dc:creator>
<dc:creator>Yeung, K. Y.</dc:creator>
<dc:date>2025-05-27</dc:date>
<dc:identifier>doi:10.1101/2025.05.23.655879</dc:identifier>
<dc:title><![CDATA[Graphical and Interactive Spatial Proteomics Image Analysis Workflow]]></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/10.1101/2025.05.29.656244v1?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/10.1101/2025.05.27.656504v1?rss=1">
<title>
<![CDATA[
Engineering Sustained-Release Broadly Neutralizing Antibody Formulations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.27.656504v1?rss=1</link>
<description><![CDATA[
Sustained serum levels of broadly neutralizing antibodies (bnAbs) are crucial for effective passive immunization against infectious diseases as protection persists only while these bnAbs remain at adequate concentrations within the body. Current obstacles, such as poor pharmacokinetics (PK) and burdensome administration, must be overcome to make bnAbs a viable option for pre- and post-exposure prophylaxis. In this work, we explore how a polymer-nanoparticle (PNP) hydrogel depot technology can be engineered to prolong protein delivery. In-vivo studies in mice and rats demonstrate prolonged protein release, and modeling efforts predict the impact of both the elimination half-life of the active pharmaceutical ingredient and hydrogel depot volume on overall pharmacokinetics. Moreover, flow cytometry characterization reveals that immune cell infiltration into the hydrogel depot can result in faster-than-expected release of antibody cargo on account of active transport via cellular uptake. We then demonstrate that co-formulation of antibodies with an anti-inflammatory agent reduces cellular infiltration and resulting active transport, further extending delivery and pharmacokinetics. Finally, multicompartmental modeling predicts the human PK profiles of clinically relevant HIV bnAbs delivered via subcutaneous hydrogel injection. These findings aid in the development of next generation hydrogel materials that stabilize and slowly release bnAbs for long-term pre-exposure immunoprophylaxis.
]]></description>
<dc:creator>Jons, C. K.</dc:creator>
<dc:creator>Kasse, C. M.</dc:creator>
<dc:creator>Mayer, B. T.</dc:creator>
<dc:creator>Hyrien, O.</dc:creator>
<dc:creator>Sen, S.</dc:creator>
<dc:creator>Meany, E. L.</dc:creator>
<dc:creator>d'Aquino, A. I.</dc:creator>
<dc:creator>Ganesh, P.</dc:creator>
<dc:creator>Eckman, N.</dc:creator>
<dc:creator>Dong, C.</dc:creator>
<dc:creator>Yan, J.</dc:creator>
<dc:creator>Nguyen, L. T.</dc:creator>
<dc:creator>Doulames, V. M.</dc:creator>
<dc:creator>Song, Y. E.</dc:creator>
<dc:creator>Saouaf, O. M.</dc:creator>
<dc:creator>Williams, C. M.</dc:creator>
<dc:creator>Williams, S. C.</dc:creator>
<dc:creator>Paredes, J.</dc:creator>
<dc:creator>Raghavan, R.</dc:creator>
<dc:creator>Palomares, M.</dc:creator>
<dc:creator>Alpert, M.</dc:creator>
<dc:creator>Yates, N. L.</dc:creator>
<dc:creator>Tomaras, G.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Farzan, M.</dc:creator>
<dc:creator>Appel, E. A.</dc:creator>
<dc:date>2025-06-01</dc:date>
<dc:identifier>doi:10.1101/2025.05.27.656504</dc:identifier>
<dc:title><![CDATA[Engineering Sustained-Release Broadly Neutralizing Antibody Formulations]]></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/10.1101/2025.05.28.656673v1?rss=1">
<title>
<![CDATA[
OGDHL regulates nucleotide metabolism, tumor growth, and neuroendocrine marker expression in prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.28.656673v1?rss=1</link>
<description><![CDATA[
As cancer cells evade therapeutic pressure and adopt alternate lineage identities not commonly observed in the tissue of origin, they likely adopt alternate metabolic programs to support their evolving demands. Targeting these alternative metabolic programs in distinct molecular subtypes of aggressive prostate cancer may lead to new therapeutic approaches to combat treatment-resistance. We identify the poorly studied metabolic enzyme Oxoglutarate Dehydrogenase-Like (OGDHL), named for its structural similarity to the tricarboxylic acid (TCA) cycle enzyme Oxoglutarate Dehydrogenase (OGDH), as an unexpected regulator of tumor growth, treatment-induced lineage plasticity, and DNA Damage in prostate cancer. While OGDHL has been described as a tumor-suppressor in various cancers, we find that its loss impairs prostate cancer cell proliferation and tumor formation. Loss of OGDHL profoundly alters Androgen Receptor inhibition-induced plasticity, including suppressing the neuroendocrine markers DLL3 and HES6, induces accumulation of the DNA damage response marker {gamma}H2AX, and reduces nucleotide synthesis. Our data suggest that OGDHL has minimal impact on TCA cycle activity, and that mitochondrial localization is not required for its regulation of prostate cancer plasticity and nucleotide metabolism. Finally, we demonstrate that OGDHL expression is tightly correlated with neuroendocrine differentiation in clinical prostate cancer. These findings underscore the importance of investigating poorly characterized metabolic genes as potential regulators of distinct molecular subtypes of aggressive cancer.
]]></description>
<dc:creator>Bernard, M.</dc:creator>
<dc:creator>Ruiz, A.</dc:creator>
<dc:creator>Diaz, J.</dc:creator>
<dc:creator>Nunley, N.</dc:creator>
<dc:creator>Dove, R.</dc:creator>
<dc:creator>Heering, K.</dc:creator>
<dc:creator>Bopardikar, S.</dc:creator>
<dc:creator>Gallardo, A.</dc:creator>
<dc:creator>Hashimoto, T.</dc:creator>
<dc:creator>Agrawal, R.</dc:creator>
<dc:creator>Smith, C.</dc:creator>
<dc:creator>Wilde, B.</dc:creator>
<dc:creator>Matulionis, N.</dc:creator>
<dc:creator>Richards, H.</dc:creator>
<dc:creator>Sharifi, M.</dc:creator>
<dc:creator>Lang, J.</dc:creator>
<dc:creator>Zhao, S.</dc:creator>
<dc:creator>Haffner, M.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:creator>Christofk, H.</dc:creator>
<dc:creator>Goldstein, A. S.</dc:creator>
<dc:date>2025-06-01</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656673</dc:identifier>
<dc:title><![CDATA[OGDHL regulates nucleotide metabolism, tumor growth, and neuroendocrine marker expression in prostate cancer]]></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/10.1101/2025.05.28.656535v1?rss=1">
<title>
<![CDATA[
Chorionic Gonadotropin Beta 7 is a marker of immune evasion in cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.28.656535v1?rss=1</link>
<description><![CDATA[
Human chorionic gonadotropin beta (beta-hCG) is an oncofetal antigen expressed by trophoblast cells of the placenta, with minimal expression in adult somatic tissues. Numerous studies have demonstrated that beta-hCG-encoding genes are expressed in various cancers, but expression of these genes (CGB3, CGB5, CGB7, and CGB8) across diverse cancers has not been systematically evaluated. Here, we report that CGB genes are more widely expressed across diverse cancer types than previously appreciated and that secreted beta-hCG is readily detected. In particular, CGB genes are expressed in the majority of urothelial bladder cancers, where CGB7 is most frequently expressed and significantly associated with an immunosuppressed tumor microenvironment, including decreased CD8+ T cell infiltration. Multiple CGB genes are associated with failure to respond to immune checkpoint inhibitor (ICI) therapy, and CGB7 is particularly strongly predictive of poor prognosis. Overall, our findings indicate that beta-hCG is a clinically accessible, predictive biomarker of immunotherapeutic response.
]]></description>
<dc:creator>McKellar, S. A.</dc:creator>
<dc:creator>Pineda, J. M. B.</dc:creator>
<dc:creator>Lattupally, R.</dc:creator>
<dc:creator>Codd, A. S.</dc:creator>
<dc:creator>Newell, E.</dc:creator>
<dc:creator>Lu, S. X.</dc:creator>
<dc:creator>Bradley, R. K.</dc:creator>
<dc:date>2025-06-02</dc:date>
<dc:identifier>doi:10.1101/2025.05.28.656535</dc:identifier>
<dc:title><![CDATA[Chorionic Gonadotropin Beta 7 is a marker of immune evasion in cancer]]></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/10.1101/2025.06.01.657285v1?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/10.1101/2025.05.31.657155v1?rss=1">
<title>
<![CDATA[
Diverse modes of T cell receptor sequence convergence define unique functional and cellular phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.31.657155v1?rss=1</link>
<description><![CDATA[
Single-cell techniques allow concurrent study of gene activity and T cell receptor (TCR) sequences, identifying connections between TCR structure and cell traits. Expanding on our CoNGA software, we present a "metaCoNGA" analysis of 6 million T cells from 91 diverse studies, mapping TCR sequence similarity across tissues and diseases. This approach exposes shared TCR features within specific T cell subsets, including those associated with infection, cancer, and autoimmunity. We introduce a method to identify T cell groups with similar gene expression and biased TCR amino acid composition, providing a systematic framework for classifying diverse unconventional T cells, including KIR+ CD8+ T cells, CD4+ regulatory T cells, and subsets of NKT and MAIT cells. A new TCR clustering approach identifies thousands of convergent TCR sequence clusters hypothesized to target shared antigens. These clusters show coherent gene expression, highlighting the role of antigen exposure in shaping T cell behavior. Finally, we provide a tool for users to merge new data with this resource and rapidly identify T cell features in their data sets. This resource empowers investigations into the complex relationship between TCR sequence and T cell function in human health.
]]></description>
<dc:creator>Schattgen, S.</dc:creator>
<dc:creator>Vegesana, K.</dc:creator>
<dc:creator>Hazelton, W. D.</dc:creator>
<dc:creator>Minervina, A.</dc:creator>
<dc:creator>Valkiers, S.</dc:creator>
<dc:creator>Slowikowski, K.</dc:creator>
<dc:creator>Smith, N. P.</dc:creator>
<dc:creator>MGH COVID-19 Team,</dc:creator>
<dc:creator>Villani, A.-C.</dc:creator>
<dc:creator>Thomas, P. G.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:date>2025-06-03</dc:date>
<dc:identifier>doi:10.1101/2025.05.31.657155</dc:identifier>
<dc:title><![CDATA[Diverse modes of T cell receptor sequence convergence define unique functional and cellular phenotypes]]></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/10.1101/2025.06.02.657429v1?rss=1">
<title>
<![CDATA[
Lack of synergy between AR targeted therapies and PARP inhibitors in homologous recombination-proficient prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.02.657429v1?rss=1</link>
<description><![CDATA[
Recent clinical trials have explored the combination of androgen receptor (AR) pathway inhibitors and poly (ADP-ribose) polymerase (PARP) inhibitors as a potential treatment for castration-resistant prostate cancer. This combination treatment is based on the premise that AR directly regulates expression of DNA repair genes, leading to synergy between PARP and AR inhibition. Despite some promising preclinical evidence, this combination therapy has shown limited efficacy in patients with homologous recombination (HR)-proficient tumors. To investigate this discrepancy between preclinical and clinical results, we profiled the effects of PARP inhibition in prostate cancer models in the presence or absence of AR inhibition. Surprisingly, AR inhibition impaired response to PARP inhibitors in castration-sensitive cells and had no effect on response in castration-resistant cells. AR inhibition also did not regulate DNA repair in either the castration-resistant or castration-sensitive setting. Instead, we find that cell cycle progression is required for response to PARP inhibition in homologous-recombination proficient prostate cancer.

STATEMENT OF SIGNIFICANCEAndrogen deprivation does not inhibit DNA repair and does not synergize with PARP inhibition in prostate cancer with intact homologous recombination repair.
]]></description>
<dc:creator>Traphagen, N. A.</dc:creator>
<dc:creator>Wheeler, E.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Lu, F.</dc:creator>
<dc:creator>Ahmed, B.</dc:creator>
<dc:creator>Tewari, A. K.</dc:creator>
<dc:creator>Balk, S. P.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Corey, E.</dc:creator>
<dc:creator>Long, H.</dc:creator>
<dc:creator>D'Andrea, A. D.</dc:creator>
<dc:creator>Qiu, X.</dc:creator>
<dc:creator>Brown, M.</dc:creator>
<dc:date>2025-06-03</dc:date>
<dc:identifier>doi:10.1101/2025.06.02.657429</dc:identifier>
<dc:title><![CDATA[Lack of synergy between AR targeted therapies and PARP inhibitors in homologous recombination-proficient prostate cancer]]></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/10.1101/2025.06.02.656870v1?rss=1">
<title>
<![CDATA[
Replaying germinal center evolution on a quantified affinity landscape 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.02.656870v1?rss=1</link>
<description><![CDATA[
Darwinian evolution of immunoglobulin genes within germinal centers (GC) underlies the progressive increase in antibody affinity following antigen exposure. Whereas the mechanics of how competition between GC B cells drives increased affinity are well established, the dynamical evolutionary features of this process remain poorly characterized. We devised an experimental evolution model in which we "replay" over one hundred instances of a clonally homogenous GC reaction and follow the selective process by assigning affinities to all cells using deep mutational scanning. Our data reveal how GCs achieve predictable evolutionary outcomes through the cumulative effects of many rounds of imperfect selection, acting on a landscape shaped heavily by somatic hypermutation (SHM) targeting biases. Using time-calibrated models, we show that apparent features of GC evolution such as permissiveness to low-affinity lineages and early plateauing of affinity are best explained by survivorship biases that distort our view of how affinity progresses over time.
]]></description>
<dc:creator>DeWitt, W. S. S.</dc:creator>
<dc:creator>Vora, A. A.</dc:creator>
<dc:creator>Araki, T.</dc:creator>
<dc:creator>Galloway, J. G.</dc:creator>
<dc:creator>Alkutkar, T.</dc:creator>
<dc:creator>Bortolatto, J.</dc:creator>
<dc:creator>Castro, T. B.</dc:creator>
<dc:creator>Dumm, W.</dc:creator>
<dc:creator>Jennings-Schaffer, C.</dc:creator>
<dc:creator>Jia, T.</dc:creator>
<dc:creator>Mesin, L.</dc:creator>
<dc:creator>Ozorowski, G.</dc:creator>
<dc:creator>Pae, J.</dc:creator>
<dc:creator>Ralph, D. K.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Nourmohammad, A.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Ward, A.</dc:creator>
<dc:creator>Starr, T. N.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:date>2025-06-05</dc:date>
<dc:identifier>doi:10.1101/2025.06.02.656870</dc:identifier>
<dc:title><![CDATA[Replaying germinal center evolution on a quantified affinity landscape]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.03.657769v1?rss=1">
<title>
<![CDATA[
Comparing phenotypic manifolds with Kompot: Detecting differential abundance and gene expression at single-cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.03.657769v1?rss=1</link>
<description><![CDATA[
Single-cell studies are frequently designed to compare across conditions such as health and disease. However, existing computational approaches typically rely on grouping cells into discrete populations before making comparisons, which can limit resolution for detecting state-dependent changes. Here, we introduce Kompot, a statistical framework for comparative analysis of multi-condition single-cell data. Kompot quantifies both differential abundance, capturing how cells redistribute across the phenotypic space, and differential expression, identifying condition-specific transcriptional changes that may be localized, heterogeneous, or oppositely regulated across states. By modeling cell density and gene expression as continuous functions over a shared cell-state representation, Kompot enables single-cell-resolution inference with principled uncertainty estimates, without requiring predefined clusters or cell types. Applying Kompot to aging murine bone marrow, we identified a continuum of shifts in hematopoietic stem cell and mature cell states, transcriptional remodeling of monocytes independent of compositional changes, and divergent regulation of oxidative stress response genes across cell types. We demonstrate the utility of Kompot in disease settings by identifying cell-state and gene expression changes associated with improved efficacy of combinatorial immunotherapy in melanoma. Additionally, Kompot enables multi-sample comparative analysis by accounting for sample-to-sample heterogeneity. By capturing both global and cell-state-specific effects of perturbation, the Kompot framework is broadly applicable to dissecting condition-specific effects in complex single-cell landscapes.
]]></description>
<dc:creator>Otto, D. J.</dc:creator>
<dc:creator>Arriaga-Gomez, E.</dc:creator>
<dc:creator>Thieme, E.</dc:creator>
<dc:creator>Yang, R.</dc:creator>
<dc:creator>Lee, S. C.</dc:creator>
<dc:creator>Setty, M.</dc:creator>
<dc:date>2025-06-07</dc:date>
<dc:identifier>doi:10.1101/2025.06.03.657769</dc:identifier>
<dc:title><![CDATA[Comparing phenotypic manifolds with Kompot: Detecting differential abundance and gene expression at single-cell resolution]]></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/10.1101/2025.06.04.657887v1?rss=1">
<title>
<![CDATA[
Abrogating TGFβ signaling in TCR-engineered T cells and enhancing antigen processing by tumor cells promotes sustained therapeutic activity in pancreatic ductal adenocarcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.04.657887v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDA) is a deadly malignancy with limited effective therapies. Adoptive cell therapy (ACT) is a promising treatment modality for patients with solid tumors but has been limited by the highly fibroinflammatory and immunosuppressive tumor microenvironment (TME). Transforming growth factor-{beta} (TGF{beta}) participates in the inordinately suppressive TME in PDA. Here, we test the impact of selective Tgfbr2 deletion using CRISPR/Cas9 or genetic approaches in mesothelin (Msln)-specific T cell receptor (TCR) engineered T cells during ACT of PDA. Abrogating TGF{beta} signaling augmented TCR-engineered T cell accumulation in autochthonous and orthotopic PDA models and promoted terminal effector T cells, although this largely required inclusion of a vaccine at the time of T cell transfer. While loss of Tgfbr2 impaired CD103 upregulation, it only modestly impaired donor T cell central, tissue resident, or Tcf1+Slamf6+ stem-like memory T cell formation. These attributes ultimately result in heightened functional capacity and delayed tumor growth. Unexpectedly, however, most tumor-infiltrating engineered T cells failed to differentiate into PD-1+Lag3+ exhausted T cells (TEX) regardless of TGF{beta}R2 expression and despite abundant Msln protein expression by PDA cells. Forcing Msln epitope processing in KPC tumor cells promoted donor T cell accumulation, acquisition of PD-1 and Lag3, increased IFN{gamma} production by TCR-engineered T cells refractory to TGF{beta} and bypassed the vaccine requirement for therapeutic efficacy. Thus, promoting increased antigen processing/presentation by tumor cells while abrogating Tgfbr2 in engineered T cells can sustain donor T cell function in the suppressive TME and enhance the therapeutic efficacy of ACT. Our study supports pursuit of strategies that modulate tumor intrinsic antigen processing while relieving T cell suppression to safely promote the antitumor activity of TCR-engineered T cells.
]]></description>
<dc:creator>Tsai, A. K.</dc:creator>
<dc:creator>Rollins, M. R.</dc:creator>
<dc:creator>Ellefson, M. A.</dc:creator>
<dc:creator>Schmiechen, Z. C.</dc:creator>
<dc:creator>Burrack, A. L.</dc:creator>
<dc:creator>Hulbert, A.</dc:creator>
<dc:creator>Qian, G.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Provenzano, P.</dc:creator>
<dc:creator>Cruz-Hinojoza, E.</dc:creator>
<dc:creator>Butler, J. Z.</dc:creator>
<dc:creator>Smith, O. C. G.</dc:creator>
<dc:creator>O'Flanagan, S. D.</dc:creator>
<dc:creator>Krause, J.</dc:creator>
<dc:creator>Hickok, G. H.</dc:creator>
<dc:creator>Masopust, D.</dc:creator>
<dc:creator>Hingorani, S. R.</dc:creator>
<dc:creator>Greenberg, P. D.</dc:creator>
<dc:creator>Stromnes, I. M.</dc:creator>
<dc:date>2025-06-08</dc:date>
<dc:identifier>doi:10.1101/2025.06.04.657887</dc:identifier>
<dc:title><![CDATA[Abrogating TGFβ signaling in TCR-engineered T cells and enhancing antigen processing by tumor cells promotes sustained therapeutic activity in pancreatic ductal adenocarcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.09.657676v1?rss=1">
<title>
<![CDATA[
DEVELOPMENT OF A POTENT MONOCLONAL ANTIBODY FOR TREATMENT OF HUMAN METAPNEUMOVIRUS INFECTIONS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.09.657676v1?rss=1</link>
<description><![CDATA[
Human metapneumovirus (HMPV) is a major cause of respiratory infections, particularly among vulnerable populations, yet effective therapeutics remain unavailable. Monoclonal antibodies (mAbs) offer a promising approach for both treatment and prevention. Here, we describe the discovery and characterization of 4F11, a highly potent and broadly neutralizing mAb with demonstrated in vitro and in vivo efficacy against HMPV. Using cryo-electron microscopy, we defined a unique mechanism of binding HMPV employed by 4F11, which distinguishes it from previously characterized RSV and HMPV mAbs. 4F11 targets an epitope located at the apex of the prefusion F protein (site O) with a 1:1 stoichiometry, distinct from the 3:1 stoichiometry observed with other HMPV site O antibodies. Unlike other site O antibodies, which penetrate the glycan shield between Asn57 and Asn172, 4F11 binds vertically and directly interacts with the Asn172 glycan, representing a unique glycan-dependent mode of recognition. In vitro, 4F11 displayed high potency and broad neutralization across diverse HMPV strains. It also showed a low propensity for resistance development, with only a single escape mutation (K179E) identified, a mutation not found in any published HMPV sequence to date. Viruses rescued with the K179E escape mutation had significantly decreased fitness in vitro compared to wild-type virus. In a hamster challenge model, 4F11 significantly reduced viral loads in both the lungs and nasal turbinates. These findings highlight 4F11 as a promising candidate for therapeutic development, particularly for immunocompromised individuals and other high-risk groups.
]]></description>
<dc:creator>Harris, E. D.</dc:creator>
<dc:creator>McGovern, M.</dc:creator>
<dc:creator>Pernikoff, S.</dc:creator>
<dc:creator>Ikeda, R.</dc:creator>
<dc:creator>Kipnis, L.</dc:creator>
<dc:creator>Hannon, W.</dc:creator>
<dc:creator>Sobolik, E. B.</dc:creator>
<dc:creator>Gray, M.</dc:creator>
<dc:creator>Greninger, A. L.</dc:creator>
<dc:creator>He, S.</dc:creator>
<dc:creator>Chin, C.-N.</dc:creator>
<dc:creator>Fu, T.-M.</dc:creator>
<dc:creator>Pancera, M.</dc:creator>
<dc:creator>Boonyaratanakornkit, J.</dc:creator>
<dc:date>2025-06-11</dc:date>
<dc:identifier>doi:10.1101/2025.06.09.657676</dc:identifier>
<dc:title><![CDATA[DEVELOPMENT OF A POTENT MONOCLONAL ANTIBODY FOR TREATMENT OF HUMAN METAPNEUMOVIRUS INFECTIONS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.10.657742v1?rss=1">
<title>
<![CDATA[
Biological causes and impacts of rugged tree landscapes in phylodynamic inference 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.10.657742v1?rss=1</link>
<description><![CDATA[
Phylodynamic analysis has been instrumental in elucidating epidemiological and evolutionary dynamics of pathogens. Bayesian phylodynamics integrates out phylogenetic uncertainty, which is typically substantial in phylodynamic datasets due to limited genetic diversity. Phylodynamic inference does not, however, scale with modern datasets, partly due to difficulties in traversing tree space. Here, we characterize tree space and landscape in phylodynamic inference and assess its impacts on analysis difficulty and key biological estimates. By running extensive Bayesian analyses of 15 classic large phylodynamic datasets and carefully analyzing the posterior samples, we find that the posterior tree landscape is diffuse yet rugged, leading to widespread tree sampling problems that usually stem from sequences in a small part of the tree. We develop clade-specific diagnostics to show that a few sequences--including putative recombinants and recurrent mutants--frequently drive the ruggedness and sampling problems, although existing data-quality tests show limited power to detect them. The sampling problems can significantly impact phylodynamic inferences or distort major biological conclusions; the impact is usually stronger on "local" estimates (e.g., introduction history) associated with particular clades than on "global" parameters (e.g., demographic trajectory) governed by general tree shape. We evaluate existing and newly-developed MCMC diagnostics, and offer strategies for optimizing phylodynamic analysis settings and mitigating sampling problem impacts. Our findings highlight the need and directions to develop efficient traversal over rugged tree landscapes, ultimately advancing scalable and reliable phylodynamics.

Significance StatementBayesian phylodynamics is central to epidemiological studies, but exploring the vast and complex tree space is computationally challenging. Phylodynamic datasets comprise many highly similar sequences, sampled through time, creating a uniquely structured landscape of optimal trees. Here, we show that phylodynamic tree landscapes are often highly rugged, with multiple peaks separated by difficult-to-cross valleys. These features lead to widespread sampling problems which are often driven by a few sequences. These problems can significantly impact phylodynamic estimates, especially those associated with particular clades, distorting biological conclusions. We develop diagnostics to identify problematic sequences and provide solutions to mitigate their impacts. We offer strategies to optimize phylodynamic analysis workflows and to develop algorithms for navigating rugged landscapes, thereby advancing infectious disease investigation.
]]></description>
<dc:creator>Gao, J.</dc:creator>
<dc:creator>Brusselmans, M.</dc:creator>
<dc:creator>Carvalho, L. M.</dc:creator>
<dc:creator>Suchard, M. A.</dc:creator>
<dc:creator>Baele, G.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2025-06-12</dc:date>
<dc:identifier>doi:10.1101/2025.06.10.657742</dc:identifier>
<dc:title><![CDATA[Biological causes and impacts of rugged tree landscapes in phylodynamic inference]]></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/10.1101/2025.06.12.659400v1?rss=1">
<title>
<![CDATA[
Seasonal influenza viruses show distinct adaptive dynamics during growth in chicken eggs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.12.659400v1?rss=1</link>
<description><![CDATA[
Human influenza viruses are grown in chicken eggs for vaccine production. Sequences of these egg-passaged viruses give us the opportunity to examine the evolution that occurs when these human viruses are subjected to the selective pressure of growing in chicken eggs, which (among other things) express different sialic acid receptors. The repetition of this evolutionary experiment in hundreds of strains over the past several decades allows us to identify mutations that adapt the virus to eggs and epistatic constraints that influence them. We analyze influenza A/H3N2, A/H1N1pdm, B/Vic and B/Yam sequences that were passaged in eggs and find that almost all of the adaptive mutations are located around the receptor-binding pocket of hemagglutinin (HA). We observe epistatic interactions both between adaptive mutations and between these mutations and the continually-evolving human influenza HA background sequence. Our results show that this background dependence is greatest for influenza A/H3N2, then A/H1N1pdm, with B/Vic and B/Yam showing little-to-no background dependence. We find that the total number of adaptive mutations and the length of adaptive walk also follow the same pattern between the influenza subtypes, suggesting that background dependence, number of adaptive mutations, and extent of additive versus epistatic interactions may all be related features of the fitness landscape.
]]></description>
<dc:creator>Kistler, K.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.12.659400</dc:identifier>
<dc:title><![CDATA[Seasonal influenza viruses show distinct adaptive dynamics during growth in chicken eggs]]></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/10.1101/2025.06.16.659977v1?rss=1">
<title>
<![CDATA[
Nucleotide context models outperform protein language models for predicting antibody affinity maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.16.659977v1?rss=1</link>
<description><![CDATA[
Antibodies play a crucial role in adaptive immunity. They develop as B cell receptors (BCRs): membrane-bound forms of antibodies that are expressed on the surfaces of B cells. BCRs are refined through affinity maturation, a process of somatic hypermutation (SHM) and natural selection, to improve binding to an antigen. Computational models of affinity maturation have developed from two main perspectives: molecular evolution and language modeling. The molecular evolution perspective focuses on nucleotide sequence context to describe mutation and selection; the language modeling perspective involves learning patterns from large data sets of protein sequences. In this paper, we compared models from both perspectives on their ability to predict the course of antibody affinity maturation along phylogenetic trees of BCR sequences. This included models of SHM, models of SHM combined with an estimate of selection, and protein language models. We evaluated these models for large human BCR repertoire data sets, as well as an antigen-specific mouse experiment with a pre-rearranged cognate naive antibody. We demonstrated that precise modeling of SHM, which requires the nucleotide context, provides a substantial amount of predictive power for predicting the course of affinity maturation. Notably, a simple nucleotide-based convolutional neural network modeling SHM outperformed state-of-the-art protein language models, including one trained exclusively on antibody sequences. Furthermore, incorporating estimates of selection based on a custom deep mutational scanning experiment brought only modest improvement in predictive power. To support further research, we introduce EPAM (Evaluating Predictions of Affinity Maturation), a benchmarking framework to integrate evolutionary principles with advances in language modeling, offering a road map for understanding antibody evolution and improving predictive models.
]]></description>
<dc:creator>Johnson, M. M.</dc:creator>
<dc:creator>Sung, K.</dc:creator>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Vora, A. A.</dc:creator>
<dc:creator>Araki, T.</dc:creator>
<dc:creator>Victora, G. D.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Fukuyama, J.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2025-06-18</dc:date>
<dc:identifier>doi:10.1101/2025.06.16.659977</dc:identifier>
<dc:title><![CDATA[Nucleotide context models outperform protein language models for predicting antibody affinity maturation]]></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/10.1101/2025.06.16.660016v1?rss=1">
<title>
<![CDATA[
Learning Human T Cell Behaviors through Generative AI Embeddings of T Cell Receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.16.660016v1?rss=1</link>
<description><![CDATA[
T cells interact with the world through T cell receptors (TCRs). The extent to which TCRs determine T cell behavior has not been comprehensively characterized. Our Tarpon model leverages advances in generative artificial intelligence to synthesize large-scale (>1M sequences) TCR atlases across human development and diseases into actionable insights. Tarpon creates: 1) bespoke sampling functions generating realistic Ag-specific TCRs, 2) embeddings revealing CD4+ and CD8+ single-positive TCR repertoires as distinct with divergent physiochemical properties, and 3) cross-dataset mappings of T cell states that validate fetal CD4+ versus CD8+ TCR differences in adults and find fetal type I innate T cells to map to MAIT and KIR+ adult CD8+ T cells which we verify via whole transcriptome analysis. Tarpon is a resource as a reference of TCRs across human physiological states and as a computational framework to create interpretable TCR embeddings, via physicochemical associations, that have broad implications for the field.
]]></description>
<dc:creator>Chen, D. G.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Heath, J. R.</dc:creator>
<dc:date>2025-06-22</dc:date>
<dc:identifier>doi:10.1101/2025.06.16.660016</dc:identifier>
<dc:title><![CDATA[Learning Human T Cell Behaviors through Generative AI Embeddings of T Cell Receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.18.660465v1?rss=1">
<title>
<![CDATA[
Development of PROTACs for targeted degradation of oncogenic TRK fusions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.18.660465v1?rss=1</link>
<description><![CDATA[
Chromosomal translocations leading to the fusion of tropomyosin receptor kinases (TRK) with diverse partner proteins have been identified as oncogenic drivers in many adult and pediatric cancers. While first-generation TRK kinase inhibitors, such as entrectinib and larotrectinib, have shown positive responses in TRK fusion-positive cancers, resistance mutations against these inhibitors in the kinase domain limit their efficacy. Second-generation inhibitors are in clinical evaluation, highlighting a need for novel therapeutic modalities to achieve durable suppression of the oncogenic activity of TRK fusions. Here, we developed heterobifunctional small molecule degraders (PROTACs) to achieve targeted degradation of TRK fusions. By conjugating entrectinib to thalidomide, we identified JWJ-01-378 as a potent and selective CRBN-recruiting degrader of the TPM3-TRKA fusion. JWJ-01-378 induced TPM3-TRKA degradation through the ubiquitin-proteasome system and proteomics analysis confirmed the acute selectivity of JWJ-01-378 for achieving TPM3-TRKA degradation with minimal off-target effects. While JWJ-01-378 was also able to degrade wild-type TRK, it was unable to degrade TRK inhibitor resistant mutants and ALK fusions. Importantly, TPM3-TRKA degradation by JWJ-01-378 suppressed downstream signaling and reduced cancer cell viability, with improved responses compared to heterobifunctional control compounds that cannot degrade TPM3-TRKA. Together, our study expands the toolbox of compounds for evaluating targeted degradation of TRK fusions in cancer.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=97 SRC="FIGDIR/small/660465v1_ufig13.gif" ALT="Figure 13">
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org.highwire.dtl.DTLVardef@6c78f7org.highwire.dtl.DTLVardef@179dcdeorg.highwire.dtl.DTLVardef@1939988org.highwire.dtl.DTLVardef@144c718_HPS_FORMAT_FIGEXP  M_FIG C_FIG JWJ-01-378 recruits cereblon (CRBN) to induce potent and selective degradation of oncogenic TRK fusions, leading to a collapse in downstream signaling and loss of cancer cell viability. Graphical abstract was created using Biorender.com.
]]></description>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Jiang, J.</dc:creator>
<dc:creator>Donald-Paladino, M. S.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Gutierrez, A.</dc:creator>
<dc:creator>Federation, A. J.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:creator>Holland, E. C.</dc:creator>
<dc:creator>Ferguson, F. M.</dc:creator>
<dc:creator>Nabet, B.</dc:creator>
<dc:date>2025-06-24</dc:date>
<dc:identifier>doi:10.1101/2025.06.18.660465</dc:identifier>
<dc:title><![CDATA[Development of PROTACs for targeted degradation of oncogenic TRK fusions]]></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/10.1101/2025.06.19.660576v1?rss=1">
<title>
<![CDATA[
Obesity promotes conserved inflammatory and metabolic transcriptional programs in mouse and human colon tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.19.660576v1?rss=1</link>
<description><![CDATA[
BackgroundThe global prevalence of obesity, an established risk and progression factor for colon cancer, is high and rising. Unfortunately, the mechanisms underlying the obesity-colon cancer association are incompletely understood, and new molecular targets enabling more effective intervention strategies to break the obesity-colon cancer link are urgently needed.

ObjectiveThis study integrated RNA sequencing data from mouse and human colon tumor samples, as well as human adipose samples, to rigorously establish obesity-associated transcriptomic signatures conserved between the two species.

MethodsWe employed a mouse colon cancer model with colonoscopy-guided orthotopic transplantation of syngeneic Apc-null;KrasG12D/+;Trp53-null;Smad4-null;tdTomato colon tumor organoids. Epithelial cell adhesion molecule (EpCAM)-positive cells from murine tumors, and 193 human colon tumors and 188 human mesenteric adipose tissue samples from the ColoCare cohort underwent transcriptomic analyses.

ResultsDiet-induced obesity reduced survival in the mouse model of colon cancer. Integrated transcriptomic analyses of EpCAM-positive murine tumor cells and bulk human tumors revealed obesity-driven enrichment of inflammation and metabolic pathways, including upregulation of genes involved in innate immune sensing (TLR2, MYD88, IRF4) and tumor microenvironment remodeling (MMP9, TGFB1, SERPINE1). Analysis of paired mesenteric visceral adipose tissue and tumor samples from the ColoCare cohort indicated that obesity amplifies inflammatory signaling pathways through unique adipose ligand-tumor receptor interactions.

ConclusionsThese results establish obesity-associated adipose tissue dysregulation as a key inter-tissue modulator of biology, with concordant cross-species effects on tumor cell-intrinsic inflammatory and metabolic programs.
]]></description>
<dc:creator>Glenny, E. M.</dc:creator>
<dc:creator>Lin, T.</dc:creator>
<dc:creator>Bandera, V. M.</dc:creator>
<dc:creator>Mirminachi, B.</dc:creator>
<dc:creator>Khumukcham, S. S.</dc:creator>
<dc:creator>Gigic, B.</dc:creator>
<dc:creator>Warby, C. A.</dc:creator>
<dc:creator>Aksonova, O.</dc:creator>
<dc:creator>Coleman, M. F.</dc:creator>
<dc:creator>Carpanese, A.</dc:creator>
<dc:creator>Busch, C.</dc:creator>
<dc:creator>Himbert, C.</dc:creator>
<dc:creator>Ose, J.</dc:creator>
<dc:creator>Nix, D. A.</dc:creator>
<dc:creator>Boucher, K.</dc:creator>
<dc:creator>Schirmacher, P.</dc:creator>
<dc:creator>Strehli, I.</dc:creator>
<dc:creator>Hardikar, S.</dc:creator>
<dc:creator>Cohan, J. N.</dc:creator>
<dc:creator>Jedrzkiewicz, J.</dc:creator>
<dc:creator>Brobeil, A.</dc:creator>
<dc:creator>Schneider, M. A.</dc:creator>
<dc:creator>Kahlert, C.</dc:creator>
<dc:creator>Siegel, E. M.</dc:creator>
<dc:creator>Byrd, D. A.</dc:creator>
<dc:creator>Toriola, A. T.</dc:creator>
<dc:creator>Shibata, D.</dc:creator>
<dc:creator>Li, C. I.</dc:creator>
<dc:creator>Figueiredo, J. C.</dc:creator>
<dc:creator>Tan, A. C.</dc:creator>
<dc:creator>Roper, J.</dc:creator>
<dc:creator>Ulrich, C. M.</dc:creator>
<dc:creator>Hursting, S. D.</dc:creator>
<dc:date>2025-06-25</dc:date>
<dc:identifier>doi:10.1101/2025.06.19.660576</dc:identifier>
<dc:title><![CDATA[Obesity promotes conserved inflammatory and metabolic transcriptional programs in mouse and human colon tumors]]></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/10.1101/2025.06.25.661543v1?rss=1">
<title>
<![CDATA[
Herpes simplex virus 1 strain 17+ with R2 mutation in UL37 has residual retrograde transport 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.25.661543v1?rss=1</link>
<description><![CDATA[
Herpes simplex virus 1 (HSV-1) causes lifelong recurrent infections. Following primary infection of the oral or genital mucosa, HSV-1 travels retrogradely through axons and establishes latency in the cell body of ganglionic neurons of the peripheral nervous system. Periodic reactivation in neurons and anterograde transport of virions back to peripheral regions causes oral or genital ulcerations. Many host and viral factors implicated in retrograde and anterograde transport of HSV-1 have been identified. In particular, studies reported that introducing five amino acid substitutions in the R2 region of the viral tegument protein UL37 was sufficient to completely eliminate retrograde transport of HSV-1 strain F. Here, we introduced the same R2 mutations in the highly neurovirulent HSV-1 strain 17+. We report that this R217 virus has residual retrograde travel. We show that R217 can establish latency in mouse models of ocular and vaginal infection and reactivate. These results contradict published evidence and show that the R2 mutation is not sufficient to fully prevent retrograde transport of HSV-1.
]]></description>
<dc:creator>Walter, M.</dc:creator>
<dc:creator>Haick, A. K.</dc:creator>
<dc:creator>Massa, P. A.</dc:creator>
<dc:creator>Klouser, L. M.</dc:creator>
<dc:creator>Stensland, L.</dc:creator>
<dc:creator>Santo, T. K.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:date>2025-06-26</dc:date>
<dc:identifier>doi:10.1101/2025.06.25.661543</dc:identifier>
<dc:title><![CDATA[Herpes simplex virus 1 strain 17+ with R2 mutation in UL37 has residual retrograde transport]]></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/10.1101/2025.06.25.661565v1?rss=1">
<title>
<![CDATA[
Direct observation of interdependent and hierarchical kinetochore assembly on individual centromeres 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.25.661565v1?rss=1</link>
<description><![CDATA[
Kinetochores are megadalton protein machines that harness microtubules to segregate chromosomes during cell division. The kinetochores must assemble after DNA replication during every cell cycle onto specialized regions of chromosomes called centromeres, but the order and regulation of their assembly remains unclear due to the complexity of kinetochore composition and the difficulty resolving individual kinetochores in vivo. Here, by adapting a prior single-molecule method for monitoring kinetochore assembly in budding yeast lysates, we identify a sequential order of assembly and uncover previously unknown interdependencies between subcomplexes. We show that inner kinetochore assembly depends partly on outer kinetochore components, and that outer kinetochore branches do not assemble independently of one another. Notably, Mif2 assembly is a rate-limiting step that can be accelerated by binding to the Mtw1 subcomplex, thereby promoting rapid assembly of many inner and outer kinetochore components. The importance of controlling kinetochore assembly kinetics is supported by a Mif2 mutant lacking both autoinhibition and Mtw1 subcomplex binding activity, which leads to defective kinetochore-microtubule attachments when the centromeric histone variant Cse4 is overexpressed. Altogether, our work provides a direct view of kinetochore assembly and reveals highly interdependent regulatory events that control its order and timing.

GRAPHICAL ABSTRACT

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=64 SRC="FIGDIR/small/661565v1_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@bcc252org.highwire.dtl.DTLVardef@79b6b2org.highwire.dtl.DTLVardef@c974e0org.highwire.dtl.DTLVardef@184689_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Hu, C.</dc:creator>
<dc:creator>Popchock, A. R.</dc:creator>
<dc:creator>Latino, A. A.</dc:creator>
<dc:creator>Asbury, C. L.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2025-06-26</dc:date>
<dc:identifier>doi:10.1101/2025.06.25.661565</dc:identifier>
<dc:title><![CDATA[Direct observation of interdependent and hierarchical kinetochore assembly on individual centromeres]]></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/10.1101/2025.06.24.661157v1?rss=1">
<title>
<![CDATA[
Genital herpes shedding episodes associate with alterations in the spatial organization and activation of mucosal immune cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.24.661157v1?rss=1</link>
<description><![CDATA[
Herpes Simplex Virus 2 (HSV-2) infection results in variable rates of local viral shedding in anogenital skin. The impact of episodic viral exposures on immune cells in adjacent mucosal tissues, including the genital tract, is unknown. However, any immune responses at this site could impact protective mucosal immunity, tissue homeostasis, and adverse health outcomes. To investigate the impact of HSV-2 on cervicovaginal tract immunity, we applied flow cytometry, immunofluorescent imaging, analysis of soluble immune factors, and spatial transcriptomics to cervicovaginal tissue and blood samples provided by a total of 232 HSV-2 seropositive and seronegative participants, with genital HSV-2 shedding evaluated at the time of biopsy. This unique dataset was used to define and spatially map immune cell subsets and localized gene expression via spatial transcriptomics. HSV-2 seropositivity alone was associated with minimal differences in cervicovaginal and circulating T cell phenotypes. However, the vaginal mucosa during active HSV-2 shedding was associated with alterations in T cell, macrophage, and dendritic cell localization and gene expression consistent with increased immune surveillance, with immune activating and suppressing signals potentially reinforcing mucosal tissue homeostasis.

SummaryIn context of episodic HSV-2 shedding, immune cells mobilize and co-localize in the vaginal epithelium, expressing cytotoxic and inflammatory genes and immunoregulatory genes that collectively may promote tissue homeostasis in settings of episodic viral shedding to limit damage.
]]></description>
<dc:creator>MacLean, F.</dc:creator>
<dc:creator>Zemek, R. M.</dc:creator>
<dc:creator>Tsegaye, A. T.</dc:creator>
<dc:creator>Graham, J. B.</dc:creator>
<dc:creator>Swarts, J. L.</dc:creator>
<dc:creator>Vick, S. C.</dc:creator>
<dc:creator>Potchen, N. B.</dc:creator>
<dc:creator>Talavera, I. C.</dc:creator>
<dc:creator>Warrier, L.</dc:creator>
<dc:creator>Dubrulle, J.</dc:creator>
<dc:creator>Schroeder, L. K.</dc:creator>
<dc:creator>Elz, A.</dc:creator>
<dc:creator>Sowerby, D.</dc:creator>
<dc:creator>Saito, A.</dc:creator>
<dc:creator>Thomas, K. K.</dc:creator>
<dc:creator>Mack, M.</dc:creator>
<dc:creator>Schiffer, J.</dc:creator>
<dc:creator>McClelland, R. S.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Chohan, B. H.</dc:creator>
<dc:creator>Ngure, K.</dc:creator>
<dc:creator>Mugo, N. R.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Lingappa, J. R.</dc:creator>
<dc:creator>Lund, J. M.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.661157</dc:identifier>
<dc:title><![CDATA[Genital herpes shedding episodes associate with alterations in the spatial organization and activation of mucosal immune cells]]></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/10.1101/2025.07.01.661217v1?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">
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]]></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/10.1101/2025.07.02.662848v1?rss=1">
<title>
<![CDATA[
PDL1 CHECKPOINT BLOCKADE SYNERGIZES WITH NILOTINIB BUT NOT DASATINIB TO PREVENT LEUKEMIA RELAPSE 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.02.662848v1?rss=1</link>
<description><![CDATA[
Dasatinib and nilotinib are front line Tyrosine Kinase Inhibitors (TKIs) used to treat BCR-ABL+ B cell acute lymphoblastic leukemia (B-ALL) and BCR-ABL+ chronic myelogenous leukemia. We previously showed that combining nilotinib with anti-PD-L1 blockade significantly reduced leukemia relapse. The TKI dasatinib is more commonly used to treat to B-ALL. However, unlike nilotinib, dasatinib also inhibits SRC-family kinases, which may make it less efficacious in combination with anti-PDL1 blockade. Herein we assess the impact of nilotinib versus dasatinib on anti-leukemia immune responses. Dasatinib, but not nilotinib, inhibited T cell proliferation at high doses in-vitro, but neither TKI significantly impacted T cell function or expansion in response to immunization in-vivo with a model antigen (2W1S) plus polyIC-adjuvant. Dasatinib and nilotinib both reduced leukemic blasts equivalently after 5 days of treatment. In contrast, nilotinib and PD-L1 blockade, but not dasatinib plus anti-PDL1, prevented relapse several weeks later. Thus, dasatinib negatively impacts protective anti-leukemia T cell responses that prevent leukemia relapse.
]]></description>
<dc:creator>Morgan, E. C.</dc:creator>
<dc:creator>Venkatesh, H.</dc:creator>
<dc:creator>Centenos, E. G.</dc:creator>
<dc:creator>Heltemes-Harris, L. M.</dc:creator>
<dc:creator>Hubbard, G.</dc:creator>
<dc:creator>Maiers, T. L.</dc:creator>
<dc:creator>Dileepan, T.</dc:creator>
<dc:creator>Tracy, S. I.</dc:creator>
<dc:creator>Farrar, M. A.</dc:creator>
<dc:date>2025-07-05</dc:date>
<dc:identifier>doi:10.1101/2025.07.02.662848</dc:identifier>
<dc:title><![CDATA[PDL1 CHECKPOINT BLOCKADE SYNERGIZES WITH NILOTINIB BUT NOT DASATINIB TO PREVENT LEUKEMIA RELAPSE]]></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/10.1101/2025.07.01.659979v1?rss=1">
<title>
<![CDATA[
Influence of the broadly neutralizing antibody VRC01 on HIV breakthrough virus populations in antibody-mediated prevention trials 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.01.659979v1?rss=1</link>
<description><![CDATA[
In the HIV antibody mediated prevention (AMP) trials, the broadly neutralizing antibody VRC01 demonstrated protective efficacy against new diagnoses with susceptible HIV strains. To understand how VRC01 shaped breakthrough infections, we performed deep sequencing on 172 participants in the placebo and treatment arms, generating 63,444 gag-{Delta}pol (2.5 kb) and 53,088 rev-env-{Delta}nef (3 kb) sequences. Sequences were classified into transmitted founder lineages (TFLs), and infections with multiple distinct lineages were determined. Multilineage infections were detected in [~]38% of participants in both the African (HVTN 703/HPTN 081) and Americas/Europe (HVTN 704/HPTN 085) cohorts, regardless of placebo or treatment group, or cohort. The high levels of multilineage infections could be attributed to minor lineages (<5% abundance) identified in 20% of participants. Infection with VRC01 discordant viruses (IC80s >3-fold different) was observed in 40% of multilineage infections, with a trend toward greater intra-host neutralization differences with increasing VRC01 dose (Jonckheere-Terpstra test, p=0.072). In six VRC01 treated participants who acquired both sensitive (IC80<1{micro}g/ml) and resistant viruses (IC80>3{micro}g/ml), the sensitive lineages declined over time. Recombination was pervasive, observed in 63% of multilineage infections at the time of HIV diagnosis. In one treated participant infected with VRC01 discordant lineages, recombinant viruses preferentially inherited the resistance mutation (binomial p=0.004). In conclusion, our in-depth analysis of breakthrough viruses in the AMP trials revealed a high frequency of multilineage infections, including infections with viruses with different VRC01 sensitivities. This analysis also highlights the role of recombination in shaping intra-host viral evolution and facilitating escape from VRC01.

SignificanceThis work advances our understanding of the diversity of initial viral infection and evolution and in vivo activity of broadly neutralizing antibodies (bNAbs). Deep sequencing revealed [~]38% of HIV acquisitions with multiple transmitted founder lineages, higher than previous studies. These occurrences were similar in the placebo and VRC01 groups. Viral recombination among post-acquisition variants was common under antibody selection and appeared to favor resistant sequences in the treatment group. These data suggest as with single antiviral therapy, passive and active immunization of bNAbs should be directed at multiple antigen targets of HIV-1.
]]></description>
<dc:creator>Williamson, C.</dc:creator>
<dc:creator>Moodley, C.</dc:creator>
<dc:creator>Magaret, C. A.</dc:creator>
<dc:creator>Giorgi, E. E.</dc:creator>
<dc:creator>Rolland, M.</dc:creator>
<dc:creator>Westfall, D. H.</dc:creator>
<dc:creator>Yssel, A.</dc:creator>
<dc:creator>Deng, W.</dc:creator>
<dc:creator>Rossenkhan, R.</dc:creator>
<dc:creator>Mkhize, N. N.</dc:creator>
<dc:creator>Chen, L. H.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Bhattacharya, T.</dc:creator>
<dc:creator>Pankow, A.</dc:creator>
<dc:creator>Murrell, B.</dc:creator>
<dc:creator>York, T.</dc:creator>
<dc:creator>Gwashu-Nyangiwe, A.</dc:creator>
<dc:creator>Ndabambi, N.</dc:creator>
<dc:creator>Thebus, R.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Lambson, B. E.</dc:creator>
<dc:creator>Kaldine, H.</dc:creator>
<dc:creator>Bhebhe, S.</dc:creator>
<dc:creator>Juraska, M.</dc:creator>
<dc:creator>Bai, H.</dc:creator>
<dc:creator>deCamp, A. C.</dc:creator>
<dc:creator>Miner, M.</dc:creator>
<dc:creator>Ludwig, J.</dc:creator>
<dc:creator>Molitor, C.</dc:creator>
<dc:creator>Beaume, N.</dc:creator>
<dc:creator>Matten, D.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Reeves, D. B.</dc:creator>
<dc:creator>Mayer, B.</dc:creator>
<dc:creator>Karuna, S. T.</dc:creator>
<dc:creator>Hural, J. A.</dc:creator>
<dc:creator>Morris, L.</dc:creator>
<dc:creator>Montefiori, D.</dc:creator>
<dc:creator>Bumgarner, R. E.</dc:creator>
<dc:creator>Moore, P.</dc:creator>
<dc:creator>Edlefsen, P. T.</dc:creator>
<dc:creator>Edupuganti, S.</dc:creator>
<dc:creator>Mgodi, N.</dc:creator>
<dc:creator>McElrath</dc:creator>
<dc:date>2025-07-10</dc:date>
<dc:identifier>doi:10.1101/2025.07.01.659979</dc:identifier>
<dc:title><![CDATA[Influence of the broadly neutralizing antibody VRC01 on HIV breakthrough virus populations in antibody-mediated prevention trials]]></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/10.1101/2025.07.11.664404v1?rss=1">
<title>
<![CDATA[
A Singular Base Editing Platform for Polyfunctional Multiplex Engineering of Immune Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.11.664404v1?rss=1</link>
<description><![CDATA[
Current methods to engineer antigen-specific receptors rely on randomly integrating vectors or double-strand break induced targeted integration, both of which pose safety risks. To implement an all-in-one tool for multiplex knockout (KO) and knock in (KI), we expand the use of cytosine and adenine base editor (ABE) nickase activity to stimulate homology-directed repair (HDR) and insert clinically relevant chimeric antigen receptors (CARs) into specific loci. Through a novel sgRNA design strategy and a recombinant adeno-associated virus (rAAV) delivered DNA template, we enhanced the efficiency of ABE8e-stimulated HDR in human T cells. By combining KI of CD19-, CD33-, or mesothelin-targeting CARs with >95% quadplex gene KO (B2M/CD3{varepsilon}/PDCD1/CISH), we achieve single-step generation of highly functional off-the-shelf CAR T cell products with enhanced function. Importantly, we found no detectable translocations or significant off-target edits and demonstrated efficacy against multiple cancer lines, and a suppressive 3D spheroid culture model. This efficient engineering process of Iterative Nicking for Synchronous Engineered Reprogramming of T cells (INSERT) establishes a safe, simplified platform for advanced therapeutic CAR T engineering.
]]></description>
<dc:creator>Skeate, J. G.</dc:creator>
<dc:creator>Slipek, N. J.</dc:creator>
<dc:creator>Lahr, W. S.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:creator>Wick, B. J.</dc:creator>
<dc:creator>Stelljes, E. M.</dc:creator>
<dc:creator>Gilkey, A. K.</dc:creator>
<dc:creator>Thenge, P. P.</dc:creator>
<dc:creator>Diers, M. D.</dc:creator>
<dc:creator>Kar, B.</dc:creator>
<dc:creator>Krueger, J. B.</dc:creator>
<dc:creator>Niemeyer, E. M.</dc:creator>
<dc:creator>Lonetree, C.-l.</dc:creator>
<dc:creator>Kluesner, M. G.</dc:creator>
<dc:creator>Bell, J. B.</dc:creator>
<dc:creator>Clement, K.</dc:creator>
<dc:creator>Provenzano, P.</dc:creator>
<dc:creator>Moriarity, B. S.</dc:creator>
<dc:creator>Webber, B. R.</dc:creator>
<dc:date>2025-07-16</dc:date>
<dc:identifier>doi:10.1101/2025.07.11.664404</dc:identifier>
<dc:title><![CDATA[A Singular Base Editing Platform for Polyfunctional Multiplex Engineering of Immune Cells]]></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/10.1101/2025.07.14.664835v1?rss=1">
<title>
<![CDATA[
Humanized mice enable in vivo evaluation of engineered plasma cell biology and therapeutic function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.14.664835v1?rss=1</link>
<description><![CDATA[
Engineered plasma cells (ePCs) offer a durable strategy for in vivo delivery of therapeutic antibodies, but standard immunodeficient mouse models lack human immune factors critical for plasma cell survival and function. We utilized a humanized mouse model in which NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice were engrafted with human CD34+ stem cells as recipients for infusions with autologous ePCs. In this setting, ePCs localized to plasma cell niches and stably secreted antibodies for over three months. To improve the selection of antibodies for secretion, we developed a B cell receptor surface display screen that identified candidate antibody sequences with high secretion potential. An anti-SARS-CoV-2 antibody (clone 297) selected by this method showed robust secretion both in vitro and vivo, and serum from ePC-engrafted mice potently neutralized SARS-CoV-2 pseudovirus. Together, these findings establish a physiologically relevant model for testing human ePCs, and offer a generalizable strategy for optimizing antibody selection to support long-term therapeutic delivery.
]]></description>
<dc:creator>Hill, T. F.</dc:creator>
<dc:creator>Helmers, A. E.</dc:creator>
<dc:creator>Cheng, R. Y.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Homad, L. J.</dc:creator>
<dc:creator>Narvekar, P.</dc:creator>
<dc:creator>Asher, G. D.</dc:creator>
<dc:creator>Camp, N. D.</dc:creator>
<dc:creator>Suchland, E. R.</dc:creator>
<dc:creator>Ott, A. R.</dc:creator>
<dc:creator>Hale, M.</dc:creator>
<dc:creator>Thouvenel, C. D.</dc:creator>
<dc:creator>Stoffers, C.</dc:creator>
<dc:creator>Lachkar, S.</dc:creator>
<dc:creator>McGuire, A.</dc:creator>
<dc:creator>Rawlings, D. J.</dc:creator>
<dc:creator>James, R. G.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.14.664835</dc:identifier>
<dc:title><![CDATA[Humanized mice enable in vivo evaluation of engineered plasma cell biology and therapeutic function]]></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/10.1101/2025.07.18.665436v1?rss=1">
<title>
<![CDATA[
TCR2HLA: calibrated inference of HLA genotypes from TCR repertoires enables identification of immunologically relevant metaclonotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.18.665436v1?rss=1</link>
<description><![CDATA[
T cell receptors (TCRs) recognize peptides presented by polymorphic human leukocyte antigen (HLA) molecules, but HLA genotype data are often missing from TCR repertoire sequencing studies. To address this, we developed TCR2HLA, an open-source tool that infers HLA genotypes from TCR{beta} repertoires. Expanding on work linking public TRBV-CDR3 sequences to HLA genotypes, we incorporated "quasi-public" metaclonotypes - composed of rarer TCR{beta} sequences with shared amino acid features - enriched by HLA genotypes. Using four TCR{beta}seq datasets from 3,150 individuals, we applied TRBV gene partitioning and locality-sensitive hashing to identify [~]96,000 TCR{beta} features strongly associated with specific HLA alleles from 71M input TCRs. Binary HLA classifiers built with these features achieved high balanced accuracy (>0.9) across common HLA-A (9/12), B (9/12), C (6/13), DRB1 (11/11) alleles and prevalent DPA1/DPB1 (6/10), DQA1/DQB1 (8/17) heterodimers. We also introduced a high-sensitivity calibration to support predictions in samples with as few as 5,000 unique clonotypes. Calibrated predictions with confidence filtering improved reliability. Beyond genotype imputation, TCR2HLA enables the discovery of novel HLA- and exposure-associated TCRs, as shown by the identification of SARS-CoV-2 related TCRs in a large COVID-19 dataset lacking HLA data. TCR2HLA provides a scalable framework for bridging the gap between TCRseq data and HLA genotype for biomarker discovery.
]]></description>
<dc:creator>Mayer-Blackwell, K.</dc:creator>
<dc:creator>Minervina, A.</dc:creator>
<dc:creator>Pogorelyy, M.</dc:creator>
<dc:creator>Rawat, P.</dc:creator>
<dc:creator>Shapiro, M. R.</dc:creator>
<dc:creator>Peters, L. D.</dc:creator>
<dc:creator>Ford, E. S.</dc:creator>
<dc:creator>Posgai, A. L.</dc:creator>
<dc:creator>Vengesana, K.</dc:creator>
<dc:creator>Minot, S.</dc:creator>
<dc:creator>Koelle, D. M.</dc:creator>
<dc:creator>Greiff, V.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Brusko, T. M.</dc:creator>
<dc:creator>Thomas, P. G.</dc:creator>
<dc:creator>Fiore-Gartland, A.</dc:creator>
<dc:date>2025-07-23</dc:date>
<dc:identifier>doi:10.1101/2025.07.18.665436</dc:identifier>
<dc:title><![CDATA[TCR2HLA: calibrated inference of HLA genotypes from TCR repertoires enables identification of immunologically relevant metaclonotypes]]></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/10.1101/2025.07.22.666002v1?rss=1">
<title>
<![CDATA[
B cells specific for polyomavirus-derived oncoprotein are predictive of Merkel cell carcinoma progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.22.666002v1?rss=1</link>
<description><![CDATA[
Merkel cell carcinomas typically arise from clonal integration of the Merkel cell polyomavirus. Immunogenic viral oncoproteins then lead to tumorigenesis. Oncoprotein-specific T cells are essential for anti-MCC immunity, but it is unclear whether B cells promote tumor control. Here, we analyzed the frequency and phenotype of viral oncoprotein-specific and total B cells in 47 blood samples and 19 unmatched tumors from MCC patients-- of which 8 out 19 progressed. The phenotype of blood B cells did not correlate with MCC patient outcomes. In contrast, all 11 patients with robust oncoprotein-specific antibody-secreting and/or germinal center B cells in tumors experienced long-term MCC control. In vitro, B cells engineered to be specific for viral oncoproteins increased the sensitivity of oncoprotein-specific CD4+ T cells by over 50-fold. Together, our findings suggest that cancer-specific B cells promote anti-tumor immunity via increased T cell responses and that cancer-specific B cell augmentation could be therapeutically relevant.

Statement of SignificanceThe link between cancer-specific B cells in anti-tumor immunity and clinical outcomes remains poorly defined. Here, we show that tumor-associated B cells specific for a viral oncoprotein expressed in MCC patient tumors predict disease control with remarkable accuracy, establishing their potential as active participants in tumor immunity.
]]></description>
<dc:creator>Rodriguez Chevez, H. J.</dc:creator>
<dc:creator>Remington, A. J.</dc:creator>
<dc:creator>Gray, M. M.</dc:creator>
<dc:creator>Alam, R.</dc:creator>
<dc:creator>Gilmour, M. W.</dc:creator>
<dc:creator>Morningstar, C.</dc:creator>
<dc:creator>Alencar, G. F.</dc:creator>
<dc:creator>Pulliam, T.</dc:creator>
<dc:creator>McClure, E.</dc:creator>
<dc:creator>Singh, N.</dc:creator>
<dc:creator>Urselli, F.</dc:creator>
<dc:creator>Poljakov, K.</dc:creator>
<dc:creator>Smythe, K. S.</dc:creator>
<dc:creator>Kulikauskas, R. M.</dc:creator>
<dc:creator>Robinson, K. L.</dc:creator>
<dc:creator>Moshiri, A. S.</dc:creator>
<dc:creator>Yeung, C. C.</dc:creator>
<dc:creator>Lin, M.</dc:creator>
<dc:creator>Shimp, K. R.</dc:creator>
<dc:creator>Schwartz, A.</dc:creator>
<dc:creator>Macy, A. M.</dc:creator>
<dc:creator>Tooley, M. R.</dc:creator>
<dc:creator>Baker, M. L.</dc:creator>
<dc:creator>Carter, J. J.</dc:creator>
<dc:creator>Hopwo, K.</dc:creator>
<dc:creator>Singhi, N.</dc:creator>
<dc:creator>Bakhtiari, J.</dc:creator>
<dc:creator>Ruterbusch, M.</dc:creator>
<dc:creator>Shasha, C. G.</dc:creator>
<dc:creator>Iuliano, M.</dc:creator>
<dc:creator>Mullen, L. J.</dc:creator>
<dc:creator>Debuysscher, B. L.</dc:creator>
<dc:creator>Veatch, J. R.</dc:creator>
<dc:creator>Koelle, D. M.</dc:creator>
<dc:creator>Galloway, D. A.</dc:creator>
<dc:creator>Nghiem, P.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:date>2025-07-24</dc:date>
<dc:identifier>doi:10.1101/2025.07.22.666002</dc:identifier>
<dc:title><![CDATA[B cells specific for polyomavirus-derived oncoprotein are predictive of Merkel cell carcinoma progression]]></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/10.1101/2025.07.28.667202v1?rss=1">
<title>
<![CDATA[
Q4ddPCR (May the Fourth Be Precise): A Flexible, 4-Target Assay for High-Resolution HIV Reservoir Profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.28.667202v1?rss=1</link>
<description><![CDATA[
Precise and scalable quantification of the genetically intact HIV reservoir is critical for advancing curative strategies. However, current HIV reservoir assays such as the intact proviral DNA assay (IPDA) are limited by quantification failures or misclassification of defective proviral genomes due to HIV sequence heterogeneity. Q4ddPCR is a modular, droplet digital PCR assay that simultaneously targets four conserved regions in the HIV genome to improve specificity, reduce quantification gaps, and provide multi-layered readouts. We benchmarked Q4ddPCR against 3,650 near full-length proviral sequences from 13 virally suppressed people with HIV (PWH) generated by Q4PCR using the same primer/probe sets. Q4ddPCR enabled intact reservoir quantification in 95% of samples from three independent cohorts and closely matched sequence-confirmed Q4PCR reservoir measurements. In addition, multi-probe readouts revealed clonal intact reservoir dynamics that are not detectable by IPDA. In longitudinal samples from 42 participants over the first 4.5 years on antiretroviral therapy (ART), Q4ddPCR reported lower proviral frequencies and a steeper decline in intact proviral DNA compared to IPDA. Collectively, our findings confirm key predictions from mathematical modeling, demonstrating that multi-target assays provide greater specificity and more accurately capture the dynamics of the intact HIV reservoir.
]]></description>
<dc:creator>Scheck, R.</dc:creator>
<dc:creator>Melzer, M.</dc:creator>
<dc:creator>Gladkov, G.</dc:creator>
<dc:creator>Ward, A. R.</dc:creator>
<dc:creator>Reeves, D. B.</dc:creator>
<dc:creator>Perkins, N.</dc:creator>
<dc:creator>Huynh, T. T.</dc:creator>
<dc:creator>McMahon, D. K.</dc:creator>
<dc:creator>Bosch, R. J.</dc:creator>
<dc:creator>Macatangay, B. J.</dc:creator>
<dc:creator>Cyktor, J. C.</dc:creator>
<dc:creator>Eron, J. J.</dc:creator>
<dc:creator>Mellors, J. W.</dc:creator>
<dc:creator>Gandhi, R. T.</dc:creator>
<dc:creator>Buchauer, L.</dc:creator>
<dc:creator>Jones, R. B.</dc:creator>
<dc:creator>Gaebler, C.</dc:creator>
<dc:date>2025-07-31</dc:date>
<dc:identifier>doi:10.1101/2025.07.28.667202</dc:identifier>
<dc:title><![CDATA[Q4ddPCR (May the Fourth Be Precise): A Flexible, 4-Target Assay for High-Resolution HIV Reservoir Profiling]]></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/10.1101/2025.07.29.667532v1?rss=1">
<title>
<![CDATA[
Cellular heterogeneity and therapeutic response profiling of human IDH+ glioma stem cell cultures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.29.667532v1?rss=1</link>
<description><![CDATA[
Summary/AbstractGlioblastoma stem cell (GSC) cultures are initiated from glioblastoma (GBM) surgical resection tissue. They can capture and propagate key GBM primary tumor molecular and cellular features. We have deeply characterized four isocitrate dehydrogenase (IDH)-expressing (or IDH+) GSC cultures from unrelated adults to serve as cellular models for the majority of adult primary GBM. We demonstrate that GSC cultures can be continuously propagated in defined, serum-free media and 5% oxygen without requiring specialized growth substrates; have well-defined genomic and mtDNA variants and gene/protein expression profiles; and highly reproducible dose-survival curves when treated with the GBM standard-of-care therapies of ionizing radiation (IR) and temozolomide (TMZ). We also illustrate how expressed lentiviral barcodes, mtDNA variants and single cell gene expression profiling can be used to define and track cellular heterogeneity over 40 days after IR treatment. These well-characterized IDH+ GSC cultures can support many high throughput in vitro assay formats, including xenograft, organoid and other GBM disease modeling protocols. They should prove a useful resource to better understand GBM biology, and to identify new and more effective GBM therapies and treatment regimens.

HighlightsO_LIGlioblastoma (GBM)-derived IDH-expressing Glioma Stem Cell (GSC) cultures can capture and propagate GBM genomic variants, gene and protein expression programs.
C_LIO_LIMitochondrial DNA (mtDNA) variants identified by high accuracy Duplex DNA sequencing are abundant and sub-clonally organized in GSC cultures.
C_LIO_LIGSC cultures have highly reproducible dose-survival curves for ionizing radiation and temozolomide, the GBM standard-of-care therapies.
C_LIO_LIGSC culture cellular heterogeneity can be captured, characterized and tracked by using expressed lentiviral barcodes, mtDNA variants and scRNA sequencing.
C_LI

eTOC blurbIDH-expressing glioma stem cell cultures (GSCs) are experimentally tractable and versatile glioblastoma (GBM) cellular disease models. Deeply characterized GSC cultures can enable new work to understand GBM biology, and help identify new and more effective GBM therapies and treatment regimens.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/667532v1_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@82aceeorg.highwire.dtl.DTLVardef@1277c18org.highwire.dtl.DTLVardef@144188eorg.highwire.dtl.DTLVardef@1a84bb3_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Chambwe, N.</dc:creator>
<dc:creator>Kennedy, S. R.</dc:creator>
<dc:creator>Kohrn, B. F.</dc:creator>
<dc:creator>Lazarchuk, P.</dc:creator>
<dc:creator>Leutert, M.</dc:creator>
<dc:creator>Qin, G.</dc:creator>
<dc:creator>Tercan, B.</dc:creator>
<dc:creator>Sanchez-Contreras, M.</dc:creator>
<dc:creator>Tang, W.</dc:creator>
<dc:creator>Graber, J.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:creator>Villen, J.</dc:creator>
<dc:creator>Shmulevich, I.</dc:creator>
<dc:creator>Monnat, R. J.</dc:creator>
<dc:date>2025-08-01</dc:date>
<dc:identifier>doi:10.1101/2025.07.29.667532</dc:identifier>
<dc:title><![CDATA[Cellular heterogeneity and therapeutic response profiling of human IDH+ glioma stem cell cultures]]></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/10.1101/2025.07.30.667762v1?rss=1">
<title>
<![CDATA[
Stabilization of H5 highly pathogenic avian influenza hemagglutinin improves vaccine-elicited neutralizing antibody responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.30.667762v1?rss=1</link>
<description><![CDATA[
Transmission of highly pathogenic avian influenza from H5 clade 2.3.4.4b has expanded in recent years to infect large populations of birds and mammals, heightening the risk of a human pandemic. Influenza viruses adapted to transmission in birds and some other animals tend to have a less stable hemagglutinin (HA) than seasonal influenza viruses, enabling membrane fusion at comparatively high pH levels. Here, we combine five mutations within H5 HA that dramatically increase its melting temperature and promote stable closure of the HA trimer. Structural analysis by cryo-electron microscopy revealed that the stabilizing mutations create several new hydrophobic interactions, while maintaining local HA structure. We found that vaccinating mice with stabilized H5 HA immunogens resulted in higher hemagglutination inhibition and neutralization titers than non-stabilized comparators. Epitope mapping of vaccine-elicited polyclonal antibody responses using negative stain electron microscopy and deep mutational scanning showed that site E on the side of the HA receptor binding domain was immunodominant across all groups; however, the stabilized immunogens shifted responses toward the receptor binding site (RBS), eliciting a higher proportion of neutralizing antibodies. These findings highlight that H5 HA-stabilizing mutations enhance the quality of antibody responses across different vaccine formats, underscoring their potential to improve pandemic preparedness vaccines targeting viruses from this widely circulating clade.

One Sentence SummaryVaccinating with stabilized H5 HA increases the proportion of humoral responses against neutralizing RBS epitopes.
]]></description>
<dc:creator>Dosey, A.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Gillespie, R. A.</dc:creator>
<dc:creator>Leaf, E. M.</dc:creator>
<dc:creator>Vukovich, M. J.</dc:creator>
<dc:creator>McGowan, J.</dc:creator>
<dc:creator>Grey, E.</dc:creator>
<dc:creator>Muramatsu, H.</dc:creator>
<dc:creator>Jun, R. H.</dc:creator>
<dc:creator>Pardi, N.</dc:creator>
<dc:creator>Kanekiyo, M.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:date>2025-08-02</dc:date>
<dc:identifier>doi:10.1101/2025.07.30.667762</dc:identifier>
<dc:title><![CDATA[Stabilization of H5 highly pathogenic avian influenza hemagglutinin improves vaccine-elicited neutralizing antibody responses]]></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/10.1101/2025.08.07.669162v1?rss=1">
<title>
<![CDATA[
A bone marrow stromal secretome screen identifies semaphorin 3A as a regulator of hematopoiesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.07.669162v1?rss=1</link>
<description><![CDATA[
Bone marrow mesenchymal stromal cells (MSCs) are a major source of secreted factors that control hematopoietic stem and progenitor cell (HSPC) function. We previously reported the generation of revitalized MSCs (rMSCs), which more effectively support HSPCs in culture. In a secretome screen using rMSCs, we identified semaphorin 3A (SEM3A) as a secreted factor upregulated as part of a pro-inflammatory signature that may contribute to HSPC expansion by rMSCs. We show that recombinant SEM3A acts directly on HSPCs to inhibit their cycling ex vivo. Analysis of a SEM3A loss of function mutation in vivo revealed hematopoietic progenitor expansion and accelerated recovery after myeloablation, consistent with a role for SEM3A in regulating HSPCs at steady state and during hematopoietic stress. This work highlights proteomic screening using rMSCs as a method to identify novel secreted niche factors and uncovers a novel role for SEM3A in controlling HSPC proliferation in stress hematopoiesis.

SummaryBorger et al. characterize the secretome of revitalized bone marrow stromal cells and identify a novel role of the protein semaphorin 3A in regulating hematopoietic stem and progenitor cell proliferation in steady state and stress conditions.
]]></description>
<dc:creator>Borger, D. K.</dc:creator>
<dc:creator>Mitchell, S. P.</dc:creator>
<dc:creator>Nakahara, F.</dc:creator>
<dc:creator>Colombo, D. F.</dc:creator>
<dc:creator>Torres, L. S.</dc:creator>
<dc:creator>Garforth, S. J.</dc:creator>
<dc:creator>Sidoli, S.</dc:creator>
<dc:creator>Krijgsveld, J.</dc:creator>
<dc:creator>Silberstein, L.</dc:creator>
<dc:creator>Frenette, P. S.</dc:creator>
<dc:creator>Gritsman, K.</dc:creator>
<dc:date>2025-08-07</dc:date>
<dc:identifier>doi:10.1101/2025.08.07.669162</dc:identifier>
<dc:title><![CDATA[A bone marrow stromal secretome screen identifies semaphorin 3A as a regulator of hematopoiesis]]></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/10.1101/2025.08.08.668693v1?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/10.1101/2025.08.12.669944v1?rss=1">
<title>
<![CDATA[
RHOV is a Detachment-Responsive Rho GTPase Necessary for Ovarian Cancer Peritoneal Metastasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.12.669944v1?rss=1</link>
<description><![CDATA[
All ovarian cancer subtypes spread via transcoelomic metastasis, where cells disseminate into the peritoneal fluid, resist anoikis, and form multicellular aggregates that invade the peritoneum. This represents the main driver of morbidity and mortality for peritoneal cancer patients. Mechanisms necessary for cancer cells to survive matrix detachment and initiate transcoelomic metastasis remain poorly defined. To address this, we identified a conserved detachment-sensitive gene signature activated shortly after matrix-detachment across multiple ascites-derived cancer cell lines. RHOV, an atypical, constitutively active and understudied member of the Rho GTPase family, emerged as a top upregulated transcript, which was confirmed in patient ascites-derived tumor cells. Functionally, loss of RHOV impairs anoikis resistance, multicellular aggregate integrity, migration and invasion, and completely abolishes transcoelomic tumor progression in vivo. RHOV enhances c-Jun signaling and cytoskeletal remodeling, which is dependent on both RHOV GTP-binding and membrane localization. These findings define RHOV as a novel detachment-sensitive Rho GTPase and establish RHOV as a critical regulator of peritoneal metastasis for the first time.
]]></description>
<dc:creator>Elhaw, A. T.</dc:creator>
<dc:creator>Tang, P. W.</dc:creator>
<dc:creator>Cheng, Y.-Y.</dc:creator>
<dc:creator>Kamlapurkar, S.</dc:creator>
<dc:creator>Javed, Z.</dc:creator>
<dc:creator>Al-Saad, S.</dc:creator>
<dc:creator>White, S. R.</dc:creator>
<dc:creator>Abdelnaby, A. E.</dc:creator>
<dc:creator>Khan, H.</dc:creator>
<dc:creator>Choi, A. S.</dc:creator>
<dc:creator>Cole, A. R.</dc:creator>
<dc:creator>Kim, Y.-S.</dc:creator>
<dc:creator>Atiya, H. I.</dc:creator>
<dc:creator>Trebak, M.</dc:creator>
<dc:creator>Zervantonakis, I. K.</dc:creator>
<dc:creator>Buckanovich, R. J.</dc:creator>
<dc:creator>Aird, K. M.</dc:creator>
<dc:creator>Coffman, L. G.</dc:creator>
<dc:creator>Mythreye, K.</dc:creator>
<dc:creator>Hempel, N.</dc:creator>
<dc:date>2025-08-13</dc:date>
<dc:identifier>doi:10.1101/2025.08.12.669944</dc:identifier>
<dc:title><![CDATA[RHOV is a Detachment-Responsive Rho GTPase Necessary for Ovarian Cancer Peritoneal Metastasis]]></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/10.1101/2025.08.12.669682v1?rss=1">
<title>
<![CDATA[
Pro-inflammatory role of granzyme K producing bystander CD8+ T cells in acute myeloid leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.12.669682v1?rss=1</link>
<description><![CDATA[
Acute myeloid leukemia (AML) is a heterogeneous group of blood malignancies with a 5-year survival rate below 30%, highlighting the urgent need for more effective therapeutic strategies. T cell-based immunotherapies have demonstrated remarkable success in solid tumors, yet the role of CD8+ T cells in AML remains unclear. In this study, we analyzed the composition, antigenic specificity, and function of CD8+ T cells in paired blood and bone marrow samples from AML patients. While we did not identify exhausted CD8+ T cells as seen in solid tumors, we observed a distinct population of functional CD69+ CD8+ T cells specifically enriched in the bone marrow. These cells primarily recognized non-tumor antigens, including epitopes derived from Epstein-Barr virus (EBV) and cytomegalovirus (CMV). Notably, this bystander CD8+ T cell population showed high expression of Granzyme K, a cytokine found in the bone marrow of AML patients. Granzyme K did not induce leukemic cell death but instead promoted the secretion of IL-8, a pro-inflammatory cytokine known to play a detrimental role in AML pathology. Rather than mounting an anti-tumor response, these CD8+ T cells contribute to a pro-inflammatory environment that may exacerbate AML progression and severity. These findings provide a rationale for exploring therapeutic strategies aimed at inhibiting pro-inflammatory CD8+ T cells and targeting Granzyme K activity in association with actual therapies.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=137 SRC="FIGDIR/small/669682v1_ufig1.gif" ALT="Figure 1">
View larger version (32K):
org.highwire.dtl.DTLVardef@1fc9798org.highwire.dtl.DTLVardef@4491b1org.highwire.dtl.DTLVardef@1b8feadorg.highwire.dtl.DTLVardef@58515d_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Aziez, L.</dc:creator>
<dc:creator>Deredec, N.</dc:creator>
<dc:creator>Boussaid, I.</dc:creator>
<dc:creator>Shasha, C. G.</dc:creator>
<dc:creator>Vazquez, R.</dc:creator>
<dc:creator>Friedrich, C.</dc:creator>
<dc:creator>Alik, A.</dc:creator>
<dc:creator>Manasse, K.</dc:creator>
<dc:creator>Fremont-Debaene, Z.</dc:creator>
<dc:creator>Barthelemy, A.</dc:creator>
<dc:creator>Catelain, C.</dc:creator>
<dc:creator>Rameau, P.</dc:creator>
<dc:creator>Vignon, M.</dc:creator>
<dc:creator>Decroocq, J.</dc:creator>
<dc:creator>Kosmider, O.</dc:creator>
<dc:creator>Selimoglu-Buet, D.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Solary, E.</dc:creator>
<dc:creator>Delhommeau, F.</dc:creator>
<dc:creator>Herault, O.</dc:creator>
<dc:creator>TARTOUR, E.</dc:creator>
<dc:creator>Birsen, R.</dc:creator>
<dc:creator>Bouscary, D.</dc:creator>
<dc:creator>Fontenay, M.</dc:creator>
<dc:creator>Chapuis, N.</dc:creator>
<dc:creator>Simoni, Y.</dc:creator>
<dc:date>2025-08-15</dc:date>
<dc:identifier>doi:10.1101/2025.08.12.669682</dc:identifier>
<dc:title><![CDATA[Pro-inflammatory role of granzyme K producing bystander CD8+ T cells in acute myeloid leukemia]]></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/10.1101/2025.08.14.670174v1?rss=1">
<title>
<![CDATA[
MDS-associated SF3B1 mutations promote aberrant fate choice of hematopoietic stem cell via mis-splicing of mediator kinase module component CDK8 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.14.670174v1?rss=1</link>
<description><![CDATA[
Mutations in RNA splicing factor SF3B1 are among the most common in MDS and are strongly associated with MDS with ring sideroblasts (MDS-RS). While aberrant splicing of terminal erythroid regulators has been implicated in MDS pathogenesis, the impact of SF3B1 mutations on early hematopoietic progenitor function remains unclear. Here, we identify CDK8, a key kinase of the mediator complex involved in transcriptional regulation, as a recurrent mis-spliced target in SF3B1-mutant MDS. Mutant SF3B1 induces cryptic 3' splice site selection in CDK8, leading to loss of CDK8 mRNA and protein. Using primary human HSPCs, our study identifies CDK8 as an important regulator of HSPC homeostasis and cell fate determination. CDK8 depletion results in expansion of HSPCs and shifts differentiation toward the erythroid and myeloid lineages, mirroring phenotypes observed in SF3B1-mutant MDS. Lastly, functional restoration of CDK8 rescues early erythroid phenotypes in SF3B1-mutant cells. These findings implicate CDK8 mis-splicing as a mechanistic driver of altered progenitor fate and dysplasia in SF3B1-mutant MDS, linking aberrant splicing to transcriptional dysregulation and hematopoietic lineage commitment.
]]></description>
<dc:creator>Bonner, E. A.</dc:creator>
<dc:creator>Hsueh, T.-Y.</dc:creator>
<dc:creator>Song, A.</dc:creator>
<dc:creator>Arriaga-Gomez, E. A.</dc:creator>
<dc:creator>Venkataraman, R.</dc:creator>
<dc:creator>Sinha, S.</dc:creator>
<dc:creator>Nguyen, E.</dc:creator>
<dc:creator>Ferrell, P. B.</dc:creator>
<dc:creator>Welner, R. S.</dc:creator>
<dc:creator>Lu, R.</dc:creator>
<dc:creator>Stirewalt, D. L.</dc:creator>
<dc:creator>Doulatov, S.</dc:creator>
<dc:creator>Lee, S. C.</dc:creator>
<dc:date>2025-08-19</dc:date>
<dc:identifier>doi:10.1101/2025.08.14.670174</dc:identifier>
<dc:title><![CDATA[MDS-associated SF3B1 mutations promote aberrant fate choice of hematopoietic stem cell via mis-splicing of mediator kinase module component CDK8]]></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/10.1101/2025.08.18.671001v1?rss=1">
<title>
<![CDATA[
Spike mutations that affect the function and antigenicity of recent KP.3.1.1-like SARS-CoV-2 variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.18.671001v1?rss=1</link>
<description><![CDATA[
SARS-CoV-2 is under strong evolutionary selection to acquire mutations in its spike protein that reduce neutralization by human polyclonal antibodies. Here we use pseudovirus-based deep mutational scanning to measure how mutations to the spike from the recent KP.3.1.1 SARS-CoV-2 strain affect cell entry, binding to ACE2 receptor, RBD up/down motion, and neutralization by human sera and clinically relevant antibodies. The spike mutations that most affect serum antibody neutralization sometimes differ between sera collected before versus after recent vaccination or infection, indicating these exposures shift the neutralization immunodominance hierarchy. The sites where mutations cause the greatest reduction in neutralization by post-vaccination or infection sera include receptor-binding domain (RBD) sites 475, 478 and 487, all of which have mutated in recent SARS-CoV-2 variants. Multiple mutations outside the RBD affect sera neutralization as strongly as any RBD mutations by modulating RBD up/down movement. Some sites that affect RBD up/down movement have mutated in recent SARS-CoV-2 variants. Finally, we measure how spike mutations affect neutralization by three clinically relevant SARS-CoV-2 antibodies: VYD222, BD55-1205, and SA55. Overall, these results illuminate the current constraints and pressures shaping SARS-CoV-2 evolution, and can help with efforts to forecast possible future antigenic changes that may impact vaccines or clinical antibodies.

ImportanceThis study measures how mutations to the spike of a SARS-CoV-2 variant that circulated in early 2025 affect its function and recognition by both the polyclonal antibodies produced by the human immune system and monoclonal antibodies used as prophylactics. These measurements are made with a pseudovirus system that enables safe study of viral protein mutations using virions that can only infect cells once. The study identifies mutations that decrease recognition by current human antibody immunity; many of these mutations are increasingly being observed in new viral variants. It also shows the importance of mutations that move the spikes receptor binding domain up or down. Overall, these results are useful for forecasting viral evolution and assessing which newly emerging variants have reduced recognition by immunity and antibody prophylactics.
]]></description>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Harari, S.</dc:creator>
<dc:creator>Larsen, B. B.</dc:creator>
<dc:creator>Kampman, L.</dc:creator>
<dc:creator>Harteloo, A.</dc:creator>
<dc:creator>Elias-Warren, A.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2025-08-19</dc:date>
<dc:identifier>doi:10.1101/2025.08.18.671001</dc:identifier>
<dc:title><![CDATA[Spike mutations that affect the function and antigenicity of recent KP.3.1.1-like SARS-CoV-2 variants]]></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/10.1101/2025.08.21.671404v1?rss=1">
<title>
<![CDATA[
An emergent disease-associated motor neuron state precedes cell death in a mouse model of ALS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.21.671404v1?rss=1</link>
<description><![CDATA[
To uncover molecular determinants of motor neuron degeneration and selective vulnerability in amyotrophic lateral sclerosis (ALS), we generated longitudinal single-nucleus transcriptomes and chromatin accessibility profiles of spinal motor neurons from the SOD1-G93A ALS mouse model. Vulnerable alpha motor neurons showed thousands of molecular changes, marking a transition into a novel cell state we named  disease-associated motor neurons (DAMNs). We identified transcription factor regulatory networks that govern how healthy cells transition into DAMNs as well as those linked to vulnerable and resistant motor neuron subtypes. Using spatial transcriptomics, we found reactive glia located near motor neurons early in disease, suggesting early signaling events between motor neurons and glia. Finally, we found that the human orthologs of genomic regions with differential accessibility in SOD1-G93A alpha motor neurons are enriched for single nucleotide polymorphisms associated with human ALS, providing evidence that the genetic underpinnings of motor neuron vulnerability are conserved.
]]></description>
<dc:creator>Gautier, O.</dc:creator>
<dc:creator>Blum, J. A.</dc:creator>
<dc:creator>Nguyen, T. P.</dc:creator>
<dc:creator>Klemm, S.</dc:creator>
<dc:creator>Yamakawa, M.</dc:creator>
<dc:creator>Sinnott-Armstrong, N.</dc:creator>
<dc:creator>Zeng, Y.</dc:creator>
<dc:creator>Davis, C.-H. O.</dc:creator>
<dc:creator>Bombosch, J.</dc:creator>
<dc:creator>Nakayama, L.</dc:creator>
<dc:creator>Guttenplan, K. A.</dc:creator>
<dc:creator>Chen, D.</dc:creator>
<dc:creator>Kathira, A.</dc:creator>
<dc:creator>Zhao, L. Y.</dc:creator>
<dc:creator>Rexach, J. E.</dc:creator>
<dc:creator>Greenleaf, W. E.</dc:creator>
<dc:creator>Gitler, A. D.</dc:creator>
<dc:date>2025-08-25</dc:date>
<dc:identifier>doi:10.1101/2025.08.21.671404</dc:identifier>
<dc:title><![CDATA[An emergent disease-associated motor neuron state precedes cell death in a mouse model of ALS]]></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/10.1101/2025.08.26.671106v1?rss=1">
<title>
<![CDATA[
An Efficient Lasso Framework for Admixture-Aware Polygenic Scores 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.26.671106v1?rss=1</link>
<description><![CDATA[
Polygenic scores (PGS) have promising clinical applications for risk stratification, disease screening, and personalized medicine. However, most PGS are trained on predominantly European ancestry cohorts and have limited portability to external populations. While cross-population PGS methods have demonstrated greater generalizability than single-ancestry PGS, they fail to properly account for individuals with recent admixture between continental ancestry groups. GAUDI is a recently proposed PGS method which overcomes this gap by leveraging local ancestry to estimate ancestry-specific effects, penalizing but allowing ancestry-differential effects. However, the modified fused LASSO approach used by GAUDI is computationally expensive and does not readily accommodate more than two-way admixture. To address these limitations, we introduce HAUDI, an efficient LASSO framework for admixed PGS construction. HAUDI re-parameterizes the GAUDI model as a standard LASSO problem, allowing for extension to multi-way admixture settings and far superior computational speed than GAUDI. In extensive simulations, HAUDI compares favorably to GAUDI while dramatically reducing computation time. In real data applications, HAUDI uniformly out-performs GAUDI across 18 clinical phenotypes, including total triglycerides (TG), C-reactive protein (CRP), and mean corpuscular hemoglobin concentration (MCHC), and shows substantial benefits over an ancestry-agnostic PGS for white blood cell count (WBC) and chronic kidney disease (CKD).
]]></description>
<dc:creator>Ockerman, F. P.</dc:creator>
<dc:creator>Chen, B. D.</dc:creator>
<dc:creator>Sun, Q.</dc:creator>
<dc:creator>Kharitonova, E. V.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Zhou, L. Y.</dc:creator>
<dc:creator>Loos, R. J.</dc:creator>
<dc:creator>Kooperberg, C.</dc:creator>
<dc:creator>Peters, U.</dc:creator>
<dc:creator>Haessler, J.</dc:creator>
<dc:creator>Reiner, A. P.</dc:creator>
<dc:creator>Jung, S. Y.</dc:creator>
<dc:creator>Manson, J. E.</dc:creator>
<dc:creator>Nassir, R.</dc:creator>
<dc:creator>North, K. E.</dc:creator>
<dc:creator>Buyske, S.</dc:creator>
<dc:creator>Haiman, C. A.</dc:creator>
<dc:creator>Conti, D. V.</dc:creator>
<dc:creator>Wilkens, L. R.</dc:creator>
<dc:creator>Lange, E. M.</dc:creator>
<dc:creator>Cox, N. J.</dc:creator>
<dc:creator>Cao, H.</dc:creator>
<dc:creator>Raffield, L. M.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Tao, R.</dc:creator>
<dc:date>2025-08-27</dc:date>
<dc:identifier>doi:10.1101/2025.08.26.671106</dc:identifier>
<dc:title><![CDATA[An Efficient Lasso Framework for Admixture-Aware Polygenic Scores]]></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/10.1101/2025.08.28.672949v1?rss=1">
<title>
<![CDATA[
Determinants of Natural Killer Cell-Mediated Antibody Dependent Cellular Cytotoxicity in SARS-CoV-2 Antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.28.672949v1?rss=1</link>
<description><![CDATA[
A growing body of evidence underscores the role of antibody-dependent cellular cytotoxicity (ADCC) in antiviral immunity. Yet, the mechanisms underlying the ability of certain antibodies (Abs) to mediate potent ADCC activity remain poorly characterized - in particular the contribution of features within the antigen binding Fab region remain largely unexplored. In this study, we leveraged a collection of 142 SARS-CoV2 monoclonal Abs to systematically dissect the determinants of ADCC activity. We analyzed their epitope domain target, binding characteristics, neutralization potency, somatic hypermutation (SHM) and CDR3 length to determine the contribution of these features to ADCC activity. We found that ADCC activity is primarily shaped by epitope target-particularly targeting of the S2 domain of the Spike glycoprotein. ADCC potency was not associated with the degree of SHM or neutralization. Notably, ADCC activity was not correlated with binding affinity and moderate binding to antigen was sufficient for ADCC activity. By integrating these analyses, we provide a comprehensive framework for understanding the molecular and functional determinants of ADCC. Together, these findings offer novel insights into the mechanisms that underpin ADCC functions, with implications for vaccine design and therapeutic Abs development.

Author SummaryWhile antibodies are best known for stopping viruses from entering cells, they also activate other arms of the immune system. One important function is their ability to signal immune cells to destroy virus-infected targets, a process called antibody-dependent cellular cytotoxicity (ADCC). In this study, we examined 142 antibodies directed against the spike protein of the virus that causes COVID-19. We compared their binding patterns, neutralizing ability, level of maturation, and other features to determine what drives their capacity to trigger ADCC. We found that the region of the spike protein targeted by the antibody was the main factor shaping this activity, with antibodies that recognized the S2 region being particularly effective. In contrast, the amount of antibody maturation, the strength of binding, or the ability to neutralize the virus did not predict immune cell activation. Our findings show that antibodies can contribute to antiviral defense through distinct mechanisms and provide new insights for guiding vaccine and antibody therapy design.
]]></description>
<dc:creator>Depierreux, D. M.</dc:creator>
<dc:creator>Ruiz, F. M.</dc:creator>
<dc:creator>Lilly, M.</dc:creator>
<dc:creator>Guenthoer, J.</dc:creator>
<dc:creator>Chohan, V.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2025-08-29</dc:date>
<dc:identifier>doi:10.1101/2025.08.28.672949</dc:identifier>
<dc:title><![CDATA[Determinants of Natural Killer Cell-Mediated Antibody Dependent Cellular Cytotoxicity in SARS-CoV-2 Antibodies]]></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/10.1101/2025.08.29.673185v1?rss=1">
<title>
<![CDATA[
Distinct modes of evolution drive HIV escape from two broadly neutralizing antibodies. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.29.673185v1?rss=1</link>
<description><![CDATA[
Broadly neutralizing antibodies (bNAbs) show promise for HIV treatment and prevention, but are vulnerable to resistance evolution. Comprehensively understanding in vivo viral escape from individual bNAbs is necessary to design bNAb combinations that will provide durable responses. We characterize viral escape from two such bNAbs, 10-1074 and 3BNC117, using deep, longitudinal sequencing of full length HIV envelope (env) genes from study participants treated with bNAb monotherapy. Improved sequencing depth and computational evolutionary analyses permit us to identify in vivo routes and parallelism underlying HIV escape from each bNAb, providing new insights into this evolutionary process. We find that 10-1074 escape is restricted to a small number of previously documented pathways seen across participants, but these escape mutations 1) emerge via extensively recurrent mutation, 2) are not equally preferred, and 3) can pre-exist at low frequency in intra-host viral populations before therapy, although their detection does not predict rebound timing. In contrast, 3BNC117 escape follows background-specific patterns in which specific escape mutations present in one intra-host population rarely emerge or spread in other populations, except among highly related viruses. Despite this, 3BNC117 escape mutations often still emerge recurrently within their host. Our findings map longitudinal in vivo antibody escape across 20 diverse clade B HIV intra-host populations and reveal clinically relevant resistance dynamics that highlight how combination bNAb therapies will need to contend with extensively recurring escape mutations and dependence on genetic background.

Significance StatementUsing recently developed techniques that capture viral genetic diversity and associations between mutations at depth, we deeply sequenced HIV from two clinical trials of broadly neutralizing antibody (bNAb) monotherapies, 3BNC117 and 10-1074. We computationally characterized HIV populations longitudinally with unprecedented resolution as they escaped these therapies in people living with HIV. Intra-host tracking of individual HIV genetic backgrounds reveals extensively recurrent mutations driving escape and suggests that HIV escape routes from certain bNAbs can depend sensitively on the genetic background of the virus. Our findings highlight the difficulties in evaluating pre-treatment resistance, provide an analysis blueprint for future trials, and inform the design of emerging combination antibody therapies to maximize the likelihood of durable efficacy.
]]></description>
<dc:creator>Romero, E. V.</dc:creator>
<dc:creator>Clyde, A. E.</dc:creator>
<dc:creator>Giorgi, E. E.</dc:creator>
<dc:creator>Westfall, D. H.</dc:creator>
<dc:creator>Azam, W.</dc:creator>
<dc:creator>Taylor, M. L.</dc:creator>
<dc:creator>Caskey, M.</dc:creator>
<dc:creator>Feder, A. F.</dc:creator>
<dc:creator>Cohn, L. B.</dc:creator>
<dc:date>2025-08-30</dc:date>
<dc:identifier>doi:10.1101/2025.08.29.673185</dc:identifier>
<dc:title><![CDATA[Distinct modes of evolution drive HIV escape from two broadly neutralizing antibodies.]]></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/10.1101/2025.09.03.674087v1?rss=1">
<title>
<![CDATA[
Rapid protamine evolution suppresses meiotic drive in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.03.674087v1?rss=1</link>
<description><![CDATA[
Many animal species replace histones with protamines during spermatogenesis. Despite their importance for sperm function, protamines rapidly evolve in many species; the biological causes behind their rapid evolution remain unknown. Here, using in vivo gene replacement, we investigated the causes and consequences underlying the rapid evolution of protamine Mst77F, which is essential for male fertility in D. melanogaster. Mst77F ortholog replacements led to defects in DNA compaction of X-chromosome-bearing sperm compared to Y-chromosome-bearing sperm during spermatogenesis, resulting in fewer X-bearing mature sperm and male-biased progeny. Unlike D. melanogaster, Mst77F is not essential for male fertility in D. yakuba but is still required to suppress sex-ratio distortion. Our results suggest that relentless pressure to suppress sex chromosomal meiotic drive drives the rapid evolution of protamines.

One-sentence summaryA rapidly evolving essential protamine suppresses sex-chromosome meiotic drive in Drosophila
]]></description>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>de la Cruz, A. F.</dc:creator>
<dc:creator>Natividad, I. M.</dc:creator>
<dc:creator>Noyola, A.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2025-09-05</dc:date>
<dc:identifier>doi:10.1101/2025.09.03.674087</dc:identifier>
<dc:title><![CDATA[Rapid protamine evolution suppresses meiotic drive in Drosophila]]></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/10.1101/2025.09.02.673849v1?rss=1">
<title>
<![CDATA[
Transcriptional Rewiring of BET Inhibitor Treated Ewing Sarcoma Cells Augments their Dependency on Focal Adhesion Kinase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.02.673849v1?rss=1</link>
<description><![CDATA[
Epigenetic dysregulation is a hallmark of cancer. Small molecule inhibitors such as bromodomain and extraterminal (BET) protein inhibitors developed to target epigenetic dependencies have demonstrated significant promise in preclinical models. However, clinical success with epigenetic drugs as single agents has been limited by emergence of tumor cell tolerance and escape, which often occurs due to transcriptional rewiring. Ewing sarcoma (EwS), a bone and soft tissue tumor driven by the EWS::FLI1 fusion oncoprotein, is characterized by profound oncogene-dependent enhancer reprogramming. Thus, epigenetic modifying therapies are of high therapeutic interest. In this study, we sought to elucidate how EwS cells escape BET inhibition to identify biologically informed drug combinations that could be advanced to clinical trials. As expected, EwS cells and xenografts initially responded to BMS-986158, a pharmaceutical grade BET inhibitor, but proliferation was rapidly restored. A kinase inhibitor screen showed that BMS-986158 drug tolerant persister (DTP) cells were sensitive to inhibitors of Focal Adhesion Kinase (FAK), a critical signaling node downstream of extracellular matrix (ECM) engagement. RNA sequencing revealed that DTP cells had been transcriptionally rewired and that mesenchymal signature and ECM remodeling genes were specifically upregulated. Combining BMS-986158 with the FAK inhibitor Defactinib had synergistic effects, reducing EwS cell proliferation, survival, and invasion in vitro, and significantly inhibited tumor outgrowth in vivo. Our studies identify BET and FAK inhibition as a rational combination therapy worthy of further investigation for EwS, and demonstrate that defining emergent mechanisms of epigenetic drug tolerance can identify new vulnerabilities that can be therapeutically targeted.
]]></description>
<dc:creator>Ganapathi, S. S.</dc:creator>
<dc:creator>Wrenn, E. D.</dc:creator>
<dc:creator>Garcia, N. M.</dc:creator>
<dc:creator>Katiyar, N.</dc:creator>
<dc:creator>Walter, S. I.</dc:creator>
<dc:creator>Miyaki, A.</dc:creator>
<dc:creator>Valenti, J.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:creator>Lawlor, E. R.</dc:creator>
<dc:date>2025-09-06</dc:date>
<dc:identifier>doi:10.1101/2025.09.02.673849</dc:identifier>
<dc:title><![CDATA[Transcriptional Rewiring of BET Inhibitor Treated Ewing Sarcoma Cells Augments their Dependency on Focal Adhesion Kinase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.03.674106v1?rss=1">
<title>
<![CDATA[
MondoA mediates transcriptional coordination between the MYC network and the integrated stress response in pancreatic ductal adenocarcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.03.674106v1?rss=1</link>
<description><![CDATA[
MYC amplification contributes to poor survival and outcome in pancreatic ductal adenocarcinoma (PDAC). Here we show that in PDAC cell lines with amplified MYC, MondoA is required for viability, facilitating proliferation while suppressing apoptosis in vitro and in vivo. Transcriptional and genomic profiling demonstrates that loss of MondoA leads to altered expression of direct MondoA targets as well as MYC target genes and is accompanied by shifts in genomic occupancy of MYC, MNT, and the MondoA paralog ChREBP. This altered genomic binding by MYC network members is associated with transcriptional perturbation of multiple metabolic and stress pathways, as well as global changes in N6-methyladenosine modification (m6A) of mRNA. MondoA inhibition disrupts coordination between MYC network members and the Integrated Stress Response (ISR), resulting in decreased translation of ATF4 mRNA, discordant gene regulation of shared targets of MYC and ATF4 and, ultimately, apoptosis. Re-establishing ATF4 protein expression rescues the diminished viability due to loss of MondoA expression or activity, providing direct evidence of a link between deregulated MYC and the transcriptional machinery of the ISR. Lastly, we find that small-molecule inhibition of MondoA is lethal in a subset of PDAC cell lines, including patient-derived organoids, suggesting that the ability to target MYC via chemical inhibition of MondoA transcriptional activity may have broad efficacy.

Significance StatementThis report investigates mechanisms underlying the dependence of MYC-amplified pancreatic cancer cells on the MYC network member MondoA which, as a heterodimer with MLX, is a nutrient-sensing transcription factor. We show this dependency is linked to genomic crosstalk between MYC, components of the proximal MYC network, and the master regulator of the integrated stress response, ATF4. Moreover, we find that small molecule inhibitors of MondoA-MLX transcriptional activity abrogate survival of MYC-amplified PDAC lines and patient derived organoids. The significance of this work relates to its focus on a unique vulnerability intrinsic to MYC, an oncogenic driver associated with a wide range of cancers, which is considered to be "undruggable".
]]></description>
<dc:creator>Ramsey, E. L.</dc:creator>
<dc:creator>Dobersch, S.</dc:creator>
<dc:creator>Freie, B.</dc:creator>
<dc:creator>Hong, N. H.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Kugel, S.</dc:creator>
<dc:creator>Eisenman, R. N.</dc:creator>
<dc:creator>Carroll, P. A.</dc:creator>
<dc:date>2025-09-08</dc:date>
<dc:identifier>doi:10.1101/2025.09.03.674106</dc:identifier>
<dc:title><![CDATA[MondoA mediates transcriptional coordination between the MYC network and the integrated stress response in pancreatic ductal adenocarcinoma]]></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/10.1101/2025.09.05.671932v1?rss=1">
<title>
<![CDATA[
BIOPRINTING OF MICRODISSECTED TUMOR CUBOIDS IN HYDROGELS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.05.671932v1?rss=1</link>
<description><![CDATA[
Microscale tumor models made from microdissected tumors that retain much of the original human tumor microenvironment (TME) are emerging as an alternative to preclinical animal models. We have introduced a drug testing approach that utilizes regularly-cut, cuboidal-shaped microdissected tissues, or "cuboids," as a way to maximize creation of microtissues from scarce biopsy materials. However, microtissues (e.g., cuboids, organoids, spheroids, etc.) can be difficult to place in precise locations, especially in applications that require their culture in hydrogels. Here, using cuboids from mouse tumor models, we demonstrate a simple bioprinting strategy for precise placement and immobilization of cuboids in hydrogel. We use a commercial bioprinter to bioprint-containing hydrogel into arrays of small hydrogel dots containing cuboids, or "cuboid dots," either onto a Transwell insert or into traps on a microplate. The hydrogel serves to immobilize the cuboids in place and provides a matrix to support cuboid viability. We demonstrate proof-of-concept applications for cancer drug testing and for protein profiling analysis. This approach will enable interface of cuboids with other devices, such as on top of a sensor or in a microfluidic platform. Furthermore, this automated process of dispensing and localizing cuboids (or other microtissue formats such as spheroids or organoids) could further their application to drug discovery and personalized medicine.

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]]></description>
<dc:creator>Bansal, A. M.</dc:creator>
<dc:creator>Horowitz, L.</dc:creator>
<dc:creator>Yeung, M.</dc:creator>
<dc:creator>Gujral, T. M.</dc:creator>
<dc:creator>Folch, A.</dc:creator>
<dc:date>2025-09-10</dc:date>
<dc:identifier>doi:10.1101/2025.09.05.671932</dc:identifier>
<dc:title><![CDATA[BIOPRINTING OF MICRODISSECTED TUMOR CUBOIDS IN HYDROGELS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.05.674437v1?rss=1">
<title>
<![CDATA[
Identification of a broad and potent V3 glycan site bNAb targeting anN332gp120 glycan-independent epitope 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.05.674437v1?rss=1</link>
<description><![CDATA[
Broadly neutralizing antibodies (bNAbs) against HIV-1 can suppress viremia in vivo and inform vaccine development. Here, we characterized 007, a V3 glycan site bNAb exhibiting high levels of antiviral activity against multiclade pseudovirus panels1-3 (GeoMean IC50 = 0.012 {micro}g/mL, breadth = 69%, 217 virus strains) by targeting a N332gp120 glycan-independent V3 epitope, a site of Env vulnerability to which only weakly neutralizing antibodies had previously been identified. Functional analyses demonstrated distinct binding and neutralization profiles compared to classical V3 glycan site bNAbs. A 007 Fab-Env cryo-EM structure revealed contacts with the V3 324GD/NIR327 motif and interactions with N156gp120 and N301gp120 glycans. In contrast to classical V3 bNAbs, 007 binding to Env does not depend on the N332gp120 glycan, rendering it resistant to common escape mutations. Structures of 007 IgG-Env trimer complexes showed two Env trimers crosslinked by three bivalent IgGs, and bivalent 007 IgG was up to [~]300-fold more potent than monovalent 007 IgG heterodimer, suggesting a role for avidity in potent neutralization. Finally, in HIV-1ADA-infected humanized mice, 007 caused transient decline of viremia and overcame classical V3 escape mutations, highlighting 007s potential for HIV-1 prevention, therapy, functional cure, and vaccine design.
]]></description>
<dc:creator>Gieselmann, L.</dc:creator>
<dc:creator>DeLaitsch, A. T.</dc:creator>
<dc:creator>Rohde, M.</dc:creator>
<dc:creator>Radford, C.</dc:creator>
<dc:creator>Worczinski, J.</dc:creator>
<dc:creator>Momot, A.</dc:creator>
<dc:creator>Ahmadov, E.</dc:creator>
<dc:creator>Burger, J. A.</dc:creator>
<dc:creator>Havenar-Daughton, C.</dc:creator>
<dc:creator>Deshpande, S.</dc:creator>
<dc:creator>Giovannoni, F.</dc:creator>
<dc:creator>Corti, D.</dc:creator>
<dc:creator>Kreer, C.</dc:creator>
<dc:creator>Ercanoglu, M. S.</dc:creator>
<dc:creator>Schommers, P.</dc:creator>
<dc:creator>Georgiev, I.</dc:creator>
<dc:creator>West, A. P.</dc:creator>
<dc:creator>Knuefer, J.</dc:creator>
<dc:creator>Stumpf, R.</dc:creator>
<dc:creator>Kroidl, A.</dc:creator>
<dc:creator>Geldmacher, C.</dc:creator>
<dc:creator>Maganga, L.</dc:creator>
<dc:creator>William, W.</dc:creator>
<dc:creator>Ntinginya, N. E.</dc:creator>
<dc:creator>Hoelscher, M.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Wei, Q.</dc:creator>
<dc:creator>Renfrow, M.</dc:creator>
<dc:creator>Green, T. J.</dc:creator>
<dc:creator>Novak, J.</dc:creator>
<dc:creator>van Gils, M.</dc:creator>
<dc:creator>Gristick, H. B.</dc:creator>
<dc:creator>Gruell, H.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Bjorkman, P.</dc:creator>
<dc:creator>Klein, F.</dc:creator>
<dc:date>2025-09-10</dc:date>
<dc:identifier>doi:10.1101/2025.09.05.674437</dc:identifier>
<dc:title><![CDATA[Identification of a broad and potent V3 glycan site bNAb targeting anN332gp120 glycan-independent epitope]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.05.674562v1?rss=1">
<title>
<![CDATA[
Genotoxic antibody-drug conjugates combined with Bcl-xL inhibitors enhance therapeutic efficacy in metastatic castration-resistant prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.05.674562v1?rss=1</link>
<description><![CDATA[
Metastatic castration-resistant prostate cancer (mCRPC) is an aggressive subtype of prostate cancer (PC) without curative treatments. Antibody-drug conjugates (ADCs) emerged as promising cancer therapeutics that selectively deliver cytotoxic agents (payloads) to the tumors. Although ADCs have been successfully applied in the treatment of hematological and solid tumors, ADC monotherapy has not demonstrated durable responses in mCRPC and the mechanisms of PC resistance to ADCs have not been thoroughly investigated. Our study aimed to improve ADC efficacy using a new integrated approach for custom ADC design and multiplexing. To nominate rational combinations of ADC targets and ADC payloads, we (1) examined protein co-expression of three clinically relevant surface antigens-- B7 homolog 3 (B7-H3), prostate specific membrane antigen (PSMA), and six-transmembrane epithelial antigen of prostate-1 (STEAP1)--in a series of human mCRPC samples and (2) screened established ADC payloads and their combinations in mCRPC cell lines with different phenotypes. We identified synergistic interactions between DNA-damaging payloads and Bcl-xL inhibitor A-1331852 as well as their coordinated induction of the intrinsic apoptosis pathway. The functional relevance of isolated p53 loss and impaired PC responses to three genotoxic ADCs (B7-H3-seco-DUBA, PSMA-SG3249, and STEAP1-DXd) and their combinations with A-1331852 was established using genetic knockout models. Lastly, we found enhanced in vivo antitumor activity in mCRPC by combining the clinically relevant agents B7-H3-seco-DUBA (vobramitamab duocarmazine) and A-1331852. Collectively, our findings provide rationale for the development of ADC therapies combining genotoxic payloads with Bcl-xL inhibitors for mCRPC.

SignificanceB7-H3, PSMA, and STEAP1 targeted ADC therapies combining genotoxic payloads with Bcl-xL inhibitors induce p53-dependant apoptotic cell death in mCRPC, providing a clinically viable strategy for the treatment of advanced prostate cancer.
]]></description>
<dc:creator>Semenova, G.</dc:creator>
<dc:creator>Frank, S.</dc:creator>
<dc:creator>Dumpit, R.</dc:creator>
<dc:creator>Han, W.</dc:creator>
<dc:creator>Coleman, I.</dc:creator>
<dc:creator>Gulati, R.</dc:creator>
<dc:creator>Morrissey, C.</dc:creator>
<dc:creator>Haffner, M.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:date>2025-09-11</dc:date>
<dc:identifier>doi:10.1101/2025.09.05.674562</dc:identifier>
<dc:title><![CDATA[Genotoxic antibody-drug conjugates combined with Bcl-xL inhibitors enhance therapeutic efficacy in metastatic castration-resistant prostate cancer]]></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/10.1101/2025.09.12.675951v1?rss=1">
<title>
<![CDATA[
Quantifying HLA transcripts by genotype in chimeric mixtures at single-cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.12.675951v1?rss=1</link>
<description><![CDATA[
Gene products from the highly variable major histocompatibility locus, including HLA, are essential for self-recognition and immune surveillance of malignancy. Following allogeneic hematopoietic cell transplantation (alloHCT), genetic and epigenetic alterations in HLA can drive disease recurrence, making precise HLA assessment critical for determining future therapy. However, current methods lack the sensitivity to quantify HLA transcripts at the single-cell level, limiting their clinical utility. We introduce scrHLA-typing, a novel technique that accurately identifies and quantifies HLA transcripts in single cells using long-read sequencing. When applied to samples from patients with post-transplant relapse, scrHLA-typing successfully detected HLA allele-specific expression, across a range of levels of donor-recipient chimerism, at clinically actionable levels. By characterizing allele expression in residual leukemia cells, our assay identified differences in expression patterns among patients. This capability highlights scrHLA-typings potential to improve risk stratification and guide the selection of appropriate salvage therapies, enhancing personalized treatment strategies after relapse.
]]></description>
<dc:creator>Kanaan, S. B.</dc:creator>
<dc:creator>Underwood, J. G.</dc:creator>
<dc:creator>Green Gladden, R.</dc:creator>
<dc:creator>Fan, E.</dc:creator>
<dc:creator>Bhise, S. S.</dc:creator>
<dc:creator>Thakar, M. S.</dc:creator>
<dc:creator>Jaeger-Ruckstuhl, C. A.</dc:creator>
<dc:creator>Stevens, J.</dc:creator>
<dc:creator>Gray, A. N.</dc:creator>
<dc:creator>Riddell, S. R.</dc:creator>
<dc:creator>Bleakley, M.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Furlan, S. N.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.12.675951</dc:identifier>
<dc:title><![CDATA[Quantifying HLA transcripts by genotype in chimeric mixtures at single-cell resolution]]></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/10.1101/2025.09.13.675985v1?rss=1">
<title>
<![CDATA[
Identification of protective human monoclonal antibodies using a K18 hACE2 transgenic mouse SARS-CoV-2 challenge model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.13.675985v1?rss=1</link>
<description><![CDATA[
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), uses human angiotensin converting enzyme 2 (hACE2) as its obligate receptor for cell entry. The K18 hACE2 transgenic mouse line, which expresses hACE2 under control of the human keratin 18 (K18) promoter, is used as an animal model for the study of COVID-19 pathogenesis. Here, we evaluate this model in the screening of human monoclonal antibody (hmAb) therapies against SARS-CoV-2. We included 206 hmAbs from the Coronavirus Immunotherapeutic Consortium Database (CoVIC-DB) and identified many that protected against a lethal challenge with the virus. Our data showed that mouse weight change from day 5 onward highly correlated with survival. Many of the protective hmAb candidates we identified also showed strong viral neutralization and spike protein (SP) binding when measured in vitro; however, in many cases, in vitro assays failed to identify protective hmAbs, suggesting that the mouse model may capture characteristics of the hmAbs that other methods cannot. Our findings demonstrate the relevance of including in vivo models for the characterization of therapeutics against SARS-CoV-2, as these improve both accuracy and expediency in the screening process.
]]></description>
<dc:creator>Borate, B.</dc:creator>
<dc:creator>Pino, P. A.</dc:creator>
<dc:creator>Hicks, A.</dc:creator>
<dc:creator>Garcia, J. I.</dc:creator>
<dc:creator>Garcia-Vilanova, A.</dc:creator>
<dc:creator>Ye, C.</dc:creator>
<dc:creator>Park, J.-G.</dc:creator>
<dc:creator>Maingot, B.</dc:creator>
<dc:creator>Rodriguez, O.</dc:creator>
<dc:creator>Saveria, T.</dc:creator>
<dc:creator>Holman, D.</dc:creator>
<dc:creator>Schendel, S. L.</dc:creator>
<dc:creator>Saphire, E. O.</dc:creator>
<dc:creator>Martinez-Sobrido, L.</dc:creator>
<dc:creator>Torrelles, J. B.</dc:creator>
<dc:creator>Fiore-Gartland, A.</dc:creator>
<dc:date>2025-09-19</dc:date>
<dc:identifier>doi:10.1101/2025.09.13.675985</dc:identifier>
<dc:title><![CDATA[Identification of protective human monoclonal antibodies using a K18 hACE2 transgenic mouse SARS-CoV-2 challenge model]]></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/10.1101/2025.09.22.677165v1?rss=1">
<title>
<![CDATA[
A connexin 43 targeting peptide prevents blood vessel neointima formation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.22.677165v1?rss=1</link>
<description><![CDATA[
The gap junction protein connexin 43 (Cx43) is associated with human pathological vascular smooth muscle cell (SMC) proliferation and neointima formation. We previously identified mitogen-activated protein kinase (MAPK) phosphorylation of Cx43 results in binding with the cell cycle protein cyclin E, facilitating neointima formation in mice. However, the specific nature of these interactions and their relevance to human disease have not been elucidated. Using an ex vivo human saphenous vein model of neointima formation, we identified increased MAPK-phosphorylated Cx43 and cyclin E in explant tissues. We used peptide arrays to define a cyclin E-Cx43 binding region and generated  CycliCx, a stearate-linked Cx43 phospho-mimetic peptide. In human coronary artery SMC, CycliCx inhibits platelet-derived growth factor-BB (PDGF-{beta})-induced changes in Cx43 trafficking and interactions with cyclin E, and stimulation of proliferation. RNAseq analysis identified CycliCx significantly inhibits PDGF-{beta}-induced proliferative pathways in SMC by limiting PDGF-{beta}-induced early G1/S phase cell cycle progression transcripts. Finally, we show CycliCx limits neointima formation in mice in vivo and in ex vivo human saphenous vein explants. Our data provide strong evidence for selective targeting of Cx43 as a viable therapeutic strategy for preventing neointimal formation in humans.
]]></description>
<dc:creator>Sedovy, M. W.</dc:creator>
<dc:creator>Renton, M. C.</dc:creator>
<dc:creator>Leng, X.</dc:creator>
<dc:creator>Iqbal, F.</dc:creator>
<dc:creator>Leaf, M.</dc:creator>
<dc:creator>Roberts, K.</dc:creator>
<dc:creator>Joshi, R. C.</dc:creator>
<dc:creator>Malek, A.</dc:creator>
<dc:creator>Dennison, C.</dc:creator>
<dc:creator>Best, A. K.</dc:creator>
<dc:creator>Baker, J. W.</dc:creator>
<dc:creator>Joseph, M.</dc:creator>
<dc:creator>Lampe, P. D.</dc:creator>
<dc:creator>Isakson, B. E.</dc:creator>
<dc:creator>Johnstone, S. R.</dc:creator>
<dc:date>2025-09-24</dc:date>
<dc:identifier>doi:10.1101/2025.09.22.677165</dc:identifier>
<dc:title><![CDATA[A connexin 43 targeting peptide prevents blood vessel neointima formation]]></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/10.1101/2025.09.24.678258v1?rss=1">
<title>
<![CDATA[
HRDPath: An Explainable Multi-Model Deep Learning Architecture for Predicting Homologous Recombination Deficiency from Histopathology Images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.24.678258v1?rss=1</link>
<description><![CDATA[
Homologous recombination deficiency (HRD) is a critical biomarker for guiding treatment decisions in high-grade serous tubo-ovarian carcinoma (HGSOC), a cancer with few reliable biomarkers. However, existing genomic-based tests for HRD are variable, expensive, and time-consuming. To this end, we developed HRDPath, a novel patient-level deep learning architecture that combines the strengths of two complementary models with a multi-task design, to predict genomically derived HRD status from whole slide images in HGSOC. HRDPath was comprehensively validated across three datasets and benchmarked against leading deep learning models. It achieved an AUC of 0.846, surpassing previously reported H&E-based HRD prediction results for HGSOC images by 0.09, and for the first time, reporting a specificity of 0.938, where accuracy significantly increased when multiple slides per patient were used. Our proposed patient-level approach and interpretability pipeline enhance model trustworthiness and reveal important clinical and biological insights into HRD-positive cancers, highlighting the associated morphological and pathological changes at the cellular and tissue levels. HRDPath is a potentially accessible and scalable digital biomarker that could improve ovarian cancer diagnosis and therapy selection.
]]></description>
<dc:creator>Wu, C.-i.</dc:creator>
<dc:creator>Banda, K.</dc:creator>
<dc:creator>Arora, R.</dc:creator>
<dc:creator>Soong, T. R.</dc:creator>
<dc:creator>Radke, M.</dc:creator>
<dc:creator>Dillon, M.</dc:creator>
<dc:creator>Nik-Zainal, S.</dc:creator>
<dc:creator>Swisher, E.</dc:creator>
<dc:creator>Sailem, H.</dc:creator>
<dc:date>2025-09-26</dc:date>
<dc:identifier>doi:10.1101/2025.09.24.678258</dc:identifier>
<dc:title><![CDATA[HRDPath: An Explainable Multi-Model Deep Learning Architecture for Predicting Homologous Recombination Deficiency from Histopathology Images]]></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/10.1101/2025.09.25.678525v1?rss=1">
<title>
<![CDATA[
Plasma nirmatrelvir and molnupiravir concentrations required for inhibition of SARS CoV-2 replication differ between rhesus macaques and humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.25.678525v1?rss=1</link>
<description><![CDATA[
Early during the COVID-19 pandemic, non-human primate (NHP) infection models emerged as highly useful tools for preclinical screening of antiviral drugs. However, it is uncertain whether NHP models can be used to precisely inform optimal dosing in humans. We previously established and validated mathematical models which were fit to SARS-CoV-2 viral loads from human clinical trials. These models identified that plasma drug concentrations required to inhibit viral replication by 50% in humans (in vivo EC50) differ substantially from in vitro EC50 estimates in cell culture systems. Here we apply models to sequential viral load data from SARS-CoV-2 infected rhesus macaques (RM) that were untreated or treated with nirmatrelvir/ritonavir, molnupiravir, or both drugs. We identify that equivalent plasma drug concentrations correspond to greater antiviral potency in lungs compared to nasal passages for nirmatrelvir and molnupiravir. Average nirmatrelvir antiviral efficacy in RM (30% in nasal passages and 46% in lungs) was estimated to be less than in humans (82%) due to shorter plasma drug half-life. Molnupiravir efficacy in RM (95% in nasal and 99% in lungs) is estimated to be similar to efficacy in humans against omicron variants. Our model estimates that 10-fold higher plasma nirmatelvir concentrations are needed in humans versus RM to achieve 50% reduction in viral replication, whereas 20-fold lower plasma molnupiravir concentrations are needed. Our results suggest that dose optimization in humans based on modeling of NHP viral loads is limited by drug-specific differences in pharmacokinetic, pharmacodynamic and virologic profiles, and that data from human phase 1 and 2 trials is better suited for this task.
]]></description>
<dc:creator>Avila-Ponce de Leon, U.</dc:creator>
<dc:creator>Esmaeili, S.</dc:creator>
<dc:creator>Owens, K.</dc:creator>
<dc:creator>Schiffer, J. T.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.09.25.678525</dc:identifier>
<dc:title><![CDATA[Plasma nirmatrelvir and molnupiravir concentrations required for inhibition of SARS CoV-2 replication differ between rhesus macaques and humans]]></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/10.1101/2025.09.29.677401v1?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/10.1101/2025.09.30.679620v1?rss=1">
<title>
<![CDATA[
Ontogeny and functional potential of founding dendritic cells in the developing lung 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.30.679620v1?rss=1</link>
<description><![CDATA[
Lung development begins in utero and reaches full maturity post birth. Dendritic cells (DC) play a key role in immune regulation in lungs. However, comprehensive exploration of DCs in these immature lungs has not been performed. Here we explored DCs from fetal to newborn mouse lungs phenotypically, ontogenetically, transcriptomically and functionally and found two DC subsets, resembling adult cDC1 and cDC2, but with key differences. Phenotypically, fetal-cDC1 lacks the classical-DC1 (cDC1) marker XCR1, while the fetal-cDC2 express both cDC-associated genes as well as monocyte-derived DC genes. Both DC subsets wane as lungs enter the alveolar stage, giving way to the more familiar adult cDC1 and cDC2. Both fetal-cDC1 and fetal-cDC2 derive from ED14.5 fetal liver Macrophage Dendritic Progenitors, not from monocytes or classic Precursor-cDC (Pre-cDC), indicating a unique ontogeny of first DCs in developing mouse lungs. Together we provide the first in depth exploration of first DCs in developing lungs.
]]></description>
<dc:creator>Headley, M. B.</dc:creator>
<dc:creator>Soysa, R.</dc:creator>
<dc:creator>Abideen, S.</dc:creator>
<dc:creator>Reyes, V. Z.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.09.30.679620</dc:identifier>
<dc:title><![CDATA[Ontogeny and functional potential of founding dendritic cells in the developing lung]]></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/10.1101/2025.09.30.679621v1?rss=1">
<title>
<![CDATA[
Population-level migration modeling of North American birds through data integration with BirdFlow 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.30.679621v1?rss=1</link>
<description><![CDATA[
BackgroundAccurate information on population-level movements of migratory animals is essential for understanding migration and for designing effective conservation strategies in a changing world. Yet such information remains scarce for most migratory species due to the effort and expense needed to collect data across their full distribution ranges. BirdFlow is a probabilistic modeling framework that infers population-level movements from weekly species distribution maps produced by the participatory science project eBird. However, BirdFlow models have only been tuned for a handful of species using high-resolution individual tracking data, which is not available for most migratory species.

MethodsHere, we introduce a general tuning and evaluation framework for BirdFlow that enables the first large-scale integration of distributional and individual-level data to infer animal movement across continents and hundreds of migratory species, eliminating reliance on any single individual-tracking data source. By generalizing the BirdFlow model parametrization, we enable tuning and validation using multiple complementary data sources, including GPS tracks, banding recoveries, and radio telemetry data from the Motus Wildlife Tracking System. We investigate the efficacy of this approach by (1) investigating predictive performance compared to null models; (2) validating the biological plausibility of BirdFlow models by comparing movement properties such as route straightness, number of stopovers, and migration speed between model-generated routes and real movement tracks; and (3) comparing the performance of models tuned on species-specific movement data to models tuned using hyperparameters transferred from other species.

ResultsOur results show that BirdFlow models produced by the new tuning framework achieve biologically realistic performance, even for prediction horizons of thousands of kilometers and several months. When species-specific data are unavailable, models can still be tuned using data from other phylogenetically adjacent species to achieve improved performance.

ConclusionsBy integrating eBird Status & Trends abundance surfaces with data from banding recaptures, radio telemetry, and GPS tracking, we scale BirdFlow model to 153 North American migratory species, representing the first collection of continental-scale population-level movement and forecasting models. Species-specific tuning improves population-level movement forecasts, while taxonomically informed hyperparameter transfer supports the modeling of data-limited species. Overall, our work offers a foundation for more accurate predictions across hundreds of species for research in ecology and conservation, disease surveillance, aviation, and public outreach.
]]></description>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Slager, D. L.</dc:creator>
<dc:creator>Plunkett, E.</dc:creator>
<dc:creator>Fuentes, M.</dc:creator>
<dc:creator>Deng, Y.</dc:creator>
<dc:creator>Mackenzie, S. A.</dc:creator>
<dc:creator>Berrigan, L. E.</dc:creator>
<dc:creator>Fink, D.</dc:creator>
<dc:creator>Sheldon, D.</dc:creator>
<dc:creator>Van Doren, B. M.</dc:creator>
<dc:creator>Dokter, A. M.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.09.30.679621</dc:identifier>
<dc:title><![CDATA[Population-level migration modeling of North American birds through data integration with BirdFlow]]></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/10.1101/2025.09.30.679684v1?rss=1">
<title>
<![CDATA[
spCorr: flexible and scalable inference of spatially varying correlation in spatial transcriptomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.30.679684v1?rss=1</link>
<description><![CDATA[
Spatial transcriptomics has transformed our ability to explore gene expression within its tissue context, enabling us to dissect subtle yet biologically significant variations in situ. While numerous computational methods have been proposed for detecting Spatially Varying Genes (SVGs) expression by modeling each individual gene separately, much less effort has been devoted to understanding how correlations between genes change across space. Such Spatially Varying Correlations (SVCs) are critical for understanding biological processes such as gene regulatory mechanisms shaped by local tissue environments, yet existing tools remain limited for this task. To address this gap, we present spCorr, a flexible and scalable regression framework for studying SVCs. spCorr provides interpretable, spot-level estimates of gene correlation and detects gene pairs whose correlations vary across locations or between tissue domains. Through extensive simulations and real-data analyses, we show that spCorr achieves high detection power, reliably controls the False Discovery Rate (FDR), and is computationally efficient. Importantly, spCorr reveals biologically meaningful correlation patterns that highlight fine-scale tissue structures, gene module functions, and region-specific interactions, offering new opportunities to study coordinated gene regulation in spatial transcriptomics.
]]></description>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Yin, Y.</dc:creator>
<dc:creator>Robson, P.</dc:creator>
<dc:creator>Li, J. Y. H.</dc:creator>
<dc:creator>Li, J. J.</dc:creator>
<dc:creator>Song, D.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.09.30.679684</dc:identifier>
<dc:title><![CDATA[spCorr: flexible and scalable inference of spatially varying correlation in spatial transcriptomics]]></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/10.1101/2025.10.01.679929v1?rss=1">
<title>
<![CDATA[
De novo design of RNA and nucleoprotein complexes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.01.679929v1?rss=1</link>
<description><![CDATA[
Nucleic acids fold into sequence-dependent tertiary structures and carry out diverse biological functions, much like proteins. However, while considerable advances have been made in the de novo design of protein structure and function, the same has not yet been achieved for RNA tertiary structures of similar intricacy. Here, we describe a generative diffusion framework, RFDpoly, for generalized de novo biopolymer (RNA, DNA and protein) design, and use it to create diverse and designable RNA structures. We design RNA structures with novel folds and experimentally validate them using a combination of chemical footprinting (SHAPE-seq) and electron microscopy. We further use this approach to design protein-nucleic acid assemblies; the crystal structure of one such design is nearly identical to the design model. This work demonstrates that the principles of structure-based de novo protein design can be extended to nucleic acids, opening the door to creating a wide range of new RNA structures and protein-nucleic acid complexes.
]]></description>
<dc:creator>Favor, A. H.</dc:creator>
<dc:creator>Quijano, R.</dc:creator>
<dc:creator>Chernova, E.</dc:creator>
<dc:creator>Kubaney, A.</dc:creator>
<dc:creator>Weidle, C.</dc:creator>
<dc:creator>Esler, M. A.</dc:creator>
<dc:creator>McHugh, L.</dc:creator>
<dc:creator>Carr, A.</dc:creator>
<dc:creator>Hsia, Y.</dc:creator>
<dc:creator>Juergens, D.</dc:creator>
<dc:creator>Carr, K. D.</dc:creator>
<dc:creator>Kim, P. T.</dc:creator>
<dc:creator>Politanska, Y.</dc:creator>
<dc:creator>Sehgal, E.</dc:creator>
<dc:creator>Kwon, P. S.</dc:creator>
<dc:creator>Pecoraro, R. J.</dc:creator>
<dc:creator>Glasscock, C.</dc:creator>
<dc:creator>Borst, A. J.</dc:creator>
<dc:creator>DiMaio, F.</dc:creator>
<dc:creator>Stoddard, B. L.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679929</dc:identifier>
<dc:title><![CDATA[De novo design of RNA and nucleoprotein complexes]]></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/10.1101/2025.10.03.679858v1?rss=1">
<title>
<![CDATA[
Re-infection with SARS-CoV-2 is associated with increased antibody breadth and potency against diverse sarbecovirus strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.03.679858v1?rss=1</link>
<description><![CDATA[
The ease with which emerging SARS-CoV-2 variants escape neutralizing antibodies limits protection afforded by a prior exposure, be it infection or vaccination. While rare, broadly neutralizing antibodies with activity towards diverse sarbecoviruses have been detected in convalescent serum. Motivated by findings that plasma responses show increased neutralization breadth and potency with continued antigen exposure, we isolated monoclonal antibodies (mAbs) after a SARS-CoV-2 re-infection and compared them to those isolated one year prior, after the first breakthrough infection. Among clonal lineage members identified at both time points, mAbs from the later time point showed improved neutralization potency and breadth. One mAb isolated after re-infection, C68.490, targets a conserved region in the receptor binding domain core and shows remarkable activity not only against SARS-CoV-2 variants, but also diverse sarbecoviruses from more distant clades present in animal reservoirs. These findings suggest that a focus on individuals with diverse and repeated antigen exposure could lead to identification of antibodies with therapeutic utility not just towards current and future SARS-CoV-2 variants, but also distant sarbecoviruses in the event of a future spillover.
]]></description>
<dc:creator>Lilly, M.</dc:creator>
<dc:creator>Ruiz, F.</dc:creator>
<dc:creator>Foreman, W.</dc:creator>
<dc:creator>Chohan, V.</dc:creator>
<dc:creator>Guenthoer, J.</dc:creator>
<dc:creator>Depierreux, D.</dc:creator>
<dc:creator>Baharani, V.</dc:creator>
<dc:creator>Ralph, D.</dc:creator>
<dc:creator>Harteloo, A.</dc:creator>
<dc:creator>Chu, H.</dc:creator>
<dc:creator>Bieniasz, P. D.</dc:creator>
<dc:creator>Starr, T.</dc:creator>
<dc:creator>Overbaugh, J.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.03.679858</dc:identifier>
<dc:title><![CDATA[Re-infection with SARS-CoV-2 is associated with increased antibody breadth and potency against diverse sarbecovirus strains]]></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/10.1101/2025.10.03.680278v1?rss=1">
<title>
<![CDATA[
Characterization of quiescent subpopulations and proliferative compartments in glioblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.03.680278v1?rss=1</link>
<description><![CDATA[
Glioblastoma (GBM) quiescent (Q) cell populations are hypothesized to contain cancer stem-like cells (CSC) that drive tumor growth, cellular heterogeneity, and recurrence. However, GBM tumors do not neatly resolve into developmental hierarchies and Q stem-like activities are difficult to assess. Here, we evaluated tumor Q subpopulations in patient-derived GBM xenograft tumors using live cell reporters, DNA label retention assays, and single cell genomics. Compared to adult neural stems cells (NSCs), GBM Q populations contain hybrid transcriptional states composed of networks found in both dormant and activated adult NSCs, resulting in constitutive expression of key Q egress transcription factors and their targets (e.g., AP-1 and CCND1/2). As a result, even the longest Q-residing cells ([~]12 days) in xenograft tumors continuously cycle and fail to enter dormant Q states. We provide evidence and hypothesize that transient Q states in primary tumors arise as part of distinct proliferative compartments rather than deterministic developmental hierarchies driven by CSC activity. We further speculate that increases in basal translation rates drive Q instability in GBM tumors.
]]></description>
<dc:creator>Mihalas, A.</dc:creator>
<dc:creator>Mitchell, K.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>O'Connor, S.</dc:creator>
<dc:creator>Patel, A.</dc:creator>
<dc:creator>Plaisier, C. L.</dc:creator>
<dc:creator>PADDISON, P.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.03.680278</dc:identifier>
<dc:title><![CDATA[Characterization of quiescent subpopulations and proliferative compartments in glioblastoma]]></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/10.1101/2025.10.08.680531v1?rss=1">
<title>
<![CDATA[
A searchable database of publications using fluorescent probes and flow cytometry to study antigen-specific B cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.08.680531v1?rss=1</link>
<description><![CDATA[
The study of antigen-specific B cells has resulted in important advances in all fields of immunology, the development of experimentally and/or clinically useful antibodies, and as a starting point for rationally designed vaccine antigens. A key innovation allowing for widespread study of antigen-specific B cells was the development of fluorescent antigen probes for use with flow cytometry. Initially these studies were mostly focused upon B cells specific for a variety of model antigens, but over the past decade focus has shifted towards the study of B cells specific for antigens from pathogens such as SARS-CoV-2, HIV, and Influenza virus. Importantly however, these types of approaches have been used for hundreds of different antigens and could be used for thousands more. Unfortunately, studies of B cells specific for an antigen of interest are not easily searchable on current publication databases since these assays are often a small portion of a larger publication. To overcome this, we built a searchable database of studies analyzing antigen-specific B cells by flow cytometry using fluorescent antigen probes that is located at www.immunology.virginia.edu/Taylor/Bcell/Database.php. Using this database, we assessed the number of publications per year revealing rapid growth in the use of this approach in recent years. While much of this rapid growth was focused upon the assessment of B cells specific for SARS-CoV-2, HIV-1, or Influenza virus, studies assessing B cells specific for hundreds of different antigens derived from numerous microbes, animals, plants, or other sources can be found in the database. Combined, the antigen-specific B cell database was built to facilitate identification of studies assessing these cells and for analysis of the field as a whole.



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]]></description>
<dc:creator>Palani, S.</dc:creator>
<dc:creator>Kong, A. X.</dc:creator>
<dc:creator>Buetow, A.</dc:creator>
<dc:creator>Yousif, A. S.</dc:creator>
<dc:creator>Say, A.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Biswas, K.</dc:creator>
<dc:creator>Blaszczyk, J.</dc:creator>
<dc:creator>Rodrigues-Jesus, M. J.</dc:creator>
<dc:creator>Malave Ramos, D. R.</dc:creator>
<dc:creator>Senthilkumar, V.</dc:creator>
<dc:creator>Chappell, S.</dc:creator>
<dc:creator>Koenig, J. F.</dc:creator>
<dc:creator>Taylor, J. J.</dc:creator>
<dc:date>2025-10-09</dc:date>
<dc:identifier>doi:10.1101/2025.10.08.680531</dc:identifier>
<dc:title><![CDATA[A searchable database of publications using fluorescent probes and flow cytometry to study antigen-specific B cells]]></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/10.1101/2025.10.09.681477v1?rss=1">
<title>
<![CDATA[
RNA regulates repeat-associated non-AUG (RAN) translation initiation in C9orf72 FTD/ALS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.09.681477v1?rss=1</link>
<description><![CDATA[
Repeat-associated non-AUG (RAN) translation synthesizes protein in the absence of a cognate AUG start codon 1,2In frontotemporal dementia and amyotrophic lateral sclerosis, a GGGGCC (G4C2) repeat expansion in an intron of C9orf72 leads to synthesis of neurotoxic dipeptide-repeat proteins, underscoring the need to understand the mechanism of C9orf72 RAN translation1-5. RNA sequence and structure have been implicated, but how they direct C9orf72 RAN translation, particularly the rate-limiting, multi-step initiation phase, remains unclear6-10. We applied single-molecule biophysics to a reconstituted human translation initiation system and tracked fluorescently labeled ribosomes and initiation factors in real time. We show that RNA G4C2 repeats and sequence context alter initiation factor dynamics after ribosomal scanning, generating a kinetic bottleneck in the commitment to initiate at a near-cognate CUG start codon. Our model of C9orf72 RAN translation provides a mechanistic framework for how repeat expansions change underlying translation dynamics and may be broadly relevant to other disorders that involve RAN translation.
]]></description>
<dc:creator>Grosely, R.</dc:creator>
<dc:creator>Yamada-Hunter, S.</dc:creator>
<dc:creator>Puglisi, A.</dc:creator>
<dc:creator>Palo, M. Z.</dc:creator>
<dc:creator>Alvarado, C.</dc:creator>
<dc:creator>Lapointe, C. P.</dc:creator>
<dc:creator>Gitler, A. D.</dc:creator>
<dc:creator>Puglisi, J. D.</dc:creator>
<dc:date>2025-10-09</dc:date>
<dc:identifier>doi:10.1101/2025.10.09.681477</dc:identifier>
<dc:title><![CDATA[RNA regulates repeat-associated non-AUG (RAN) translation initiation in C9orf72 FTD/ALS]]></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/10.1101/2025.10.10.681694v1?rss=1">
<title>
<![CDATA[
Obesity Impairs the Antitumor Activity of CAR-T Cells in Triple-Negative Breast Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.10.681694v1?rss=1</link>
<description><![CDATA[
BackgroundWe have reported that chimeric antigen receptor (CAR) T cells targeting B7-H3 (B7-H3.CAR) are effective in a preclinical model of triple negative breast cancer (TNBC), and have initiated a Phase I study to assess safety and efficacy. However, heterogeneous antigen expression and immunosuppressive tumor microenvironments (TME) remain roadblocks for effective CAR-T cell therapy. In particular, obesity represents a negative prognostic factor in TNBC partly due to chronic inflammation and impaired adaptive immune responses. Hence, we sought to determine if obesity can affect the antitumor activity of B7-H3.CAR-T cells.

MethodsWe used qPCR and western blotting to determine if cytokines associated with obesity affect B7-H3 expression in TNBC cell lines. Furthermore, we used shRNA to suppress B7-H3 expression in a syngeneic orthotopic E0771 tumor model and measured tumor growth in control and diet-induced obese (DIO) mice. Finally, we evaluated the antitumor effects of B7-H3.CAR-T cells in both control and DIO mice orthotopically engrafted with the E0771 tumor cell line. Immune profiling was conducted using flow cytometry.

ResultsObesity-related inflammatory cytokines promote B7-H3 expression in human and murine TNBC cells in vitro and B7-H3 expression correlates with tumor aggressiveness in vivo. CAR-T cells obtained from control or DIO mice were equally cytotoxic in vitro but activated T cells and B7-H3.CAR-T cells obtained from DIO mice show transcriptomic changes (enriched Tox2, Prdm1, Batf) and impaired glycolytic capacity, respectively. Finally, we demonstrated that obesity impairs CAR-T cell antitumor effects and durability of response in vivo with a near complete loss of memory formation.

ConclusionsHere we identified a correlation between B7-H3 expression, obesity, and rate of tumor growth in TNBC. Furthermore, we showed that obesity constrains both the ability of B7-H3.CAR-T cells to control tumor growth and to elicit durable immunological memory. Taken together, these data identify obesity as an underappreciated and potent modulator of CAR-T cell functionality.

What is already knownB7-H3 protein is upregulated in many human malignancies including TNBC and is often associated with worsened outcomes. B7-H3.CAR-T cells show promise in preclinical models of TNBC and entered clinical translation.

What this study addsThis study identifies a previously unknown correlation between obesity, B7-H3 expression, and rate of tumor growth in TNBC. Furthermore, our preclinical model of B7-H3.CAR-T cell therapy demonstrates that obesity negatively affects the antitumor activity of B7-H3.CAR-T cells in TNBC.

How this study affects other research/practiceThis study highlights obesity as an understudied and critically important covariate for adoptive T-cell therapy and demonstrates important links between systemic metabolism and antigen expression. This work paves the way for future mechanistic and translational research into how obesity impacts CAR-T cell functionality.
]]></description>
<dc:creator>Malian, H. M.</dc:creator>
<dc:creator>Marnata Pellegry, C.</dc:creator>
<dc:creator>Oh, H. M.</dc:creator>
<dc:creator>Glenny, E. M.</dc:creator>
<dc:creator>Ho, A. N.</dc:creator>
<dc:creator>Dotti, G.</dc:creator>
<dc:creator>Hursting, S. D.</dc:creator>
<dc:creator>Coleman, M. F.</dc:creator>
<dc:date>2025-10-13</dc:date>
<dc:identifier>doi:10.1101/2025.10.10.681694</dc:identifier>
<dc:title><![CDATA[Obesity Impairs the Antitumor Activity of CAR-T Cells in Triple-Negative Breast Cancer]]></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/10.1101/2025.10.10.681725v1?rss=1">
<title>
<![CDATA[
A telomere-to-telomere map of somatic mutation burden and functional impact in cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.10.681725v1?rss=1</link>
<description><![CDATA[
Oncogenesis involves widespread genetic and epigenetic alterations, yet the full spectrum of somatic variation genome-wide remains unresolved. We generated a near-telomere-to-telomere (T2T) diploid assembly of a donor paired with deep short- and long-read sequencing of their melanoma. This revealed that 16% of somatic variants occur in sequences absent from GRCh38, with satellite repeats acting as hotspots for UV-induced damage due to sequence-intrinsic mutability and inefficient repair. Centromere kinetochore domains emerged as focal sites of structural, genetic, and epigenetic variation, leading to remodeling of centromere kinetochore binding domains during tumor evolution. Single-molecule telomere reconstructions uncovered cycles of attrition, deletion, and telomerase-mediated extension that shape cancer telomeres. Finally, diploid chromatin maps exposed that copy number alterations and epimutations, rather than point mutations, predominate in rewiring cancer regulatory programs. These findings define the full landscape of a cancers somatic variation and their functional impact, establishing a blueprint for T2T studies of mosaicism.
]]></description>
<dc:creator>Sohn, M.-H.</dc:creator>
<dc:creator>Dubocanin, D.</dc:creator>
<dc:creator>Vollger, M. R.</dc:creator>
<dc:creator>Kwon, Y.</dc:creator>
<dc:creator>Minkina, A.</dc:creator>
<dc:creator>Munson, K. M.</dc:creator>
<dc:creator>Hart, S. F.</dc:creator>
<dc:creator>Ranchalis, J. E.</dc:creator>
<dc:creator>Parmalee, N. L.</dc:creator>
<dc:creator>Sedeno-Cortes, A. E.</dc:creator>
<dc:creator>Ou, J.</dc:creator>
<dc:creator>Au, N. Y.</dc:creator>
<dc:creator>Bohaczuk, S.</dc:creator>
<dc:creator>Carroll, B.</dc:creator>
<dc:creator>Frazar, C. D.</dc:creator>
<dc:creator>Harvey, W. T.</dc:creator>
<dc:creator>Hoekzema, K.</dc:creator>
<dc:creator>Huang, M.-F.</dc:creator>
<dc:creator>Jacques, C. N.</dc:creator>
<dc:creator>Jensen, D. M.</dc:creator>
<dc:creator>Kolar, J. T.</dc:creator>
<dc:creator>Lee, R.</dc:creator>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Loy, K.</dc:creator>
<dc:creator>Mack, T.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Pham, M. M.</dc:creator>
<dc:creator>Ryke, E.</dc:creator>
<dc:creator>Smith, J. D.</dc:creator>
<dc:creator>Sutherlin, L.</dc:creator>
<dc:creator>Swanson, E. G.</dc:creator>
<dc:creator>Weiss, J. M.</dc:creator>
<dc:creator>SMaHT Assembly Working Group,</dc:creator>
<dc:creator>Carvalho, C.</dc:creator>
<dc:creator>Coorens, T. H.</dc:creator>
<dc:creator>Harris, K.</dc:creator>
<dc:creator>Wei, C.-L.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:creator>Altemose, N.</dc:creator>
<dc:creator>Bennett, J. T.</dc:creator>
<dc:creator>Stergachis, A. B.</dc:creator>
<dc:date>2025-10-13</dc:date>
<dc:identifier>doi:10.1101/2025.10.10.681725</dc:identifier>
<dc:title><![CDATA[A telomere-to-telomere map of somatic mutation burden and functional impact in cancer]]></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/10.1101/2025.10.08.680958v1?rss=1">
<title>
<![CDATA[
Moving Beyond Binary Biomarkers: Machine Learning Model Resolves Concurrent and Molecularly Heterogeneous Mismatch Repair and Homologous Recombination Deficiencies in Prostate Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.08.680958v1?rss=1</link>
<description><![CDATA[
Current DNA damage repair (DDR) biomarkers employ binary classifications that fail to capture the molecular complexity of tumors with concurrent repair deficiencies. We used genomics analysis to stratify 672 metastatic prostate cancer patients into 11 DDR subgroups, identifying 51 molecular signatures with weighted roles in class identity. We identified a tumor-mutational-burden very-high subset, characterized by 19 mutations/Mb or more, as a molecularly distinct group characterized by preserved genomic integrity and enhanced immunogenicity. Critically, 2.3 percent of tumors exhibited concurrent TMB-High and HRR mutant phenotypes, while 1.5 percent harbored MMR bi-allelic loss without MMRd (mismatch-repair-deficiency) signatures. Clinical validation in 130 patients demonstrated superior immunotherapy responses in tumors with very high TMB levels. We developed CHIMERA DDR, a probabilistic machine learning tool that integrates these 51 genomic features using a nested Random Forest architecture to infer seven clinically relevant DDR subgroups. After negating model overfit concerns, CHIMERA-DDR showed exceptional classification performance (AUCs 0.919-0.999) to accurately detect MMRd and HRR mutant molecular subtypes with or without concurrent DDR deficiencies, resolving admixed phenotypes to enable precision therapeutic stratification beyond binary methods.
]]></description>
<dc:creator>Sharma, K.</dc:creator>
<dc:creator>Wilson, D. A.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Haider, S.</dc:creator>
<dc:creator>Bhatlapenumarthi, V.</dc:creator>
<dc:creator>Boiarsky, D.</dc:creator>
<dc:creator>Cipriaso, J. M.</dc:creator>
<dc:creator>Yermakov, L.</dc:creator>
<dc:creator>Dutta, R.</dc:creator>
<dc:creator>Coleman, I. M.</dc:creator>
<dc:creator>Bankhead, A.</dc:creator>
<dc:creator>Jadhav, S. K.</dc:creator>
<dc:creator>Neupane, B.</dc:creator>
<dc:creator>Taylor, B. W.</dc:creator>
<dc:creator>Morrissey, C.</dc:creator>
<dc:creator>Schweitzer, M. T.</dc:creator>
<dc:creator>Montgomery, R. B.</dc:creator>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Nevalainen, M. T.</dc:creator>
<dc:creator>Nelson, A. A.</dc:creator>
<dc:creator>Antonarakis, E. S.</dc:creator>
<dc:creator>Pilea, P. G.</dc:creator>
<dc:creator>Berchuk, J. E.</dc:creator>
<dc:creator>Zarrabi, K. K.</dc:creator>
<dc:creator>Pritchard, C.</dc:creator>
<dc:creator>Kothari, A.</dc:creator>
<dc:creator>Chen, H.-Z.</dc:creator>
<dc:creator>George, B.</dc:creator>
<dc:creator>Kurzrock, R.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Banerjee, A.</dc:creator>
<dc:creator>Auer, P. L.</dc:creator>
<dc:creator>Kilari, D.</dc:creator>
<dc:creator>De Sarkar, N.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.08.680958</dc:identifier>
<dc:title><![CDATA[Moving Beyond Binary Biomarkers: Machine Learning Model Resolves Concurrent and Molecularly Heterogeneous Mismatch Repair and Homologous Recombination Deficiencies in Prostate Cancer]]></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/10.1101/2025.10.13.681989v1?rss=1">
<title>
<![CDATA[
Pervasive suppressors halt the spread of selfish Segregation Distorter in a natural population 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.13.681989v1?rss=1</link>
<description><![CDATA[
Meiotic drivers are selfish genetic elements that subvert Mendelian inheritance to increase their own transmission, yet they are typically found at low frequencies across natural populations. The factors that limit their spread remain unclear. To investigate this paradox, we studied the Segregation Distorter (SD) system, a selfish coadapted gene complex in Drosophila melanogaster. SD biases its transmission by killing sperm carrying a homologous chromosome bearing a target locus, Responder (Rsp), which appear as satellite repeats. Such selfish killing impairs male fertility and imposes selective pressure on the host genome to evolve resistance, either by deleting Rsp copies or acquiring unlinked suppressors. To characterize the spectrum of Rsp alleles and the frequency of segregating suppressors, we surveyed 90 strains from the Drosophila Genome Reference Panel. Rather than loss of Rsp, we found that over half of the strains (52/90) harbor suppressors located on the X chromosome or autosomes, but not the Y chromosome. The widespread presence of strong suppressors limited the resolution of our genome-wide association mapping; however, recombination analysis identified a strong X-linked suppressor to a [~]300 kb interval on the chromosome. Together, our findings suggest that pervasive, multilocus suppression constrains the spread of SD in natural populations.
]]></description>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Handler, T.</dc:creator>
<dc:creator>Fuda, N.</dc:creator>
<dc:creator>Pascua, D.</dc:creator>
<dc:creator>Mouton, T.</dc:creator>
<dc:creator>Larracuente, A. M.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.13.681989</dc:identifier>
<dc:title><![CDATA[Pervasive suppressors halt the spread of selfish Segregation Distorter in a natural population]]></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/10.1101/2025.10.13.682212v1?rss=1">
<title>
<![CDATA[
Nascent CUT&Tag captures transcription factor binding after chromatin duplication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.13.682212v1?rss=1</link>
<description><![CDATA[
DNA replication strips off all chromatin proteins, which must be reassembled behind the replication fork. To track chromatin reassembly on newly synthesized DNA, we developed Nascent CUT&Tag, a chromatin profiling method that directly measures transcription factor (TF) binding on nascent chromatin. We tracked the recovery of GAGA factor (GAF) and Pleiohomeotic (PHO) in Drosophila Kc167 cells. Whereas both TFs are evicted following passage of the replication fork, GAF recovers on newly synthesized DNA over a minutes-to-hours range, whereas PHO generally requires hours to fully reestablish binding. Early recovering GAF peaks are characterized by shorter GAF motifs and are associated with functions related to cell cycle progression. Conversely, late recovering peaks are characterized by longer, degenerate GAF motifs and are associated with developmental functions. GAF recovery on newly synthesized DNA requires chromatin remodeling by Brahma Associated Factor (BAF), implying that nucleosome turnover is critical to fully reestablish GAF binding.
]]></description>
<dc:creator>Wooten, M.</dc:creator>
<dc:creator>Nguyen, K.</dc:creator>
<dc:creator>Takushi, B. N.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:date>2025-10-15</dc:date>
<dc:identifier>doi:10.1101/2025.10.13.682212</dc:identifier>
<dc:title><![CDATA[Nascent CUT&Tag captures transcription factor binding after chromatin duplication]]></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/10.1101/2025.10.15.682664v1?rss=1">
<title>
<![CDATA[
Functional and antigenic constraints on the Nipah virus fusion protein 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.15.682664v1?rss=1</link>
<description><![CDATA[
Nipah virus is a highly pathogenic virus in the family Paramyxoviridae that utilizes two distinct surface glycoproteins to infect cells. The receptor-binding protein (RBP) binds host receptors whereas the fusion protein (F) merges viral and host membranes. Here, we use non-replicative pseudoviruses to safely measure the effects of all F single amino-acid residue mutations on its cell entry function and neutralization by monoclonal antibodies. We compare mutational tolerance in F with previous experimental measurements for RBP and show that F is much more functionally constrained than the RBP. We also identify mutationally intolerant sites on the F trimer surface and core that are critical for proper function, and describe mutations that are candidates for stabilizing F in the prefusion conformation for vaccine design. We quantify how F mutations affect neutralization by six monoclonal antibodies, and show that the magnitude of mutational effects on neutralization varies among antibodies. Our measurements of mutational effects on Nipah virus F predict the ability of the antibodies to neutralize the related Hendra virus. Overall, our work defines the functional and antigenic constraints on the F protein from an important zoonotic virus.

ImportanceNipah virus sporadically spills over into humans, where it is often fatal. The Nipah fusion (F) protein is necessary for infection, and is a target for vaccines and antibody therapies. To better understand the constraints on this protein, we experimentally measured how [~]8,500 single amino-acid mutations to F affected its function using pseudoviruses that enable the safe study of protein mutants without the generation of actual replicative virus. We examined the effects of these mutations in the context of structural data and publicly available Nipah virus sequences to characterize the constraints that shape F protein evolution. This work has implications for understanding paramyxovirus fusion proteins, and informs the development of vaccines and monoclonal antibody therapies.
]]></description>
<dc:creator>Larsen, B. B.</dc:creator>
<dc:creator>Harari, S.</dc:creator>
<dc:creator>Gen, R.</dc:creator>
<dc:creator>Stewart, C.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2025-10-15</dc:date>
<dc:identifier>doi:10.1101/2025.10.15.682664</dc:identifier>
<dc:title><![CDATA[Functional and antigenic constraints on the Nipah virus fusion protein]]></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/10.1101/2025.10.17.683141v1?rss=1">
<title>
<![CDATA[
Evidence of latency reshapes our understanding of Ebola virus reservoir dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.17.683141v1?rss=1</link>
<description><![CDATA[
The reservoir of Ebola virus (EBOV) has remained a mystery since the first recorded zoonotic epidemic in 1976. While recent outbreaks have revealed much about the epidemiological dynamics that sustain human-to-human transmission, the mechanisms by which the virus persists between epidemics are unknown. Observations of extreme EBOV evolutionary rate heterogeneity in humans suggest latency is an important, yet untested, factor in persistence between epidemics. We here explicitly model latency with a novel phylogenetic approach to characterise the natural history of EBOV and, by extension, its reservoir. We find evidence that EBOV undergoes extended periods (i.e., possibly decades) of quiescence in the reservoir. Accounting for these dynamics significantly changes our understanding of EBOVs evolutionary past and its geographic spread through Central Africa.
]]></description>
<dc:creator>McCrone, J. T.</dc:creator>
<dc:creator>Baele, G.</dc:creator>
<dc:creator>Omah, I. F.</dc:creator>
<dc:creator>Kinganda-Lusamaki, E.</dc:creator>
<dc:creator>Brew, J. A.</dc:creator>
<dc:creator>Carvalho, L. M.</dc:creator>
<dc:creator>Dudas, G.</dc:creator>
<dc:creator>Mbala-Kingebeni, P.</dc:creator>
<dc:creator>Suchard, M. A.</dc:creator>
<dc:creator>Rambaut, A.</dc:creator>
<dc:date>2025-10-18</dc:date>
<dc:identifier>doi:10.1101/2025.10.17.683141</dc:identifier>
<dc:title><![CDATA[Evidence of latency reshapes our understanding of Ebola virus reservoir dynamics]]></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/10.1101/2025.10.20.682949v1?rss=1">
<title>
<![CDATA[
Cross-Species Morphology Learning Enables Nucleic Acid-Independent Detection of Live Mutant Blood Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.20.682949v1?rss=1</link>
<description><![CDATA[
In both neonates and adults, the presence of malignancy-associated mutations in peripheral blood (PB) correlates with an elevated risk of future neoplastic transformation, with certain mutations, such as KMT2A rearrangements, exhibiting near-complete penetrance. If feasible, pre-malignant screening could enable early intervention and even disease prevention. However, nucleic acid sequencing- and hybridization-based mutation detection have limited cost-efficiency, constraining their use in screening. Here, we introduce a computer vision platform that can identify mutant cells in fresh PB samples that carry KMT2A-MLLT3 (a frequent mutation in pediatric and adult leukemias and detectable in newborn blood samples) or JAK2V617F (a frequent mutation in myeloproliferative neoplasms and clonal hematopoiesis). This is achieved by high-throughput single-cell imaging and mutation detection by machine learning (ML)-powered morphology recognition. The ML models were developed by cross-species learning of conserved features between mutant cells from mouse genetic models and from human samples, enabling a cost-effective approach for detecting mutations in live blood cells. This platform holds promise for pre-malignant screening in asymptomatic neonates and adults with KMT2A-MLLT3 or JAK2V617Fmutation and is potentially generalizable to the detection other malignancy-associate mutations. Our platform provides a novel single-cell morphological data modality that complements existing single-cell genomics.
]]></description>
<dc:creator>Khan, S. A.</dc:creator>
<dc:creator>Faerber, D.</dc:creator>
<dc:creator>Kirkey, D.</dc:creator>
<dc:creator>Raffel, S.</dc:creator>
<dc:creator>Hadland, B.</dc:creator>
<dc:creator>Deininger, M.</dc:creator>
<dc:creator>Buettner, F.</dc:creator>
<dc:creator>Zhao, H. G.</dc:creator>
<dc:date>2025-10-21</dc:date>
<dc:identifier>doi:10.1101/2025.10.20.682949</dc:identifier>
<dc:title><![CDATA[Cross-Species Morphology Learning Enables Nucleic Acid-Independent Detection of Live Mutant Blood Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.21.683652v1?rss=1">
<title>
<![CDATA[
Separating selection from mutation in antibody language models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.21.683652v1?rss=1</link>
<description><![CDATA[
Antibodies are encoded by nucleotide sequences that are generated by V(D)J recombination and evolve according to mutation and selection processes. Existing antibody language models, however, focus exclusively on antibodies as strings of amino acids and are fitted using standard language modeling objectives such as masked or autoregressive prediction. In this paper, we first show that fitting models using this objective implicitly incorporates nucleotide-level mutation processes as part of the protein language model, which degrades performance when predicting effects of mutations on functional properties of antibodies. To address this limitation, we devise a new framework: a Deep Amino acid Selection Model (DASM) that learns the selection effects of amino-acid mutations while explicitly factoring out the nucleotide-level mutation process. By fitting selection as a separate term from the mutation process, the DASM exclusively quantifies functional effects: effects that change some aspect of the function of the antibody. This factorization leads to substantially improved performance on standard functional benchmarks. Moreover, our model is an order of magnitude smaller and multiple orders of magnitude faster to evaluate than existing approaches, as well as being readily interpretable.
]]></description>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:creator>Dumm, W.</dc:creator>
<dc:creator>Sung, K.</dc:creator>
<dc:creator>Johnson, M. M.</dc:creator>
<dc:creator>Rich, D.</dc:creator>
<dc:creator>Starr, T.</dc:creator>
<dc:creator>Song, Y. S.</dc:creator>
<dc:creator>Fukuyama, J.</dc:creator>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:date>2025-10-22</dc:date>
<dc:identifier>doi:10.1101/2025.10.21.683652</dc:identifier>
<dc:title><![CDATA[Separating selection from mutation in antibody language models]]></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/10.1101/2025.10.26.684319v1?rss=1">
<title>
<![CDATA[
Inflammatory Biomarkers of Asymptomatic and Symptomatic Tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.26.684319v1?rss=1</link>
<description><![CDATA[
A large proportion of individuals with tuberculosis (TB) are asymptomatic. The biological and inflammatory underpinnings of asymptomatic TB are unknown and may differ from symptomatic TB. We characterised blood transcriptomic and proteomic profiles in South African community screening vs. health facility-based triage cohorts. Asymptomatic TB shared core transcriptomic and proteomic features with symptomatic TB, including upregulation of innate, interferon and inflammatory pathways and downregulation of T and B cell pathways. Integration of transcriptomic and proteomic data from asymptomatic TB individuals identified two distinct sub-clusters characterized by higher or lower bacterial burden, blood IFN-{gamma} responses, BMI, and chest radiographic abnormalities, suggesting different disease severity. We identified a new blood transcriptomic signature of asymptomatic TB. However, diagnostic performance of transcriptomic and proteomic markers was weaker for asymptomatic TB than symptomatic TB, suggesting that policy development for community-based, asymptomatic TB screening should not adopt biomarkers developed for symptomatic TB triage without further optimization.
]]></description>
<dc:creator>Awany, D.</dc:creator>
<dc:creator>Ariefdien, D.</dc:creator>
<dc:creator>Mendelsohn, S.</dc:creator>
<dc:creator>Rozot, V.</dc:creator>
<dc:creator>Mulenga, H.</dc:creator>
<dc:creator>Nyangu, S.</dc:creator>
<dc:creator>Tameris, M.</dc:creator>
<dc:creator>Moloantoa, T.</dc:creator>
<dc:creator>Katona, A.</dc:creator>
<dc:creator>Maruri, F.</dc:creator>
<dc:creator>Noor, F.</dc:creator>
<dc:creator>Panchiar, R.</dc:creator>
<dc:creator>Hlongwane, K.</dc:creator>
<dc:creator>Stanley, K.</dc:creator>
<dc:creator>Van der Heijden, Y.</dc:creator>
<dc:creator>Hadley, K.</dc:creator>
<dc:creator>Fiore-Gartland, A.</dc:creator>
<dc:creator>Innes, C.</dc:creator>
<dc:creator>Brumskine, W.</dc:creator>
<dc:creator>Dheda, K.</dc:creator>
<dc:creator>Jaumdally, S.</dc:creator>
<dc:creator>Perumal, T.</dc:creator>
<dc:creator>Martinson, N.</dc:creator>
<dc:creator>Leslie, A.</dc:creator>
<dc:creator>Fourie, B.</dc:creator>
<dc:creator>Hiemstra, A.</dc:creator>
<dc:creator>Malherbe, S.</dc:creator>
<dc:creator>Walzl, G.</dc:creator>
<dc:creator>Naidoo, K.</dc:creator>
<dc:creator>Churchyard, G.</dc:creator>
<dc:creator>Chegou, N.</dc:creator>
<dc:creator>Sterling, T.</dc:creator>
<dc:creator>Hatherill, M.</dc:creator>
<dc:creator>Scriba, T. J.</dc:creator>
<dc:date>2025-10-29</dc:date>
<dc:identifier>doi:10.1101/2025.10.26.684319</dc:identifier>
<dc:title><![CDATA[Inflammatory Biomarkers of Asymptomatic and Symptomatic Tuberculosis]]></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/10.1101/2025.10.29.685411v1?rss=1">
<title>
<![CDATA[
VRC01 Selects Rare HIV Escape Mutations After Acquisition in Antibody-Mediated Prevention Trials 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.29.685411v1?rss=1</link>
<description><![CDATA[
Broadly neutralizing antibodies (bnAbs) show promise in HIV prevention, yet viral escape remains a challenge. In the Antibody Mediated Prevention (AMP) trials, the CD4 binding site (CD4bs) bNAb VRC01 blocked acquisition by VRC01-sensitive strains. However, its influence on viral evolution post-acquisition is not fully understood. Here we analyzed >12,000 HIV env sequences from 47 participants from the AMP trials, identifying VRC01-mediated de novo escape mutations in 8 of 26 VRC01-treated participants but none in 21 placebo participants. These mutations were found at very low frequency (<1%) in global viruses. Escape mutations, primarily located in the Loop-D and {beta}23/V5 regions of Env, conferred cross-resistance to several CD4bs bnAbs, while more potent CD4bs bnAbs like N6 and 1-18 largely retained their activity. Our findings demonstrate that prophylactic VRC01 can select for viral escape after infection, underscoring the need for next-generation bnAbs with improved breadth and potency to enhance durability and efficacy of antibody-based HIV prevention.
]]></description>
<dc:creator>Williamson, C.</dc:creator>
<dc:creator>Curry, L.</dc:creator>
<dc:creator>Mkhize, N. N.</dc:creator>
<dc:creator>Giorgi, E. E.</dc:creator>
<dc:creator>Magaret, C. A.</dc:creator>
<dc:creator>Lambson, B. E.</dc:creator>
<dc:creator>Bhebhe, S.</dc:creator>
<dc:creator>Kaldine, H.</dc:creator>
<dc:creator>Moyo-Gwete, T.</dc:creator>
<dc:creator>Rolland, M.</dc:creator>
<dc:creator>Rossenkhan, R.</dc:creator>
<dc:creator>Garcia, N. M. G.</dc:creator>
<dc:creator>Moodley, C.</dc:creator>
<dc:creator>Yssel, A.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Marsden, A. A.</dc:creator>
<dc:creator>Reeves, D. B.</dc:creator>
<dc:creator>Mayer, B. T.</dc:creator>
<dc:creator>Bumgarner, R. E.</dc:creator>
<dc:creator>Beaume, N.</dc:creator>
<dc:creator>Westfall, D. H.</dc:creator>
<dc:creator>Juraska, M.</dc:creator>
<dc:creator>DeCamp, A. C.</dc:creator>
<dc:creator>Murrell, H. G.</dc:creator>
<dc:creator>Bai, H.</dc:creator>
<dc:creator>Deng, W.</dc:creator>
<dc:creator>Pankow, A.</dc:creator>
<dc:creator>Bhattacharya, T.</dc:creator>
<dc:creator>York, T.</dc:creator>
<dc:creator>Ndabambi, N.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Gwashu-Nyangiri, A.</dc:creator>
<dc:creator>Thebus, R.</dc:creator>
<dc:creator>Cohen, P.</dc:creator>
<dc:creator>Murrell, B.</dc:creator>
<dc:creator>Karuna, S.</dc:creator>
<dc:creator>Hural, J.</dc:creator>
<dc:creator>Mgodi, N.</dc:creator>
<dc:creator>Edupuganti, S.</dc:creator>
<dc:creator>Morris, L.</dc:creator>
<dc:creator>Montefiori, D.</dc:creator>
<dc:creator>McElrath, M. J.</dc:creator>
<dc:creator>Cohen,</dc:creator>
<dc:date>2025-10-29</dc:date>
<dc:identifier>doi:10.1101/2025.10.29.685411</dc:identifier>
<dc:title><![CDATA[VRC01 Selects Rare HIV Escape Mutations After Acquisition in Antibody-Mediated Prevention Trials]]></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/10.1101/2025.10.29.685473v1?rss=1">
<title>
<![CDATA[
Characterization of Cancer-Reactive T Cells and Neoantigen-specific T Cell Receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.29.685473v1?rss=1</link>
<description><![CDATA[
Not all tumor-infiltrating T cells are cancer-reactive (CR). Several studies have reported gene expression signatures associated with CR-T cells. To integrate these findings, we developed a computational workflow, CAT (Cancer-Associated T cells), which harmonizes existing CR-T cell signatures and applies them to identify CR-T cells across an atlas of one million T cells. Our findings reveal that the abundance of CR-T cells varies across cancer types and that baseline levels of CR-T cells predict patients responses to immunotherapy. In parallel, we established a high-throughput computational platform, Neo-TCR, for systematic screening of neoantigen-specific TCRs and their cognate neoantigens. The efficacy of Neo-TCR is validated by cross-validation studies and replications in two independent datasets. Together, our findings suggest a new direction for developing biomarkers for cancer detection and monitoring: integrating CR gene expression signatures with neoantigen-specific TCRs.
]]></description>
<dc:creator>Sun, W.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Bleakley, M.</dc:creator>
<dc:date>2025-10-30</dc:date>
<dc:identifier>doi:10.1101/2025.10.29.685473</dc:identifier>
<dc:title><![CDATA[Characterization of Cancer-Reactive T Cells and Neoantigen-specific T Cell Receptors]]></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/10.1101/2025.10.29.685337v1?rss=1">
<title>
<![CDATA[
larch: mapping the parsimony-optimal landscape of trees for directed exploration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.29.685337v1?rss=1</link>
<description><![CDATA[
Phylogenetic inference algorithms for large data sets typically return a single tree. However, there are often many optimal trees, especially when sequence data is closely related. We develop a compact representation of large collections of maximally parsimonious histories--trees with mutations mapped onto tree edges. Our C++ implementation, larch, leverages this representation for a highly parallel search algorithm. The storage component uses our history DAG structure to compactly represent large families of optimal trees. The search algorithm integrates this storage with matOptimize for rapid tree optimization; the DAG structure allows us to accept thousands of conflicting tree rearrangements in parallel. The integration enables a new type of tree search: one that systematically maps out the collection of good trees, enabling moves that are directed away from the current set of optimal trees to cross valleys and increase the diversity of the set of optimal trees. It is able to identify more parsimonious trees than are found by other methods. We find diverse optimality landscapes for viral datasets, including many distinct plateaux. We also find that our implementation produces similar results whether using a variety of single starting trees or an ensemble of starting trees, indicating effective global optimization.
]]></description>
<dc:creator>Barker, M.</dc:creator>
<dc:creator>Milanov, O.</dc:creator>
<dc:creator>Dumm, W.</dc:creator>
<dc:creator>Rich, D.</dc:creator>
<dc:creator>Turakhia, Y.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2025-10-31</dc:date>
<dc:identifier>doi:10.1101/2025.10.29.685337</dc:identifier>
<dc:title><![CDATA[larch: mapping the parsimony-optimal landscape of trees for directed exploration]]></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/10.1101/2025.10.30.685502v1?rss=1">
<title>
<![CDATA[
ADAPTIVE MULTI-SCALE GRAPH TRANSFORMER FRAMEWORK FORHISTOPATHOLOGICAL IMAGES 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.30.685502v1?rss=1</link>
<description><![CDATA[
Whole slide images (WSIs) contain hierarchical information from cellular to tissue architecture but their gigapixel scale poses major memory and computational challenges. Existing multi-scale graph and transformer models capture complex WSI features effectively but struggle with efficiency. We propose an Adaptive Multi-Scale Graph Transformer (AMGT) for WSI classification that addresses this limitation through two key modules: a Self-Guided Token Aggregation (SGTA) mechanism that fuses multi-resolution features to reduce redundancy, and a Prototypical Transformer (PT) that groups similar tokens into phenotype-representative prototypes with linear complexity. This design preserves essential spatial and semantic information, substantially lowering memory cost and improving interpretability by prototypical learning. AMGT achieves superior performance and efficiency, outperforming state-of-the-art models by 1.8% and 5.3% AUC on high-grade ovarian cancer and Camelyon16 datasets, respectively. These results demonstrate AMGTs capacity for scalable, interpretable multi-scale representation learning.
]]></description>
<dc:creator>Wu, C.-i.</dc:creator>
<dc:creator>Banda, K.</dc:creator>
<dc:creator>Swisher, E.</dc:creator>
<dc:creator>Sailem, H.</dc:creator>
<dc:date>2025-10-31</dc:date>
<dc:identifier>doi:10.1101/2025.10.30.685502</dc:identifier>
<dc:title><![CDATA[ADAPTIVE MULTI-SCALE GRAPH TRANSFORMER FRAMEWORK FORHISTOPATHOLOGICAL IMAGES]]></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/10.1101/2025.10.30.685582v1?rss=1">
<title>
<![CDATA[
Inhibition of farnesyltransferase activity diminishes hematopoietic stem cell ex vivo expansion ability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.30.685582v1?rss=1</link>
<description><![CDATA[
ABSTRACT/SUMMARYThe rationale underlying this work is that elucidating the contribution of farnesyltransferase activity to hematopoietic stem cell expansion ex vivo will provide knowledge needed to better support hematopoietic stem cell expansion techniques for clinical applications that rely on this approach, like hematopoietic cell transplants and gene therapy. We discovered that pharmacological inhibition of farnesyltransferase activity with lonafarnib substantially diminished the ex vivo expansion potential of human and mouse hematopoietic stem cells, highlighting that hematopoietic stem cells rely on isoprenoids for their ex vivo maintenance.
]]></description>
<dc:creator>Newman, C.</dc:creator>
<dc:creator>Alves, I. M.</dc:creator>
<dc:creator>Hagen, M. W.</dc:creator>
<dc:creator>Wellington, R.</dc:creator>
<dc:creator>Hadland, B. K.</dc:creator>
<dc:creator>Quintana, A. M.</dc:creator>
<dc:creator>Termini, C. M.</dc:creator>
<dc:date>2025-10-31</dc:date>
<dc:identifier>doi:10.1101/2025.10.30.685582</dc:identifier>
<dc:title><![CDATA[Inhibition of farnesyltransferase activity diminishes hematopoietic stem cell ex vivo expansion ability]]></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/10.1101/2025.11.07.687077v1?rss=1">
<title>
<![CDATA[
Depletion of microenvironmental syndecan-2 impairs hematopoietic stem cell self-renewal and cytokine responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.07.687077v1?rss=1</link>
<description><![CDATA[
Syndecan-2 is a heparan sulfate proteoglycan highly enriched on murine bone marrow hematopoietic stem cells (HSCs) compared to terminally differentiated hematopoietic cells. Syndecan-2 binds growth factors via its heparan sulfate glycosaminoglycan chains to coordinate cell signaling. Knockdown of syndecan-2 reduces HSC self-renewal ability and promotes cell cycling via Cdkn1c. In this study, we analyzed the function of syndecan-2 expressed by bone marrow niche cells in hematopoiesis and HSC self-renewal. We determined that syndecan-2 is highly expressed by bone marrow mesenchymal stromal cells and moderately expressed by endothelial cells. To test the function of niche-expressed syndecan-2 in hematopoiesis, we generated transgenic mice depleted of Sdc2 in Lepr-targeted mesenchymal stromal cells (Sdc2{Delta}MSC mice) or Cdh5-targeted endothelial cells (Sdc2{Delta}EC mice). Loss of syndecan-2 from endothelial or mesenchymal stromal cells did not change bone marrow HSC frequencies or numbers. However, depletion of syndecan-2 from Lepr-targeted mesenchymal stromal cells, but not Cdh5-targeted endothelial cells, diminishes HSC self-renewal ability analyzed by competitive transplants into lethally irradiated mice. Ex vivo studies further show that HSCs co-cultured with HS-5 stromal cells depleted of SDC2 exhaust more rapidly than HSCs cultured with control HS-5 cells. Single-cell RNA sequencing analyses reveal that the depletion of Sdc2 from mesenchymal stromal cells significantly remodels the HSC transcriptome by enriching for pathways associated with excessive growth factor signaling. Together, our findings suggest that HSC self-renewal is supported by cell-extrinsic mechanisms enacted by syndecan-2 from the MSC niche, highlighting the importance of the niche proteoglycome in HSC functions.

KEY POINTSO_LIThe heparan sulfate proteoglycan syndecan-2 expressed by mesenchymal stromal cells but not endothelial cells regulates HSC self-renewal
C_LIO_LIDepletion of syndecan-2 from Lepr-targeted mesenchymal stromal cells remodels the hematopoietic stem cell transcriptional landscape
C_LI
]]></description>
<dc:creator>Hagen, M. W.</dc:creator>
<dc:creator>Setiawan, N. J.</dc:creator>
<dc:creator>Wellington, R.</dc:creator>
<dc:creator>Woodruff, K. A.</dc:creator>
<dc:creator>Newman, C.</dc:creator>
<dc:creator>Billings, T. M.</dc:creator>
<dc:creator>Nazzaro, M. N.</dc:creator>
<dc:creator>Hadland, B.</dc:creator>
<dc:creator>Termini, C. M.</dc:creator>
<dc:date>2025-11-09</dc:date>
<dc:identifier>doi:10.1101/2025.11.07.687077</dc:identifier>
<dc:title><![CDATA[Depletion of microenvironmental syndecan-2 impairs hematopoietic stem cell self-renewal and cytokine responses]]></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/10.1101/2025.11.08.687354v1?rss=1">
<title>
<![CDATA[
Hidden Markov Models Detect Recombination and Ancestry of SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.08.687354v1?rss=1</link>
<description><![CDATA[
When individuals are co-infected with distinct SARS-CoV-2 lineages, homologous recombination can generate mosaic genomes carrying mutations from both parental lineages. A variety of methods exist to detect recombinant sequences and their parental lineages in surveillance-scale datasets comprised of millions of SARS-CoV-2 genomes. However, these methods often rely on user-specified parameters, such as the probability a recombination breakpoint occurs between adjacent positions on the query sequence. In this study, we devise a hidden Markov model that detects recombinant SARS-CoV-2 sequences and identifies their parental lineages within a test set of sequences. Our method does not depend on user-specified parameters and can accommodate de novo mutations on the query sequence that are not present in the predicted parental lineages. To achieve this, we use maximum likelihood to estimate parameters that characterize the transition and emission probabilities in our hidden Markov model. Applying our method to 440,307 SARS-CoV-2 sequences sampled in England between September 2020 and March 2024, we detect 7,619 recombinant sequences corresponding to 1.73% (95% CI: [1.69%, 1.77%]) of all sampled sequences. We observe a positive association between the proportion of query sequences detected as recombinant in each week and community SARS-CoV-2 prevalence. This is consistent with higher prevalence increasing the risk of co-infection by distinct lineages and promoting the emergence of recombinant sequences. Finally, we observe localized clusters of recombination breakpoints within spike and in intergenic regions.
]]></description>
<dc:creator>Masaki, N.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:date>2025-11-10</dc:date>
<dc:identifier>doi:10.1101/2025.11.08.687354</dc:identifier>
<dc:title><![CDATA[Hidden Markov Models Detect Recombination and Ancestry of SARS-CoV-2]]></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/10.1101/2025.11.10.687711v1?rss=1">
<title>
<![CDATA[
Superabundant microRNAs are transcribed from human rDNA spacer promoters insulated by CTCF 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.10.687711v1?rss=1</link>
<description><![CDATA[
MicroRNAs are [~]22-nucleotide RNAs processed from primary transcripts and exported from the nucleus to repress gene expression by base-pairing to mRNAs. We find that the highest levels of RNA Polymerase II at human microRNA genes are within the ribosomal gene repeat arrays (rDNAs). Alignment of public nascent transcript data to the hs1 human genome assembly reveals a 50-nucleotide transcript for both miR-1275 and miR-6724, which exits from the nucleus with exceptional rapidity. We show that the miR-1275/miR-6724 transcription unit is closely flanked by CCCTC-binding factor (CTCF) within a <400-bp span of the rDNA spacer promoter. MiR-1275/miR-6724 and microRNA precursors expressed from the 5 External Transcribed Spacer (5ETS) are exported independently of known RNA processing activities and are detected in exosomes and as circulating cancer biomarkers. We propose that rDNA spacer promoter and 5ETS microRNA genes have evolved for general regulatory functions in recipient cells.
]]></description>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Henikoff, J. G.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.10.687711</dc:identifier>
<dc:title><![CDATA[Superabundant microRNAs are transcribed from human rDNA spacer promoters insulated by CTCF]]></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/10.1101/2025.11.10.687731v1?rss=1">
<title>
<![CDATA[
N6-methyladenosine regulation of mRNA translation is essential for early human erythropoiesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.10.687731v1?rss=1</link>
<description><![CDATA[
N6-methyladenosine (m6A) is an abundant modification of mRNA with important regulatory roles in normal and malignant hematopoiesis. We previously reported that in human erythroid leukemia (HEL) cells, m6A mRNA marking selectively regulates translation of essential erythropoiesis genes required for in vitro differentiation and human erythroid colony formation. Here, we further investigated the timing and nature of requirement for m6A-methyltransferase (MTase) activity during human erythropoiesis, using a standardized in vitro erythroid differentiation assay for hHSPCs. We identified two critical m6A regulated developmental windows in BFU-E and during the transition from CFU-E to proerythroblasts. These windows of m6A-MTase requirement coincide with rising global m6A levels, which peak in proerythroblasts. After proerythroblast formation, however, m6A -MTase activity is dispensable for differentiation, proliferation, and survival. In BFU-E, m6A-MTase promotes proliferation but is dispensable for differentiation, while, in CFU-E, both m6A -MTase and the YTHDF family of m6A readers are essential for differentiation to proerythroblasts. Mechanistically, in CFU-E, m6A MTase activity enhances translation of ribosomal and oxidative phosphorylation (OXPHOS) genes, thereby elevating global protein synthesis rates and enabling efficient erythroblast formation. We propose that this form of translational regulation by m6A emerged as an evolutionary adaptation to meet the high translational demands of human erythropoiesis.
]]></description>
<dc:creator>Kuppers, D.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Wladyka, C.</dc:creator>
<dc:creator>Ge, R.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Peng, Y.</dc:creator>
<dc:creator>Su, R.</dc:creator>
<dc:creator>Wilhite, A.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>He, C.</dc:creator>
<dc:creator>Hsieh, A.</dc:creator>
<dc:creator>Paddison, P.</dc:creator>
<dc:date>2025-11-12</dc:date>
<dc:identifier>doi:10.1101/2025.11.10.687731</dc:identifier>
<dc:title><![CDATA[N6-methyladenosine regulation of mRNA translation is essential for early human erythropoiesis]]></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/10.1101/2025.11.12.687706v1?rss=1">
<title>
<![CDATA[
Elucidating Neurodevelopmental Trajectories in Cancer with Topic Modeling: Revealing Persistent External Granule Layer Lineages in Medulloblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.12.687706v1?rss=1</link>
<description><![CDATA[
The cerebellar rhombic lip generates cerebellar progenitors and neurons that ultimately differentiate to comprise over half of all neurons in the adult human brain. Standard clustering approaches often fragment or miss rhombic lip progenitor populations entirely due to their transient nature, small size, and rapid state transitions, leaving fundamental questions unanswered about normal cerebellar development and how such processes may be hijacked in pediatric brain cancer. Medulloblastoma, the most common malignant pediatric brain tumor, affects approximately 500 children annually in the United States with overall survival rates varying dramatically by subgroup. Sonic hedgehog (SHH) medulloblastoma, comprising 25-30% of cases, arises from rhombic lip-derived granule neuron precursors (GNP) within the external granule layer (EGL) and has particularly poor outcomes in several subtypes (5-year survival [~]41%). Using our topic modeling framework on over one million fetal cerebellar nuclei, we identify proliferative rhombic lip and EGL states that bifurcate into distinct glial and neuronal lineages through intermediate progenitors and capture a portion of the developmental spectrum form outer EGL (oEGL) proliferation through inner EGL (iEGL) differentiation. These developmental signatures (topics) persist in medulloblastoma, validating GNP origins of SHH tumors and revealing age-specific molecular programs that correspond to distinct stages of EGL development within SHH subtypes. Our transferable framework enables systematic comparison of developmental and disease states across technologies without data integration, solving a fundamental challenge as genomic atlases expand.
]]></description>
<dc:creator>Rajendran, A.</dc:creator>
<dc:creator>Haldipur, P.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Grama, K.</dc:creator>
<dc:creator>Subramanian, S. S.</dc:creator>
<dc:creator>Galan, L. M.</dc:creator>
<dc:creator>Johnson, D.</dc:creator>
<dc:creator>Aldinger, K. A.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Millen, K. J.</dc:creator>
<dc:creator>Gennari, J. H.</dc:creator>
<dc:creator>Pattwell, S. S.</dc:creator>
<dc:date>2025-11-13</dc:date>
<dc:identifier>doi:10.1101/2025.11.12.687706</dc:identifier>
<dc:title><![CDATA[Elucidating Neurodevelopmental Trajectories in Cancer with Topic Modeling: Revealing Persistent External Granule Layer Lineages in Medulloblastoma]]></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/10.1101/2025.11.15.688648v1?rss=1">
<title>
<![CDATA[
E2F1-3 activate polyomavirus early transcription and viral replication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.15.688648v1?rss=1</link>
<description><![CDATA[
Merkel cell polyomavirus (MCPyV) is a DNA virus that establishes a persistent asymptomatic infection during childhood and can cause Merkel cell carcinoma (MCC) later in life. Its Large and Small Tumor antigens (LT, ST), splice variants of a common viral early transcript, drive viral replication and tumorigenesis by binding to and perturbing the function of host proteins. LT binds and inhibits RB1, deregulating E2F activity and host cell cycle control to permit viral replication during S phase. While the functions of LT and ST are relatively well characterized, how their expression is controlled is poorly defined. Here, we discovered that E2F1-3/DP1 dimers bind the MCPyV Non-Coding Control Region (NCCR) via a consensus E2 site close to the LT/ST transcriptional start site. Inhibiting E2F-NCCR binding, either by deleting the E2 site or treatment with a E2F small molecule inhibitor, downregulated LT/ST mRNA and protein expression in MCC cells and in 293A cells transfected with MCPyV. Our findings reveal an E2F/LT/RB1 positive feedback loop that appears to have evolved to support viral replication and is hijacked in MCC cells to promote cellular proliferation. Furthermore, we identified E2 sites in the NCCRs of PyVs closely related to MCPyV, including murine PyV, which mediate E2F/DP binding and potentiate viral early transcription. E2F/DP also bound weakly to the SV40 and BKPyV NCCRs, despite lacking an E2 site. Our findings challenge the prevailing model that PyV LT expression drives S phase entry and suggest, in contrast, that S phase entry stimulates PyV early transcription and replication.

Significance statementPolyomaviruses (PyVs) express Large and Small Tumor antigens (LT, ST), splice variants of a common viral early transcript, that drive viral replication and tumorigenesis by binding and perturbing the function of host cells. LTs bind and inhibit RB1 via conserved LxCxE motifs, deregulating E2F activity and host cell cycle control to permit viral replication. Here, we discovered that E2F1-3 bind to the Non-Coding Control Regions (NCCRs) of many, but not all, PyVs to activate LT/ST transcription, revealing an E2F/LT/RB1 positive feedback loop that appears to have evolved to promote viral replication. Our findings challenge the existing paradigm that PyV LT expression drives S phase entry and suggest, in contrast, that S phase entry stimulates PyV LT/ST transcription and replication.
]]></description>
<dc:creator>Salisbury, N. J. H.</dc:creator>
<dc:creator>Amonkar, S.</dc:creator>
<dc:creator>Roman, A.</dc:creator>
<dc:creator>Galloway, D. A.</dc:creator>
<dc:date>2025-11-16</dc:date>
<dc:identifier>doi:10.1101/2025.11.15.688648</dc:identifier>
<dc:title><![CDATA[E2F1-3 activate polyomavirus early transcription and viral replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.14.688517v1?rss=1">
<title>
<![CDATA[
Insights into the Datasets, Tools, and Training Needs of the AnVIL Community: 2024 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.14.688517v1?rss=1</link>
<description><![CDATA[
The NHGRI Genomic Data Science Analysis, Visualization, and Informatics Lab-space (AnVIL) provides a secure cloud-based environment where research and education communities can analyze genomic and biomedical data. The platform supports a wide range of data analysis as well as the ability to safely store and access data in compliance with NIH policies. Work on the AnVIL platform can be easily shared to promote reproducible science and collaboration. The purpose of this study is to better understand the current user base of the AnVIL platform. The AnVIL Community Poll aimed to collect baseline information, identify development opportunities, guide the prioritization of user support strategies, and succinctly but comprehensively describe the current AnVIL Community. The AnVIL Team disseminated the inaugural AnVIL Community Poll by sharing it broadly on social media and through AnVIL and related consortia mailing lists. We categorized respondents as either returning or potential users of the AnVIL platform (based on their provided usage description) and examined user experiences: specifically user backgrounds, technological comfort, research interests, computational needs, and preferences for training and support. Our sample of the AnVIL community found opportunities for platform adoption beyond the current user base and identified areas where training should be enhanced, training preferences, and user computational needs. Specifically, while most respondents were involved in human genomics research, there may be potential for growth in adoption of the platform by prioritizing materials to support clinical researchers. All respondents felt availability of specific tools or datasets was a key feature of the platform. The broader community may also benefit from further development or showcasing of resources to facilitate cost management, finding and incorporating analysis tools, and data import. Our sample greatly preferred virtual training opportunities and returning users of the platform foresaw needing large amounts of storage. This poll provided an insightful snapshot of the current state of the AnVIL and demonstrated areas where the AnVIL Team can take specific steps to address barriers related to platform adoption and further support the existing and varied AnVIL Community. This work can be built upon through user interviews, community discussion, and coordinating a recurring poll.
]]></description>
<dc:creator>Isaac, K. J.</dc:creator>
<dc:creator>Cox, K. E. L.</dc:creator>
<dc:creator>Ho, K. Y.</dc:creator>
<dc:creator>Humphries, E. M.</dc:creator>
<dc:creator>Kucher, N.</dc:creator>
<dc:creator>Leek, J. T.</dc:creator>
<dc:creator>Mosher, S.</dc:creator>
<dc:creator>Schatz, M. C.</dc:creator>
<dc:creator>Tan, F. J.</dc:creator>
<dc:creator>Hoffman, A. M.</dc:creator>
<dc:date>2025-11-17</dc:date>
<dc:identifier>doi:10.1101/2025.11.14.688517</dc:identifier>
<dc:title><![CDATA[Insights into the Datasets, Tools, and Training Needs of the AnVIL Community: 2024]]></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/10.1101/2025.11.19.689381v1?rss=1">
<title>
<![CDATA[
Targeting peptide-MHC complexes with designed T cell receptors and antibodies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.19.689381v1?rss=1</link>
<description><![CDATA[
Class I major histocompatibility complexes (MHCs), expressed on the surface of all nucleated cells, present peptides derived from intracellular proteins for surveillance by T cells. The precise recognition of foreign or mutated peptide-MHC (pMHC) complexes by T cell receptors (TCRs) is central to immune defense against pathogens and tumors. Although patient-derived TCRs specific for cancer-associated antigens have been used to engineer tumor-targeting therapies, their reactivity toward self- or near-self antigens may be constrained by negative selection in the thymus. Here, we introduce a structure-based deep learning framework, ADAPT (Antigen-receptor Design Against Peptide-MHC Targets), for the design of TCRs and antibodies that bind to pMHC targets of interest. We evaluate the ADAPT pipeline by designing and characterizing TCRs and antibodies against a diverse panel of pMHCs. Cryogenic electron microscopy structures of two designed antibodies bound to their respective pMHC targets demonstrate atomic-level accuracy at the recognition interface, supporting the robustness of our structure-based approach. Computationally designed TCRs and antibodies targeting pMHC complexes could enable a broad range of therapeutic applications, from cancer immunotherapy to autoimmune disease treatment, and insights gained from TCR-pMHC design should advance predictive understanding of TCR specificity with implications for basic immunology and clinical diagnostics.
]]></description>
<dc:creator>Motmaen, A.</dc:creator>
<dc:creator>Jude, K. M.</dc:creator>
<dc:creator>Wang, N.</dc:creator>
<dc:creator>Minervina, A.</dc:creator>
<dc:creator>Feldman, D.</dc:creator>
<dc:creator>Lichtenstein, M. A.</dc:creator>
<dc:creator>Ebenezer, A.</dc:creator>
<dc:creator>Correnti, C.</dc:creator>
<dc:creator>Thomas, P. G.</dc:creator>
<dc:creator>Garcia, K. C.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:date>2025-11-20</dc:date>
<dc:identifier>doi:10.1101/2025.11.19.689381</dc:identifier>
<dc:title><![CDATA[Targeting peptide-MHC complexes with designed T cell receptors and antibodies]]></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/10.1101/2025.11.20.688607v1?rss=1">
<title>
<![CDATA[
Orchestrating Spatial Transcriptomics Analysis with Bioconductor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.688607v1?rss=1</link>
<description><![CDATA[
Spatial transcriptomics technologies provide spatially-resolved measurements of gene expression through assays that can either target selected genes or capture transcriptome-wide expression profiles. The complexity and variability of these technologies and their associated data necessitate multi-step workflows integrating diverse computational methods and software packages. We provide a freely accessible, open-source, continuously updated and tested online book containing reproducible code examples, datasets, and discussion about data analysis workflows for spatial omics data using Bioconductor in R, including interoperability with Python.
]]></description>
<dc:creator>Crowell, H. L.</dc:creator>
<dc:creator>Dong, Y.</dc:creator>
<dc:creator>Billato, I.</dc:creator>
<dc:creator>Cai, P.</dc:creator>
<dc:creator>Emons, M.</dc:creator>
<dc:creator>Gunz, S.</dc:creator>
<dc:creator>Guo, B.</dc:creator>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Mahmoud, A.</dc:creator>
<dc:creator>Manukyan, A.</dc:creator>
<dc:creator>Pages, H.</dc:creator>
<dc:creator>Panwar, P.</dc:creator>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Sargeant, C. J.</dc:creator>
<dc:creator>Shepherd Kern, L.</dc:creator>
<dc:creator>Ramos, M.</dc:creator>
<dc:creator>Sun, J.</dc:creator>
<dc:creator>Totty, M.</dc:creator>
<dc:creator>Carey, V. J.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Collado-Torres, L.</dc:creator>
<dc:creator>Ghazanfar, S.</dc:creator>
<dc:creator>Hansen, K. D.</dc:creator>
<dc:creator>Martinowich, K.</dc:creator>
<dc:creator>Maynard, K. R.</dc:creator>
<dc:creator>Patrick, E.</dc:creator>
<dc:creator>Righelli, D.</dc:creator>
<dc:creator>Risso, D.</dc:creator>
<dc:creator>Tiberi, S.</dc:creator>
<dc:creator>Waldron, L.</dc:creator>
<dc:creator>Gottardo, R.</dc:creator>
<dc:creator>Robinson, M. D.</dc:creator>
<dc:creator>Hicks, S. C.</dc:creator>
<dc:creator>Weber, L. M.</dc:creator>
<dc:date>2025-11-21</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.688607</dc:identifier>
<dc:title><![CDATA[Orchestrating Spatial Transcriptomics Analysis with Bioconductor]]></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/10.1101/2025.11.20.689567v1?rss=1">
<title>
<![CDATA[
Tom70-mediated mitochondria-nuclear envelope contacts regulate nuclear pore complex inheritance during gametogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689567v1?rss=1</link>
<description><![CDATA[
Gametogenesis rejuvenates the cellular lineage and excludes senescence-associated factors from gametes. In Saccharomyces cerevisiae, this involves sequestration of nuclear constituents into the Gametogenesis-Uninherited Nuclear Compartment (GUNC), which is excluded from gametes. Here we identify the conserved mitochondrial import receptor Tom70 as a key regulator of GUNC-mediated exclusion. Loss of TOM70 disrupts the sequestration of nuclear pore complexes, but not senescence-associated aggregates and nucleolar components, into the GUNC. Tom70s role appears independent of its canonical function in mitochondrial import and instead reflects a meiosis-specific requirement for mitochondria-nuclear envelope tethering. During meiosis II, Tom70 concentrates around the GUNC, where it recruits the nuclear envelope tethering protein Cnm1. Loss of CNM1 partially phenocopies tom70{Delta}, consistent with parallel tethering interactions. These findings uncover a previously unrecognized organelle contact-dependent pathway that remodels the nuclear envelope to support selective nuclear inheritance. More broadly, they highlight how organelle contacts integrate with nuclear quality control to safeguard gamete integrity.
]]></description>
<dc:creator>Ruediger, C. T.</dc:creator>
<dc:creator>Styler, B. S.</dc:creator>
<dc:creator>Sawyer, E. S.</dc:creator>
<dc:creator>Spiri, S.</dc:creator>
<dc:creator>King, G. A.</dc:creator>
<dc:creator>Walsh, M. E.</dc:creator>
<dc:creator>Brar, G. A.</dc:creator>
<dc:creator>Jorgens, D. M.</dc:creator>
<dc:creator>Ünal, E.</dc:creator>
<dc:date>2025-11-21</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689567</dc:identifier>
<dc:title><![CDATA[Tom70-mediated mitochondria-nuclear envelope contacts regulate nuclear pore complex inheritance during gametogenesis]]></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/10.1101/2025.11.20.689586v1?rss=1">
<title>
<![CDATA[
3D pathology-guided microdissection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689586v1?rss=1</link>
<description><![CDATA[
Traditional micro- and macro-dissection techniques enable the extraction of localized regions in thin tissue sections for molecular analysis. Despite the growing use of 3D microscopy, analogous methods for volumetric microdissection are lacking. We have developed a 3D microdissection method based on computer numerical controlled (CNC) milling integrated with open-top light-sheet microscopy. We demonstrate the ability to study tumor evolution along convoluted 3D branching architectures, which is inaccessible to 2D methods.
]]></description>
<dc:creator>Hsieh, H.-C.</dc:creator>
<dc:creator>Gao, G.</dc:creator>
<dc:creator>Han, Q.</dc:creator>
<dc:creator>Brenes, D.</dc:creator>
<dc:creator>Baraznenok, E.</dc:creator>
<dc:creator>Yan, R.</dc:creator>
<dc:creator>Serafin, R.</dc:creator>
<dc:creator>Bishop, K. W.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Konnick, E. Q.</dc:creator>
<dc:creator>Pritchard, C. C.</dc:creator>
<dc:creator>Figiel, S.</dc:creator>
<dc:creator>Hamdy, F. C.</dc:creator>
<dc:creator>Mills, I. G.</dc:creator>
<dc:creator>Reder, N. P.</dc:creator>
<dc:creator>Reddi, D. M.</dc:creator>
<dc:creator>Paulson, T. G.</dc:creator>
<dc:creator>Grady, W. M.</dc:creator>
<dc:creator>Valk, J. E.</dc:creator>
<dc:creator>True, L. D.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Rao, S. R.</dc:creator>
<dc:creator>Woodcock, D. J.</dc:creator>
<dc:creator>Liu, J. T. C.</dc:creator>
<dc:date>2025-11-21</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689586</dc:identifier>
<dc:title><![CDATA[3D pathology-guided microdissection]]></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/10.1101/2025.11.21.689752v1?rss=1">
<title>
<![CDATA[
Contemporary HIV-1 envelope pseudovirus panels for detecting and assessing B cell lineages with broadly neutralizing antibody potential 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689752v1?rss=1</link>
<description><![CDATA[
Although a protective HIV-1 vaccine has not yet been realized, significant progress has been made in vaccine designs that trigger B cell lineages with potential to produce broadly neutralizing antibodies (bnAbs). Advancing these strategies by optimizing vaccine boosting regimens requires early detection of maturing antibodies with neutralizing activity against native envelope glycoprotein (Env) trimers and streamlined strategies to identify antibodies as they begin to manifest desired levels of breadth and potency. Thus, we designed three types of pseudovirus screening panels based on Envs of contemporary HIV-1 isolates to facilitate detection of bnAb lineages that are on favorable trajectories during a vaccination course. The panels were selected from Tier 2 Transmitted Founder Lineage (TFL) HIV-1 Envs from placebo participants in the Antibody Mediated Prevention (AMP) efficacy trials. Using 15 bnAbs to evaluate the neutralization sensitivity of the viruses, we selected 8-member bnAb class-specific panels most sensitive to bAbs representing their class: V2-apex, V3-glycan, CD4-receptor binding site (CD4bs), Membrane-Proximal External Region (MPER), or fusion peptide (FP). Next, we combined the most sensitive viruses among the class-specific panels to create a 12-virus panel to enable optimal detection of low-titer bnAb activity across epitope specificities. Finally, as HIV-1 continues to evolve greater levels of antigenic diversity and as current global pseudoviruses bnAb panels rely on viruses collected more than twenty years ago, we showed the importance of using contemporary viral panels to assess bnAb breadth and potency and designed a 12-virus panel representative of the spectrum neutralization profiles among AMP placebo viruses. We characterized pseudoviruses bearing each selected Env using standardized human sera to confirm their Tier 2 status and biological relevance. These updated panels enable sensitive screening of neutralization activity in vaccine studies and can also provide a realistic assessment of the expected breadth and potency of maturing responses against contemporary HIV-1 Envs.

AUTHOR SUMMARYA primary goal of HIV-1 vaccine research is to elicit neutralizing antibodies that can prevent infection, but HIV-1 is highly variable and so stimulating responses that can work effectively against the diverse array of HIV-1 variants remains an unsolved challenge. However, the HIV-1 vaccine field has made significant progress in terms of stimulating B cell lineages that produce antibodies known to have features able to ultimately generate potent broadly neutralizing antibodies. Inducing such precursor B cells is just the first step, as B cells need evolve in response to an antigenic stimulus through a process called affinity maturation to acquire potency and breadth against the natural diversity of circulating HIV-1 strains. The next step is to discover and optimize vaccine strategies that will selectively induce such maturation. To facilitate this work, we have designed panels of biologically relevant, contemporary HIV-1 Envelope pseudotyped viruses that will enable sensitive detection of neutralizing antibodies in human and animal vaccine studies. We also have defined a small screening panel intended to provide a quick but realistic assessment of the potential of a vaccine-stimulated antibody response or a monoclonal antibody being evaluated for clinical use to effectively counter and block currently circulating HIV-1 variants.
]]></description>
<dc:creator>Korber, B.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Mkhize, N. N.</dc:creator>
<dc:creator>Greene, K.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Shen, X.</dc:creator>
<dc:creator>Domin, E.</dc:creator>
<dc:creator>Tang, H.</dc:creator>
<dc:creator>Theiler, J.</dc:creator>
<dc:creator>Wagh, K.</dc:creator>
<dc:creator>Moore, P. L.</dc:creator>
<dc:creator>Williamson, C.</dc:creator>
<dc:creator>Mullins, J. I.</dc:creator>
<dc:creator>Doria-Rose, N. A.</dc:creator>
<dc:creator>Montefiori, D.</dc:creator>
<dc:creator>Giorgi, E. E.</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689752</dc:identifier>
<dc:title><![CDATA[Contemporary HIV-1 envelope pseudovirus panels for detecting and assessing B cell lineages with broadly neutralizing antibody potential]]></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/10.1101/2025.11.25.689574v1?rss=1">
<title>
<![CDATA[
The BRCA1- RAD51 Axis Regulates SCAI/REV3 Dependent Replication Fork Maintenance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.25.689574v1?rss=1</link>
<description><![CDATA[
Replication stress is a key contributor to genomic instability and cancers. BRCA1 has well established roles in protecting stalled forks against degradation. Here we show that BRCA1 has a fork protection-independent role via regulation of RAD51 in a manner that confers dependency on SCAI and REV3. SCAI loss leads to elevated DNA breaks, genomic instability and cell death upon DNA damage. We show that the increased DNA breaks occur in S phase, consistent with fork breakage. BRCA1 drives SLX4-SLX1-ERCC1 mediated DNA break formation in the absence of SCAI and REV3, which are required to maintain integrity of stalled forks for efficient restart. Domain analysis revealed that the increased break signaling seen in the absence of SCAI does not require binding to resection factors like CtIP. Surprisingly, loss of fork reversal factors leads to additive increases in damage signaling and elevated fork shortening in the absence of SCAI/REV3 in a manner that depends on RAD51 activity. We propose that Protexin may be required to replicate through and maintain stalled replication forks at fragile genomic regions. Failure to do so leads to increased DNA breakage and genomic instability.
]]></description>
<dc:creator>Unterseher, C.</dc:creator>
<dc:creator>Tsuchida, H.</dc:creator>
<dc:creator>Bosire, R.</dc:creator>
<dc:creator>Jin, X.</dc:creator>
<dc:creator>Kieffer, S.</dc:creator>
<dc:creator>Adeyemi, R.</dc:creator>
<dc:date>2025-11-27</dc:date>
<dc:identifier>doi:10.1101/2025.11.25.689574</dc:identifier>
<dc:title><![CDATA[The BRCA1- RAD51 Axis Regulates SCAI/REV3 Dependent Replication Fork Maintenance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.29.690828v1?rss=1">
<title>
<![CDATA[
The type VI secretion system governs strain maintenance in a wild mammalian gut microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.29.690828v1?rss=1</link>
<description><![CDATA[
Bacteria inhabiting the mammalian gut coexist in dense communities where contact-dependent antagonism mechanisms are widespread. The type VI secretion system (T6SS) is an interbacterial toxin delivery pathway prevalent among gut Bacteroidales, yet its function in naturally evolved microbiomes remains poorly defined. Here, we examine the physiological role of the T6SS in Bacteroides within a gut community derived from wild mice (the WildR microbiome). Using newly developed genetic tools and a strategy for functional replacement of strains within the WildR community, we demonstrate that the WildR isolate B. acidifaciens employs a T6SS to antagonize co-resident Bacteroidales. We also show that loss of T6SS function compromises the long-term maintenance of B. acidifaciens in the community but not its initial colonization, establishing the system as a determinant of strain persistence. The T6SS we identified resides on an integrative and conjugative element (ICE). ICE-seq, a targeted sequencing approach, reveals that the T6SS-ICE is distributed among select Bacteroidales and Muribaculaceae species in the WildR microbiome, between which it appears to be recently exchanged. We also show that transfer of the T6SS-ICE to WildR isolate Phocaeicola vulgatus confers transient colonization benefits in mice, but an eventual fitness cost. Our findings demonstrate that the T6SS can stabilize the presence of specific strains within a complex, co-evolved gut microbiome, yet its value is context dependent and constrained by the ecological and physiological landscape of the host community.
]]></description>
<dc:creator>Shen, B. A.</dc:creator>
<dc:creator>Asfahl, K. L.</dc:creator>
<dc:creator>Lim, B.</dc:creator>
<dc:creator>Bertolli, S. K.</dc:creator>
<dc:creator>Minot, S. S.</dc:creator>
<dc:creator>Radey, M. C.</dc:creator>
<dc:creator>Penewit, K.</dc:creator>
<dc:creator>Ngo, B.</dc:creator>
<dc:creator>Salipante, S. J.</dc:creator>
<dc:creator>Johnston, C. D.</dc:creator>
<dc:creator>Peterson, S. B.</dc:creator>
<dc:creator>Goodman, A. D.</dc:creator>
<dc:creator>Mougous, J. D.</dc:creator>
<dc:date>2025-11-30</dc:date>
<dc:identifier>doi:10.1101/2025.11.29.690828</dc:identifier>
<dc:title><![CDATA[The type VI secretion system governs strain maintenance in a wild mammalian gut microbiome]]></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/10.64898/2025.12.02.691676v1?rss=1">
<title>
<![CDATA[
Dissecting type I and II interferon impacts on human immune cells in disease by a cell type-specific interferon response atlas 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.02.691676v1?rss=1</link>
<description><![CDATA[
Interferons (IFNs) orchestrate diverse immune responses, but distinguishing individual IFN contributions in human transcriptomic data is challenging due to overlapping interferon-stimulated gene (ISG) signatures and limited cell-type-specific datasets. To address this, we generated a single-cell transcriptomic atlas of IFN responses by stimulating primary human T, B, NK, and CD14 monocytes with IFN-I, IFN-II, and IFN-III. This revealed core and cell-type-specific ISG programs across 13 subsets, highlighting distinct functions of IFNs. We developed an algorithm to separate IFN-I and IFN-II activity in transcriptomic data. Applied to multiple myeloma samples, it showed elevated IFN-I and IFN-II responses, with induction therapy reducing only IFN-I. Extending to multiple disease datasets provided a cross-disease overview of IFN-I and IFN-II activities and revealed increased IFN-II activities in T cells during lupus flares. This resource and the accompanying analytical framework enable dissection of IFN-driven transcriptional programs in a cell-type specific manner in human disease.
]]></description>
<dc:creator>Moss, N.</dc:creator>
<dc:creator>Sakai, C.</dc:creator>
<dc:creator>Kaul, S. N.</dc:creator>
<dc:creator>Graybuck, L. T.</dc:creator>
<dc:creator>Rachid Zaim, S.</dc:creator>
<dc:creator>Angus-Hill, M. L.</dc:creator>
<dc:creator>He, Y. D.</dc:creator>
<dc:creator>Layton, E. D.</dc:creator>
<dc:creator>Bouvatte, P.</dc:creator>
<dc:creator>Wittig, P. J.</dc:creator>
<dc:creator>La France, C. M.</dc:creator>
<dc:creator>Peng, T.</dc:creator>
<dc:creator>Glass, M. C.</dc:creator>
<dc:creator>Krishnan, U.</dc:creator>
<dc:creator>Chander, A.</dc:creator>
<dc:creator>Kawelo, E. K.</dc:creator>
<dc:creator>Garber, J.</dc:creator>
<dc:creator>Reading, J.</dc:creator>
<dc:creator>Anover-Sombke, S. D.</dc:creator>
<dc:creator>Kwok, M.</dc:creator>
<dc:creator>Green, D. J.</dc:creator>
<dc:creator>Goldrath, A. W.</dc:creator>
<dc:creator>Sigvardsson, M.</dc:creator>
<dc:creator>Skene, P. J.</dc:creator>
<dc:creator>Li, X.-j.</dc:creator>
<dc:creator>Torgerson, T. R.</dc:creator>
<dc:creator>Kuan, E. L.</dc:creator>
<dc:date>2025-12-05</dc:date>
<dc:identifier>doi:10.64898/2025.12.02.691676</dc:identifier>
<dc:title><![CDATA[Dissecting type I and II interferon impacts on human immune cells in disease by a cell type-specific interferon response atlas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-05</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/10.64898/2025.12.08.693072v1?rss=1">
<title>
<![CDATA[
Armoring STEAP1 CAR T cells with IL-18 potentiates antitumor activity in Ewing sarcoma 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.08.693072v1?rss=1</link>
<description><![CDATA[
BackgroundEwing sarcoma (EwS) is a highly aggressive cancer driven by the EWS::FLI1 fusion oncoprotein affecting children, adolescents, and young adults. Six transmembrane epithelial antigen 1 (STEAP1) is a cell surface antigen transcriptionally controlled by EWS::FLI1 that is broadly expressed in EwS, positioning it as a rational immunotherapy target. However, translating CAR T therapy to solid tumors requires overcoming barriers to potency while maintaining safety.

MethodsAnalyses of transcriptome and proteome data were performed to evaluate the effects of EWS::FLI1 perturbation on STEAP1 expression at the transcript and protein levels in EwS models. STEAP1 expression was validated in EwS patient tissues by immunohistochemistry. Second-generation STEAP1-BB{zeta} CAR T cells were tested in orthotopic and disseminated EwS xenograft models. To enhance antitumor activity, an IL-18-armored STEAP1 CAR was engineered. Dose-dependent therapeutic efficacy and safety were evaluated through measurement of tumor burden, survival, and observation for gross toxicities.

ResultsSTEAP1 was expressed in [~]97% of primary EwS tumors and directly associated with EWS::FLI1 fusion protein expression in EwS cell lines. In orthotopic EwS models, STEAP1 CAR T cells induced complete tumor regression at 5 x 106 cells. In disseminated disease models, responses were dose-dependent with no evidence of antigen loss. Notably, IL-18 armored STEAP1 CAR T cells achieved complete responses in [~]80% of mice at a reduced dose of 106 cells without overt toxicity.

ConclusionsThese data establish STEAP1 as a clinically relevant and highly expressed target in EwS and demonstrate that IL-18 armoring significantly improves CAR T cell efficacy by enhancing potency evident through antitumor activity at reduced cell dose. STEAP1 CAR T cells are currently under evaluation in a first-in-human phase 1/2 dose-escalation clinical trial for metastatic castration-resistant prostate cancer (NCT06236139) and these studies support future clinical translation of STEAP1 CAR T cell therapy for relapsed/refractory EwS.
]]></description>
<dc:creator>Bhatia, V.</dc:creator>
<dc:creator>Tsao, A.</dc:creator>
<dc:creator>Chong, T.</dc:creator>
<dc:creator>Challita, P. P.</dc:creator>
<dc:creator>Liang, K.</dc:creator>
<dc:creator>Sayar, E.</dc:creator>
<dc:creator>Huang, J.</dc:creator>
<dc:creator>Lawlor, E. R.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Nabet, B.</dc:creator>
<dc:creator>Lee, J. K.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.08.693072</dc:identifier>
<dc:title><![CDATA[Armoring STEAP1 CAR T cells with IL-18 potentiates antitumor activity in Ewing sarcoma]]></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/10.64898/2025.12.12.691241v1?rss=1">
<title>
<![CDATA[
Using experimental results of protein design to guide biomolecular energy-function development 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.12.691241v1?rss=1</link>
<description><![CDATA[
Computational models of macromolecules have many applications in biochemistry, but physical inaccuracies limit their utility. One class of models uses energy functions rooted in classical mechanics. The standard datasets used to train these models are limited in diversity, pointing to a need for new training data. Here, we sought to explore a new paradigm for training an energy function, where the Rosetta energy function was used to design de novo proteins. Experimental results on these designs were then used to identify failure modes of design, which were subsequently used as a "guiding principle" to retrain the energy function. Specifically, we examined a diverse set of de novo protein designs experimentally tested for their ability to stably fold, identifying unstable designs that were predicted to be stable by the Rosetta energy function. Using deep mutational scanning, we identified single amino-acid mutations that rescued the stability of these designs, providing insight into common failure modes of the energy function. We identified one key failure mode, involving steric clashing in protein cores. We identified similar overpacking when using Rosetta to refine high-resolution protein crystal structures, quantified the degree of overpacking, and refit a small set of energy-function parameters to better recapitulate native-like packing. Following fitting, we largely eliminated the failure mode in the refinement task, while retaining performance on other benchmarks, resulting in an updated version of the Rosetta energy function. This work shows how learning from protein designs can guide energy-function development.

Author summaryComputational models of macromolecules have many applications, such as predicting structures, predicting mutational effects, or designing new proteins. One type of model uses an energy function to explicitly model the physical forces at play. These models are trained using experimental data. However, available training data are limited in number and diversity, prompting a need for new sources of training data. In this paper, we explore a new paradigm for training an energy function, which involves using the energy function to design de novo proteins and then learning from which designs succeed and which fail when experimentally tested in the lab. We used experimental data to learn common failure modes in design, identified examples of a failure mode in a high-quality benchmark, and then used this benchmark to retrain the energy function. Using this strategy, we identified and largely resolved a bias in the Rosetta energy function that involved energetically unfavorable steric clashes in protein cores. Overall, this work helps establish a framework for how learning from design can be used to guide the development of macromolecular models.
]]></description>
<dc:creator>Haddox, H. K.</dc:creator>
<dc:creator>Rocklin, G. J.</dc:creator>
<dc:creator>Motta, F. C.</dc:creator>
<dc:creator>Strickland, D.</dc:creator>
<dc:creator>Halabiya, S. F.</dc:creator>
<dc:creator>Cordray, C.</dc:creator>
<dc:creator>Park, H.</dc:creator>
<dc:creator>Klavins, E.</dc:creator>
<dc:creator>Baker, D.</dc:creator>
<dc:creator>DiMaio, F.</dc:creator>
<dc:date>2025-12-13</dc:date>
<dc:identifier>doi:10.64898/2025.12.12.691241</dc:identifier>
<dc:title><![CDATA[Using experimental results of protein design to guide biomolecular energy-function development]]></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/10.64898/2025.12.12.693809v1?rss=1">
<title>
<![CDATA[
Dynamic evolution of EZHIP, an inhibitor of the Polycomb Repressive Complex 2 in mammals 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.12.693809v1?rss=1</link>
<description><![CDATA[
The Polycomb Repressive Complex 2 (PRC2) is an ancient, conserved chromatin-interacting complex that controls gene expression, facilitating differentiation and cellular identity during development. Its regulation is critical in most eukaryotes. EZHIP was recently characterized as a PRC2 inhibitor and  oncohistone mimic in mammals. Although Ezhip expression is typically restricted to the germline, its aberrant expression in pediatric brain tumors inhibits PRC2-mediated H3K27 methylation and drives disease progression. To gain a deeper understanding of its normal functions, we systematically examined Ezhip evolution across 70 mammals using comparative genomics, synteny analysis, and motif discovery. Bolstering previous work, we find that Ezhip originated and has been strictly retained on the X chromosome in placental mammals. In addition to the highly conserved H3K27M-like histone mimic motif, our motif analysis reveals seven previously unidentified EZHIP motifs, including a putative nuclear localization signal, and tandem repeats that are largely well-conserved in placental mammals, except in some lineages. We hypothesize that these motifs are also critical to EZHIPs functions, including in PRC2 interaction and inhibition. We show that Ezhip has evolved under strong diversifying selection in primates and underwent dynamic expansions and losses across species. Some paralogs, such as Ezhip2 in primates, also evolved under positive selection. Based on its evolutionary attributes and germ-cell expression, we propose that Ezhip arose and evolved rapidly due to inter-parental conflict over fetal development in utero in placental mammals. Our work provides a foundation for further investigations into EZHIPs essential, potentially multifaceted roles in mammalian reproduction and disease.
]]></description>
<dc:creator>Raman, P.</dc:creator>
<dc:creator>Khan, H.</dc:creator>
<dc:creator>Young, J. M.</dc:creator>
<dc:creator>Tsukiyama, T.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2025-12-13</dc:date>
<dc:identifier>doi:10.64898/2025.12.12.693809</dc:identifier>
<dc:title><![CDATA[Dynamic evolution of EZHIP, an inhibitor of the Polycomb Repressive Complex 2 in mammals]]></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/10.64898/2025.12.11.693522v1?rss=1">
<title>
<![CDATA[
Retention of a single Cenp-C gene in different syntenic locations in the montium group of Drosophila species 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.11.693522v1?rss=1</link>
<description><![CDATA[
Chromosome segregation in eukaryotes requires the orchestrated interaction of chromosomes with microtubules, mediated by the kinetochore multiprotein complex that assembles on specific chromosomal regions known as centromeres. In most eukaryotes, two centromeric proteins, CenH3 and Cenp-C, are essential for centromere function. In Drosophila, the localization of CenH3 (referred to as Cid in Drosophila) depends on its chaperone CAL1 and Cenp-C. Previous studies have shown that both Cid and Cenp-C underwent a coincident gene duplication and likely functional specialization in the Drosophila subgenus. Independently, Cid duplications led to Cid1, Cid3, and Cid4 paralogs in the montium group (Sophophora subgenus), but it is unknown whether this group also underwent parallel duplications of Cenp-C. Here, we investigate this possibility by analyzing sequenced genomes of 23 montium group species. We identified Cenp-C genes in five distinct syntenic loci; we named these genes Cenp-C1b, Cenp-C1c, Cenp-C1d, Cenp-C1e and Cenp-C3. Despite their distinct synteny, most montium group species only encode a single Cenp-C; their phylogeny mirrors the species phylogeny, and they appear to have retained Cenp-C protein motifs indicative of function. A closer examination revealed that these Cenp-C genes resulted from gene translocations or alternate retention (duplication followed by loss of the ancestral copy); only one species, D. vulcana, retains two intact Cenp-C paralogs. Therefore, unlike the Drosophila genus, the co-retention of three Cid paralogs in the montium group has not resulted in a coincident Cenp-C paralog co-retention. Our work highlights differences in functional retention and likely specialization of the two most conserved centromeric proteins in eukaryotes.
]]></description>
<dc:creator>Soares, R. F.</dc:creator>
<dc:creator>Chang, C.-H.</dc:creator>
<dc:creator>Koerich, L. B.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:creator>KUHN, G. C. S.</dc:creator>
<dc:date>2025-12-15</dc:date>
<dc:identifier>doi:10.64898/2025.12.11.693522</dc:identifier>
<dc:title><![CDATA[Retention of a single Cenp-C gene in different syntenic locations in the montium group of Drosophila species]]></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/10.64898/2025.12.11.693768v1?rss=1">
<title>
<![CDATA[
NKTR-255, a polymer-conjugated IL-15, synergizes with CAR-T cell therapy to activate endogenous anti-tumor immunity and improve tumor control 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.11.693768v1?rss=1</link>
<description><![CDATA[
CAR-T cells have yet to show widespread efficacy in solid tumors due in part to their poor persistence and loss of function in the tumor microenvironment. Further, heterogenous expression of most CAR target antigens in solid tumors can lead to escape of antigen-null tumors that resist CAR-T killing. Strategies to cooperatively boost both CAR-T and endogenous anti-tumor immunity could curb tumor escape and may be critical for achieving durable efficacy in cancer patients. NKTR-255 is a polymer-conjugated IL-15 with extended half-life that can boost endogenous T and NK cells, as well as CD19 CAR-T activity in B cell malignancies. However, whether NKTR-255 is sufficient to overcome CAR-T dysfunction in the suppressive solid tumor microenvironment, and how NKTR-255 and CAR-Ts together re-shape endogenous anti-tumor immunity, is not known. Using an autochthonous mouse model of ROR1+ lung adenocarcinoma, we show that NKTR-255 significantly boosted accumulation, reduced exhaustion, and improved function of tumor-infiltrating CAR-T cells. Compared with NKTR-255 or CAR-T treatment alone, combination of NKTR-255 and CAR-T therapy synergistically increased tumor-infiltrating CD11b+ cytotoxic NK cells, activated dendritic cells, and endogenous tumor-specific T cells that preserved a PD-1+Tcf1+ stem-like phenotype. Consequently, NKTR-255 and CAR-T combination therapy induced complete elimination of ROR1+ tumor and significantly improved survival, with enhanced tumor control dependent on activity of both CAR-Ts and endogenous T cells. Altogether, our data suggest that combining NKTR-255 with CAR-T therapy is a promising strategy to enhance both CAR-T and endogenous anti-tumor immunity to promote coordinated control of aggressive tumors.
]]></description>
<dc:creator>Nutt, W. S.</dc:creator>
<dc:creator>Kluesner, M. G.</dc:creator>
<dc:creator>Bingham, E.</dc:creator>
<dc:creator>Gad, E.</dc:creator>
<dc:creator>Miller, D.</dc:creator>
<dc:creator>Zepeda, V.</dc:creator>
<dc:creator>Snyder, A. J.</dc:creator>
<dc:creator>Marsh, S. A.</dc:creator>
<dc:creator>Liudahl, S. M.</dc:creator>
<dc:creator>Hoffman, M.</dc:creator>
<dc:creator>LeBlanc, L.</dc:creator>
<dc:creator>Volfbeyn, M. E.</dc:creator>
<dc:creator>Garrison, S. M.</dc:creator>
<dc:creator>Sarvothama, M.</dc:creator>
<dc:creator>Barry, K. C.</dc:creator>
<dc:creator>Headley, M. B.</dc:creator>
<dc:creator>Marcondes, M.</dc:creator>
<dc:creator>Srivastava, S.</dc:creator>
<dc:date>2025-12-15</dc:date>
<dc:identifier>doi:10.64898/2025.12.11.693768</dc:identifier>
<dc:title><![CDATA[NKTR-255, a polymer-conjugated IL-15, synergizes with CAR-T cell therapy to activate endogenous anti-tumor immunity and improve tumor control]]></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/10.64898/2025.12.16.694801v1?rss=1">
<title>
<![CDATA[
Metaplasia Enables Stomach Colonization by Fusobacterium animalis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.16.694801v1?rss=1</link>
<description><![CDATA[
Infection with Helicobacter pylori is the major risk factor for gastric cancer worldwide; yet the exact mechanisms behind this link remain unclear. H. pylori-associated tissue changes often disrupt the gastric microbiome, enabling secondary gastric colonization by oral bacteria. Among these secondary colonizers, Fusobacterium species have documented associations with several gastrointestinal cancers. We found that both F. animalis and F. nucleatum invade cultured human gastric adenocarcinoma cells, but F. animalis exhibited higher adherence and invasion, and hypoxic conditions promoted higher bacterial survival. Both adherence and invasion were inhibited by exogenous GalNAc, a glycan commonly observed in membrane glycoproteins of adenocarcinoma cells, and a target of the fusobacterial adhesin Fap2. Using a mouse model of gastric metaplasia, we found that F. animalis colonized gastric tissue only after metaplasia onset, growing in multispecies biofilms in the mucus layer, while F. nucleatum colonized neither healthy nor metaplastic gastric tissue. Metaplasia led to upregulation of Gal-GalNAc in the stomach, and reduced gastric acidity allowed higher F. animalis loads in this model. By contrast, inflammation and the presence of H. pylori did not significantly influence stomach colonization by F. animalis. Overall, our data support a model in which H. pylori-induced metaplasia makes the stomach susceptible to secondary infection by another cancer-associated microbe, F. animalis.
]]></description>
<dc:creator>Gomez-Garzon, C.</dc:creator>
<dc:creator>Chen, Q.</dc:creator>
<dc:creator>O'Brien, V. P.</dc:creator>
<dc:creator>Salama, N.</dc:creator>
<dc:date>2025-12-18</dc:date>
<dc:identifier>doi:10.64898/2025.12.16.694801</dc:identifier>
<dc:title><![CDATA[Metaplasia Enables Stomach Colonization by Fusobacterium animalis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.21.695820v1?rss=1">
<title>
<![CDATA[
PTEN deficiency linked to chromosome 10q loss leads to aggressive NF2mutant meningioma biology 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.21.695820v1?rss=1</link>
<description><![CDATA[
While most meningiomas are benign and can be surgically removed, a subset behaves aggressively, recurs quickly, and can ultimately be fatal. Recent work has focused on defining this aggressive group. To better characterize this clinically distinct, high-risk group, we analyzed bulk RNA sequencing (RNA-seq) from human meningiomas using a dimension-reduced reference landscape. We identified an NF2 mutant subtype enriched for chromosome 10q loss and low PTEN expression, both of which were strongly associated with shorter time to recurrence. To directly test causality, we engineered several genetically accurate, immunocompetent in vitro and in vivo mouse models with Pten knockdown or knockout. Across models, Pten loss markedly accelerated YAP1-driven tumorigenesis. Projection of Pten-deficient mouse tumors onto bulk and single-cell RNA-seq datasets from human meningiomas revealed that mouse tumors faithfully recapitulate the transcriptional programs of aggressive NF2 mutant meningiomas and robustly predicted clinical outcomes. Collectively, these findings demonstrate that PTEN loss cooperates with YAP1 activation to drive aggressive NF2 mutant meningioma biology, and that the high fidelity of this model to human disease establishes it as a robust platform for mechanistic investigation and therapeutic testing. More broadly, this work highlights the power of cross-species integration in both validating preclinical models and enhancing the translational relevance and utility of dimension-reduced reference landscapes across diverse disease types.

One Sentence SummaryPTEN loss causally drives aggressive NF2 mutant meningiomas, and cross-species integration provides a novel, high-fidelity system to validate preclinical models to human disease.
]]></description>
<dc:creator>Parrish, A. G.</dc:creator>
<dc:creator>Thirmanne, H. N.</dc:creator>
<dc:creator>Arora, S.</dc:creator>
<dc:creator>Herold-Mende, C.</dc:creator>
<dc:creator>Sievers, P.</dc:creator>
<dc:creator>Sahm, F.</dc:creator>
<dc:creator>Szulzewsky, F.</dc:creator>
<dc:creator>Holland, E.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.21.695820</dc:identifier>
<dc:title><![CDATA[PTEN deficiency linked to chromosome 10q loss leads to aggressive NF2mutant meningioma biology]]></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/10.64898/2025.12.21.695848v1?rss=1">
<title>
<![CDATA[
Mitochondrial ROS-induced metabolic alterations differentially regulate ferroptosis sensitivity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.21.695848v1?rss=1</link>
<description><![CDATA[
Ferroptosis is an iron-catalyzed lipid peroxidation (LP)-dependent cell death. Induction of mitochondrial ROS (mtROS) is crucial in the execution of ferroptosis, but the underlying mechanism remains unclear. Through utilizing the hepatocyte model and RNA-seq analysis, we determined mtROS-dependent metabolic changes that modulate ferroptosis sensitivity. Elevated mtROS production and LP suppressed glycolysis, fatty acid oxidation, and citric acid cycle activity, representing adaptive responses that protect cells from ferroptosis. On the other hand, mtROS-driven signaling impaired glutathione biosynthesis and downregulated genes involved in coenzyme Q10 (CoQ) biosynthesis, including those in the mevalonate pathway and CoQ8A, a key stabilizer of the CoQ biosynthetic complex. Importantly, silencing CoQ8A expression enhanced, whereas overexpression of CoQ8A reduced, ferroptosis susceptibility of hepatocytes and various cancer cell types. The mtROS-mediated downregulation of CoQ8A was dependent on farnesoid X receptor (FXR) and retinoid X receptors (RXRs). Collectively, our findings highlight that mtROS promotes ferroptosis, at least in part, by suppressing glutathione and CoQ biosynthesis.
]]></description>
<dc:creator>Banerjee, S.</dc:creator>
<dc:creator>Smith, M. R.</dc:creator>
<dc:creator>Wang, Y. I.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Gratz, D.</dc:creator>
<dc:creator>Tharp, G. K.</dc:creator>
<dc:creator>Kang, S.</dc:creator>
<dc:creator>He, P.</dc:creator>
<dc:date>2025-12-23</dc:date>
<dc:identifier>doi:10.64898/2025.12.21.695848</dc:identifier>
<dc:title><![CDATA[Mitochondrial ROS-induced metabolic alterations differentially regulate ferroptosis sensitivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.24.696395v1?rss=1">
<title>
<![CDATA[
Chromatin profiling for everyone: FFPE-CUTAC for the theory and practice of modern molecular biology 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.24.696395v1?rss=1</link>
<description><![CDATA[
In 2025, together with the Fred Hutch Summer Undergraduate Research and Summer High School Internship programs, we developed and implemented a laboratory genomics research experience to introduce students to modern molecular biology techniques and bioinformatics. The course centered around using a new method we had developed in 2023 that uses readily available fixed tissue sections on glass slides. Students performed a series of steps to tagment genomic locations of RNA Polymerase II and then used PCR to enrich libraries for next-generation sequencing in a core facility. Students then visualized their data in genomic browser tracks and assessed the results. At the end of the summer, students prepared and presented their work and experiences in seminar format to their cohorts. Overall, the technical simplicity of on-slide chromatin profiling introduced the students to laboratory practice and current techniques in genomics, bioinformatics, and medical sciences.
]]></description>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Henikoff, S.</dc:creator>
<dc:creator>Ahmad, K.</dc:creator>
<dc:date>2025-12-26</dc:date>
<dc:identifier>doi:10.64898/2025.12.24.696395</dc:identifier>
<dc:title><![CDATA[Chromatin profiling for everyone: FFPE-CUTAC for the theory and practice of modern molecular biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.29.692457v1?rss=1">
<title>
<![CDATA[
Generative AI Enables Breast Cancer Genomic Subtype Prediction from Histology Images 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.29.692457v1?rss=1</link>
<description><![CDATA[
Breast cancer subtyping is essential for precision oncology, influencing prognosis, treatment selection, and clinical trial design. The Integrative Subtype Classification (IC) categorizes breast tumors into groups with distinct long-term outcomes based on genomic and correlated transcriptomic features. This method relies on sequencing data, which, despite decreasing costs, is not always available in research or clinical settings. Here we introduce PATH-IC, a computational pathology model that predicts ER+ breast cancer IC subtype risk of relapse categories from routine histology data. We enhance the current state-of-the-art computational pathology approach with BERGERON, which leverages generative AI to correct class imbalance and reduce overfitting, showing that synthetic data improves PATH-ICs performance by the equivalent of 41% more real training samples. PATH-IC achieves a testing AUROC of 0.814, with predictions correlating to Oncotype DX scores and long-term relapse risk. Using attention-based model interpretation and CRAWFORD, a novel embedding-to-image foundation model, we demonstrate that PATH-IC identifies expected tumor microenvironment patterns for IC subtypes and highlights heterochromatin condensation as a key feature of high-risk tumors. Matched single-cell spatial transcriptomics confirm IC subtype-specific gene expression patterns identified by PATH-IC, including active metabolic, proliferative, and proteostasis pathways in the high-risk group. PATH-IC advances computational pathology through generative AI, enabling subtype inference from histopathology data.
]]></description>
<dc:creator>Simon, B. G.</dc:creator>
<dc:creator>Weiss, C. L. G.</dc:creator>
<dc:creator>Chan, D.</dc:creator>
<dc:creator>Mangiante, L.</dc:creator>
<dc:creator>Ma, Z.</dc:creator>
<dc:creator>Smith, N. H.</dc:creator>
<dc:creator>Meisner, A.</dc:creator>
<dc:creator>Rae, J. M.</dc:creator>
<dc:creator>Speers, C. W.</dc:creator>
<dc:creator>Albain, K. S.</dc:creator>
<dc:creator>Karakas, C.</dc:creator>
<dc:creator>Bean, G. R.</dc:creator>
<dc:creator>Mouron, S.</dc:creator>
<dc:creator>Quintela-Fandino, M.</dc:creator>
<dc:creator>Curtis, C.</dc:creator>
<dc:date>2025-12-29</dc:date>
<dc:identifier>doi:10.64898/2025.12.29.692457</dc:identifier>
<dc:title><![CDATA[Generative AI Enables Breast Cancer Genomic Subtype Prediction from Histology Images]]></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/10.64898/2025.12.29.696811v1?rss=1">
<title>
<![CDATA[
Pre-Human Immunodeficiency Virus (HIV) infection Th17 CD4+ T cells as predictors of early HIV disease progression 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.29.696811v1?rss=1</link>
<description><![CDATA[
Interleukin-17-producing T helper (Th17) CD4+ T cells are highly susceptible to HIV infection and are depleted early in people living with HIV. Here, we investigated whether systemic Th17 cell levels prior to HIV infection are associated with subsequent HIV disease progression. We analyzed archived cryopreserved peripheral blood mononuclear cells (PBMCs) collected within one year prior to HIV acquisition from participants enrolled in a South African cohort (HIV Vaccine Trials Network [HVTN] 503; n = 35) and an East African cohort (Partners Pre-exposure Prophylaxis/Couples Observational Study [PP/COS]; n = 32). Th17 cell frequencies were quantified by flow cytometry. In HVTN 503, higher pre-HIV IL-17+ CD4+ T cell frequencies were inversely correlated with CD4/CD8 ratio measured both within 180 days (Spearman rank R = -0.42, p = 0.012) and beyond 180 days (R = -0.55, p = 0.001) after HIV infection, and were associated with faster CD4+ T cells decline (adjusted hazard ratio [aHR] = 3.5, 95% CI: 1.2-9.9, p = 0.020). In contrast, no significant association with CD4 decline was observed in the PP/COS cohort (HR = 1.2, 95% CI: 0.4-3.4, p = 0.795). Sex-stratified analyses in HVTN 503 indicated a more pronounced association between pre-HIV IL-17+ CD4+ T cells and faster CD4 decline in males than females. In analyses combining all cohorts, higher pre-HIV IL-17+ CD4+ T cell frequencies remained associated with faster CD4 decline, particularly among younger participants (HR = 3.5; 95% CI: 1.35-9.22, p = 0.010). Pre-HIV IL-17+ CD4+ T cell frequencies were not associated with peak or set-point viral load in either cohort. Together, these findings suggest that pre-HIV Th17 cells abundance may influence subsequent HIV disease progression independently of early viral replication.

Author SummaryHIV infection leads to progressive damage of the immune system, but the rate at which this damage occurs varies widely between individuals. Understanding the factors that influence HIV disease progression is essential for improving prevention and treatment strategies. Previous studies have suggested that immune characteristics present before infection may shape disease outcomes after HIV acquisition. In this study, we examined whether the abundance of IL-17-producing CD4+ T cells, known as Th17 cells, measured prior to HIV acquisition, were associated with markers of disease progression after infection. We analyzed blood samples from individuals who were HIV-negative at the time of sampling and later acquired HIV during follow-up. We found that individuals with higher pre-infection levels of Th17 cells experienced faster immune decline after HIV infection, including more rapid loss of CD4+ T cells and lower CD4/CD8 ratios. These associations were observed independently of viral load and varied by cohort, age and sex. Our findings indicate that immune conditions present before HIV infection can influence subsequent disease progression and suggest that Th17 cells may serve as biomarkers to identify individuals at higher risk of rapid HIV-related immune damage.
]]></description>
<dc:creator>Omole, T. E.</dc:creator>
<dc:creator>Nguyen, H. M.</dc:creator>
<dc:creator>Marcinow, A.</dc:creator>
<dc:creator>Jahan, N.</dc:creator>
<dc:creator>Severini, G.</dc:creator>
<dc:creator>Naicker, N.</dc:creator>
<dc:creator>Thomas, K.</dc:creator>
<dc:creator>Celum, C.</dc:creator>
<dc:creator>Mugo, N.</dc:creator>
<dc:creator>Mujugira, A.</dc:creator>
<dc:creator>Kublin, J.</dc:creator>
<dc:creator>Corey, L.</dc:creator>
<dc:creator>Sivro, A.</dc:creator>
<dc:creator>Lingappa, J.</dc:creator>
<dc:creator>Gray, G.</dc:creator>
<dc:creator>McKinnon, L.</dc:creator>
<dc:date>2025-12-29</dc:date>
<dc:identifier>doi:10.64898/2025.12.29.696811</dc:identifier>
<dc:title><![CDATA[Pre-Human Immunodeficiency Virus (HIV) infection Th17 CD4+ T cells as predictors of early HIV disease progression]]></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/10.64898/2025.12.29.696916v1?rss=1">
<title>
<![CDATA[
Lysine deficiency within a conserved lysine desert is critical for EEL-1/HUWE1 to support ubiquitin proteasome system function. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.29.696916v1?rss=1</link>
<description><![CDATA[
The ubiquitin proteasome system (UPS) is the primary mechanism for targeted protein degradation in eukaryotic cells. Dysfunction of this system is a driver of human disease and a hallmark of aging and late-onset neurodegenerative disorders. Understanding the mechanisms that ensure robust protein turnover may provide new avenues for treatment in these contexts. E3 ubiquitin ligases play critical roles in supplying ubiquitinated substrates to the proteasome, with HUWE1 being an enormous, versatile, and highly conserved member of this family. Here, we show that the C. elegans HUWE1 ortholog, EEL-1, contributes to robust protein turnover during challenges to the proteolytic capacity of the proteasome. We demonstrate that the ability of EEL-1/HUWE1 to safeguard protein turnover requires ubiquitin-binding domains within the substrate-binding arena and the HECT-type ubiquitin ligase activity, supporting a model in which EEL-1 ensures degradation by increasing ubiquitination of pre-ubiquitinated substrates. EEL-1 contains extensive lysine-deficient regions, found at conserved locations in its substrate-binding arena. Through unbiased mutagenesis screening and precise engineering of the EEL-1 protein, we uncover that introducing lysine residues into these regions is detrimental to UPS function. Together, our findings indicate a central and evolutionarily ancient role for EEL-1/HUWE1 in maintaining optimal UPS function and support targeting this E3 for therapeutic manipulation.
]]></description>
<dc:creator>Yanagi, K. S.</dc:creator>
<dc:creator>Chen, B. J.</dc:creator>
<dc:creator>Kunjo, S. O.</dc:creator>
<dc:creator>Topalidou, I.</dc:creator>
<dc:creator>Lehrbach, N. J.</dc:creator>
<dc:date>2025-12-30</dc:date>
<dc:identifier>doi:10.64898/2025.12.29.696916</dc:identifier>
<dc:title><![CDATA[Lysine deficiency within a conserved lysine desert is critical for EEL-1/HUWE1 to support ubiquitin proteasome system function.]]></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/10.64898/2025.12.30.692369v1?rss=1">
<title>
<![CDATA[
CausalGRN: deciphering causal gene regulatory networks from single-cell CRISPR screens 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.30.692369v1?rss=1</link>
<description><![CDATA[
Large-scale single-cell CRISPR screens with single-cell RNA-seq (scRNA-seq) readouts provide critical data to map causal gene regulatory networks (GRNs). However, translating the complex scRNA-seq outputs into reliable causal insights remains a major analytical challenge. Here we present CausalGRN, a scalable computational framework that infers causal GRNs and predicts cellular responses to unseen perturbations. CausalGRN first mitigates pervasive spurious partial correlations in sparse scRNA-seq data through a novel adaptive thresholding correction, enabling robust inference of an undirected graph. It then orients this graph using observed perturbation outcomes. The resulting directed GRN can be used to predict the downstream effects of novel perturbations via network propagation. Across both simulations and diverse experimental datasets, CausalGRN substantially outperforms existing approaches in network reconstruction accuracy and in predicting the effects of unseen perturbations, providing a principled bridge from perturbation data to causal gene regulation.
]]></description>
<dc:creator>Yu, B.</dc:creator>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Qi, G.</dc:creator>
<dc:creator>Huangfu, D.</dc:creator>
<dc:creator>Hsu, L.</dc:creator>
<dc:creator>Shojaie, A.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2025-12-31</dc:date>
<dc:identifier>doi:10.64898/2025.12.30.692369</dc:identifier>
<dc:title><![CDATA[CausalGRN: deciphering causal gene regulatory networks from single-cell CRISPR screens]]></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/10.64898/2026.01.02.697352v1?rss=1">
<title>
<![CDATA[
Developmental programming of hematopoietic stem cell dormancy by evasion of Notch signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.02.697352v1?rss=1</link>
<description><![CDATA[
Dormant hematopoietic stem cells (HSCs) are a rare subset of deeply quiescent, metabolically inactive cells that preserve lifelong hematopoietic regeneration. In mice, these cells were thought to arise exclusively after birth in the BM, while fetal liver HSCs were considered uniformly proliferative and rapidly expanding. Here, using H2B-GFP label-retention strategies, we identify a previously unrecognized population of low-dividing HSCs that is specified during embryogenesis and persists into adulthood. We show that this dormant state is established in specific fetal HSCs that evade the activation of Notch signaling within the fetal liver niche, rather than through global loss of pathway activity. Genetic and transient perturbation of Notch signaling reveals a narrow developmental window in during which reduced Notch exposure promotes entry into a durable dormancy program, whereas irreversible pathway disruption compromises long-term function. Notably, transient Notch during fetal development, but not in adult BM, induces HSC quiescence while enhancing long-term repopulation capacity. Together, these findings demonstrate that HSC dormancy is developmentally programmed before birth and identify evasion of Notch signaling as a key mechanism establishing this state, fundamentally revising current models of how long-term regenerative potential is specified.
]]></description>
<dc:creator>Herrero-Molinero, P.</dc:creator>
<dc:creator>Canton, E.</dc:creator>
<dc:creator>Maqueda, M.</dc:creator>
<dc:creator>Ruiz-Herguido, C.</dc:creator>
<dc:creator>Iglesias, A.</dc:creator>
<dc:creator>Gonzalez, J.</dc:creator>
<dc:creator>Hadland, B.</dc:creator>
<dc:creator>Espinosa, L.</dc:creator>
<dc:creator>Bigas, A.</dc:creator>
<dc:date>2026-01-04</dc:date>
<dc:identifier>doi:10.64898/2026.01.02.697352</dc:identifier>
<dc:title><![CDATA[Developmental programming of hematopoietic stem cell dormancy by evasion of Notch signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.05.697808v1?rss=1">
<title>
<![CDATA[
Influenza hemagglutinin subtypes have different sequence constraints despite sharing extremely similar structures 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.05.697808v1?rss=1</link>
<description><![CDATA[
Hemagglutinins (HA) from different influenza A virus subtypes share as little as [~]40% amino acid identity, yet their protein structure and cell entry function are highly conserved. Here we examine the extent that sequence constraints on HA differ across three subtypes. To do this, we first use pseudovirus deep mutational scanning to measure how all amino-acid mutations to an H7 HA affect its cell entry function. We then compare these new measurements to previously described measurements of how all mutations to H3 and H5 HAs affect cell entry function. We find that [~]50% of HA sites display substantially diverged preferences for different amino acids across the HA subtypes. The sites with the most divergent amino-acid preferences tend to be buried and have biochemically distinct wildtype amino acids in the different HA subtypes. We provide an example of how rewiring the interactions among contacting residues has dramatically shifted which amino acids are tolerated at specific sites. Overall, our results show how proteins with the same structure and function can become subject to very different site-specific evolutionary constraints as their sequences diverge.
]]></description>
<dc:creator>Ahn, J. J.</dc:creator>
<dc:creator>Yu, T. C.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Radford, C. E.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2026-01-06</dc:date>
<dc:identifier>doi:10.64898/2026.01.05.697808</dc:identifier>
<dc:title><![CDATA[Influenza hemagglutinin subtypes have different sequence constraints despite sharing extremely similar structures]]></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/10.64898/2026.01.13.699341v1?rss=1">
<title>
<![CDATA[
Hypatia: a set of quantitative methods for profiling isoform across cell populations 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.13.699341v1?rss=1</link>
<description><![CDATA[
High-throughput long-read single-cell RNA-sequencing enables isoform-level study across single cells, yet methods for systematically assessing cell-to-cell variations remain limited. Here, we develop Hypatia, a comprehensive platform for dissecting isoform complexities across cell populations, devising Tsallis entropy and Cramers V to facilitate robust comparative profiling. Hypatia revealed prominent isoform species variations and usage shifts across cell-types in glioblastoma, renal cell carcinoma, and heart, highlighting clinically relevant applications for studying isoform-derived, cell-specific functions.
]]></description>
<dc:creator>Pan, T.</dc:creator>
<dc:creator>Shiau, C.-K. S.</dc:creator>
<dc:creator>Lu, L.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Bhimaraj, A.</dc:creator>
<dc:creator>Brat, D. J.</dc:creator>
<dc:creator>Huse, J. T.</dc:creator>
<dc:creator>Li, J. J.</dc:creator>
<dc:creator>Gao, R.</dc:creator>
<dc:date>2026-01-14</dc:date>
<dc:identifier>doi:10.64898/2026.01.13.699341</dc:identifier>
<dc:title><![CDATA[Hypatia: a set of quantitative methods for profiling isoform across cell populations]]></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/10.64898/2026.01.14.699358v1?rss=1">
<title>
<![CDATA[
Unifying phylogenetic traversal and deep learning to guide tree exploration 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.14.699358v1?rss=1</link>
<description><![CDATA[
Deep learning offers hope for more efficient phylogenetic inference methods. However, it has yet to have the transformative effect on phylogenetics that it has had in other fields. Here we present a novel approach that combines deep learning with concepts behind current successful phylogenetic algorithms. Specifically, we give the deep learning algorithm access to the output of a phylogenetic dynamic program on the sequence alignment, rather than the raw sequence alignment. The algorithm then learns features based on these phylogenetically processed versions of the sequence data, which provides information that could inform local tree search. For this paper, our goal is simple: predict for each edge in a tree whether it is in a maximum parsimony tree or not. Our model consists of a recurrent neural network that learns features while traversing the input tree, which are used to classify the edge. The model makes high-quality predictions for this NP-complete problem on simulated and empirical datasets for trees of various sizes, and we believe is a stepping stone towards efficient phylogenetic inference using deep learning.
]]></description>
<dc:creator>Collienne, L.</dc:creator>
<dc:creator>Richman, H.</dc:creator>
<dc:creator>Rich, D. H.</dc:creator>
<dc:creator>Barker, M.</dc:creator>
<dc:creator>Jennings-Shaffer, C.</dc:creator>
<dc:creator>Matsen, F. A.</dc:creator>
<dc:date>2026-01-14</dc:date>
<dc:identifier>doi:10.64898/2026.01.14.699358</dc:identifier>
<dc:title><![CDATA[Unifying phylogenetic traversal and deep learning to guide tree exploration]]></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/10.64898/2026.01.14.699605v1?rss=1">
<title>
<![CDATA[
Ribosome-associated quality control of aberrant protein production during amino acid limitation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.14.699605v1?rss=1</link>
<description><![CDATA[
Amino acids can become limiting for protein synthesis through depletion of charged tRNAs, leading to ribosome stalling and disruption of translation elongation at specific codons. To assess whether this is a mechanism by which amino acid availability can directly influence gene expression, we designed a reporter library to measure translation disruption across all sense codons in the context of amino acid limitations. We found that arginine limitation consistently impairs translation at the arginine codon AGA, resulting in synthesis of proteins from endogenous transcripts. In contrast, GCN2 pathway activation suppresses translation disruption following depletion of most other amino acids. Genome-wide screens revealed that the ribosome quality control trigger (RQC-T) and RQC pathways, which resolve ribosome collisions on defective mRNAs, catalyze ribosome splitting and premature fall-off in response to arginine depletion. Additionally, the E3 ubiquitin ligase RNF14, recently shown to clear ribosome A-site obstructions, promotes translation disruption through both ribosome fall-off and frameshifting during arginine limitation. Together, these data show that the RQC machinery is engaged by tRNA-limited ribosomes and identify a new role for RNF14 as a regulator of translation upon arginine limitation.
]]></description>
<dc:creator>Darnell, A. M.</dc:creator>
<dc:creator>Chidley, C.</dc:creator>
<dc:creator>Paradise, V.</dc:creator>
<dc:creator>Cui, D. S.</dc:creator>
<dc:creator>Davidsen, K.</dc:creator>
<dc:creator>Lincoln, S. C.</dc:creator>
<dc:creator>Abbott, K. L.</dc:creator>
<dc:creator>Elbashir, R.</dc:creator>
<dc:creator>Vander Heiden, C. P.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>Sullivan, L. B.</dc:creator>
<dc:creator>Davis, J. H.</dc:creator>
<dc:creator>Vander Heiden, M. G.</dc:creator>
<dc:date>2026-01-15</dc:date>
<dc:identifier>doi:10.64898/2026.01.14.699605</dc:identifier>
<dc:title><![CDATA[Ribosome-associated quality control of aberrant protein production during amino acid limitation]]></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/10.64898/2026.01.16.699998v1?rss=1">
<title>
<![CDATA[
B Lymphocyte Protein Factories produced by Hematopoietic Stem Cell Gene Editing 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.16.699998v1?rss=1</link>
<description><![CDATA[
Long-term in vivo production of therapeutic proteins and development of vaccines that elicit protective levels of broadly neutralizing antibodies (bNAbs) against major pathogens face challenges. Here we report on an alternative gene-editing approach using small numbers of hematopoietic stem and progenitor cells (HSPCs) to direct long-term, high-level expression of antibodies or cargo proteins. Edited B lymphocyte offspring can be activated by cognate antigen to undergo clonal expansion and develop into specific antibody or cargo protein-synthesizing plasma cells. These cells produce long-lasting, therapeutic levels of serum antibody against HIV-1 or malaria and an anti-influenza virus bNAb that mediated universal protection from heterologous lethal challenge. Our data provide a paradigm for cell therapy approaches to prevent or treat disease using self-amplifying B cell protein factories.
]]></description>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Ruprecht, C.</dc:creator>
<dc:creator>Yao, K.-H.</dc:creator>
<dc:creator>Laffont, P.</dc:creator>
<dc:creator>Lima dos Reis, G.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Hägglöf, T.</dc:creator>
<dc:creator>Binet, L.</dc:creator>
<dc:creator>Loewe, M.</dc:creator>
<dc:creator>Hong, J. P.</dc:creator>
<dc:creator>Xiao, T.</dc:creator>
<dc:creator>Sefik, E.</dc:creator>
<dc:creator>Hernandez, B.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Jankovic, M.</dc:creator>
<dc:creator>Seaman, M. S.</dc:creator>
<dc:creator>Costa, G.</dc:creator>
<dc:creator>Nelson, S. A.</dc:creator>
<dc:creator>Clark, J.</dc:creator>
<dc:creator>Kanatani, S.</dc:creator>
<dc:creator>Wilson, P. C.</dc:creator>
<dc:creator>Krammer, F.</dc:creator>
<dc:creator>Levashina, E. A.</dc:creator>
<dc:creator>Julien, J.-P.</dc:creator>
<dc:creator>Wardemann, H.</dc:creator>
<dc:creator>Sinnis, P.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Flavell, R. A.</dc:creator>
<dc:creator>Nussenzweig, M. C.</dc:creator>
<dc:date>2026-01-18</dc:date>
<dc:identifier>doi:10.64898/2026.01.16.699998</dc:identifier>
<dc:title><![CDATA[B Lymphocyte Protein Factories produced by Hematopoietic Stem Cell Gene Editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.23.701336v1?rss=1">
<title>
<![CDATA[
Kinetochore clustering is mediated by Mps1 phosphorylation of conserved MELT motifs in Stu1 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.23.701336v1?rss=1</link>
<description><![CDATA[
Unattached kinetochores cluster in budding yeast to promote microtubule capture. Mallett et al. dissect the molecular mechanisms of this pathway, showing Mps1 kinase regulates an interaction between Stu1 and Slk19 to drive clustering.

ABSTRACTUnattached kinetochores promote microtubule capture while preventing cell cycle progression during mitosis. The Mps1 kinase controls these events by mediating kinetochore assembly of the fibrous corona in animal cells and by triggering the spindle checkpoint. In budding yeast, which does not assemble a fibrous corona, the Stu1 and Slk19 spindle proteins promote microtubule capture by clustering unattached kinetochores, but the underlying mechanism is unclear. Here, we show that Mps1 controls this pathway. We identify two conserved MELT motifs in Stu1 that are directly phosphorylated by Mps1 to recruit Slk19 and mediate kinetochore clustering. Structural analysis of the Stu1:Slk19 complex reveals long, string-like filaments and offers mechanistic insight into how kinetochores might cluster. Our findings reveal parallels between the Mps1-Stu1-Slk19 pathway and the fibrous corona and suggest the regulation of kinetochore capture is a conserved Mps1 function across eukaryotes.
]]></description>
<dc:creator>Mallet, D. R.</dc:creator>
<dc:creator>Jiang, M.</dc:creator>
<dc:creator>Minnuto, G. M.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2026-01-25</dc:date>
<dc:identifier>doi:10.64898/2026.01.23.701336</dc:identifier>
<dc:title><![CDATA[Kinetochore clustering is mediated by Mps1 phosphorylation of conserved MELT motifs in Stu1]]></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/10.64898/2026.01.25.701611v1?rss=1">
<title>
<![CDATA[
Modeling HIV infection, treatment, rebound, and intervention in human immune organoids 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.25.701611v1?rss=1</link>
<description><![CDATA[
Targeting the HIV-infected reservoir in lymphoid tissues (LT) will be critical to developing a cure for people living with HIV (PLWH). LT explants used to study HIV infection enable the evaluation of human-specific disease progression and treatment response; however, their short lifespan makes it challenging to assess long-term treatment interventions. We therefore established an immune organoid model of HIV infection using human tonsil or spleen cells, demonstrating productive HIV infection and viral integration into CD4+ T cells. Treatment with a protease inhibitor fully suppressed ongoing viral production, with virologic rebound occurring within days of treatment interruption. The transfer of healthy allogeneic NK cells to target the reservoir upon treatment interruption reduced the number of infected cells, intact viral genomes, and production of de novo infectious viral particles. Adoption of this immune organoid platform will accelerate the evaluation of cure-based strategies to eliminate the HIV reservoir in tissues for PLWH.
]]></description>
<dc:creator>Sackey, S. A.</dc:creator>
<dc:creator>Schrum, J. E.</dc:creator>
<dc:creator>Mangalanathan, U. M.</dc:creator>
<dc:creator>Fish, R. S.</dc:creator>
<dc:creator>Sola, E.</dc:creator>
<dc:creator>Selehi, A.</dc:creator>
<dc:creator>Pi, R.</dc:creator>
<dc:creator>Zack, J. A.</dc:creator>
<dc:creator>Kim, J. T.</dc:creator>
<dc:creator>Lehman, D. A.</dc:creator>
<dc:creator>Davis, M. M.</dc:creator>
<dc:creator>Blish, C. A.</dc:creator>
<dc:date>2026-01-27</dc:date>
<dc:identifier>doi:10.64898/2026.01.25.701611</dc:identifier>
<dc:title><![CDATA[Modeling HIV infection, treatment, rebound, and intervention in human immune organoids]]></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/10.64898/2026.01.29.702071v1?rss=1">
<title>
<![CDATA[
Recovery of an 18th Century Rhinovirus Genome through Ancient RNA Isolation of Human Lungs 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.29.702071v1?rss=1</link>
<description><![CDATA[
RNA viruses cause substantial global morbidity, yet their impact prior to the twentieth century remains obscured. While ancient DNA studies have transformed our understanding of past pathogens, ancient RNA (aRNA) isolation is largely restricted to exceptionally preserved samples. Here, we simultaneously recover aDNA and aRNA from non-formalin-fixed human lung specimens and reconstructed an 18th-century Human Rhinovirus (HRV) A genome--the oldest human RNA virus identified to date. The RNA is highly fragmented, with distinctive terminal misincorporations and coverage patterns consistent with double-stranded RNA. Phylogenetic analyses indicate that this historical HRV genome is an extinct lineage related to contemporary genotypes, providing a unique perspective on rhinovirus evolution. These findings demonstrate that centuries-old medical specimens can retain informative aRNA, expanding the temporal scope of paleovirology.
]]></description>
<dc:creator>Barnett, E. E.</dc:creator>
<dc:creator>Castillo, A.</dc:creator>
<dc:creator>Du Plessis, I. A.</dc:creator>
<dc:creator>Kistler, K.</dc:creator>
<dc:creator>Carrillo, L.</dc:creator>
<dc:creator>Leon, A. S.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Rutherford, M. G.</dc:creator>
<dc:creator>Ploug, J.</dc:creator>
<dc:creator>McCrone, J. T.</dc:creator>
<dc:creator>Arcos, M. C. A.</dc:creator>
<dc:creator>Melo, D. B.</dc:creator>
<dc:date>2026-01-30</dc:date>
<dc:identifier>doi:10.64898/2026.01.29.702071</dc:identifier>
<dc:title><![CDATA[Recovery of an 18th Century Rhinovirus Genome through Ancient RNA Isolation of Human Lungs]]></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/10.64898/2026.01.29.702664v1?rss=1">
<title>
<![CDATA[
Shared Maturation Pathways of HIV-1 Envelope-reactive V3-glycan bnAb Lineages in Human and Rhesus Macaque 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.29.702664v1?rss=1</link>
<description><![CDATA[
ABSTRACTUnderstanding broadly neutralizing antibody (bnAb) lineage development in rhesus macaques (RMs) infected with Simian-HIV (SHIV) may inform HIV-1 vaccine designs. We analyzed HIV-1 envelope-antibody coevolution in 18 RMs infected by SHIV.BG505 (subtype A) and found highly conserved patterns of antibody recognition and Env escape, including in three animals that developed V3-glycan bnAbs. From one RM with V3-glycan targeted plasma breadth, we isolated 203 members of a single clonal Ab lineage designated DH1030. DH1030 Abs demonstrated striking genetic, functional and structural similarities with the human V3-glycan bnAb lineage DH270, which was isolated from an individual with subtype C HIV-1 CH848 infection. Notably, human-DH270 and macaque-DH1030 bnAbs shared early improbable mutations in HCDR2 that were critical for bnAb development. Thus, key improbable mutations and convergent patterns of antibody evolution and epitope recognition were shared across primate species and distinct HIV-1 subtypes, findings that may be leveraged in new HIV-1 vaccine designs.

ONE SENTENCE SUMMARYBroadly neutralizing antibodies targeting HIV-1 envelope surface protein develop via a common maturation pathway across primate phylogeny.
]]></description>
<dc:creator>Clark, M.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Martin, M.</dc:creator>
<dc:creator>Evangelous, T.</dc:creator>
<dc:creator>Gobeil, S.</dc:creator>
<dc:creator>Berry, M.</dc:creator>
<dc:creator>Wagh, K.</dc:creator>
<dc:creator>Giorgi, E.</dc:creator>
<dc:creator>Hogarty, M.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Marchitto, L.</dc:creator>
<dc:creator>Ding, W.</dc:creator>
<dc:creator>Park, Y.</dc:creator>
<dc:creator>Rando, J.</dc:creator>
<dc:creator>Carey, J.</dc:creator>
<dc:creator>Connell, A.</dc:creator>
<dc:creator>Skelly, A.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Rohr, S.</dc:creator>
<dc:creator>Jiang, C.</dc:creator>
<dc:creator>Venkatayogi, S.</dc:creator>
<dc:creator>Mansouri, K.</dc:creator>
<dc:creator>Edwards, R. J.</dc:creator>
<dc:creator>Lindenberger, J.</dc:creator>
<dc:creator>Hora, B.</dc:creator>
<dc:creator>Doria-Rose, N. A.</dc:creator>
<dc:creator>Mascola, J.</dc:creator>
<dc:creator>van Gils, M. J.</dc:creator>
<dc:creator>Sanders, R. W.</dc:creator>
<dc:creator>Korber, B.</dc:creator>
<dc:creator>Hahn, B. H.</dc:creator>
<dc:creator>Saunders, K. O.</dc:creator>
<dc:creator>Acharya, P.</dc:creator>
<dc:creator>Wiehe, K.</dc:creator>
<dc:creator>Haynes, B. F.</dc:creator>
<dc:creator>Shaw, G. M.</dc:creator>
<dc:creator>Williams, W. B.</dc:creator>
<dc:date>2026-01-30</dc:date>
<dc:identifier>doi:10.64898/2026.01.29.702664</dc:identifier>
<dc:title><![CDATA[Shared Maturation Pathways of HIV-1 Envelope-reactive V3-glycan bnAb Lineages in Human and Rhesus Macaque]]></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/10.64898/2026.02.03.702568v1?rss=1">
<title>
<![CDATA[
Genetic engineering of Staphylococcus haemolyticus: Overcoming Restriction-Modification Barriers and Targeting Virulence Genes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.03.702568v1?rss=1</link>
<description><![CDATA[
Staphylococcus haemolyticus is an emerging multidrug-resistant nosocomial pathogen noted for robust biofilm formation and complex restriction-modification (RM) systems that hinder genetic manipulation. These barriers have severely limited mechanistic studies into its pathogenesis and immune evasion. Here, we report the development of a molecular toolbox that enables precise genomic engineering of clinical S. haemolyticus isolates. Using PacBio Single Molecule Real Time (SMRT) and bisulfite sequencing, we defined the complete genomes and methylomes of nine isolates, generating a functional readout of the active RM defences present in each strain. Among the RM systems identified, a Type II (PDLC03279) and a Type III (PDL3649/PDLC03643) system were significantly overrepresented in clinical isolates, suggesting a potential role in adaptation to host or hospital-associated environments. To bypass these RM barriers, we implemented a dual strategy: first, applying SyngenicDNA-based approaches to eliminate RM target motifs from genetic tools; and second, engineering a surrogate E. coli strain (JMC4) to mimic conserved S. haemolyticus methylation patterns. These tools significantly enhanced transformation efficiency and enabled targeted knockout of four putative virulence genes (sraP, secA2, capA, and capI), as well as allelic exchange of the native capsule operon with the corresponding region from a non-encapsulated isolate. To our knowledge, this is the first report of precise genomic modifications in S. haemolyticus. The establishment of robust molecular tools for transformation and genome editing lays a foundation for future functional studies of virulence and host adaptation in this resilient opportunistic pathogen.

Data summaryAll genomic data were deposited in the European Nucleotide Archive and are available in NCBI under the project accession number PRJEB2705. Supporting data are provided in supplementary data files. Accession numbers are given in Table 1.

O_TBL View this table:
org.highwire.dtl.DTLVardef@1706c82org.highwire.dtl.DTLVardef@1e40917org.highwire.dtl.DTLVardef@10e7d3org.highwire.dtl.DTLVardef@1e3e64corg.highwire.dtl.DTLVardef@1249b03_HPS_FORMAT_FIGEXP  M_TBL O_FLOATNOTable 1.C_FLOATNO O_TABLECAPTIONS. haemolyticus isolates used in this study.

C_TABLECAPTION C_TBL Impact statementStaphylococcus haemolyticus is a multidrug-resistant opportunistic pathogen which causes disease in vulnerable patients. Previously, genetic modification of this pathogen was prevented due to the prevalence of strain-specific restriction modification systems, which use DNA methylation to identify and foreign DNA (including plasmids) when it enters the cells, and then destroy it using restriction enzymes. We analysed the methylomes of a selection of S. haemolyticus strains and used this data to tailor molecular tools to evade the RM systems. This enabled us to improve transformation efficiency and perform genome editing, including large-scale chromosomal modifications and targeted deletion of suspected virulence genes (a first for S. haemolyticus). Beyond its immediate relevance to researchers studying this pathogen, the tools and approaches developed here have broader utility for genetic engineering of other coagulase-negative staphylococci and bacteria with similar RM barriers. Additionally, we found that two RM systems were enriched in clinical strains, suggesting they may have a function in virulence, in addition to their roles as bacterial phage defence systems.
]]></description>
<dc:creator>Venter, H. J.</dc:creator>
<dc:creator>Cavanagh, J. P.</dc:creator>
<dc:creator>Wolden, R.</dc:creator>
<dc:creator>Jones, D.</dc:creator>
<dc:creator>Roberts, R. J.</dc:creator>
<dc:creator>Rivera, M. Z.</dc:creator>
<dc:creator>Christensen, M. O.</dc:creator>
<dc:creator>Johnston, C. D.</dc:creator>
<dc:date>2026-02-03</dc:date>
<dc:identifier>doi:10.64898/2026.02.03.702568</dc:identifier>
<dc:title><![CDATA[Genetic engineering of Staphylococcus haemolyticus: Overcoming Restriction-Modification Barriers and Targeting Virulence Genes]]></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/10.64898/2026.02.04.703720v1?rss=1">
<title>
<![CDATA[
Intranasal Delivery of HIV/SIV Antigens with NE/AS01B Adjuvants Enhances Cellular Immunity and Reduces Viral Loads in SHIV-Challenged Macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.04.703720v1?rss=1</link>
<description><![CDATA[
The primary route of HIV transmission is across mucosal tissues; therefore, developing a protective mucosal vaccine is a top priority. In a pilot study, using a macaque model, we delivered HIV gp140 envelope glycoprotein and SIVmac239 P55 Gag and Nef antigens using heterologous prime/boost via the intranasal route with a soybean oil-based nanoemulsion (NE) adjuvant and through the intramuscular route with the AS01B adjuvant system to generate enhanced cell-mediated immunity. We used a NE adjuvant to promote gut-homing cell-mediated immunity and the AS01B system to enhance humoral immune responses. Following intrarectal challenge with SHIV 4MTF.tHy, vaccinated macaques acquired the virus but experienced lower viral loads in plasma (P=0.003) and CSF (P=0.001), and potent polyfunctional gag-specific (CD107a+, IFN{gamma}, TNF+) responses across diverse lymph nodes. Significant antibody-dependent complement deposition (ADCD) and antibody-dependent cellular phagocytosis (ADCP) responses were induced, and gut-microbiome crosstalk could be modulated and showing reduced SHIV-dysbiosis. Notably, vaccination preserved mucosal all-trans retinoic acid levels (atRA) (p<0.05). However, no significant differences were observed for antibody responses between vaccinated and unvaccinated macaques. In summary, the induced gut-homing properties by the NE adjuvant are effective at generating cell-mediated immunity and reducing viral set points and warrant further investigations as a mucosal adjuvant in HIV vaccine design.

ImportanceThree major non-mucosal vaccine trials (RV144, HVTN702, and 706) failed to reduce HIV infection rates. Therefore, new approaches in developing a mucosal vaccine remain an effective strategy to attempt to control HIV infection. Coherent vaccine approaches against HIV were focused on immune correlates related to viral loads, persistent reservoirs, and antibody responses. As a proof-of-principle, we developed a vaccine regimen consisting of AS01B and an adjuvanted oil-in-water NE cleaved HIV clade C gp140 protein and non-cleaved Gag, and nef particles administered through intranasal, subcutaneous, and intramuscular routes, followed by intrarectal challenge with clade C SHIV. This vaccine elicited strong ADCD and ADCP responses, modulated immune-microbiome crosstalk, and reduced susceptibility to SHIV-infection-associated dysbiosis. Additionally, it preserved mucosal all trans retinoic acid (atRA) levels, suggesting a potential role for this approach in HIV vaccine development.
]]></description>
<dc:creator>Thurman, M.</dc:creator>
<dc:creator>Chokkavelu, V.</dc:creator>
<dc:creator>Johnson, S. D.</dc:creator>
<dc:creator>Olwenyi, O. A.</dc:creator>
<dc:creator>Kathamuthu, G. R.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Adeniji, S.</dc:creator>
<dc:creator>Hong, K. Y.</dc:creator>
<dc:creator>Johnston, M.</dc:creator>
<dc:creator>Bose, D.</dc:creator>
<dc:creator>Pandey, K.</dc:creator>
<dc:creator>Gao, H.</dc:creator>
<dc:creator>Shen, X.</dc:creator>
<dc:creator>Montefiori, D.</dc:creator>
<dc:creator>Wong, P. T.</dc:creator>
<dc:creator>Baker, J. R.</dc:creator>
<dc:creator>Villinger, F.</dc:creator>
<dc:creator>Kane, M. A.</dc:creator>
<dc:creator>Abdel-Mohsen, M.</dc:creator>
<dc:creator>Byrareddy, S.</dc:creator>
<dc:date>2026-02-05</dc:date>
<dc:identifier>doi:10.64898/2026.02.04.703720</dc:identifier>
<dc:title><![CDATA[Intranasal Delivery of HIV/SIV Antigens with NE/AS01B Adjuvants Enhances Cellular Immunity and Reduces Viral Loads in SHIV-Challenged Macaques]]></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/10.64898/2026.02.07.704521v1?rss=1">
<title>
<![CDATA[
Structural determinants of broadly neutralizing human antibodies binding to morphological dengue virus variants. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.07.704521v1?rss=1</link>
<description><![CDATA[
Dengue virus (DENV) is a global disease threat with tropical regions bearing the bulk of disease burden. Presence of four DENV serotypes with varying antigenicity has complicated development of effective, pan-serotype strategies for prevention and treatment of dengue disease. Here, we report single-particle cryo-electron microscopy (cryo-EM) structures of DENV in complex with three human-derived broadly neutralizing antibodies (bnAbs), revealing an antibody class which preferentially uses its heavy chain for potency and antigen recognition. These heavy chain-driven E-dimer recognizing (HEDR) bnAbs potently neutralize all DENV serotypes by binding to a quaternary epitope encompassing the critical fusion loop and conserved glycans on DENVs surface E-glycoprotein dimer. We also demonstrate the presence of capsule-shaped DENV, with HEDR antibodies binding such tubular morphologies in addition to spherical virions. Cryo-EM helical reconstructions of Fab-bound tubular virions from different DENV strains establishes that the pattern of E-glycoproteins helical arrangement is dependent solely on the virus strain. The structural data also demonstrate that binding of highly potent HEDR bnAb D14.F25.S02 causes distortion of DENV particles. Collectively, these results elucidate key features of HEDR bnAbs while illustrating the importance of eliciting immune responses that can neutralize diverse DENV morphologies.
]]></description>
<dc:creator>Mangala Prasad, V.</dc:creator>
<dc:creator>Chatterjee, A.</dc:creator>
<dc:creator>Roy, A.</dc:creator>
<dc:creator>Srinivasan, S.</dc:creator>
<dc:creator>Charles, S.</dc:creator>
<dc:creator>Lubow, J.</dc:creator>
<dc:creator>goo, l.</dc:creator>
<dc:date>2026-02-08</dc:date>
<dc:identifier>doi:10.64898/2026.02.07.704521</dc:identifier>
<dc:title><![CDATA[Structural determinants of broadly neutralizing human antibodies binding to morphological dengue virus variants.]]></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/10.64898/2026.02.10.705188v1?rss=1">
<title>
<![CDATA[
Deep learning-based non-invasive profiling of tumor transcriptomes from cell-free DNA for precision oncology 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.10.705188v1?rss=1</link>
<description><![CDATA[
Circulating tumor DNA (ctDNA) profiling from liquid biopsies is increasingly adopted as a minimally invasive solution for clinical cancer diagnostic applications. Current methods for inferring gene expression from ctDNA require specialized assays or ultra-deep, targeted sequencing, which preclude transcriptome-wide profiling at single-gene resolution. Herein we jointly introduce Triton, a tool for comprehensive fragmentomic and nucleosome profiling of cell-free DNA (cfDNA), and Proteus, a multi-modal deep learning framework for predicting single gene expression, using standard depth ([~]30-120x) whole genome sequencing of cfDNA. By synthesizing fragmentation and inferred nucleosome positioning patterns in the promoter and gene body from Triton, Proteus reproduced expression profiles using pure ctDNA from patient-derived xenografts (PDX) with an accuracy similar to RNA-Seq technical replicates. Applying Proteus to cfDNA from four patient cohorts with matched tumor RNA-Seq, we show that the model accurately predicted the expression of specific prognostic and phenotype markers and therapeutic targets. As an analog to RNA-Seq, we further confirmed the immediate applicability of Proteus to existing tools through accurate prediction of gene pathway enrichment scores. Our results demonstrate the potential clinical utility of Triton and Proteus as non-invasive tools for precision oncology applications such as cancer monitoring and therapeutic guidance.

SubjectsCirculating tumor DNA, liquid biopsies, patient-derived xenografts, whole genome sequencing, deep learning, convolutional neural network, gene expression
]]></description>
<dc:creator>Patton, R. D.</dc:creator>
<dc:creator>Netzley, A.</dc:creator>
<dc:creator>Persse, T. W.</dc:creator>
<dc:creator>Nair, A.</dc:creator>
<dc:creator>Galipeau, P. C.</dc:creator>
<dc:creator>Coleman, I. M.</dc:creator>
<dc:creator>Itagi, P.</dc:creator>
<dc:creator>Chandra, P.</dc:creator>
<dc:creator>Adil, M.</dc:creator>
<dc:creator>Vashisth, M.</dc:creator>
<dc:creator>Sayar, E.</dc:creator>
<dc:creator>Hiatt, J. B.</dc:creator>
<dc:creator>Dumpit, R.</dc:creator>
<dc:creator>Kollath, L.</dc:creator>
<dc:creator>Demirci, R. A.</dc:creator>
<dc:creator>Ghodsi, A.</dc:creator>
<dc:creator>Lam, H.-M.</dc:creator>
<dc:creator>Morrissey, C.</dc:creator>
<dc:creator>Iravani, A.</dc:creator>
<dc:creator>Chen, D. L.</dc:creator>
<dc:creator>Hsieh, A. C.</dc:creator>
<dc:creator>MacPherson, D.</dc:creator>
<dc:creator>Haffner, M. C.</dc:creator>
<dc:creator>Nelson, P. S.</dc:creator>
<dc:creator>Ha, G.</dc:creator>
<dc:date>2026-02-12</dc:date>
<dc:identifier>doi:10.64898/2026.02.10.705188</dc:identifier>
<dc:title><![CDATA[Deep learning-based non-invasive profiling of tumor transcriptomes from cell-free DNA for precision oncology]]></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/10.64898/2026.02.12.705519v1?rss=1">
<title>
<![CDATA[
Complete definition of how mutations affect antibodies used to prevent RSV 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.12.705519v1?rss=1</link>
<description><![CDATA[
New antibodies targeting the F protein of respiratory syncytial virus (RSV) have substantially reduced infant hospitalizations. However, viral resistance is a concern: one antibody failed clinical trials due to emergence of a resistant strain, and sporadic resistance mutations to the most widely used antibody (nirsevimab) have been identified in breakthrough infections. Here we define how RSV F mutations affect antibody neutralization. We first provide a biophysical model of how the buffering effect of bivalent IgG binding combines with differences in monovalent Fab potency to explain why nirsevimab resistance mutations are more common in subtype B than subtype A RSV strains. We then perform pseudovirus deep mutational scanning to safely measure how nearly all mutations to F affect its cell entry function and neutralization by the IgG and Fab forms of nirsevimab, clesrovimab, and several other key antibodies. We use these measurements to enable real-time surveillance of RSV sequences for antibody resistance, and identify rare strains with sporadic resistance mutations. Overall, our work improves understanding of the mechanisms by which viral mutations impact antibody neutralization, enables monitoring for natural RSV strains resistant to antibodies of public-health importance, and can help guide development of future antibodies with resilience to viral escape.
]]></description>
<dc:creator>Simonich, C. A.</dc:creator>
<dc:creator>McMahon, T. E.</dc:creator>
<dc:creator>Kampman, L.</dc:creator>
<dc:creator>Chu, H. Y.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2026-02-12</dc:date>
<dc:identifier>doi:10.64898/2026.02.12.705519</dc:identifier>
<dc:title><![CDATA[Complete definition of how mutations affect antibodies used to prevent RSV]]></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/10.64898/2026.02.11.705165v1?rss=1">
<title>
<![CDATA[
Intellectual disability risk gene RFX4 regulates cortical neurogenesis by restraining neuronal differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.11.705165v1?rss=1</link>
<description><![CDATA[
Despite the recent identification of RFX4 as a neurodevelopmental disorder risk gene, its role in cortical development remained unclear. Here, we identified both shared and lineage-specific RFX4 requirements for human cortical development using new human stem cell models of deficiency and pathogenic mutation. We found that RFX4 restrains neurogenesis by acting cooperatively with NOTCH signaling, specifically repressing pro-neuronal and synaptic gene expression in neural progenitors. We also determined that genome-wide binding of RFX3, another neurodevelopmental disorder risk gene, depends upon RFX4 to regulate synaptic gene expression. Furthermore, we identified lineage-specific functions for RFX4 in regulating proliferation during cortical inhibitory neuron development. Ultimately, we demonstrated that RFX4 deficiency persistently dysregulates neuronal gene expression through neuronal differentiation and disrupts cortical neuron stratification in organoid models. These consequences were absent in neurons generated by direct differentiation, confirming that neuronal phenotypes resulted from unconstrained neurogenesis. Finally, we modeled pathogenic missense mutation of the RFX4 DNA-binding domain. While this mutation strongly reduced DNA binding, it dysregulated synaptic gene expression distinctly from our deficiency models, supporting pathogenic mechanisms distinct from haploinsufficiency. Together, this work identified both shared and lineage-specific requirements for RFX4 during cortical development, building a necessary foundation for elucidating the etiology of RFX4-associated disorders.
]]></description>
<dc:creator>Kroll, K. L.</dc:creator>
<dc:creator>Determan, J. J.</dc:creator>
<dc:creator>Chapman, G.</dc:creator>
<dc:creator>Crump, S. R.</dc:creator>
<dc:creator>Batool, F.</dc:creator>
<dc:creator>Malik, S.</dc:creator>
<dc:creator>Gujral, T.</dc:creator>
<dc:creator>Buchser, W.</dc:creator>
<dc:creator>Valentine, C.</dc:creator>
<dc:creator>Elia, S.</dc:creator>
<dc:creator>Sentmanat, M.</dc:creator>
<dc:creator>Jetter, H.</dc:creator>
<dc:date>2026-02-14</dc:date>
<dc:identifier>doi:10.64898/2026.02.11.705165</dc:identifier>
<dc:title><![CDATA[Intellectual disability risk gene RFX4 regulates cortical neurogenesis by restraining neuronal differentiation]]></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/10.64898/2026.02.12.705483v1?rss=1">
<title>
<![CDATA[
Vicious circle of amyloid and leptomeningeal macrophages evokes vascular dysfunction in CAA 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.12.705483v1?rss=1</link>
<description><![CDATA[
The subarachnoid space contains leptomeningeal arteries and resident immune cells known as subarachnoid macrophages (SAMs). In Alzheimers disease (AD) and related cerebral amyloid angiopathy, amyloid-{beta} (A{beta}) frequently accumulates around the leptomeningeal arteries. Nevertheless, the local immune responses and their functional consequences remain poorly understood. Using longitudinal intravital imaging in a mouse model of AD, we tracked A{beta} deposition and identified a distinct SAM population that migrates to and phagocytoses these arterial A{beta} deposits. SAM recruitment correlated with vascular remodeling, including smooth muscle loss, aneurysm formation, and reduced cerebral perfusion. Transcriptomic profiling revealed a distinct SAM population in AD with upregulated CD39 expression. Pharmacological inhibition of CD39 attenuated arterial A{beta} deposition, identifying SAMs as a potential therapeutic target in AD.
]]></description>
<dc:creator>Wake, H.</dc:creator>
<dc:creator>Hartantyo, R. Y.</dc:creator>
<dc:creator>TSUJI, T.</dc:creator>
<dc:creator>Hou, L.</dc:creator>
<dc:creator>Saito, R.</dc:creator>
<dc:creator>Yamasaki, A.</dc:creator>
<dc:creator>Kochi, T.</dc:creator>
<dc:creator>Saito, Y.</dc:creator>
<dc:creator>Tada, M.</dc:creator>
<dc:creator>Cheung, D. L.</dc:creator>
<dc:creator>Moorhouse, A. J.</dc:creator>
<dc:creator>Prinz, M.</dc:creator>
<dc:creator>Masuda, T.</dc:creator>
<dc:creator>Kakita, A.</dc:creator>
<dc:date>2026-02-14</dc:date>
<dc:identifier>doi:10.64898/2026.02.12.705483</dc:identifier>
<dc:title><![CDATA[Vicious circle of amyloid and leptomeningeal macrophages evokes vascular dysfunction in CAA]]></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/10.64898/2026.02.18.706711v1?rss=1">
<title>
<![CDATA[
Near real-time data on the human neutralizing antibody landscape to influenza virus as of early 2026 to inform vaccine-strain selection 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.18.706711v1?rss=1</link>
<description><![CDATA[
Twice each year, a decision is made on whether to update the strains included in the seasonal influenza vaccine to better match the most recent circulating viral strains. To characterize the antigenic properties of current seasonal influenza A strains to inform the upcoming decision about which strains to include in the 2026-2027 Northern Hemisphere vaccine, here we perform high-throughput sequencing-based neutralization assays using a library of 57 H3N2 and 34 H1N1 influenza hemagglutinins reflecting the circulating diversity of strains in late 2025 to early 2026. We assay this library against 302 human sera collected in late 2025. The resulting data set encompasses 27,409 titers, and provides a near real-time portrait of the human neutralizing antibody landscape against influenza virus. We find that many human sera have lower titers against the K subclade of H3N2 and the D.3.1.1 subclade of H1N1; these subclades have recently become dominant among their respective subtypes. Our measurements also reveal variability in titers to different subvariants within the K subclade of H3N2, with titers especially low to subclade K strains with additional mutations in antigenic regions D and E. We make all our data and accompanying visualizations publicly available to enable their use in vaccine-strain selection and analyses of influenza evolution and immunity.
]]></description>
<dc:creator>Kikawa, C.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Turner, S. A.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Gang, S.</dc:creator>
<dc:creator>Griffiths, T.</dc:creator>
<dc:creator>Drapeau, E. M.</dc:creator>
<dc:creator>Cowling, B. J.</dc:creator>
<dc:creator>Ho, F.</dc:creator>
<dc:creator>Leung, N. H.</dc:creator>
<dc:creator>Englund, J. A.</dc:creator>
<dc:creator>Lacombe, K.</dc:creator>
<dc:creator>Watanabe, S.</dc:creator>
<dc:creator>Hasegawa, H.</dc:creator>
<dc:creator>Busch, M.</dc:creator>
<dc:creator>Lanteri, M.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Spencer, B.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:creator>Smith, D. J.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Hensley, S. E.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2026-02-19</dc:date>
<dc:identifier>doi:10.64898/2026.02.18.706711</dc:identifier>
<dc:title><![CDATA[Near real-time data on the human neutralizing antibody landscape to influenza virus as of early 2026 to inform vaccine-strain selection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.19.706769v1?rss=1">
<title>
<![CDATA[
Differential effects of two common GVHD prophylaxis regimens on the gut microbiome: Results from the BMT CTN 1801 study 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.19.706769v1?rss=1</link>
<description><![CDATA[
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment for many hematological malignancies, but graft-versus-host disease (GVHD) is a common complication. Low gut microbiome diversity is associated with higher GVHD risk and shorter survival in multiple studies.

Recently, the BMT CTN 1703 clinical trial demonstrated superiority of a GVHD-prophylaxis regimen including post-transplant cyclophosphamide (PTCy) compared to the standard prophylaxis (tacrolimus and methotrexate, Tac/MTX) in terms of GVHD-free, relapse-free survival at one year among reduced intensity conditioning allo-HCT recipients. However, the effect of PTCy on the gut microbiome and its association with clinical outcome have not been described. Here, we report on a companion randomized clinical controlled trial (BMT CTN 1801), which collected 2575 longitudinal stool samples from 304 study participants. Samples were obtained up to weekly up to day 84 post allo-HCT and at less frequent intervals thereafter, up to 2 years.

Microbiome diversity and absolute microbial load were lower in the PTCy group compared to the Tac/MTX group on days 14-28 post-HCT. However, diversity at the timepoint closest to neutrophil engraftment was not significantly associated with non-relapse mortality after one year or other clinical outcomes, contrary to expectations from previous studies. Microbial domination events, when a single species exceeds 30% relative abundance, were comparable across treatment arms and reflected both pathogen blooms as well as less severe disruptions of the microbial community. Clostridium scindens and secondary bile acid metabolism pathways were less prevalent in the PTCy arm than in the Tac/MTX arm post-HCT, yet presence of secondary bile acid metabolism pathways was associated with a lower risk of chronic GVHD.

Given that PTCy was associated with a greater disruption of the microbiome as measured by diversity, absolute microbial abundance, and bile acid metabolism capability, but better clinical outcomes overall, these data suggest that the importance of the microbiome in modulating the host immune systems after allo-HCT is specific to different types of GVHD prophylaxis.
]]></description>
<dc:creator>Wirbel, J.</dc:creator>
<dc:creator>Saber, W.</dc:creator>
<dc:creator>Martens, M. J.</dc:creator>
<dc:creator>Peled, J. U.</dc:creator>
<dc:creator>Andermann, T. M.</dc:creator>
<dc:creator>Fei, T.</dc:creator>
<dc:creator>Brooks, E. F.</dc:creator>
<dc:creator>Doyle, B.</dc:creator>
<dc:creator>Pincus, N. B.</dc:creator>
<dc:creator>Jenq, R. R.</dc:creator>
<dc:creator>Bar, M.</dc:creator>
<dc:creator>Bolanos-Meade, J.</dc:creator>
<dc:creator>Bratrude, B.</dc:creator>
<dc:creator>Chhabra, S.</dc:creator>
<dc:creator>Choi, S. W.</dc:creator>
<dc:creator>Clark, W.</dc:creator>
<dc:creator>Das, S.</dc:creator>
<dc:creator>Elmariah, H.</dc:creator>
<dc:creator>Gooptu, M.</dc:creator>
<dc:creator>Holtan, S. G.</dc:creator>
<dc:creator>Jones, R. J.</dc:creator>
<dc:creator>Levine, J. E.</dc:creator>
<dc:creator>Logan, B. R.</dc:creator>
<dc:creator>Al Malki, M. M.</dc:creator>
<dc:creator>Murthy, H. S.</dc:creator>
<dc:creator>Rashidi, A.</dc:creator>
<dc:creator>Rezvani, A. R.</dc:creator>
<dc:creator>Riches, M. L.</dc:creator>
<dc:creator>Runaas, L.</dc:creator>
<dc:creator>Sandhu, K.</dc:creator>
<dc:creator>Spahn, A.</dc:creator>
<dc:creator>Sung, A. D.</dc:creator>
<dc:creator>van den Brink, M. R. M.</dc:creator>
<dc:creator>Horowitz, M. M.</dc:creator>
<dc:creator>Hamadani, M.</dc:creator>
<dc:creator>Kean, L. S.</dc:creator>
<dc:creator>Perales, M.-A.</dc:creator>
<dc:creator>Bhatt, A. S.</dc:creator>
<dc:date>2026-02-20</dc:date>
<dc:identifier>doi:10.64898/2026.02.19.706769</dc:identifier>
<dc:title><![CDATA[Differential effects of two common GVHD prophylaxis regimens on the gut microbiome: Results from the BMT CTN 1801 study]]></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/10.64898/2026.02.22.707297v1?rss=1">
<title>
<![CDATA[
Mutations to the HCoV-229E spike have counterbalancing effects on serum antibody neutralization and receptor binding 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.22.707297v1?rss=1</link>
<description><![CDATA[
Human coronavirus 229E (HCoV-229E) is an endemic pathogen that causes repeated "common-cold" infections throughout life. Like other coronaviruses, it accumulates spike mutations that erode antibody immunity and enable reinfection. Here, we use pseudovirus deep mutational scanning to measure how mutations to the HCoV-229E spike affect its cell entry function, binding to its human APN receptor, and neutralization by human sera with a range of sensitivities to erosion by viral evolution. We find that both receptor binding and serum neutralization are affected by mutations across spike, including many that modulate these properties by affecting the balance of up versus down conformations of the spike receptor-binding domain (RBD). In particular, some mutations increase both receptor binding and serum neutralization by shifting the RBD to a more up conformation, suggesting that the HCoV-229E spike has evolved to shield key RBD neutralizing epitopes at the cost of less efficient receptor binding.
]]></description>
<dc:creator>Harari, S.</dc:creator>
<dc:creator>Eguia, R. T.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Radford, C. E.</dc:creator>
<dc:creator>Stewart, C.</dc:creator>
<dc:creator>Veesler, D.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2026-02-23</dc:date>
<dc:identifier>doi:10.64898/2026.02.22.707297</dc:identifier>
<dc:title><![CDATA[Mutations to the HCoV-229E spike have counterbalancing effects on serum antibody neutralization and receptor binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.23.707528v1?rss=1">
<title>
<![CDATA[
Strain-specific differences in the response to egg-derived versus recombinant protein influenza vaccines 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.23.707528v1?rss=1</link>
<description><![CDATA[
The 2023/2024 influenza vaccine included an updated H1N1 component designed to better match a new clade of H1N1 that had multiple mutations in antigenic epitopes of hemagglutinin. Despite this update, the vaccine trended towards being less effective against the vaccine-matched H1N1 clade than the parental H1N1 clade lacking the new antigenic mutations. Here we measure neutralization titers of serum antibodies from individuals who had received either a recombinant protein or an egg-derived vaccine against a set of viruses with hemagglutinins from 58 H1N1 strains representative of the diversity during the 2023/2024 season. We find that egg-derived vaccine recipients, but not recombinant protein vaccine recipients, had a relatively lower boost in neutralizing titers to the new clade that the updated vaccine was designed to target. We suggest that the difference in the extent that the egg-derived versus recombinant protein vaccines boost neutralizing titers to the new H1N1 clade is because the seed strain for the egg-derived vaccine strain had acquired a reversion of a key antigenic mutation (K142R) present in that clade. Our results show how egg-derived versus recombinant protein vaccines can elicit different relative titer boosts against different subsets of viral strains, a phenomenon that could impact vaccine effectiveness.

ImportanceInfluenza vaccines can be produced from virus grown in eggs, or grown in cells or made with recombinant protein. Egg-derived influenza vaccines often contain egg-adaptive mutations in the viral antigen hemagglutinin (HA) which can impact the antigenicity or immunogenicity of the HA. In this study, we compare neutralization titers from egg-derived and recombinant protein vaccine recipients against recently circulating influenza A(H1N1) strains. We find that the egg-derived vaccine induces less of a boost in titers than the recombinant protein vaccine to the new clade of viral strains that the vaccine was designed to target.
]]></description>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Tarabi, R. A.</dc:creator>
<dc:creator>Li, S. H.</dc:creator>
<dc:creator>Atkinson, R. K.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>Kikawa, C.</dc:creator>
<dc:creator>Griffths, T.</dc:creator>
<dc:creator>Drapeau, E. M.</dc:creator>
<dc:creator>Wong, S.-S.</dc:creator>
<dc:creator>Cheng, S. M.</dc:creator>
<dc:creator>Leung, N. H.</dc:creator>
<dc:creator>Cobey, S.</dc:creator>
<dc:creator>Cowling, B. J.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:creator>Hensley, S. E.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:date>2026-02-23</dc:date>
<dc:identifier>doi:10.64898/2026.02.23.707528</dc:identifier>
<dc:title><![CDATA[Strain-specific differences in the response to egg-derived versus recombinant protein influenza vaccines]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.23.707578v1?rss=1">
<title>
<![CDATA[
scDesignPop generates realistic population-scale single-cell RNA-seq for power analysis, benchmarking, and privacy protection 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.23.707578v1?rss=1</link>
<description><![CDATA[
Single-cell RNA sequencing (scRNA-seq) combined with genotyping in large cohorts has enabled the discovery of genetic associations with molecular traits (e.g., eQTLs) at cell-type resolution. However, generating population-scale data remains cost-prohibitive, selecting appropriate analysis methods lacks consensus, and sharing eQTL results alongside scRNA-seq data raises privacy risks. To address these challenges, we introduce scDesignPop, a flexible statistical simulator for generating realistic population-scale scRNA-seq data with genetic effects. scDesignPop models cell- and individual-level covariates, putative cell-type-specific eQTLs (cts-eQTLs), and either real or synthetic genotypes. We validated scDesignPop using the OneK1K and CLUES cohorts across 4 qualitative and 16 quantitative metrics. Unlike splatPop, the only existing population-scale simulator, scDesignPop better preserves eQTL effects and gene-gene dependencies within cell types, closely recapitulating characteristics of the reference data. Leveraging its generative framework, scDesignPop enables power analysis in cell types under multiple eQTL model specifications to guide experimental design; facilitates benchmarking of single-cell eQTL mapping methods through user-defined ground truths; and mitigates re-identification risk using synthetic data while retaining cts-eQTL effects.
]]></description>
<dc:creator>Dong, C. Y.</dc:creator>
<dc:creator>Cen, Y.</dc:creator>
<dc:creator>Song, D.</dc:creator>
<dc:creator>Li, J. J.</dc:creator>
<dc:date>2026-02-25</dc:date>
<dc:identifier>doi:10.64898/2026.02.23.707578</dc:identifier>
<dc:title><![CDATA[scDesignPop generates realistic population-scale single-cell RNA-seq for power analysis, benchmarking, and privacy protection]]></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/10.64898/2026.02.25.707521v1?rss=1">
<title>
<![CDATA[
Topological Data Analysis of Spatial Protein Expression in Multiplexed Spatial Proteomics Studies 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.25.707521v1?rss=1</link>
<description><![CDATA[
Multiplexed spatial proteomics platforms generate high-resolution images capturing the spatial expression of proteins in tissue. Images are often fed through a complex pre-processing pipeline to identify individual cells (termed segmentation) and then to predict their phenotypes. It is common to test if the inferred spatial arrangement of cells associates with patient-level outcomes. However, cell segmentation and phenotyping are prone to error and this approach neglects the measured protein levels. Further, new research suggests topological analysis of spatial proteomics may yield more power than alternative approaches. We propose a method, TOASTER, that circumvents reliance on segmentation and phenotyping and instead tests the association between continuous spatial protein expression and a patient-level response variable. TOASTER uses topological data analysis to first characterize the presence of topological features within univariate and bivariate spatial protein expression. The topological structure is summarized using an adaptation of the Nelson-Aalen cumulative hazard function. We can then associate this summary with an outcome using either a functional data analytic approach, a gridwise testing approach, or using kernel association testing. We show via simulation that our approach improves power and controls type I error, even in the presence of gaps or tears in the image which may arise during tissue handling. We apply our approach to a study in triple-negative breast cancer and demonstrate topological features of protein expression associated with immunotherapy response.
]]></description>
<dc:creator>Samorodnitsky, S. N.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:date>2026-02-27</dc:date>
<dc:identifier>doi:10.64898/2026.02.25.707521</dc:identifier>
<dc:title><![CDATA[Topological Data Analysis of Spatial Protein Expression in Multiplexed Spatial Proteomics Studies]]></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/10.64898/2026.02.25.708034v1?rss=1">
<title>
<![CDATA[
Regulatory T cells restrain IL-15-mediated cytotoxic and bystander T cell activity in mucosal tissue without compromising antigen-driven memory 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.25.708034v1?rss=1</link>
<description><![CDATA[
ABSTRACT/SUMMARYMany pathogenic human infections enter the host via a mucosal surface. These nonlymphoid tissues are abundantly populated by polyclonal memory CD8 T cells that persist following infections to protect the host in the event of repeat exposure. Memory T cells can be triggered via T cell receptor (TCR) interaction with their cognate antigen upon re-infection to exert effector functions, including cytotoxicity and cytokine production, and assist in pathogen elimination. Alternatively, some T cells are  bystander activated by cytokines without antigenic signal. This layered approach boosts efccient pathogen clearance but also poses a threat to host tissues if this response is not properly controlled. Here we investigate the regulatory mechanisms modulating the tissue memory CD8 T cell response upon recall, leveraging mouse models to distinguish antigen-driven versus cytokine-activated memory tissue CD8 T cell immunity. We cnd that regulatory T cells (Treg) play a role in restricting cytotoxic and bystander activity in mucosal T cells without compromising the antigen-driven protective memory CD8 T cell response. Critically, Treg provide extrinsic regulation of tissue CD8 T cell cytotoxicity through restriction of available IL-2 and IL-15 trans-presentation. Our cndings help to decne the extrinsic environmental and cellular cues in mucosal tissues that direct tissue memory CD8 T cell cytotoxicity.
]]></description>
<dc:creator>Cruz Talavera, I.</dc:creator>
<dc:creator>Graham, J. B.</dc:creator>
<dc:creator>Swarts, J. L.</dc:creator>
<dc:creator>Traxinger, B. R.</dc:creator>
<dc:creator>Peters, M. Q.</dc:creator>
<dc:creator>Warrier, L.</dc:creator>
<dc:creator>Koehne, A. L.</dc:creator>
<dc:creator>Arkatkar, T.</dc:creator>
<dc:creator>Jerome, K. R.</dc:creator>
<dc:creator>Prlic, M.</dc:creator>
<dc:creator>Lund, J. M.</dc:creator>
<dc:date>2026-02-27</dc:date>
<dc:identifier>doi:10.64898/2026.02.25.708034</dc:identifier>
<dc:title><![CDATA[Regulatory T cells restrain IL-15-mediated cytotoxic and bystander T cell activity in mucosal tissue without compromising antigen-driven memory]]></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/10.64898/2026.03.03.709308v1?rss=1">
<title>
<![CDATA[
Evaluation of an H5 influenza virus mRNA-lipid nanoparticle (LNP) vaccine in lactating dairy cows 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.03.709308v1?rss=1</link>
<description><![CDATA[
Highly pathogenic avian influenza (HPAI) clade 2.3.4.4b H5N1 virus has recently emerged in dairy cattle in the United States. The virus replicates primarily in the mammary gland of infected cattle, leading to dramatic reductions in milk production. It is thought that the virus transmits from animal to animal through viral shedding in milk, and therefore, vaccines that decrease the amount of virus in milk can potentially limit the current outbreak and reduce the risk of H5N1 spillover into humans. Here, we assess the immunogenicity and efficacy of a clade 2.3.4.4b H5 mRNA-LNP vaccine in lactating dairy cows. We found that the H5 mRNA-LNP vaccine elicited robust antibody responses in sera and milk and significantly reduced viral replication and disease caused by clade 2.3.4.4b H5N1 intramammary infection.
]]></description>
<dc:creator>Santos, J. J. S.</dc:creator>
<dc:creator>Souza, C. K.</dc:creator>
<dc:creator>Zanella, G. C.</dc:creator>
<dc:creator>Goulart, D. B.</dc:creator>
<dc:creator>Arruda, B.</dc:creator>
<dc:creator>Boggiatto, P.</dc:creator>
<dc:creator>Palmer, M. V.</dc:creator>
<dc:creator>Snyder, C. A.</dc:creator>
<dc:creator>Kristula, M. A.</dc:creator>
<dc:creator>Dickens, C.</dc:creator>
<dc:creator>Webb, T. L.</dc:creator>
<dc:creator>Atkinson, R. K.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Dwivedi, G.</dc:creator>
<dc:creator>Alameh, M.-G.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Weissman, D.</dc:creator>
<dc:creator>Althouse, G. C.</dc:creator>
<dc:creator>Baker, A. L.</dc:creator>
<dc:creator>Hensley, S. E.</dc:creator>
<dc:date>2026-03-04</dc:date>
<dc:identifier>doi:10.64898/2026.03.03.709308</dc:identifier>
<dc:title><![CDATA[Evaluation of an H5 influenza virus mRNA-lipid nanoparticle (LNP) vaccine in lactating dairy cows]]></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/10.64898/2026.03.03.709191v1?rss=1">
<title>
<![CDATA[
Neuroepithelial reprogramming and ERBB vulnerability in canine acanthomatous ameloblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.03.709191v1?rss=1</link>
<description><![CDATA[
Canine acanthomatous ameloblastoma (CAA) is a locally invasive benign oral neoplasm that is difficult to distinguish from canine oral squamous cell carcinoma (COSCC) due to overlapping clinical, radiologic, and histologic features. Although both tumors exhibit MAPK pathway activation, their mutational landscapes are distinct. Furthermore, previous studies using bulk RNA sequencing (RNA-seq) have demonstrated pronounced differences in programs related to, among others, hypoxia, PI3K-AKT signaling, and cell proliferation. However, these bulk studies lacked the resolution to elucidate the cellular heterogeneity of CAA relative to COSCC. We therefore performed single-nucleus RNA-seq to define the cellular gene expression landscape of CAA, COSCC, and healthy gingiva. Across [~]205,000 nuclei, we identified major epithelial, immune, endothelial, and mesenchymal populations, as well as two epithelial subtypes uniquely enriched in CAA. The CAA-specific keratinocytes exhibited a neuronal-like expression program defined by synaptic regulators, KRAS-associated signaling pathways, and markedly elevated expression of PEG3, ERBB4, GABRB1, MAGI2, and CASK. These findings were validated by bulk RNA-seq, qPCR, and immunohistochemistry, which demonstrated strong nuclear localization of PEG3 exclusively in CAA epithelium. A kinase inhibitor screen independently identified ERBB4 as a candidate therapeutic vulnerability, and pharmacologic inhibition with neratinib was effective. Together, these findings reveal a previously unrecognized neuroepithelial cell state that defines CAA, distinguishes it from COSCC, and reveals unique diagnostic and therapeutic signaling dependencies. Given the molecular and histopathologic parallels between CAA and human ameloblastoma, these data further position CAA as a naturally occurring comparative model for studying ameloblastoma biology and therapeutic vulnerabilities.

Significance StatementCanine acanthomatous ameloblastoma (CAA) is a common locally aggressive oral neoplasm often misdiagnosed as canine oral squamous cell carcinoma (COSCC) due to overlapping clinical, radiologic, and histologic features. Using single-nucleus RNA sequencing of CAA, COSCC, and healthy gingiva, we resolve the microenvironment of these tumors and detect two epithelial subpopulations unique to CAA. These CAA-specific keratinocytes exhibit a neuroepithelial-like signature characterized by synaptic regulators, KRAS-associated signaling, and elevated PEG3 and ERBB4 expression. Functional kinase inhibitor screening of patient derived microtumor slices independently converged on ERBB4 as an effective therapeutic target. Together, our study provides a molecular framework for distinguishing CAA from COSCC and establishes a comparative oncology model with translational relevance to human ameloblastoma and other ERBB-driven epithelial neoplasms.
]]></description>
<dc:creator>Stephanou, A.</dc:creator>
<dc:creator>Shui, B.</dc:creator>
<dc:creator>Mische, D.</dc:creator>
<dc:creator>Byron, M.</dc:creator>
<dc:creator>Katt, W. P.</dc:creator>
<dc:creator>Chan, M.</dc:creator>
<dc:creator>Grenier, J. K.</dc:creator>
<dc:creator>De Vlaminck, I.</dc:creator>
<dc:creator>Duhamel, G. E.</dc:creator>
<dc:creator>Gujral, T. S.</dc:creator>
<dc:creator>Sethupathy, P.</dc:creator>
<dc:creator>Peralta, S.</dc:creator>
<dc:date>2026-03-05</dc:date>
<dc:identifier>doi:10.64898/2026.03.03.709191</dc:identifier>
<dc:title><![CDATA[Neuroepithelial reprogramming and ERBB vulnerability in canine acanthomatous ameloblastoma]]></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/10.64898/2026.03.04.709615v1?rss=1">
<title>
<![CDATA[
Remote homology and functional genetics unmask deeply preserved Scm3/HJURP orthologs in metazoans 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.04.709615v1?rss=1</link>
<description><![CDATA[
In most animals and fungi, centromere identity and function depend on the Scm3/HJURP chaperone, which deposits CENPA at centromeres. However, Scm3/HJURP orthologs appeared to be missing in insects, nematodes, many vertebrates, and other metazoans, suggesting radical chaperone replacement in these lineages. Here, we combine remote homology detection, AlphaFold-based structural modeling, and functional genetics in zebrafish and Caenorhabditis elegans to identify previously unknown Scm3/HJURP orthologs that localize to centromeres and whose loss causes catastrophic mitotic failure. We further show that Drosophila CAL1, long considered a functional analog, is instead a highly diverged Scm3/HJURP ortholog. Despite rapid primary-sequence divergence, predicted and known structures reveal a broadly conserved CENPA-H4-binding scm3 fold across fungi, vertebrates, nematodes, insects, and basally-branching metazoans. Our work demonstrates how rapid divergence can obscure the broad conservation of essential centromere machinery and provides a broadly applicable strategy to unmasking missing orthologs.

Summary statementAnimals encode a rapidly evolving, essential cell cycle gene previously thought to be absent.
]]></description>
<dc:creator>Hollis, J. A.</dc:creator>
<dc:creator>Stonick, J. A.</dc:creator>
<dc:creator>Topalidou, I.</dc:creator>
<dc:creator>Young, J. M.</dc:creator>
<dc:creator>Moens, C. B.</dc:creator>
<dc:creator>Lehrbach, N. J.</dc:creator>
<dc:creator>Campbell, M. G.</dc:creator>
<dc:creator>Malik, H. S.</dc:creator>
<dc:date>2026-03-06</dc:date>
<dc:identifier>doi:10.64898/2026.03.04.709615</dc:identifier>
<dc:title><![CDATA[Remote homology and functional genetics unmask deeply preserved Scm3/HJURP orthologs in metazoans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.08.708905v1?rss=1">
<title>
<![CDATA[
Hunting for microsatellite instability in long-read data with Owl 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.08.708905v1?rss=1</link>
<description><![CDATA[
Microsatellite instability (MSI) is a key biomarker of mismatch repair deficiency and response to immunotherapy, yet most existing genomic detection methods are optimized for short-read sequencing and rely on small panels of homopolymer markers, limiting the ability to characterize genome-wide and motif-specific patterns of instability. Here we present Owl, a bioinformatic tool for quantifying MSI from long-read (PacBio) genomic data. Owl leverages a genome-wide marker set of more than 140,000 microsatellite repeats ranging from 1-6 bp in length to measure MSI across a phased genome. Using a wrap-around alignment algorithm, Owl constructs repeat-length distributions at each marker site and flags somatic instability using the coefficient of variation. We applied Owl to screen for markers with stable coverage, phasing, and baseline variation across 131 diverse genomes from the Human Pangenome Reference Consortium, where Owl scores ranged from 1.4% to 5.4% of markers exceeding the instability threshold. When applied to 19 cancer cell lines and one diffuse astrocytoma tumor-normal pair, Owl identified five MSI-high genomes with 15-18% unstable markers and showed close concordance with an Illumina DRAGEN MSI assay for the astrocytoma sample. Motif-level analyses revealed shared enrichment of short homopolymer and dinucleotide (A- and AT-rich) repeats across MSI-high cancers, and additionally uncovered a distinct pattern of elevated GGAA microsatellite instability in Ewing sarcoma cell lines, consistent with the known role of the EWS::FLI1 fusion protein at GGAA-rich regulatory elements. Owl is implemented in Rust and integrated into the PacBio HiFi Somatic workflow, providing a scalable framework for MSI analysis from long-read sequencing focused on repeat instability specifically in tumor samples.
]]></description>
<dc:creator>Kronenberg, Z.</dc:creator>
<dc:creator>Chua, K. P.</dc:creator>
<dc:creator>Chaisson, M. J. P.</dc:creator>
<dc:creator>Yoo, B.</dc:creator>
<dc:creator>Lansdon, L.</dc:creator>
<dc:creator>Rowell, W. J.</dc:creator>
<dc:creator>Brandine, G. d. S.</dc:creator>
<dc:creator>Dolzhenko, E.</dc:creator>
<dc:creator>Ikegami, K.</dc:creator>
<dc:creator>Huang, K. K.</dc:creator>
<dc:creator>Tan, P.</dc:creator>
<dc:creator>Bhise, S.</dc:creator>
<dc:creator>Fan, E.</dc:creator>
<dc:creator>Mendoza, M.</dc:creator>
<dc:creator>O'Donnell, E.</dc:creator>
<dc:creator>Pastinen, T.</dc:creator>
<dc:creator>Lawlor, E. R.</dc:creator>
<dc:creator>Furlan, S. N.</dc:creator>
<dc:creator>Farooqi, M. S.</dc:creator>
<dc:creator>Eberle, M. A.</dc:creator>
<dc:date>2026-03-10</dc:date>
<dc:identifier>doi:10.64898/2026.03.08.708905</dc:identifier>
<dc:title><![CDATA[Hunting for microsatellite instability in long-read data with Owl]]></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/10.64898/2026.03.10.710574v1?rss=1">
<title>
<![CDATA[
Therapeutic scheduling of WEE1 inhibition preserves T cell function and promotes immune control of HPV⁺ tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.10.710574v1?rss=1</link>
<description><![CDATA[
Human papillomavirus-associated oropharyngeal squamous cell carcinoma (HPV OPC) is driven by viral E6 and E7 oncoproteins, which disrupt G1 checkpoint control and impose selective dependency on WEE1-mediated G2/M regulation. While this vulnerability confers sensitivity to WEE1 inhibition, its immunologic consequences remain poorly defined, and the challenge of eliciting antitumor immunity without compromising immune fitness has limited clinical translation. Here, we show that WEE1 inhibition elicits durable antitumor immunity in immunocompetent models of HPV OPC. Using murine and human preclinical systems, we demonstrate that the WEE1 inhibitor azenosertib (ZN-c3) mediates tumor control through both cell-autonomous cytotoxicity and immune-dependent mechanisms requiring T cells and conventional dendritic cells. Mechanistically, HPV tumor cells are deficient in STING signaling and fail to mount canonical type I interferon responses. Instead, tumor cell-intrinsic cGAS drives immune activation through STING-competent host cells within the tumor microenvironment, revealing a non-cell-autonomous relay that circumvents viral immune evasion. Intermittent WEE1 inhibition preserves T cell fitness while maintaining antitumor efficacy, and mice achieving complete responses develop immunologic memory capable of rejecting tumor rechallenge. These findings establish intermittent WEE1 inhibition as an immune-permissive therapeutic strategy that enables antigen-specific T cell responses in HPV-driven malignancies and provides a mechanistic rationale for combination with immunotherapy.
]]></description>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Tao, Y.</dc:creator>
<dc:creator>Rahgav, L.</dc:creator>
<dc:creator>Gray-Gaillard, S.</dc:creator>
<dc:creator>Hussaini, Y.</dc:creator>
<dc:creator>Pan, M.</dc:creator>
<dc:creator>Shamber, J.</dc:creator>
<dc:creator>Kwak, J.</dc:creator>
<dc:creator>Park, S. L.</dc:creator>
<dc:creator>Cramer, J.</dc:creator>
<dc:creator>Stoltz, R.</dc:creator>
<dc:creator>Patria, J.</dc:creator>
<dc:creator>Swanger, J.</dc:creator>
<dc:creator>Liu, K.</dc:creator>
<dc:creator>Sannigrahi, M. K.</dc:creator>
<dc:creator>Houghton, M.</dc:creator>
<dc:creator>Rodriguez, C.</dc:creator>
<dc:creator>Carey, R. M.</dc:creator>
<dc:creator>Brody, R.</dc:creator>
<dc:creator>Rajasekaran, K.</dc:creator>
<dc:creator>Weinstein, G.</dc:creator>
<dc:creator>Linette, G. P.</dc:creator>
<dc:creator>Carreno, B. M.</dc:creator>
<dc:creator>Painter, M. M.</dc:creator>
<dc:creator>Wherry, E. J.</dc:creator>
<dc:creator>Clurman, B.</dc:creator>
<dc:creator>Basu, D.</dc:creator>
<dc:creator>Diab, A.</dc:creator>
<dc:date>2026-03-12</dc:date>
<dc:identifier>doi:10.64898/2026.03.10.710574</dc:identifier>
<dc:title><![CDATA[Therapeutic scheduling of WEE1 inhibition preserves T cell function and promotes immune control of HPV⁺ tumors]]></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/10.64898/2026.03.13.711714v1?rss=1">
<title>
<![CDATA[
Proteomics reveals extensive phosphoregulation of outer kinetochore protein KNL1 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.13.711714v1?rss=1</link>
<description><![CDATA[
Microtubules attach to kinetochores to facilitate chromosome movement to opposite spindle poles. Defective kinetochore-microtubule attachments lead to phosphorylation of the outer kinetochore protein KNL1 at conserved MELT motifs, which triggers spindle assembly checkpoint activation and recruitment of the fibrous corona. To identify additional phosphorylation sites that regulate kinetochores, we treated HEK 293T/17 cells with nocodazole, paclitaxel, or STLC to create defective kinetochore-microtubule attachment states. We then purified KNL1 and performed proteomics and identified 111 phosphorylation sites on KNL1, including several that may be attachment-state specific. These data demonstrate that KNL1 is extensively phosphoregulated in response to treatment with microtubule-disrupting compounds.
]]></description>
<dc:creator>Jurasin, A. C.</dc:creator>
<dc:creator>Frank, A. R.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2026-03-15</dc:date>
<dc:identifier>doi:10.64898/2026.03.13.711714</dc:identifier>
<dc:title><![CDATA[Proteomics reveals extensive phosphoregulation of outer kinetochore protein KNL1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.17.712396v1?rss=1">
<title>
<![CDATA[
Estrogen receptor-positive cell line xenograft models recapitulate metastatic dissemination and endocrine response of invasive lobular breast carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.17.712396v1?rss=1</link>
<description><![CDATA[
Invasive lobular breast carcinoma (ILC), the most common special histological subtype of breast cancer, is characterized by nearly universal expression of estrogen receptor alpha (ER) and unique sites of metastases, neither of which is fully recapitulated by genetically engineered mouse models. Using reporter-labeled ILC mouse xenografts, herein we used mammary fat pad, tail vein and intracardiac orthotopic growth to analyze spontaneous and experimental metastasis and gene expression. We observed ER-positive primary tumors with single-file histology and collagen deposition, and spontaneous metastasis from the mammary fat pad to bones, ovaries, and brain including the leptomeninges, thereby closely mirroring the growth and metastatic spread of human ILC. Brain metastases showed strong ER staining, confirmed by sequencing analyses which identified estrogen signaling as top activated pathway, and the lesions exhibited robust response to endocrine therapy. In summary, we report endocrine responsive mammary fat pad, tail vein and intracardiac xenografts that faithfully demonstrate unique ILC features and can serve as invaluable pre-clinical translational platforms for validating candidate ILC genetic drivers and testing novel therapeutics.
]]></description>
<dc:creator>Tasdemir, N.</dc:creator>
<dc:creator>Savariau, L.</dc:creator>
<dc:creator>Scott, J.</dc:creator>
<dc:creator>Latoche, J.</dc:creator>
<dc:creator>Biery, K.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Bossart, E.</dc:creator>
<dc:creator>Sreekumar, S.</dc:creator>
<dc:creator>Brown, D.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Watters, R.</dc:creator>
<dc:creator>Nasrazadani, A.</dc:creator>
<dc:creator>Qin, Y.</dc:creator>
<dc:creator>Cao, Y.</dc:creator>
<dc:creator>Chen, F.</dc:creator>
<dc:creator>Tseng, G.</dc:creator>
<dc:creator>Castro, C.</dc:creator>
<dc:creator>Anderson, C. J.</dc:creator>
<dc:creator>Atkinson, J.</dc:creator>
<dc:creator>Hooda, J.</dc:creator>
<dc:creator>Lucas, P. C.</dc:creator>
<dc:creator>Davidson, N.</dc:creator>
<dc:creator>LEE, A. V.</dc:creator>
<dc:creator>Oesterreich, S.</dc:creator>
<dc:date>2026-03-18</dc:date>
<dc:identifier>doi:10.64898/2026.03.17.712396</dc:identifier>
<dc:title><![CDATA[Estrogen receptor-positive cell line xenograft models recapitulate metastatic dissemination and endocrine response of invasive lobular breast carcinoma]]></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/10.64898/2026.03.17.711896v1?rss=1">
<title>
<![CDATA[
Translating Histopathology Foundation Model Embeddings into Cellular and Molecular Features for Clinical Studies 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.17.711896v1?rss=1</link>
<description><![CDATA[
AI-powered pathology foundation models provide general-purpose representations of histopathological images by encoding image tiles into numerical embeddings. However, these embeddings are not directly interpretable in biological or clinical terms and must be translated into biologically meaningful features, such as cell-type composition or gene expression, to enable downstream clinical applications. To bridge this gap, we developed STpath, a framework that integrates histopathology image embeddings derived from existing pathology foundation models with matched, spatially resolved transcriptomics data. STpath consists of cancer-specific XGBoost models trained to infer cell-type compositions and gene expression from histopathology image tiles. We evaluated STpath in colorectal and breast cancer datasets and showed that it provides accurate estimates of the composition of major cell types and the expression of a subset of genes, with further performance gains achieved by combining embeddings from multiple foundation models. Finally, we demonstrated that STpath inferred features that can be used in downstream studies to evaluate their associations with clinical outcomes.
]]></description>
<dc:creator>Cui, S.</dc:creator>
<dc:creator>Sui, Z.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Matkowskyj, K. A.</dc:creator>
<dc:creator>Yu, M.</dc:creator>
<dc:creator>Grady, W. M.</dc:creator>
<dc:creator>Sun, W.</dc:creator>
<dc:date>2026-03-19</dc:date>
<dc:identifier>doi:10.64898/2026.03.17.711896</dc:identifier>
<dc:title><![CDATA[Translating Histopathology Foundation Model Embeddings into Cellular and Molecular Features for Clinical Studies]]></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/10.64898/2026.03.21.712571v1?rss=1">
<title>
<![CDATA[
Metabolic specialization structures gut bacterial niches and drives colorectal cancer progression 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.21.712571v1?rss=1</link>
<description><![CDATA[
Despite the established association between the gut microbiome and colorectal cancer (CRC), the functional distinction between microbial passengers and drivers of CRC progression remains unresolved. Here, we collected stool, blood, as well as paired tumor, and normal mucosa tissues from seventy-seven CRC patients to characterize the systemic and localized impact of the gut microbiome on early- and late-stage CRC. By deep shotgun metagenomic sequencing, we identified distinct bacterial species and functions residing in tumor versus normal mucosa, highlighting an enrichment of oral-associated bacteria in tumor tissues. Several of these species remained undetected in the stool microbiome analysis. We further combined bacterial culturing with untargeted metabolomics of bacteria enriched in tumor and normal mucosa tissues, revealing distinct clusters of metabolic potential. Functional testing of multiple members from one cluster comprising both tumor- and mucosa-enriched species revealed Leptotrichia wadei as a pro-tumorigenic bacterium in a murine CRC model. Single-nucleus RNA sequencing and in vitro experiments further demonstrated that L. wadei and its secretome induces M2 macrophage polarization to promote tumor growth. Overall, our study shows that metabolic specialization structures microbial colonization niches, while species-specific metabolic outputs identify functional drivers of CRC progression, and uncovers L. wadei as an oncogenic bacterium in CRC.
]]></description>
<dc:creator>Xu, L.-L.</dc:creator>
<dc:creator>Seelbinder, B.</dc:creator>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Kuo, T.-H.</dc:creator>
<dc:creator>Sae-Ong, T.</dc:creator>
<dc:creator>Treibmann, S.</dc:creator>
<dc:creator>Damerell, V.</dc:creator>
<dc:creator>Brobeil, A.</dc:creator>
<dc:creator>Richter, K. M.</dc:creator>
<dc:creator>Mueller, M.</dc:creator>
<dc:creator>Toriola, A. T.</dc:creator>
<dc:creator>Shibata, D.</dc:creator>
<dc:creator>Li, C. I.</dc:creator>
<dc:creator>Byrd, D. A.</dc:creator>
<dc:creator>Figueiredo, J. C.</dc:creator>
<dc:creator>Hardikar, S.</dc:creator>
<dc:creator>Zielinski, C. E.</dc:creator>
<dc:creator>Bleckmann, A.</dc:creator>
<dc:creator>Ni, Y.</dc:creator>
<dc:creator>Correia-Melo, C.</dc:creator>
<dc:creator>Zimmermann, M.</dc:creator>
<dc:creator>Ulrich, C. M.</dc:creator>
<dc:creator>Gigic, B.</dc:creator>
<dc:creator>Panagiotou, G.</dc:creator>
<dc:date>2026-03-23</dc:date>
<dc:identifier>doi:10.64898/2026.03.21.712571</dc:identifier>
<dc:title><![CDATA[Metabolic specialization structures gut bacterial niches and drives colorectal cancer progression]]></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/10.64898/2026.03.23.713714v1?rss=1">
<title>
<![CDATA[
Pluripotency Factors Modulate Interferon Signaling in Embryonic Stem Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.23.713714v1?rss=1</link>
<description><![CDATA[
Despite lacking a robust interferon response, pluripotent stem cells remain highly resistant to viral infection, in part through the constitutive expression of immune genes traditionally classified as interferon-stimulated genes. While interferon signaling has been shown to be incompatible with the maintenance of pluripotency, the molecular mechanisms underlying this relationship remain poorly understood. Here, we investigate the transcriptional response of human embryonic stem cells (hESCs) to infection with a potent activator of the interferon response, an influenza A virus mutant lacking the viral NS1 protein. Single-cell RNA sequencing revealed that while most hESCs remain unresponsive to infection, a distinct subpopulation expresses type I and III interferons. Notably, only interferon-expressing cells mounted a robust antiviral response, characterized by strong induction of interferon-stimulated genes. In contrast to the bulk hESC population, interferon responding cells exhibited reduced expression of core pluripotency factors as well as negative regulators of interferon signaling, such as SOCS1 and SPRY4. Depletion of SOCS1 enabled hESCs to respond robustly to interferon stimulation, showing that this negative regulator is a key suppressor of interferon signaling in pluripotent stem cells. We further show that SOCS1 and additional negative regulators of IFN signaling are intrinsically expressed in hESCs and are transcriptionally controlled by pluripotency factors, such as NANOG, SOX2 and OCT4. Together, our findings support a model in which pluripotency factors regulate intrinsic immune gene expression, including negative regulators of interferon signaling, thereby suppressing canonical interferon signaling to preserve pluripotency while maintaining antiviral resistance.

IMPORTANCEBy combining single-cell transcriptomics with functional studies, we demonstrate that the pluripotency transcriptional program active in pluripotent stem cells coordinately regulates pluripotency factors, antiviral genes, and negative regulators of interferon signaling. This integrated control enables pluripotent stem cells to achieve effective protection against viral infection while preserving their differentiation potential, providing new insights into how innate immunity is selectively constrained in pluripotent stem cells. These findings have important implications for stem cell-based therapies, where transient modulation of antiviral responses without disrupting pluripotency could improve therapeutic efficacy. More broadly, this work advances our understanding of interferon regulation that could inform the development of antiviral strategies that enhance protective immune responses while limiting harmful or unwanted inflammatory signaling.
]]></description>
<dc:creator>Yang, Q.</dc:creator>
<dc:creator>Padilla-Galvez, M.</dc:creator>
<dc:creator>Uhl, S.</dc:creator>
<dc:creator>Eggenberger, J.</dc:creator>
<dc:creator>Kogut, S.</dc:creator>
<dc:creator>Becker, S.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Rosenberg, B. R.</dc:creator>
<dc:creator>Blanco-Melo, D.</dc:creator>
<dc:date>2026-03-25</dc:date>
<dc:identifier>doi:10.64898/2026.03.23.713714</dc:identifier>
<dc:title><![CDATA[Pluripotency Factors Modulate Interferon Signaling in Embryonic Stem Cells]]></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/10.64898/2026.03.23.713807v1?rss=1">
<title>
<![CDATA[
Nextstrain automates real-time phylodynamic analysis of open data for endemic and emerging pathogens 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.23.713807v1?rss=1</link>
<description><![CDATA[
MotivationGenome sequencing provides an exceptional window into the evolutionary and epidemiological dynamics of endemic and emerging pathogens, and thus allows for better, more targeted, public health interventions. Online genomic surveillance platforms can provide near real-time insight into these dynamics.

ResultsNextstrain provides continually updated real-time genomic surveillance for 21 viruses and the bacterial pathogen Mycobacterium tuberculosis, with most analyses relying solely on open sequence data. Each pathogen includes steps to fetch and curate open data, classify sequences using established nomenclature systems, perform phylodynamic analyses, and share the results publicly. These analyses are automated, with most running daily to provide continually updated snapshots of pathogen evolution.

Availability and ImplementationAll source code is available at https://github.com/nextstrain. Phylodynamic results can be visualized and downloaded at https://nextstrain.org/pathogens, and open sequence data and curated metadata are available at https://nextstrain.org/pathogens/files.
]]></description>
<dc:creator>Andrews, K. R.</dc:creator>
<dc:creator>Chang, J.</dc:creator>
<dc:creator>Roemer, C.</dc:creator>
<dc:creator>Hadfield, J.</dc:creator>
<dc:creator>Lin, V.</dc:creator>
<dc:creator>Brito, A. F.</dc:creator>
<dc:creator>Daodu, R.</dc:creator>
<dc:creator>Joia, I. A.</dc:creator>
<dc:creator>Kistler, K.</dc:creator>
<dc:creator>Li, A. W.</dc:creator>
<dc:creator>Moncla, L. H.</dc:creator>
<dc:creator>Paredes, M. I.</dc:creator>
<dc:creator>Kuhnert, D.</dc:creator>
<dc:creator>Torres, L. M.</dc:creator>
<dc:creator>Voitl, L.</dc:creator>
<dc:creator>Aksamentov, I.</dc:creator>
<dc:creator>Hodcroft, E. B.</dc:creator>
<dc:creator>Huddleston, J.</dc:creator>
<dc:creator>McCrone, J. T.</dc:creator>
<dc:creator>Anderson, J. S.</dc:creator>
<dc:creator>Sibley, T. R.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Neher, R. A.</dc:creator>
<dc:creator>Bedford, T.</dc:creator>
<dc:date>2026-03-26</dc:date>
<dc:identifier>doi:10.64898/2026.03.23.713807</dc:identifier>
<dc:title><![CDATA[Nextstrain automates real-time phylodynamic analysis of open data for endemic and emerging pathogens]]></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/10.64898/2026.03.24.713952v1?rss=1">
<title>
<![CDATA[
A codon-sensitive conformational switch gates commitment to translation start sites 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.24.713952v1?rss=1</link>
<description><![CDATA[
Human translation initiation requires single-nucleotide precision to establish the reading frame, yet initiation at non-AUG codons plays key roles in gene expression. How the initiation machinery balances precision with this regulated flexibility remains unclear. Here, we define a conformational branchpoint governed by the human initiation factor eIF5 that gates commitment to start codons. Using single-molecule and structural approaches, we demonstrate that eIF5 reversibly occupies two conformations, which depends on a strictly conserved loop in the protein that monitors start codon identity. AUG codons favor the conformation that is stabilized by an eIF5-stimulated GTP hydrolysis step, which commits the complex to the start site. Non-AUG codons favor a  standby conformation that destabilizes eIF5 and likely overlaps the binding site of an ancient structural homolog. This branchpoint complements enforcement of start codon fidelity by upstream steps and intrinsically controls the efficiency of non-AUG initiation.
]]></description>
<dc:creator>McGuire, S. F.</dc:creator>
<dc:creator>Chan, M. C.</dc:creator>
<dc:creator>Chan, T. C.</dc:creator>
<dc:creator>Pachikara, N.</dc:creator>
<dc:creator>Alleman, E. M.</dc:creator>
<dc:creator>Sikora, V. M.</dc:creator>
<dc:creator>Subramaniam, A. R.</dc:creator>
<dc:creator>Campbell, M. G.</dc:creator>
<dc:creator>Lapointe, C. P.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.24.713952</dc:identifier>
<dc:title><![CDATA[A codon-sensitive conformational switch gates commitment to translation start sites]]></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/10.64898/2026.03.25.714319v1?rss=1">
<title>
<![CDATA[
An interdependent Cbf1-CCAN interaction stabilizes the budding yeast kinetochore 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.25.714319v1?rss=1</link>
<description><![CDATA[
Chromosome segregation requires the proper assembly of kinetochores on centromeric DNA. The kinetochore is a complex multi-protein machine comprising more than 40 distinct proteins, but the functional roles of many components remain unclear. One such protein is the yeast transcription factor Cbf1, which directly binds to budding yeast centromeric DNA. Loss of Cbf1 significantly increases the rate of chromosome missegregation, however its precise molecular mechanism of action is unknown. It was recently found that Cbf1 inhibits transcription through the centromere by preventing the untimely pericentromeric transcriptional readthrough via a roadblock mechanism. Intriguingly, restoring the transcriptional roadblock in the absence of Cbf1 binding only partially rescued chromosome missegregation, indicating that Cbf1 performs additional centromeric activities. Here, we show that Cbf1 promotes inner kinetochore assembly both in vitro and in vivo. This assembly function depends on the direct interaction between Cbf1 and Okp1. Moreover, we found that Cbf1s stable association with the centromere requires its interaction with the inner kinetochore, revealing an interdependent interaction essential for the assembly and stability of the kinetochore. Thus, Cbf1 functions as a centromere-anchored hub that couples transcriptional roadblocking to CCAN assembly and kinetochore stability.
]]></description>
<dc:creator>Hedouin, S.</dc:creator>
<dc:creator>Hu, C.</dc:creator>
<dc:creator>Biggins, S.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.25.714319</dc:identifier>
<dc:title><![CDATA[An interdependent Cbf1-CCAN interaction stabilizes the budding yeast kinetochore]]></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/10.64898/2026.03.26.711478v1?rss=1">
<title>
<![CDATA[
High-dimensional multiomics reveals perturbations to IL-6/IL-6R axis and RUNX3 in CD4+ T cells during third trimester pregnancy 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.26.711478v1?rss=1</link>
<description><![CDATA[
ObjectivesCD4+ T cells play key roles in regulating immune responses during pregnancy, therefore we aimed to understand the CD4+ T cell surface proteome and transcriptome during pregnancy.

MethodsCD4+ T cells were analysed in blood and decidua from term-pregnancies (>37 weeks), and non-pregnant blood. >350 surface proteins were screened via flow cytometry, and transcriptomes were analysed using single-cell RNA sequencing with >130 CITE-seq barcoded antibodies.

ResultsSurface protein screening identified changes to ILT4/CD85d, CD9, IFN-{gamma} receptor {beta}-chain, CX3CR1 and CCR5 in the pregnant blood and decidual CD4+ T cells. CX3CR1 and CCR5 had the highest expression on the effector-memory T cell (TEM) subset in the blood, with expression consistent across subsets in decidua. CD126/IL-6R was lower in pregnant blood and decidual CD4+ T cells, while scRNAseq identified enrichment in the IL-6R signalling pathway in naive CD4+ T cells in pregnant blood. Both sIL-6R and IL-6 concentrations were increased in plasma during pregnancy, suggesting perturbations to the IL-6/IL-6R signalling axis. Meanwhile, decidual CD4+ T cells had increased expression of transcription factor RUNX3 in the CD69+ tissue-resident-like subset.

ConclusionsOur findings demonstrate altered molecular expression in CD4+ T cells during pregnancy. This provides important mechanistic insight of their adaptation and regulation during placental development, which may drive placental dysfunction or pregnancy complications including preeclampsia, fetal growth restriction and stillbirth. These new data may inform future studies that focus on determining the significance of differentially- expressed immune features in pregnancy to identify potential targets for immune modulation to treat pregnancy complications and infections.
]]></description>
<dc:creator>Habel, J.</dc:creator>
<dc:creator>Nguyen, T. H. O.</dc:creator>
<dc:creator>de Alwis, N.</dc:creator>
<dc:creator>Allen, E. K.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Juno, J. A.</dc:creator>
<dc:creator>Kent, S. J.</dc:creator>
<dc:creator>Bond, K.</dc:creator>
<dc:creator>Williamson, D.</dc:creator>
<dc:creator>Lappas, M.</dc:creator>
<dc:creator>Hannan, N.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Schroeder, J.</dc:creator>
<dc:creator>Crawford, J. C.</dc:creator>
<dc:creator>Thomas, P.</dc:creator>
<dc:creator>Kedzierska, K.</dc:creator>
<dc:creator>Rowntree, L.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.711478</dc:identifier>
<dc:title><![CDATA[High-dimensional multiomics reveals perturbations to IL-6/IL-6R axis and RUNX3 in CD4+ T cells during third trimester pregnancy]]></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/10.64898/2026.03.26.714571v1?rss=1">
<title>
<![CDATA[
Cellector: A tool to detect foreign genotype cells in scRNAseq data with applications in leukemia and microchimerism. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.26.714571v1?rss=1</link>
<description><![CDATA[
The existence of rare, genetically distinct cells can occur in various samples such as transplant patients, naturally occurring microchimerism between maternal and fetal tissues, and cancer samples with sufficient mutational burden. Computational methods for detecting these foreign cells are vital to studying these biological conditions. An application that is of particular interest is that of leukemia patients post hematopoietic cell transplant (HCT). In many leukemias, a primary therapy is HCT, after which, the primary genotype of the bone marrow and blood cells should be of donor origin. If cells exist that are of the patients genotype and the cell type lineage of the particular leukemia, this is known as measurable residual disease (MRD). If the MRD is high enough, this may represent a relapse of the patients leukemia. Furthermore, accurately estimating the MRD is important for driving clinical decision making for these patients. Here we present Cellector, a computational method for identifying rare foreign genotype cells in single cell RNAseq (scRNAseq) datasets. We show cellector accurately detects microchimeric cells down to an exceedingly low percentage of these cells present (0.05% or lower).
]]></description>
<dc:creator>Heaton, H.</dc:creator>
<dc:creator>Behboudi, R.</dc:creator>
<dc:creator>Ward, C.</dc:creator>
<dc:creator>Weerakoon, M.</dc:creator>
<dc:creator>Kanaan, S.</dc:creator>
<dc:creator>Reichle, S.</dc:creator>
<dc:creator>Hunter, N.</dc:creator>
<dc:creator>Furlan, S.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.714571</dc:identifier>
<dc:title><![CDATA[Cellector: A tool to detect foreign genotype cells in scRNAseq data with applications in leukemia and microchimerism.]]></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/10.64898/2026.03.27.714269v1?rss=1">
<title>
<![CDATA[
Fc receptor dependent and independent mechanisms of antibody-mediatedenhancement of immune responses 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714269v1?rss=1</link>
<description><![CDATA[
Immune memory responses are rapid and qualitatively distinct from primary responses. They typically develop in the presence of antigen-experienced memory T and B cells and pre-existing antibodies. Although the contribution of T and B cells to recall responses is well defined, the contribution of antibody "memory" and the mechanisms by which pre-existing antibodies modulate the development of germinal center and plasma cell responses is not precisely understood. Here we report on mechanisms that mediate antibody enhancement of germinal center (GC) and plasmablast (PB) compartments, and the parallel process by which they change the affinity threshold for B cell recruitment into immune responses. The data indicate that antibody-mediated enhancement of GC and PB responses is Fc gamma receptor (Fc{gamma}R) dependent and largely complement receptor 1 and 2 (CR1/2) independent. In contrast, the reduction in the affinity threshold for GC entry is independent of both Fc{gamma}Rs and CR1/2.

SummaryCipolla et al. show that antibody can modulate immune responses via both Fc gamma receptor dependent and independent mechanisms. These mechanisms influence both the magnitude and composition of the germinal center response.
]]></description>
<dc:creator>Cipolla, M.</dc:creator>
<dc:creator>MacLean, A. J.</dc:creator>
<dc:creator>Hernandez, B.</dc:creator>
<dc:creator>Silva Santos, G. S.</dc:creator>
<dc:creator>Stamatatos, L.</dc:creator>
<dc:creator>Gazumyan, A.</dc:creator>
<dc:creator>Hartweger, H.</dc:creator>
<dc:creator>Merkenschlager, J.</dc:creator>
<dc:creator>Bournazos, S.</dc:creator>
<dc:creator>Ravetch, J.</dc:creator>
<dc:creator>Nussenzweig, M.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714269</dc:identifier>
<dc:title><![CDATA[Fc receptor dependent and independent mechanisms of antibody-mediatedenhancement of immune responses]]></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/10.64898/2026.03.30.715276v1?rss=1">
<title>
<![CDATA[
IMMREP25: Unseen Peptides 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.30.715276v1?rss=1</link>
<description><![CDATA[
T cell receptors (TCRs) can bind to peptides presented by MHC molecules (pMHC) as a first step to trigger a T cell response. Reliable approaches to predict TCR:pMHC binding would have broad applications in clinical diagnostics, therapeutics, and the fundamental understanding of molecular interactions. IMMREP is a community organized series of prediction contests that asks participants to predict TCR:pMHC binding on unpublished datasets. Previous iterations in 2022 and 2023 showed multiple approaches can predict TCR-pMHC binding with significant accuracy (median AUC_0.1[&ge;]0.7) for peptides where experimental data is available ("seen" peptides). In contrast, models did not outperform random guessing for peptides that have no such data available ("unseen" peptides). Here we report on the results of IMMREP25, which focused solely on unseen peptides in order to evaluate the cutting edge of the field. We received 126 named submissions predicting the specificity of 1,000 TCRs against twenty unseen peptides restricted by one of two MHC molecules (HLA-A*02:01 and HLA-B*40:01). The best performing methods showed a macro-AUC_0.1 of 0.60, significantly better than random, demonstrating significant advances in the field. The top performing methods incorporated structural modeling into their approach, indicating that especially for  unseen peptides, a structural understanding aids in the prediction of TCR:pMHC interactions. The results from this benchmark highlight the significant challenges remaining for TCR:pMHC predictions and will inform future method development.
]]></description>
<dc:creator>Richardson, E.</dc:creator>
<dc:creator>Aarts, Y. J. M.</dc:creator>
<dc:creator>Altin, J. A.</dc:creator>
<dc:creator>Baakman, C. A. B.</dc:creator>
<dc:creator>Bradley, P.</dc:creator>
<dc:creator>Chen, B.</dc:creator>
<dc:creator>Clifford, J.</dc:creator>
<dc:creator>Dhar, M.</dc:creator>
<dc:creator>Diepenbroek, D.</dc:creator>
<dc:creator>Fast, E.</dc:creator>
<dc:creator>Gowthaman, R.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Karnaukhov, V.</dc:creator>
<dc:creator>Marzella, D. F.</dc:creator>
<dc:creator>Meysman, P.</dc:creator>
<dc:creator>Nielsen, M.</dc:creator>
<dc:creator>Nilsson, J. B.</dc:creator>
<dc:creator>Deleuran, S. N.</dc:creator>
<dc:creator>Parizi, F. M.</dc:creator>
<dc:creator>Pelissier, A.</dc:creator>
<dc:creator>Pierce, B. G.</dc:creator>
<dc:creator>Rodriguez Martinez, M.</dc:creator>
<dc:creator>Roran A R, D.</dc:creator>
<dc:creator>Saravanakumar, S.</dc:creator>
<dc:creator>Shao, Y.</dc:creator>
<dc:creator>Smit, N.</dc:creator>
<dc:creator>Van Houcke, M.</dc:creator>
<dc:creator>Visani, G. M.</dc:creator>
<dc:creator>Wan, Y.-T. R.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Woods, L.</dc:creator>
<dc:creator>Wuyts, S.</dc:creator>
<dc:creator>Xiao, C.</dc:creator>
<dc:creator>Xue, L. C.</dc:creator>
<dc:creator>IMMREP25 Participant Consortium,</dc:creator>
<dc:creator>Barton, J.</dc:creator>
<dc:creator>Noakes, M.</dc:creator>
<dc:creator>May, D. H.</dc:creator>
<dc:creator>Peters, B.</dc:creator>
<dc:date>2026-04-01</dc:date>
<dc:identifier>doi:10.64898/2026.03.30.715276</dc:identifier>
<dc:title><![CDATA[IMMREP25: Unseen Peptides]]></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/10.64898/2026.03.31.715024v1?rss=1">
<title>
<![CDATA[
Aurora kinase A enables collective invasion and metastasis by endowing a leader cell phenotype and stabilizing Eplin-mediated cohesion with follower cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.31.715024v1?rss=1</link>
<description><![CDATA[
The metastatic process initiates with collective cell invasion into surrounding tissues and axillary nodes, and subsequent colonization at a distant site. Previously, we found collective invasion is augmented during the G2 cell cycle phase, facilitated through Aurora kinase A (AURKA)-mediated centrosome polarization in the leader cell. Here, we identify cell cycle-associated gene signatures as overrepresented in axilla and liver metastatic sites, with AURKA expression strongly correlated with breast cancer metastasis signatures, and pan-cancer patient survival. Then, we show GFP-AURKA expression endows breast epithelia cells with the ability to form metastatic outgrowths within immune-incompetent chicken embryos. Multi-parametric imaging of wound closure assays reveals phenotypes enabled by, and dependent upon AURKA expression. We discover leader cells express AURKA and acquire front-polarized centrosomes, which differentiates them from other cells in the migrating group. Ectopic expression of GFP-AURKA induces a leader cell phenotype. Conversely, inhibition of AURKA activity alters actin dynamics, promotes turnover of cell contacts, and reduces coordination within migrating groups. Specifically, AURKA interacts with the actin regulator EPLIN, and AURKA inhibition localizes EPLIN to lamellipodia and away from E-cadherin-positive contacts. Inhibiting these necessary roles for AURKA may provide a critical barrier against the metastatic spread of human breast carcinoma cells.
]]></description>
<dc:creator>Zhou, B. P.</dc:creator>
<dc:creator>Chu, T. L. H.</dc:creator>
<dc:creator>Gallant, A. K.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Bhat, T. A.</dc:creator>
<dc:creator>Ghorayeb, R.</dc:creator>
<dc:creator>Gough, C.</dc:creator>
<dc:creator>Garcia, R. E.</dc:creator>
<dc:creator>Pujana, M. A.</dc:creator>
<dc:creator>Lim, C. J.</dc:creator>
<dc:creator>Maxwell, C. A.</dc:creator>
<dc:date>2026-04-02</dc:date>
<dc:identifier>doi:10.64898/2026.03.31.715024</dc:identifier>
<dc:title><![CDATA[Aurora kinase A enables collective invasion and metastasis by endowing a leader cell phenotype and stabilizing Eplin-mediated cohesion with follower cells]]></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/10.64898/2026.04.02.715584v1?rss=1">
<title>
<![CDATA[
Bovine H5N1 influenza viruses have adapted to more efficiently use receptors abundant in cattle 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.02.715584v1?rss=1</link>
<description><![CDATA[
Sustained mammal-to-mammal transmission of high pathogenicity H5N1 avian influenza viruses is reshaping the host range of these pathogens. One of the longest-running mammalian transmission chains involves the B3.13 genotype circulating in U.S. dairy cattle which was detected in early 2024. Genomic analyses revealed selection and rapid fixation of haemagglutinin mutations D104G and V147M. We demonstate, via glycomic profiling, that bovine tissues, including the mammary gland, are enriched in N- and O-linked glycans capped with N-glycolylneuraminic acid (NeuGc), a sialic acid absent in humans and birds, which instead express only N-acetylneuraminic acid (NeuAc). Early cattle H5 viruses poorly recognized NeuGc, but D104G and V147M enabled efficient engagement of both NeuAc- and NeuGc-containing receptors. These mutations enhanced replication in bovine mammary tissue without major attenuation of replication in human lung and primary nasal epithelial cells. NeuGc-driven receptor adaptation therefore promotes viral fitness in cattle while potentially limiting immediate zoonotic risk. Deep mutational scanning further identified alternative haemagglutinin substitutions that confer NeuGc usage and represent surveillance markers for emerging cattle H5 lineages.
]]></description>
<dc:creator>Hassard, J. A.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Dadonaite, B.</dc:creator>
<dc:creator>Pekar, J. E.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Richardson, S. A. S.</dc:creator>
<dc:creator>Pinto, R. M.</dc:creator>
<dc:creator>Ramirez Valdez, K.</dc:creator>
<dc:creator>Lemey, P.</dc:creator>
<dc:creator>Quantrill, J. L.</dc:creator>
<dc:creator>Xue, J.</dc:creator>
<dc:creator>Masonou, T.</dc:creator>
<dc:creator>Case, K.-M.</dc:creator>
<dc:creator>Ajeian, J.</dc:creator>
<dc:creator>Woodall, M. N. J.</dc:creator>
<dc:creator>Ross, R. A.</dc:creator>
<dc:creator>Hudson, N.</dc:creator>
<dc:creator>Zhong, K.</dc:creator>
<dc:creator>Cao, H.</dc:creator>
<dc:creator>Jones, S.</dc:creator>
<dc:creator>Klim, H. J.</dc:creator>
<dc:creator>Wasik, B. R.</dc:creator>
<dc:creator>Dermawan, D. N.</dc:creator>
<dc:creator>Sadeyen, J.-R.</dc:creator>
<dc:creator>Werling, D.</dc:creator>
<dc:creator>Yaffy, D.</dc:creator>
<dc:creator>James, J.</dc:creator>
<dc:creator>Nunez, A.</dc:creator>
<dc:creator>Digard, P.</dc:creator>
<dc:creator>Brown, I. H.</dc:creator>
<dc:creator>Goldhill, D. H.</dc:creator>
<dc:creator>Murcia, P. R.</dc:creator>
<dc:creator>Smith, C. M.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Bloom, J. D.</dc:creator>
<dc:creator>Iqbal, M.</dc:creator>
<dc:creator>Barclay, W. S.</dc:creator>
<dc:creator>Haslam, S. M.</dc:creator>
<dc:creator>Peacock, T. P.</dc:creator>
<dc:date>2026-04-06</dc:date>
<dc:identifier>doi:10.64898/2026.04.02.715584</dc:identifier>
<dc:title><![CDATA[Bovine H5N1 influenza viruses have adapted to more efficiently use receptors abundant in cattle]]></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/10.64898/2026.04.13.717827v1?rss=1">
<title>
<![CDATA[
Multi-omic profiling of human antibody-secreting cells reveals diverse subsets sustain durable humoral immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.04.13.717827v1?rss=1</link>
<description><![CDATA[
Antibody-secreting cells (ASCs) provide humoral immunity that can mediate lifelong protection against pathogens. Current classifications cannot delineate the heterogenous functionalities, tissue residencies, and lifespans of human ASC subsets, impeding clinical translation. We applied multi-omic sequencing, spatial proteomics, and functional assays to discover and characterize human bone marrow (BM) ASC subsets. We identified two peripheral subsets (ASCp) also present in blood and three BM-resident subsets (ASCr), comprising a maturation continuum associated with increased mitochondrial networking, diminished antibody secretion, differential transcription factor motif accessibility, and preferential co-localization in homotypic niches. CD19+9+ASCr and CD19-ASCr exhibited poor recovery years after BM transplantation, indicating a strong dependence on supportive niches. Childhood vaccine antigens were recognized by long-lived ASCr subsets in adults and by immature HLA-DR+ASCp, implying ASCs can differentiate without recent antigen exposure. Our results provide new insights into ASC identity, maturation, and longevity and a generalizable framework for study and manipulation of human ASCs.
]]></description>
<dc:creator>Glass, D. R.</dc:creator>
<dc:creator>Dornisch, E. M.</dc:creator>
<dc:creator>Yin, H.</dc:creator>
<dc:creator>Ludmann, S. A.</dc:creator>
<dc:creator>Samudre, A.</dc:creator>
<dc:creator>Kuhl, S.</dc:creator>
<dc:creator>Malone, J.</dc:creator>
<dc:creator>Chander, A.</dc:creator>
<dc:creator>Kaul, S. N.</dc:creator>
<dc:creator>Phalen, C. G.</dc:creator>
<dc:creator>Parthasarathy, V.</dc:creator>
<dc:creator>Dillon, M. A.</dc:creator>
<dc:creator>Genge, P. C.</dc:creator>
<dc:creator>Stuckey, T. J.</dc:creator>
<dc:creator>Anover-Sombke, S. D.</dc:creator>
<dc:creator>Wittig, P. J.</dc:creator>
<dc:creator>Pebworth, M.-P.</dc:creator>
<dc:creator>He, Z.</dc:creator>
<dc:creator>Henderson, K. E.</dc:creator>
<dc:creator>Ravisankar, P.</dc:creator>
<dc:creator>Hernandez, V.</dc:creator>
<dc:creator>Musgrove, B.</dc:creator>
<dc:creator>Mishra, S.</dc:creator>
<dc:creator>Krishnan, U.</dc:creator>
<dc:creator>Thomson, Z. J.</dc:creator>
<dc:creator>Weiss, M.</dc:creator>
<dc:creator>Estep, N.</dc:creator>
<dc:creator>Graybuck, L. T.</dc:creator>
<dc:creator>Angus-Hill, M. L.</dc:creator>
<dc:creator>Gustafson, C. E.</dc:creator>
<dc:creator>Kopp, M. S.</dc:creator>
<dc:creator>Reading, J.</dc:creator>
<dc:creator>Li, X.-j.</dc:creator>
<dc:creator>Viana, M. P.</dc:creator>
<dc:creator>Bumol, T. F.</dc:creator>
<dc:creator>Goldrath, A. W.</dc:creator>
<dc:creator>Sigvardsson, M.</dc:creator>
<dc:creator>Bendall, S. C.</dc:creator>
<dc:creator>Skene, P. J.</dc:creator>
<dc:creator>Green, D. J.</dc:creator>
<dc:creator>Newell, E. W.</dc:creator>
<dc:creator>Tor</dc:creator>
<dc:date>2026-04-17</dc:date>
<dc:identifier>doi:10.64898/2026.04.13.717827</dc:identifier>
<dc:title><![CDATA[Multi-omic profiling of human antibody-secreting cells reveals diverse subsets sustain durable humoral immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
